Differential expression of the Wnt/β-catenin pathway in the genital tubercle (GT) of fetal male rat following maternal exposure to di-n-butyl phthalate (DBP)

Di-n-butyl phthalate (DBP) is one of the most abundantly produced endocrine disruptors that leaches out from polyvinyl chloride plastics and can cause hypospadias in male rats during maternal exposure. The objective of this study was to first explore the roles of Wnt/β-catenin pathway in the fetal rat genital tubercle (GT) following in-utero exposure to DBP. Timed-pregnant rats were given DBP by gastric intubation at a dose of 750?mg/kg body weight (bw)/day from gestation day (GD) 14 to GD18 to establish a rat model of hypospadias. On GD19, genital tubercle down-regulation of β-catenin, Phospho-GSK-3β, and up-regulation of GSK-3β (glycogen synthase kinase-3β), NFκB in fetal male rats was observed by western blot analysis. β-catenin was located in the urethral plate epithelium (UPE). Immunochemistry showed that the relative expression of β-catenin decreased in the DBP-treated fetal rat GT compared to the normal control. These findings, for the first time, indicate that DBP may affect the development of GT by down-regulating the Wnt/β-catenin pathway in fetal male rats.     

Differential expression of the Wnt/β-catenin pathway in the genital tubercle (GT) of fetal male rat following maternal exposure to di-n-butyl phthalate (DBP)

Di-n-butyl phthalate (DBP) is one of the most abundantly produced endocrine disruptors that leaches out from polyvinyl chloride plastics and can cause hypospadias in male rats during maternal exposure. The objective of this study was to first explore the roles of Wnt/β-catenin pathway in the fetal rat genital tubercle (GT) following in-utero exposure to DBP. Timed-pregnant rats were given DBP by gastric intubation at a dose of 750?mg/kg body weight (bw)/day from gestation day (GD) 14 to GD18 to establish a rat model of hypospadias. On GD19, genital tubercle down-regulation of β-catenin, Phospho-GSK-3β, and up-regulation of GSK-3β (glycogen synthase kinase-3β), NFκB in fetal male rats was observed by western blot analysis. β-catenin was located in the urethral plate epithelium (UPE). Immunochemistry showed that the relative expression of β-catenin decreased in the DBP-treated fetal rat GT compared to the normal control. These findings, for the first time, indicate that DBP may affect the development of GT by down-regulating the Wnt/β-catenin pathway in fetal male rats.     

The phytoestrogen genistein affects inflammatory-related genes expression depending on the ERα/ERβ ratio in breast cancer cells

Abstract

Breast cancer is the most common malignancy in women of developed countries. The aim of this study was to investigate the effects of the phytoestrogen genistein on the inflammatory profile in three breast cancer cell lines with different oestrogen receptors alpha (ERα) and beta (ERβ) ratio. MCF-7 (high ERα/ERβ ratio), T47D (low ERα/ERβ ratio), and MDA-MB-231 (ERα-negative) cells were treated with 1?µM of genistein for 48?h (cell proliferation and ROS production) or 4?h (mRNA expression of 18S, ERα, ERβ, pS2, Sirtuin1, IL-1β, NF-κB, COX-2, TGFβ1, PPARγ). Genistein caused a significant decrease in cell viability and an increase in ROS production in MCF-7, and the opposite happens in T47D cells. In addition, genistein rise pro-inflammatory and reduced anti-inflammatory genes expression in MCF-7, provoking the opposite effects in T47D cells. In conclusion, the phytoestrogen genistein could modulate the expression of inflammatory-related genes through its interaction with both ERs, and its effects depends on the ERα/ERβ ratio.     

Comparison of lyophilized versus liquid modified vaccinia Ankara (MVA) formulations and subcutaneous versus intradermal routes of administration in healthy vaccinia-naïve subjects

