https://hdl.handle.net/10481/32992 2026-04-20T07:39:40Z 2026-04-20T07:39:40Z Even-Ros, Dilamm Huertas-Romero, Judit Marín-Menguiano, Miriam Nusspaumer, Gretel Borge, Miguel Irimia, Manuel Zurita Martínez, Federico González Reyes, Acaimo https://hdl.handle.net/10481/103976 2025-05-07T09:08:12Z

Drosophila ovarian stem cell niche ageing involves coordinated changes in transcription and alternative splicing Even-Ros, Dilamm; Huertas-Romero, Judit; Marín-Menguiano, Miriam; Nusspaumer, Gretel; Borge, Miguel; Irimia, Manuel; Zurita Martínez, Federico; González Reyes, Acaimo Gene expression (GE) and alternative splicing (AS) contribute to the formation of new interaction networks with potentially significant cellular functions. Here, we investigate ageing in the Drosophila female germline stem cell (GSC) niche and describe functional changes in both GE and AS. The GSC niche comprises three types of support cells, whose ageing transcriptomes reveal differential GE and AS variations related to cell adhesion, cytoskeleton and neural signalling. Because each population show distinctive GE and AS changes, niche cell types possess unique ageing signatures. Depending on the cell population, groups of genes display changes in both GE and AS, revealing a coordinated regulation of transcription and splicing during niche ageing. One such gene is Fasciclin 2, a neural adhesion molecule that we find is essential for niche functioning. Furthermore, genes involved in AS undergo changes in GE and/or AS themselves, providing a mechanistic explanation for the coordination of these two processes during niche ageing. This is the case of the splicing factor Smu1, described here as a key element necessary for ovarian niche homeostasis. The following grants provided financial support: PID2020-115040GB-I00 (MI), PID2021-125480NB-I00 and CEX2020-001088-M (A.G-.R.), funded by MICIU/AEI/10.13039/501100011033; P20_00888 funded by the Junta de Andalucía (A.G-.R.); and by ERDF, EU. D.E-R. (FPU15/06664) and J.H-R. (FPU17/03230) were funded by FPU contracts from the MECD.

Sánchez-Martín, Victoria Tello-López, María J. Ortiz-Morales, Andrea Murciano Calles, Javier García-Salcedo, José Antonio https://hdl.handle.net/10481/102981 2025-03-11T10:16:14Z

The histone deacetylase inhibitor Scriptaid targets G-quadruplexes Sánchez-Martín, Victoria; Tello-López, María J.; Ortiz-Morales, Andrea; Murciano Calles, Javier; García-Salcedo, José Antonio Scriptaid is a chemical compound with anti-tumoural effects due to its role as a histone deacetylase inhibitor. Despite sharing part of the chemical structure with other ligands of G-quadruplexes (G4s), the interaction of Scriptaid with G4s has not been explored before. We synthesized Scriptaid and screened its cytotoxic activity in cellular models of colorectal cancer (CRC). We extensively evaluated its biological activity by cell cycle, immunofluorescence, qRT-PCR and Western blot experiments. To identify the G4 targets of Scriptaid, we conducted a panel of binding assays. Here, we show that Scriptaid induced cytotoxicity, cell cycle arrest and nucleolar stress in CRC cells. Moreover, Scriptaid impaired RNA polymerase I (Pol I) transcription, stabilized G4s and caused DNA damage. Finally, we disclose that these effects were attributable to the binding of Scriptaid to G4s in ribosomal DNA. In conclusion, our work reveals that a primary impact of Scriptaid on human cells is the interaction with G4s. This study was funded by Ministry of Science and Innovation of Spain to J.A.G-S. (Instituto de Salud Carlos III (ISCIII), PI21/00497, co-funded by the European Union) and to M.S. (PID2020-120481RB-I00/AEI/10.13039/501100011033), by the local government Junta de Andalucía through PAIDI research groups (BIO-344) and by the Ministry of Science, Technological Development and Innovation of the Republic of Serbia (Grant Agreement with University of Belgrade-Faculty of Pharmacy No: 451‐03‐65/2024‐03/200161) to D.R. and K.N. The Government of Spain granted with PhD fellowship FPU16/05822 to V.S.-M. The University of Almeria granted with PhD fellowship CPRE2023-069 to M.J.T.-L.; Electronic supplementary material is available online at https://doi.org/10.6084/m9.figshare.c.7644194

