Use of suspension growth systems in liquid media in the context of somatic embryogenesis of grapevine (Vitis vinifera L.)
Authorship
A.A.C.
Bachelor of Biology
A.A.C.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
A bibliographic study was carried out on somatic embryogenesis in grapevine (Vitis vinifera L.) using suspension culture systems (liquid media). For this purpose, a historical approach on embryogenesis was carried out, where it was analysed which types of suspension culture have been carried out in grapevine and what advantages they represent with respect to embryogenesis in solid media. By comparing the different works published on this subject, a series of advantages of suspension culture can be extracted, among which the following stand out: the production of a greater quantity of plant material in much less time, more uniform cell growth that facilitates the synchronisation of embryos or greater efficiency in terms of genetic transformation programmes, among others. In addition, the establishment of embryogenic suspension cultures is essential for scaling up using bioreactors, a technology that will provide an improvement in embryogenic processes both now and in the future. Likewise, the future of embryogenesis techniques combined with other sciences or disciplines such as artificial intelligence or BigData that can be applied to automate plant production processes and different genotypes at an industrial level, reducing their cost, will also be developed.
A bibliographic study was carried out on somatic embryogenesis in grapevine (Vitis vinifera L.) using suspension culture systems (liquid media). For this purpose, a historical approach on embryogenesis was carried out, where it was analysed which types of suspension culture have been carried out in grapevine and what advantages they represent with respect to embryogenesis in solid media. By comparing the different works published on this subject, a series of advantages of suspension culture can be extracted, among which the following stand out: the production of a greater quantity of plant material in much less time, more uniform cell growth that facilitates the synchronisation of embryos or greater efficiency in terms of genetic transformation programmes, among others. In addition, the establishment of embryogenic suspension cultures is essential for scaling up using bioreactors, a technology that will provide an improvement in embryogenic processes both now and in the future. Likewise, the future of embryogenesis techniques combined with other sciences or disciplines such as artificial intelligence or BigData that can be applied to automate plant production processes and different genotypes at an industrial level, reducing their cost, will also be developed.
Direction
MARTINEZ TRONCOSO, OSCAR (Tutorships)
MARTINEZ TRONCOSO, OSCAR (Tutorships)
Court
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
Optimization of methodologies in liquid chromatography - tandem mass spectrometry for the determination of hair ethyl glucuronide
Authorship
L.A.A.F.
Double bachelor degree in Chemistry and Biology
L.A.A.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Alcohol is a cause of problems in domestic, legal, workplace and economic spheres, so investigating direct alcohol markers is an important task. In this area, analysis of hair ethyl glucuronide (hEtG) is of particular interest, as this matrix has a large detection window and has gained interest in recent years. LC-MS/MS is now the preferred technique for EtG analysis, with HILIC (hydrophilic interaction liquid chromatography) playing a fundamental role. The current Degree Project has studied the effects of the composition, concentration and pH of the mobile phase, as well as the effect of column temperature, in conjunction with different chromatographic methods. Thus, an HILIC-based chromatographic method for determining hEtG was optimised, achieving a LOQ of 2,64 pg/mg, meeting the Society of Hair Testing (SoHT) requirements. Additionally, the impact of various SPE protocols and cartridges on the analytical signal was evaluated.
Alcohol is a cause of problems in domestic, legal, workplace and economic spheres, so investigating direct alcohol markers is an important task. In this area, analysis of hair ethyl glucuronide (hEtG) is of particular interest, as this matrix has a large detection window and has gained interest in recent years. LC-MS/MS is now the preferred technique for EtG analysis, with HILIC (hydrophilic interaction liquid chromatography) playing a fundamental role. The current Degree Project has studied the effects of the composition, concentration and pH of the mobile phase, as well as the effect of column temperature, in conjunction with different chromatographic methods. Thus, an HILIC-based chromatographic method for determining hEtG was optimised, achieving a LOQ of 2,64 pg/mg, meeting the Society of Hair Testing (SoHT) requirements. Additionally, the impact of various SPE protocols and cartridges on the analytical signal was evaluated.
Direction
MOREDA PIÑEIRO, ANTONIO (Tutorships)
CABARCOS FERNANDEZ, PAMELA (Co-tutorships)
MOREDA PIÑEIRO, ANTONIO (Tutorships)
CABARCOS FERNANDEZ, PAMELA (Co-tutorships)
Court
FERNANDEZ RODRIGUEZ, BERTA (Chairman)
ESTEVEZ VALCARCEL, CARLOS MANUEL (Secretary)
YEBRA BIURRUN, MARIA DEL CARMEN (Member)
FERNANDEZ RODRIGUEZ, BERTA (Chairman)
ESTEVEZ VALCARCEL, CARLOS MANUEL (Secretary)
YEBRA BIURRUN, MARIA DEL CARMEN (Member)
Ethanol toxicokinetics in experimental animal models
Authorship
L.A.A.F.
Double bachelor degree in Chemistry and Biology
L.A.A.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Ethyl alcohol consumption is deeply rooted in Europe and multiple pharmacokinetic model were developed with different characteristics to try to describe its elimination over time, being its application to exprimentation animal models a crucial task in order to translate its results to humans. With this interest, a bibliographic review was conducted using the PubMed and Scopus databases reaching the conclusion that the most commonly used animal model for these pharmacokinetic studies is the rat (Rattus norvegicus). Also, factors as the administered dose, the age, sex or pregancy were found to have a significant influency on ethyl alcohol elimination, without clear conclussions about the possible influence of estral cycle. It is not possible to confirm the impact of race on the elimination pharmacokinetic in combination with the before mentioned factors, as well as the pressence of other substances, such as tobacco or cocaine, in combination with ethyl alcohol.
Ethyl alcohol consumption is deeply rooted in Europe and multiple pharmacokinetic model were developed with different characteristics to try to describe its elimination over time, being its application to exprimentation animal models a crucial task in order to translate its results to humans. With this interest, a bibliographic review was conducted using the PubMed and Scopus databases reaching the conclusion that the most commonly used animal model for these pharmacokinetic studies is the rat (Rattus norvegicus). Also, factors as the administered dose, the age, sex or pregancy were found to have a significant influency on ethyl alcohol elimination, without clear conclussions about the possible influence of estral cycle. It is not possible to confirm the impact of race on the elimination pharmacokinetic in combination with the before mentioned factors, as well as the pressence of other substances, such as tobacco or cocaine, in combination with ethyl alcohol.
Direction
POMBO RAMOS, CELIA MARIA (Tutorships)
BERMEJO BARRERA, ANA MARIA (Co-tutorships)
POMBO RAMOS, CELIA MARIA (Tutorships)
BERMEJO BARRERA, ANA MARIA (Co-tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Construction of expression vectors for fluorescent proteins with specific subcellular localization.
Authorship
I.A.P.
Biotechnology Degree (2nd Ed. )
I.A.P.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
The visualization of subcellular dynamics is essential for numerous studies in cell biology. One of the most commonly used approaches is the expression of fluorescent proteins such as enhanced green fluorescent protein (eGFP), enhanced yellow fluorescent protein (eYFP), Cerulean, or the red fluorescent protein from Discosoma sp. (DsRed), which allow tracking intracellular distribution in real time through fluorescence microscopy. To target these proteins to specific compartments, localization signals such as mitochondrial targeting sequences (MTS), nuclear export signals (NES), or nuclear localization signals (NLS) are used. This work aimed to construct a library of expression vectors in Saccharomyces cerevisiae for fluorescent proteins with defined subcellular localization, as well as to validate their expression using fluorescence microscopy. Gateway cloning techniques, vector purification, bacterial and yeast transformations, and microscopy analysis were employed. The results indicate that, among the proteins analyzed, eGFP showed the clearest signal. Cerulean and DsRed displayed signals that were difficult to distinguish from autofluorescence, making them indistinguishable from the cellular background. Regarding localization signals, eGFP showed expression consistent with mitochondrial targeting and nuclear export signals, while the nuclear localization sequence underwent a mutation that prevented its analysis. The development of a functional library of fluorescent vectors will enable studies on colocalization and subcellular dynamics in yeast, contributing to the functional analysis of proteins and the understanding of complex cellular processes.
The visualization of subcellular dynamics is essential for numerous studies in cell biology. One of the most commonly used approaches is the expression of fluorescent proteins such as enhanced green fluorescent protein (eGFP), enhanced yellow fluorescent protein (eYFP), Cerulean, or the red fluorescent protein from Discosoma sp. (DsRed), which allow tracking intracellular distribution in real time through fluorescence microscopy. To target these proteins to specific compartments, localization signals such as mitochondrial targeting sequences (MTS), nuclear export signals (NES), or nuclear localization signals (NLS) are used. This work aimed to construct a library of expression vectors in Saccharomyces cerevisiae for fluorescent proteins with defined subcellular localization, as well as to validate their expression using fluorescence microscopy. Gateway cloning techniques, vector purification, bacterial and yeast transformations, and microscopy analysis were employed. The results indicate that, among the proteins analyzed, eGFP showed the clearest signal. Cerulean and DsRed displayed signals that were difficult to distinguish from autofluorescence, making them indistinguishable from the cellular background. Regarding localization signals, eGFP showed expression consistent with mitochondrial targeting and nuclear export signals, while the nuclear localization sequence underwent a mutation that prevented its analysis. The development of a functional library of fluorescent vectors will enable studies on colocalization and subcellular dynamics in yeast, contributing to the functional analysis of proteins and the understanding of complex cellular processes.
Direction
González Blanco, Miguel (Tutorships)
González Blanco, Miguel (Tutorships)
Court
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
Use of green waste compost for the recovery of compacted urban soils
Authorship
I.B.R.
Bachelor of Biology
I.B.R.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
This study aims to analyze the comprehensive impact of green waste compost application on the properties of compacted urban soil. Results were compared with uncompacted reference soil, untreated compacted soil, and treated imported soil, and analyzed using the Cornell University Comprehensive Soil Health Assessment approach. Compost application significantly increased active carbon, total N and C, available phosphorus, cation exchange capacity (CEC), aggregate stability (AS), percent organic matter (ME Lo), and total protein values. It also demonstrated a positive effect on micronutrient content, respiration, pH, electrical conductivity (EC), exchangeable cations, and water properties. The study demonstrates that the application of green waste compost can improve the health of compacted urban soils.
This study aims to analyze the comprehensive impact of green waste compost application on the properties of compacted urban soil. Results were compared with uncompacted reference soil, untreated compacted soil, and treated imported soil, and analyzed using the Cornell University Comprehensive Soil Health Assessment approach. Compost application significantly increased active carbon, total N and C, available phosphorus, cation exchange capacity (CEC), aggregate stability (AS), percent organic matter (ME Lo), and total protein values. It also demonstrated a positive effect on micronutrient content, respiration, pH, electrical conductivity (EC), exchangeable cations, and water properties. The study demonstrates that the application of green waste compost can improve the health of compacted urban soils.
Direction
PARADELO NUÑEZ, REMIGIO (Tutorships)
PARADELO NUÑEZ, REMIGIO (Tutorships)
Court
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
PET hydrolase production in a bioreactor
Authorship
N.B.D.L.P.C.
Biotechnology Degree (2nd Ed. )
N.B.D.L.P.C.
Biotechnology Degree (2nd Ed. )
Defense date
07.16.2025 10:30
07.16.2025 10:30
Summary
Plastics are one of the main sources of environmental pollution worldwide. In particular, polyethylene terephthalate (PET) is one of the most widely used polymers in sectors such as the textile and food industries. Conventional PET recycling methods, such as mechanical and chemical recycling, involve high energy consumption and the emission of greenhouse gases, which represents an environmental threat. Against this backdrop, enzymatic recycling has emerged as a sustainable alternative, based on the use of hydrolases capable of degrading PET. In this context, this work aims to optimise the production conditions of two PET hydrolases (duraPETase and LCC-ICCG) in Escherichia coli BL21 (DE3), using a 1.75 L culture bioreactor. For this purpose, different post-induction temperatures (37 and 18 degrees Celsius) and expression times (overnight (o/n) vs 4 h) were evaluated. The results indicate that the most efficient condition is to maintain a constant temperature of 37 degrees Celsius after induction, with a duration of expression of 4 h. Yields of 1.677 mg/g biomass for LCC-ICCG and 0.7458 mg/g biomass for duraPETase were obtained under these conditions, which is at least an 11-fold and 4-fold improvement, respectively, compared to the o/n conditions. Although enzyme activity was higher in o/n conditions, both hydrolases retained their activity after 4 h of expression. The viability of E. coli was also assessed during duraPETase expression. Although a significant decrease in bacterial population was observed at the highest production condition (37 degrees Celsius and 4 h post-induction), no toxicity associated with the recombinant protein was detected at prolonged times. This suggests that the decrease in protein concentration observed in o/n expressions is not due to toxic effects, but possibly to the activation of cellular stress response mechanisms that induce the expression of intracellular proteases capable of degrading the recombinant protein.
Plastics are one of the main sources of environmental pollution worldwide. In particular, polyethylene terephthalate (PET) is one of the most widely used polymers in sectors such as the textile and food industries. Conventional PET recycling methods, such as mechanical and chemical recycling, involve high energy consumption and the emission of greenhouse gases, which represents an environmental threat. Against this backdrop, enzymatic recycling has emerged as a sustainable alternative, based on the use of hydrolases capable of degrading PET. In this context, this work aims to optimise the production conditions of two PET hydrolases (duraPETase and LCC-ICCG) in Escherichia coli BL21 (DE3), using a 1.75 L culture bioreactor. For this purpose, different post-induction temperatures (37 and 18 degrees Celsius) and expression times (overnight (o/n) vs 4 h) were evaluated. The results indicate that the most efficient condition is to maintain a constant temperature of 37 degrees Celsius after induction, with a duration of expression of 4 h. Yields of 1.677 mg/g biomass for LCC-ICCG and 0.7458 mg/g biomass for duraPETase were obtained under these conditions, which is at least an 11-fold and 4-fold improvement, respectively, compared to the o/n conditions. Although enzyme activity was higher in o/n conditions, both hydrolases retained their activity after 4 h of expression. The viability of E. coli was also assessed during duraPETase expression. Although a significant decrease in bacterial population was observed at the highest production condition (37 degrees Celsius and 4 h post-induction), no toxicity associated with the recombinant protein was detected at prolonged times. This suggests that the decrease in protein concentration observed in o/n expressions is not due to toxic effects, but possibly to the activation of cellular stress response mechanisms that induce the expression of intracellular proteases capable of degrading the recombinant protein.
Direction
EIBES GONZALEZ, GEMMA MARIA (Tutorships)
BALBOA MENDEZ, SABELA (Co-tutorships)
EIBES GONZALEZ, GEMMA MARIA (Tutorships)
BALBOA MENDEZ, SABELA (Co-tutorships)
Court
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
Caracterization of a new murine model for inducible genetic editing of glial cells
Authorship
E.M.B.V.
Bachelor of Biology
E.M.B.V.
Bachelor of Biology
Defense date
07.16.2025 09:30
07.16.2025 09:30
Summary
In biomedical research, inducible Cre-recombinase models are a precise genetic tool that allows spatial and temporal control of genetic modifications. In the field of astrocyte research and their brain functions, there are inducible Cre-recombinase models that use genes sucha as human glial fibrillary acidic protein, glutamate transporter, or aldehyde dehydrogenase 1-L1 as promoter sequences. All of these are astrocytic markers that restrict the location of the recombinase to these glial cells. Nevertheless, the classic astrocytic marker to define mouse astrocytes is the murine glial fibrillary acidic protein, for wich, until now, there was no inducible Cre-recombinase model using it as a promoter. In this work, I will perform a histological characterization of a new genetically modified mouse line, generated by CRISPR/Cas9 technology, that uses murine glial fibrillary acidic protein, as a promoter: the GFAP-CreERT2. To achive this, I will work alongside the Neuroendocrine Regulation of Metabolism (NeuRoMet) and NeurObesity groups at CiMUS, conducting in vivo experiments, inmunohistochemistry staining techniques, and molecular biology approaches, wich will allow me to confirm the exclusive presence of this inducible Cre-recombinase Model, in astrocytes.
In biomedical research, inducible Cre-recombinase models are a precise genetic tool that allows spatial and temporal control of genetic modifications. In the field of astrocyte research and their brain functions, there are inducible Cre-recombinase models that use genes sucha as human glial fibrillary acidic protein, glutamate transporter, or aldehyde dehydrogenase 1-L1 as promoter sequences. All of these are astrocytic markers that restrict the location of the recombinase to these glial cells. Nevertheless, the classic astrocytic marker to define mouse astrocytes is the murine glial fibrillary acidic protein, for wich, until now, there was no inducible Cre-recombinase model using it as a promoter. In this work, I will perform a histological characterization of a new genetically modified mouse line, generated by CRISPR/Cas9 technology, that uses murine glial fibrillary acidic protein, as a promoter: the GFAP-CreERT2. To achive this, I will work alongside the Neuroendocrine Regulation of Metabolism (NeuRoMet) and NeurObesity groups at CiMUS, conducting in vivo experiments, inmunohistochemistry staining techniques, and molecular biology approaches, wich will allow me to confirm the exclusive presence of this inducible Cre-recombinase Model, in astrocytes.
Direction
GONZALEZ GARCIA, ISMAEL (Tutorships)
LOPEZ PEREZ, MIGUEL ANTONIO (Co-tutorships)
GONZALEZ GARCIA, ISMAEL (Tutorships)
LOPEZ PEREZ, MIGUEL ANTONIO (Co-tutorships)
Court
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
Molecular and biological function of AP-1 transcription factors in Schwann cell demyelination and peripheral nerve repair.
Authorship
S.B.B.
Biotechnology Degree (2nd Ed. )
S.B.B.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 09:00
07.17.2025 09:00
Summary
Schwann cells, responsible for forming myelin sheaths in the peripheral nervous system, have attracted considerable scientific interest due to their key role in axonal regeneration after traumatic injury, adopting a repair phenotype. However, the mechanisms regulating this reprogramming are not yet fully understood. c-Jun, a transcription factor of the AP-1 family, has been identified as a central regulator of this process. Previous research from our laboratory identified other family members, such as Fra1 and Fra2, which also show significant increases in RNA levels after injury. Additionally, earlier studies reported that FRA1 silencing impairs axonal regeneration. These findings suggest that FRA1 and FRA2, alongside c-Jun, could be part of a broader group of AP-1 transcription factors acting as master regulators. This study aims to elucidate the biological and molecular roles of FRA1 and FRA2 by characterizing Schwann cell responses to sciatic nerve injuries in murine models with deletion of both genes. To achieve this, qPCR and Western blot analyses were performed, revealing significant changes in the expression of Olig1 and Ngf and in ATF3 protein levels between wild-type and knockout mice, while myelin protein clearance remained unaffected. At the physiological level, sciatic nerve integrity during regeneration was assessed by electron microscopy imaging, showing no differences between groups. Preliminary immunofluorescence assays were also conducted, though a larger sample size is needed to confirm these findings. These results suggest that FRA1 and FRA2 play a specific role in axonal regeneration, although the observed effect was less pronounced than expected. Further studies with longer follow-up periods are essential to determine whether these proteins influence full recovery after nerve injury.
Schwann cells, responsible for forming myelin sheaths in the peripheral nervous system, have attracted considerable scientific interest due to their key role in axonal regeneration after traumatic injury, adopting a repair phenotype. However, the mechanisms regulating this reprogramming are not yet fully understood. c-Jun, a transcription factor of the AP-1 family, has been identified as a central regulator of this process. Previous research from our laboratory identified other family members, such as Fra1 and Fra2, which also show significant increases in RNA levels after injury. Additionally, earlier studies reported that FRA1 silencing impairs axonal regeneration. These findings suggest that FRA1 and FRA2, alongside c-Jun, could be part of a broader group of AP-1 transcription factors acting as master regulators. This study aims to elucidate the biological and molecular roles of FRA1 and FRA2 by characterizing Schwann cell responses to sciatic nerve injuries in murine models with deletion of both genes. To achieve this, qPCR and Western blot analyses were performed, revealing significant changes in the expression of Olig1 and Ngf and in ATF3 protein levels between wild-type and knockout mice, while myelin protein clearance remained unaffected. At the physiological level, sciatic nerve integrity during regeneration was assessed by electron microscopy imaging, showing no differences between groups. Preliminary immunofluorescence assays were also conducted, though a larger sample size is needed to confirm these findings. These results suggest that FRA1 and FRA2 play a specific role in axonal regeneration, although the observed effect was less pronounced than expected. Further studies with longer follow-up periods are essential to determine whether these proteins influence full recovery after nerve injury.
Direction
Woodhoo , Ashwin (Tutorships)
AYUSO GARCIA, PAULA (Co-tutorships)
Woodhoo , Ashwin (Tutorships)
AYUSO GARCIA, PAULA (Co-tutorships)
Court
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Development of functionalised biomimetic nanocapsules for the study of internalisation and intracellular routes of nanoparticles
Authorship
D.C.N.
Biotechnology Degree (2nd Ed. )
D.C.N.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
The world of nanotechnology is a developing sector of great interest due to the great advances it can bring to many different fields. In the nanobiotechnology sector, nanoparticles stand out as vectors for delivering drugs in various therapies. For this project, the focus is on biomimetic nanosystems as vectors with unique properties and especially useful in delivering their cargo to cells. Thus, biomimetic nanovesicles based on the cell membrane of A549 cells, referred to as cellsomes, were developed, and gold nanoparticles with different degrees of ubiquitination were encapsulated in them in order to modify the cellular internalisation of the nanoparticles and to study their intracellular localisation. The use of electron microscopy, spectroscopy and nanoparticle tracking analysis techniques allowed us to characterise the cellsomes obtained, while the use of flow cytometry and confocal microscopy allowed us to study the internalisation of the nanoparticles and cellsomes as well as their localisation inside the cells. The results obtained show that synthesis and characterisation are feasible and successful, and that gold nanoparticles manage to enter both cellsomes and cells. Moreover, ubiquitination seems to help in the uptake of the nanoparticles, and the encapsulation of the nanoparticles in the cellsomes is able to change their intracellular localisation.
The world of nanotechnology is a developing sector of great interest due to the great advances it can bring to many different fields. In the nanobiotechnology sector, nanoparticles stand out as vectors for delivering drugs in various therapies. For this project, the focus is on biomimetic nanosystems as vectors with unique properties and especially useful in delivering their cargo to cells. Thus, biomimetic nanovesicles based on the cell membrane of A549 cells, referred to as cellsomes, were developed, and gold nanoparticles with different degrees of ubiquitination were encapsulated in them in order to modify the cellular internalisation of the nanoparticles and to study their intracellular localisation. The use of electron microscopy, spectroscopy and nanoparticle tracking analysis techniques allowed us to characterise the cellsomes obtained, while the use of flow cytometry and confocal microscopy allowed us to study the internalisation of the nanoparticles and cellsomes as well as their localisation inside the cells. The results obtained show that synthesis and characterisation are feasible and successful, and that gold nanoparticles manage to enter both cellsomes and cells. Moreover, ubiquitination seems to help in the uptake of the nanoparticles, and the encapsulation of the nanoparticles in the cellsomes is able to change their intracellular localisation.
Direction
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Tutorships)
PELAZ GARCIA, BEATRIZ (Co-tutorships)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Tutorships)
PELAZ GARCIA, BEATRIZ (Co-tutorships)
Court
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
Clinical laboratory contribution to the SERGAS' integrated assistencial process diabetes mellitus type 2
Authorship
I.C.F.
Double bachelor degree in Chemistry and Biology
I.C.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Precocious detection of DM in asymptomatic people can prevent or delay diabetes complications, with previous evidence proving that most of them are related to the duration and severity of hyperglicemia. The SERGAS IAP for DM2 has the objective of orienting the labor of medical professionals in the diagnosis, treatment and control of people with DM2. In this work the contributions of the Clinical Analysis Laboratory to the SERGAS’ IAP for DM2 are evaluated. Following the criteria established by the ADA, the laboratory has established an opportunistic screening for DM2, that consists on adding the determination of HbA1c to those patients without previous diagnosis of DM and an altered basal glycemia, detected in an analytic performed during the years 2023 and 2024. The data extracted from the laboratory’s informatic system, as well as the revision of the clinical history of the patients is summarized in this work. 22571 pacients with altered basal glycemia were detected. 30% of them had HbA1c values in the normal range, while 49% exhibited HbA1c values in the range of prediabetes, and 21% exhibited HbA1c values in the range of diabetes. In this last group, 2310 had also glucose values larger than 126 mg/dl in the same analytic, allowing to establish a one step diabetes diagnosis. 64% of the diabetic pacients went to subsequent analytics with HbA1c solicited in a time interval of 9,1 (std. dev. 5,4) months. The screening allows to identify undiagnosed diabetic pacients in only one analytic, as well as identifying those with unadequatelly controlled. Moreover, it helps put prediabetic pacients in the spotlight, pacients with high risk of developing diabetes, and it also gives the opportunity to act early over diabetes complications.
Precocious detection of DM in asymptomatic people can prevent or delay diabetes complications, with previous evidence proving that most of them are related to the duration and severity of hyperglicemia. The SERGAS IAP for DM2 has the objective of orienting the labor of medical professionals in the diagnosis, treatment and control of people with DM2. In this work the contributions of the Clinical Analysis Laboratory to the SERGAS’ IAP for DM2 are evaluated. Following the criteria established by the ADA, the laboratory has established an opportunistic screening for DM2, that consists on adding the determination of HbA1c to those patients without previous diagnosis of DM and an altered basal glycemia, detected in an analytic performed during the years 2023 and 2024. The data extracted from the laboratory’s informatic system, as well as the revision of the clinical history of the patients is summarized in this work. 22571 pacients with altered basal glycemia were detected. 30% of them had HbA1c values in the normal range, while 49% exhibited HbA1c values in the range of prediabetes, and 21% exhibited HbA1c values in the range of diabetes. In this last group, 2310 had also glucose values larger than 126 mg/dl in the same analytic, allowing to establish a one step diabetes diagnosis. 64% of the diabetic pacients went to subsequent analytics with HbA1c solicited in a time interval of 9,1 (std. dev. 5,4) months. The screening allows to identify undiagnosed diabetic pacients in only one analytic, as well as identifying those with unadequatelly controlled. Moreover, it helps put prediabetic pacients in the spotlight, pacients with high risk of developing diabetes, and it also gives the opportunity to act early over diabetes complications.
Direction
Rodriguez Garcia, Javier (Tutorships)
Riveiro Cruz, Manuel Alberto (Co-tutorships)
Rodriguez Garcia, Javier (Tutorships)
Riveiro Cruz, Manuel Alberto (Co-tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Evaluation of an automatic ion exchange (HPLC) analyzer for the meassurement of glycated hemoglobin
Authorship
I.C.F.
Double bachelor degree in Chemistry and Biology
I.C.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Verifying the analytical parameters of the different equipment present in a clinical laboratory is a crucial step towards giving quality results. Specifically, glycated hemoglobin (HbA1c) is a very important parameter for both diagnosis and control of diabetic patients. This study aims to verify an HbA1c HPLC analyzer, the ADAMS 8190V, that was recently incorporated into the clinical analysis laboratory, with the purpose of verifying its analytical parameters. Clinical and Laboratory Standards Institute (CLSI) guides were used in conjunction with instructions provided by the Sociedad Española de Química Clínica (SEQC) to study imprecision, veracity and linearity. The effect of relevant interferences was also studied. Finally, a comparative study between the ADAMS 8190V and previously certificated HPLC instruments was performed, to verify that the new instruments meet the quality standards required for the technique. The ADAMS 8190V meets the required quality standards and doesn’t show interferences when exposed to most of the typical interferences that may affect its measuring capabilities, nor in the presence of hemoglobin variants. It does show interferences in the presence of high fractions of carbamylated hemoglobin. The ADAMS 8190V has good quality parameters, which coupled with its smaller analysis times, makes it a positive addition to the routine of the clinical analysis laboratory.
Verifying the analytical parameters of the different equipment present in a clinical laboratory is a crucial step towards giving quality results. Specifically, glycated hemoglobin (HbA1c) is a very important parameter for both diagnosis and control of diabetic patients. This study aims to verify an HbA1c HPLC analyzer, the ADAMS 8190V, that was recently incorporated into the clinical analysis laboratory, with the purpose of verifying its analytical parameters. Clinical and Laboratory Standards Institute (CLSI) guides were used in conjunction with instructions provided by the Sociedad Española de Química Clínica (SEQC) to study imprecision, veracity and linearity. The effect of relevant interferences was also studied. Finally, a comparative study between the ADAMS 8190V and previously certificated HPLC instruments was performed, to verify that the new instruments meet the quality standards required for the technique. The ADAMS 8190V meets the required quality standards and doesn’t show interferences when exposed to most of the typical interferences that may affect its measuring capabilities, nor in the presence of hemoglobin variants. It does show interferences in the presence of high fractions of carbamylated hemoglobin. The ADAMS 8190V has good quality parameters, which coupled with its smaller analysis times, makes it a positive addition to the routine of the clinical analysis laboratory.
Direction
Rodriguez Garcia, Javier (Tutorships)
Ortola Devesa, Juan Bautista (Co-tutorships)
Rodriguez Garcia, Javier (Tutorships)
Ortola Devesa, Juan Bautista (Co-tutorships)
Court
Estévez Cabanas, Juan Carlos (Chairman)
GARCIA SUAREZ, LUIS ALBERTO (Secretary)
GONZALEZ NOYA, ANA MARIA (Member)
Estévez Cabanas, Juan Carlos (Chairman)
GARCIA SUAREZ, LUIS ALBERTO (Secretary)
GONZALEZ NOYA, ANA MARIA (Member)
Characterization of COL4A3-COL4A5 Genes in a Cohort of Patients with Glomerular Disease
Authorship
L.C.V.
Biotechnology Degree (2nd Ed. )
L.C.V.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 12:30
07.17.2025 12:30
Summary
Hereditary kidney diseases are present in a very small percentage of the population. Specifically, this project focuses on the study of glomerular diseases, which develop due to defects in the glomerular basement membrane and involve alterations in type IV collagen genes. The COL4A3-COL4A5 genes can present a wide range of variant, some of which are non-pathogenic, while many others are involved in the development of conditions such as Alport syndrome. Genetic testingallows for the early diagnosis of these conditions, both in patients with severe symptoms and in those with milder symptoms or even asymptomatic individuals. The ongoing advancement and refinement of these diagnostic techniques enable more precise analyses of such diseases, the development of targeted therapies, and ultimately the improvement of patients quality of life and overall well-being. Therefore, this study aims to carry out a comprehensive search and analysis of the different pathogenic variants that may be involved in the development of these nephropathies, with the goal of providing accurate and high-quality diagnoses for the future.
Hereditary kidney diseases are present in a very small percentage of the population. Specifically, this project focuses on the study of glomerular diseases, which develop due to defects in the glomerular basement membrane and involve alterations in type IV collagen genes. The COL4A3-COL4A5 genes can present a wide range of variant, some of which are non-pathogenic, while many others are involved in the development of conditions such as Alport syndrome. Genetic testingallows for the early diagnosis of these conditions, both in patients with severe symptoms and in those with milder symptoms or even asymptomatic individuals. The ongoing advancement and refinement of these diagnostic techniques enable more precise analyses of such diseases, the development of targeted therapies, and ultimately the improvement of patients quality of life and overall well-being. Therefore, this study aims to carry out a comprehensive search and analysis of the different pathogenic variants that may be involved in the development of these nephropathies, with the goal of providing accurate and high-quality diagnoses for the future.
Direction
CANDAL SUAREZ, EVA MARIA (Tutorships)
GARCIA MURIAS, MARIA (Co-tutorships)
CANDAL SUAREZ, EVA MARIA (Tutorships)
GARCIA MURIAS, MARIA (Co-tutorships)
Court
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
p107 and its role in the progression of MASLD to MASH in a diet-induced steatohepatitis mouse model.
Authorship
G.A.C.S.
Biotechnology Degree (2nd Ed. )
G.A.C.S.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 09:00
07.17.2025 09:00
Summary
Metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the most common manifestations of metabolic syndrome, a group of disorders derived from obesity, hypertension, and insulin resistance. This disease can progress to more severe stages, such as metabolic dysfunction-associated steatohepatitis (MASH). Recent studies from our laboratory have shown that the absence of p107 improves hepatic metabolism; however, its role in the progression from MASLD to more advanced stages, such as MASH and fibrosis, has not been described. Therefore, the objective of this Thesis is to analyse whether the global absence of p107 has an effect on the progression from MASLD to MASH in a diet-induced steatohepatitis mouse model. To investigate this, histological and molecular parameters related to lipid metabolism and hepatic fibrosis were evaluated. The results obtained suggest that the global absence of p107 is associated with a reduction in hepatic lipid accumulation but could favour fibrosis progression. These findings indicate that p107 may play an important role in the advancement of liver disease to more severe stages, opening up new lines of research to better understand the mechanisms involved.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the most common manifestations of metabolic syndrome, a group of disorders derived from obesity, hypertension, and insulin resistance. This disease can progress to more severe stages, such as metabolic dysfunction-associated steatohepatitis (MASH). Recent studies from our laboratory have shown that the absence of p107 improves hepatic metabolism; however, its role in the progression from MASLD to more advanced stages, such as MASH and fibrosis, has not been described. Therefore, the objective of this Thesis is to analyse whether the global absence of p107 has an effect on the progression from MASLD to MASH in a diet-induced steatohepatitis mouse model. To investigate this, histological and molecular parameters related to lipid metabolism and hepatic fibrosis were evaluated. The results obtained suggest that the global absence of p107 is associated with a reduction in hepatic lipid accumulation but could favour fibrosis progression. These findings indicate that p107 may play an important role in the advancement of liver disease to more severe stages, opening up new lines of research to better understand the mechanisms involved.
