Fyn Mediates Leptin Actions in the Thymus of Rodents

BACKGROUND:Several effects of leptin in the immune system rely on its capacity to modulate cytokine expression and apoptosis in the thymus. Surprisingly, some of these effects are dependent on signal transduction through the IRS1/PI3-kinase, but not on the activation of JAK2. Since all the well known effects of leptin in different cell types and tissues seem to be dependent on JAK2 activation, we hypothesized that, at least for the control of thymic function, another, unknown kinase could mediate the transduction of the leptin signal from the ObR towards the IRS1/PI3-kinase signaling cascade. METHODOLOGY/PRINCIPAL FINDINGS:Here, by employing immunoblot, real-time PCR and flow citometry we show that the tyrosine kinase, Fyn, is constitutively associated with the ObR in thymic cells. Following a leptin stimulus, Fyn undergoes an activating tyrosine phosphorylation and a transient association with IRS1. All these effects are independent of JAK2 activation and, upon Fyn inhibition, the signal transduction towards IRS1/PI3-kinase is abolished. In addition, the inhibition of Fyn significantly modifies the effects of leptin on thymic cytokine expression. CONCLUSION/SIGNIFICANCE:Therefore, in the thymus, Fyn acts as a tyrosine kinase that transduces the leptin signal independently of JAK2 activation, and mediates some of the immunomodulatory effects of leptin in this tissue

Detection of epithelial apoptosis in pelvic ileal pouches for ulcerative colitis and familial adenomatous polyposis

<p>Abstract</p> <p>Background</p> <p>Ileal pouch-anal anastomosis (IPAA) is the surgical procedure of choice for patients with refractory ulcerative colitis (UC) and for familial adenomatous polyposis (FAP) with many rectal polyps. Pouchitis is one of the more frequent complications after IPAA in UC patients; however, it is rare in FAP.</p> <p>Objective</p> <p>Evaluate pro-apoptotic activity in endoscopically and histological normal mucosa of the ileal pouch in patients with UC and FAP.</p> <p>Methods</p> <p>Eighteen patients (nine with UC and nine with FAP) with J pouch after total rectocolectomy were studied. Biopsies were obtained from the mucosa of the pouch and from normal ileum. The specimens were snap-frozen and the expressions of Bax and Bcl-2 were determined by immunoblot of protein extracts and by immunohistochemistry analysis. FADD, Caspase-8, APAF-1 and Caspase-9 were evaluated by immunoprecipitation and immunoblot.</p> <p>Results</p> <p>Patients with UC had significantly higher protein levels of Bax and APAF-1, Caspase-9 than patients with FAP, but were similar to controls. The expressions of Bcl-2 and FADD, Caspase-8 were similar in the groups. Immunohistochemistry for Bax showed less intensity of immunoreactions in FAP than in UC and Controls. Bcl-2 immunostaining was similar among the groups.</p> <p>Conclusion</p> <p>Patients with FAP present lower levels of pro-apoptotic proteins in all methods applied, even in the absence of clinical and endoscopic pouchitis and dysplasia in the histological analysis. These findings may explain a tendency of up-regulation of apoptosis in UC patients, resulting in higher rates of progression to pouchitis in these patients, which could correlate with mucosal atrophy that occurs in inflamed tissue. However, FAP patients had low pro-apoptotic activity in the mucosa, and it could explain the tendency to low cell turn over and presence of adenomas in this syndrome.</p

Unsaturated Fatty Acids Revert Diet-Induced Hypothalamic Inflammation in Obesity

Background: In experimental models, hypothalamic inflammation is an early and determining factor in the installation and progression of obesity. Pharmacological and gene-based approaches have proven efficient in restraining inflammation and correcting the obese phenotypes. However, the role of nutrients in the modulation of hypothalamic inflammation is unknown. Methodology/Principal Findings: Here we show that, in a mouse model of diet-induced obesity, partial substitution of the fatty acid component of the diet by flax seed oil (rich in C18:3) or olive oil (rich in C18:1) corrects hypothalamic inflammation, hypothalamic and whole body insulin resistance, and body adiposity. In addition, upon icv injection in obese rats, both v3 and v9 pure fatty acids reduce spontaneous food intake and body mass gain. These effects are accompanied by the reversal of functional and molecular hypothalamic resistance to leptin/insulin and increased POMC and CART expressions. In addition, both, v3 and v9 fatty acids inhibit the AMPK/ACC pathway and increase CPT1 and SCD1 expression in the hypothalamus. Finally, acute hypothalamic injection of v3 and v9 fatty acids activate signal transduction through the recently identified GPR120 unsaturated fatty acid receptor. Conclusions/Significance: Unsaturated fatty acids can act either as nutrients or directly in the hypothalamus, reverting dietinduced inflammation and reducing body adiposity. These data show that, in addition to pharmacological and geneti

