Instituto Universitario de Investigación de Biotecnologiahttps://hdl.handle.net/10481/329922024-03-28T13:52:47Z2024-03-28T13:52:47ZUse of sera cell free DNA (cfDNA) and exovesicle-DNA for the molecular diagnosis of chronic Chagas diseaseLozano Rodríguez, NoeliaGomez-Samblas, MercedesOsuna Carrillo De Albornoz, Antoniohttps://hdl.handle.net/10481/861622023-12-13T13:22:23ZUse 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.
iMS-Bmal1−/− mice show evident signs of sarcopenia that are counteracted by exercise and melatonin therapiesFernández Martínez, JoséRamírez Casas, YolandaAranda Martínez, PaulaLópez Rodríguez, AlbaEscames Rosa, GermaineAcuña Castroviejo, Daríohttps://hdl.handle.net/10481/849562023-10-11T12:41:25ZiMS-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.
Mucosal Immune Defence Gene Polymorphisms as Relevant Players in the Pathogenesis of IgA Vasculitis?Batista-Liz, Joao CarlosCarmona López, Francisco Davidhttps://hdl.handle.net/10481/849292023-11-28T23:21:04ZMucosal Immune Defence Gene Polymorphisms as Relevant Players in the Pathogenesis of IgA Vasculitis?
Batista-Liz, Joao Carlos; Carmona López, Francisco David
ITGAM–ITGAX (rs11150612, rs11574637), VAV3 rs17019602, CARD9 rs4077515, DEFA (rs2738048, rs10086568), and HORMAD2 rs2412971 are mucosal immune defence polymorphisms, that have an impact on IgA production, described as risk loci for IgA nephropathy (IgAN). Since IgAN and Immunoglobulin-A vasculitis (IgAV) share molecular mechanisms, with the aberrant deposit of IgA1 being the main pathophysiologic feature of both entities, we assessed the potential influence of the seven abovementioned polymorphisms on IgAV pathogenesis. These seven variants were genotyped in 381 Caucasian IgAV patients and 997 matched healthy controls. No statistically significant differences were observed in the genotype and allele frequencies of these seven polymorphisms when the whole cohort of IgAV patients and those with nephritis were compared to controls. Similar genotype and allele frequencies of all polymorphisms were disclosed when IgAV patients were stratified according to the age at disease onset or the presence/absence of gastrointestinal or renal manifestations. Likewise, no ITGAM–ITGAX and DEFA haplotype differences were observed when the whole cohort of IgAV patients, along with those with nephritis and controls, as well as IgAV patients, stratified according to the abovementioned clinical characteristics, were compared. Our results suggest that mucosal immune defence polymorphisms do not represent novel genetic risk factors for IgAV pathogenesis.
This research was funded by European Union FEDER funds and “Fondo de Investigaciones Sanitarias” from “Instituto de Salud Carlos III” (ISCIII, Health Ministry, Spain), grant numbers PI18/00042 and PI21/00042. J.C.B.-L. is a recipient of a PFIS program fellowship from the ISCIII, co-funded by the European Social Fund (‘Investing in your future’), grant number FI22/00020. M.S.M.-G. is supported by funds of “Fondo de Investigaciones Sanitarias” from ISCIII, grant number PI121/00042. R.L.-M. is a recipient of a Miguel Servet type II program fellowship from the ISCIII, co-funded by ESF (“Investing in your future”), grant number CPII21/00004.
Removal of the Water Pollutant Ciprofloxacin Using Biodegradable Sorbent Polymers Obtained from PolysaccharidesAlvarado, SarahMegía Fernández, AliciaOrtega Muñoz, MarianoHernández Mateo, FernandoLópez Jaramillo, Francisco JavierSantoyo González, Franciscohttps://hdl.handle.net/10481/846352023-09-25T10:19:03ZRemoval of the Water Pollutant Ciprofloxacin Using Biodegradable Sorbent Polymers Obtained from Polysaccharides
Alvarado, Sarah; Megía Fernández, Alicia; Ortega Muñoz, Mariano; Hernández Mateo, Fernando; López Jaramillo, Francisco Javier; Santoyo González, Francisco
Water use has been increasing globally by 1% per year, and recycling and re-use are critical issues compromised by the presence of pollutants. In this context, the design of novel materials and/or procedures for the large scale-removal of pollutants must be economically and environmentally feasible in order to be considered as part of the solution by emerging economies. We demonstrate that the cross-linking of biodegradable polysaccharides such as starch, dextrin, or dextrin and & beta;-cyclodextrin with divinyl sulfone is an innovative strategy for synthesizing insoluble and eco-friendly sorbent polymers, including pSt, pDx and pCD-Dx. The evaluation of these polymers' ability to remove ciprofloxacin (CIP), a prime example of antibiotic pollution, revealed that pSt, with a Kd of 1469 L/kg and a removal rate higher than 92%, is a favorable material. Its sorption is pH-dependent and enhanced at a mildly alkaline pH, allowing for the desorption (i.e., cleaning) and reuse of pSt through an environmentally friendly treatment with 20 mM AcONa pH 4.6. The facts that pSt (i) shows a high affinity for CIP even at high NaCl concentrations, (ii) can be obtained from affordable starting materials, and (iii) is synthesized and regenerated through organic, solvent-free procedures make pSt a novel sustainable material for inland water and seawater remediation, especially in less developed countries, due to its simplicity and low cost.
This research was funded by Junta de Andalucia through the research project B-FQM-316-UGR20 founded by the Programa Operativo FEDER Andalucia.; Supplementary Materials: The following supporting information can be downloaded at: https://www.
mdpi.com/article/10.3390/polym15153188/s1
Extracellular vesicles of Trypanosoma cruzi and immune complexes they form with sialylated and non-sialylated IgGs increase small peritoneal macrophage subpopulation and elicit different cytokines profilesCornet Gómez, AlbertoRetana Moreira, LissetteGómez Samblás, María MercedesOsuna Carrillo De Albornoz, Antoniohttps://hdl.handle.net/10481/845332023-09-21T07:49:33ZExtracellular vesicles of Trypanosoma cruzi and immune complexes they form with sialylated and non-sialylated IgGs increase small peritoneal macrophage subpopulation and elicit different cytokines profiles
Cornet Gómez, Alberto; Retana Moreira, Lissette; Gómez Samblás, María Mercedes; Osuna Carrillo De Albornoz, Antonio
American trypanosomiasis, or Chagas disease, is caused by the protozoan parasite Trypanosoma cruzi and is characterized by the presence of cardiac or gastrointestinal symptoms in a large number of patients during the chronic phase of the disease. Although the origin of the symptoms is not clear, several mechanisms have been described involving factors related to T. cruzi and the host immune response. In this sense, the extracellular vesicles (EVs) secreted by the parasite and the immune complexes (ICs) formed after their recognition by host IgGs (EVs-IgGs) may play an important role in the immune response during infection. The aim of the present work is to elucidate the modulation of the immune response exerted by EVs and the ICs they form by analyzing the variation in the subpopulations of small and large peritoneal macrophages after intraperitoneal inoculation in mice and to evaluate the role of the sialylation of the host IgGs in this immunomodulation. Both macrophage subpopulations were purified and subjected to cytokine expression analysis by RT-qPCR. The results showed an increase in the small peritoneal macrophage subpopulation after intraperitoneal injection of parasite EVs, but a greater increase in this subpopulation was observed when sialylated and non-sialylated ICs were injected, which was similar to inoculation with the trypomastigote stage of the parasite. The cytokine expression results showed the ability of both subpopulations to express inflammatory and non-inflammatory cytokines. These results suggest the role of free EVs in the acute phase of the disease and the possible role of immune complexes in the immune response in the chronic phase of the disease, when the levels of antibodies against the parasite allow the formation of immune complexes. The differential expression of interleukins showed after the inoculation of immune complexes formed with sialylated and non-sialylated IgGs and the interleukins expression induced by EVs, demonstrates that the IgG glycosilation is involved in the type of immune response that dominates in each of the phases of the 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; Fundacioin Ramoin 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)". Ministerio de Ciencia y Tecnologia of the Government of Spain funded the project PGC2018-099424-B-I00)". Exovesiculas circulantes como marcadoras de diagnostico, PREcoz de la Enfermedad de CHAGas Fundacion Ramon Areces 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 Tecnologiia of the government of Spain funded the project PGC2018-099424-B-I00.; The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fimmu.2023.1215913/full#supplementary-material