Mouse Testicular Mkrn3 Expression Is Primarily Interstitial, Increases Peripubertally, and Is Responsive to LH/hCG.

Studies in humans and mice support a role for Makorin RING finger protein 3 (MKRN3) as an inhibitor of gonadotropin-releasing hormone (GnRH) secretion prepubertally, and its loss of function is the most common genetic cause of central precocious puberty in humans. Studies have shown that the gonads can synthesize neuropeptides and express MKRN3/Mkrn3 mRNA. Therefore, we aimed to investigate the spatiotemporal expression pattern of Mkrn3 in gonads during sexual development, and its potential regulation in the functional testicular compartments by gonadotropins. Mkrn3 mRNA was detected in testes and ovaries of wild-type mice at all ages evaluated, with a sexually dimorphic expression pattern between male and female gonads. Mkrn3 expression was highest peripubertally in the testes, whereas it was lower peripubertally than prepubertally in the ovaries. Mkrn3 is expressed primarily in the interstitial compartment of the testes but was also detected at low levels in the seminiferous tubules. In vitro studies demonstrated that Mkrn3 mRNA levels increased in human chorionic gonadotropin (hCG)-treated Leydig cell primary cultures. Acute administration of a GnRH agonist in adult mice increased Mkrn3 expression in testes, whereas inhibition of the hypothalamic-pituitary-gonadal axis by chronic administration of GnRH agonist had the opposite effect. Finally, we found that hCG increased Mkrn3 mRNA levels in a dose-dependent manner. Taken together, our developmental expression analyses, in vitro and in vivo studies show that Mkrn3 is expressed in the testes, predominantly in the interstitial compartment, and that Mkrn3 expression increases after puberty and is responsive to luteinizing hormone/hCG stimulation.

The addition of residue from pruning of yerba mate (Ilex paraguariensis) in laying hens modulates fatty acid profile and incorporates chlorogenic acid in the egg.

This study aimed to determine whether the addition of Ilex paraguariensis residue to the basal diet of laying hens improves bird health, production, fatty acids, egg quality, and transfer of chlorogenic acids to the egg. One hundred twenty birds of the Hy-line lineage were used, divided into five groups with six replicates and four birds per replicate, as follows: T0 (control group, without IPPR supplementation), T0.5 (basal diet plus 0.5% of IPPR), T1 (basal diet plus 1% IPPR), T1.5 (basal diet plus 1.5% IPPR), and T2 (basal diet plus 2% IPPR), for two consecutive cycles of 21 days each. The productive performance of the birds, physicochemical composition of eggs, fatty acid profile, the concentration of chlorogenic acids in eggs, leukocyte count, serum biochemistry, oxidant and antioxidant status, total bacterial count in eggs, and feces were evaluated at 21 and 42 days of the experiment. The addition of IPPR to the birds' diet no-change eggs production and feed conversion. In the egg yolks of birds that consumed IPPR, there were chlorogenic acids. There was a reduction in the total bacterial count in the feces and eggshells of the birds that received 2% of the IPPR. IPPR intake also reflected a reduction in total saturated fatty acid levels and increased monounsaturated fatty acids. These findings suggest that IPPR is a compelling alternative for laying hens; however, its ingestion also has negative effects that are discussed in below. Chlorogenic acids in the egg are desirable, as these phenolics have nutraceutical effects in humans.

Dietary supplementation with nerolidol improves the antioxidant capacity and muscle fatty acid profile of Brycon amazonicus exposed to acute heat stress.

An increase in water temperature in the Amazon River has elicited concerns about commercially important fish species associated with food security, such as matrinxã (Brycon amazonicus). Studies have demonstrated the positive effects of diets supplemented with plant-based products that combat heat stress-induced oxidative damage. The aim of this study was to determine whether dietary supplementation with nerolidol prevents or reduces muscle oxidative damage and impairment of the fillet fatty acid profile of matrinxã exposed to heat stress. Plasma and muscle reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while plasma superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity was significantly lower. The total content of saturated fatty acids (SFA) in fillets was significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while he total content of polyunsaturated fatty acids (PUFA) was significantly lower. Nerolidol prevented the increase of muscle LPO and plasma ROS and LPO levels in fish exposed to heat stress, and partially prevented the increase in muscle ROS levels. Diets containing nerolidol prevented the inhibition of muscle GPx activity in fish exposed to heat stress, and partially prevented the decrease of plasma GPx activity. The nerolidol-supplemented diet prevented the increase of fillet SFA in fish exposed to heat stress, while partially preventing the decrease of PUFA. We conclude that acute heat stress at 34 °C for 72 h causes plasma and muscular oxidative damage, and that homeoviscous adaptation to maintain membrane fluidity can represent a negative impact for fish consumers. A nerolidol diet can be considered a strategy to prevent heat stress-induced oxidative damage and impairment of muscle fatty acid profiles.

Inclusion of a phytogenic bend in broiler diet as a performance enhancer and anti-aflatoxin agent: Impacts on health, performance, and meat quality.