Teso Pérez, Claudia López‑Gazcón, Areli Peralta Sánchez, Juan Manuel Martínez Bueno, Manuel Valdivia Martínez, Dolores Eva Farez Vidal, María Esther Martín Platero, Antonio Manuel https://hdl.handle.net/10481/102837 2025-03-04T10:28:56Z

Bacteriocin‑Producing Enterococci Modulate Cheese Microbial Diversity Teso Pérez, Claudia; López‑Gazcón, Areli; Peralta Sánchez, Juan Manuel; Martínez Bueno, Manuel; Valdivia Martínez, Dolores Eva; Farez Vidal, María Esther; Martín Platero, Antonio Manuel Cheese production involves various lactic acid bacteria (LAB) that break down lactose, milk proteins, and fats, producing key nutrients and influencing the cheese’s flavor. They form communities that play a crucial role in determining the cheese’s organoleptic properties. The composition of cheeses’ microbial communities is shaped by physicochemical factors (e.g., tem- perature, pH, and salinity) and biological factors (i.e. microbial interactions). While starter cultures are introduced to control these communities, non-starter LAB represent a significant portion of the final microbial assemblage, but their interactions remain unclear. LAB often produce bacteriocins, antimicrobial peptides that antagonize other bacteria, but their role within LAB communities is not fully understood. This study aimed to assess the impact of bacteriocin production on LAB diversity in cheese, using Enterococcus as a model organism, a common bacteriocin producer. We analyzed enterocin production of enterococcal isolates by antimicrobial assays and microbial diversity differences in raw milk cheeses by two approaches: 16S RNA gene amplicon metagenomic sequencing for the whole microbial community and multi-locus sequence analysis (MLSA) for the enterococcal diversity. Our results revealed that LAB communities were dominated by lactococci, lactobacilli, and streptococci, with enterococci present in lower numbers. However, cheeses containing bacteriocin-producing enterococci exhibited higher microbial diversity. Interestingly, the highest diversity occurred at low levels of bacteriocin producers, but this effect was not observed within enterococcal populations. These findings suggest that bacteriocin production plays a key role in shaping LAB communities during cheese ripening, although further research is needed to understand its broader implications in other microbial ecosystems. Funding for open access publishing: Universidad de Granada/CBUA. This research was funded by the Junta de Andalucía (Programa Operativo FEDER Andalucía 2014–2020), grant number A-BIO-083-UGR18 and PAIDI Program Group BIO 309 (MEF-V). C.T.-P. was funded by the Plan Estatal de Garantía Juvenil (Fondo Social Europeo, Gobierno de España, Ref. PEJ2018-003019-A.

Lozano Rodríguez, Noelia Gomez-Samblas, Mercedes Osuna Carrillo De Albornoz, Antonio https://hdl.handle.net/10481/86162 2023-12-13T13:22:23Z