BackgroundModified vaccinia Ankara (MVA) is being developed as a safer smallpox vaccine and is being placed in the US Strategic National Stockpile (SNS) as a liquid formulation for subcutaneous (SC) administration at a dose of 1 × 10⁸ TCID₅₀ in a volume of 0.5 mL. This study compared the safety and immunogenicity of the standard formulation, dose and route with both a more stable, lyophilized formulation and with an antigen-sparing intradermal (ID) route of administration.Methods524 subjects were randomized to receive either a full dose of Lyophilized-SC, a full dose of Liquid-SC or 20% (2 × 10⁷ TCID₅₀ in 0.1 mL) of a full dose Liquid-ID MVA on Days 0 and 28. Safety and immunogenicity were followed through 180 days post second vaccination.ResultsAmong the 3 groups, the proportion of subjects with moderate/severe functional local reactions was significantly different (P = 0.0013) between the Lyophilized-SC group (30.3%), the Liquid-SC group (13.8%) and Liquid-ID group (22.0%) only after first vaccination; and for moderate/severe measured erythema and/or induration after any vaccination (P = 0.0001) between the Lyophilized-SC group (58.2%), the Liquid-SC group (58.1%) and the Liquid-ID group (94.8%) and the reactions lasted longer in the Liquid-ID group. In the ID Group, 36.1% of subjects had mild injection site skin discoloration lasting ≥6 months.After second vaccination Day (42–208), geometric mean of peak neutralization titers were 87.8, 49.5 and 59.5 for the Lyophilized-SC, Liquid-SC and Liquid-ID groups, respectively, and the maximum number of responders based on peak titer in each group was 142/145 (97.9%), 142/149 (95.3%) and 138/146 (94.5%), respectively. At 180 days after the second vaccination, geometric mean neutralization titers declined to 11.7, 10.2 and 10.4 with only 54.3%, 39.2% and 35.2% of subjects remaining seropositive for the Lyophilized-SC, Liquid-SC and Liquid-ID groups, respectively. Both the Lyophilized-SC and Liquid-ID groups were considered non-inferior (primary objective) to the Liquid-SC group.ConclusionsTransitioning to a lyophilized formulation, which has a longer shelf life, will not negatively impact immunogenicity. In a situation where insufficient vaccine is available, ID vaccination could be used, increasing the number of available doses of vaccine in the SNS 5-fold (i.e., from 20 million to 100 million doses).     

Effects of cannabinoids on neuropeptide Y and β-endorphin expression in the rat hypothalamic arcuate nucleus

The control of appetite and satiety is extremely complex and involves a balance between neurotransmitters and neuropeptides to stimulate and/or inhibit feeding behaviour. The effect of cannabinoids on food intake is well established, but little is known about the mechanism of action underlying their activity. In the present report, the effect of pharmacological manipulation of the cannabinoid receptor on the expression of hypothalamic neuropeptides is investigated. We used an immunohistochemical approach to examine the effect of intracerebroventricular administration of the cannabinoid receptor agonist WIN55,212-2 and the inverse agonist AM251 on neuropeptide Y (NPY) and the β-endorphin (β-end) neuronal hypothalamic systems. Double immunohistochemistry (c-fos/β-end) was used to assess the number of β-end neurons activated by the cannabinoid agonist. The present results showed that 1?μg WIN 55,212-2 increases β-end immunoreactivity within the arcuate nucleus while no significant changes were noted in the NPY-immunoreactive nerve fibres network in comparison to the control group. Injection of 1?μg AM251 decreases both NPY and β-end immunoreactivity within the arcuate nucleus. The number of β-end neurons exhibiting c-fos increased significantly in WIN 55,212-2 compared with the control group. These results suggest that cannabinoids affect the expression of hypothalamic neuropeptides, notably the NPY and β-end systems, which may have implications in the orexigenic action of cannabinoids.     

Vitamin a status and metabolism of benzo[α]pyrene in the rat

Male Sprague‐Dawley rats were maintained on a vitamin A‐deficient diet for a period of five weeks. At the end of that time, hepatic cytochrome P₄₅₀ levels in vitamin A‐deficient rats were 65% that of rats fed a complete diet. However, the hepatic rate of benzo[a]pyrene metabolism was significantly greater (2 times) in vitamin A‐deficient rats compared with those fed a complete diet. The pattern of metabolites separable by thin‐layer chromatography was similar in both groups of rats. Benzo[a]pyrene induced its own metabolism by a slightly greater amount in the vitamin‐sufficient rats, but it was not to the level of the deficient group, although the levels of cytochrome P₄₅₀ were still below those of the deficient rats. In discussing lung microsomes, benzo[a]pyrene pre‐treatment of deficient rats resulted in slightly elevated levels of cytochrome P₄₅₀ and a slightly greater rate of metabolism of benzo[a]pyrene compared with rats fed the complete diet.     