Direction
TOVAR CARRO, SULAY AMPARO (Tutorships)
TOVAR CARRO, SULAY AMPARO (Tutorships)
Court
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Biochemical, Molecular and Functional Characterization of a 3D Model Derived from Epicardial Mesenchymal Cells
Authorship
M.C.T.
Bachelor of Biology
M.C.T.
Bachelor of Biology
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Metabolic alterations derived from obesity contribute decisively to the development of cardiovascular diseases. In this context, epicardial adipose tissue stands out for its active role in cardiac dysfunction through the secretion of paracrine, endocrine, and vasocrine signals that affect both the myocardium and the coronary vasculature. Both epicardial and subcutaneous adipose tissues contain mesenchymal cells with a marked potential to induce adipogenesis and angiogenesis processes involved in cardiac remodeling and disease progression. Based on the hypothesis that a three-dimensional (3D) system would better reproduce the interaction between these processes, a 3D cell culture model was designed and characterized using mesenchymal cells isolated from subcutaneous and epicardial adipose tissue of patients undergoing cardiac surgery. Different extracellular matrices were evaluated, and specific treatments were applied to induce differentiation processes. The morphological and functional characterization of the model was carried out using optical microscopy, immunofluorescence, gene expression analysis by real-time PCR, and protein analysis by Western blot. The results confirmed the model’s capacity to reproduce adipogenesis and angiogenesis processes in an integrated manner, evidencing differences depending on the tissue origin. Subcutaneous adipose tissue showed a greater adipogenic capacity and three-dimensional organization, while epicardial adipose tissue exhibited a more inflammatory profile and a less cohesive structure. The angiogenic response was modest in both cases. Furthermore, structural and functional differences were observed between matrices, with Matrigel standing out as the most effective in forming stable spheroids. This model represents a useful preclinical tool to study the interaction between adipogenesis and angiogenesis processes in different tissues, as well as to explore new therapeutic approaches for cardiovascular diseases.
Metabolic alterations derived from obesity contribute decisively to the development of cardiovascular diseases. In this context, epicardial adipose tissue stands out for its active role in cardiac dysfunction through the secretion of paracrine, endocrine, and vasocrine signals that affect both the myocardium and the coronary vasculature. Both epicardial and subcutaneous adipose tissues contain mesenchymal cells with a marked potential to induce adipogenesis and angiogenesis processes involved in cardiac remodeling and disease progression. Based on the hypothesis that a three-dimensional (3D) system would better reproduce the interaction between these processes, a 3D cell culture model was designed and characterized using mesenchymal cells isolated from subcutaneous and epicardial adipose tissue of patients undergoing cardiac surgery. Different extracellular matrices were evaluated, and specific treatments were applied to induce differentiation processes. The morphological and functional characterization of the model was carried out using optical microscopy, immunofluorescence, gene expression analysis by real-time PCR, and protein analysis by Western blot. The results confirmed the model’s capacity to reproduce adipogenesis and angiogenesis processes in an integrated manner, evidencing differences depending on the tissue origin. Subcutaneous adipose tissue showed a greater adipogenic capacity and three-dimensional organization, while epicardial adipose tissue exhibited a more inflammatory profile and a less cohesive structure. The angiogenic response was modest in both cases. Furthermore, structural and functional differences were observed between matrices, with Matrigel standing out as the most effective in forming stable spheroids. This model represents a useful preclinical tool to study the interaction between adipogenesis and angiogenesis processes in different tissues, as well as to explore new therapeutic approaches for cardiovascular diseases.
Direction
BARREIRO IGLESIAS, ANTON (Tutorships)
Eiras Penas, Sonia (Co-tutorships)
BARREIRO IGLESIAS, ANTON (Tutorships)
Eiras Penas, Sonia (Co-tutorships)
Court
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
Interpopulation variability in the level of photosynthetic inactivation due to heat stress in invasive species of the genus Carpobrotus
Authorship
M.D.M.C.S.
Bachelor of Biology
M.D.M.C.S.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The genus Carpobrotus includes various succulent species, most of which are native to South Africa. Due to their ornamental and stabilizing use, along with their deliberate propagation, some species of the genus have spread along the Atlantic-Mediterranean coasts throughout Europe and are now considered invasive. Their high phenotypic plasticity, hybridization capacity, clonal reproduction, and physiological integration favor their invasive success, displacing native species and compromising ecosystem biodiversity. Climate change and globalization have intensified these processes. The Carpobrotus spp. populations used, which include three genotypes (A, B, C, and hybrids), are distributed in native and introduced geographical regions with different climatic conditions. Therefore, to assess the interpopulation variability of the genus, photosynthetic inactivation thresholds were determined using fluorescence and reflectance measurements, in a temperature range between 35 and 60 degrees, in plants belonging to 11 Carpobrotus populations distributed in locations in South Africa, Italy, the US, New Zealand, Spain, and Chile. The results showed that Carpobrotus populations exhibit marked interpopulation variability in the level of photosynthetic inactivation, determined mainly by their genotype rather than their geographical origin. In addition, a thermal protective effect has been observed in all populations, which is activated when a threshold temperature is reached, generally between 50 and 55 degrees. Finally, at 60 degrees, acute thermal stress causes physiological collapse. These findings provide information that may be useful in delimiting the invasive potential of the genus in different climatic scenarios and contribute to the development of conservation strategies focused on the preservation of native species in vulnerable habitats. It is also essential to focus efforts on developing eradication and control plans targeting the most resilient populations of Carpobrotus spp. in their areas of introduction, especially in the face of increasing climate uncertainty.
The genus Carpobrotus includes various succulent species, most of which are native to South Africa. Due to their ornamental and stabilizing use, along with their deliberate propagation, some species of the genus have spread along the Atlantic-Mediterranean coasts throughout Europe and are now considered invasive. Their high phenotypic plasticity, hybridization capacity, clonal reproduction, and physiological integration favor their invasive success, displacing native species and compromising ecosystem biodiversity. Climate change and globalization have intensified these processes. The Carpobrotus spp. populations used, which include three genotypes (A, B, C, and hybrids), are distributed in native and introduced geographical regions with different climatic conditions. Therefore, to assess the interpopulation variability of the genus, photosynthetic inactivation thresholds were determined using fluorescence and reflectance measurements, in a temperature range between 35 and 60 degrees, in plants belonging to 11 Carpobrotus populations distributed in locations in South Africa, Italy, the US, New Zealand, Spain, and Chile. The results showed that Carpobrotus populations exhibit marked interpopulation variability in the level of photosynthetic inactivation, determined mainly by their genotype rather than their geographical origin. In addition, a thermal protective effect has been observed in all populations, which is activated when a threshold temperature is reached, generally between 50 and 55 degrees. Finally, at 60 degrees, acute thermal stress causes physiological collapse. These findings provide information that may be useful in delimiting the invasive potential of the genus in different climatic scenarios and contribute to the development of conservation strategies focused on the preservation of native species in vulnerable habitats. It is also essential to focus efforts on developing eradication and control plans targeting the most resilient populations of Carpobrotus spp. in their areas of introduction, especially in the face of increasing climate uncertainty.
Direction
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Tutorships)
Rodríguez Parra, Jonatan (Co-tutorships)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Tutorships)
Rodríguez Parra, Jonatan (Co-tutorships)
Court
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
Abiotic reactions using cellular machinery
Authorship
I.C.L.
Double bachelor degree in Chemistry and Biology (2ªed)
I.C.L.
Double bachelor degree in Chemistry and Biology (2ªed)
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Since the 2000s, one area of modern chemistry has experienced astonishing growth: bioorthogonal chemistry. This refers to any kind of chemical reaction that can be carried out within a living organism without affecting its metabolism. Bioorthogonality, in addition to avoiding secondary interactions with native functional groups, is characterized by high selectivity, rapid kinetics, and lack of toxicity. Consequently, the wide range of applications we find in biochemistry, biosynthesis, medicine and pharmacology is not surprising. Following the same philosophy, attempts are being made to develop reactions that are not only bioorthogonal but also based on photocatalytic processes. This would give rise to what we call Bioorthogonal Photocatalysis. In this Degree Final Project, new biocompatible synthetic reactions are investigated. Specifically, a photocatalytic reaction based on the a-arylation of enol acetates promoted by visible light and a photosensitizer. This reaction has been optimized, demonstrating its compatibility with aqueous media, an essential requirement for bioorthogonal chemistry. In order to transfer this transformation to cellular environments we have selected cyanobacteria since they have the necessary cellular machinery to catalyze the reaction with no external photocatalyst. This would allow the in-situ synthesis of drugs, bioactive compounds or fluorescent probes. Finally, efforts have been made to modify the substrate in order to obtain a fluorescent product, a key feature for applications in live-cell studies
Since the 2000s, one area of modern chemistry has experienced astonishing growth: bioorthogonal chemistry. This refers to any kind of chemical reaction that can be carried out within a living organism without affecting its metabolism. Bioorthogonality, in addition to avoiding secondary interactions with native functional groups, is characterized by high selectivity, rapid kinetics, and lack of toxicity. Consequently, the wide range of applications we find in biochemistry, biosynthesis, medicine and pharmacology is not surprising. Following the same philosophy, attempts are being made to develop reactions that are not only bioorthogonal but also based on photocatalytic processes. This would give rise to what we call Bioorthogonal Photocatalysis. In this Degree Final Project, new biocompatible synthetic reactions are investigated. Specifically, a photocatalytic reaction based on the a-arylation of enol acetates promoted by visible light and a photosensitizer. This reaction has been optimized, demonstrating its compatibility with aqueous media, an essential requirement for bioorthogonal chemistry. In order to transfer this transformation to cellular environments we have selected cyanobacteria since they have the necessary cellular machinery to catalyze the reaction with no external photocatalyst. This would allow the in-situ synthesis of drugs, bioactive compounds or fluorescent probes. Finally, efforts have been made to modify the substrate in order to obtain a fluorescent product, a key feature for applications in live-cell studies
Direction
Mascareñas Cid, Jose Luis (Tutorships)
TOMAS GAMASA, MARIA (Co-tutorships)
Mascareñas Cid, Jose Luis (Tutorships)
TOMAS GAMASA, MARIA (Co-tutorships)
Court
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
Multilevel selection theory: conflict between units selection
Authorship
I.C.L.
Double bachelor degree in Chemistry and Biology (2ªed)
I.C.L.
Double bachelor degree in Chemistry and Biology (2ªed)
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Following the acceptance of a theory of evolution mediated by natural selection and the recognition of the Modern Evolutionary Synthesis, the individual has been considered the level at which selection acts. Historically, almost every alternative has been questioned, turning this paradigm into one of the greatest pillars of Darwinism. This work reviews the most relevant literature on one of the most promising hypotheses in evolutionary biology: Multilevel Selection. This philosophy proposes that we abandon the simple model of natural selection intervening on the organism, since selection acts at different levels of biological organization, both below (cells, organelles, genes, etc.) and above (families, species, communities, etc.) the individual. The objective is to review the empirical evidence for selection proceeding at various levels and to study the biological conflict that would arise if selection at a higher level is opposed to selection at a lower level and vice versa. In addition, Multilevel Selection will be considered as a theoretical framework to explain one of the greatest enigmas of evolution: altruism. The methodology used includes the use of databases such as Web of Science and PubMed, and filters by language and date, prioritizing peer-reviewed studies. Our conclusions are positive, despite the fact that this hypothesis has been fraught with criticism since its inception and has been labeled a weak mechanism, ignoring much of the evidence. Over time, Multilevel Selection will be considered one of the most revolutionary ideas and one that has most changed our evolutionary thinking.
Following the acceptance of a theory of evolution mediated by natural selection and the recognition of the Modern Evolutionary Synthesis, the individual has been considered the level at which selection acts. Historically, almost every alternative has been questioned, turning this paradigm into one of the greatest pillars of Darwinism. This work reviews the most relevant literature on one of the most promising hypotheses in evolutionary biology: Multilevel Selection. This philosophy proposes that we abandon the simple model of natural selection intervening on the organism, since selection acts at different levels of biological organization, both below (cells, organelles, genes, etc.) and above (families, species, communities, etc.) the individual. The objective is to review the empirical evidence for selection proceeding at various levels and to study the biological conflict that would arise if selection at a higher level is opposed to selection at a lower level and vice versa. In addition, Multilevel Selection will be considered as a theoretical framework to explain one of the greatest enigmas of evolution: altruism. The methodology used includes the use of databases such as Web of Science and PubMed, and filters by language and date, prioritizing peer-reviewed studies. Our conclusions are positive, despite the fact that this hypothesis has been fraught with criticism since its inception and has been labeled a weak mechanism, ignoring much of the evidence. Over time, Multilevel Selection will be considered one of the most revolutionary ideas and one that has most changed our evolutionary thinking.
Direction
VILAS PETEIRO, ROMAN (Tutorships)
VILAS PETEIRO, ROMAN (Tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Study of the impact of interleukin-6 on circadian rhythms: evaluation of neuropeptides from the suprachiasmatic nucleus and circadian clock gene expression.
Authorship
A.D.L.
Bachelor of Biology
A.D.L.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
Introduction: Interleukin-6 (IL-6) is a pleiotropic cytokine with a central role in immune regulation and energy metabolism. Its levels exhibit daily fluctuations that influence essential components of the circadian system, suggesting an integrative role in synchronizing biological rhythms and metabolic homeostasis. Although some of its functions vary by sex, the specific role of IL-6 in integrating the circadian and metabolic systems has not been fully explored yet. Hypothesis: It is proposed that IL-6 participates in circadian and metabolic regulation differentially by sex, affecting key mechanisms of physiological synchronization. Objectives: The primary objective is to elucidate the role of IL-6 in connecting circadian and metabolic cycles. To this end, the influence of synchronizing neuropeptides, particularly vasoactive intestinal peptide (VIP), will also be evaluated as potential mediators of this coordination, paying special attention to sex-specific differences. Methodology: Circadian and metabolic phenotyping was performed in control and IL-6-deficient (IL6KO) mice of both sexes. Molecular analyses were performed in muscle and hypothalamus to elucidate a possible communication pathway between circadian and metabolic signals mediated by this cytokine. Furthermore, the study of VIP allowed us to establish its key role in the synchronization of the suprachiasmatic nucleus (SCN). Results: IL-6 deficiency affects circadian locomotor activity rhythms in a sex-dependent manner, altering synchronization to light in males and the endogenous period under dark conditions in both sexes. These effects were associated with a decrease in VIP in the SCN in IL6KO males, while levels remained stable in females. These findings, together with the results of molecular analyses in tissues, position IL-6 as a key mediator in circadian plasticity and its sex-dependent regulation, highlighting it as a potential therapeutic target for treating disorders associated with circadian misalignment.
Introduction: Interleukin-6 (IL-6) is a pleiotropic cytokine with a central role in immune regulation and energy metabolism. Its levels exhibit daily fluctuations that influence essential components of the circadian system, suggesting an integrative role in synchronizing biological rhythms and metabolic homeostasis. Although some of its functions vary by sex, the specific role of IL-6 in integrating the circadian and metabolic systems has not been fully explored yet. Hypothesis: It is proposed that IL-6 participates in circadian and metabolic regulation differentially by sex, affecting key mechanisms of physiological synchronization. Objectives: The primary objective is to elucidate the role of IL-6 in connecting circadian and metabolic cycles. To this end, the influence of synchronizing neuropeptides, particularly vasoactive intestinal peptide (VIP), will also be evaluated as potential mediators of this coordination, paying special attention to sex-specific differences. Methodology: Circadian and metabolic phenotyping was performed in control and IL-6-deficient (IL6KO) mice of both sexes. Molecular analyses were performed in muscle and hypothalamus to elucidate a possible communication pathway between circadian and metabolic signals mediated by this cytokine. Furthermore, the study of VIP allowed us to establish its key role in the synchronization of the suprachiasmatic nucleus (SCN). Results: IL-6 deficiency affects circadian locomotor activity rhythms in a sex-dependent manner, altering synchronization to light in males and the endogenous period under dark conditions in both sexes. These effects were associated with a decrease in VIP in the SCN in IL6KO males, while levels remained stable in females. These findings, together with the results of molecular analyses in tissues, position IL-6 as a key mediator in circadian plasticity and its sex-dependent regulation, highlighting it as a potential therapeutic target for treating disorders associated with circadian misalignment.
Direction
BARCA MAYO, OLGA (Tutorships)
GONZALEZ VILA, ANTIA (Co-tutorships)
BARCA MAYO, OLGA (Tutorships)
GONZALEZ VILA, ANTIA (Co-tutorships)
Court
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
New mechanisms involved in metabolic dysfunction-associated steatohepatitis: potential role of ANGPTL4
Authorship
P.D.O.
Bachelor of Biology
P.D.O.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the leading causes of chronic liver disease. Within the spectrum of this condition lies liver fibrosis, characterized by the activation of hepatic stellate cells (HSCs). The ANGPTL4 protein is an important regulator of hepatic lipid metabolism, but its role in HSCs is not clearly defined. This study analyzed the expression of ANGPTL4 in activated hepatic stellate cells, evaluated the effect of its silencing after stimulation with TGF-B1, and examined the impact of this silencing in a diet-induced murine model of steatohepatitis. The results showed that ANGPTL4 is overexpressed upon activation of hepatic stellate cells, and that its silencing reduces the expression of profibrotic genes, both in vitro and in vivo. These findings suggest that ANGPTL4 plays a pro-fibrotic role and could be a therapeutic target in MASLD.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is one of the leading causes of chronic liver disease. Within the spectrum of this condition lies liver fibrosis, characterized by the activation of hepatic stellate cells (HSCs). The ANGPTL4 protein is an important regulator of hepatic lipid metabolism, but its role in HSCs is not clearly defined. This study analyzed the expression of ANGPTL4 in activated hepatic stellate cells, evaluated the effect of its silencing after stimulation with TGF-B1, and examined the impact of this silencing in a diet-induced murine model of steatohepatitis. The results showed that ANGPTL4 is overexpressed upon activation of hepatic stellate cells, and that its silencing reduces the expression of profibrotic genes, both in vitro and in vivo. These findings suggest that ANGPTL4 plays a pro-fibrotic role and could be a therapeutic target in MASLD.
Direction
NOGUEIRAS POZO, RUBEN (Tutorships)
Parracho Martínez, Tamara (Co-tutorships)
NOGUEIRAS POZO, RUBEN (Tutorships)
Parracho Martínez, Tamara (Co-tutorships)
Court
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
Validation of biomarkers of response to tofacitinib treatment in patients with ulcerative colitis
Authorship
A.F.V.
Bachelor of Biology
A.F.V.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Tofacitinib, a Janus kinase (JAK) inhibitor, is the first small molecule administered orally that has been approved for the treatment of inflammatory bowel disease. The efficacy and safety of this compound make it an attractive therapeutic option for patients with moderate to severe forms of the disease. However, as with other treatments for inflammatory bowel disease, a significant percentage of patients do not respond to tofacitinib therapy. To date, there is no reliable biomarker available to predict the response to tofacitinib treatment, even though this would provide substantial medical and socioeconomic benefits. For this reason, the main objective of this project is to identify potential biomarkers of response to tofacitinib therapy and to optimize the experimental conditions for their subsequent validation. This proposal aims to lay the groundwork for the personalized treatment of patients with inflammatory bowel disease, thereby improving patient care. Peripheral blood mononuclear cell samples from patients with ulcerative colitis who responded to tofacitinib treatment were used. Through bioinformatic analysis of the samples, three potential biomarkers for tofacitinib treatment were identified: the genes KCNH8, RSAD2, and SGO2. In addition, the experimental conditions for the validation of these biomarkers in primary human cell cultures were established. It was determined that future validation will involve treatment with 10 ng/ml of interleukin 6 (IL-6) for 5 minutes and 50 nM of tofacitinib for 2 hours. All these steps, together with the subsequent validation of the biomarkers in future studies, will allow the evaluation of the potential of KCNH8, RSAD2, and SGO2 as markers of response to tofacitinib treatment in patients with ulcerative colitis, thus enabling individualized decision-making during the administration of this therapy in patients with ulcerative colitis.
Tofacitinib, a Janus kinase (JAK) inhibitor, is the first small molecule administered orally that has been approved for the treatment of inflammatory bowel disease. The efficacy and safety of this compound make it an attractive therapeutic option for patients with moderate to severe forms of the disease. However, as with other treatments for inflammatory bowel disease, a significant percentage of patients do not respond to tofacitinib therapy. To date, there is no reliable biomarker available to predict the response to tofacitinib treatment, even though this would provide substantial medical and socioeconomic benefits. For this reason, the main objective of this project is to identify potential biomarkers of response to tofacitinib therapy and to optimize the experimental conditions for their subsequent validation. This proposal aims to lay the groundwork for the personalized treatment of patients with inflammatory bowel disease, thereby improving patient care. Peripheral blood mononuclear cell samples from patients with ulcerative colitis who responded to tofacitinib treatment were used. Through bioinformatic analysis of the samples, three potential biomarkers for tofacitinib treatment were identified: the genes KCNH8, RSAD2, and SGO2. In addition, the experimental conditions for the validation of these biomarkers in primary human cell cultures were established. It was determined that future validation will involve treatment with 10 ng/ml of interleukin 6 (IL-6) for 5 minutes and 50 nM of tofacitinib for 2 hours. All these steps, together with the subsequent validation of the biomarkers in future studies, will allow the evaluation of the potential of KCNH8, RSAD2, and SGO2 as markers of response to tofacitinib treatment in patients with ulcerative colitis, thus enabling individualized decision-making during the administration of this therapy in patients with ulcerative colitis.
Direction
BALBOA MENDEZ, SABELA (Tutorships)
Conde Aranda, Javier (Co-tutorships)
BALBOA MENDEZ, SABELA (Tutorships)
Conde Aranda, Javier (Co-tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Metabolic Effect of Central Administration of a Pyk2 Inhibitor in Obese Rodents
Authorship
M.F.A.
Biotechnology Degree (2nd Ed. )
M.F.A.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
In recent decades, obesity has become a serious public health problem worldwide, making the identification of new targets that could lead to novel therapeutic treatments critically important. Previous studies have determined that Pyk2 is involved in various diseases in which symptoms are associated with disruptions in energy balance. However, its role in obesity has not been thoroughly investigated. Therefore, the aim of this study is to examine the effect of central administration of a Pyk2 inhibitor on energy balance in obese mice. Our results show a reduction in body weight in these mice, which is explained by a decrease in food intake following intracerebroventricular administration of a Pyk2 inhibitor. In white adipose tissue and the liver, lipid metabolism and B-oxidation pathways exhibit a strong compensatory mechanism in response to weight loss. However, at the molecular level, an increase in Badr3 receptor expression was detected in white adipose tissue, along with a decrease in PPARg expression in the liver. Nonetheless, chronic administration of the Pyk2 inhibitor does not induce an increase in thermogenesis markers in BAT. Altogether, our findings provide the first experimental evidence pointing to a key role of Pyk2 in the central nervous system in the regulation of energy balance.
In recent decades, obesity has become a serious public health problem worldwide, making the identification of new targets that could lead to novel therapeutic treatments critically important. Previous studies have determined that Pyk2 is involved in various diseases in which symptoms are associated with disruptions in energy balance. However, its role in obesity has not been thoroughly investigated. Therefore, the aim of this study is to examine the effect of central administration of a Pyk2 inhibitor on energy balance in obese mice. Our results show a reduction in body weight in these mice, which is explained by a decrease in food intake following intracerebroventricular administration of a Pyk2 inhibitor. In white adipose tissue and the liver, lipid metabolism and B-oxidation pathways exhibit a strong compensatory mechanism in response to weight loss. However, at the molecular level, an increase in Badr3 receptor expression was detected in white adipose tissue, along with a decrease in PPARg expression in the liver. Nonetheless, chronic administration of the Pyk2 inhibitor does not induce an increase in thermogenesis markers in BAT. Altogether, our findings provide the first experimental evidence pointing to a key role of Pyk2 in the central nervous system in the regulation of energy balance.
Direction
QUIÑONES TELLEZ, MARIA DEL MAR (Tutorships)
AL-MASSADI IGLESIAS, OMAR (Co-tutorships)
QUIÑONES TELLEZ, MARIA DEL MAR (Tutorships)
AL-MASSADI IGLESIAS, OMAR (Co-tutorships)
Court
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
Role of SIRT3 in AgRP Neurons in the Regulation of Glucose Metabolism.
Authorship
P.F.A.
Biotechnology Degree (2nd Ed. )
P.F.A.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 09:00
07.17.2025 09:00
Summary
Sirtuin 3 (SIRT3) is a well-known mitochondrial energy sensor that plays a key role in multiple biological processes, including energy and glucose metabolism. Although hypothalamic SIRT3 has been shown to regulate energy homeostasis, its specific role in glucose control and insulin sensitivity remains poorly understood. In this study, we investigated the effect of SIRT3 modulation in AgRP (agouti-related protein) neurons of the hypothalamus on glucose homeostasis and insulin sensitivity. To this end, we generated mouse models with loss of function of SIRT3 in AgRP neurons in order to characterize glucose metabolism and study its molecular mechanism of action. We observed that ablation of SIRT3 in AgRP neurons promoted glucose intolerance and reduced insulin sensitivity in both lean male mice and diet-induced obese mice. These effects are mediated by a worsening of AKT signaling in the liver, muscle, and white adipose tissue. Taken together, our findings establish a protective role of SIRT3 in AgRP neurons, mediated by AKT, in controlling obesity-induced glucose alterations.
Sirtuin 3 (SIRT3) is a well-known mitochondrial energy sensor that plays a key role in multiple biological processes, including energy and glucose metabolism. Although hypothalamic SIRT3 has been shown to regulate energy homeostasis, its specific role in glucose control and insulin sensitivity remains poorly understood. In this study, we investigated the effect of SIRT3 modulation in AgRP (agouti-related protein) neurons of the hypothalamus on glucose homeostasis and insulin sensitivity. To this end, we generated mouse models with loss of function of SIRT3 in AgRP neurons in order to characterize glucose metabolism and study its molecular mechanism of action. We observed that ablation of SIRT3 in AgRP neurons promoted glucose intolerance and reduced insulin sensitivity in both lean male mice and diet-induced obese mice. These effects are mediated by a worsening of AKT signaling in the liver, muscle, and white adipose tissue. Taken together, our findings establish a protective role of SIRT3 in AgRP neurons, mediated by AKT, in controlling obesity-induced glucose alterations.
Direction
QUIÑONES TELLEZ, MARIA DEL MAR (Tutorships)
AL-MASSADI IGLESIAS, OMAR (Co-tutorships)
QUIÑONES TELLEZ, MARIA DEL MAR (Tutorships)
AL-MASSADI IGLESIAS, OMAR (Co-tutorships)
Court
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Diatoms as indicators of emerging contaminants in freshwater ecosystems
Authorship
U.F.C.
Bachelor of Biology
U.F.C.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
Freshwater ecosystems are seriously threatened by anthropogenic impact, with emerging contaminants currently being one of the main concerns. Traditionally, water quality assessment has been based solely on physicochemical parameters, but in recent decades, the characterization of ecological status has been promoted as a more realistic perspective on the degree of ecosystem disturbance. This approach makes it necessary to use bioindicator organisms, among which diatoms have gained particular relevance due to their sensitivity to pollution, taxonomic diversity, and ubiquity. This study assesses the potential and current challenges of using diatoms as a tool for the detection and monitoring of emerging contaminants. To this end, a systematic bibliographic review was carried out using the Web of Science and Scopus databases. Several simple bibliometric analyses were conducted, along with an exploration of the existing knowledge in the fields of emerging contaminants and the use of diatoms as bioindicators. A growing interest was found in the use of diatoms, especially in lotic environments and for the study of pharmaceuticals as contaminants, particularly antibiotics. These findings highlight the increasing potential of diatoms as bioindicators of emerging contaminants, although their effective implementation still requires overcoming significant methodological challenges and developing new specific approaches for these pollutants.
Freshwater ecosystems are seriously threatened by anthropogenic impact, with emerging contaminants currently being one of the main concerns. Traditionally, water quality assessment has been based solely on physicochemical parameters, but in recent decades, the characterization of ecological status has been promoted as a more realistic perspective on the degree of ecosystem disturbance. This approach makes it necessary to use bioindicator organisms, among which diatoms have gained particular relevance due to their sensitivity to pollution, taxonomic diversity, and ubiquity. This study assesses the potential and current challenges of using diatoms as a tool for the detection and monitoring of emerging contaminants. To this end, a systematic bibliographic review was carried out using the Web of Science and Scopus databases. Several simple bibliometric analyses were conducted, along with an exploration of the existing knowledge in the fields of emerging contaminants and the use of diatoms as bioindicators. A growing interest was found in the use of diatoms, especially in lotic environments and for the study of pharmaceuticals as contaminants, particularly antibiotics. These findings highlight the increasing potential of diatoms as bioindicators of emerging contaminants, although their effective implementation still requires overcoming significant methodological challenges and developing new specific approaches for these pollutants.
Direction
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Tutorships)
LEIRA CAMPOS, ANTON MANOEL (Co-tutorships)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Tutorships)
LEIRA CAMPOS, ANTON MANOEL (Co-tutorships)
Court
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
Lighting colour temperature effect on architectural heritage biocolonization: an experimental study with a microalgae model strain
Authorship
C.F.G.
Bachelor of Biology
C.F.G.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Architectural heritage undergoes biological colonization processes, which in temperate and humid climates correspond mainly to green algae and cyanobacteria (phototrophic organisms) that form subaerial biofilms (SABs) on surfaces. The use of ornamental lighting with Light Emitting Diodes (LEDs) is a common practice to improve the visibility and aesthetics of the heritage, and it has been proven that this lighting affects the colonization of phototrophs that depend on light for photosynthetic processes. Nocturnal ornamental lighting can have different color temperatures, from lower and warmer (2000K-3000K) with orange hues, to higher and cooler (6000K-7000K) with bluish hues. To evaluate their impact on the development of phototrophs on surfaces of heritage buildings (leading to their biodeterioration), studied the effect of three color temperatures, 6000K (cool white), 4000K (neutral white) and 2700K (warm white) on the growth of biofilms of the model microalgae Jaagichlorella sp., common in Central European buildings. The biofilms were developed to maturity (38 days) on polycarbonate membranes under these three illuminations and without ornamental light (control). Finally, the photosynthetic pigment content was determined and a pulse amplitude modulation (PAM) fluorometer was used to observe the photosynthetic performance of each biofilm. The results indicated increased pigment content in biofilms grown under the 6000K and 4000K lights, and higher wet biomass and photosynthetic efficiency under the 2700K light. It is inferred that, lights with high color temperatures (bluish) cause a higher physiological stress to Jaagichlorella sp. cells reducing their growth in biofilm form, while lights with lower color temperatures (orange) promote growth, which transferred to heritage conservation against biodeterioration that could cause Jaagichlorella sp. makes avoiding a warm lighting against a cold one.
Architectural heritage undergoes biological colonization processes, which in temperate and humid climates correspond mainly to green algae and cyanobacteria (phototrophic organisms) that form subaerial biofilms (SABs) on surfaces. The use of ornamental lighting with Light Emitting Diodes (LEDs) is a common practice to improve the visibility and aesthetics of the heritage, and it has been proven that this lighting affects the colonization of phototrophs that depend on light for photosynthetic processes. Nocturnal ornamental lighting can have different color temperatures, from lower and warmer (2000K-3000K) with orange hues, to higher and cooler (6000K-7000K) with bluish hues. To evaluate their impact on the development of phototrophs on surfaces of heritage buildings (leading to their biodeterioration), studied the effect of three color temperatures, 6000K (cool white), 4000K (neutral white) and 2700K (warm white) on the growth of biofilms of the model microalgae Jaagichlorella sp., common in Central European buildings. The biofilms were developed to maturity (38 days) on polycarbonate membranes under these three illuminations and without ornamental light (control). Finally, the photosynthetic pigment content was determined and a pulse amplitude modulation (PAM) fluorometer was used to observe the photosynthetic performance of each biofilm. The results indicated increased pigment content in biofilms grown under the 6000K and 4000K lights, and higher wet biomass and photosynthetic efficiency under the 2700K light. It is inferred that, lights with high color temperatures (bluish) cause a higher physiological stress to Jaagichlorella sp. cells reducing their growth in biofilm form, while lights with lower color temperatures (orange) promote growth, which transferred to heritage conservation against biodeterioration that could cause Jaagichlorella sp. makes avoiding a warm lighting against a cold one.
Direction
SANMARTIN SANCHEZ, PATRICIA (Tutorships)
MENDEZ VILLAR, ANXO (Co-tutorships)
SANMARTIN SANCHEZ, PATRICIA (Tutorships)
MENDEZ VILLAR, ANXO (Co-tutorships)
Court
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
Effect of partial chemical reprogramming on progeria
Authorship
B.F.M.
Biotechnology Degree (2nd Ed. )
B.F.M.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Hutchinson Gilford Progeria Syndrome is caused by the accumulation of farnesylated lamin A at the nuclear envelope, which leads to a prematurely aged phenotype in cells. This phenotype is identified by cell cycle arrest, abnormal nuclear morphology and senescence identity. Several strategies have been proposed to reverse cellular aging, focusing in partial reprogramming, among which the use of small chemical compounds such as tranylcypromine and RepSox stand out. These molecules act on an epigenetic level as well as on the transforming growth factor beta signalling pathway, thereby modifying gene expression and improving the aged phenotype. This study aims to evaluate the effectiveness of this chemical compounds in human dermal fibroblast that exhibit lamin A accumulation on the nuclear envelope. To do this, senescence and proliferative assays were performed to determine successful disease induction and subsequently to assess the effects of the proposes treatment. In addition, a genetic expression analysis is carried out to determine the effect of the treatment on epigenetic markers. Disease induction was successful, as demonstrated by clonogenic assays and microscopic observations. Moreover, treatment with tranylcypromine and RepSox proved beneficial, as evidenced by improvement in senescence markers, both genetic (CDKN1A, GDF-15 and SERPINE-1) and enzymatic (galactosidase activity) after a week of treatment.
Hutchinson Gilford Progeria Syndrome is caused by the accumulation of farnesylated lamin A at the nuclear envelope, which leads to a prematurely aged phenotype in cells. This phenotype is identified by cell cycle arrest, abnormal nuclear morphology and senescence identity. Several strategies have been proposed to reverse cellular aging, focusing in partial reprogramming, among which the use of small chemical compounds such as tranylcypromine and RepSox stand out. These molecules act on an epigenetic level as well as on the transforming growth factor beta signalling pathway, thereby modifying gene expression and improving the aged phenotype. This study aims to evaluate the effectiveness of this chemical compounds in human dermal fibroblast that exhibit lamin A accumulation on the nuclear envelope. To do this, senescence and proliferative assays were performed to determine successful disease induction and subsequently to assess the effects of the proposes treatment. In addition, a genetic expression analysis is carried out to determine the effect of the treatment on epigenetic markers. Disease induction was successful, as demonstrated by clonogenic assays and microscopic observations. Moreover, treatment with tranylcypromine and RepSox proved beneficial, as evidenced by improvement in senescence markers, both genetic (CDKN1A, GDF-15 and SERPINE-1) and enzymatic (galactosidase activity) after a week of treatment.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
Court
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
Applications of seaweed-derived phycocolloids in industry
Authorship
Y.F.P.
Bachelor of Biology
Y.F.P.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
Phycocolloids are polysaccharides located in the cell wall of red and brown macroalgae, with the ability to alter the rheological properties of the aqueous solutions in which they are found. This bibliographic review focuses on the study of the three most important phycocolloids in the industry (carrageenans, agar and alginate) and their role across various industrial sectors. In this context, characteristics such as their structure and properties are analyzed, as these are directly related to their activities and applications. Furthermore, the potential of these compounds is explored, which has enabled their incorporation into a variety of industrial sectors, including the food, pharmaceutical, and cosmetic industries, among others. In addition, the current state of the phycocolloid industry is analyzed, and its future prospects are discussed. The remarkable potential of phycocolloids in the industry is evident. This work highlights the growing interest in their research and the increasing demand for products based on these compounds. However, the current pace of industrial development is not aligned with this potential, creating a challenging scenario for the future of phycocolloids in the sector.
Phycocolloids are polysaccharides located in the cell wall of red and brown macroalgae, with the ability to alter the rheological properties of the aqueous solutions in which they are found. This bibliographic review focuses on the study of the three most important phycocolloids in the industry (carrageenans, agar and alginate) and their role across various industrial sectors. In this context, characteristics such as their structure and properties are analyzed, as these are directly related to their activities and applications. Furthermore, the potential of these compounds is explored, which has enabled their incorporation into a variety of industrial sectors, including the food, pharmaceutical, and cosmetic industries, among others. In addition, the current state of the phycocolloid industry is analyzed, and its future prospects are discussed. The remarkable potential of phycocolloids in the industry is evident. This work highlights the growing interest in their research and the increasing demand for products based on these compounds. However, the current pace of industrial development is not aligned with this potential, creating a challenging scenario for the future of phycocolloids in the sector.
Direction
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Tutorships)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Tutorships)
Court
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
Analysis of the neurological phenotype in mice with Nae1 deletion in astrocytes: effects on neuronal and astrocytic function.
Authorship
B.F.R.
Bachelor of Biology
B.F.R.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
Astrocytes are essential cells for maintaining brain homeostasis, neurotransmission, and the integrity of the blood-brain barrier. They also play a key role in the neuroinflammatory response, adopting reactive phenotypes to pathological stimuli. This paper studied a mouse model with an astrocytic mutation that induces neuroinflammation. Microglia, as the main immune cell of the central nervous system, participate in this response, amplifying the inflammatory effect. Neddylation is a post-translational modification that mediates the binding of the NEDD8 protein to target proteins. This mechanism controls processes such as stress response and protein degradation. The enzyme NAE1 initiates this cascade by activating NEDD8, which can modify the stability of target proteins and direct them to the proteasome. A conditional and inducible mouse model was generated with deletion of the Nae1 gene in astrocytes of the GLAST subpopulation. After recombination induction with tamoxifen, the mice had a lifespan of approximately 10 weeks and developed phenotypic alterations. Previous proteomic studies in the hypothalamus revealed elevated levels of proinflammatory cytokines. Therefore, the objective of this study was to determine the hypothalamic nuclei where neuroinflammation is located and to evaluate the integrity of the blood-brain barrier. Immunofluorescence was performed with antibodies against GFAP (a marker of astrogliosis), IBA1 (a marker of activated microglia), and CD45+ (a leukocyte marker). The results showed an elevation of all these markers in the mutant mice, indicating neuroinflammation and infiltration of leukocytes from the peripheral immune system, suggesting barrier alterations. The cause of this inflammation is undetermined, highlighting the need to understand the effect of Nae1 deletion on astrocytes. Other models with Nae1 deletions in adult neurons showed no apparent phenotypes. This study highlights the importance of both astrocytes and the neddylation process.
Astrocytes are essential cells for maintaining brain homeostasis, neurotransmission, and the integrity of the blood-brain barrier. They also play a key role in the neuroinflammatory response, adopting reactive phenotypes to pathological stimuli. This paper studied a mouse model with an astrocytic mutation that induces neuroinflammation. Microglia, as the main immune cell of the central nervous system, participate in this response, amplifying the inflammatory effect. Neddylation is a post-translational modification that mediates the binding of the NEDD8 protein to target proteins. This mechanism controls processes such as stress response and protein degradation. The enzyme NAE1 initiates this cascade by activating NEDD8, which can modify the stability of target proteins and direct them to the proteasome. A conditional and inducible mouse model was generated with deletion of the Nae1 gene in astrocytes of the GLAST subpopulation. After recombination induction with tamoxifen, the mice had a lifespan of approximately 10 weeks and developed phenotypic alterations. Previous proteomic studies in the hypothalamus revealed elevated levels of proinflammatory cytokines. Therefore, the objective of this study was to determine the hypothalamic nuclei where neuroinflammation is located and to evaluate the integrity of the blood-brain barrier. Immunofluorescence was performed with antibodies against GFAP (a marker of astrogliosis), IBA1 (a marker of activated microglia), and CD45+ (a leukocyte marker). The results showed an elevation of all these markers in the mutant mice, indicating neuroinflammation and infiltration of leukocytes from the peripheral immune system, suggesting barrier alterations. The cause of this inflammation is undetermined, highlighting the need to understand the effect of Nae1 deletion on astrocytes. Other models with Nae1 deletions in adult neurons showed no apparent phenotypes. This study highlights the importance of both astrocytes and the neddylation process.
Direction
BARCA MAYO, OLGA (Tutorships)
BARCA MAYO, OLGA (Tutorships)
Court
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
Effect of intraindividual variability on the photochemical performance of Fucus vesiculosus L. under desiccation.
Authorship
I.F.A.
Bachelor of Biology
I.F.A.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Macroalgae such as Fucus vesiculosus play a fundamental role in intertidal ecosystems, acting as ecosystem engineers under extreme environmetal conditions. Periodic desiccation associated with tidal cycles represents a significant physiological challenge, potentially compromising photosynthetic activity and cell viability. This study proposes that Fucus vesiculosus exhibits intraindividual functional heterogeneity: apical tissues (more recent) would respond better to stress than basal tissues (more mature). To test this, chlorophyll fluorescence parameters were analyzed in three consecutive dichotomies of the thallus. Statistical analysis revealed significant differences between the dichotomies, with higher photochemical efficiency and lower non-regulated energy dissipation in the apical tissues, indicating greater physiological resilience. In contrast, basal tissues showed more limited photosynthetic performance. These results support the existence of a functional gradient associated with tissue age and position, and suggest that Fucus vesiculosus may employ an adaptative strategy based on the preferential protection of its youngest segments. This study contributes to the understanding of macroalgal physiology and provides new evidence regarding their adaptative strategies in response to increasing environmental stress driven by climate change in intertidal ecosystems.
Macroalgae such as Fucus vesiculosus play a fundamental role in intertidal ecosystems, acting as ecosystem engineers under extreme environmetal conditions. Periodic desiccation associated with tidal cycles represents a significant physiological challenge, potentially compromising photosynthetic activity and cell viability. This study proposes that Fucus vesiculosus exhibits intraindividual functional heterogeneity: apical tissues (more recent) would respond better to stress than basal tissues (more mature). To test this, chlorophyll fluorescence parameters were analyzed in three consecutive dichotomies of the thallus. Statistical analysis revealed significant differences between the dichotomies, with higher photochemical efficiency and lower non-regulated energy dissipation in the apical tissues, indicating greater physiological resilience. In contrast, basal tissues showed more limited photosynthetic performance. These results support the existence of a functional gradient associated with tissue age and position, and suggest that Fucus vesiculosus may employ an adaptative strategy based on the preferential protection of its youngest segments. This study contributes to the understanding of macroalgal physiology and provides new evidence regarding their adaptative strategies in response to increasing environmental stress driven by climate change in intertidal ecosystems.
Direction
ABOAL VIÑAS, JESUS RAMON (Tutorships)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Co-tutorships)
ABOAL VIÑAS, JESUS RAMON (Tutorships)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Co-tutorships)
Court
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
Quantification of seipin isoforms in primary fibroblasts from patients with congenital generalized lipodystrophy type 2
Authorship
R.F.T.
Bachelor of Biology
R.F.T.
Bachelor of Biology
Defense date
07.16.2025 09:30
07.16.2025 09:30
Summary
Lipodystrophies are a heterogeneous group of rare diseases characterized by the loss of adipose tissue and other associated metabolic alterations. Among them, one of the most severe forms is congenital generalized lipodystrophy type 2, caused by variants in the BSCL2 gene, which encodes the protein seipin. A particularly severe subtype is Celia’s encephalopathy, a neurodegenerative disease caused by the presence of an aberrant isoform of seipin, known as Celia seipin. In order to optimize the detection protocol for this aberrant form, as well as for the wild-type seipin, this study was designed-based on the initial hypothesis that, due to their characteristics, the antibodies used in the research should be effective in identifying this protein. For the development of this study, primary fibroblast cultures were used, obtained from skin biopsies of patients with this condition and from a healthy individual (control). From these cells, protein extractions and quantifications were performed using the Bradford method, followed by analysis through Western blotting on Tris-Gly and Bis-Tris gels. Finally, quantification of the bands corresponding to seipin was carried out using Image Lab software. In this way, the aim was to evaluate the detection capability of different antibodies, with results showing that most of them were not effective for detecting seipin. However, the anti-seipin antibody (Assay Genie) proved to be useful for detecting wild-type seipin. It is expected that all of this will contribute to the development and optimization of a protocol for detecting seipin in samples from patients with Celia’s encephalopathy, thus helping to advance research into this condition.
Lipodystrophies are a heterogeneous group of rare diseases characterized by the loss of adipose tissue and other associated metabolic alterations. Among them, one of the most severe forms is congenital generalized lipodystrophy type 2, caused by variants in the BSCL2 gene, which encodes the protein seipin. A particularly severe subtype is Celia’s encephalopathy, a neurodegenerative disease caused by the presence of an aberrant isoform of seipin, known as Celia seipin. In order to optimize the detection protocol for this aberrant form, as well as for the wild-type seipin, this study was designed-based on the initial hypothesis that, due to their characteristics, the antibodies used in the research should be effective in identifying this protein. For the development of this study, primary fibroblast cultures were used, obtained from skin biopsies of patients with this condition and from a healthy individual (control). From these cells, protein extractions and quantifications were performed using the Bradford method, followed by analysis through Western blotting on Tris-Gly and Bis-Tris gels. Finally, quantification of the bands corresponding to seipin was carried out using Image Lab software. In this way, the aim was to evaluate the detection capability of different antibodies, with results showing that most of them were not effective for detecting seipin. However, the anti-seipin antibody (Assay Genie) proved to be useful for detecting wild-type seipin. It is expected that all of this will contribute to the development and optimization of a protocol for detecting seipin in samples from patients with Celia’s encephalopathy, thus helping to advance research into this condition.
Direction
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ARAUJO VILAR, DAVID (Co-tutorships)
SANCHEZ IGLESIAS, SOFIA (Co-tutorships)
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ARAUJO VILAR, DAVID (Co-tutorships)
SANCHEZ IGLESIAS, SOFIA (Co-tutorships)
Court
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
Antibacterial activity of synthetic hydrogels derived from (-)-shikimic acid with encapsulated antibiotics
Authorship
S.F.L.
Double bachelor degree in Chemistry and Biology
S.F.L.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
Interest in hydrogels has increased in recent years due to their biocompatible properties, positioning them as materials of great projection in biotechnological industries such as pharmaceuticals and tissue engineering. Due to the inappropriate and abusive use of antimicrobial drugs, bacteria have developed multiple mechanisms of resistance to the action of antibiotics. In this Final Degree Project is proposed the use of a molecular hydrogel derived from (-)-shikimic acid as a transport and localized drug delivery system. Two main objectives were formulated: to evaluate the intrinsic antibacterial action of the gel, and to evaluate the capacity of encapsulation and release of antibiotics, by means of plaque diffusion studies with four bacterial species, three of them Gram-negative (Escherichia coli, Salmonella enterica subsp. enterica and Acinetobacter baumannii) and one Gram-positive (Staphylococcus epidermidis). In addition, the alteration of different parameters led to the formulation of two specific objectives: determination of the effect of incubation conditions (time and temperature), and the effect of the concentration of antibiotic used. In the latter case, the sensitivity of the bacteria was evaluated against two antibiotics from different families: beta-lactams and quinolones. The results ruled out the inherent antibacterial activity of the hydrogel but demonstrated its efficacy as an antibiotic delivery medium. Complete release of the antibiotic trapped in the hydrogel was observed, and higher concentrations resulted in greater diffusion on the plate. This verified its efficacy in inhibiting bacterial growth, with S. enterica being the bacterial species that showed the greatest sensitivity to both antibiotics. The results obtained suggest that newly synthesized hydrogels could be a promising route of antibiotic delivery in the face of the incessant emergence of bacterial resistance.
Interest in hydrogels has increased in recent years due to their biocompatible properties, positioning them as materials of great projection in biotechnological industries such as pharmaceuticals and tissue engineering. Due to the inappropriate and abusive use of antimicrobial drugs, bacteria have developed multiple mechanisms of resistance to the action of antibiotics. In this Final Degree Project is proposed the use of a molecular hydrogel derived from (-)-shikimic acid as a transport and localized drug delivery system. Two main objectives were formulated: to evaluate the intrinsic antibacterial action of the gel, and to evaluate the capacity of encapsulation and release of antibiotics, by means of plaque diffusion studies with four bacterial species, three of them Gram-negative (Escherichia coli, Salmonella enterica subsp. enterica and Acinetobacter baumannii) and one Gram-positive (Staphylococcus epidermidis). In addition, the alteration of different parameters led to the formulation of two specific objectives: determination of the effect of incubation conditions (time and temperature), and the effect of the concentration of antibiotic used. In the latter case, the sensitivity of the bacteria was evaluated against two antibiotics from different families: beta-lactams and quinolones. The results ruled out the inherent antibacterial activity of the hydrogel but demonstrated its efficacy as an antibiotic delivery medium. Complete release of the antibiotic trapped in the hydrogel was observed, and higher concentrations resulted in greater diffusion on the plate. This verified its efficacy in inhibiting bacterial growth, with S. enterica being the bacterial species that showed the greatest sensitivity to both antibiotics. The results obtained suggest that newly synthesized hydrogels could be a promising route of antibiotic delivery in the face of the incessant emergence of bacterial resistance.
Direction
LEMOS RAMOS, MANUEL LUIS (Tutorships)
AFONSO LAGES, MARTA CAROLINA (Co-tutorships)
LEMOS RAMOS, MANUEL LUIS (Tutorships)
AFONSO LAGES, MARTA CAROLINA (Co-tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Bolaamphiphilic alkylbisamide hydrogels derived from (-)-shikimic acid with odd-length hydrocarbonated spacer: New nanostructured materials for drug delivery
Authorship
S.F.L.
Double bachelor degree in Chemistry and Biology
S.F.L.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Low molecular weight gels stand out for their versatility and structural variety, as they are soft and flexible materials capable of trapping liquids within their network, and boasting excellent properties in terms of softness, biodegradability, and biocompatibility. These characteristics generate great interest in fields such as biomedicine, cosmetics, and the food industry, increasingly expanding its outreach. This Final Degree Project focuses on the study of these materials as nanostructured systems for the transport and controlled delivery of drugs. The objectives of the project include the synthesis of a molecule derived from commercially available (-)-shikimic acid and the evaluation of its potential gelation capacity in various solvents. Furthermore, the study of its hydrogels that show optimal properties for their application as “drug delivery systems”, specifically for the entrapment and release of broad-spectrum antibiotics, is further explored.
Low molecular weight gels stand out for their versatility and structural variety, as they are soft and flexible materials capable of trapping liquids within their network, and boasting excellent properties in terms of softness, biodegradability, and biocompatibility. These characteristics generate great interest in fields such as biomedicine, cosmetics, and the food industry, increasingly expanding its outreach. This Final Degree Project focuses on the study of these materials as nanostructured systems for the transport and controlled delivery of drugs. The objectives of the project include the synthesis of a molecule derived from commercially available (-)-shikimic acid and the evaluation of its potential gelation capacity in various solvents. Furthermore, the study of its hydrogels that show optimal properties for their application as “drug delivery systems”, specifically for the entrapment and release of broad-spectrum antibiotics, is further explored.
Direction
Estévez Cabanas, Juan Carlos (Tutorships)
QUIÑOA CABANA, EMILIO (Co-tutorships)
Estévez Cabanas, Juan Carlos (Tutorships)
QUIÑOA CABANA, EMILIO (Co-tutorships)
Court
SAA RODRIGUEZ, CARLOS EUGENIO (Chairman)
SOUSA PEDRARES, ANTONIO (Secretary)
GARCIA DEIBE, ANA MARIA (Member)
SAA RODRIGUEZ, CARLOS EUGENIO (Chairman)
SOUSA PEDRARES, ANTONIO (Secretary)
GARCIA DEIBE, ANA MARIA (Member)
Gene drives for pest control: a recap
Authorship
D.F.F.
Biotechnology Degree (2nd Ed. )
D.F.F.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
Pest control remains one of the main challenges for public health, agriculture, and biodi-versity conservation. In recent decades, the development of genetic technologies such as gene drives has opened new possibilities for the specific and long-term control of tar-get species. This work aims to provide a bibliographic review and critical analysis of pest control methods based on gene drive technology, comparing them to conventional strat-egies and evaluating their environmental and social implications. The study examines the main molecular mechanisms employed, the types of gene drives developed to date, their technical limitations, and the ecological risks associated with their release. It also compares their advantages and drawbacks with traditional methods such as pesticides and identifies key ethical, regulatory, and scientific challenges that affect their future implementation. The results of this review suggest that although gene drives offer significant potential as a control tool, their use entails unresolved technical challenges and considerable ecolog-ical uncertainty. It is concluded that their application should proceed under robust inter-national governance frameworks, guided by scientific evidence, the precautionary princi-ple, and active public engagement.
Pest control remains one of the main challenges for public health, agriculture, and biodi-versity conservation. In recent decades, the development of genetic technologies such as gene drives has opened new possibilities for the specific and long-term control of tar-get species. This work aims to provide a bibliographic review and critical analysis of pest control methods based on gene drive technology, comparing them to conventional strat-egies and evaluating their environmental and social implications. The study examines the main molecular mechanisms employed, the types of gene drives developed to date, their technical limitations, and the ecological risks associated with their release. It also compares their advantages and drawbacks with traditional methods such as pesticides and identifies key ethical, regulatory, and scientific challenges that affect their future implementation. The results of this review suggest that although gene drives offer significant potential as a control tool, their use entails unresolved technical challenges and considerable ecolog-ical uncertainty. It is concluded that their application should proceed under robust inter-national governance frameworks, guided by scientific evidence, the precautionary princi-ple, and active public engagement.
Direction
GARCIA SUAREZ, CARLOS (Tutorships)
GARCIA SUAREZ, CARLOS (Tutorships)
Court
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
Sirtuins, aging and caloric restriction
Authorship
C.F.B.
Bachelor of Biology
C.F.B.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Sirtuins are a family of deacetylase enzymes conserved from prokaryotes to eukaryotes. They were first described in Saccharomyces cerevisiae, and were attributed with the ability to extend the lifespan of this organism when overexpressed through interventions such as caloric restriction and genetic modification. In the years that followed, numerous experiments were conducted to determine whether the lifespan-extending effects of sirtuins in yeast could be translated to more complex organisms. The initial results seemed promising, and studies in humans even began. Nevertheless, more recent studies have cast doubt on the previously held views about the role of sirtuins in aging, with some findings being completely contradictory. Unfortunately, the initial excitement they generated led to a wave of review articles that now cloud the literature and introduce bias among researchers. There is still a long way to go. Sirtuins remain a topic of interest today as potential targets in molecular medicine, since, despite the uncertainty surrounding their impact on lifespan, they have proven useful in protecting organisms against various age-related conditions.
Sirtuins are a family of deacetylase enzymes conserved from prokaryotes to eukaryotes. They were first described in Saccharomyces cerevisiae, and were attributed with the ability to extend the lifespan of this organism when overexpressed through interventions such as caloric restriction and genetic modification. In the years that followed, numerous experiments were conducted to determine whether the lifespan-extending effects of sirtuins in yeast could be translated to more complex organisms. The initial results seemed promising, and studies in humans even began. Nevertheless, more recent studies have cast doubt on the previously held views about the role of sirtuins in aging, with some findings being completely contradictory. Unfortunately, the initial excitement they generated led to a wave of review articles that now cloud the literature and introduce bias among researchers. There is still a long way to go. Sirtuins remain a topic of interest today as potential targets in molecular medicine, since, despite the uncertainty surrounding their impact on lifespan, they have proven useful in protecting organisms against various age-related conditions.
Direction
BARJA FRANCISCO, PRIMITIVO (Tutorships)
BARJA FRANCISCO, PRIMITIVO (Tutorships)
Court
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
Design of new molecular systems for the stabilization of nucleic acids in deep eutectic solvents
Authorship
M.G.G.
Double bachelor degree in Chemistry and Biology
M.G.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Nucleic acids are currently used in a wide range of treatments due to their diverse types and functions, and they have the potential to become highly important drugs in the coming decades. One of their main drawbacks is their low stability in aqueous media, being susceptible to chemical hydrolysis, aggregation, and enzymatic degradation. A possible solution to this problem is the use of deep eutectic solvents, a type of non-aqueous solvent recently discovered, composed of a hydrogen bond donor and acceptor held together by intermolecular forces. They contain no water in their formulation, are thermally stable, inexpensive, non-toxic, and capable of dissolving biomolecules, making them potential candidates for stabilizing nucleic acids. In this research, their low toxicity was assessed through cell viability assays using MTT on model cell lines. The results obtained indicate low toxicity at relatively high concentrations, highlighting their biocompatibility and potential application in biological chemistry.
Nucleic acids are currently used in a wide range of treatments due to their diverse types and functions, and they have the potential to become highly important drugs in the coming decades. One of their main drawbacks is their low stability in aqueous media, being susceptible to chemical hydrolysis, aggregation, and enzymatic degradation. A possible solution to this problem is the use of deep eutectic solvents, a type of non-aqueous solvent recently discovered, composed of a hydrogen bond donor and acceptor held together by intermolecular forces. They contain no water in their formulation, are thermally stable, inexpensive, non-toxic, and capable of dissolving biomolecules, making them potential candidates for stabilizing nucleic acids. In this research, their low toxicity was assessed through cell viability assays using MTT on model cell lines. The results obtained indicate low toxicity at relatively high concentrations, highlighting their biocompatibility and potential application in biological chemistry.
Direction
MONTENEGRO GARCIA, JAVIER (Tutorships)
SANCHEZ FERNANDEZ, ADRIAN (Co-tutorships)
MONTENEGRO GARCIA, JAVIER (Tutorships)
SANCHEZ FERNANDEZ, ADRIAN (Co-tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Design of new molecular systems for the stabilization and delivery of nucleic acids in deep eutectic solvents
Authorship
M.G.G.
Double bachelor degree in Chemistry and Biology
M.G.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Deep eutectic solvents (DESs) are liquids formed by mixing a hydrogen bond donor and a hydrogen bond acceptor, which promote a depression in the melting point compared to their individual components. DESs are formed through a variety of intermolecular forces (hydrogen bonds, electrostatic interactions, van der Waals forces), where different combinations of precursors allow for the generation of DESs with varied properties. Furthermore, their stability, low toxicity, and lack of water make them promising candidates for stabilizing complex molecules. In this study, experiments were conducted on the solubilization and incorporation of nucleic acids into DESs. Initially, DESs with varied compositions were formulated, and their water content was assessed using the Karl Fischer method, along with infrared spectroscopy and proton nuclear magnetic resonance measurements to verify their structure. Transfer ribonucleic acid (tRNA) was incorporated into these DESs. The conformation of the tRNA was examined using ultraviolet spectroscopy, and degradation tests were carried out to assess the stabilizing properties of the DESs. The results show the proper formation of the DESs, their purity, and a low water content. Preliminary results on the solubilization and degradation of tRNA demonstrate the ability of DESs to contain and stabilize the biomolecule. This study highlights the potential of DESs for stabilizing labile biomolecules in non-aqueous media.
Deep eutectic solvents (DESs) are liquids formed by mixing a hydrogen bond donor and a hydrogen bond acceptor, which promote a depression in the melting point compared to their individual components. DESs are formed through a variety of intermolecular forces (hydrogen bonds, electrostatic interactions, van der Waals forces), where different combinations of precursors allow for the generation of DESs with varied properties. Furthermore, their stability, low toxicity, and lack of water make them promising candidates for stabilizing complex molecules. In this study, experiments were conducted on the solubilization and incorporation of nucleic acids into DESs. Initially, DESs with varied compositions were formulated, and their water content was assessed using the Karl Fischer method, along with infrared spectroscopy and proton nuclear magnetic resonance measurements to verify their structure. Transfer ribonucleic acid (tRNA) was incorporated into these DESs. The conformation of the tRNA was examined using ultraviolet spectroscopy, and degradation tests were carried out to assess the stabilizing properties of the DESs. The results show the proper formation of the DESs, their purity, and a low water content. Preliminary results on the solubilization and degradation of tRNA demonstrate the ability of DESs to contain and stabilize the biomolecule. This study highlights the potential of DESs for stabilizing labile biomolecules in non-aqueous media.
Direction
MONTENEGRO GARCIA, JAVIER (Tutorships)
SANCHEZ FERNANDEZ, ADRIAN (Co-tutorships)
MONTENEGRO GARCIA, JAVIER (Tutorships)
SANCHEZ FERNANDEZ, ADRIAN (Co-tutorships)
Court
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
Origin and evolution of bipedalism in the human lineage
Authorship
J.G.T.
Bachelor of Biology
J.G.T.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The origin of bipedalism and its consolidation as the primary locomotor system in humans has been subject of study in recent decades thanks to the increasing discovery and analysis of the fossil record corresponding to the different taxa that made up the human lineage. Anatomical comparison with the great apes that coexist with us today allow us to identify the characteristics that differentiate orthograde bipedalism from quadrupedal locomotion and pronograde posture. Thus, the widening and shortening of the pelvis, the development of lumbar lordosis, and the modification of the foot to a purely support structure are anatomical changes resulting from a evolutionary process subject to a combination of multiple selective pressures and localized in open environments with a significant presence of trees. The fossil record shows the course of this evolution from the appearance of the first signs of facultative bipedalism in genera such as Orrorin and Ardipithecus. The genus Australopithecus presents a mosaic of arboreal and bipedal traits, with habitual bipedalism, and represents a turning point in the mode of locomotion of the human lineage. Finally, bipedal locomotion is now considered obligatory in the genus Homo, as the characteristics suggesting arboreal behavior become minimal. However, the morphology of this genus is also marked by strong sexual dimorphism resulting from the evolutionary force of obstetric difficulties caused by the expansion of the braincase and the narrowing of the birth canal.
The origin of bipedalism and its consolidation as the primary locomotor system in humans has been subject of study in recent decades thanks to the increasing discovery and analysis of the fossil record corresponding to the different taxa that made up the human lineage. Anatomical comparison with the great apes that coexist with us today allow us to identify the characteristics that differentiate orthograde bipedalism from quadrupedal locomotion and pronograde posture. Thus, the widening and shortening of the pelvis, the development of lumbar lordosis, and the modification of the foot to a purely support structure are anatomical changes resulting from a evolutionary process subject to a combination of multiple selective pressures and localized in open environments with a significant presence of trees. The fossil record shows the course of this evolution from the appearance of the first signs of facultative bipedalism in genera such as Orrorin and Ardipithecus. The genus Australopithecus presents a mosaic of arboreal and bipedal traits, with habitual bipedalism, and represents a turning point in the mode of locomotion of the human lineage. Finally, bipedal locomotion is now considered obligatory in the genus Homo, as the characteristics suggesting arboreal behavior become minimal. However, the morphology of this genus is also marked by strong sexual dimorphism resulting from the evolutionary force of obstetric difficulties caused by the expansion of the braincase and the narrowing of the birth canal.
Direction
RODRIGUEZ LUIS, JAVIER (Tutorships)
RODRIGUEZ LUIS, JAVIER (Tutorships)
Court
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
Water recovery and optimization in the wood industry
Authorship
A.G.B.
Biotechnology Degree (2nd Ed. )
A.G.B.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
The scarcity of fresh water and environmental demands have driven the wood industry to look for more sustainable processes. This Final Degree Project was carried out at the Fibranor plant (Finsa group) and focused on studying the feasibility of recovering water in the gas scrubbing system (WESP) during the production of MDF boards. Mass and energy balances were carried out in three scenarios (current situation, gas cooling and condensation recovery), supported by psychrometric analysis and characterization of water samples from the system. The samples were analyzed to evaluate their possible reuse in the process. The results indicate that it is technically possible to recover up to 7,600 L/h of water. However, its high pollutant load requires a previous treatment, such as advanced oxidation, to be able to reuse it. This strategy would reduce water consumption and improve the plant's sustainability.
The scarcity of fresh water and environmental demands have driven the wood industry to look for more sustainable processes. This Final Degree Project was carried out at the Fibranor plant (Finsa group) and focused on studying the feasibility of recovering water in the gas scrubbing system (WESP) during the production of MDF boards. Mass and energy balances were carried out in three scenarios (current situation, gas cooling and condensation recovery), supported by psychrometric analysis and characterization of water samples from the system. The samples were analyzed to evaluate their possible reuse in the process. The results indicate that it is technically possible to recover up to 7,600 L/h of water. However, its high pollutant load requires a previous treatment, such as advanced oxidation, to be able to reuse it. This strategy would reduce water consumption and improve the plant's sustainability.
Direction
BALBOA MENDEZ, SABELA (Tutorships)
BALBOA MENDEZ, SABELA (Tutorships)
Court
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
Evaluation of the biostimulant activity of Ulva lactuca extracts on different plant models.
Authorship
V.G.A.
Bachelor of Biology
V.G.A.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
In recent years, seaweed extracts have been used as potential biostimulants to enhance plant development in various species of interest, aiming to improve crop productivity and quality while reducing the environmental impact of synthetic chemical fertilizers on soil and water resources. In this study, the effect of Ulva lactuca extracts on growth parameters was evaluated in Vitis vinifera cv. Albariño seedlings grown in vitro and Arabidopsis thaliana seedlings grown in vivo. The results revealed a negative effect on the development of axillary buds, stem length, and root length in grapevine seedlings when using seaweed extract concentrations of 1% or 2.2%, with or without cyclodextrin. This effect was potentially attributed to their abscisic acid (ABA) and/or NaCl content. However, no significant impact of the extracts on total phenolic compounds in leaves was observed. On the other hand, foliar application of the extracts at a concentration of 5.4 L/ha on A. thaliana seedlings appeared to increase the number of leaves formed, though no significant differences were found compared to untreated plants after 30 days of growth. Finally, a toxicity assay was conducted on the alternative model organism Caenorhabditis elegans, confirming the safe use of the extracts. In conclusion, the biostimulant effect of seaweed extracts depends on numerous and diverse factors, such as concentration and application method, which complicate their optimal use. Therefore, further trials considering all these variables are necessary to adjust their characteristics to specific cultivation processes.
