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)
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)
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)
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)
Increasing temperature and its effects on the recombination of betanodavirus genomic segments
Authorship
M.C.G.
Bachelor of Biology
M.C.G.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
The nervous necrosis virus (NNV) is the causative agent of viral encephalopathy and retinopathy in numerous fish species, it is also denominate VER or VNN (viral nervous necrosis). Throughout its history, this virus has caused significant mortality rates and economic losses in aquaculture. It is a member of the family Nodaviridae and the genus Betanodavirus, a non-enveloped virus whose segmented genome consists of two positive-sense single-stranded RNA molecules. The larger segment, RNA1, encodes the viral polymerase (protein A) and a subgenomic RNA (RNA3) that synthesizes two non-structural proteins (B1 and B2), while RNA2 encodes the capsid proteins. The classification of betanodaviruses is based on a sequence within RNA2, which establishes four genotypes: red-spotted grouper, striped jack, barfin flounder, and tiger puffer nervous necrosis virus (RGNNV, SJNNV, BFNNV, and TPNNV, respectively). Additionally, in southern Europe, recombinant forms SJNNV/RGNNV and RGNNV/SJNNV have been identified. The distribution of these four genotypes is determined by their tolerance to water temperature, a parameter that can be affected by climate change. In this study, an analysis was performed to determine whether temperature significantly influences the recombination of the SJNNV and RGNNV genotypes. For this purpose, coinfection with both viruses was carried out at 25 and 30 degree Celsius, the progeny was cloned and analyzed using quantitative PCR techniques. The results showed that a temperature of 25 degree favors the production of RNA1 from the SJNNV genotype and could promote the formation of SJNNV/RGNNV recombinants. In contrast, a temperature of 30 degree favors the dominance of the RGNNV/RGNNV genotype.
The nervous necrosis virus (NNV) is the causative agent of viral encephalopathy and retinopathy in numerous fish species, it is also denominate VER or VNN (viral nervous necrosis). Throughout its history, this virus has caused significant mortality rates and economic losses in aquaculture. It is a member of the family Nodaviridae and the genus Betanodavirus, a non-enveloped virus whose segmented genome consists of two positive-sense single-stranded RNA molecules. The larger segment, RNA1, encodes the viral polymerase (protein A) and a subgenomic RNA (RNA3) that synthesizes two non-structural proteins (B1 and B2), while RNA2 encodes the capsid proteins. The classification of betanodaviruses is based on a sequence within RNA2, which establishes four genotypes: red-spotted grouper, striped jack, barfin flounder, and tiger puffer nervous necrosis virus (RGNNV, SJNNV, BFNNV, and TPNNV, respectively). Additionally, in southern Europe, recombinant forms SJNNV/RGNNV and RGNNV/SJNNV have been identified. The distribution of these four genotypes is determined by their tolerance to water temperature, a parameter that can be affected by climate change. In this study, an analysis was performed to determine whether temperature significantly influences the recombination of the SJNNV and RGNNV genotypes. For this purpose, coinfection with both viruses was carried out at 25 and 30 degree Celsius, the progeny was cloned and analyzed using quantitative PCR techniques. The results showed that a temperature of 25 degree favors the production of RNA1 from the SJNNV genotype and could promote the formation of SJNNV/RGNNV recombinants. In contrast, a temperature of 30 degree favors the dominance of the RGNNV/RGNNV genotype.
Direction
BANDIN MATOS, MARIA ISABEL (Tutorships)
SOUTO PEREIRA, SANDRA (Co-tutorships)
BANDIN MATOS, MARIA ISABEL (Tutorships)
SOUTO PEREIRA, SANDRA (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)
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)
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)
Role of lncRNAs and RNA binding proteins (RBPs) in liver disease
Authorship
H.F.H.
Bachelor of Biology
H.F.H.