The objective of this study was to determine whether a phytogenic blend (PB), formulated based on organic acids, tannins, curcumin, and essential oils, could replace the antimicrobials commonly used as growth promoters in the poultry industry without compromising zootechnical performance, health, or meat quality. In addition, our goal was to report the anti-aflatoxin effect of this phytogenic blend. Four treatments were used: TC, or control; T250, T500, and T1000, representing test doses of 250, 500, 1000 mg PB/kg of feed, respectively, or a 34-day experiment (initial and growth phases). On day 22 of the study and age of the birds, 500 ppb of aflatoxin was included in the diet to represent an intestinal challenge and to evaluate the growth-promoting effects of PB. In the initial phase (up to 21 days), there were no differences between groups in weight gain, feed intake, or feed conversion. After adding an aflatoxin-contaminated feed, doses of 250 and 500 mg/kg minimized the adverse effects on feed consumption and feed conversion caused by aflatoxin; but 1000 mg/kg did not differ between groups. In birds that consumed PB (T250, T500, and T1000) compared to the control, there were the following changes: 1) lower counts of heterophiles, lymphocytes, and monocytes; 2) lower lipid peroxidation and high non-protein thiols levels in breast meat; 3) lower bacteria counts in broiler litter; and 4) lower ALT levels. Greater intestinal villus/crypt ratios were observed at T250 and T500. The dose of 250 mg/kg reduced saturated fatty acids and increased unsaturated fatty acids. The chemical-physical composition of the meat did not differ between treatments. The findings suggest that the addition of a PB has a high potential to improve performance for chickens in the growing stage and minimize the adverse effects of aflatoxicosis.

ERK1/2 phosphorylation and IL-6 production are involved in the differential susceptibility to Toxoplasma gondii infection in three types of human (cyto/ syncytio/ extravillous) trophoblast cells.

During pregnancy, Toxoplasma gondii can triggers serious manifestations and potentially affect the fetal development. In this scenario, differences in susceptibility of trophoblast cells to T. gondii infection might be evaluated in order to establish new therapeutic approaches capable of interfering in the control of fetal infection by T. gondii. This study aimed to evaluate the susceptibility of cytotrophoblast, syncytiotrophoblast and extravillous trophoblast cells to T. gondii infection. Our data demonstrate that HTR-8/SVneo cells (extravillous trophoblast cells) present higher susceptibility to T. gondii infection when compared to syncytiotrophoblast and cytotrophoblast cells, whereas syncytiotrophoblast was the cell type more resistant to the parasite infection. Also, cytotrophoblast and syncytiotrophoblast cells produced significantly more IL-6 than HTR-8/SVneo cells. On the other hand, HTR-8/SVneo cells showed higher ERK1/2 phosphorylation than cytotrophoblast and syncytiotrophoblast cells. ERK1/2 inhibition reduced T. gondii infection and increased IL-6 production in HTR-8/SVneo cells. Thus, it is plausible to conclude that the greater susceptibility of HTR-8/SVneo cells to infection by T. gondii is related to a higher ERK1/2 phosphorylation and lower levels of IL-6 in these cells compared to other cells, suggesting that these mediators may be important to favor the parasite infection in this type of trophoblastic population.

Effects of biocholine powder supplementation in ewe lambs: Growth, rumen fermentation, antioxidant status, and metabolism.

The objective of this study was to evaluate the effects of biocholine powder (BP) added to the concentrate of ewe lambs on growth, rumen fermentation, antioxidant status, and metabolism. Forty-eight Lacaune lambs were divided 3 treatments: T0, T4, and T8, representing 0, 4, and 8 g of BP per animal/day added to the concentrate, respectively. The T4 and T8 lambs had greater average daily gain from d 0-75 (0.22, 0.25, and 0.26 kg/d for T0, T4 and T8, respectively). T4 and T8 lambs had higher methylene blue test results (indicating microbiological activity), and the higher effects were detected for T8 lambs. The BP in lamb's diet increased the ruminal concentration of propionate (T4 and T8). The T8 lambs had greater serum activity of superoxide dismutase and glutathione transferase, and lower reactive oxygen species levels. We conclude that BP is a biotechnological additive with high nutritional potential.

Liraglutide Activates Type 2 Deiodinase and Enhances ß3-Adrenergic-Induced Thermogenesis in Mouse Adipose Tissue.

Aims: Liraglutide is a long-acting glucagon-like peptide 1 (GLP-1) receptor agonist used as an anti-hyperglycemic agent in type 2 diabetes treatment and recently approved for obesity management. Weight loss is attributed to appetite suppression, but therapy may also increase energy expenditure. To further investigate the effect of GLP-1 signaling in thermogenic fat, we assessed adipose tissue oxygen consumption and type 2 deiodinase (D2) activity in mice treated with liraglutide, both basally and after ß3-adrenergic treatment. Methods: Male C57BL/6J mice were randomly assigned to receive liraglutide (400 µg/kg, n=12) or vehicle (n=12). After 16 days, mice in each group were co-treated with the selective ß3-adrenergic agonist CL316,243 (1 mg/kg, n=6) or vehicle (n=6) for 5 days. Adipose tissue depots were assessed for gene and protein expression, oxygen consumption, and D2 activity. Results: Liraglutide increased interscapular brown adipose tissue (iBAT) oxygen consumption and enhanced ß3-adrenergic-induced oxygen consumption in iBAT and inguinal white adipose tissue (ingWAT). These effects were accompanied by upregulation of UCP-1 protein levels in iBAT and ingWAT. Notably, liraglutide increased D2 activity without significantly upregulating its mRNA levels in iBAT and exhibited additive effects to ß3-adrenergic stimulation in inducing D2 activity in ingWAT. Conclusions: Liraglutide exhibits additive effects to those of ß3-adrenergic stimulation in thermogenic fat and increases D2 activity in BAT, implying that it may activate this adipose tissue depot by increasing intracellular thyroid activation, adding to the currently known mechanisms of GLP-1A-induced weight loss.