In recent years, seaweed extracts have been used as potential biostimulants to enhance plant development in various species of interest, aiming to improve crop productivity and quality while reducing the environmental impact of synthetic chemical fertilizers on soil and water resources. In this study, the effect of Ulva lactuca extracts on growth parameters was evaluated in Vitis vinifera cv. Albariño seedlings grown in vitro and Arabidopsis thaliana seedlings grown in vivo. The results revealed a negative effect on the development of axillary buds, stem length, and root length in grapevine seedlings when using seaweed extract concentrations of 1% or 2.2%, with or without cyclodextrin. This effect was potentially attributed to their abscisic acid (ABA) and/or NaCl content. However, no significant impact of the extracts on total phenolic compounds in leaves was observed. On the other hand, foliar application of the extracts at a concentration of 5.4 L/ha on A. thaliana seedlings appeared to increase the number of leaves formed, though no significant differences were found compared to untreated plants after 30 days of growth. Finally, a toxicity assay was conducted on the alternative model organism Caenorhabditis elegans, confirming the safe use of the extracts. In conclusion, the biostimulant effect of seaweed extracts depends on numerous and diverse factors, such as concentration and application method, which complicate their optimal use. Therefore, further trials considering all these variables are necessary to adjust their characteristics to specific cultivation processes.
Direction
GONZALEZ GONZALEZ, MARIA VICTORIA (Tutorships)
GONZALEZ GONZALEZ, MARIA VICTORIA (Tutorships)
Court
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
Effect of GSK-3 inhibitors and temperature on somatic embryogenesis of Picea abies
Authorship
M.G.R.
Double bachelor degree in Chemistry and Biology
M.G.R.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Picea abies (Norway spruce) is a conifer of great ecological and economic importance. One option to meet the increasing demand is somatic embryogenesis, a biotechnological technique that enables the clonal propagation of elite genotypes. Futhermore, it allows for the production of individuals adapted to local cultivation conditions through the induction of an epigenetic memory to temperature, which shapes phenotypic traits. For this reason, it’s also a pioneering technique for developing plants adapted to new climate scenarios. The potential of this technique is limited by the loss of embryogenic capacity after several proliferation cycles, making it necessary to explore alternatives to maintain the production of individuals. Therefore, in this study, GSK-3 inhibitors were used with the aim of increasing the efficiency and yield of somatic embryos. The main objective of this work is to evaluate the effect of using GSK-3 inhibitors (TDZD-8 and VP3-15) on the induction and maturation of somatic embryos at different temperatures (18, 23 and 28ºC). To determinate the effect of the inhibitors, GSK-3 enzymatic activity was quantified, and macroscopic observations were conducted to assess somatic embryo production. The results showed a direct relationship between temperature and enzymatic activity, suggesting that GSK-3 activity increases in response to rising temperature. On the other hand, the inhibitors had no effect on embryo production. This is attributed to the fact that inhibitory molecules are genotype-dependent, indicating that the clones used were nor suitable for this study. This pioneering study in this species provides new insights into the effect of GSK-3 inhibitors in relation to temperature and their potential efficiency as mediators in the production of somatic embryos in Picea abies.
Picea abies (Norway spruce) is a conifer of great ecological and economic importance. One option to meet the increasing demand is somatic embryogenesis, a biotechnological technique that enables the clonal propagation of elite genotypes. Futhermore, it allows for the production of individuals adapted to local cultivation conditions through the induction of an epigenetic memory to temperature, which shapes phenotypic traits. For this reason, it’s also a pioneering technique for developing plants adapted to new climate scenarios. The potential of this technique is limited by the loss of embryogenic capacity after several proliferation cycles, making it necessary to explore alternatives to maintain the production of individuals. Therefore, in this study, GSK-3 inhibitors were used with the aim of increasing the efficiency and yield of somatic embryos. The main objective of this work is to evaluate the effect of using GSK-3 inhibitors (TDZD-8 and VP3-15) on the induction and maturation of somatic embryos at different temperatures (18, 23 and 28ºC). To determinate the effect of the inhibitors, GSK-3 enzymatic activity was quantified, and macroscopic observations were conducted to assess somatic embryo production. The results showed a direct relationship between temperature and enzymatic activity, suggesting that GSK-3 activity increases in response to rising temperature. On the other hand, the inhibitors had no effect on embryo production. This is attributed to the fact that inhibitory molecules are genotype-dependent, indicating that the clones used were nor suitable for this study. This pioneering study in this species provides new insights into the effect of GSK-3 inhibitors in relation to temperature and their potential efficiency as mediators in the production of somatic embryos in Picea abies.
Direction
VIEJO SOMOANO, MARCOS (Tutorships)
VIEJO SOMOANO, MARCOS (Tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Experimental Study and Modeling of Protonation Mechanisms in Mosses
Authorship
M.G.R.
Double bachelor degree in Chemistry and Biology
M.G.R.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Mosses are good bioindicators of atmospheric and aquatic pollution. Their use as bioindicators is widely established because it is an inexpensive, simple, and highly reliable technique used for the detection of heavy metals. Understanding and characterizing the acid-base properties of the moss surfaces is essential for the subsequent determination of their metal retention capacity. In the present study, the protonation mechanisms of two moss species: Sphagnum palustre and Sphagnum denticulatum, are evaluated using the NICA and NICA-Donnan models. The fitting was performed based on charge curves obtained through acid-base titrations at different ionic strengths. These models allow for the determination of the distribution, accessibility, and abundance of active functional groups on the adsorption surface. It is shown that there are differences in adsorption capacity between the two mosses, with S. palustre being a more effective biosorbent. Furthermore, discontinuous titrations provide a better approach to understanding acid-base properties. The NICA-Donnan model is found to be suitable for predictive modeling, as it allows for the estimation of intrinsic parameters. Compared to other natural bioadsorbents, mosses are effective and versatile bioadsorbents, making them good biomonitors of environmental pollution.
Mosses are good bioindicators of atmospheric and aquatic pollution. Their use as bioindicators is widely established because it is an inexpensive, simple, and highly reliable technique used for the detection of heavy metals. Understanding and characterizing the acid-base properties of the moss surfaces is essential for the subsequent determination of their metal retention capacity. In the present study, the protonation mechanisms of two moss species: Sphagnum palustre and Sphagnum denticulatum, are evaluated using the NICA and NICA-Donnan models. The fitting was performed based on charge curves obtained through acid-base titrations at different ionic strengths. These models allow for the determination of the distribution, accessibility, and abundance of active functional groups on the adsorption surface. It is shown that there are differences in adsorption capacity between the two mosses, with S. palustre being a more effective biosorbent. Furthermore, discontinuous titrations provide a better approach to understanding acid-base properties. The NICA-Donnan model is found to be suitable for predictive modeling, as it allows for the estimation of intrinsic parameters. Compared to other natural bioadsorbents, mosses are effective and versatile bioadsorbents, making them good biomonitors of environmental pollution.
Direction
Fiol López, Sarah (Tutorships)
Antelo Martínez, Juan (Co-tutorships)
Fiol López, Sarah (Tutorships)
Antelo Martínez, Juan (Co-tutorships)
Court
VAZQUEZ RODRIGUEZ, SAULO ANGEL (Chairman)
BARCIELA ALONSO, Ma CARMEN (Secretary)
RIVADULLA FERNANDEZ, JOSE FRANCISCO (Member)
VAZQUEZ RODRIGUEZ, SAULO ANGEL (Chairman)
BARCIELA ALONSO, Ma CARMEN (Secretary)
RIVADULLA FERNANDEZ, JOSE FRANCISCO (Member)
Castanea sativa, Phytophthora cinnamomi, a tinta, CRISPR/Cas9, edición xenética, tolerancia, CsPMR4, xenes de susceptibilidade, embrixénese somática.
Authorship
E.G.S.
Biotechnology Degree (2nd Ed. )
E.G.S.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 09:00
07.17.2025 09:00
Summary
Castanea sativa Mill. is a highly versatile species of great ecological, economic, and cultural importance throughout Europe. However, it faces serious threats such as ink disease and chestnut blight, caused by Phytophthora cinnamomi and Cryphonectria parasitica, respectively, whose severity has increased in recent years due to climate change. The development of gene editing using CRISPR/Cas9 technology has enabled efficient and precise targeted mutagenesis. A promising gene editing strategy to confer pathogen tolerance involves inactivating susceptibility genes in plants. In this Bachelor's Degree Final Project, the silencing of the susceptibility gene CsPMR4, which encodes a callose synthase, was investigated using CRISPR/Cas9. For this purpose, somatic embryos from two embryogenic chestnut lines (CI-9 and CI-3) were transformed using Agrobacterium tumefaciens strain EHA105 carrying a CRISPR/Cas9 construct targeting the CsPMR4 gene. After ten weeks, kanamycin-resistant explants were observed only in line CI-9 (6,7 %). To evaluate editing efficiency and identify mutation types, gene amplification was performed using PCR, followed by Sanger sequencing and mutation analysis with TIDE software. Editing was confirmed in five out of the seven embryogenic lines analyzed, and four of them showed high editing efficiency (greater than 93%). To assess the tolerance of the resulting lines to the oomycete Phytophthora cinnamomi, an infection assay was conducted using somatic embryos from all isolated lines. The highest survival rates were observed in lines CI-9-PMR4-4, CI-9-PMR4-4BIS, and CI-9-PMR4-6, all with percentages above 50%.
Castanea sativa Mill. is a highly versatile species of great ecological, economic, and cultural importance throughout Europe. However, it faces serious threats such as ink disease and chestnut blight, caused by Phytophthora cinnamomi and Cryphonectria parasitica, respectively, whose severity has increased in recent years due to climate change. The development of gene editing using CRISPR/Cas9 technology has enabled efficient and precise targeted mutagenesis. A promising gene editing strategy to confer pathogen tolerance involves inactivating susceptibility genes in plants. In this Bachelor's Degree Final Project, the silencing of the susceptibility gene CsPMR4, which encodes a callose synthase, was investigated using CRISPR/Cas9. For this purpose, somatic embryos from two embryogenic chestnut lines (CI-9 and CI-3) were transformed using Agrobacterium tumefaciens strain EHA105 carrying a CRISPR/Cas9 construct targeting the CsPMR4 gene. After ten weeks, kanamycin-resistant explants were observed only in line CI-9 (6,7 %). To evaluate editing efficiency and identify mutation types, gene amplification was performed using PCR, followed by Sanger sequencing and mutation analysis with TIDE software. Editing was confirmed in five out of the seven embryogenic lines analyzed, and four of them showed high editing efficiency (greater than 93%). To assess the tolerance of the resulting lines to the oomycete Phytophthora cinnamomi, an infection assay was conducted using somatic embryos from all isolated lines. The highest survival rates were observed in lines CI-9-PMR4-4, CI-9-PMR4-4BIS, and CI-9-PMR4-6, all with percentages above 50%.
Direction
SAMPEDRO JIMÉNEZ, JAVIER (Tutorships)
Corredoira Castro, María Elena (Co-tutorships)
SAMPEDRO JIMÉNEZ, JAVIER (Tutorships)
Corredoira Castro, María Elena (Co-tutorships)
Court
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Parasites of marine mammals
Authorship
S.G.V.
Bachelor of Biology
S.G.V.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Marine mammals are considered sentinel species of aquatic ecosystems. Numerous studies have been conducted in this environment, yet parasitic infections have not been a central focus, despite their clinical, ecological, and zoonotic importance. This thesis arises from the motivation to demonstrate that parasites are significant and underestimated threats. The main objective is to identify the principal groups of parasites and examine their transmission, effects, and associated risks. Based on this, a literature review was carried out using the One Health approach. The results highlight the most common parasitic groups (protozoa, helminths, and ectoparasites) affecting marine mammals, following an updated review of scientific literature. Transmission pathways, consequences for the hosts, and impacts on public health are described. The conclusions indicate that pollution, climate change, and human activity facilitate their spread in the ecosystem, revealing the importance of applying the One Health framework. This Bachelor's Thesis aims to raise awareness about an under-researched issue and suggests expanding investigation in this highly relevant and interesting field.
Marine mammals are considered sentinel species of aquatic ecosystems. Numerous studies have been conducted in this environment, yet parasitic infections have not been a central focus, despite their clinical, ecological, and zoonotic importance. This thesis arises from the motivation to demonstrate that parasites are significant and underestimated threats. The main objective is to identify the principal groups of parasites and examine their transmission, effects, and associated risks. Based on this, a literature review was carried out using the One Health approach. The results highlight the most common parasitic groups (protozoa, helminths, and ectoparasites) affecting marine mammals, following an updated review of scientific literature. Transmission pathways, consequences for the hosts, and impacts on public health are described. The conclusions indicate that pollution, climate change, and human activity facilitate their spread in the ecosystem, revealing the importance of applying the One Health framework. This Bachelor's Thesis aims to raise awareness about an under-researched issue and suggests expanding investigation in this highly relevant and interesting field.
Direction
IGLESIAS PIÑEIRO, FRANCISCO JAVIER (Tutorships)
IGLESIAS PIÑEIRO, FRANCISCO JAVIER (Tutorships)
Court
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
Discovery and use of bacterial enzymes for plastic degradation
Authorship
E.H.C.
Biotechnology Degree (2nd Ed. )
E.H.C.
Biotechnology Degree (2nd Ed. )
Defense date
07.16.2025 10:30
07.16.2025 10:30
Summary
Plastic pollution is one of the most pressing environmental challenges worldwide. In this context, biodegradation by microbial enzymes emerges as a sustainable alternative while also promoting the transition toward a circular economy. Plastics exhibit structural diversity (bonds, crystallinity, etc.) that influences their biodegradation potential. Only a small fraction of them are fully biodegradable, whereas most are recalcitrant (lacking enzymatically accessible bonds) or hydrolyzable (containing ester bonds that are accessible but shielded by aromatic groups). Although recycling strategies and known enzymes capable of degrading them (such as DuraPETase and LCC-ICCG) exist, their effectiveness remains limited against certain polymers, particularly polyether polyurethane and high molecular weight polyamides. As several studies have shown that bacteria present in sewage sludge can metabolically adapt to use plastics as a carbon source, it is hypothesized that these microorganisms may harbor enzymes capable of degrading such complex polymers. Therefore, the present study aimed to identify a novel polyamidase enzyme in bacterial genomes isolated from wastewater treatment plant (WWTP) sludge. In addition, the study involved the production of the enzyme DuraPETase and an analysis of the degradative capacity of the recombinant cutinase LCC-ICCG against various commercial plastics. To achieve this, genomic DNA from these bacteria was extracted, sequenced, annotated, compared, and analyzed. Furthermore, the DuraPETase protein was purified, and degradation assays were conducted using the LCC-ICCG enzyme and plastics such as polyester, polyurethane, high-crystallinity PET (HC-PET), and low-crystallinity PET (LC-PET). Although the gene encoding the candidate enzyme could not be amplified, the assay results indicate degradative activity against these materials, which aligns with and reinforces previous studies conducted with this enzyme.
Plastic pollution is one of the most pressing environmental challenges worldwide. In this context, biodegradation by microbial enzymes emerges as a sustainable alternative while also promoting the transition toward a circular economy. Plastics exhibit structural diversity (bonds, crystallinity, etc.) that influences their biodegradation potential. Only a small fraction of them are fully biodegradable, whereas most are recalcitrant (lacking enzymatically accessible bonds) or hydrolyzable (containing ester bonds that are accessible but shielded by aromatic groups). Although recycling strategies and known enzymes capable of degrading them (such as DuraPETase and LCC-ICCG) exist, their effectiveness remains limited against certain polymers, particularly polyether polyurethane and high molecular weight polyamides. As several studies have shown that bacteria present in sewage sludge can metabolically adapt to use plastics as a carbon source, it is hypothesized that these microorganisms may harbor enzymes capable of degrading such complex polymers. Therefore, the present study aimed to identify a novel polyamidase enzyme in bacterial genomes isolated from wastewater treatment plant (WWTP) sludge. In addition, the study involved the production of the enzyme DuraPETase and an analysis of the degradative capacity of the recombinant cutinase LCC-ICCG against various commercial plastics. To achieve this, genomic DNA from these bacteria was extracted, sequenced, annotated, compared, and analyzed. Furthermore, the DuraPETase protein was purified, and degradation assays were conducted using the LCC-ICCG enzyme and plastics such as polyester, polyurethane, high-crystallinity PET (HC-PET), and low-crystallinity PET (LC-PET). Although the gene encoding the candidate enzyme could not be amplified, the assay results indicate degradative activity against these materials, which aligns with and reinforces previous studies conducted with this enzyme.
Direction
BALBOA MENDEZ, SABELA (Tutorships)
EIBES GONZALEZ, GEMMA MARIA (Co-tutorships)
BALBOA MENDEZ, SABELA (Tutorships)
EIBES GONZALEZ, GEMMA MARIA (Co-tutorships)
Court
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
Analysis of the Biological and Functional Traits (BTA) of Free-Living Marine Nematodes in Intertidal Soft Substrates of Galicia.
Authorship
J.H.D.
Double bachelor degree in Chemistry and Biology
J.H.D.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
The analysis of biological traits (BTA) is a tool that allows the study of nematode communities from a functional point of view, as it considers both the combinations of traits expressed by the animals and the spatial distribution of the taxa. The traits commonly studied are: adult shape, adult length, buccal morphology, tail shape, and life cycle strategy. This analysis was applied to the 25 species and 73 genera of nematodes obtained from 17 intertidal localities along the Galician coast, with feeding type also added. No significant differences were found between the taxonomic composition and the biological traits of the communities with respect to sedimentary parameters. The communities are dominated by epistrate feeders or non-selective deposit feeders, scraping microvores, with conical or clavate tails, slender, with lengths ranging from 1 to 4 mm and a life strategy of extreme-intermediate colonizer (cp 2 and 3). The samples show significant differences between exposed and sheltered localities, with transitional positions for semi-exposed environments, with wave exposure thus appearing to be the factor regulating their distribution. The combination of multiple traits, rather than each individual trait, is what allows more robust detection of ecological patterns, highlighting the quality of the information provided by BTA analyses.
The analysis of biological traits (BTA) is a tool that allows the study of nematode communities from a functional point of view, as it considers both the combinations of traits expressed by the animals and the spatial distribution of the taxa. The traits commonly studied are: adult shape, adult length, buccal morphology, tail shape, and life cycle strategy. This analysis was applied to the 25 species and 73 genera of nematodes obtained from 17 intertidal localities along the Galician coast, with feeding type also added. No significant differences were found between the taxonomic composition and the biological traits of the communities with respect to sedimentary parameters. The communities are dominated by epistrate feeders or non-selective deposit feeders, scraping microvores, with conical or clavate tails, slender, with lengths ranging from 1 to 4 mm and a life strategy of extreme-intermediate colonizer (cp 2 and 3). The samples show significant differences between exposed and sheltered localities, with transitional positions for semi-exposed environments, with wave exposure thus appearing to be the factor regulating their distribution. The combination of multiple traits, rather than each individual trait, is what allows more robust detection of ecological patterns, highlighting the quality of the information provided by BTA analyses.
Direction
BESTEIRO RODRIGUEZ, MARIA CELIA (Tutorships)
CARREIRA FLORES, DIEGO (Co-tutorships)
BESTEIRO RODRIGUEZ, MARIA CELIA (Tutorships)
CARREIRA FLORES, DIEGO (Co-tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Study of the Removal Efficiency of Microcontaminants in Wastewater Using Advanced Treatments
Authorship
J.H.D.
Double bachelor degree in Chemistry and Biology
J.H.D.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
The presence of emerging microcontaminants in urban wastewater poses an environmental and health challenge, as many of these compounds are not efectively removed by conventional wastewater treatment processes. In this context, advanced oxidation processes (AOPs) offer a promising solution for their elimination, with ozonation being one of the most widely used methods. This study evaluates the effectiveness of ozone, both alone and in combination with hydrogen peroxide (H2O2), in removing various microcontaminants from the effluent of a wastewater treatment plant (WWTP). To this end, experiments were conducted under four different conditions, with the H2O2-O3 ratio varying from 0 to 0.75. Samples were analyzed by solid-phase extraction (SPE) and liquid chromatography coupled with mass spectrometry (HPLC MS). The removal results show that certain compounds such as diclofenac and carbamazepine are removed efficiently by ozone alone, while others require more reactive conditions; the test at an H2O2-O3 ratio of 0.75 was found to be the most effective. The response strongly depends on the chemical structure of the contaminant, highlighting the need to adapt the treatment to the composition of the effluent and the nature of the microcontaminants. This study demonstrates that advanced ozonation can be an effective tool for meeting new regulatory targets in wastewater treatment, when operating conditions are optimized according to the contaminant profile present.
The presence of emerging microcontaminants in urban wastewater poses an environmental and health challenge, as many of these compounds are not efectively removed by conventional wastewater treatment processes. In this context, advanced oxidation processes (AOPs) offer a promising solution for their elimination, with ozonation being one of the most widely used methods. This study evaluates the effectiveness of ozone, both alone and in combination with hydrogen peroxide (H2O2), in removing various microcontaminants from the effluent of a wastewater treatment plant (WWTP). To this end, experiments were conducted under four different conditions, with the H2O2-O3 ratio varying from 0 to 0.75. Samples were analyzed by solid-phase extraction (SPE) and liquid chromatography coupled with mass spectrometry (HPLC MS). The removal results show that certain compounds such as diclofenac and carbamazepine are removed efficiently by ozone alone, while others require more reactive conditions; the test at an H2O2-O3 ratio of 0.75 was found to be the most effective. The response strongly depends on the chemical structure of the contaminant, highlighting the need to adapt the treatment to the composition of the effluent and the nature of the microcontaminants. This study demonstrates that advanced ozonation can be an effective tool for meeting new regulatory targets in wastewater treatment, when operating conditions are optimized according to the contaminant profile present.
Direction
MONTES GOYANES, ROSA MARIA (Tutorships)
RODIL RODRIGUEZ, MARIA DEL ROSARIO (Co-tutorships)
MONTES GOYANES, ROSA MARIA (Tutorships)
RODIL RODRIGUEZ, MARIA DEL ROSARIO (Co-tutorships)
Court
FERNANDEZ RODRIGUEZ, BERTA (Chairman)
ESTEVEZ VALCARCEL, CARLOS MANUEL (Secretary)
YEBRA BIURRUN, MARIA DEL CARMEN (Member)
FERNANDEZ RODRIGUEZ, BERTA (Chairman)
ESTEVEZ VALCARCEL, CARLOS MANUEL (Secretary)
YEBRA BIURRUN, MARIA DEL CARMEN (Member)
Studies on internalisation of peptide helicates in cells
Authorship
P.I.G.
Double bachelor degree in Chemistry and Biology
P.I.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
The copper helicate designed by our research group, and which will be the focus of this work, is part of a family of chemical systems that exhibit interesting DNA-binding properties, which justifies their consideration as potential therapeutic agents in the field of biomedical research. These are coordination compounds formed by a polypyridyl peptide ligand and two metal cations, Fe(II), Co(II), or in this case, Cu(II), that show selective recognition ability for three-way DNA junctions over other DNA structures, which appear transiently in the genome and, in many cases, are associated with various pathologies, such as cancer. However, their potential therapeutic application is limited by the inability to internalize into mammalian cells, as they cannot cross the membrane autonomously and, consequently, do not reach the nucleus, where the interaction with the three-way junctions or 3WJs occurs. Currently, our group is seeking new pathways for cellular internalization to replace digitonin, a non-ionic detergent used in preliminary assays, capable of permeabilizing the membrane but causing significant disruption to cellular physiology. Specifically, this project will focus on studying liposomes as an alternative pathway, given their structural and compositional analogy to the membrane. The main objective is to open the door, in the future, to the use of these compounds in biomedical research as a potential treatment for various diseases.
The copper helicate designed by our research group, and which will be the focus of this work, is part of a family of chemical systems that exhibit interesting DNA-binding properties, which justifies their consideration as potential therapeutic agents in the field of biomedical research. These are coordination compounds formed by a polypyridyl peptide ligand and two metal cations, Fe(II), Co(II), or in this case, Cu(II), that show selective recognition ability for three-way DNA junctions over other DNA structures, which appear transiently in the genome and, in many cases, are associated with various pathologies, such as cancer. However, their potential therapeutic application is limited by the inability to internalize into mammalian cells, as they cannot cross the membrane autonomously and, consequently, do not reach the nucleus, where the interaction with the three-way junctions or 3WJs occurs. Currently, our group is seeking new pathways for cellular internalization to replace digitonin, a non-ionic detergent used in preliminary assays, capable of permeabilizing the membrane but causing significant disruption to cellular physiology. Specifically, this project will focus on studying liposomes as an alternative pathway, given their structural and compositional analogy to the membrane. The main objective is to open the door, in the future, to the use of these compounds in biomedical research as a potential treatment for various diseases.
Direction
MARTINEZ COSTAS, JOSE MANUEL (Tutorships)
BARREIRO PIÑEIRO, NATALIA (Co-tutorships)
MARTINEZ COSTAS, JOSE MANUEL (Tutorships)
BARREIRO PIÑEIRO, NATALIA (Co-tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Synthesis of new functionalized peptide helicates
Authorship
P.I.G.
Double bachelor degree in Chemistry and Biology
P.I.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
The peptide helicates are chemical systems that exhibit interesting DNA-binding properties, justifying their consideration as potential therapeutic agents in biomedical research. They are coordination compounds composed of a polypyridyl peptide ligand and two metal cations, such as Cu(II), Co(II), or Fe(II), with the ability to selectively recognize three-way DNA junctions over other DNA structures. These junctions can transiently appear in the genome and are associated with pathologies, such as cancer. The aim of this TFG is to search for a solution, with a view to potential therapeutic application, to the main limitation of these metallopeptides: their inability to internalize cells, as they cannot cross the membrane autonomously. Currently, our group is exploring alternative strategies for the cellular internalization of these compounds to replace digitonin, which has been used effectively in preliminary assays but causes significant disruption to cellular physiology. Specifically, we will focus on the use of systems known as Cell Penetrating Peptides (CPPs), small peptides that induce the translocation of other compounds into the cell interior. This will be approached through the functionalization of the precursor peptide ligand of the helicates with a CPP composed of eight arginine residues. We will assess whether this strategy allows peptide helicates to efficiently penetrate the cells and reach the nucleus, while maintaining the selectivity of their non-functionalised analogues through three-way DNA junctions.
The peptide helicates are chemical systems that exhibit interesting DNA-binding properties, justifying their consideration as potential therapeutic agents in biomedical research. They are coordination compounds composed of a polypyridyl peptide ligand and two metal cations, such as Cu(II), Co(II), or Fe(II), with the ability to selectively recognize three-way DNA junctions over other DNA structures. These junctions can transiently appear in the genome and are associated with pathologies, such as cancer. The aim of this TFG is to search for a solution, with a view to potential therapeutic application, to the main limitation of these metallopeptides: their inability to internalize cells, as they cannot cross the membrane autonomously. Currently, our group is exploring alternative strategies for the cellular internalization of these compounds to replace digitonin, which has been used effectively in preliminary assays but causes significant disruption to cellular physiology. Specifically, we will focus on the use of systems known as Cell Penetrating Peptides (CPPs), small peptides that induce the translocation of other compounds into the cell interior. This will be approached through the functionalization of the precursor peptide ligand of the helicates with a CPP composed of eight arginine residues. We will assess whether this strategy allows peptide helicates to efficiently penetrate the cells and reach the nucleus, while maintaining the selectivity of their non-functionalised analogues through three-way DNA junctions.
Direction
VAZQUEZ LOPEZ, MIGUEL (Tutorships)
Alvar Gil, David (Co-tutorships)
VAZQUEZ LOPEZ, MIGUEL (Tutorships)
Alvar Gil, David (Co-tutorships)
Court
SAA RODRIGUEZ, CARLOS EUGENIO (Chairman)
SOUSA PEDRARES, ANTONIO (Secretary)
GARCIA DEIBE, ANA MARIA (Member)
SAA RODRIGUEZ, CARLOS EUGENIO (Chairman)
SOUSA PEDRARES, ANTONIO (Secretary)
GARCIA DEIBE, ANA MARIA (Member)
Evaluation of the effect of the ketone body beta-hydroxybutyrate on the methylome of human breast cancer cell lines.
Authorship
L.I.R.
Biotechnology Degree (2nd Ed. )
L.I.R.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
Obesity is a chronic, complex, and multifactorial disease that has become a global public health problem and is associated with an increased risk of developing various comorbidities, including cancer. The most widely used method for diagnosis is the Body Mass Index (BMI), although more precise techniques exist to assess body composition. Among therapeutic strategies, the very-low-calorie ketogenic diet (VLCKD) has played a prominent role, promoting weight loss at the expense of fat mass and inducing a state of nutritional ketosis. Furthermore, obesity generates an inflammatory microenvironment and oxidative stress, caused by dysfunctional adipose tissue, as well as epigenetic alterations. These alterations have been proposed as possible mechanistic links between this condition and its associated diseases, such as cancer. It is hypothesized that ketone bodies could favorably interfere with tumor development by regulating epigenetic mechanisms, especially DNA methylation. Therefore, the main objective of this study was to evaluate in vitro how ketone bodies, specifically beta-hydroxybutyrate, affect the methylome of human breast cancer cell lines MCF-7 and MDA-MB-231. For this study, a bioinformatic and statistical analysis of the changes in the methylome of DNA samples derived from the treated breast cancer cell lines was performed. These samples were hybridized on the MethylationEPIC BeadChip Infinium (Illumina). 292,180 differentially methylated CpG sites were identified in MDA-MB-231 and 153,458 in MCF-7, with distinct epigenetic profiles between the two cell lines, indicating that ketone bodies can modulate DNA methylation in a cell subtype-specific manner, reinforcing the idea that they could play a beneficial effect on tumor development.
Obesity is a chronic, complex, and multifactorial disease that has become a global public health problem and is associated with an increased risk of developing various comorbidities, including cancer. The most widely used method for diagnosis is the Body Mass Index (BMI), although more precise techniques exist to assess body composition. Among therapeutic strategies, the very-low-calorie ketogenic diet (VLCKD) has played a prominent role, promoting weight loss at the expense of fat mass and inducing a state of nutritional ketosis. Furthermore, obesity generates an inflammatory microenvironment and oxidative stress, caused by dysfunctional adipose tissue, as well as epigenetic alterations. These alterations have been proposed as possible mechanistic links between this condition and its associated diseases, such as cancer. It is hypothesized that ketone bodies could favorably interfere with tumor development by regulating epigenetic mechanisms, especially DNA methylation. Therefore, the main objective of this study was to evaluate in vitro how ketone bodies, specifically beta-hydroxybutyrate, affect the methylome of human breast cancer cell lines MCF-7 and MDA-MB-231. For this study, a bioinformatic and statistical analysis of the changes in the methylome of DNA samples derived from the treated breast cancer cell lines was performed. These samples were hybridized on the MethylationEPIC BeadChip Infinium (Illumina). 292,180 differentially methylated CpG sites were identified in MDA-MB-231 and 153,458 in MCF-7, with distinct epigenetic profiles between the two cell lines, indicating that ketone bodies can modulate DNA methylation in a cell subtype-specific manner, reinforcing the idea that they could play a beneficial effect on tumor development.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
CRUJEIRAS MARTINEZ, ANA BELEN (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
CRUJEIRAS MARTINEZ, ANA BELEN (Co-tutorships)
Court
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
Study of the replication of a recombinant Betanodavirus strain in sole and turbot
Authorship
M.J.R.
Bachelor of Biology
M.J.R.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Aquaculture fish production is vulnerable due to the spread of aquatic pathogens, such as the nervous necrosis virus (genus Betanodavirus). This single-stranded RNA virus is classified into 4 genotypes whose distribution depends on the temperature of the water: Barfin Flounder Nervous Necrosis Virus (BFNNV), Redspotted Grouper Nervous Necrosis Virus (RGNNV), Striped Jack Nervous Necrosis Virus (SJNNV), and Tiger Puffer Nervous Necrosis Virus (TPNNV). Recombinant strains have been described in southern Europe (RGNNV/SJNNV and SJNNV/RGNNV). In this work, the pathogenicity of a recombinant RGNNV/SJNNV isolate, obtained from mackerel, was evaluated in juvenile sole and turbot. The results showed that the Ssc1002.21 strain did not cause mortality or replicated effectively in sole. The genomic copy number was low and constant throughout the trial, and the infective viral particles were below the detection limit of the titration method. However, a significant increase in the number of genomic copies was observed in turbot samples, and viral titration in cell culture revealed the existence of a high number of infectious particles. These differences could be due to mutations in the Ssc1002.21 strain in the polymerase and in the capsid protein, located in key regions for replication and interaction with the host cell. The results highlight the importance of determining the pathogenic potential of Betanodavirus strains isolated from the wild environment for cultured species, as viruses present in wildlife represent a threat to farming systems.