Bachelor of Biology
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Chronic liver disease generates a fibrogenic response in the liver as a result of hepatic stellate cell activation, resulting in an accumulation of extracellular matrix in the liver as a mechanism for repairing damaged tissue. This process is highly regulated at multiple levels; one of the least studied regulatory mechanisms is that exerted by some lncRNAs in the activation of hepatic stellate cells. This final degree project explores the functional role of LncRNA-C1 in the development of liver fibrosis, with the aim of assessing its potential use in therapies for chronic liver disease. With this objective in mind, the levels of marker proteins of hepatic stellate cell activation and liver fibrosis, such as collagen and alpha-SMA, are analysed using histological techniques and Western blotting in two mouse models, bile duct ligation, which mimics human hepatic cholestasis, and a methionine-restricted, choline-deficient diet, which mimics metabolic associated fatty liver disease in mice in which the LncRNA-C1 gene has been deleted. Previously, using transient gene silencing models, the group had demonstrated the involvement of the LncRNA-C1 gene in the development of liver damage and fibrosis in the murine model of cholestasis. In this study, we observed increased activation of hepatic stellate cells in vivo after bile duct ligation in mice lacking the LncRNA-C1 gene, using histological techniques, when this damage extends over a period of more than 20 days. Possible improvements to the protocols, as well as new models capable of producing greater chronic damage, are proposed at the end of this study to study the impact of the absence of LncRNA-C1 on the development of chronic liver disease.
Chronic liver disease generates a fibrogenic response in the liver as a result of hepatic stellate cell activation, resulting in an accumulation of extracellular matrix in the liver as a mechanism for repairing damaged tissue. This process is highly regulated at multiple levels; one of the least studied regulatory mechanisms is that exerted by some lncRNAs in the activation of hepatic stellate cells. This final degree project explores the functional role of LncRNA-C1 in the development of liver fibrosis, with the aim of assessing its potential use in therapies for chronic liver disease. With this objective in mind, the levels of marker proteins of hepatic stellate cell activation and liver fibrosis, such as collagen and alpha-SMA, are analysed using histological techniques and Western blotting in two mouse models, bile duct ligation, which mimics human hepatic cholestasis, and a methionine-restricted, choline-deficient diet, which mimics metabolic associated fatty liver disease in mice in which the LncRNA-C1 gene has been deleted. Previously, using transient gene silencing models, the group had demonstrated the involvement of the LncRNA-C1 gene in the development of liver damage and fibrosis in the murine model of cholestasis. In this study, we observed increased activation of hepatic stellate cells in vivo after bile duct ligation in mice lacking the LncRNA-C1 gene, using histological techniques, when this damage extends over a period of more than 20 days. Possible improvements to the protocols, as well as new models capable of producing greater chronic damage, are proposed at the end of this study to study the impact of the absence of LncRNA-C1 on the development of chronic liver disease.
Direction
VARELA REY, MARTA MARIA (Tutorships)
VARELA REY, MARTA 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)
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)
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)
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)
Optimization of a Method for Purification and Sequencing of Long DNA Fragments from Low-Cellularity Biological Samples
Authorship
A.G.L.
Biotechnology Degree (2nd Ed. )
A.G.L.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 10:00
07.18.2025 10:00
Summary
Long-read sequencing offers advantages for the analysis of structural variants and repetitive regions of the human genome, but its application to samples with low DNA content requires optimized protocols. This study hypothesizes that an appropriate combination of preservation, lysis and amplification methods would yield DNA of sufficient quality for third-generation sequencing. To test this, three tissue-preservation methods (freezing at -80 degrees Celsius, PAXgene fixation without paraffin and paraffin embedding after PAXgene treatment) were compared, two lysis strategies (proteinase K versus the commercial REPLI-g Single Cell kit) were evaluated, and a whole-genome amplification and library-preparation workflow designed to preserve DNA structure was applied. The methodology combined fluorometric quantification, integrity assessment via TapeStation electropherograms, isothermal amplification, enzymatic debranching, magnetic purification and library preparation for subsequent sequencing. The combination of freezing or paraffin-free fixation with proteinase K lysis preserved high-molecular-weight DNA fragments and optimal integrity. The amplification and cleanup strategy efficiently retained enriched DNA, and the MinION run generated data with adequate coverage and quality to identify structural variants. These findings validate the proposed protocol for low-input samples and lay the groundwork for its application in clinical settings and in studies based on long-read technologies.