Feeding oxidized fat during pregnancy up-regulates expression of PPARα-responsive genes in the liver of rat fetuses

Background

Feeding oxidized fats causes activation of peroxisome proliferator-activated receptor α (PPARα) in the liver of rats. However, whether feeding oxidized fat during pregnancy also results in activation of PPARα in fetal liver is unknown. Thus, this study aimed to explore whether feeding oxidized fat during pregnancy causes a PPARα response in fetal liver. Two experiments with pregnant rats which were administered three different diets (control; oxidized fat; clofibrate as positive control) in a controlled feeding regimen during either late pregnancy (first experiment) or whole pregnancy (second experiment) were performed.

Results

In both experiments pregnant rats treated with oxidized fat or clofibrate had higher relative mRNA concentrations of the PPARα-responsive genes acyl-CoA oxidase (ACO), cytochrome P₄₅₀ 4A1 (CYP4A1), L-type carnitin-palmitoyl transferase I (L-CPT I), medium-chain acyl-CoA dehydrogenase (MCAD), and long-chain acyl-CoA dehydrogenase (LCAD) in the liver than control rats (P < 0.05). In addition, in both experiments fetuses of the oxidized fat group and the clofibrate group also had markedly higher relative mRNA concentrations of ACO, CYP4A1, CPT I, MCAD, and LCAD in the liver than those of the control group (P < 0.05), whereas the relative mRNA concentrations of PPARα, SREBP-1c, and FAS did not differ between treatment groups. In the second experiment treatment with oxidized fat also reduced triacylglycerol concentrations in the livers of pregnant rats and fetuses (P < 0.05).

Conclusion

The present study demonstrates for the first time that components of oxidized fat with PPARα activating potential are able to induce a PPARα response in the liver of fetuses. Moreover, the present study shows that feeding oxidized fat during whole pregnancy, but not during late pregnancy, lowers triacylglycerol concentrations in fetal livers.     

Alterations in liver ultrastructure and induction of UDP-glucuronyltransferase in the rat following prenatal exposure to 3,4,3′,4′-tetrachlorobiphenyl

Prenatal exposure of rats to 3,4,3,4-tetrachlorobiphenyl (4CB) at 3 mg/kg/day from day 6 through day 18 of gestation produced a high incidence of perinatal mortality. Although the fine structure of fetal liver at day 19 appeared normal, significant changes were manifest in newborn pups. These alterations included a distention of cisternae of the rough endoplasmic reticulum and conspicuous proliferation of smooth membrane elements, which persisted in survivors for several weeks. In addition, mitochondria often appeared condensed with an atypical distribution of cristae. These changes were accompanied by a postnatal induction in the activity of liver UDP-glucuronyltransferase. In treated litters, 3-week-old pups had activity levels of the enzyme three times that of the controls.     

Decreased UCP2 mRNA expression in rat stomach following vagotomy: novel role for UCP2 as free radical scavenger in the stomach?

Uncoupling protein 2 (UCP2) is a protein, located in the inner mitochondrial membrane, which dissipates the proton gradient of this membrane and uncouples respiration from oxidative phosphorylation. We found, by in situ hybridisation, UCP2 mRNA to be located in the proliferating zone of the mucous neck cells in the fundus part of the rat stomach. We also found that UCP2 expression in fundus was significantly decreased after seven days of vagotomy. Furthermore, we found manganese-containing superoxide dismutase (SOD2), in fundus, to be down-regulated in a way similar to UCP2. The amount of ATP was significantly decreased following vagotomy. It is concluded that UCP2 in the gastro-intestinal tract is regulated through vagal innervation and suggested to act as a free radical scavenger.     

Prevalence and tobacco user profile in adult population in the South of Brazil (Joaçaba, SC)

The tobacco consumption has been pointed as an epidemic and the prevention to the smoking habit is indicated as high priority for the public health of the nations. This is an observational cross-sectional study comprising a representative sample of an adult population in a Southern Brazilian city aiming to know the tobacco consumption prevalence and the profile of the smoker. Smokers' prevalence was 17.3% and female subjects were 54.9% of the sample. Smoking occurred predominantly in subjects with age older than 39 years (p<0.001), with less than eight years of education (p<0.001), with moderate and severe scores of depression measured by Beck Depression Inventory (p=0.001) and low familiar income (p=0,029). The district shows high prevalence of smokers which are predominantly subjects older than 39 years, with low income and education. This study also showed an association with moderate or severe depression and the smoking habit.     