Aquaculture fish production is vulnerable due to the spread of aquatic pathogens, such as the nervous necrosis virus (genus Betanodavirus). This single-stranded RNA virus is classified into 4 genotypes whose distribution depends on the temperature of the water: Barfin Flounder Nervous Necrosis Virus (BFNNV), Redspotted Grouper Nervous Necrosis Virus (RGNNV), Striped Jack Nervous Necrosis Virus (SJNNV), and Tiger Puffer Nervous Necrosis Virus (TPNNV). Recombinant strains have been described in southern Europe (RGNNV/SJNNV and SJNNV/RGNNV). In this work, the pathogenicity of a recombinant RGNNV/SJNNV isolate, obtained from mackerel, was evaluated in juvenile sole and turbot. The results showed that the Ssc1002.21 strain did not cause mortality or replicated effectively in sole. The genomic copy number was low and constant throughout the trial, and the infective viral particles were below the detection limit of the titration method. However, a significant increase in the number of genomic copies was observed in turbot samples, and viral titration in cell culture revealed the existence of a high number of infectious particles. These differences could be due to mutations in the Ssc1002.21 strain in the polymerase and in the capsid protein, located in key regions for replication and interaction with the host cell. The results highlight the importance of determining the pathogenic potential of Betanodavirus strains isolated from the wild environment for cultured species, as viruses present in wildlife represent a threat to farming systems.
Direction
BANDIN MATOS, MARIA ISABEL (Tutorships)
VAZQUEZ SALGADO, LUCIA (Co-tutorships)
BANDIN MATOS, MARIA ISABEL (Tutorships)
VAZQUEZ SALGADO, LUCIA (Co-tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Effect of amphetamines on the human microbiome
Authorship
L.L.R.
Double bachelor degree in Chemistry and Biology
L.L.R.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
In recent years, the human microbiome has been the subject of much research due to its involvement in the immune system, metabolism and molecular activity. From birth, their composition varies according to various factors. This paper will focus on the impact of drugs, in particular amphetamine-type stimulants (ATS), as they are among the most abused substances worldwide. In this context, a literature review will be conducted to examine the available experimental evidence on how methamphetamine (the most common ATS) alters the composition of the microbiome, and how this dysbiosis affects human health, contributing to the development of various diseases. Methamphetamine acts as an indirect monoamine agonist, causing effects such as wakefulness, euphoria, arousal, and increased attention. However, its consumption has serious health consequences, including alterations in the microbiome. Various studies analysed show that the consumption of methamphetamine modifies the intestinal microbiota, the oral microbiota and the supragingival dental plaque microbiota, which is associated with pathologies such as obesity, intestinal inflammation, arterial hypertension, Parkinson’s disease, Alzheimer’s and certain types of cancer. All these diseases are linked to the increase or decrease of certain bacterial phyla. Therefore, it is concluded that methamphetamine consumption induces dysbiosis in microbiotas mentioned. These findings also highlight the microbiome fundamental role in the development of serious but common diseases in today’s population.
In recent years, the human microbiome has been the subject of much research due to its involvement in the immune system, metabolism and molecular activity. From birth, their composition varies according to various factors. This paper will focus on the impact of drugs, in particular amphetamine-type stimulants (ATS), as they are among the most abused substances worldwide. In this context, a literature review will be conducted to examine the available experimental evidence on how methamphetamine (the most common ATS) alters the composition of the microbiome, and how this dysbiosis affects human health, contributing to the development of various diseases. Methamphetamine acts as an indirect monoamine agonist, causing effects such as wakefulness, euphoria, arousal, and increased attention. However, its consumption has serious health consequences, including alterations in the microbiome. Various studies analysed show that the consumption of methamphetamine modifies the intestinal microbiota, the oral microbiota and the supragingival dental plaque microbiota, which is associated with pathologies such as obesity, intestinal inflammation, arterial hypertension, Parkinson’s disease, Alzheimer’s and certain types of cancer. All these diseases are linked to the increase or decrease of certain bacterial phyla. Therefore, it is concluded that methamphetamine consumption induces dysbiosis in microbiotas mentioned. These findings also highlight the microbiome fundamental role in the development of serious but common diseases in today’s population.
Direction
LEMOS RAMOS, MANUEL LUIS (Tutorships)
LEMOS RAMOS, MANUEL LUIS (Tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Use of single drop microextraction for the determination of amphetamines in urine by gas chromatography-mass spectrometry
Authorship
L.L.R.
Double bachelor degree in Chemistry and Biology
L.L.R.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
Amphetamines are a group of drugs that act as central nervous system stimulants and whose short-term consumption produces various health effects including increased heart rate and blood pressure, reduced feeling of fatigue, euphoria, decreased appetite, etc. However, long-term use can lead to dependence, anxiety, aggression, cardiovascular problems, etc. The most commonly used amphetamines include amphetamine, methamphetamine, MDMA, MDA and MDEA. In recent years, there has been a large increase in the consumption of these substances worldwide, which poses a serious problem both at individual level and for society as a whole and may lead to overdose or even death. According to this, there is a need to develop methods for their determination in biological samples. Following the principles of green chemistry, an extraction method has been proposed which is included in the so-called microextraction techniques. Therefore, the objective of this work is to optimize and validate an analytical technique for microextraction, single drop microextraction, which allows the determination and quantification of the amphetamines studied in urine samples, using gas chromatography coupled to mass spectrometry (GC-MS) as detection technique.
Amphetamines are a group of drugs that act as central nervous system stimulants and whose short-term consumption produces various health effects including increased heart rate and blood pressure, reduced feeling of fatigue, euphoria, decreased appetite, etc. However, long-term use can lead to dependence, anxiety, aggression, cardiovascular problems, etc. The most commonly used amphetamines include amphetamine, methamphetamine, MDMA, MDA and MDEA. In recent years, there has been a large increase in the consumption of these substances worldwide, which poses a serious problem both at individual level and for society as a whole and may lead to overdose or even death. According to this, there is a need to develop methods for their determination in biological samples. Following the principles of green chemistry, an extraction method has been proposed which is included in the so-called microextraction techniques. Therefore, the objective of this work is to optimize and validate an analytical technique for microextraction, single drop microextraction, which allows the determination and quantification of the amphetamines studied in urine samples, using gas chromatography coupled to mass spectrometry (GC-MS) as detection technique.
Direction
MOREDA PIÑEIRO, ANTONIO (Tutorships)
CABARCOS FERNANDEZ, PAMELA (Co-tutorships)
MOREDA PIÑEIRO, ANTONIO (Tutorships)
CABARCOS FERNANDEZ, PAMELA (Co-tutorships)
Court
FERNANDEZ RAMOS, ANTONIO (Chairman)
CASAIS LAIÑO, Mª DEL CARMEN (Secretary)
RODIL RODRIGUEZ, MARIA DEL ROSARIO (Member)
FERNANDEZ RAMOS, ANTONIO (Chairman)
CASAIS LAIÑO, Mª DEL CARMEN (Secretary)
RODIL RODRIGUEZ, MARIA DEL ROSARIO (Member)
Study of the Biology of the invasive alga Rugulopteryx okamurae in Galicia
Authorship
C.L.M.
Bachelor of Biology
C.L.M.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Rugulopteryx okamurae is a brown alga native to Southeast Asia, first detected in Europe in 2002, in the Thau Lagoon (France), where it did not exhibit invasive behavior. In 2016, it was found on the Spanish coasts, specifically in the Strait of Gibraltar, where it is considered an invasive species with significant impacts. The main motivation for this study is its recent introduction (2019) in Galicia, where it shows a high capacity for growth and dispersion, altering marine ecosystems. The primary objective of this work is to study the morphology and phenology of R. okamurae on the Galician coasts, as well as the photosynthetic capacity of stranded individuals, in order to determine their viability. To achieve this, individuals were collected in different areas and at different times, and their morphological characteristics and photosynthetic activity were studied. The known morphotypes do not show the same seasonal distribution as in other studied areas. The thick morphotype is characterized by a greater thallus width and a higher number of cell layers at the tip of the apical dichotomy and is typical of winter, while the thin morphotype presents a narrower thallus and fewer cell layers, being characteristic of summer. Additionally, there is an intermediate morphotype between the two that appears throughout the year. It has been confirmed that stranded individuals have little to no recovery capacity and, according to our results, show very severe damage.
Rugulopteryx okamurae is a brown alga native to Southeast Asia, first detected in Europe in 2002, in the Thau Lagoon (France), where it did not exhibit invasive behavior. In 2016, it was found on the Spanish coasts, specifically in the Strait of Gibraltar, where it is considered an invasive species with significant impacts. The main motivation for this study is its recent introduction (2019) in Galicia, where it shows a high capacity for growth and dispersion, altering marine ecosystems. The primary objective of this work is to study the morphology and phenology of R. okamurae on the Galician coasts, as well as the photosynthetic capacity of stranded individuals, in order to determine their viability. To achieve this, individuals were collected in different areas and at different times, and their morphological characteristics and photosynthetic activity were studied. The known morphotypes do not show the same seasonal distribution as in other studied areas. The thick morphotype is characterized by a greater thallus width and a higher number of cell layers at the tip of the apical dichotomy and is typical of winter, while the thin morphotype presents a narrower thallus and fewer cell layers, being characteristic of summer. Additionally, there is an intermediate morphotype between the two that appears throughout the year. It has been confirmed that stranded individuals have little to no recovery capacity and, according to our results, show very severe damage.
Direction
DIAZ TAPIA, PILAR (Tutorships)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Co-tutorships)
DIAZ TAPIA, PILAR (Tutorships)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Co-tutorships)
Court
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
Use of FTIR-ATR spectroscopy to characterize the composition of bone remains from heritage-interest archaeological sites in the Iberian Peninsula
Authorship
R.L.C.
Bachelor of Biology
R.L.C.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The human skeleton is composed of bone tissue that undergoes continuous change throughout life. After death, these changes persist due to interactions with the surrounding environment through diagenetic processes. This study investigates the composition and alterations experienced by the bones and their mineral component (bioapatite) using ATR-FTIR (Attenuated Total Reflectance -Fourier Transform Infrared) spectroscopy. The skeletal remains, over 3,000 years old, were recovered from different areas of the site. Statistical analyses reveal the relationships between various indices derived from the spectroscopy results, as well as the differences among the three distinct locations within the site. The bones from the most recent structure appear to be the most altered and are the only ones lacking secondary calcite. The findings indicate that the degree of bone diagenesis is more strongly influenced by the geochemical characteristics of the burial sediment than by the duration of burial. The results also suggest that bone composition is affected by ante-mortem factors such as age and sex.
The human skeleton is composed of bone tissue that undergoes continuous change throughout life. After death, these changes persist due to interactions with the surrounding environment through diagenetic processes. This study investigates the composition and alterations experienced by the bones and their mineral component (bioapatite) using ATR-FTIR (Attenuated Total Reflectance -Fourier Transform Infrared) spectroscopy. The skeletal remains, over 3,000 years old, were recovered from different areas of the site. Statistical analyses reveal the relationships between various indices derived from the spectroscopy results, as well as the differences among the three distinct locations within the site. The bones from the most recent structure appear to be the most altered and are the only ones lacking secondary calcite. The findings indicate that the degree of bone diagenesis is more strongly influenced by the geochemical characteristics of the burial sediment than by the duration of burial. The results also suggest that bone composition is affected by ante-mortem factors such as age and sex.
Direction
MARTINEZ CORTIZAS, ANTONIO MANUEL (Tutorships)
LOPEZ COSTAS, OLALLA (Co-tutorships)
MARTINEZ CORTIZAS, ANTONIO MANUEL (Tutorships)
LOPEZ COSTAS, OLALLA (Co-tutorships)
Court
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
Study of the effect of AMPK modification in GLAST astrocytes on circadian rhythms
Authorship
H.L.G.
Bachelor of Biology
H.L.G.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The circadian system regulates key physiological processes through a complex molecular clock that synchronizes internal rhythms with environmental cues such as light and feeding. Astrocytes, traditionally overshadowed by neurons, have emerged as essential elements in the integration of metabolic and circadian signals. This study focuses on AMP-activated protein kinase (AMPK), an energy sensor present in astrocytes, and its role in circadian regulation through the study of female mice with astrocyte-specific deletion of the gamma-2 subunit (Prkag2), known to induce constitutive AMPK activation. The study also aims to detect potential sex dimorphisms. Both behavioral circadian parameters and locomotor activity under light/dark cycles and constant darkness were evaluated, as well as key molecular markers in the hypothalamus. Unlike what was observed in males, females lacking the gamma-2 subunit did not display alterations in their circadian behavioral phenotype. However, at the molecular level, a loss of oscillation in the phosphorylation of AMPK and its downstream target pACC was observed, along with alterations in the expression and phosphorylation rhythms of PER2, a core component of the molecular clock. These findings confirm the involvement of AMPK in modulating the circadian clock in astrocytes and reveal a marked sexual dimorphism: females maintain stable circadian activity despite molecular deregulation, suggesting the presence of sex-specific compensatory mechanisms. Understanding these mechanisms could provide new therapeutic perspectives for circadian and metabolic disorders, reinforcing the functional relevance of astrocytes and AMPK as potential targets in biomedical research.
The circadian system regulates key physiological processes through a complex molecular clock that synchronizes internal rhythms with environmental cues such as light and feeding. Astrocytes, traditionally overshadowed by neurons, have emerged as essential elements in the integration of metabolic and circadian signals. This study focuses on AMP-activated protein kinase (AMPK), an energy sensor present in astrocytes, and its role in circadian regulation through the study of female mice with astrocyte-specific deletion of the gamma-2 subunit (Prkag2), known to induce constitutive AMPK activation. The study also aims to detect potential sex dimorphisms. Both behavioral circadian parameters and locomotor activity under light/dark cycles and constant darkness were evaluated, as well as key molecular markers in the hypothalamus. Unlike what was observed in males, females lacking the gamma-2 subunit did not display alterations in their circadian behavioral phenotype. However, at the molecular level, a loss of oscillation in the phosphorylation of AMPK and its downstream target pACC was observed, along with alterations in the expression and phosphorylation rhythms of PER2, a core component of the molecular clock. These findings confirm the involvement of AMPK in modulating the circadian clock in astrocytes and reveal a marked sexual dimorphism: females maintain stable circadian activity despite molecular deregulation, suggesting the presence of sex-specific compensatory mechanisms. Understanding these mechanisms could provide new therapeutic perspectives for circadian and metabolic disorders, reinforcing the functional relevance of astrocytes and AMPK as potential targets in biomedical research.
Direction
BARCA MAYO, OLGA (Tutorships)
LUENGO MATEOS, MARIA (Co-tutorships)
BARCA MAYO, OLGA (Tutorships)
LUENGO MATEOS, MARIA (Co-tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Reproductive behaviour in relation to forest fires of Hedychium gardnerianum Sheppard ex Ker Gawl. in two phases of invasion: introduction and establishment.
Authorship
P.L.G.
Bachelor of Biology
P.L.G.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Hedychium gardnerianum Sheppard ex Ker Gawl is one of the 100 most damaging invasive alien species in the world, with a high capacity to spread by seeds and rhizomes. Forest fires, increasingly frequent due to climate change, create favourable conditions for its expansion. This study proposes that fire does not affect the reproductive behaviour of this species, both in areas where it is in the introduction phase (such as Galicia) and in the establishment phase (such as the Azores). To test this hypothesis, populations from both sites were analysed and compared, studying the effect of different fire factors (thermal shocks, smoke, ash and charcoal) on seeds and vegetative growth. The characterisation of the populations revealed a delay in fruiting in the Galician population. Seed viability declines drastically in late spring. High temperatures completely inhibit germination and large amounts of ash slightly reduce germination. The population in the establishment phase germinates faster than that in the introduction phase. Some seeds show dormancy. Asexual reproduction is not affected by heat shock.
Hedychium gardnerianum Sheppard ex Ker Gawl is one of the 100 most damaging invasive alien species in the world, with a high capacity to spread by seeds and rhizomes. Forest fires, increasingly frequent due to climate change, create favourable conditions for its expansion. This study proposes that fire does not affect the reproductive behaviour of this species, both in areas where it is in the introduction phase (such as Galicia) and in the establishment phase (such as the Azores). To test this hypothesis, populations from both sites were analysed and compared, studying the effect of different fire factors (thermal shocks, smoke, ash and charcoal) on seeds and vegetative growth. The characterisation of the populations revealed a delay in fruiting in the Galician population. Seed viability declines drastically in late spring. High temperatures completely inhibit germination and large amounts of ash slightly reduce germination. The population in the establishment phase germinates faster than that in the introduction phase. Some seeds show dormancy. Asexual reproduction is not affected by heat shock.
Direction
REYES FERREIRA, OTILIA (Tutorships)
Cruz de la Fuente, Óscar (Co-tutorships)
REYES FERREIRA, OTILIA (Tutorships)
Cruz de la Fuente, Óscar (Co-tutorships)
Court
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
Study of the rooting capacity of axillary shoots of holm oak established from plants selected for their tolerance to Phytophthora cinnamomi Rands.
Authorship
E.L.L.
Bachelor of Biology
E.L.L.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Quercus ilex L. is the most abundant forest species on the Iberian Peninsula. In recent years, this species has been severely affected by oak decline syndrome, primarily caused by the oomycete Phytophthora cinnamomi Rands (Pc). Within the framework of the National Breeding Program for the Quercus genus against oak decline, promoted by the Ministry for the Ecological Transition and the Demographic Challenge (MITECO), holm oak individuals tolerant to Pc have been selected, and these genotypes were cloned in vitro through axillary bud proliferation. However, Q. ilex is a recalcitrant species to in vitro culture, which hinders both root formation and plant acclimatization to ex vitro conditions. Therefore, the present study focused on optimizing an efficient rooting protocol using the selected genotypes previously established in vitro. To this end, several factors influencing root induction were evaluated: (i) the composition of the mineral medium; (ii) the concentration and exposure time to indole-3-butyric acid (IBA); (iii) the concentration of the carbon source; (iv) the type of ethylene inhibitor (silver thiosulfate or silver nitrate); and (v) the addition of 5-azacytidine. The highest rooting percentages were obtained when shoots were cultured in MS medium with either full-strength macronutrients or with nitrates reduced by half, supplemented with 25 mg/L IBA applied for 48 hours, followed by transfer to an auxin-free medium containing silver thiosulfate. The addition of 5-azacytidine to the expression medium had no positive effect on rooting frequencies. Finally, during the acclimatization phase, two types of substrate were tested, with compressed peat pellets (jiffies) yielding the highest survival rates.
Quercus ilex L. is the most abundant forest species on the Iberian Peninsula. In recent years, this species has been severely affected by oak decline syndrome, primarily caused by the oomycete Phytophthora cinnamomi Rands (Pc). Within the framework of the National Breeding Program for the Quercus genus against oak decline, promoted by the Ministry for the Ecological Transition and the Demographic Challenge (MITECO), holm oak individuals tolerant to Pc have been selected, and these genotypes were cloned in vitro through axillary bud proliferation. However, Q. ilex is a recalcitrant species to in vitro culture, which hinders both root formation and plant acclimatization to ex vitro conditions. Therefore, the present study focused on optimizing an efficient rooting protocol using the selected genotypes previously established in vitro. To this end, several factors influencing root induction were evaluated: (i) the composition of the mineral medium; (ii) the concentration and exposure time to indole-3-butyric acid (IBA); (iii) the concentration of the carbon source; (iv) the type of ethylene inhibitor (silver thiosulfate or silver nitrate); and (v) the addition of 5-azacytidine. The highest rooting percentages were obtained when shoots were cultured in MS medium with either full-strength macronutrients or with nitrates reduced by half, supplemented with 25 mg/L IBA applied for 48 hours, followed by transfer to an auxin-free medium containing silver thiosulfate. The addition of 5-azacytidine to the expression medium had no positive effect on rooting frequencies. Finally, during the acclimatization phase, two types of substrate were tested, with compressed peat pellets (jiffies) yielding the highest survival rates.
Direction
GONZALEZ GONZALEZ, MARIA VICTORIA (Tutorships)
Corredoira Castro, Elena (Co-tutorships)
GONZALEZ GONZALEZ, MARIA VICTORIA (Tutorships)
Corredoira Castro, Elena (Co-tutorships)
Court
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
Characterization of atmospheric levels of particulate matter in Santiago de Compostela
Authorship
P.M.M.
Bachelor of Biology
P.M.M.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Air pollution in cities has increased significantly since the Industrial Revolution, primarily due to vehicle traffic and other human activities. Particulate matter is one of the most concerning pollutants due to its impact on respiratory and cardiovascular health, as well as its negative effects on urban vegetation. The hypothesis of this study suggests the existence of significant spatial and seasonal patterns in particle concentrations in Santiago de Compostela, influenced by meteorological conditions and the urban environment. The objectives were to characterize the spatial distribution of particulate matter, detect seasonal variations, and verify compliance with current regulations in Spain. To this end, a portable particle meter was used, allowing high-resolution sampling at 236 points in the city, both in winter and spring. The data obtained were analyzed using statistical and geostatistical tools. The results show higher concentrations in winter, with some occasional exceedances of the legal limit established for particles smaller than 10 micrometers. Significant seasonal variability was observed, with statistically significant differences between seasons, with particle concentrations being lowest and most homogeneous in spring. Furthermore, a clear spatial dependence was identified during winter, especially in areas with high traffic density or closed urban structures. These discoveries demonstrate the usefulness of portable sampling for capturing local variations in air quality. Their applications can be key to developing more effective public policies for air quality management and reducing public health risks.
Air pollution in cities has increased significantly since the Industrial Revolution, primarily due to vehicle traffic and other human activities. Particulate matter is one of the most concerning pollutants due to its impact on respiratory and cardiovascular health, as well as its negative effects on urban vegetation. The hypothesis of this study suggests the existence of significant spatial and seasonal patterns in particle concentrations in Santiago de Compostela, influenced by meteorological conditions and the urban environment. The objectives were to characterize the spatial distribution of particulate matter, detect seasonal variations, and verify compliance with current regulations in Spain. To this end, a portable particle meter was used, allowing high-resolution sampling at 236 points in the city, both in winter and spring. The data obtained were analyzed using statistical and geostatistical tools. The results show higher concentrations in winter, with some occasional exceedances of the legal limit established for particles smaller than 10 micrometers. Significant seasonal variability was observed, with statistically significant differences between seasons, with particle concentrations being lowest and most homogeneous in spring. Furthermore, a clear spatial dependence was identified during winter, especially in areas with high traffic density or closed urban structures. These discoveries demonstrate the usefulness of portable sampling for capturing local variations in air quality. Their applications can be key to developing more effective public policies for air quality management and reducing public health risks.
Direction
FERNANDEZ ESCRIBANO, JOSE ANGEL (Tutorships)
Pacín Salvador, María do Carme (Co-tutorships)
FERNANDEZ ESCRIBANO, JOSE ANGEL (Tutorships)
Pacín Salvador, María do Carme (Co-tutorships)
Court
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
AIRA RODRÍGUEZ, Mª JESÚS (Chairman)
GOMEZ RODRIGUEZ, CAROLA (Secretary)
DOMINGUEZ CONDE, JESUS (Member)
Study of the impact of Rbm3 on circadian rhythms
Authorship
P.M.P.
Bachelor of Biology
P.M.P.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Introduction: Circadian rhythms are intrinsic and self-sustained biological oscillations generated within the cells of our organism. These oscillations synchronize with the environment thanks to the central pacemaker located in the hypothalamus: the suprachiasmatic nucleus (SCN). Until recently, the SCN was considered insensitive to ambient temperature. However, recent in vitro studies indicate that some molecular pathways regulated by clock genes do respond to thermal changes. In this context, the protein “RNA Binding Motif 3” (RBM3) stands out. This work proposes that RBM3 acts as a circadian thermal sensor in the SCN and analyzes its role in the synchronization of circadian rhythms with ambient temperature in vivo. Objectives: To generate and validate a model of RBM3 silencing in the SCN and analyze the effects on metabolic cycles and body temperature. Methodology: Animal models were used in which silencing of the RBM3 protein in the SCN was induced by stereotaxic injections of viral vectors (Rbm3Sh models). The efficacy of gene silencing was validated by immunofluorescence in brain slices. The effect of temperature on circadian parameters was analyzed using GraphPad Prism after recording body temperature with thermal sensors and metabolic cycles and activity using an indirect calorimetry system. Results: Immunofluorescence revealed a 29% reduction in RBM3 expression in mutant mice, sufficient to produce detectable phenotypic effects. Additionally, a functional role of RBM3 in regulating the period of metabolic cycles in response to ambient temperature was identified. In control mice, elevated temperatures shortened the circadian period of body temperature and metabolic cycles, while low temperatures lengthened them. These changes were not observed in Rbm3Sh mice, indicating that RBM3 is necessary for the adaptation of the circadian rhythm to thermal variations. Conclusions: Our results confirm that RBM3 actively participates in modulating the circadian period of metabolic and thermal parameters according to ambient temperature. Its deficiency reduces the circadian system’s ability to synchronize with the thermal environment, highlighting its relevance as a molecular mediator in circadian adaptation to environmental changes.
Introduction: Circadian rhythms are intrinsic and self-sustained biological oscillations generated within the cells of our organism. These oscillations synchronize with the environment thanks to the central pacemaker located in the hypothalamus: the suprachiasmatic nucleus (SCN). Until recently, the SCN was considered insensitive to ambient temperature. However, recent in vitro studies indicate that some molecular pathways regulated by clock genes do respond to thermal changes. In this context, the protein “RNA Binding Motif 3” (RBM3) stands out. This work proposes that RBM3 acts as a circadian thermal sensor in the SCN and analyzes its role in the synchronization of circadian rhythms with ambient temperature in vivo. Objectives: To generate and validate a model of RBM3 silencing in the SCN and analyze the effects on metabolic cycles and body temperature. Methodology: Animal models were used in which silencing of the RBM3 protein in the SCN was induced by stereotaxic injections of viral vectors (Rbm3Sh models). The efficacy of gene silencing was validated by immunofluorescence in brain slices. The effect of temperature on circadian parameters was analyzed using GraphPad Prism after recording body temperature with thermal sensors and metabolic cycles and activity using an indirect calorimetry system. Results: Immunofluorescence revealed a 29% reduction in RBM3 expression in mutant mice, sufficient to produce detectable phenotypic effects. Additionally, a functional role of RBM3 in regulating the period of metabolic cycles in response to ambient temperature was identified. In control mice, elevated temperatures shortened the circadian period of body temperature and metabolic cycles, while low temperatures lengthened them. These changes were not observed in Rbm3Sh mice, indicating that RBM3 is necessary for the adaptation of the circadian rhythm to thermal variations. Conclusions: Our results confirm that RBM3 actively participates in modulating the circadian period of metabolic and thermal parameters according to ambient temperature. Its deficiency reduces the circadian system’s ability to synchronize with the thermal environment, highlighting its relevance as a molecular mediator in circadian adaptation to environmental changes.
Direction
BARCA MAYO, OLGA (Tutorships)
BARCA MAYO, OLGA (Tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Structure, synthesis, and transport of catechol-type siderophores in bacteria of the genus Vibrio
Authorship
M.O.M.F.
Double bachelor degree in Chemistry and Biology
M.O.M.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
Iron is a fundamental element for the growth and development of the vast majority of living organisms, and bacteria are no exception. Since it is typically found in its oxidized form (Fe3+), it is difficult to absorb directly due to its low solubility under physiological conditions. To overcome this limitation, bacteria have developed a series of iron assimilation mechanisms. One of the most important is the synthesis of low-molecular-weight compounds known as siderophores, which enable them to acquire iron from the environment. This work focuses specifically on catechol-type siderophores produced by bacteria of the genus Vibrio. These siderophores have a general structure based on 2,3-dihydroxybenzoic acid (2,3-DHBA) and norspermidine, to which different amino acids and functional groups are attached depending on the specific siderophore. Moreover, their synthesis relies on chorismate, NRPS enzymes, and the Fur regulatory system. Finally, transport is carried out by TonB-dependent outer membrane transporter proteins and ABC-type systems for translocation across the cytoplasmic membrane.
Iron is a fundamental element for the growth and development of the vast majority of living organisms, and bacteria are no exception. Since it is typically found in its oxidized form (Fe3+), it is difficult to absorb directly due to its low solubility under physiological conditions. To overcome this limitation, bacteria have developed a series of iron assimilation mechanisms. One of the most important is the synthesis of low-molecular-weight compounds known as siderophores, which enable them to acquire iron from the environment. This work focuses specifically on catechol-type siderophores produced by bacteria of the genus Vibrio. These siderophores have a general structure based on 2,3-dihydroxybenzoic acid (2,3-DHBA) and norspermidine, to which different amino acids and functional groups are attached depending on the specific siderophore. Moreover, their synthesis relies on chorismate, NRPS enzymes, and the Fur regulatory system. Finally, transport is carried out by TonB-dependent outer membrane transporter proteins and ABC-type systems for translocation across the cytoplasmic membrane.
Direction
LEMOS RAMOS, MANUEL LUIS (Tutorships)
BALADO DACOSTA, MIGUEL (Co-tutorships)
LEMOS RAMOS, MANUEL LUIS (Tutorships)
BALADO DACOSTA, MIGUEL (Co-tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Lanthanide complexes as anion sensors: A literature review
Authorship
M.O.M.F.
Double bachelor degree in Chemistry and Biology
M.O.M.F.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
In recent decades, the selective and precise detection of anions in the environment has gained significant importance due to its impact on both human health and natural ecosystems. Among the strategies studied for this purpose, the emerging use of lanthanide complexes stands out, as their unique photophysical properties allow the construction of selective sensors with fast response times and optimal spatial resolution. This work is based on a bibliographic review focused on the search for lanthanide complexes as molecular sensors or probes for the specific detection of anions in the environment, analyzing the optical signaling mechanisms involved and discussing relevant examples from recent literature. The goal is to assess their potential as effective analytical tools for environmental monitoring.
In recent decades, the selective and precise detection of anions in the environment has gained significant importance due to its impact on both human health and natural ecosystems. Among the strategies studied for this purpose, the emerging use of lanthanide complexes stands out, as their unique photophysical properties allow the construction of selective sensors with fast response times and optimal spatial resolution. This work is based on a bibliographic review focused on the search for lanthanide complexes as molecular sensors or probes for the specific detection of anions in the environment, analyzing the optical signaling mechanisms involved and discussing relevant examples from recent literature. The goal is to assess their potential as effective analytical tools for environmental monitoring.
Direction
FONDO BUSTO, MARIA MATILDE (Tutorships)
FONDO BUSTO, MARIA MATILDE (Tutorships)
Court
Estévez Cabanas, Juan Carlos (Chairman)
GARCIA SUAREZ, LUIS ALBERTO (Secretary)
GONZALEZ NOYA, ANA MARIA (Member)
Estévez Cabanas, Juan Carlos (Chairman)
GARCIA SUAREZ, LUIS ALBERTO (Secretary)
GONZALEZ NOYA, ANA MARIA (Member)
Effect of mepolizumab on the low-abundance serum proteome of patients with severe eosinophilic asthma.
Authorship
A.M.G.
Bachelor of Biology
A.M.G.
Bachelor of Biology
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Severe eosinophilic asthma (SEA) represents a subtype of the T2high phenotype characterized by persistent eosinophil-mediated inflammation and resistance to conventional treatment with inhaled corticosteroids. Interleukin 5 (IL-5) plays a key role in the activation, maturation and survival of eosinophils, making it a therapeutic target for biological drugs such as mepolizumab (an anti-IL5 antibody). This study evaluates the effect of mepolizumab treatment on the low-abundance serum proteome in patients with SEA, with samples collected from healthy controls and asthmatic patients at four time points: before treatment (T0) and at 4, 16, and 32 weeks after treatment initiation (T4, T16, T32). Samples were treated with DTT to remove the high-abundance proteome, and using mass spectrometry (LC-MS/MS) and the SWATH-MS technique, it was observed that healthy controls and patients at the end of treatment (T32) showed a similar profile. Similarly, the profiles before treatment (T0) and at 4 weeks (T4) were also alike. At T0, the proteins APOH and PROS1, related to coagulation, and SERPINA1 and SERPINA3, related to the acute phase response, were found to be overexpressed. These, along with SERPINA6, which was also overexpressed, showed high predictive power (AUC higher than 0.8), with the latter possibly having the greatest implication in the differences between healthy controls and T0, and between T0 and T32.
Severe eosinophilic asthma (SEA) represents a subtype of the T2high phenotype characterized by persistent eosinophil-mediated inflammation and resistance to conventional treatment with inhaled corticosteroids. Interleukin 5 (IL-5) plays a key role in the activation, maturation and survival of eosinophils, making it a therapeutic target for biological drugs such as mepolizumab (an anti-IL5 antibody). This study evaluates the effect of mepolizumab treatment on the low-abundance serum proteome in patients with SEA, with samples collected from healthy controls and asthmatic patients at four time points: before treatment (T0) and at 4, 16, and 32 weeks after treatment initiation (T4, T16, T32). Samples were treated with DTT to remove the high-abundance proteome, and using mass spectrometry (LC-MS/MS) and the SWATH-MS technique, it was observed that healthy controls and patients at the end of treatment (T32) showed a similar profile. Similarly, the profiles before treatment (T0) and at 4 weeks (T4) were also alike. At T0, the proteins APOH and PROS1, related to coagulation, and SERPINA1 and SERPINA3, related to the acute phase response, were found to be overexpressed. These, along with SERPINA6, which was also overexpressed, showed high predictive power (AUC higher than 0.8), with the latter possibly having the greatest implication in the differences between healthy controls and T0, and between T0 and T32.
Direction
NIETO FONTARIGO, JUAN JOSE (Tutorships)
NIETO FONTARIGO, JUAN JOSE (Tutorships)
Court
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
Development of cellular rejuvenation through partial chemical reprogramming
Authorship
M.M.T.
Bachelor of Biology
M.M.T.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Cellular senescence is a response to stress factors that halts the cell cycle. This damage-preventing state plays a dual role: in a physiological context, it supports tissue development and repair; however, in a pathological state, it promotes chronic inflammation and the progression of degenerative diseases. It has been demonstrated that cellular reprogramming using the transcription factors Oct3/4, Sox2, Klf4, and c-Myc (OSKM), as well as chemical reprogramming with seven compounds, can revert differentiated cells to an induced pluripotent state. Partial reprogramming using either these transcription factors or two of the chemical compounds can reverse aging-related effects without causing loss of cell identity. Based on this, the study explores the possibility of partially reprogramming mouse embryonic fibroblasts (MEFs), which were induced into senescence using X-ray radiation, using only: two transcription factors, Sox2 and Oct4 (SO); and two of the chemical compounds (2C), RepSox and Tranylcypromine (TCP). A positive correlation was observed between reduced senescence and the cultures treated with SO and 2C, with these compounds showing a senomorphic effect. This partial reprogramming represents a promising biomedical approach to rejuvenate tissues without triggering dangerous dedifferentiation and to potentially treat aging-related neurodegenerative, metabolic, or cardiovascular diseases.
Cellular senescence is a response to stress factors that halts the cell cycle. This damage-preventing state plays a dual role: in a physiological context, it supports tissue development and repair; however, in a pathological state, it promotes chronic inflammation and the progression of degenerative diseases. It has been demonstrated that cellular reprogramming using the transcription factors Oct3/4, Sox2, Klf4, and c-Myc (OSKM), as well as chemical reprogramming with seven compounds, can revert differentiated cells to an induced pluripotent state. Partial reprogramming using either these transcription factors or two of the chemical compounds can reverse aging-related effects without causing loss of cell identity. Based on this, the study explores the possibility of partially reprogramming mouse embryonic fibroblasts (MEFs), which were induced into senescence using X-ray radiation, using only: two transcription factors, Sox2 and Oct4 (SO); and two of the chemical compounds (2C), RepSox and Tranylcypromine (TCP). A positive correlation was observed between reduced senescence and the cultures treated with SO and 2C, with these compounds showing a senomorphic effect. This partial reprogramming represents a promising biomedical approach to rejuvenate tissues without triggering dangerous dedifferentiation and to potentially treat aging-related neurodegenerative, metabolic, or cardiovascular diseases.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
Court
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
Study of non-aromatic fluorescent peptides (GlowSticks)
Authorship
J.N.S.
Biotechnology Degree (2nd Ed. )
J.N.S.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 09:00
07.17.2025 09:00
Summary
Non-aromatic fluorescence is a little-studied phenomenon in which compounds that lack traditional aromatic groups are nonetheless capable of emitting light in the UV/Vis spectrum. The use of peptides with these properties as markers for microscopy could overcome many of the limitations associated with traditional organic or protein-based fluorophores, such as GFP. In this study, variants of 5UOI-W14 -a peptide known to exhibit non-aromatic fluorescence- were synthesized, purified, and analyzed using fluorescence spectroscopy, with modifications introduced at the N-terminal end to assess their potential as markers. It was found that additions at this site caused a significant loss of the compound’s fluorescent properties. Furthermore, using the same methodology, fluorescence was observed in various commercial proteins. These measurements revealed an anomalous emission band around 450 nm in pepsin, suggesting its potential use as a marker in future studies. Finally, expression assays were conducted in HELA cells using the fluorescent peptides (E4R4)4 and (E4R4)9 to evaluate their in vitro labeling capabilities and to determine whether successive additions of the E4R4 motif would enhance emission. However, the results were inconclusive due to the low concentration of the peptides in the culture.
Non-aromatic fluorescence is a little-studied phenomenon in which compounds that lack traditional aromatic groups are nonetheless capable of emitting light in the UV/Vis spectrum. The use of peptides with these properties as markers for microscopy could overcome many of the limitations associated with traditional organic or protein-based fluorophores, such as GFP. In this study, variants of 5UOI-W14 -a peptide known to exhibit non-aromatic fluorescence- were synthesized, purified, and analyzed using fluorescence spectroscopy, with modifications introduced at the N-terminal end to assess their potential as markers. It was found that additions at this site caused a significant loss of the compound’s fluorescent properties. Furthermore, using the same methodology, fluorescence was observed in various commercial proteins. These measurements revealed an anomalous emission band around 450 nm in pepsin, suggesting its potential use as a marker in future studies. Finally, expression assays were conducted in HELA cells using the fluorescent peptides (E4R4)4 and (E4R4)9 to evaluate their in vitro labeling capabilities and to determine whether successive additions of the E4R4 motif would enhance emission. However, the results were inconclusive due to the low concentration of the peptides in the culture.
Direction
VAZQUEZ SENTIS, MARCO EUGENIO (Tutorships)
VAZQUEZ SENTIS, MARCO EUGENIO (Tutorships)
Court
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Gómez Tato, Antonio M. (Chairman)
FERNÁNDEZ LORENZO, JUAN LUIS (Secretary)
NOIA GULDRÍS, MANUEL (Member)
Relationship between the gut microbiota and the development and/or prevention of major depression
Authorship
M.N.G.
Bachelor of Biology
M.N.G.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The present bachelor's thesis addresses to explore the relationship between the gut microbiota (GM) and major depression (MD), analyzing how intestinal dysbiosis influences its pathophysiology through neurobiological, inflammatory, and metabolic mechanisms. The objectives included reviewing the classical hypotheses of MD [monoaminergic, stress/hypothalamic-pituitary-adrenal axis (HPA axis), cytokines, and neuroplasticity], examining the role of microbial metabolites (short-chain fatty acids (SCFAs), tryptophan, LPS), and evaluating interventions based on GM modulation. The methodology consisted of a narrative review of preclinical and clinical studies, selecting research linking dysbiosis, inflammation, and behavioral alterations from the past 10 years. Findings on rodents exposed to chronic stress or induced dysbiosis were prioritized, as well as the effects of probiotics and diet. The main findings indicate that GM decreases levels of serotonin, dopamine, and BDNF, while increasing proinflammatory cytokines and oxidative stress. Additionally, it was observed that short-chain fatty acids, especially butyrate, strengthen the intestinal barrier, reduce neuroinflammation, and promote neurogenesis. In turn, certain probiotics (Lactobacillus, Bifidobacterium) were shown to reverse depressive behaviors in animal models by regulating neurotransmitters and modulating the HPA axis. In conclusion, the GM emerges as a key factor in MD, where strategies such as high-fiber diets, probiotics, and fecal transplantation could complement pharmacological treatments. Although preclinical evidence is robust, further human studies are needed to validate these interventions. This integrative approach opens new avenues for personalized therapies in mental health.
The present bachelor's thesis addresses to explore the relationship between the gut microbiota (GM) and major depression (MD), analyzing how intestinal dysbiosis influences its pathophysiology through neurobiological, inflammatory, and metabolic mechanisms. The objectives included reviewing the classical hypotheses of MD [monoaminergic, stress/hypothalamic-pituitary-adrenal axis (HPA axis), cytokines, and neuroplasticity], examining the role of microbial metabolites (short-chain fatty acids (SCFAs), tryptophan, LPS), and evaluating interventions based on GM modulation. The methodology consisted of a narrative review of preclinical and clinical studies, selecting research linking dysbiosis, inflammation, and behavioral alterations from the past 10 years. Findings on rodents exposed to chronic stress or induced dysbiosis were prioritized, as well as the effects of probiotics and diet. The main findings indicate that GM decreases levels of serotonin, dopamine, and BDNF, while increasing proinflammatory cytokines and oxidative stress. Additionally, it was observed that short-chain fatty acids, especially butyrate, strengthen the intestinal barrier, reduce neuroinflammation, and promote neurogenesis. In turn, certain probiotics (Lactobacillus, Bifidobacterium) were shown to reverse depressive behaviors in animal models by regulating neurotransmitters and modulating the HPA axis. In conclusion, the GM emerges as a key factor in MD, where strategies such as high-fiber diets, probiotics, and fecal transplantation could complement pharmacological treatments. Although preclinical evidence is robust, further human studies are needed to validate these interventions. This integrative approach opens new avenues for personalized therapies in mental health.
Direction
MARTIN CORA, FRANCISCO JAVIER (Tutorships)
MARTIN CORA, FRANCISCO JAVIER (Tutorships)
Court
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
Study of the Diatom Community in the Lor River: an Analysis of the Local Variability and Richness
Authorship
D.O.S.
Bachelor of Biology
D.O.S.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
This study characterizes the composition, structure, and local variability of the diatom community at different sampling points along the Lor River as it passes through Seoane do Courel (Lugo). The main objective is to identify the most diverse and representative site within this river section, which would make it the most suitable location for long-term monitoring. In addition to conducting species inventories and quantifying patterns of alpha diversity (i.e., species richness) and beta diversity (i.e., biotic dissimilarity) among the four studied sites, the study also infers water quality at each location using the presence/absence of different species as indicators of potential disturbances in environmental conditions. A study based on non-parametric estimators is also carried out to assess whether the sampling effort commonly accepted as standard for species diversity studies in freshwater epilithic communities (n = 500 valves) is sufficient for proper community characterization. Finally, a photographic catalog of the identified species with a relative abundance greater than 1% has been created to facilitate future monitoring of the community.
This study characterizes the composition, structure, and local variability of the diatom community at different sampling points along the Lor River as it passes through Seoane do Courel (Lugo). The main objective is to identify the most diverse and representative site within this river section, which would make it the most suitable location for long-term monitoring. In addition to conducting species inventories and quantifying patterns of alpha diversity (i.e., species richness) and beta diversity (i.e., biotic dissimilarity) among the four studied sites, the study also infers water quality at each location using the presence/absence of different species as indicators of potential disturbances in environmental conditions. A study based on non-parametric estimators is also carried out to assess whether the sampling effort commonly accepted as standard for species diversity studies in freshwater epilithic communities (n = 500 valves) is sufficient for proper community characterization. Finally, a photographic catalog of the identified species with a relative abundance greater than 1% has been created to facilitate future monitoring of the community.
Direction
GOMEZ RODRIGUEZ, CAROLA (Tutorships)
LEIRA CAMPOS, ANTON MANOEL (Co-tutorships)
GOMEZ RODRIGUEZ, CAROLA (Tutorships)
LEIRA CAMPOS, ANTON MANOEL (Co-tutorships)
Court
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
Evaluation of the functionality of a SUMOylation mutant of RPL23
Authorship
A.P.M.
Bachelor of Biology
A.P.M.
Bachelor of Biology
Defense date
07.16.2025 09:30
07.16.2025 09:30
Summary
Ribosomal proteins are proteins that make up the ribosomes. These proteins participate in protein synthesis and play a critical role in regulating various cellular processes, including cell growth and apoptosis. The ribosomal protein RPL23 belongs to the large 60S ribosomal subunit and, similarly to proteins RPL11 and RPL5, can act as an activator of the tumor suppressor p53. It is known that the activity of RPL11 is regulated through post-translational modifications by ubiquitin-like proteins, such as SUMOylation or NEDDylation. We know that RPL23 can be conjugated to SUMO, but how this modification affects the protein is still unknown. In this study, we aim to assess whether a mutant of RPL23 at lysine 66 has altered SUMO conjugation and how this modification affects the ribosomal protein. Our results indicate that the RPL23 mutant at lysine K66 has a reduced ability to conjugate to SUMO2. Moreover, we observed that the localization of the mutant is slightly affected. While the wild-type RPL23 protein is primarily located in the nucleolus, the localization of the RPL23-K66R mutant is more diffuse and nucleoplasmic. Additionally, we found that SUMO overexpression promotes the translocation of RPL23 from the nucleolus to the nucleoplasm. These results suggest that SUMO modulates the subcellular localization of the protein, but likely in an indirect manner, since both the overexpression and reduction of SUMOylation induce the delocalization of RPL23 from the nucleolus. We also studied whether ribosomal stress alters SUMOylation in RPL23. Our results suggest that such stress decreases SUMOylation of RPL23. Since a decrease in SUMOylation appears to promote the translocation of RPL23 to the nucleoplasm, this mechanism could favor the activation of p53. Further studies would be necessary to confirm these findings and identify other potential regulators of RPL23. Such identification could facilitate the control of its activity and the development of therapeutic strategies for diseases where RPL23 is involved.
Ribosomal proteins are proteins that make up the ribosomes. These proteins participate in protein synthesis and play a critical role in regulating various cellular processes, including cell growth and apoptosis. The ribosomal protein RPL23 belongs to the large 60S ribosomal subunit and, similarly to proteins RPL11 and RPL5, can act as an activator of the tumor suppressor p53. It is known that the activity of RPL11 is regulated through post-translational modifications by ubiquitin-like proteins, such as SUMOylation or NEDDylation. We know that RPL23 can be conjugated to SUMO, but how this modification affects the protein is still unknown. In this study, we aim to assess whether a mutant of RPL23 at lysine 66 has altered SUMO conjugation and how this modification affects the ribosomal protein. Our results indicate that the RPL23 mutant at lysine K66 has a reduced ability to conjugate to SUMO2. Moreover, we observed that the localization of the mutant is slightly affected. While the wild-type RPL23 protein is primarily located in the nucleolus, the localization of the RPL23-K66R mutant is more diffuse and nucleoplasmic. Additionally, we found that SUMO overexpression promotes the translocation of RPL23 from the nucleolus to the nucleoplasm. These results suggest that SUMO modulates the subcellular localization of the protein, but likely in an indirect manner, since both the overexpression and reduction of SUMOylation induce the delocalization of RPL23 from the nucleolus. We also studied whether ribosomal stress alters SUMOylation in RPL23. Our results suggest that such stress decreases SUMOylation of RPL23. Since a decrease in SUMOylation appears to promote the translocation of RPL23 to the nucleoplasm, this mechanism could favor the activation of p53. Further studies would be necessary to confirm these findings and identify other potential regulators of RPL23. Such identification could facilitate the control of its activity and the development of therapeutic strategies for diseases where RPL23 is involved.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
Rivas Vázquez, María del Carmen (Co-tutorships)
BLANQUER GARATE, MARIA (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
Rivas Vázquez, María del Carmen (Co-tutorships)
BLANQUER GARATE, MARIA (Co-tutorships)
Court
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
Enzymatic degradation of PET in sequential bioreactor.
Authorship
R.P.P.
Biotechnology Degree (2nd Ed. )
R.P.P.
Biotechnology Degree (2nd Ed. )
Defense date
07.16.2025 10:30
07.16.2025 10:30
Summary
Plastic is one of the most widely used materials worldwide due to its outstanding qualities such as high durability and flexibility. The problem with this material arises from its non-degradability in the environment, which leads to its accumulation and formation of contaminating particles. The most commonly used methods for the treatment of plastic waste are landfill disposal, incineration and recycling. The latter is the most interesting, since it allows the production of new plastic by substituting the use of virgin materials. Here we present polyethylene terephthalate (PET), one of the most widely used types of plastic in textiles and packaging. PET is a plastic that can be recycled biologically through the use of enzymes such as LCC-ICCG cutinase, which degrades it into its essential monomers and allows its synthesis without consuming new materials. This idea underlies the present work, in which the process of degradation of PET by means of an already known hydrolase, LCC-ICCG cutinase, both in the free state and immobilized by means of an IC-Tagging encapsulation system, is scaled up. In this work, the scale-up of PET degradation was carried out at a 50 mL bioreactor scale, studying the number of cycles that the enzyme can be reused before it exhausts its activity and the study of the optimal agitation to carry out such degradation assays. The experiments carried out showed that PET degradation is greatly reduced as the process is scaled up, reaching only 29,3% degradation of PET with the free enzyme, and 36,7% degradation of PET with the encapsulated enzyme. In addition, it was found that at high operating agitations, worse degradation efficiencies were obtained for PET with the encapsulated enzyme.
Plastic is one of the most widely used materials worldwide due to its outstanding qualities such as high durability and flexibility. The problem with this material arises from its non-degradability in the environment, which leads to its accumulation and formation of contaminating particles. The most commonly used methods for the treatment of plastic waste are landfill disposal, incineration and recycling. The latter is the most interesting, since it allows the production of new plastic by substituting the use of virgin materials. Here we present polyethylene terephthalate (PET), one of the most widely used types of plastic in textiles and packaging. PET is a plastic that can be recycled biologically through the use of enzymes such as LCC-ICCG cutinase, which degrades it into its essential monomers and allows its synthesis without consuming new materials. This idea underlies the present work, in which the process of degradation of PET by means of an already known hydrolase, LCC-ICCG cutinase, both in the free state and immobilized by means of an IC-Tagging encapsulation system, is scaled up. In this work, the scale-up of PET degradation was carried out at a 50 mL bioreactor scale, studying the number of cycles that the enzyme can be reused before it exhausts its activity and the study of the optimal agitation to carry out such degradation assays. The experiments carried out showed that PET degradation is greatly reduced as the process is scaled up, reaching only 29,3% degradation of PET with the free enzyme, and 36,7% degradation of PET with the encapsulated enzyme. In addition, it was found that at high operating agitations, worse degradation efficiencies were obtained for PET with the encapsulated enzyme.
Direction
EIBES GONZALEZ, GEMMA MARIA (Tutorships)
BALBOA MENDEZ, SABELA (Co-tutorships)
EIBES GONZALEZ, GEMMA MARIA (Tutorships)
BALBOA MENDEZ, SABELA (Co-tutorships)
Court
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
Study of the effect of the treatment with obestatin in an in vitro model of human pancreatic ductal adenocarcinoma
Authorship
S.P.C.
Bachelor of Biology
S.P.C.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Pancreatic ductal adenocarcinoma, which is the predominant type of cancer of pancreas, constitutes one of the most aggressive and lethal neoplasias known so far. It estimates that within two decades, this was the second cause of death by cancer in the world, in spite of its reduced incidence. In this context, the implication of the obestatin and the G protein associated receptor 39 (GPR39) in the regulation of the cellular proliferation, posits its capacity to block key processes associated with the tumoral development. This work has the objective to evaluate the potentiality of the obestatin/GPR39 system as a therapeutic target in an in vitro model of cancer of pancreas: the cellular line MIA PaCa-2. Once confirmed the expression of said system in the model, the molecular essays revealed that the treatment with obestatin stimulates the apoptosis, at the same time that inhibits the mitogenic activity of the pancreatic tumor cells. In parallel, the obestatin is able to induce the susceptibility to the hypoxia and favor anti-inflamatory states, two relevant events in the redesign of the tumoral microenvironment. In conclusion, these data reinforce the fundamental role of the obestatin/GPR39 system in the control of the tumoral growth of the pancreatic ductal adenocarcinoma, opening way to new therapeutic strategies.
Pancreatic ductal adenocarcinoma, which is the predominant type of cancer of pancreas, constitutes one of the most aggressive and lethal neoplasias known so far. It estimates that within two decades, this was the second cause of death by cancer in the world, in spite of its reduced incidence. In this context, the implication of the obestatin and the G protein associated receptor 39 (GPR39) in the regulation of the cellular proliferation, posits its capacity to block key processes associated with the tumoral development. This work has the objective to evaluate the potentiality of the obestatin/GPR39 system as a therapeutic target in an in vitro model of cancer of pancreas: the cellular line MIA PaCa-2. Once confirmed the expression of said system in the model, the molecular essays revealed that the treatment with obestatin stimulates the apoptosis, at the same time that inhibits the mitogenic activity of the pancreatic tumor cells. In parallel, the obestatin is able to induce the susceptibility to the hypoxia and favor anti-inflamatory states, two relevant events in the redesign of the tumoral microenvironment. In conclusion, these data reinforce the fundamental role of the obestatin/GPR39 system in the control of the tumoral growth of the pancreatic ductal adenocarcinoma, opening way to new therapeutic strategies.
Direction
POMBO RAMOS, CELIA MARIA (Tutorships)
Leal López, Saúl (Co-tutorships)
Pazos Randulfe, Yolanda (Co-tutorships)
POMBO RAMOS, CELIA MARIA (Tutorships)
Leal López, Saúl (Co-tutorships)
Pazos Randulfe, Yolanda (Co-tutorships)
Court
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
CORDERO SANTAMARIA, OSCAR JAVIER (Chairman)
ROMAUS SANJURJO, DANIEL (Secretary)
LOPEZ PEREZ, MIGUEL ANTONIO (Member)
Evaluation of the antimicrobial activity of a natural extract against microorganisms responsible for oral diseases and development of an active pharmaceutical formulation
Authorship
A.P.G.
Bachelor of Biology
A.P.G.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
Biofilms are associations of bacterial communities adhered to a solid surface and embedded in a matrix that provides them with protection and resistance. These structures, common in both biotic and abiotic environments, represent a major challenge in the treatment of oral disease, potentially leading to the loss of the teeth, bone, and supporting tissue. This study presents the design of a mouthwash based on a natural extract from Galician flora rich in polyphenols. The objective is to evaluate its ability to inhibit the growth and formation of biofilm against odontopathogenic bacteria such as Streptococcus mutans and Enterococcus faecalis, while preserving the oral microbiota. Antimicrobial activity assays were carried out using resazurin as redox indicator, which is reduced to fluorescent resorufin in the presence of viable cells. For biofilm inhibition, crystal violet was used to quantify the biomass adhered to the surface of the well. The results show greater effectiveness of the extract against S. mutans, both in inhibiting cell viability and biofilm formation. Against E. faecalis, the effect of the extract was limited, as it did not form quantifiable biofilm and no significant reduction in its viability was observed. Several preliminary formulations were designed with different proportions of the extract and various components commonly used in cosmetic industry. Finally, a formulation was obtained with good antimicrobial activity and biofilm reduction capacity, respectful of the oral microbiota and suitable for daily use as a preventive therapy.
Biofilms are associations of bacterial communities adhered to a solid surface and embedded in a matrix that provides them with protection and resistance. These structures, common in both biotic and abiotic environments, represent a major challenge in the treatment of oral disease, potentially leading to the loss of the teeth, bone, and supporting tissue. This study presents the design of a mouthwash based on a natural extract from Galician flora rich in polyphenols. The objective is to evaluate its ability to inhibit the growth and formation of biofilm against odontopathogenic bacteria such as Streptococcus mutans and Enterococcus faecalis, while preserving the oral microbiota. Antimicrobial activity assays were carried out using resazurin as redox indicator, which is reduced to fluorescent resorufin in the presence of viable cells. For biofilm inhibition, crystal violet was used to quantify the biomass adhered to the surface of the well. The results show greater effectiveness of the extract against S. mutans, both in inhibiting cell viability and biofilm formation. Against E. faecalis, the effect of the extract was limited, as it did not form quantifiable biofilm and no significant reduction in its viability was observed. Several preliminary formulations were designed with different proportions of the extract and various components commonly used in cosmetic industry. Finally, a formulation was obtained with good antimicrobial activity and biofilm reduction capacity, respectful of the oral microbiota and suitable for daily use as a preventive therapy.
Direction
Miguel Bouzas, María Trinidad de (Tutorships)
Gómez Calvo, Lorena (Co-tutorships)
Miguel Bouzas, María Trinidad de (Tutorships)
Gómez Calvo, Lorena (Co-tutorships)
Court
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
Evaluation of thermally-induced unfolding of recombinant bank vole PrPC carrying a pathogenic mutation that induces prion disease
Authorship
R.P.L.
Biotechnology Degree (2nd Ed. )
R.P.L.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 12:30
07.17.2025 12:30
Summary
Prions are infectious agents composed solely of protein and cause fatal neurodegenerative diseases. Their pathogenicity is associated with the conversion of the cellular form of the prion protein into its infectious, misfolded form, which aggregates into amyloid fibrils. Certain mutations increase the propensity to develop prion diseases. One such mutation is the substitution of glutamate with lysine at position 200 (E200K). In this context, a study will be initiated to evaluate the heat-induced unfolding of the recombinant prion protein from the bank vole carrying the isoleucine polymorphism at residue 109 and the E200K mutation. The prion protein will be expressed and purified for Nuclear Magnetic Resonance (NMR) spectroscopy and Circular Dichroism (CD) spectroscopy experiments. NMR will be used to assess the thermal unfolding process and its reversibility, while CD spectra will reveal changes in the protein’s secondary structure. The results will be compared to those obtained for the wild-type version of the bank vole prion protein. It has been found that the mutant protein behaves significantly differently from the wild-type. Although both proteins show similar thermal stability, the structural changes induced by heat unfolding are reversible only in the E200K mutant. This reversibility is nearly complete, and the observed changes after heating are likely due to an increased B-sheet content. These findings are expected to contribute to a better understanding of the conversion process from the cellular to the pathogenic isoform of the prion protein.
Prions are infectious agents composed solely of protein and cause fatal neurodegenerative diseases. Their pathogenicity is associated with the conversion of the cellular form of the prion protein into its infectious, misfolded form, which aggregates into amyloid fibrils. Certain mutations increase the propensity to develop prion diseases. One such mutation is the substitution of glutamate with lysine at position 200 (E200K). In this context, a study will be initiated to evaluate the heat-induced unfolding of the recombinant prion protein from the bank vole carrying the isoleucine polymorphism at residue 109 and the E200K mutation. The prion protein will be expressed and purified for Nuclear Magnetic Resonance (NMR) spectroscopy and Circular Dichroism (CD) spectroscopy experiments. NMR will be used to assess the thermal unfolding process and its reversibility, while CD spectra will reveal changes in the protein’s secondary structure. The results will be compared to those obtained for the wild-type version of the bank vole prion protein. It has been found that the mutant protein behaves significantly differently from the wild-type. Although both proteins show similar thermal stability, the structural changes induced by heat unfolding are reversible only in the E200K mutant. This reversibility is nearly complete, and the observed changes after heating are likely due to an increased B-sheet content. These findings are expected to contribute to a better understanding of the conversion process from the cellular to the pathogenic isoform of the prion protein.
Direction
RODRIGUEZ REQUENA, JESUS (Tutorships)
RODRIGUEZ REQUENA, JESUS (Tutorships)
Court
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
Comparative bioavailability study based on the size and composition of synthetic resins to be removed from cultural heritage.
Authorship
I.P.L.
Bachelor of Biology
I.P.L.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Different exploratory assays were carried out aiming to increase the bioavailability (accessibility to living organisms) of xenobiotics (synthetic substances that are difficult to eliminate) to bacteria, with the goal of facilitating their removal from cultural heritage through biocleaning processes (the use of living organisms and/or their enzymes as cleaning agents). Three bacterial strains were used, previously shown to have proven biodegradative capabilities as biocleaning agents on spray paint graffiti (considered a xenobiotic): Comamonas (formerly Pseudomonas) testosteroni, Enterobacter (formerly Aerobacter and currently Klebsiella) aerogenes, and Rhodococcus erythropolis. Two xenobiotics were tested: Paraloid B72 and Paraloid B82. To improve bioavailability, two liquid-medium assays were conducted. In the first, the particle size of both Paraloids was varied (diameters of 4 mm, 1 mm, and 0.25 mm were tested), under the hypothesis that smaller sizes would be more accessible to bacteria. In the second assay, pretreatments using two detergent-like additives (Tween80 and Tween20) and one organic solvent, dimethyl sulfoxide (DMSO), were applied. In both assays, the percentage of weight loss of the resins was measured, and in the surrounding liquid medium, absorbance at 600 nm (related to the presence of bacteria), redox potential, and dissolved organic (COD) and inorganic carbon (CID) were analyzed. The results showed that smaller particle sizes appear to enhance the bioavailability of both Paraloids to the bacteria, with Rhodococcus erythropolis showing particularly strong activity. Paraloid B82 may be easier to bioremove than its counterpart, despite having a very similar chemical structure. Among the three pretreatments, Tween80 appears to perform best in promoting the biocleaning process, followed by Tween20.
Different exploratory assays were carried out aiming to increase the bioavailability (accessibility to living organisms) of xenobiotics (synthetic substances that are difficult to eliminate) to bacteria, with the goal of facilitating their removal from cultural heritage through biocleaning processes (the use of living organisms and/or their enzymes as cleaning agents). Three bacterial strains were used, previously shown to have proven biodegradative capabilities as biocleaning agents on spray paint graffiti (considered a xenobiotic): Comamonas (formerly Pseudomonas) testosteroni, Enterobacter (formerly Aerobacter and currently Klebsiella) aerogenes, and Rhodococcus erythropolis. Two xenobiotics were tested: Paraloid B72 and Paraloid B82. To improve bioavailability, two liquid-medium assays were conducted. In the first, the particle size of both Paraloids was varied (diameters of 4 mm, 1 mm, and 0.25 mm were tested), under the hypothesis that smaller sizes would be more accessible to bacteria. In the second assay, pretreatments using two detergent-like additives (Tween80 and Tween20) and one organic solvent, dimethyl sulfoxide (DMSO), were applied. In both assays, the percentage of weight loss of the resins was measured, and in the surrounding liquid medium, absorbance at 600 nm (related to the presence of bacteria), redox potential, and dissolved organic (COD) and inorganic carbon (CID) were analyzed. The results showed that smaller particle sizes appear to enhance the bioavailability of both Paraloids to the bacteria, with Rhodococcus erythropolis showing particularly strong activity. Paraloid B82 may be easier to bioremove than its counterpart, despite having a very similar chemical structure. Among the three pretreatments, Tween80 appears to perform best in promoting the biocleaning process, followed by Tween20.
Direction
SANMARTIN SANCHEZ, PATRICIA (Tutorships)
Martín Caramés, Fabiana (Co-tutorships)
SANMARTIN SANCHEZ, PATRICIA (Tutorships)
Martín Caramés, Fabiana (Co-tutorships)
Court
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
LOPEZ RODRIGUEZ, Mª DEL CARMEN (Chairman)
VIEJO SOMOANO, MARCOS (Secretary)
LEIRA CAMPOS, ANTON MANOEL (Member)
Role of small extracellular vesicles (sEVs) in the energy balance of cachexia
Authorship
S.B.P.Q.
Bachelor of Biology
S.B.P.Q.
Bachelor of Biology
Defense date
07.16.2025 09:30
07.16.2025 09:30
Summary
Cancer cachexia is a multifactorial syndrome involving profound alterations in energy balance. It is characterized by significant weight loss, systemic inflammation, anorexia, and increased energy expenditure due to the activation of processes such as thermogenesis. Among the emerging therapeutic targets is AMP-activated protein kinase (AMPK), which functions as an energy sensor and a key regulator of metabolism in the hypothalamus, particularly in neurons of the ventromedial nucleus. In this context, this study aims to evaluate whether systemic administration of small extracellular vesicles (sEVs) loaded with constitutively active AMPKa1 and specifically targeting SF1 neurons in the hypothalamus can reverse the effects of C26 tumor-induced cachexia in mice. For this purpose, an animal model of cachexia generated by subcutaneous injection of C26 tumor cells, followed by systemic treatment with sEVs, was used. Parameters such as body weight, food intake, body composition, and brown adipose tissue thermogenesis were subsequently analyzed. The results showed a significant loss of body weight and fat in the tumor-bearing animals, validating the model. Furthermore, administration of sEVs failed to reverse the weight loss in the cachectic animals, but did produce a significant increase in food intake and a tendency toward reduced thermogenesis. These findings suggest that sEVs may represent a promising strategy for the treatment of cachexia by modulating hypothalamic metabolism while avoiding invasive procedures.
Cancer cachexia is a multifactorial syndrome involving profound alterations in energy balance. It is characterized by significant weight loss, systemic inflammation, anorexia, and increased energy expenditure due to the activation of processes such as thermogenesis. Among the emerging therapeutic targets is AMP-activated protein kinase (AMPK), which functions as an energy sensor and a key regulator of metabolism in the hypothalamus, particularly in neurons of the ventromedial nucleus. In this context, this study aims to evaluate whether systemic administration of small extracellular vesicles (sEVs) loaded with constitutively active AMPKa1 and specifically targeting SF1 neurons in the hypothalamus can reverse the effects of C26 tumor-induced cachexia in mice. For this purpose, an animal model of cachexia generated by subcutaneous injection of C26 tumor cells, followed by systemic treatment with sEVs, was used. Parameters such as body weight, food intake, body composition, and brown adipose tissue thermogenesis were subsequently analyzed. The results showed a significant loss of body weight and fat in the tumor-bearing animals, validating the model. Furthermore, administration of sEVs failed to reverse the weight loss in the cachectic animals, but did produce a significant increase in food intake and a tendency toward reduced thermogenesis. These findings suggest that sEVs may represent a promising strategy for the treatment of cachexia by modulating hypothalamic metabolism while avoiding invasive procedures.
Direction
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ESTEVEZ SALGUERO, ANXELA MARIA (Co-tutorships)
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ESTEVEZ SALGUERO, ANXELA MARIA (Co-tutorships)
Court
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
Regulation of PML by the Ebola Virus VP24 Protein.