Use of sera cell free DNA (cfDNA) and exovesicle-DNA for the molecular diagnosis of chronic Chagas disease Lozano Rodríguez, Noelia; Gomez-Samblas, Mercedes; Osuna Carrillo De Albornoz, Antonio Chagas disease, a neglected tropical disease, is now considered a worldwide health concern as a result of migratory movements from Central and South America to other regions that were considered free of the disease, and where the epidemiological risk is limited to transplacental transmission or blood or organ donations from infected persons. Parasite detection in chronically ill patients is restricted to serological tests that only determine infection by previous infection and not the presence of the parasite, especially in patients undergoing treatment evaluation or in newborns. We have evaluated the use of nucleic acids from both circulating exovesicles and cell-free DNA (cfDNA) from 50 samples twice randomly selected from a total of 448 serum samples from immunologically diagnosed patients in whom the presence of the parasite was confirmed by nested PCR on amplicons resulting from amplification with kinetoplastid DNA-specific primers 121F-122R. Six samples were randomly selected to quantify the limit of detection by qPCR in serum exovesicles. When the nucleic acids thus purified were assayed as a template and amplified with kinetoplastid DNA and nuclear satellite DNA primers, a 100% positivity rate was obtained for all positive samples assayed with kDNA-specific primers and 96% when SAT primers were used. However, isolation of cfDNA for Trypanosoma cruzi and amplification with SAT also showed 100% positivity. The results demonstrate that serum exovesicles contain DNA of mitochondrial and nuclear origin, which can be considered a mixed population of exovesicles of parasitic origin. The results obtained with serum samples prove that both cfDNA and Exovesicle DNA can be used to confirm parasitaemia in chronically ill patients or in samples where it is necessary to demonstrate the active presence of the parasite. The results confirm for the first time the existence of exovesicles of mitochondrial origin of the parasite in the serum of those affected by Chagas disease. This research was funded by the ERANet program, Research in prevention of congenital Chagas disease: parasitological, placental and immunological markers (ERANet17/HLH-0142 (Cochaco). Instituto Carlos III, Ministerio de Sanidad, Gobierno de Espana. Fundacion Ramon Areces "Interactoma de las exovesiculas de T. cruzi y de los inmunocomplejos que forman con las celulas del hospedador: implicaciones en la patologia de la enfermedad de Chagas (2019)". PreChag y el titulo Exovesiculas circulantes como marcadoras de diagnostico, PREcoz de la Enfermedad de CHAGas del XXI Concurso Nacional para la adjudicacion de Ayudas a la Investigacion en Ciencias de la Vida y de la Materia (2022). Ministerio de Ciencia y Tecnologia of the government of Spain funded the project PGC2018-099424-B-I00 and The financial support given by the proyect A-BIO-350-UGR18 I+D+i Proyect "Programa Operativo FEDER de Andalucia JJAA" 2014-2020.

Fernández Martínez, José Ramírez Casas, Yolanda Aranda Martínez, Paula López Rodríguez, Alba Escames Rosa, Germaine Acuña Castroviejo, Darío https://hdl.handle.net/10481/84956 2023-10-11T12:41:25Z

iMS-Bmal1−/− mice show evident signs of sarcopenia that are counteracted by exercise and melatonin therapies Fernández Martínez, José; Ramírez Casas, Yolanda; Aranda Martínez, Paula; López Rodríguez, Alba; Escames Rosa, Germaine; Acuña Castroviejo, Darío Sarcopenia is an age-related disease characterized by a reduction in muscle mass, strength, and function and, therefore, a deterioration in skeletal muscle health and frailty. Although the cause of sarcopenia is still unknown and, thus, there is no treatment, increasing evidence suggests that chronodisruption, particularly alterations in Bmal1 clock gene, can lead to those deficits culminating in sarcopenia. To gain insight into the cause and mechanism of sarcopenia and the protective effect of a therapeutic intervention with exercise and/or melatonin, the gastrocnemius muscles of male and female skeletal muscle-specific and inducible Bmal1 knockout mice (iMS-Bmal1−/−) were examined by phenotypic tests and light and electron microscopy. Our results revealed a disruption of the normal activity/rest rhythm, a drop in skeletal muscle function and mass, and increased frailty in male and female iMS-Bmal1−/− animals compared to controls. A reduction in muscle fiber size and increased collagenous tissue were also detected, accompanied by reduced mitochondrial oxidative capacity and a compensatory shift towards a more oxidative fiber type. Electron microscopy further supports mitochondrial impairment in mutant mice. Melatonin and exercise ameliorated the damage caused by loss of Bmal1 in mutant mice, except for mitochondrial damage, which was worsened by the latter. Thus, iMS-Bmal1−/− mice let us to identify Bmal1 deficiency as the responsible for the appearance of sarcopenia in the gastrocnemius muscle. Moreover, the results support the exercise and melatonin as therapeutic tools to counteract sarcopenia, by a mechanism that does not require the presence of Bmal1. This study was partially supported by grants from the Instituto de Salud Carlos III through the grants PI19‐01372 and CB/10/00238 (co‐funded by European Regional Development Fund/European Social Fund “Investing in your future”); the Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (CTS‐101), Spain. José Fernández‐Martínez is supported by an FPU fellowship from the Ministerio de Educación, Spain; Yolanda Ramírez‐Casas has a PFIS fellowship from the Instituto de Salud Carlos III; Paula Aranda‐Martínez has a fellowship from grant no. P18‐RT‐698, and Alba López‐Rodríguez has a fellowship from grant no. P18‐RT‐3222, from the Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía.