Upregulation of interferon-alpha gene in bovine embryos produced in vitro in response to experimental infection with noncytophatic bovine-viral-diarrhea virus

In-vitro fertilization is a routine livestock-breeding technique widely used around the world. Several studies have reported the interaction of bovine viral-diarrhea virus (BVDV) with gametes and in-vitro-produced (IVP) bovine embryos. Since, gene expression in BVDV-infected IVP bovine embryos is scarcely addressed. The aim of this work was to evaluate the differential expression of genes involved in immune and inflammatory response. Groups of 20–25 embryos on Day 6 (morula stage) were exposed (infected) or not (control) to an NCP-BVDV strain in SOF medium. After 24 h, embryos that reached expanded blastocyst stage were washed. Total RNA of each embryo group was extracted to determine the transcription levels of 9 specific transcripts related with antiviral and inflammatory response by SYBR Green real time quantitative (RT-qPCR). Culture media and an aliquot of the last embryos wash on Day 7 were analyzed by titration and virus isolation, respectively. A conventional PCR confirmed BVDV presence in IVP embryos. A significantly higher expression of interferon-α was observed in blastocysts exposed to NCP-BVDV compared to the controls (p < 0.05). In this study, the upregulation of INFα and TLR7 genes involved in inflammatory and immune response in BVDV-infected IVP bovine embryos is a new finding in this field. This differential expression suggest that embryonic cells could function in a manner like immune cells by recognizing and responding early to interaction with viral pathogens. These results provide new insights into the action of BVDV on the complex molecular pathways controlling bovine early embryonic development.Fil: Gonzalez Altamiranda, Erika Analia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Buenos Aires Sur. Estacion Experimental Agropecuaria Balcarce. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible.; ArgentinaFil: Arias, María E.. Universidad de La Frontera. Núcleo Científico y Tecnológico en Recursos Naturales; ChileFil: Kaiser, Germán Gustavo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Buenos Aires Sur. Estacion Experimental Agropecuaria Balcarce. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible.; ArgentinaFil: Mucci, Nicolás Crescencio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mar del Plata. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Buenos Aires Sur. Estacion Experimental Agropecuaria Balcarce. Instituto de Innovación Para la Producción Agropecuaria y El Desarrollo Sostenible.; ArgentinaFil: Odeón, Anselmo C.. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; ArgentinaFil: Felmer Dörner, Ricardo Nicolás. Universidad de La Frontera. Núcleo Científico y Tecnológico en Recursos Naturales; Chil

Reliability of enzyme assays in dried blood spots for diagnosis of 4 lysosomal storage disorders

Introduction: Lysosomal storage disorders (LSD) are inherited diseases caused, in the majority of them, by the deficiency of lysosomal enzymatic activities. Ob-jectives: We aimed to analyze the usefulness of DBS samples for diagnosis of 4 LSDs, with the availability of a large quantity of patient samples. Design and methods: Blood samples from previously diagnosed patients with Fabry, Gaucher, Hunter, and Maro-teaux-Lamy syndromes and normal control indi-viduals, were collected and dispen-sed in filter paper, and used for enzymatic activity determination. Re-sults: Diagnosis of hemi/homo-zygous patients with Fabry, Hunter and Maroteaux-Lamy diseases using DBS samples showed ideal parameters of 100% sen-sitivity and specificity. DBS assay for Gaucher dis-ease would need a posterior confirmatory step. Con-clusions: Leukocyte measu-rement is the only reli-able way to diagnose Gaucher disease. For Hunter, Fabry and Maroteaux-Lamy disorders discrimina-tion between patients and controls seems adequate by DBS.Laboratorio de Investigaciones del Sistema Inmun

Alteraciones en proteínas asociadas al manejo del calcio intracelular en corazón del modelo murino de enfermedad de Fabry

La Enfermedad de Fabry es una patología de almacenamiento lisosomal genética que entre varios órganos, afecta también al tejido cardíaco. Resultados previos de nuestro grupo revelaron menor contractilidad del ventrículo izquierdo, de músculos papilares y de miocitos aislados, asociado a menor amplitud del transitorio de calcio (Ca2+) en el modelo murino de Enfermedad de Fabry (knockout para el gen de alfagalactosidasa A) (RF) en comparación con los de la cepa salvaje (RWT). El objetivo del presente trabajo es analizar el estado de las proteínas asociadas a la homeostasis del Ca2+ miocárdico en corazones de ratones RF y RWT.Facultad de Ciencias Médica

Evaluación de la disfunción cardíaca en el modelo murino de enfermedad de Fabry

Los objetivos del presente trabajo son analizar la función cardíaca mediante ecocardiograma y la actividad contráctil de músculos papilares aislados y cardiomiocitos del corazón de ratones Fabry (RF) en comparación con los de cepa salvaje (RWT).Facultad de Ciencias Médica

Box d'estudi i passadís de la zona de silenci de la Biblioteca del Campus del Baix Llobregat

Fotografies realitzades per encàrrec del Servei de Biblioteques, Publicacions i Arxius per tal d'il·lustrar l'ús del servei i les biblioteques de la universitat. I per tal de poder usar les imatges en diferents àmbits: difusió de la institució, memòries i altres publicacions