Trajetória profissional de egressos de cursos de doutorado nas áreas da saúde e biociências

OBJETIVO: Analisar a trajetória e satisfação profissional de egressos de cursos de doutorado na área da saúde. MÉTODOS: Estudo exploratório com 827 egressos dos cursos de doutorado da Fundação Oswaldo Cruz nas áreas da saúde coletiva, biociências e atenção à saúde, entre 1984 e 2007. Os sujeitos foram agrupados em três coortes temporais. Foi analisado o perfil dos egressos; mapeadas suas trajetórias profissionais, suas percepções sobre a formação recebida; suas motivações para escolha da instituição para realizar o doutorado; e as avaliações efetuadas sobre os cursos. Utilizou-se questionário em formato eletrônico para preenchimento on-line para coleta de dados. RESULTADOS: Existiram diferenças entre as coortes de egressos, relacionadas ao período em que cursaram o doutorado, a distintas trajetórias de formação e processos de trabalho entre egressos das áreas de biociências e saúde e as peculiaridades das diferentes áreas em que a instituição oferece cursos de doutorado: saúde coletiva, biociências e atenção à saúde. CONCLUSÕES: Os resultados possibilitam ampliar o conhecimento das instâncias de gestão acadêmica sobre os processos de formação, estabelecendo uma “linha de base” para o acompanhamento da trajetória dos egressos e contribuir com subsídios para o aprimoramento dos processos de acompanhamento de egressos dos programas de pós-graduaçãoOBJECTIVE: To analyze the career path and professional satisfaction of alumni from the doctorate degree programs in health sector. METHODS: Exploratory study with 827 alumni of doctoral programs in public health, biological and health sciences at the Fundação Oswaldo Cruz, RJ, Southeastern Brazil, from 1984 to 2007. The subjects were grouped in three cross-temporal cohorts according to year. The profiles of the alumni were analyzed, their career paths mapped and information on the perceptions of the education they received and the reasons that led them to choose the institute for their doctoral courses gathered, as well as their evaluations of the courses. The data were collected by means of an online questionnaire. RESULTS: There are differences between cohorts of alumni related to the periods they followed the courses, their distinct educational backgrounds and labor processes between those from the biological and health sciences areas, and to the specificities of the different areas where the institution offers doctoral courses: public health, biological and health sciences. CONCLUSIONS: The results allow the academic management of the educational processes to expend its knowledge, thus establishing a baseline for tracking the trajectory of alumni, and may contribute to upgrading the follow up process of Brazilian graduate program

Central Exercise Action Increases the AMPK and mTOR Response to Leptin

AMP-activated protein kinase (AMPK) and mammalian Target of Rapamycin (mTOR) are key regulators of cellular energy balance and of the effects of leptin on food intake. Acute exercise is associated with increased sensitivity to the effects of leptin on food intake in an IL-6-dependent manner. To determine whether exercise ameliorates the AMPK and mTOR response to leptin in the hypothalamus in an IL-6-dependent manner, rats performed two 3-h exercise bouts, separated by one 45-min rest period. Intracerebroventricular IL-6 infusion reduced food intake and pretreatment with AMPK activators and mTOR inhibitor prevented IL-6-induced anorexia. Activators of AMPK and fasting increased food intake in control rats to a greater extent than that observed in exercised ones, whereas inhibitor of AMPK had the opposite effect. Furthermore, the reduction of AMPK and ACC phosphorylation and increase in phosphorylation of proteins involved in mTOR signal transduction, observed in the hypothalamus after leptin infusion, were more pronounced in both lean and diet-induced obesity rats after acute exercise. Treatment with leptin reduced food intake in exercised rats that were pretreated with vehicle, although no increase in responsiveness to leptin-induced anorexia after pretreatment with anti-IL6 antibody, AICAR or Rapamycin was detected. Thus, the effects of leptin on the AMPK/mTOR pathway, potentiated by acute exercise, may contribute to appetite suppressive actions in the hypothalamus

The Secret Life of the Anthrax Agent Bacillus anthracis: Bacteriophage-Mediated Ecological Adaptations

