Expression of key steroidogenic enzymes in developing brain: hormonal compensation of sex chromosomes-induced sex differences

Developing brain of mammals is organized by gonadal steroids during the critical period of sexual differentiation (E18-PN10). The regulatory role of neurosteroids in the early brain is unclear. It is known that 17-β-estradiol (E2) is produced within the brain itself primarily due to local aromatization of gonadal testosterone and also due to de novo synthesis from cholesterol. Little is known about the circulating and local levels of steroids in the embryonic brain. Thus, the use of animal models that dissociate the effect of gonadal sex from sex chromosomes heritage facilitates the study of organizational actions of gonadal steroids and neurosteroids in each sex.http://www.saneurociencias.org.ar/congreso-2014/Fil: Cisternas, Carla Daniela. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra Introducción a la Química y Física Biológicas A; Argentina.Fil: Cisternas, Carla Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina.Fil: Tomé, Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Cambiasso, María Julia. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Biología Celular B; Argentina.Fil: Cambiasso, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina.Bioquímica y Biología Molecular (ídem 3.1.10

Abolition of the sex difference in Ngn3 by estradiol is depending on sex chromosome complement

A growing body of evidences indicates that some sexually dimorphic traits cannot be solely explained as a result of gonadal steroid action during the critical period of brain masculinization (E18-PN10). Neurogenin 3 (Ngn3), a gene located in mouse chromosome 10 (MGI:893591), is involved in neuritogenesis and morphological differentiation of hippocampal neurons (Salama-Cohen et al., 2006). Recent works from our laboratory have shown the existence of sex difference in the neuritogenic transcription factor Ngn3 in hypothalamic neurons before brain masculization. Moreover 17β-estradiol (E2) abolishes this sex difference (Scerbo et al., 2014). The sex difference in Ngn3 in hypothalamic neurons is depending on sex chromosome complement (Scerbo et al., 2014). In order to study if cell-autonomous actions of sex chromosomes are involved in the effect of E2 on Ngn3, we evaluated Ngn3 mRNA in neuronal cultures.http://www.saneurociencias.org.ar/congreso-2014/Fil: Tomé, Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Cisternas, Carla Daniela. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra Introducción a la Química y Física Biológicas A; Argentina.Fil: Cisternas, Carla Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina.Fil: Scerbo Jaureguiberry, Maria Julia. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina.Fil: Scerbo Jaureguiberry, Maria Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina.Fil: Cambiasso, María Julia. Universidad Nacional de Córdoba. Facultad de Odontología. Cátedra de Biología Celular B; Argentina.Fil: Cambiasso, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina.Bioquímica y Biología Molecular (ídem 3.1.10

Experiencias y desafíos sobre Educación Superior inclusiva

Una de las mayores barreras que encontramos aún en todos los países en los que hay planes nacionales de inclusión en marcha está precisamente en los pre-juicios y pre-supuestos sobre las posibilidades -y derechos- de las personas con necesidades especiales, con discapacidad o con diversidad funcional. Los procesos de exclusión social, tanto desde un punto de vista micro-sistémico (por ejemplo, en su salón de clases, en grupos de amigos o desconocidos…) como macro-sistémico (dificultades incluso legales para acceder a un empleo) presentan numerosas analogías con experiencias por las que ya hemos pasado como sociedades con otros grupos humanos que han sido abiertamente y generalizadamente excluidos, como es el caso de las mujeres, la mayoría de las cuales ha adquirido el derecho al voto a lo largo de la primera mitad del siglo XX o la discriminación racial. Esta memoria viva en el imaginario de nuestras sociedades debiera ayudarnos a decodificar con acierto los interrogantes que el proceso vivo de inclusión social y educativa nos formula, porque muchas de las soluciones encontradas ahí guardan paralelismos como las que proponemos a algunos para este sector de la ciudadanía con discapacidades o necesidades especiales. Si deseamos que todo lo anterior se materialice en proyectos de inclusión para todas las personas con necesidades especiales hacen falta por lo menos tres elementos interactuando activamente entre ellos. En primer lugar, la existencia de una democracia lo suficientemente afianzada y la vigencia de un estado de derecho. No conozco países con regímenes dictatoriales o autoritarios que tengan o hayan tenido planes de inclusión dignos de tal nombre. Efectivamente, la verdadera democracia significa no solo el ejercicio de los derechos formales de la misma, sino –entre otras muchas cosas- la creación de oportunidades para que los niños con limitaciones puedan acudir a los establecimientos escolares a los que acuden sus compañeros de edad, o que los trabajadores con discapacidad tengan acceso a trabajar en empresas ordinarias, con apoyos, si es preciso. La inclusión habita en el corazón mismo de la democracia. Segundo, es precisa la existencia de una masa crítica en la sociedad tanto de personas individuales como de organizaciones ciudadanas comprometidas con las personas con necesidades especiales, sus familias y amigos. Y por último, la presencia de una universidad activa en estas cuestiones que contribuya a la formación de los profesionales, investigue los muchos procesos que involucra un proyecto tan complejo como el de la inclusión educativa y social y colabore con los gestores de los servicios para promover políticas basadas en evidencias y análisis científicos.https://www.celei.cl/wp-content/uploads/2017/01/Experiencias-y-Desaf%C3%ADos-sobre-Educaci%C3%B3n-Superior-Inclusiva.pd

