Genomic characterization and seroprevalence studies on alphaviruses in Uruguay

Alphaviruses (Togaviridae) are arboviruses frequently associated with emerging infectious diseases. In this study, we aimed to investigate the presence of alphaviruses in Uruguay by detecting the viral genome in mosquitoes and neutralizing antibodies in equines. A total of 3,575 mosquitoes were analyzed for alphavirus genome detection. Serologic studies were performed on 425 horse sera by plaque reduction neutralization test (PRNT80) against Venezuelan equine encephalitis virus (VEEV) subtype IAB, Pixuna virus (PIXV), Rio Negro virus (RNV), western equine encephalitis virus (WEEV), and Madariaga virus (MADV). Mosquitoes belonging to six genera were captured and 82.9% were identified as Culex pipiens. Two Cx. pipiens pools collected in Fray Bentos and Las Toscas localities were alphavirus positive, and phylogenetic analyses showed that the sequences grouped into two different clusters: the lineage I of eastern equine encephalitis virus and RNV (VEEV complex), respectively. Plaque reduction neutralization test assays showed antibodies against strains of the VEEV complex, MADV, and WEEV. Rio Negro virus was the most geographically widespread virus, showing higher seroprevalences (up to 20%). Seroprevalences against VEEV IAB ranged between 4.6% and 13%; antibodies against PIXV, WEEV, and MADV were less frequent (3–4%). In conclusion, RNV exhibited the highest seroprevalence in horses, a wide geographical distribution, and viral genome was detected in Cx. pipiens mosquitoes. Madariaga virus had a low seroprevalence in equines, but an epizootic lineage typical of North America was detected in Cx. pipiens mosquitoes. Taken together, our results show that alphaviruses are present in Uruguay with variable occurrence and geographical distribution being a potential threat for human and equine health

Prenatal stress and later metabolic consequences : systematic review and meta-analysis in rodents

Abstract: Background: Numerous rodent studies have evaluated the effects of maternal stress (MS) on later in life susceptibility to Metabolic Syndrome (MetS) intermediate phenotypes with varying results. The aim of this study was to quantitatively synthesize the available data on the effects of MS on offspring obesity, estimated indirectly by body mass (BM), body fat (BF) and plasma leptin; systolic blood pressure (SBP); plasma glucose (and insulin) and blood lipid concentrations. Methods: Literature was screened and summary estimates of the effect of MS outcomes were calculated by using random-effects models. Data on the effects of exogenous corticosteroid administration (or inhibition of 11β-HSD2) during pregnancy in rodents was analysed separately to characterize the direct phenotypic effects of prenatal corticosteroid excess (PCE). Results: We conducted 14 separate meta-analyses and synthesized relevant data on outcomes scarcely reported in literature. Both MS and PCE were associated with low birth weight without rapid catch-up growth resulting in decreased body mass later in life. Our analysis also revealed significant and contradictory effects on offspring adiposity. Little evidence was found for effects on glucose metabolism and blood lipids. We identified increased SBP in offspring exposed to PCE; however, there is not enough data to draw any conclusion about effects of MS on SBP. Conclusions: Neonatal weight proved to be decreased in offspring prenatally exposed to stress or corticosteroids, but laboratory rodents in the absence of a challenging environment did not show catch-up growth. The available evidence is inconclusive regarding the effect on adiposity revealing clear methodological and knowledge gaps. This meta-analysis also confirmed a significant positive association between PCE and SBP. Nevertheless, additional studies should address the association with MS

Systematic review and meta-analysis on the relationship between prenatal stress and metabolic syndrome intermediate phenotypes

BACKGROUND: Metabolic Syndrome (MetS) can be considered as a consequence of a complex interplay between genetic and environmental factors and can be influenced by changes in the environment early in life. Prenatal stress (PS) exposure likely represents an important adverse intrauterine environment that may impact the biology of the developing organism. The aim of this study was to quantitatively synthesize the available data on the effects of PS on offspring's obesity, estimated indirectly by body mass index (BMI) and body fat; blood pressure, plasma glucose and blood lipid concentrations (triglycerides and high-density lipoprotein cholesterol). METHODS: Literature searches for eligible studies on PubMed were conducted until October 8, 2018. Full text review yielded 24 publications for inclusion into the systematic review. Meta-analyses were performed for the outcomes BMI and body fat. 62 effect sizes from 19 studies together with relevant moderators were collected. Summary estimates were calculated by using random-effects model. RESULTS: The combined standardized mean difference (d) for the relation between BMI and PS indicated that despite significant heterogeneity, stress exposure of expectant mothers was associated with increased BMI of their offspring [d (95% CI) = 0.268 (0.191; 0.345)]. Both objective and subjective stress have been linked to increased overweight. Preliminary results of the relationship between PS and body fat suggested that the contribution of PS to body fat should be at least further considered [d (95% CI) = 0.167 (0.016; 0.317)]. Evidence from a limited number of published studies do not sustains an effect on blood pressure, glucose metabolism or circulating lipids, however these outcomes have only been scarcely investigated. CONCLUSIONS: A direct association between PS and BMI was found and further studies are needed to confirm the relationship between maternal stress during gestation and body fat. Overall, findings suggest that PS could contribute to alterations to the post-natal offspring phenotype

Effects of prenatal stress and postnatal high fat diet feeding on BALB/c mice metabolism