Ecological and genetic factors that govern the occurrence and persistence of anthrax reservoirs in the environment are obscure. A central tenet, based on limited and often conflicting studies, has long held that growing or vegetative forms of Bacillus anthracis survive poorly outside the mammalian host and must sporulate to survive in the environment. Here, we present evidence of a more dynamic lifecycle, whereby interactions with bacterial viruses, or bacteriophages, elicit phenotypic alterations in B. anthracis and the emergence of infected derivatives, or lysogens, with dramatically altered survival capabilities. Using both laboratory and environmental B. anthracis strains, we show that lysogeny can block or promote sporulation depending on the phage, induce exopolysaccharide expression and biofilm formation, and enable the long-term colonization of both an artificial soil environment and the intestinal tract of the invertebrate redworm, Eisenia fetida. All of the B. anthracis lysogens existed in a pseudolysogenic-like state in both the soil and worm gut, shedding phages that could in turn infect non-lysogenic B. anthracis recipients and confer survival phenotypes in those environments. Finally, the mechanism behind several phenotypic changes was found to require phage-encoded bacterial sigma factors and the expression of at least one host-encoded protein predicted to be involved in the colonization of invertebrate intestines. The results here demonstrate that during its environmental phase, bacteriophages provide B. anthracis with alternatives to sporulation that involve the activation of soil-survival and endosymbiotic capabilities

Phenotypic changes in low-density lipoprotein particles as markers of adverse clinical outcomes in COVID-19

Background and aims: Low-density lipoprotein (LDL) plasma concentration decline is a biomarker for acute inflammatory diseases, including coronavirus disease-2019 (COVID-19). Phenotypic changes in LDL during COVID-19 may be equally related to adverse clinical outcomes. Methods: Individuals hospitalized due to COVID-19 (n = 40) were enrolled. Blood samples were collected on days 0, 2, 4, 6, and 30 (D0, D2, D4, D6, and D30). Oxidized LDL (ox-LDL), and lipoprotein-associated phospholipase A2 (Lp-PLA2) activity were measured. In a consecutive series of cases (n = 13), LDL was isolated by gradient ultracentrifugation from D0 and D6 and was quantified by lipidomic analysis. Association between clinical outcomes and LDL phenotypic changes was investigated. Results: In the first 30 days, 42.5% of participants died due to Covid-19. The serum ox-LDL increased from D0 to D6 (p < 0.005) and decreased at D30. Moreover, individuals who had an ox-LDL increase from D0 to D6 to over the 90th percentile died. The plasma Lp-PLA2 activity also increased progressively from D0 to D30 (p < 0.005), and the change from D0 to D6 in Lp-PLA2 and ox-LDL were positively correlated (r = 0.65, p < 0.0001). An exploratory untargeted lipidomic analysis uncovered 308 individual lipids in isolated LDL particles. Paired-test analysis from D0 and D6 revealed higher concentrations of 32 lipid species during disease progression, mainly represented by lysophosphatidyl choline and phosphatidylinositol. In addition, 69 lipid species were exclusively modulated in the LDL particles from non-survivors as compared to survivors. Conclusions: Phenotypic changes in LDL particles are associated with disease progression and adverse clinical outcomes in COVID-19 patients and could serve as a potential prognostic biomarker

Abundance and stratification of soil macroarthropods in a Caatinga Forest in Northeast Brazil

In arid and semiarid environments, seasonality usually exerts a strong influence on the composition and dynamics of the soil community. The soil macroarthropods were studied in a Caatinga forest located in the Reserva Particular do Patrimônio Natural (RPPN) Fazenda Almas, São José dos Cordeiros, Paraíba, Brazil. Samples were collected during the dry and rainy seasons following the method proposed by the Tropical Soil Biology and Fertility Program (TSBF), with minor modifications. At each station, 15 soil blocks (20 × 20 × 30 cm: 12 L) were extracted and divided into three layers: A (0-10 cm), B (10-20 cm), and C (20-30 cm). In the rainy and dry seasons 1,306 ± 543(se) and 458 ± 212 ind.m-2 macroarthropods were found, respectively, with 35 and 18 respective taxa recorded. The abundance of individuals and taxa were significantly higher in the rainy season. Isoptera (57.8%) was the most abundant taxon, followed by Hymenoptera: Formicidae (17.2%), Coleoptera larvae (7.3%), and Araneae (3.5%). In the rainy season, abundance in layer A (576 ± 138 ind.m-2) was significantly higher than that of layer C (117 ± 64 ind.m-2), but was not different from layer B (613 ± 480 ind.m-2). There was also no difference between the layer B and C abundances. In the dry season, abundance in layer B (232 ± 120 ind.m-2) was not significantly different compared to layer A (182 ± 129 ind.m-2), but was significantly higher than abundance in layer C (44 ± 35 ind.m-2). During the rainy season, layer A (34 taxa) was significantly richer in taxa than layers B (19 taxa) and C (11 taxa). On the other hand, during the dry season the richness of layers A (12 taxa) and B (12 taxa) was equal, but significantly higher than that of layer C (6 taxa). Richness of taxa and abundance were positively correlated with soil organic matter and negatively correlated with soil temperature. The community of soil macroarthropods in the area of Caatinga studied has taxonomic and functional structures that are relatively complex and is therefore likely to exert an influence on ecosystem productivity due to its physical effects on soil profile and necromass fragmentation, as occurs in other arid and semiarid ecosystems throughout the world