Práticas Integrativas e Complementares em Saúde no enfrentamento do período de pandemia da Covid-19 por trabalhadores remoto

A pandemia da Covid-19 trouxe enfrentamentos e adaptações ao estilo de vida dos indivíduos, necessitando de estratégias para prevenção da doença, com o isolamento e distanciamento social, o que incluiu o trabalho remoto (TR) e adaptação da rotina e estrutura ocupacional. Esta nova dinâmica repercutiu em impacto biopsicossocial, diminuindo o rendimento durante o trabalho, e gerando agravos físicos, psicológicos e emocionais. Diante deste contexto, faz-se necessário investigar recursos que minimizem estes impactos. Investigou-se o uso das Práticas Integrativas e Complementares em Saúde – PICS como recurso de enfrentamento à pandemia da Covid-19 por trabalhadores em atividade remota. Tratou-se de um estudo transversal, realizado por meio da aplicação de um questionário, via ferramenta Google Forms, para indivíduos acima de 18 anos que /estiveram em atividades ocupacionais remotas por pelo menos 3 meses durante a pandemia da Covid-19. Participaram do estudo 186 indivíduos de 20 a 70 anos selecionados aleatoriamente por convite em redes sociais, sendo que estes deveriam preencher os critérios de inclusão e poderiam pertencer a diferentes setores de trabalho. Sobre o impacto da pandemia na saúde, a maioria (40,32%) sentiu de forma “razoável”, enquanto o impacto do TR sobre a saúde foi relatado por 37,63% como “não prejudicial”. 66,67% dos participantes não praticavam nenhuma PICS antes da pandemia. Destes, 20,91% iniciaram alguma prática durante o isolamento, 78,26% faziam mais de uma modalidade e 21,74% apenas uma. Os motivos relatados para o início da prática foram: dores e/ou lesões ortopédicas, ansiedade e estresse. As práticas mais realizadas foram: meditação (14,5%) e yoga (10,22%). Para quem iniciou alguma prática, a importância em relação à saúde, foi considerada muito importante para 65,22% e quando questionados sobre a utilização das PICS, como estratégia de enfrentamento da pandemia, foi considerado muito importante por 47,83%. Conclui-se que as PICS foram recursos considerados importantes para a saúde e procurados para o enfrentamento da pandemia Covid-19, por trabalhadores em TR, sendo meditação e yoga as terapias mais utilizadas

Time to Switch to Second-line Antiretroviral Therapy in Children With Human Immunodeficiency Virus in Europe and Thailand.

Background: Data on durability of first-line antiretroviral therapy (ART) in children with human immunodeficiency virus (HIV) are limited. We assessed time to switch to second-line therapy in 16 European countries and Thailand. Methods: Children aged <18 years initiating combination ART (≥2 nucleoside reverse transcriptase inhibitors [NRTIs] plus nonnucleoside reverse transcriptase inhibitor [NNRTI] or boosted protease inhibitor [PI]) were included. Switch to second-line was defined as (i) change across drug class (PI to NNRTI or vice versa) or within PI class plus change of ≥1 NRTI; (ii) change from single to dual PI; or (iii) addition of a new drug class. Cumulative incidence of switch was calculated with death and loss to follow-up as competing risks. Results: Of 3668 children included, median age at ART initiation was 6.1 (interquartile range (IQR), 1.7-10.5) years. Initial regimens were 32% PI based, 34% nevirapine (NVP) based, and 33% efavirenz based. Median duration of follow-up was 5.4 (IQR, 2.9-8.3) years. Cumulative incidence of switch at 5 years was 21% (95% confidence interval, 20%-23%), with significant regional variations. Median time to switch was 30 (IQR, 16-58) months; two-thirds of switches were related to treatment failure. In multivariable analysis, older age, severe immunosuppression and higher viral load (VL) at ART start, and NVP-based initial regimens were associated with increased risk of switch. Conclusions: One in 5 children switched to a second-line regimen by 5 years of ART, with two-thirds failure related. Advanced HIV, older age, and NVP-based regimens were associated with increased risk of switch