Authorship
X.P.S.
Bachelor of Biology
X.P.S.
Bachelor of Biology
Defense date
07.16.2025 09:30
07.16.2025 09:30
Summary
The Ebola virus causes a hemorrhagic fever with a high case fatality rate. One of its viral proteins, VP24, interacts with the viral nucleoprotein (NP), which is essential for replication, thereby promoting the formation of the nucleocapsid. VP24 also plays a crucial role in evading the immune response by antagonizing the interferon pathway through direct interaction with signal transducers and activators of transcription, or via interaction with karyopherins. Another effect of VP24 is the alteration of the nuclear membrane through interactions with several of its components. It has been reported that damage to the nuclear membrane may lead to changes in the subcellular distribution of various cellular components, including the promyelocytic leukemia protein (PML). PML is a predominantly nuclear protein with multiple functions, among them an antiviral role, although it can also be exploited by viruses at certain stages of their replication cycle. PML is an essential constituent of PML nuclear bodies, as it recruits the various proteins that make up these macromolecular aggregates. According to preliminary laboratory results, expression of the Ebola virus VP24 protein may induce the translocation of PML from the nucleus to the cytoplasm. Whether this translocation could facilitate interactions of PML and VP24 and NP remains unknown. In this study, we address these questions. To this end, we performed immunofluorescence assays, subcellular fractionation, and co-immunoprecipitation experiments. Our results confirm that VP24 induces the cytoplasmic translocation of PML and that PML interacts with both VP24 and NP. These findings suggest that such interactions may have a positive impact on viral replication, which will be the subject of future studies.
The Ebola virus causes a hemorrhagic fever with a high case fatality rate. One of its viral proteins, VP24, interacts with the viral nucleoprotein (NP), which is essential for replication, thereby promoting the formation of the nucleocapsid. VP24 also plays a crucial role in evading the immune response by antagonizing the interferon pathway through direct interaction with signal transducers and activators of transcription, or via interaction with karyopherins. Another effect of VP24 is the alteration of the nuclear membrane through interactions with several of its components. It has been reported that damage to the nuclear membrane may lead to changes in the subcellular distribution of various cellular components, including the promyelocytic leukemia protein (PML). PML is a predominantly nuclear protein with multiple functions, among them an antiviral role, although it can also be exploited by viruses at certain stages of their replication cycle. PML is an essential constituent of PML nuclear bodies, as it recruits the various proteins that make up these macromolecular aggregates. According to preliminary laboratory results, expression of the Ebola virus VP24 protein may induce the translocation of PML from the nucleus to the cytoplasm. Whether this translocation could facilitate interactions of PML and VP24 and NP remains unknown. In this study, we address these questions. To this end, we performed immunofluorescence assays, subcellular fractionation, and co-immunoprecipitation experiments. Our results confirm that VP24 induces the cytoplasmic translocation of PML and that PML interacts with both VP24 and NP. These findings suggest that such interactions may have a positive impact on viral replication, which will be the subject of future studies.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
Rivas Vázquez, María del Carmen (Co-tutorships)
Tolosa , Rocío Miranda (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
Rivas Vázquez, María del Carmen (Co-tutorships)
Tolosa , Rocío Miranda (Co-tutorships)
Court
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
SANTOS RODRIGUEZ, MARIA ISABEL (Chairman)
DIAZ JULLIEN, CRISTINA (Secretary)
MARTIN CORA, FRANCISCO JAVIER (Member)
Identification of bacteria potentially pathogenic in the clam microbiota associated with mortality due to the increase in temperature in the water.
Authorship
M.R.D.A.L.
Bachelor of Biology
M.R.D.A.L.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
The production of bivalve molluscs in Galicia, especially carpet shell (Venerupis corrugata), has experienced a marked decline in recent years, possibly related to the effects of climate change. Increasing water temperature may cause an imbalance in the microbiota of these bivalves, promoting the proliferation of opportunistic bacteria of the genus Vibrio, responsible for the disease called vibriosis. This work aims to detect the presence of potentially pathogenic bacteria in the carpet shell microbiota after following an episode of increasing water temperature. For this, a test was carried out clams were exposed to a temperature increase of 15 ºC (control) to 20 ºC, triggering an outbreak of mortality associated with dysbiosis. There were bacteria from both healthy and affected animals, and an analysis was carried out phenotypic in relation to the presence of virulence factors: hemolysis, activity gelatinase, lipase and siderophore production. The results indicated an delete of bacterial isolates with greater frequency and intensity of these activities of clams subjected to thermal stress, evidencing a microbiota enriched in potentially pathogenic bacteria. In addition, the usefulness of detection by PCR of frpA and abtA genes, involved in the transport of siderophores piscibactina and amphibactin, as possible predictive molecular markers of mortality. In conclusión, the production of amphibactin is closely related to higher bacterial virulence, suggesting its value as an early indicator to detect infectious outbreaks.
The production of bivalve molluscs in Galicia, especially carpet shell (Venerupis corrugata), has experienced a marked decline in recent years, possibly related to the effects of climate change. Increasing water temperature may cause an imbalance in the microbiota of these bivalves, promoting the proliferation of opportunistic bacteria of the genus Vibrio, responsible for the disease called vibriosis. This work aims to detect the presence of potentially pathogenic bacteria in the carpet shell microbiota after following an episode of increasing water temperature. For this, a test was carried out clams were exposed to a temperature increase of 15 ºC (control) to 20 ºC, triggering an outbreak of mortality associated with dysbiosis. There were bacteria from both healthy and affected animals, and an analysis was carried out phenotypic in relation to the presence of virulence factors: hemolysis, activity gelatinase, lipase and siderophore production. The results indicated an delete of bacterial isolates with greater frequency and intensity of these activities of clams subjected to thermal stress, evidencing a microbiota enriched in potentially pathogenic bacteria. In addition, the usefulness of detection by PCR of frpA and abtA genes, involved in the transport of siderophores piscibactina and amphibactin, as possible predictive molecular markers of mortality. In conclusión, the production of amphibactin is closely related to higher bacterial virulence, suggesting its value as an early indicator to detect infectious outbreaks.
Direction
LEMOS RAMOS, MANUEL LUIS (Tutorships)
BALADO DACOSTA, MIGUEL (Co-tutorships)
LEMOS RAMOS, MANUEL LUIS (Tutorships)
BALADO DACOSTA, MIGUEL (Co-tutorships)
Court
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
ARES MAZAS, MARIA ELVIRA (Chairman)
BALBOA MENDEZ, SABELA (Secretary)
BARREIRO IGLESIAS, ANTON (Member)
Hypothalamic AMPK targeting for the treatment of obesity using therapeutic cellsomes.
Authorship
T.R.F.
Bachelor of Biology
T.R.F.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Nowadays, obesity is spreading globally as one of the most prevalent diseases, potentially reaching pandemic proportions in the short term. Given the existing limitations for its therapy, nanomedicine is proposed as a promising alternative for its treatment and that of its many associated pathologies. Multiple studies have shown that the loss of AMPKa1 (AMP-activated protein kinase alpha 1) function in steroidogenic factor 1 (SF1) neurons of the ventromedial nucleus of the hypothalamus causes resistance to obesity, independent of food intake due to sympathetic activation of brown adipose tissue thermogenesis. The aim of the present study is to investigate whether the body weight of obese mice can be reduced by intravenous injection of cell-derived nanocarriers, known as cellsomes, capable of crossing the blood-brain barrier and reaching specific areas of the hypothalamus (where they can exert modulatory effects on energy balance). Thus, these cells carry a plasmid (functioning as a vector) encoding a dominant-negative version of AMPKa1 (AMPKa1-DN) targeting SF1 neurons in the ventromedial nucleus of the hypothalamus. In this study, we will describe the central effect of these cellsomes in murine models of diet-induced obesity and study their involvement in the thermogenic activity of brown adipose tissue dependent on UCP1 (uncoupling protein 1). The results obtained demonstrate that the central action of cellsomes loaded with the SF1-AMPKa1-DN plasmid produces an increase in brown adipose tissue thermogenesis and consequent weight loss independent of food intake.
Nowadays, obesity is spreading globally as one of the most prevalent diseases, potentially reaching pandemic proportions in the short term. Given the existing limitations for its therapy, nanomedicine is proposed as a promising alternative for its treatment and that of its many associated pathologies. Multiple studies have shown that the loss of AMPKa1 (AMP-activated protein kinase alpha 1) function in steroidogenic factor 1 (SF1) neurons of the ventromedial nucleus of the hypothalamus causes resistance to obesity, independent of food intake due to sympathetic activation of brown adipose tissue thermogenesis. The aim of the present study is to investigate whether the body weight of obese mice can be reduced by intravenous injection of cell-derived nanocarriers, known as cellsomes, capable of crossing the blood-brain barrier and reaching specific areas of the hypothalamus (where they can exert modulatory effects on energy balance). Thus, these cells carry a plasmid (functioning as a vector) encoding a dominant-negative version of AMPKa1 (AMPKa1-DN) targeting SF1 neurons in the ventromedial nucleus of the hypothalamus. In this study, we will describe the central effect of these cellsomes in murine models of diet-induced obesity and study their involvement in the thermogenic activity of brown adipose tissue dependent on UCP1 (uncoupling protein 1). The results obtained demonstrate that the central action of cellsomes loaded with the SF1-AMPKa1-DN plasmid produces an increase in brown adipose tissue thermogenesis and consequent weight loss independent of food intake.
Direction
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ESTEVEZ SALGUERO, ANXELA MARIA (Co-tutorships)
LOPEZ PEREZ, MIGUEL ANTONIO (Tutorships)
ESTEVEZ SALGUERO, ANXELA MARIA (Co-tutorships)
Court
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
GARCIA SUAREZ, CARLOS (Chairman)
VIDAL FIGUEROA, ANXO (Secretary)
BANDIN MATOS, MARIA ISABEL (Member)
Determination of potential genetic interactions between deregulated versions of YEN1 and genes involved in the removal of DNA protein adducts
Authorship
L.R.F.
Biotechnology Degree (2nd Ed. )
L.R.F.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
The maintenance of genome stability relies on a broad set of DNA repair mechanisms, among which structure-specific nucleases play a key role. In Saccharomyces cerevisiae, the endonuclease Yen1 performs an essential function as a Holliday junction resolvase, and its activity is tightly regulated throughout the cell cycle. Several studies have shown that deregulated versions of Yen1- either constitutively expressed or phosphomutant-can be toxic if they access DNA prematurely during S phase. In this context, the expression of a catalytically inactive variant (Yen1ON-ND) retains such toxicity, suggesting the formation of protein-DNA crosslinks (DPCs), a highly obstructive type of lesion that interferes with both replication and transcription. This Bachelor’s Thesis investigates whether the overexpression of factors involved in DPC resolution can suppress such toxicity. To this end, the genes WSS1, CDC48, and DDI1 were cloned using the Gateway system and constitutively expressed in S. cerevisiae. WSS1 encodes a DNA-dependent protease that acts on DPCs in cooperation with the ATPase Cdc48, which segregates SUMO- or ubiquitin-modified proteins from chromatin. DDI1, in turn, may function redundantly when the main repair pathways fail. Although the interaction between these proteins and Yen1ON-ND could not be confirmed, the results suggest that WSS1 itself may be toxic under the conditions tested. This work lays the foundation for future studies to explore the suppressive potential of WSS1, CDC48, and DDI1 under alternative inducible promoters.
The maintenance of genome stability relies on a broad set of DNA repair mechanisms, among which structure-specific nucleases play a key role. In Saccharomyces cerevisiae, the endonuclease Yen1 performs an essential function as a Holliday junction resolvase, and its activity is tightly regulated throughout the cell cycle. Several studies have shown that deregulated versions of Yen1- either constitutively expressed or phosphomutant-can be toxic if they access DNA prematurely during S phase. In this context, the expression of a catalytically inactive variant (Yen1ON-ND) retains such toxicity, suggesting the formation of protein-DNA crosslinks (DPCs), a highly obstructive type of lesion that interferes with both replication and transcription. This Bachelor’s Thesis investigates whether the overexpression of factors involved in DPC resolution can suppress such toxicity. To this end, the genes WSS1, CDC48, and DDI1 were cloned using the Gateway system and constitutively expressed in S. cerevisiae. WSS1 encodes a DNA-dependent protease that acts on DPCs in cooperation with the ATPase Cdc48, which segregates SUMO- or ubiquitin-modified proteins from chromatin. DDI1, in turn, may function redundantly when the main repair pathways fail. Although the interaction between these proteins and Yen1ON-ND could not be confirmed, the results suggest that WSS1 itself may be toxic under the conditions tested. This work lays the foundation for future studies to explore the suppressive potential of WSS1, CDC48, and DDI1 under alternative inducible promoters.
Direction
González Blanco, Miguel (Tutorships)
González Blanco, Miguel (Tutorships)
Court
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
Development of targeted systems for apoptosis induction
Authorship
A.R.R.
Biotechnology Degree (2nd Ed. )
A.R.R.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
Intercellular communication plays a key role in the response to tissue damage. The regulation of processes such as programmed cell death and senescence, commonly considered independent mechanisms, is essential for the proper repair and regeneration of damaged tissue. In this context, recent studies in our laboratory have suggested that certain forms of cell death could induce senescence in neighboring cells through paracrine signals. This study aimed to explore this hypothesis using a mitochondria-independent, controlled apoptosis system based on the induced activation of caspase 9. To this end, a TC1 cell line modified to express an inducible version of caspase 9 (iCasp9-GFP), activated by the dimerizing compound AP20187, was generated. The apoptotic cells were exposed to primary murine embryonic fibroblasts using in vitro cultures with conditioned media, transwell systems, and direct co-culture, after which the potential induction of senescence was evaluated. In all cases, the results showed that apoptosis induction was not sufficient to paracrine-activate a senescent phenotype in the target cells, in contrast to positive controls of cells induced to senescence by treatment with traditional chemotherapeutics. These findings allow us to separate the cell death process from the accompanying signaling, suggesting that this signaling (and not apoptosis itself) is responsible for inducing paracrine senescence, with other factors such as mitochondrial dysfunction or the immunological context being possible triggers. The study provides a useful framework for further exploring the complex interplay between cell death and senescence, with potential therapeutic implications for diseases associated with aging or tissue regeneration.
Intercellular communication plays a key role in the response to tissue damage. The regulation of processes such as programmed cell death and senescence, commonly considered independent mechanisms, is essential for the proper repair and regeneration of damaged tissue. In this context, recent studies in our laboratory have suggested that certain forms of cell death could induce senescence in neighboring cells through paracrine signals. This study aimed to explore this hypothesis using a mitochondria-independent, controlled apoptosis system based on the induced activation of caspase 9. To this end, a TC1 cell line modified to express an inducible version of caspase 9 (iCasp9-GFP), activated by the dimerizing compound AP20187, was generated. The apoptotic cells were exposed to primary murine embryonic fibroblasts using in vitro cultures with conditioned media, transwell systems, and direct co-culture, after which the potential induction of senescence was evaluated. In all cases, the results showed that apoptosis induction was not sufficient to paracrine-activate a senescent phenotype in the target cells, in contrast to positive controls of cells induced to senescence by treatment with traditional chemotherapeutics. These findings allow us to separate the cell death process from the accompanying signaling, suggesting that this signaling (and not apoptosis itself) is responsible for inducing paracrine senescence, with other factors such as mitochondrial dysfunction or the immunological context being possible triggers. The study provides a useful framework for further exploring the complex interplay between cell death and senescence, with potential therapeutic implications for diseases associated with aging or tissue regeneration.
Direction
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
VIDAL FIGUEROA, ANXO (Tutorships)
Collado Rodríguez, Manuel (Co-tutorships)
DA SILVA ALVAREZ, SABELA (Co-tutorships)
Court
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
Identification of the replication origin and analysis of the distribution of plasmid pPHDP60 in Photobacterium damselae subsp. piscicida.
Authorship
C.R.C.
Biotechnology Degree (2nd Ed. )
C.R.C.
Biotechnology Degree (2nd Ed. )
Defense date
07.16.2025 10:30
07.16.2025 10:30
Summary
The replication of bacterial plasmids depends on a specific sequence known as the origin of replication (oriV), which is essential for ensuring their stability in the host cell. The plasmid pPHDP60, present in the marine fish pathogen Photobacterium damselae subsp. piscicida, lacks iterons and classical rep genes, suggesting an atypical replication mechanism. Studying the minimal oriV of pPHDP60 could have relevant applications in basic research and in the design of molecular tools for biotechnology or epidemiological surveillance. This study hypothesizes that its oriV may consist of non-conventional elements and that its distribution in P. damselae might be underestimated. The main objective was to identify its minimal functional oriV and to analyze its distribution using experimental and bioinformatic approaches. To achieve this, combined strategies of molecular cloning, transformation in Escherichia coli, bioinformatic analysis, and PCR screening were applied to a collection of P. damselae subsp. piscicida isolates. A 2,289 bp fragment (designated as IB), composed of two intergenic regions and an open reading frame for a hypothetical protein, was identified. This fragment enabled autonomous replication in E. coli, confirming its functionality as a replicon. Moreover, the evidence obtained in this study suggests that all elements of the IB sequence are required together for replication. In silico analysis showed that the IB sequence is conserved in various P. damselae strains, although variability exists in other modules that constitute the characteristic structure of pPHDP60. Additionally, one of the regions within IB (RI1) is also conserved in other plasmids of the Vibrionaceae family. Finally, PCR screening of various piscicida subsp. isolates confirmed that the presence of the pPHDP60 plasmid, in this subspecies, is restricted to gilthead seabream (Sparus aurata) isolates from Galician fish farms.
The replication of bacterial plasmids depends on a specific sequence known as the origin of replication (oriV), which is essential for ensuring their stability in the host cell. The plasmid pPHDP60, present in the marine fish pathogen Photobacterium damselae subsp. piscicida, lacks iterons and classical rep genes, suggesting an atypical replication mechanism. Studying the minimal oriV of pPHDP60 could have relevant applications in basic research and in the design of molecular tools for biotechnology or epidemiological surveillance. This study hypothesizes that its oriV may consist of non-conventional elements and that its distribution in P. damselae might be underestimated. The main objective was to identify its minimal functional oriV and to analyze its distribution using experimental and bioinformatic approaches. To achieve this, combined strategies of molecular cloning, transformation in Escherichia coli, bioinformatic analysis, and PCR screening were applied to a collection of P. damselae subsp. piscicida isolates. A 2,289 bp fragment (designated as IB), composed of two intergenic regions and an open reading frame for a hypothetical protein, was identified. This fragment enabled autonomous replication in E. coli, confirming its functionality as a replicon. Moreover, the evidence obtained in this study suggests that all elements of the IB sequence are required together for replication. In silico analysis showed that the IB sequence is conserved in various P. damselae strains, although variability exists in other modules that constitute the characteristic structure of pPHDP60. Additionally, one of the regions within IB (RI1) is also conserved in other plasmids of the Vibrionaceae family. Finally, PCR screening of various piscicida subsp. isolates confirmed that the presence of the pPHDP60 plasmid, in this subspecies, is restricted to gilthead seabream (Sparus aurata) isolates from Galician fish farms.
Direction
RODRIGUEZ OSORIO, CARLOS (Tutorships)
Vázquez Barca, Alba (Co-tutorships)
RODRIGUEZ OSORIO, CARLOS (Tutorships)
Vázquez Barca, Alba (Co-tutorships)
Court
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
Effect of osmotic stress on the maturation of somatic embryos of grapevine (Vitis vinifera L.)
Authorship
D.S.G.
Double bachelor degree in Chemistry and Biology
D.S.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 09:00
07.16.2025 09:00
Summary
Somatic embryogenesis is a very useful biotechnological tool in species such as grapevines, where genetic improvement is limited due to the need to maintain cultivar genotypes. This species is considered recalcitrant to the application of this technology, but our research group recently developed an efficient protocol for inducing somatic embryogenesis in the Galician cultivar Mencía. However, during the differentiation stage of somatic embryos, cases of abnormal development and premature germination were detected, resulting in a decreased seedling conversion rate. To optimize the maturation and differentiation process, embryos must achieve an adequate morphology and undergo the necessary physiological processes. Therefore, the objective of this study is the effect of water stress on the differentiation and maturation process of somatic embryos. To this end, embryogenic aggregates were incubated under different water stress treatments based on semi-permeable membranes or osmotic agents such as polyethylene glycol 6000 (PEG). It was observed that embryos with lower water content had fewer germinants but higher conversion rates. This suggests that reducing water availability favors proper embryo maturation, resulting in viable plants. In parallel with the study, gene expression levels of a gene essential for abscisic acid (ABA) synthesis were assayed to support the above hypothesis.
Somatic embryogenesis is a very useful biotechnological tool in species such as grapevines, where genetic improvement is limited due to the need to maintain cultivar genotypes. This species is considered recalcitrant to the application of this technology, but our research group recently developed an efficient protocol for inducing somatic embryogenesis in the Galician cultivar Mencía. However, during the differentiation stage of somatic embryos, cases of abnormal development and premature germination were detected, resulting in a decreased seedling conversion rate. To optimize the maturation and differentiation process, embryos must achieve an adequate morphology and undergo the necessary physiological processes. Therefore, the objective of this study is the effect of water stress on the differentiation and maturation process of somatic embryos. To this end, embryogenic aggregates were incubated under different water stress treatments based on semi-permeable membranes or osmotic agents such as polyethylene glycol 6000 (PEG). It was observed that embryos with lower water content had fewer germinants but higher conversion rates. This suggests that reducing water availability favors proper embryo maturation, resulting in viable plants. In parallel with the study, gene expression levels of a gene essential for abscisic acid (ABA) synthesis were assayed to support the above hypothesis.
Direction
MARTINEZ TRONCOSO, OSCAR (Tutorships)
MARTINEZ TRONCOSO, OSCAR (Tutorships)
Court
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
BARJA FRANCISCO, PRIMITIVO (Chairman)
FIDALGO PEREZ, MIGUEL ANGEL (Secretary)
VIÑAS DIAZ, ANA MARIA (Member)
Cluster helicates from silver salts
Authorship
D.S.G.
Double bachelor degree in Chemistry and Biology
D.S.G.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
The aim of this Final Degree thesis is to obtain silver(I) metallosupramolecular architectures of the cluster helicate type. For this purpose, a family of four bisthiosemicarbazone ligands that differ in the substituents of their arms has been used. The Ag(I) complexes were synthesised by traditional chemical synthesis using salts with low coordinating bulky anions (hexafluorophosphate and tetrafluoroborate) and an anion with a moderate base character (acetate). Both ligands and complexes were characterised by different techniques in solid state and in solution, including elemental analysis, IR, mass spectrometry, 1H NMR and, where possible, X-ray diffraction. Cationic tetranuclear Ag(I) complexes with hexafluorofostate and tetrafluoroborate anions have been obtained, while neutral tetranuclear cluster helicates are stabilised from acetate.
The aim of this Final Degree thesis is to obtain silver(I) metallosupramolecular architectures of the cluster helicate type. For this purpose, a family of four bisthiosemicarbazone ligands that differ in the substituents of their arms has been used. The Ag(I) complexes were synthesised by traditional chemical synthesis using salts with low coordinating bulky anions (hexafluorophosphate and tetrafluoroborate) and an anion with a moderate base character (acetate). Both ligands and complexes were characterised by different techniques in solid state and in solution, including elemental analysis, IR, mass spectrometry, 1H NMR and, where possible, X-ray diffraction. Cationic tetranuclear Ag(I) complexes with hexafluorofostate and tetrafluoroborate anions have been obtained, while neutral tetranuclear cluster helicates are stabilised from acetate.
Direction
GONZALEZ NOYA, ANA MARIA (Tutorships)
BARREIRO SISTO, UXIA (Co-tutorships)
GONZALEZ NOYA, ANA MARIA (Tutorships)
BARREIRO SISTO, UXIA (Co-tutorships)
Court
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
Evaluation of the antibacterial activity of hydrogels base don alkyl bisamides with encapsulated antibiotics
Authorship
L.S.M.
Double bachelor degree in Chemistry and Biology
L.S.M.
Double bachelor degree in Chemistry and Biology
Defense date
07.16.2025 15:30
07.16.2025 15:30
Summary
Antibiotic resistance is a current issue that effects all regions and socioeconomic levels. One of the new approaches being studied for the controlled administration of these drugs is the use os organic hydrogels, due to properties such as their biocompatibility, biodegradabilitiy, and ability to encapsulate various antibiotics. This Final Degree Project aims to study the bacterricidal capacity of a gel formed from a derivate of shikimic acid. The encapsulation of two different groups of antibiotics was carried outs: beta-lactams (ampicilin) and quinolones (nalidixic acid), and various variables were analysed to characterise the drug-release potential of the gel, as well as the bactericidal capacity of the gel itself. Controlled release of the encapsulated drug was observed through agar diffusion tests with two bacterial species under evaluation (Escherichia coli anda Staphylococcus epidermidis). another variable analysed was the potential effect of incubation time and temperatura on the release of the encapsulated antibiotic. It was concluded that gels formed by thermal shock and sonication do not exhibit intrinsic bactericidal activity, and the release capacity of the formed cogel was confirmed. An effect of incubation time and temperature on the inhibitory effect was detected, with a prior incubation at fifteen degrees Celsius for twelve hours allowing the cogel to display a greater bactericidal effect. Overall results showed a greater sensitivity of E. coli to the antibiotics.
Antibiotic resistance is a current issue that effects all regions and socioeconomic levels. One of the new approaches being studied for the controlled administration of these drugs is the use os organic hydrogels, due to properties such as their biocompatibility, biodegradabilitiy, and ability to encapsulate various antibiotics. This Final Degree Project aims to study the bacterricidal capacity of a gel formed from a derivate of shikimic acid. The encapsulation of two different groups of antibiotics was carried outs: beta-lactams (ampicilin) and quinolones (nalidixic acid), and various variables were analysed to characterise the drug-release potential of the gel, as well as the bactericidal capacity of the gel itself. Controlled release of the encapsulated drug was observed through agar diffusion tests with two bacterial species under evaluation (Escherichia coli anda Staphylococcus epidermidis). another variable analysed was the potential effect of incubation time and temperatura on the release of the encapsulated antibiotic. It was concluded that gels formed by thermal shock and sonication do not exhibit intrinsic bactericidal activity, and the release capacity of the formed cogel was confirmed. An effect of incubation time and temperature on the inhibitory effect was detected, with a prior incubation at fifteen degrees Celsius for twelve hours allowing the cogel to display a greater bactericidal effect. Overall results showed a greater sensitivity of E. coli to the antibiotics.
Direction
LEMOS RAMOS, MANUEL LUIS (Tutorships)
AFONSO LAGES, MARTA CAROLINA (Co-tutorships)
LEMOS RAMOS, MANUEL LUIS (Tutorships)
AFONSO LAGES, MARTA CAROLINA (Co-tutorships)
Court
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
ESTEVEZ TORANZO, MARIA ALICIA CAROLINA (Chairman)
COVELO ARTOS, GUILLERMO (Secretary)
CANDAL SUAREZ, EVA MARIA (Member)
Bolaamphiphilic alkylbisamide hydrogels derived from (-)-shikimic acid with even-length hydrocarbonated spacer: New nanostructured materials for drug delivery
Authorship
L.S.M.
Double bachelor degree in Chemistry and Biology
L.S.M.
Double bachelor degree in Chemistry and Biology
Defense date
07.15.2025 09:00
07.15.2025 09:00
Summary
The identification of gels opened up a new and promising field of research in chemistry, due to their high versatility. The wide structural diversity that they can present, together with their capacity to retain substances in their interior, as well as their flexibility, biodegradability and biocompatibility, turn these materials into candidates of great interest for the development of applications in fields as diverse as the food, cosmetic, pharmaceutical, biomedical and environmental industries. This Final Degree Project focuses on the study of low molecular weight organic gellants (LMWOGs). In particular, it is carried out the synthesis of a bolaamphiphilic molecule derived from the shikimic acid-(-)-shikimic acid with an intermediate alkyl chain of 12 carbon atoms in length. Its gellating capacity in different solvents was evaluated by calculating the minimum gelation concentration in cases where gel is stably formed. In addition, this molecule’s potential as a controlled drug release system is studied by the formation of cogels with two broad-spectrum antibiotics (ampicillin and nalidixic acid) incorporated in its three-dimensional network.
The identification of gels opened up a new and promising field of research in chemistry, due to their high versatility. The wide structural diversity that they can present, together with their capacity to retain substances in their interior, as well as their flexibility, biodegradability and biocompatibility, turn these materials into candidates of great interest for the development of applications in fields as diverse as the food, cosmetic, pharmaceutical, biomedical and environmental industries. This Final Degree Project focuses on the study of low molecular weight organic gellants (LMWOGs). In particular, it is carried out the synthesis of a bolaamphiphilic molecule derived from the shikimic acid-(-)-shikimic acid with an intermediate alkyl chain of 12 carbon atoms in length. Its gellating capacity in different solvents was evaluated by calculating the minimum gelation concentration in cases where gel is stably formed. In addition, this molecule’s potential as a controlled drug release system is studied by the formation of cogels with two broad-spectrum antibiotics (ampicillin and nalidixic acid) incorporated in its three-dimensional network.
Direction
Estévez Cabanas, Juan Carlos (Tutorships)
QUIÑOA CABANA, EMILIO (Co-tutorships)
Estévez Cabanas, Juan Carlos (Tutorships)
QUIÑOA CABANA, EMILIO (Co-tutorships)
Court
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
GONZALEZ BELLO, CONCEPCION (Chairman)
SECO CASTRO, JOSÉ MANUEL (Secretary)
VAZQUEZ LOPEZ, MIGUEL (Member)
Grapevine transformation for induction of somatic embryogenesis
Authorship
P.D.S.P.
Bachelor of Biology
P.D.S.P.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
Grapevine transformation is a promising avenue for genetic improvement, hampered by the low efficiency of organogenesis and embryogenesis. This study explored the possibility of activating these processes by expressing developmental regulators, such as WUSCHEL2 and LEAFY COTYLEDON2, as these have proven successful in other species. Expression of these genes was achieved by assembling GreenGate constructs and using a transformation protocol using Agrobacterium tumefaciens in both grapevine and tobacco. Furthermore, the promoter of the OLEOSIN4 gene was used as a marker of somatic embryogenesis due to its high level of expression in embryogenic calli. The results indicated that, under the conditions tested, expression of LEAFY COTYLEDON2 was not sufficient to activate somatic embryogenesis, and that WUSCHEL2 did not induce direct organogenesis in grapevine. These findings underscore the difficulty of regeneration in the 'Thompson Seedless' and 'Albariño' varieties and the need to adjust factors such as tissue type, development stage, and the strength of the promoters used. Even so, the study lays the groundwork for the design of more effective genetic transformation protocols in grapevines, essential for their biotechnological improvement.
Grapevine transformation is a promising avenue for genetic improvement, hampered by the low efficiency of organogenesis and embryogenesis. This study explored the possibility of activating these processes by expressing developmental regulators, such as WUSCHEL2 and LEAFY COTYLEDON2, as these have proven successful in other species. Expression of these genes was achieved by assembling GreenGate constructs and using a transformation protocol using Agrobacterium tumefaciens in both grapevine and tobacco. Furthermore, the promoter of the OLEOSIN4 gene was used as a marker of somatic embryogenesis due to its high level of expression in embryogenic calli. The results indicated that, under the conditions tested, expression of LEAFY COTYLEDON2 was not sufficient to activate somatic embryogenesis, and that WUSCHEL2 did not induce direct organogenesis in grapevine. These findings underscore the difficulty of regeneration in the 'Thompson Seedless' and 'Albariño' varieties and the need to adjust factors such as tissue type, development stage, and the strength of the promoters used. Even so, the study lays the groundwork for the design of more effective genetic transformation protocols in grapevines, essential for their biotechnological improvement.
Direction
SAMPEDRO JIMÉNEZ, JAVIER (Tutorships)
SAMPEDRO JIMÉNEZ, JAVIER (Tutorships)
Court
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
GONZALEZ GONZALEZ, MARIA VICTORIA (Chairman)
TABOADA RODRIGUEZ, TERESA MARIA (Secretary)
ROMERO BUJAN, MARIA INMACULADA (Member)
Cetaceans of the Galician Coasts: Population Health, Potential Threats, and Conservation Strategies
Authorship
A.S.C.
Bachelor of Biology
A.S.C.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
Cetaceans play a key role in marine ecosystems as bioindicators of environmental health and trophic regulators. The Galician coast, known for its high biodiversity, hosts a wide range of species currently threatened by various anthropogenic pressures. This work presents a review of the historical and scientific knowledge on cetacean populations in Galicia, identifying major threats and assessing current and potential conservation strategies. The most significant risk factors include bycatch, increased maritime traffic, pollution (acoustic and chemical), and climate change. Priority measures include improving the management of Marine Protected Areas, developing mitigation technologies, and enforcing specific regulatory frameworks. The study also emphasizes the importance of systematic monitoring programs, environmental education, and public engagement. It provides an updated synthesis that can inform the development of conservation policies tailored to the regional context and aligned with international commitments to marine biodiversity.