Long-read sequencing offers advantages for the analysis of structural variants and repetitive regions of the human genome, but its application to samples with low DNA content requires optimized protocols. This study hypothesizes that an appropriate combination of preservation, lysis and amplification methods would yield DNA of sufficient quality for third-generation sequencing. To test this, three tissue-preservation methods (freezing at -80 degrees Celsius, PAXgene fixation without paraffin and paraffin embedding after PAXgene treatment) were compared, two lysis strategies (proteinase K versus the commercial REPLI-g Single Cell kit) were evaluated, and a whole-genome amplification and library-preparation workflow designed to preserve DNA structure was applied. The methodology combined fluorometric quantification, integrity assessment via TapeStation electropherograms, isothermal amplification, enzymatic debranching, magnetic purification and library preparation for subsequent sequencing. The combination of freezing or paraffin-free fixation with proteinase K lysis preserved high-molecular-weight DNA fragments and optimal integrity. The amplification and cleanup strategy efficiently retained enriched DNA, and the MinION run generated data with adequate coverage and quality to identify structural variants. These findings validate the proposed protocol for low-input samples and lay the groundwork for its application in clinical settings and in studies based on long-read technologies.
Direction
CASTRO TUBIO, JOSE MANUEL (Tutorships)
RODRIGUEZ CASTRO, JORGE (Co-tutorships)
CASTRO TUBIO, JOSE MANUEL (Tutorships)
RODRIGUEZ CASTRO, JORGE (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)
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)
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)
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)
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)
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 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)
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)
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)
Could the statistics of wolf damage to livestock be considered an adequate indicator of the impact on the livestock population?
Authorship
J.R.V.
Bachelor of Biology
J.R.V.
Bachelor of Biology
Defense date
07.17.2025 10:00
07.17.2025 10:00
Summary
There is data about the impact of wolves on the livestock, managed by the administration and generated from complaints and claims filed by livestock farmers, who are compensated for the damage caused. This study seeks to determine whether statistics on livestock damage caused by wolves can be considered an accurate indicator and whether more reliable alternative methods exist. The main objective is to evaluate these statistics and compare them with other methods, such as scat analysis to study wolf diet and the use of GPS collars. To this end, a literature review of studies on wolf predation and diet was conducted. Although damage statistics are useful, they do not always accurately reflect the true impact of wolves on livestock, as they can be influenced by factors such as fraudulent such as fraudulent complaints from ranchers or missing cattle, among others. Diet studies show that it is not possible to differentiate whether the cause of death of the prey was predation or scavenging, which makes it difficult to provide accurate data. However, monitoring wolves with GPS collars could produce much more reliable data in this regard, free from bias. This study would be useful to verify whether the money spent by the government on compensation for damages corresponds to the actual data of wolf predation on livestock.
There is data about the impact of wolves on the livestock, managed by the administration and generated from complaints and claims filed by livestock farmers, who are compensated for the damage caused. This study seeks to determine whether statistics on livestock damage caused by wolves can be considered an accurate indicator and whether more reliable alternative methods exist. The main objective is to evaluate these statistics and compare them with other methods, such as scat analysis to study wolf diet and the use of GPS collars. To this end, a literature review of studies on wolf predation and diet was conducted. Although damage statistics are useful, they do not always accurately reflect the true impact of wolves on livestock, as they can be influenced by factors such as fraudulent such as fraudulent complaints from ranchers or missing cattle, among others. Diet studies show that it is not possible to differentiate whether the cause of death of the prey was predation or scavenging, which makes it difficult to provide accurate data. However, monitoring wolves with GPS collars could produce much more reliable data in this regard, free from bias. This study would be useful to verify whether the money spent by the government on compensation for damages corresponds to the actual data of wolf predation on livestock.
Direction
DOMINGUEZ CONDE, JESUS (Tutorships)
LLANEZA RODRIGUEZ, LUIS ALADINO (Co-tutorships)
DOMINGUEZ CONDE, JESUS (Tutorships)
LLANEZA RODRIGUEZ, LUIS ALADINO (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)
Biorefinery design by modeling. Case study of brewer’s spent grain
Authorship
P.S.M.
Biotechnology Degree (2nd Ed. )
P.S.M.