Sex chromosome complement determines sex differences in aromatase expression and regulation in the stria terminalis and anterior amygdala of the developing mouse brain

Aromatase, which converts testosterone in estradiol, is involved in the generation of brain sex dimorphisms. Here we used the "four core genotypes" mouse model, in which the effect of gonadal sex and sex chromosome complement is dissociated, to determine if sex chromosomes influence the expression of brain aromatase. The brain of 16 days old XY mouse embryos showed higher aromatase expression in the stria terminalis and the anterior amygdaloid area than the brain of XX embryos, independent of gonadal sex. Furthermore, estradiol or dihydrotestosterone increased aromatase expression in cultures of anterior amygdala neurons derived from XX embryos, but not in those derived from XY embryos. This effect was also independent of gonadal sex. The expression of other steroidogenic molecules, estrogen receptor-α and androgen receptor was not influenced by sex chromosomes. In conclusion, sex chromosomes determine sex dimorphisms in aromatase expression and regulation in the developing mouse brain.Fil: Cisternas, Carla Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina. Universidad Nacional de Córdoba. Facultad de Odontología; ArgentinaFil: Tomé, Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Caeiro, Ximena Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Dadam, Florencia Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Garcia Segura, Luis Miguel. Consejo Superior de Investigaciones Científicas; EspañaFil: Cambiasso, Maria Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina. Universidad Nacional de Córdoba. Facultad de Odontología; Argentin

Galectin-1 exerts inhibitory effects during DENV-1 infection

Dengue virus (DENV) is an enveloped RNA virus that is mosquito-transmitted and can infect a variety of immune and non-immune cells. Response to infection ranges from asymptomatic disease to a severe disorder known as dengue hemorrhagic fever. Despite efforts to control the disease, there are no effective treatments or vaccines. In our search for new antiviral compounds to combat infection by dengue virus type 1 (DENV-1), we investigated the role of galectin-1, a widely-expressed mammalian lectin with functions in cell-pathogen interactions and immunoregulatory properties. We found that DENV-1 infection of cells in vitro exhibited caused decreased expression of Gal-1 in several different human cell lines, suggesting that loss of Gal-1 is associated with virus production. In test of this hypothesis we found that exogenous addition of human recombinant Gal-1 (hrGal-1) inhibits the virus production in the three different cell types. This inhibitory effect was dependent on hrGal-1 dimerization and required its carbohydrate recognition domain. Importantly, the inhibition was specific for hrGal-1, since no effect was observed using recombinant human galectin-3. Interestingly, we found that hrGal-1 directly binds to dengue virus and acts, at least in part, during the early stages of DENV-1 infection, by inhibiting viral adsorption and its internalization to target cells. To test the in vivo role of Gal-1 in DENV infection, Gal-1-deficient-mice were used to demonstrate that the expression of endogenous Galectin-1 contributes to resistance of macrophages to in vitro-infection with DENV-1 and it is also important to physiological susceptibility of mice to in vivo infection with DENV-1. These results provide novel insights into the functions of Gal-1 in resistance to DENV infection and suggest that Gal-1 should be explored as a potential antiviral compound

Arctium lappa Extract Suppresses Inflammation and Inhibits Melanoma Progression

Background: Arctium lappa has been used as popular medicinal herb and health supplement in Chinese societies. Bioactive components from A. lappa have attracted the attention of researchers due to their promising therapeutic effects. In this study, we investigated the effects of A. lappa hydroalcoholic extract (Alhe) during different models of inflammation, in vivo. Methods: The anti-inflammatory activity was evaluated through the air pouch model. For this, mice received an inflammatory stimulus with lipopolysaccharide (LPS) and were later injected with Alhe. To assess anti-tumoral activity, the animals were inoculated with B16F10 cells and injected with Alhe every 5 days, along the course of 30 days. Controls were submitted to the same conditions and injected with the vehicle. Peritoneal or air pouch fluids were collected to evaluate leukocyte counting or cellular activation via quantification of cytokines and nitric oxide. Results: Alhe injection reduced the neutrophil influx and production of inflammatory mediators in inflammatory foci after LPS or tumor challenges. Furthermore, Alhe injection reduced tumor growth and enhanced mice survival. Conclusions: Collectively, these data suggest that Alhe regulates immune cell migration and activation, which correlates with favorable outcome in mouse models of acute inflammation and melanoma progression

Gal-1 acts at early stages during DENV-1 infection.