Diet effects on female reproduction in high growth (hg/hg) mice that are deficient in the Socs-2 gene

The detrimental effect of larger body size on reproductive performance has been well documented in mouse models of overgrowth, such as Growth Hormone (GH) transgenics. This study describes the reproductive performance of the High Growth (HG) mouse model of overgrowth. The HG mouse model exhibits overgrowth due to a partially recessive autosomal mutation that increases growth rate and mature body size. The HG phenotype results from the lack of expression of Socs-2, which negatively regulates GH signaling. C57BL/6J (C57) and congenic C57BL/6J-hg/hg (HG) female mice were fed four diets differing in protein and energy content, starting at 8 weeks of age. A complete reproductive cycle from mating to weaning was evaluated. HG mice were 40% larger than C57 and had a higher feed intake throughout the experiment. Significant genotype × diet interactions were detected for growth, body composition and reproductive traits. HG females showed poor reproductive performance compared to controls as demonstrated by their lower fertility during mating, which was not overcome by changes in the diet. No differences were detected in litter size, but HG animals exhibited a longer gestation length and heavier pup weaning weights compared to controls. Reproductive impairment in HG seems to be the consequence of the lack of Socs-2 independent of the effects of increased body size on reproduction

Higher apoptotic state in Fabry disease peripheral blood mononuclear cells: effect of globotriaosylceramide

Fabry disease is an X-linked lysosomal storage disorder (LSD) due to deficiency of the enzyme α-galactosidase A, resulting in intracellular deposition of globotriaosylceramide (Gb3). Accumulation of Gb3 is probably related to tissue and organ dysfunctions. Diverse pathological mechanisms are elicited in LSDs, giving together the phenotypic expression of each disease. The purpose of the present study is to investigate if apoptosis could play a role in Fabry disease pathogenesis and to understand the mechanisms involved in the proapoptotic state. We have demonstrated that Fabry disease peripheral blood mononuclear cells display a higher apoptotic state, which is reduced by enzyme replacement therapy (ERT), and is mediated, at least in part, by activation of the intrinsic pathway of caspases. We could rule out the implication of “unfolded protein response-ER stress” in this apoptotic process. To further confirm the suggestion that Gb3 is associated to apoptotic cell death, we treated normal cells with Gb3 at concentrations found in Fabry patients. Addition of Gb3 resulted in a dose-dependent induction of apoptosis involving the intrinsic pathway. In summary, PBMC from Fabry patients display a higher apoptotic state, which could be mainly related to elevated Gb3.Fil: de Francesco, Pablo Nicolás. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mucci, Juan Marcos. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ceci, Romina. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; ArgentinaFil: Fossati, Carlos Alberto. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rozenfeld, Paula Adriana. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.

Reproductive biotechnologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA) of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70), endoplasmic reticulum (ER) stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5) and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3) in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A) + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip) was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART)

Induction of osteoclastogenesis in an in vitro model of Gaucher disease is mediated by T cells via TNF-α

Gaucher disease is a lysosomal storage disorder caused by deficiency of glucocerebrosidase enzymatic activity leading to accumulation of its substrate glucocerebrosidase mainly in macrophages. Skeletal disorder of Gaucher disease is the major cause of morbidity and is highly refractory to enzyme replacement therapy. However, pathological mechanisms of bone alterations in Gaucher disease are still poorly understood. We hypothesized that cellular alteration in Gaucher disease produces a proinflammatory milieu leading to bone destruction through enhancement of monocyte differentiation to osteoclasts and osteoclasts resorption activity. Against this background we decided to investigate in an in vitro chemical model of Gaucher disease, the capacity of secreted soluble mediators to induce osteoclastogenesis, and the mechanism responsible for this phenomena. We demonstrated that soluble factors produced by CBE-treated PBMC induced differentiation of osteoclasts precursors into mature and active osteoclasts that express chitotriosidase and secrete proinflammatory cytokines. We also showed a role of TNF-α in promoting osteoclastogenesis in Gaucher disease chemical model. To analyze the biological relevance of T cells in osteoclastogenesis of Gaucher disease, we investigated this process in T cell-depleted PBMC cultures. The findings suggest that T cells play a role in osteoclast formation in Gaucher disease. In conclusion, our data suggests that in vitro GCASE deficiency, along with concomitant glucosylceramide accumulation, generates a state of osteoclastogenesis mediated in part by pro-resorptive cytokines, especially TNF-α. Moreover, T cells are involved in osteoclastogenesis in Gaucher disease chemical model.Fil: Mucci, Juan Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; ArgentinaFil: Scian, Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: de Francesco, Pablo Nicolás. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Suqueli García, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; ArgentinaFil: Ceci, Romina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; ArgentinaFil: Fossati, Carlos Alberto. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Delpino, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Estudios de la Inmunidad Humoral Prof. Ricardo A. Margni; ArgentinaFil: Rozenfeld, Paula Adriana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigaciones del Sistema Inmune; Argentin