Biotechnology Degree (2nd Ed. )
Defense date
07.18.2025 09:00
07.18.2025 09:00
Summary
This Bachelor's Thesis presents an approach to the preliminary design of a biorefinery, based on the definition and optimization of a superstructure. The case study focuses on brewer’s spent grain (BSG), the main by-product of the brewing industry. Through a bibliographic review, multiple valorization pathways were identified. The selected technologies, which were chosen in order to target the utilization of different fractions of BSG, were then characterized. These valorization alternatives were integrated into a superstructure, allowing the comparison of the various proposed processes for obtaining commercially valuable products. In this context, the use of BSG as a feedstock in microbially mediated transformations was assessed by applying genome-scale metabolic models (GEMs). These models enabled a screening of microorganisms reported in the literature, helping to identify potential carbon sources, possible products, and their respective yields. Finally, the superstructure optimization problem was formulated and solved using OUTDOOR, determining the most economically favorable pathway. Among the various solutions explored and compared, the most promising processes identified were: (1) direct composting of BSG, (2) extraction of phenolic compounds followed by anaerobic digestion of the remaining fraction, and (3) protein extraction combined with a biological transformation mediated by Saccharomyces cerevisiae. This methodology proved to be a robust, flexible, and replicable tool with strong potential for application in the early stages of biorefinery design aimed at the valorization of various agro-industrial residues.
This Bachelor's Thesis presents an approach to the preliminary design of a biorefinery, based on the definition and optimization of a superstructure. The case study focuses on brewer’s spent grain (BSG), the main by-product of the brewing industry. Through a bibliographic review, multiple valorization pathways were identified. The selected technologies, which were chosen in order to target the utilization of different fractions of BSG, were then characterized. These valorization alternatives were integrated into a superstructure, allowing the comparison of the various proposed processes for obtaining commercially valuable products. In this context, the use of BSG as a feedstock in microbially mediated transformations was assessed by applying genome-scale metabolic models (GEMs). These models enabled a screening of microorganisms reported in the literature, helping to identify potential carbon sources, possible products, and their respective yields. Finally, the superstructure optimization problem was formulated and solved using OUTDOOR, determining the most economically favorable pathway. Among the various solutions explored and compared, the most promising processes identified were: (1) direct composting of BSG, (2) extraction of phenolic compounds followed by anaerobic digestion of the remaining fraction, and (3) protein extraction combined with a biological transformation mediated by Saccharomyces cerevisiae. This methodology proved to be a robust, flexible, and replicable tool with strong potential for application in the early stages of biorefinery design aimed at the valorization of various agro-industrial residues.
Direction
MAURICIO IGLESIAS, MIGUEL (Tutorships)
MAURICIO IGLESIAS, 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)
Analysis of lymphocyte populations in the spleen and peritoneal cavity of vaccinated turbot
Authorship
L.S.M.
Bachelor of Biology
L.S.M.
Bachelor of Biology
Defense date
07.17.2025 09:30
07.17.2025 09:30
Summary
In this research work, with the purpose of analyzing the immune response in turbot after immunization with antigens of Philasterides dicentrarchi or ficoerythrin, a study of the populations of IgM+ B lymphocytes and T lymphocytes in spleen and vaccine-cell clusters of the peritoneal cavity was carried out on vaccinated turbot. To determine the presence of these lymphoid cells, immunofluorescence and in situ hybridization techniques were used to identify B IgM+ and T cells at 4, 7, 14 and 21 days after immunization. Immunization generated an intense proliferation of IgM+ B lymphocytes in the lymphoid tissue associated with the melanomacrophagic centers (M-LAs) from day 4, increasing until day 7. From day 14, the number and proliferative activity of B cells in M-LAs was reduced. As for the T cells, they take a peripheral position around the M-LAs, interacting with the outer layers of the B lymphocytes. On the other hand, the migration of B and T cells to the peritoneal cavity was observed, being located in thevaccine formed in it as a result of immunization.
In this research work, with the purpose of analyzing the immune response in turbot after immunization with antigens of Philasterides dicentrarchi or ficoerythrin, a study of the populations of IgM+ B lymphocytes and T lymphocytes in spleen and vaccine-cell clusters of the peritoneal cavity was carried out on vaccinated turbot. To determine the presence of these lymphoid cells, immunofluorescence and in situ hybridization techniques were used to identify B IgM+ and T cells at 4, 7, 14 and 21 days after immunization. Immunization generated an intense proliferation of IgM+ B lymphocytes in the lymphoid tissue associated with the melanomacrophagic centers (M-LAs) from day 4, increasing until day 7. From day 14, the number and proliferative activity of B cells in M-LAs was reduced. As for the T cells, they take a peripheral position around the M-LAs, interacting with the outer layers of the B lymphocytes. On the other hand, the migration of B and T cells to the peritoneal cavity was observed, being located in thevaccine formed in it as a result of immunization.
Direction
LAMAS FERNANDEZ, JESUS (Tutorships)
LAMAS FERNANDEZ, JESUS (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)
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)
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)