<p><b>(A)</b> Binding of DENV-1 to hrGal-1 in a dose-dependent manner. Serial two-fold dilutions of DENV-1 were applied to 96-well plates coated with 1 µg of hrGal-1 per well, and the bound virus particles were detected by ELISA with mouse anti-E protein antibody. BSA-coated wells served as the negative control. To assess the involvement of Gal-1 CRD, the assay was performed in presence of 40 mM lactose or sucrose. Each value represents the mean±the SD from 4 assays performed in duplicates. <b>(B)</b> Adsorption and internalization assays: for adsorption assay, ECV-304 cells were infected with DENV-1 at MOI of 10 in presence or absence of 10 µM hrGal-1 during 1 h at 4°C and then washed to remove viral inoculum. Cells were collected and the viral RNA was quantified by Real-Time PCR. Data was normalized by host β-actin expression. For internalization assay, ECV-304 cells were inoculated with DENV-1 (MOI of 10) at 4°C for 1 hour. Then, cells were washed and transferred to 37°C and hrGal-1 (10 µM) or only medium were added to culture. After 1 hour of incubation, non-internalized viruses were inactivated with citrate buffer and viral loads were quantified by Real-Time PCR. Data is presented as Viral RNA amount equivalents to PFU/mL±SD from 3 experiments assessed in triplicates. <b>(C)</b> For virucidal assay, DENV-1 was incubated with hrGal-1, in the presence or absence of RNAse. After 1 h incubation at 37°C, RNA was isolated and subjected to RT-Real-Time PCR. Purified viral RNA incubated or not with RNAse was used as control (N = 3). <b>(D)</b> ECV-304 cells were infected with DENV-1 at a MOI of 0.5 (DENV-1). For the treatments, hrGal-1 was incubated with ECV-304 and DENV-1 simultaneously (ECV+Gal-1+DENV), or hrGal-1 (10 µM) was pre-incubated with either ECV-304 cells or with DENV-1 (MOI 0.5) for 60 minutes before the inoculation (ECV+hrGal-1)+DENV versus (DENV+hrGal-1)+ECV, respectively. At 72 hours postinfection, supernatants were collected and the viral loads were quantified by Real-Time PCR (N = 3). **p<0.001; ***p<0.0001.</p

Treatment with human recombinant Gal-1 inhibits DENV-1 in vitro infection.

<p><b>(A)</b> Biotinylated-hrGal-1 (20 µg/mL) was incubated with ECV-304 cells in presence or absence of 40 mM lactose or sucrose for 1 hour at 4°C. The binding of biotinylated-hrGal-1 to ECV-304 cells surfaces was detected by staining with streptavidin-FITC and measured by flow cytometry. The analysis was performed using a Diva software (Becton Dickson) and results are expressed as mean mean fluorescence intensity (MFI)±SD. Tests were performed in triplicates. <b>(B)</b> HMVEC-L, Vero-E6 and ECV-304 cells were incubated with 10 µM hrGal-1 or only medium for 1 hour at 37°C. Following, cells were inoculated with DENV-1 at a MOI of 0.5 and cultivated for 72 hours. At 72 hours postinfection the supernatants were collected and the viral RNA amounts were quantified by Real-Time PCR. Results are shown as Viral RNA amounts equivalent to PFU/ml±SD from 3 assays performed in triplicates. <b>(C)</b> ECV-304 cells were treated with hrGal-1 and infected with DENV-1 as described in (B) for 120 hours. The supernatants were collected at the indicated times postinfection and the viral loads were quantified by Real-Time PCR as described in (B). (N = 3) <b>(D)</b> ECV-304 cells were incubated with increased concentrations of monomeric-Gal-1 (hrGal-1 m), dimeric-Gal-1 (hrGal-1 d) or galectin-3 (hrGal-3) for 1 hour at 37°C before inoculation with DENV-1 (MOI 0.5). Cells were cultivated for 72 hours at 37°C and viral load was quantified as described in (A) (N = 3). <b>(E)</b> ECV-304 cells were treated with hrGal-1 (10 µM) in the presence of 40 mM lactose (LAC) or 40 mM sucrose (SUC) and then infected with DENV-1 as described in (B), for 72 hours. The viral load was quantified as described in (B). Data are representative from three independent experiments. *p<0.01; **p<0.001; ***p<0.0001.</p