Resumen: In-utero exposure to maternal stress increases short and long term risk of suffering metabolic diseases. Exposure to stressful events leads to an increase in glucocorticoids release by activation of the HPA axis, therefore early programming of the HPA axis has emerged as a key underlying mechanism of stress-related disorders. Evidence suggests that a stressful prenatal environment seems to favour adverse metabolic conditions. To test this hypothesis in BALB/c mice, a strain susceptible to stress but resistant to metabolic effects of a high fat diet (HFD), we exposed female pregnant mice to restraint stress during the last week of pregnancy (2 h/day). Offspring were fed with HFD between weeks 4 and 28 of age. Prenatally stressed (PS) females and males fed with HFD showed higher body weight (females: p<0.001, n= 8; males: p<0.01, n= 8) and adipose tissue content (adipose tissue weight/body weight, both sexes: p<0.001, n= 8). Females were hyperinsulinemic (p<0.001, n= 5), with decreased expression of Foxo1 (Forkhead box protein O1) a transcription factor that plays important roles in regulation of gluconeogenesis and glycogenolysis by insulin signaling (p<0.05, n= 5) and Adiponectin (p<0.05, n= 5) in adipose tissue. On the other hand, PS males (fed with standard or HFD) had hypertriglyceridemia (p<0.001, n= 8) and hypercholesterolemia (p<0.001, n= 8). PS per se, in males, decreased the expression of Adiponectin (p<0.01, n= 5). PS animals showed a great susceptibility to develop obesity. We conclude that PS may give rise to some adverse effects, and abnormal phenotype may be provoked by or exacerbated in a later life nutritional challenge. We intend to continue our research by evaluating whether epigenetic alterations are responsible for the observed gene expression alterations

Association between diencephalic thyroliberin and arterial blood pressure in agouti-yellow and ob/ob mice may be mediated by leptin

Leptin, a hormone secreted by the adipose tissue, stimulates anorexigenic peptides and also inhibits orexigenic peptides in hypothalamic arcuate nuclei-located neurons. It also counteracts the starvation-induced suppression of thyroid hormones by up-regulating the expression of preproTRH gene. On the other hand, in addition to its role as a modulator of the thyroid-hypothalamic-hypophysial axis, thyrotropin releasing hormone (TRH) acts as a modulator of the cardiovascular system. In fact, we reported that overexpression of diencephalic TRH (dTRH) induces hypertension. We have recently shown that, in rats with obesity-induced hypertension, hyperleptinemia may produce an increase of dTRH together with an elevation of arterial blood pressure (ABP) through an increase of sympathetic activity and that these alterations were reversed by antisense oligonucleotide and small interfering RNA against preproTRH treatments. Here we explore the possible role of dTRH as a mediator involved in leptin-induced hypertension in 2 obesity mouse models: agouti-yellow mice, which are hyperleptinemic and hypertensive, and ob/ob mice, which lack functional circulating leptin. These 2 models share some characteristics, but ob/ob mice show lower ABP and plasma catecholamines levels. Then, for the first time, we report that there is a clear association between ABP and dTRH levels in both mouse models, as we have found that dTRH content was elevated in agouti-yellow mice and diminished in ob/ob mice compared with their controls. We also show that, after 3 days of subcutaneous leptin injections (10 ìg/12 hours), ABP and dTRH increased significantly in ob/ob mice with no alterations of thyroid hormone levels. These results add evidence to the putative molecular mechanisms for the strong association between obesity and hypertension.Fil: Burgueño, Adriana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Landa, Maria Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Schuman, Mariano Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Alvarez, Azucena L.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Carabelli, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Garcia, Silvia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Pirola, Carlos José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentin

Statistical analysis of the correlation between methylation levels at the promoter of candidate genes and bile acid profiling.

<p>Correlation between the status of promoter DNA methylation at CpG dinucleotides and bile acid profiling was tested using Spearman rank order correlation test.</p

Bile acid profile in healthy pregnant women and patients with intrahepatic cholestasis of pregnancy (ICP).

<p>Results are expressed as mean ± SD, P-level indicates statistical significance using Mann-Whitney test.</p><p>CA (cholic acid), CDCA (chenodeoxycholic acid), DCA (deoxycholic acid), UDCA (<b>ursodeoxycholic acid</b>), GCA (glycocholic acid), GCDCA (glycochenodeoxycholic acid), GDCA (glycodeoxycholic acid), GLCA (glycolithocholic acid), GUCDA (glycoursoeodeoxycholic acid), TCA (taurocholic acid), TCDCA (taurochenodeoxycholic acid), TDCA (taurodeoxycholic acid), TLA (taurolithocholic acid), and TUDCA (tauroursodeoxycholic acid). Bile acids species are expressed as µM.</p

In vitro exploration of the effects of the progesterone metabolite, epiallopregnanolone sulfate PM5S, on the status of CpG methylation of the <i>FXR</i>/<i>NR1H4</i> gene promoter.

<p>Huh7 human hepatoma cells were treated with an agent that inhibits DNA methylation (5 µM 5-Aza-2′-deoxicitidine), a sulphated progesterone metabolite, a reported FXR inhibitor (PM5S, 5α-Pregnan-3β-OL-20-one sulfate, sodium salt, also known as epiallopregnanolone sulfate), and a combination of both agents. Controls cells did not receive any treatment. At the top of the figure, we show data about methylation changes at the proximal and distal <i>FXR</i>/<i>NR1H4</i> gene promoter, at the bottom we show changes in the <i>FXR</i>/<i>NR1H4</i> mRNA levels.</p

Clinical, biochemical, obstetrical, and perinatal characteristics of patients with intrahepatic cholestasis of pregnancy (ICP) and healthy pregnant women.

<p>Results are expressed as mean ± SD. P-level indicates statistical significance using Mann-Whitney test.</p