The association between changes in lifestyle behaviors and the incidence of chronic kidney disease (CKD) in middle-aged and older men

Background

This study was designed to evaluate whether changes in lifestyle behaviors are correlated with the incidence of chronic kidney disease (CKD).

Neighborhood heterogeneity in health and well-being among the elderly in India – Evidence from Study on global AGEing and adult health (SAGE)

We establish a rationale for a multilevel approach in examining health among older adults. Using data on a nationally representative sample of 6560 Indian adults aged 50 years and older, we examine the extent of contextual variation between neighborhoods, after accounting for the compositional effect of individuals’ background characteristics, across multiple dimensions of elderly health. The variance apportioned to neighborhoods in null intercept-only models varied widely across different health outcomes examined in the elderly – while neighborhoods accounted for only 4% of the total variation in high blood pressure at exam, 23% of the total variation in self-rated poor quality of life could be attributed to neighborhood-level differences. In models that accounted for state, place of residence, and demographic and socioeconomic characteristics of individuals, the contribution of neighborhood to the total variation for most health outcomes was attenuated (2–11%) but persisted to exist. Our findings underscore the importance of neighborhoods in studying the health and well-being of the elderly in India.     

The effects of exercise training on γ-butyrobetaine hydroxylase and novel organic cation transporter-2 gene expression in the rat

The concentration of carnitine in plasma is generally increased with exercise training, suggesting that either carnitine biosynthesis is stimulated or renal reabsorption of carnitine is enhanced, or both. Carnitine, an essential cofactor in the oxidation of fatty acids, is released into the plasma following hydroxylation by γ-butyrobetaine hydroxylase (BBH), the final enzyme in the biosynthetic pathway found primarily in the liver. The organic cation transporter (OCTN2), the carnitine transporter found in kidney, is important in the distribution of carnitine by facilitating its renal reabsorption from urine. In this study, we tested the hypothesis that exercise training increases gene and protein expression of BBH and OCTN2, resulting in enhanced plasma carnitine levels. Male Wistar rats were subjected to 2 daily exercise sessions of treadmill running, 5?days per week, for a 10-week period. The concentration of total carnitine in plasma was significantly increased in trained rats compared with sedentary rats. In trained rats, mRNA and protein expression of BBH were increased in liver, whereas only BBH mRNA expression was increased in kidney. Liver of trained rats demonstrated increased mRNA and protein expression of OCTN2 compared with sedentary rats. In kidney of trained rats, however, only an increase in mRNA expression of OCTN2 was observed. Our results suggest that the improved plasma carnitine status in the trained rat is associated with increased carnitine biosynthesis in liver and kidney. The observation that OCTN2 expression was increased in kidney suggests a potential role of the kidney in the reabsorption of carnitine from the urine.     

Natural dietary ingredients (oats and alfalfa) induce covalent DNA modifications (I‐compounds) in rat liver and kidney

Mammalian tissue DNA has recently been found, via ³²P postlabeling, to contain complex profiles of age‐dependent bulky carcinogen adductlike covalent modifications, which have been termed I‐compounds, referring to their apparent indigenous origin without exposure to exogenous carcinogens. I‐compound patterns are highly species, sex, tissue, and diet specific. As shown here, the presence of certain plant ingredients in diet, i.e., ground oats and alfalfa meal, significantly contributed to the formation of these DNA derivatives.

Six groups of weanling female Sprague‐Dawley rats were fed one of the following diets for three months: a natural ingredient diet containing neither oats nor alfalfa (Wayne MRH 22/5 Rodent Blox), Wayne diet supplemented with oats or alfalfa or both, a purified semisynthetic diet (AIN‐76A), and AIN diet supplemented with oats. The natural ingredient diet produced more complex patterns and higher levels of I‐compounds than purified diet in both liver and kidney DNA. Supplementation of either diet with oats elicited the formation of four additional oats‐specific I‐compounds in liver DNA. Oats and alfalfa, individually and in combination, tended to significantly raise nonpolar and diminish polar I‐compound levels.

To determine whether the oats‐related extra spots were derived from mycotoxin contamination, two groups of rats were fed either Wayne diet or Wayne diet containing zearalenone (0.05 mg/kg) for three weeks. Zearalenone significantly increased the uterine weight but did not induce any DNA adduct formation. These findings establish a novel link between specific natural food ingredients and carcinogen adductlike DNA derivatives and may provide a model to investigate events occurring at the juncture of nutrition, metabolism, gene expression, aging, and cancer.