Cetaceans play a key role in marine ecosystems as bioindicators of environmental health and trophic regulators. The Galician coast, known for its high biodiversity, hosts a wide range of species currently threatened by various anthropogenic pressures. This work presents a review of the historical and scientific knowledge on cetacean populations in Galicia, identifying major threats and assessing current and potential conservation strategies. The most significant risk factors include bycatch, increased maritime traffic, pollution (acoustic and chemical), and climate change. Priority measures include improving the management of Marine Protected Areas, developing mitigation technologies, and enforcing specific regulatory frameworks. The study also emphasizes the importance of systematic monitoring programs, environmental education, and public engagement. It provides an updated synthesis that can inform the development of conservation policies tailored to the regional context and aligned with international commitments to marine biodiversity.
Direction
GOMEZ RODRIGUEZ, CAROLA (Tutorships)
BASELGA FRAGA, ANDRES (Co-tutorships)
GOMEZ RODRIGUEZ, CAROLA (Tutorships)
BASELGA FRAGA, ANDRES (Co-tutorships)
Court
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
RETUERTO FRANCO, JOSE CARLOS RUBÉN (Chairman)
LORENZO CARBALLA, MARÍA OLALLA (Secretary)
MONTERROSO MARTINEZ, MARIA DEL CARMEN (Member)
Impact of Somatic LINE-1 Integration on RNA Structure in Human Tumors
Authorship
L.S.P.
Biotechnology Degree (2nd Ed. )
L.S.P.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
Transposable elements (TEs) are DNA sequences capable of changing their location within the genome that harbors them, a process known as transposition. TEs are classified into two main types based on their transposition mechanism: DNA transposons and RNA transposons, or retrotransposons. In mammals, they typically make up around 50% of the genome. As a final consequence of transposition, the integration of these sequences can cause genetic mutations, thereby influencing both species evolution and the development of diseases such as cancer. In fact, recent studies have shown that retrotransposition is highly active in certain human tumors. At the transcriptional level, it has been observed that retrotransposon insertions can alter RNA splicing or promote alternative transcription start or termination sites. However, RNA-level mutational mechanisms have been poorly studied, mainly due to limitations in short-read sequencing technologies (the most commonly used to date). This study aims to evaluate the impact of retrotransposon integration, primarily LINE-1 (L1) elements, on transcript structure in human tumors. To achieve this, we performed direct RNA sequencing using long-read technology, generating the transcriptomes of three human tumors with high rates of retrotransposon mobilization. We then analyzed these data using bioinformatic approaches to identify potential alterations in RNA processing. Our analysis identified somatic LINE-1 insertions associated with aberrant RNA structures, including retrotransposon exonization and premature transcription termination. These findings highlight the potential of somatic retrotransposon integration to impact gene expression and function in cancer, paving the way for future studies that further investigate these mutational mechanisms and their possible contribution to cancer development.
Transposable elements (TEs) are DNA sequences capable of changing their location within the genome that harbors them, a process known as transposition. TEs are classified into two main types based on their transposition mechanism: DNA transposons and RNA transposons, or retrotransposons. In mammals, they typically make up around 50% of the genome. As a final consequence of transposition, the integration of these sequences can cause genetic mutations, thereby influencing both species evolution and the development of diseases such as cancer. In fact, recent studies have shown that retrotransposition is highly active in certain human tumors. At the transcriptional level, it has been observed that retrotransposon insertions can alter RNA splicing or promote alternative transcription start or termination sites. However, RNA-level mutational mechanisms have been poorly studied, mainly due to limitations in short-read sequencing technologies (the most commonly used to date). This study aims to evaluate the impact of retrotransposon integration, primarily LINE-1 (L1) elements, on transcript structure in human tumors. To achieve this, we performed direct RNA sequencing using long-read technology, generating the transcriptomes of three human tumors with high rates of retrotransposon mobilization. We then analyzed these data using bioinformatic approaches to identify potential alterations in RNA processing. Our analysis identified somatic LINE-1 insertions associated with aberrant RNA structures, including retrotransposon exonization and premature transcription termination. These findings highlight the potential of somatic retrotransposon integration to impact gene expression and function in cancer, paving the way for future studies that further investigate these mutational mechanisms and their possible contribution to cancer development.
Direction
CASTRO TUBIO, JOSE MANUEL (Tutorships)
RODRIGUEZ CASTRO, JORGE (Co-tutorships)
CASTRO TUBIO, JOSE MANUEL (Tutorships)
RODRIGUEZ CASTRO, JORGE (Co-tutorships)
Court
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
CID FERNANDEZ, MARIA MAGDALENA (Chairman)
LÓPEZ FABAL, ADOLFO (Secretary)
HOSPIDO QUINTANA, ALMUDENA (Member)
Identification of lncRNAs involved in the development of liver disease
Authorship
P.S.G.
Biotechnology Degree (2nd Ed. )
P.S.G.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
Hepatic fibrosis is a progressive scarring process that disrupts the architecture and function of the liver. Hepatic stellate cells (HSCs) and transforming growth factor-beta (TGF-ß) are key players in this progression. Based on the hypothesis that certain long non-coding RNAs may modulate this activation, this study aimed to analyze the transcriptional changes induced by TGF-ß through RNA-Seq, using libraries generated via rRNA depletion in the LX-2 hepatic stellate cell line. Quality control analysis revealed a technical issue in library preparation, which led to the development of a more stringent and context-specific filtering method. This was made possible by the availability of equivalent data generated from polyadenylated transcript selection libraries. Ultimately, 2,651 differentially expressed genes were identified, 521 of which correspond to long non-coding RNAs. Notable findings include the overexpression of the mitochondrial gene MT-ND6 and the regulatory gene GPRACR, which is associated with collagen synthesis. Additionally, downregulation of histone-coding genes involved in chromatin organization was observed. These results contribute to a deeper understanding of the mechanisms driving hepatic fibrosis and highlight new diagnostic and therapeutic targets.
Hepatic fibrosis is a progressive scarring process that disrupts the architecture and function of the liver. Hepatic stellate cells (HSCs) and transforming growth factor-beta (TGF-ß) are key players in this progression. Based on the hypothesis that certain long non-coding RNAs may modulate this activation, this study aimed to analyze the transcriptional changes induced by TGF-ß through RNA-Seq, using libraries generated via rRNA depletion in the LX-2 hepatic stellate cell line. Quality control analysis revealed a technical issue in library preparation, which led to the development of a more stringent and context-specific filtering method. This was made possible by the availability of equivalent data generated from polyadenylated transcript selection libraries. Ultimately, 2,651 differentially expressed genes were identified, 521 of which correspond to long non-coding RNAs. Notable findings include the overexpression of the mitochondrial gene MT-ND6 and the regulatory gene GPRACR, which is associated with collagen synthesis. Additionally, downregulation of histone-coding genes involved in chromatin organization was observed. These results contribute to a deeper understanding of the mechanisms driving hepatic fibrosis and highlight new diagnostic and therapeutic targets.
Direction
VARELA REY, MARTA MARIA (Tutorships)
ESQUINAS ROMAN, EVA MARIA (Co-tutorships)
VARELA REY, MARTA MARIA (Tutorships)
ESQUINAS ROMAN, EVA MARIA (Co-tutorships)
Court
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
EIBES GONZALEZ, GEMMA MARIA (Chairman)
CASTRO TUBIO, JOSE MANUEL (Secretary)
GARCIA ALONSO, ANGEL (Member)
Organ-on-a-chip technology and
Authorship
A.T.G.
Biotechnology Degree (2nd Ed. )
A.T.G.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 12:30
07.17.2025 12:30
Summary
Neurodegenerative diseases have a high incidence and prevalence in the population. Their growth is exponential, due to the increase in life expectancy, since aging is the main risk factor for developing this type of pathology. Therefore, research and development of new methodologies to try to understand the etiology and mechanisms of these diseases or the improvement of pharmacological strategies for patients is a challenge for the scientific community. One of the most relevant issues is the improvement of disease models both in vitro or in vivo models. In this context, about 15 years ago, organ-on-a-chip was born as an innovative methodological tool that offers new possibilities to address these diseases and to be considered as a complementary method to the existing ones. The aim of this work is to describe organ-on-a-chip technology in depth, to determine its applicability in the field of neuroscience and to contrast the information with a reference researcher in the field. For this purpose, a literature review was carried out relying on publications collected in PubMed and Google Scholar databases between the years 2015-2025 and an interview with Prof. Josep M. Canals Coll. The results obtained indicate that organ-on-a-chip are three-dimensional devices that combine microfluidics with a wide variety of cell types. They are versatile, dynamic, reproducible and standardizable models for the replication of uni/multifactorial neurodegenerative diseases. Currently, they can be considered a complementary tool in biotoxicity and pharmacological safety studies together with animal models, although they are undoubtedly a very promising future perspective, for the pharmaceutical industry they still present important limitations for this approach.
Neurodegenerative diseases have a high incidence and prevalence in the population. Their growth is exponential, due to the increase in life expectancy, since aging is the main risk factor for developing this type of pathology. Therefore, research and development of new methodologies to try to understand the etiology and mechanisms of these diseases or the improvement of pharmacological strategies for patients is a challenge for the scientific community. One of the most relevant issues is the improvement of disease models both in vitro or in vivo models. In this context, about 15 years ago, organ-on-a-chip was born as an innovative methodological tool that offers new possibilities to address these diseases and to be considered as a complementary method to the existing ones. The aim of this work is to describe organ-on-a-chip technology in depth, to determine its applicability in the field of neuroscience and to contrast the information with a reference researcher in the field. For this purpose, a literature review was carried out relying on publications collected in PubMed and Google Scholar databases between the years 2015-2025 and an interview with Prof. Josep M. Canals Coll. The results obtained indicate that organ-on-a-chip are three-dimensional devices that combine microfluidics with a wide variety of cell types. They are versatile, dynamic, reproducible and standardizable models for the replication of uni/multifactorial neurodegenerative diseases. Currently, they can be considered a complementary tool in biotoxicity and pharmacological safety studies together with animal models, although they are undoubtedly a very promising future perspective, for the pharmaceutical industry they still present important limitations for this approach.
Direction
DIAZ RUIZ, MARIA DEL CARMEN (Tutorships)
LOPEZ LOPEZ, ANDREA (Co-tutorships)
DIAZ RUIZ, MARIA DEL CARMEN (Tutorships)
LOPEZ LOPEZ, ANDREA (Co-tutorships)
Court
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
Expression of proteins associated with seipina in primary fibroblasts of patients with PELD
Authorship
G.V.A.
Biotechnology Degree (2nd Ed. )
G.V.A.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 12:30
07.17.2025 12:30
Summary
Lipodystrophies are a group of rare diseases characterized by the abnormal and progressive loss of adipose tissue. Generalized congenital lipodystrophy type 2 is caused by variants in the BSCL2 gene, which encodes the seipin protein, and is closely associated with Celia encephalopathy, a childhood neurodegenerative disease. The expression and accurate detection of seipin is technically complex. The main objective of this work is to develop an efficient and reliable method to evaluate the expression and detection of seipin in primary fibroblasts derived from patients with Celia encephalopathy, in order to further investigate its function and pathological role. For this purpose, primary fibroblast cultures from three patients and one control subject were established. Subsequently, protein extraction and quantification was performed, as well as the comparison of two Western blot transfer systems: the traditional method and the Bis-Tris system. In addition, various commercial antibodies were evaluated, along with two custom antibodies, to determine which one could reliably quantify seipin. The results indicated that the Bis-Tris system provided greater efficiency, and that only one of the antibodies (the LI International short antigen, specific for the mutated isoform) allowed the identification of the expected protein bands.
Lipodystrophies are a group of rare diseases characterized by the abnormal and progressive loss of adipose tissue. Generalized congenital lipodystrophy type 2 is caused by variants in the BSCL2 gene, which encodes the seipin protein, and is closely associated with Celia encephalopathy, a childhood neurodegenerative disease. The expression and accurate detection of seipin is technically complex. The main objective of this work is to develop an efficient and reliable method to evaluate the expression and detection of seipin in primary fibroblasts derived from patients with Celia encephalopathy, in order to further investigate its function and pathological role. For this purpose, primary fibroblast cultures from three patients and one control subject were established. Subsequently, protein extraction and quantification was performed, as well as the comparison of two Western blot transfer systems: the traditional method and the Bis-Tris system. In addition, various commercial antibodies were evaluated, along with two custom antibodies, to determine which one could reliably quantify seipin. The results indicated that the Bis-Tris system provided greater efficiency, and that only one of the antibodies (the LI International short antigen, specific for the mutated isoform) allowed the identification of the expected protein bands.
Direction
RODRIGUEZ REQUENA, JESUS (Tutorships)
ARAUJO VILAR, DAVID (Co-tutorships)
SANCHEZ IGLESIAS, SOFIA (Co-tutorships)
RODRIGUEZ REQUENA, JESUS (Tutorships)
ARAUJO VILAR, DAVID (Co-tutorships)
SANCHEZ IGLESIAS, SOFIA (Co-tutorships)
Court
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
Stress epigenetics: mechanisms and evidence of generational transmission
Authorship
J.V.C.
Bachelor of Biology
J.V.C.
Bachelor of Biology
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Environmental factors can induce epigenetic modifications that alter gene expression without changing the DNA sequence, with stress standing out due to its current impact on mental health. As long as these modifications escape the two reprogramming processes in the germline of each generation, they can affect both intergenerationally and transgenerationally, which could contribute to a predisposition to psychological disorders in future generations. In this way, the widespread acceptance that stress leaves an epigenetic mark affecting subsequent generations is explored, analyzing the available evidence, the molecular mechanisms involved (DNA methylation, histone modification, and non-coding RNA), and how they work, as well as the factors that influence their appearance and transmission. The literature review shows that exposure to environmental factors during windows of vulnerability, when the epigenome is especially sensitive, increases the likelihood that epigenetic modifications will affect the germline. However, while the reviewed studies support intergenerational epigenetic inheritance in both animal models and humans, only one experiment in rats provides direct molecular evidence of a transgenerational epigenetic transmission caused by stress.
Environmental factors can induce epigenetic modifications that alter gene expression without changing the DNA sequence, with stress standing out due to its current impact on mental health. As long as these modifications escape the two reprogramming processes in the germline of each generation, they can affect both intergenerationally and transgenerationally, which could contribute to a predisposition to psychological disorders in future generations. In this way, the widespread acceptance that stress leaves an epigenetic mark affecting subsequent generations is explored, analyzing the available evidence, the molecular mechanisms involved (DNA methylation, histone modification, and non-coding RNA), and how they work, as well as the factors that influence their appearance and transmission. The literature review shows that exposure to environmental factors during windows of vulnerability, when the epigenome is especially sensitive, increases the likelihood that epigenetic modifications will affect the germline. However, while the reviewed studies support intergenerational epigenetic inheritance in both animal models and humans, only one experiment in rats provides direct molecular evidence of a transgenerational epigenetic transmission caused by stress.
Direction
CANDAL SUAREZ, EVA MARIA (Tutorships)
CANDAL SUAREZ, EVA MARIA (Tutorships)
Court
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
In silico analysis of the expression of epitranscriptomic regulators in progeria (HGPS) and generation of a molecular tool for its in vitro study
Authorship
P.V.I.
Bachelor of Biology
P.V.I.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
Hutchinson-Guilford Progeria Syndrome, a rare disease characterized by premature aging, has emerged as a promising model for the study of physiological aging over the past few decades. At the same time, in recent years, technological advances have propelled the study of RNA chemical modifications in the transcriptome (epitranscriptomics), demonstrating their link to an increasing number of pathologies. Thus, in this study, differences in the global expression and the expression of epitranscriptomic regulators were analyzed between fibroblasts from patients with HGPS and young, healthy patients. To this end, a bioinformatic analysis was performed using previously published RNA mass sequencing data. Significant differential expression of the RNA modifiers ADARB1, THUMPD2, and TRMT9B was found, making them candidates for future aging and progeria studies. To facilitate further in vitro studies, the final part of this dissertation focused on the generation of a molecular tool for the transient expression of progerin: the aberrant isoform of lamin A that causes HGPS.
Hutchinson-Guilford Progeria Syndrome, a rare disease characterized by premature aging, has emerged as a promising model for the study of physiological aging over the past few decades. At the same time, in recent years, technological advances have propelled the study of RNA chemical modifications in the transcriptome (epitranscriptomics), demonstrating their link to an increasing number of pathologies. Thus, in this study, differences in the global expression and the expression of epitranscriptomic regulators were analyzed between fibroblasts from patients with HGPS and young, healthy patients. To this end, a bioinformatic analysis was performed using previously published RNA mass sequencing data. Significant differential expression of the RNA modifiers ADARB1, THUMPD2, and TRMT9B was found, making them candidates for future aging and progeria studies. To facilitate further in vitro studies, the final part of this dissertation focused on the generation of a molecular tool for the transient expression of progerin: the aberrant isoform of lamin A that causes HGPS.
Direction
FIDALGO PEREZ, MIGUEL ANGEL (Tutorships)
GUALLAR ARTAL, DIANA (Co-tutorships)
FIDALGO PEREZ, MIGUEL ANGEL (Tutorships)
GUALLAR ARTAL, DIANA (Co-tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Generation of a deletion mutant of the tssM2 gene from the type VI secretion system T6SS in Vibrio europaeus by targeted mutagenesis for functional analysis of the system
Authorship
J.V.L.
Biotechnology Degree (2nd Ed. )
J.V.L.
Biotechnology Degree (2nd Ed. )
Defense date
07.16.2025 10:30
07.16.2025 10:30
Summary
Bivalve hatcheries are currently one of the main aquaculture activities, essential to meet the growing demand for food. However, the development of this sector is threatened by the emergence of infectious outbreaks such as vibriosis, caused by bacteria of the genus Vibrio, which lead to high mortality rates in larval cultures, including the emerging pathogen Vibrio europaeus. It has been previously described that its genome contains two type VI secretion systems, T6SS1 and T6SS2, involved in interbacterial competition and cytotoxicity, respectively, although their functional role remains uncharacterized. This study proposes the generation of a genetic deletion mutant of the tssM2 gene, which encodes an essential structural protein for the function of T6SS2, using a two step allelic exchange technique based on double homologous recombination. For this purpose, a recombinant suicide plasmid named pSW7848T PC19 was constructed using Gibson assembly, carrying homologous flanks to the target gene. The construct was validated by PCR, digestion with BamH1 and Sanger sequencing, transformed and amplified in Escherichia coli pi 3813 and subsequently mobilized into Vibrio europaeus via conjugation with the donor strain Escherichia coli beta 3914. Selection of merodiploids was performed using chloramphenicol, first homologous recombination, and final resolution was achieved by arabinose induced plasmid excision, second homologous recombination, generating clean knock out deletion mutants. Deletion of the gene was confirmed by phenotypic analysis, PCR with external primers, and Sanger sequencing of the mutant genome. The delta tssM2 mutant obtained represents a key molecular tool to support future functional studies on the role of T6SS2 in virulence, pathogenicity, and interbacterial competition in Vibrio europaeus, contributing to the development of preventive strategies against vibriosis in aquaculture.
Bivalve hatcheries are currently one of the main aquaculture activities, essential to meet the growing demand for food. However, the development of this sector is threatened by the emergence of infectious outbreaks such as vibriosis, caused by bacteria of the genus Vibrio, which lead to high mortality rates in larval cultures, including the emerging pathogen Vibrio europaeus. It has been previously described that its genome contains two type VI secretion systems, T6SS1 and T6SS2, involved in interbacterial competition and cytotoxicity, respectively, although their functional role remains uncharacterized. This study proposes the generation of a genetic deletion mutant of the tssM2 gene, which encodes an essential structural protein for the function of T6SS2, using a two step allelic exchange technique based on double homologous recombination. For this purpose, a recombinant suicide plasmid named pSW7848T PC19 was constructed using Gibson assembly, carrying homologous flanks to the target gene. The construct was validated by PCR, digestion with BamH1 and Sanger sequencing, transformed and amplified in Escherichia coli pi 3813 and subsequently mobilized into Vibrio europaeus via conjugation with the donor strain Escherichia coli beta 3914. Selection of merodiploids was performed using chloramphenicol, first homologous recombination, and final resolution was achieved by arabinose induced plasmid excision, second homologous recombination, generating clean knock out deletion mutants. Deletion of the gene was confirmed by phenotypic analysis, PCR with external primers, and Sanger sequencing of the mutant genome. The delta tssM2 mutant obtained represents a key molecular tool to support future functional studies on the role of T6SS2 in virulence, pathogenicity, and interbacterial competition in Vibrio europaeus, contributing to the development of preventive strategies against vibriosis in aquaculture.
Direction
DUBERT PEREZ, JAVIER (Tutorships)
VENCES LORENZO, ANA (Co-tutorships)
DUBERT PEREZ, JAVIER (Tutorships)
VENCES LORENZO, ANA (Co-tutorships)
Court
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
López Romalde, Jesús Ángel (Chairman)
FRANCO RUIZ, DANIEL JOSE (Secretary)
GARCIA JARES, CARMEN MARIA (Member)
Study of enteric protozoa in the red-legged partridge (Alectoris rufa Linnaeus, 1758)
Authorship
L.V.M.
Bachelor of Biology
L.V.M.
Bachelor of Biology
Defense date
07.16.2025 10:00
07.16.2025 10:00
Summary
The presence of Eimeria, Cryptosporidium and Giardia in the intestinal contents of 22 specimens of red-legged partridge (Alectoris rufa Linnaeus, 1758) from two hunting grounds in Galicia was investigated. Samples were individually homogenized in a mortar, filtered, and concentrated by centrifugation in 0.04 M pH 7.2/diethyl ether (2:1) phosphate buffer saline. Aliquots of 10 microL of the sediments obtained were observed under brightfield microscopy. In parallel, 50 microL of the sediments were subjected to a direct immunofluorescence technique to detect Cryptosporidium and Giardia oo/cysts. In addition, nucleic acids were extracted from 200 microL of the sediments and a polymerase chain reaction (PCR) technique was applied to amplify fragments of the SSU-rRNA gene from Eimeria (420 bp), Cryptosporidium (587 bp) and Giardia (175 bp). Thus, in the 22 samples analyzed, Eimeria was detected in 14 (63.6%), identifying Eimeria chapmani, Eimeria hargisi, Eimeria kofoidi, Eimeria legionensis and a species of Eimeria similar to Eimeria maxima. Cryptosporidium parvum was found in three samples (13.6%) and subsequent analysis by PCR and sequencing of a fragment encoding GP60 gene allowed the identification of the zoonotic subtypes IIaA15G2R1 and IIaA16G3R1. All attempts to amplify Giardia in the samples were unsuccessful. The present study contributes to the knowledge about Eimeria and Cryptosporidium in wild galliform species, molecularly characterizing for the first time several species of Eimeria and C. parvum in A. rufa in Spain. On the other hand, the presence of C. parvum zoonotic subtypes in red-legged partridge demonstrates the wide distribution of this apicomplexan in wild animals and highlights the possible role of this bird in the transmission of cryptosporidiosis.
The presence of Eimeria, Cryptosporidium and Giardia in the intestinal contents of 22 specimens of red-legged partridge (Alectoris rufa Linnaeus, 1758) from two hunting grounds in Galicia was investigated. Samples were individually homogenized in a mortar, filtered, and concentrated by centrifugation in 0.04 M pH 7.2/diethyl ether (2:1) phosphate buffer saline. Aliquots of 10 microL of the sediments obtained were observed under brightfield microscopy. In parallel, 50 microL of the sediments were subjected to a direct immunofluorescence technique to detect Cryptosporidium and Giardia oo/cysts. In addition, nucleic acids were extracted from 200 microL of the sediments and a polymerase chain reaction (PCR) technique was applied to amplify fragments of the SSU-rRNA gene from Eimeria (420 bp), Cryptosporidium (587 bp) and Giardia (175 bp). Thus, in the 22 samples analyzed, Eimeria was detected in 14 (63.6%), identifying Eimeria chapmani, Eimeria hargisi, Eimeria kofoidi, Eimeria legionensis and a species of Eimeria similar to Eimeria maxima. Cryptosporidium parvum was found in three samples (13.6%) and subsequent analysis by PCR and sequencing of a fragment encoding GP60 gene allowed the identification of the zoonotic subtypes IIaA15G2R1 and IIaA16G3R1. All attempts to amplify Giardia in the samples were unsuccessful. The present study contributes to the knowledge about Eimeria and Cryptosporidium in wild galliform species, molecularly characterizing for the first time several species of Eimeria and C. parvum in A. rufa in Spain. On the other hand, the presence of C. parvum zoonotic subtypes in red-legged partridge demonstrates the wide distribution of this apicomplexan in wild animals and highlights the possible role of this bird in the transmission of cryptosporidiosis.
Direction
ARES MAZAS, MARIA ELVIRA (Tutorships)
COUSO PEREZ, SEILA (Co-tutorships)
ARES MAZAS, MARIA ELVIRA (Tutorships)
COUSO PEREZ, SEILA (Co-tutorships)
Court
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
LEMOS RAMOS, MANUEL LUIS (Chairman)
PORTEIRO COUTO, BEGOÑA (Secretary)
Rodriguez Diaz, Miguel Angel (Member)
Alterations in the blood-brain barrier (BBB) in cancer-associated cachexia
Authorship
L.V.V.
Biotechnology Degree (2nd Ed. )
L.V.V.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Cancer-associated cachexia is a multifactorial syndrome that alters the patient's metabolic state. The systemic inflammatory response induced by the tumour can affect the integrity of the blood-brain barrier (BBB), causing a dysregulation of the central nervous system that contributes to the progression of cachexia. Understanding the relationship between cancer-associated cachexia and the BBB could contribute to improving its clinical management and reducing its potential neurological complications. The hypothesis of this study is that a simple in vitro model of the BBB allows the study of alterations induced by circulating factors present in the serum of mice with cancer-induced cachexia. The aim of this study was to evaluate the effect of these sera on the structure of the BHE. To this end, an in vitro model of BHE based on bEnd.3 cells treated with sera from mice with Lewis lung carcinoma or control mouse sera was used. Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT), reactive oxygen species (ROS) production by flow cytometry, alterations in the endothelial barrier structure by determining transendothelial electrical resistance (TEER) and tight junction (TJ) protein expression by Western blot. The main results indicate that, at the concentration used, sera from mice with cachexia do not induce a significant decrease in cell viability, do not cause a significant increase in ROS production, but tend to reduce TEER after prolonged exposure of endothelial cells, without generating significant changes in tight junction protein expression. Therefore, the hypothesis could not be definitively validated, and studies with greater statistical power using different serum concentrations are needed to obtain more conclusive results.
Cancer-associated cachexia is a multifactorial syndrome that alters the patient's metabolic state. The systemic inflammatory response induced by the tumour can affect the integrity of the blood-brain barrier (BBB), causing a dysregulation of the central nervous system that contributes to the progression of cachexia. Understanding the relationship between cancer-associated cachexia and the BBB could contribute to improving its clinical management and reducing its potential neurological complications. The hypothesis of this study is that a simple in vitro model of the BBB allows the study of alterations induced by circulating factors present in the serum of mice with cancer-induced cachexia. The aim of this study was to evaluate the effect of these sera on the structure of the BHE. To this end, an in vitro model of BHE based on bEnd.3 cells treated with sera from mice with Lewis lung carcinoma or control mouse sera was used. Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT), reactive oxygen species (ROS) production by flow cytometry, alterations in the endothelial barrier structure by determining transendothelial electrical resistance (TEER) and tight junction (TJ) protein expression by Western blot. The main results indicate that, at the concentration used, sera from mice with cachexia do not induce a significant decrease in cell viability, do not cause a significant increase in ROS production, but tend to reduce TEER after prolonged exposure of endothelial cells, without generating significant changes in tight junction protein expression. Therefore, the hypothesis could not be definitively validated, and studies with greater statistical power using different serum concentrations are needed to obtain more conclusive results.
Direction
VIÑA CASTELAO, MARÍA DOLORES (Tutorships)
SEÑARIS RODRIGUEZ, ROSA MARIA (Co-tutorships)
VIÑA CASTELAO, MARÍA DOLORES (Tutorships)
SEÑARIS RODRIGUEZ, ROSA MARIA (Co-tutorships)
Court
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
MARTINEZ COSTAS, JOSE MANUEL (Chairman)
GARCIA FANDIÑO, REBECA (Secretary)
CSABA , NOEMI STEFANIA (Member)
Evaluation of the antiviral activity of different ionic liquids against the nervous necrosis virus (NNV) and the spring viremia of carp virus (SVCV)
Authorship
M.M.V.B.
Bachelor of Biology
M.M.V.B.
Bachelor of Biology
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Ionic liquids (ILs) are compounds composed exclusively of ions, with physicochemical properties that make them potentially useful in biomedical applications. In this study, the antiviral activity of two ionic liquids, Betaine TFSI and AMIM Cl, was evaluated against two viruses relevant to aquaculture: the nervous necrosis virus (NNV), a non-enveloped virus, and the spring viremia of carp virus (SVCV), an enveloped virus. Cytotoxicity assays were conducted on specific cell lines, along with tests to assess inhibition of viral adsorption and replication. The findings demonstrate that certain ionic liquids have the capacity to interfere with the viral replication cycle, suggesting their potential as innovative tools for controlling infectious diseases in aquaculture.
Ionic liquids (ILs) are compounds composed exclusively of ions, with physicochemical properties that make them potentially useful in biomedical applications. In this study, the antiviral activity of two ionic liquids, Betaine TFSI and AMIM Cl, was evaluated against two viruses relevant to aquaculture: the nervous necrosis virus (NNV), a non-enveloped virus, and the spring viremia of carp virus (SVCV), an enveloped virus. Cytotoxicity assays were conducted on specific cell lines, along with tests to assess inhibition of viral adsorption and replication. The findings demonstrate that certain ionic liquids have the capacity to interfere with the viral replication cycle, suggesting their potential as innovative tools for controlling infectious diseases in aquaculture.
Direction
BANDIN MATOS, MARIA ISABEL (Tutorships)
SOUTO PEREIRA, SANDRA (Co-tutorships)
BANDIN MATOS, MARIA ISABEL (Tutorships)
SOUTO PEREIRA, SANDRA (Co-tutorships)
Court
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
LAMAS FERNANDEZ, JESUS (Chairman)
DUBERT PEREZ, JAVIER (Secretary)
González Blanco, Miguel (Member)
Multimerization of HIV Glycoprotein Epitopes Eliciting Broadly Neutralizing Antibodies Using IC-Tagging
Authorship
N.V.A.
Biotechnology Degree (2nd Ed. )
N.V.A.
Biotechnology Degree (2nd Ed. )
Defense date
07.17.2025 12:30
07.17.2025 12:30
Summary
The human immunodeficiency virus (HIV) remains one of the main challenges in global public health due to its high genetic variability and the difficulty in developing an effective vaccine. One of the most promising approaches focuses on the induction of broadly neutralizing antibodies (BNAs), capable of recognizing conserved epitopes of the viral envelope glycoprotein. However, these epitopes have low immunogenicity, which limits their ability to elicit an effective immune response. In this work, a system based on IC-Tagging technology was adapted to promote epitope multimerization and enhance their presentation to the immune system. To this end, two constructs containing HIV epitopes recognized by BNAs were designed, expressed using the baculovirus expression system in insect cells, and encapsulated in spheres formed by the system. The production of recombinant baculoviruses was carried out using bacmids confirmed by PCR and the expression of the proteins was confirmed by SDS-PAGE, Western Blot, and immunofluorescence. The analysis of the results confirmed the correct expression of the target proteins and their efficient encapsulation in the spheres formed by muNS-Mi and sec-muNS-Mi. These findings validate the IC-Tagging system as an effective platform for epitope multimerization, laying the groundwork for future developments of more potent immunogens against the human immunodeficiency virus.
The human immunodeficiency virus (HIV) remains one of the main challenges in global public health due to its high genetic variability and the difficulty in developing an effective vaccine. One of the most promising approaches focuses on the induction of broadly neutralizing antibodies (BNAs), capable of recognizing conserved epitopes of the viral envelope glycoprotein. However, these epitopes have low immunogenicity, which limits their ability to elicit an effective immune response. In this work, a system based on IC-Tagging technology was adapted to promote epitope multimerization and enhance their presentation to the immune system. To this end, two constructs containing HIV epitopes recognized by BNAs were designed, expressed using the baculovirus expression system in insect cells, and encapsulated in spheres formed by the system. The production of recombinant baculoviruses was carried out using bacmids confirmed by PCR and the expression of the proteins was confirmed by SDS-PAGE, Western Blot, and immunofluorescence. The analysis of the results confirmed the correct expression of the target proteins and their efficient encapsulation in the spheres formed by muNS-Mi and sec-muNS-Mi. These findings validate the IC-Tagging system as an effective platform for epitope multimerization, laying the groundwork for future developments of more potent immunogens against the human immunodeficiency virus.
Direction
MARTINEZ COSTAS, JOSE MANUEL (Tutorships)
BARREIRO PIÑEIRO, NATALIA (Co-tutorships)
MARTINEZ COSTAS, JOSE MANUEL (Tutorships)
BARREIRO PIÑEIRO, NATALIA (Co-tutorships)
Court
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)
FIGUEIRAS GUZMAN, ADOLFO (Chairman)
Pozo Míguez, Iago (Secretary)
DEL PINO GONZALEZ DE LA HIGUERA, PABLO ALFONSO (Member)