Consequences of oxidative damage on the fatty acid profile in muscle of Cichlasoma amazonarum acutely exposed to copper.

Rapid industrialization results in the production of large quantities of waste that are commonly discharged into water bodies, leading to the damage of the aquatic ecosystem and freshwater organisms. Copper (Cu) can induce oxidative damage in fish muscle, the main fish portion that is consumed by humans. However, the responses of the Amazon fish Cichlasoma amazonarum and its capacity to withstand acute Cu concentrations found in Amazon water around mines remain unknown. Thus, the aim of this study was to evaluate whether exposure to Cu causes muscle oxidative stress and/or oxidative damage and impairs the fillet fatty acid profile of C. amazonarum acutely exposed to Cu found in Amazon waters around mines. Muscle reactive oxygen species and protein carbonylation levels were significantly higher in fish exposed to 1500 µg/L Cu compared with the control group, while muscle lipid peroxidation levels were significantly higher in fish exposed to 500, 750, and 1500 µg/L Cu compared with control group. Muscle antioxidant capacity against peroxyl radical's levels and glutathione peroxidase activity were significantly lower in fish exposed to 1500 µg/L Cu compared with the control group, while muscle superoxide dismutase activity was significantly lower in fish exposed to 750 and 1500 µg/L Cu compared with control group. The total content of saturated fatty acids was significantly higher in fish exposed to 1500 µg/L Cu compared with the control group, while the total content of monounsaturated fatty acids and sum of n3 fatty acids were significantly lower in fish exposed to 1500 µg/L Cu compared with control group. No significant difference was observed regarding muscle catalase, glutathione S-transferase, and glutathione reductase activities. Based on these lines of evidence, the results of this comprehensive study agree with the initial hypothesis that the exposure to Cu found in Amazon water around mines induces oxidative damage and inhibits enzymatic and non-enzymatic antioxidant response in the muscle of C. amazonarum exposed to high Cu levels. Moreover, the impairment of the fillet fatty acid profile appears to be mediated by oxidative damage, representing a negative impact on fish health.

Addition of pooled pumpkin seed to mixed meals reduced postprandial glycemia: a randomized placebo-controlled clinical trial.

We investigated if pumpkin and flaxseeds could improve postprandial glycemic, food intake, and appetitive responses. Herein, we hypothesize based on the literature that pumpkin seed has potential to lower postprandial glycemic effects. Therefore, we conducted a randomized, single-blind, placebo-controlled, crossover design study involving normoglycemic adults (food intake: n = 25; glycemia: n = 15). Three high-carbohydrate mixed meals presenting no seed (control [C]) or 65 g of the tested seeds (pumpkin seed [P] or flaxseed [F]) were consumed in 3 nonconsecutive days. Test meals had similar nutritional composition. Blood glucose was measured by capillary finger blood at 0 (immediately before), 15, 30, 45, 60, 90, and 120 minutes after the ingestion of each meal, and the incremental area under glycemic response curves (iAUC) were calculated. Appetitive responses were assessed, and dietary records were used to evaluate food intake on testing days. Glucose iAUC was significantly lower in P compared with C (reduction of ~35%, P = .025). There was no significant differences in glucose iAUC between F and C (P = .257). Glycemic response at each time point did not differ between C, P, and F (Pgroup × time = .238). Fiber consumption was higher in F (P = .009) than in C, but there were no differences in appetitive responses, energy, or macronutrient consumptions between dietary interventions. Acute consumption of 65 g of pumpkin seed markedly reduced postprandial glycemia. Pumpkin seed has potential as a hypoglycemic food, which now deserves to be confirmed in long-term studies.

GQ-16, a TZD-Derived Partial PPARγ Agonist, Induces the Expression of Thermogenesis-Related Genes in Brown Fat and Visceral White Fat and Decreases Visceral Adiposity in Obese and Hyperglycemic Mice.

BACKGROUND: Beige adipocytes comprise a unique thermogenic cell type in the white adipose tissue (WAT) of rodents and humans, and play a critical role in energy homeostasis. In this scenario, recruitment of beige cells has been an important focus of interest for the development of novel therapeutic strategies to treat obesity. PPARγ activation by full agonists (thiazolidinediones, TZDs) drives the appearance of beige cells, a process so-called browning of WAT. However, this does not translate into increased energy expenditure, and TZDs are associated with weight gain. Partial PPARγ agonists, on the other hand, do not induce weight gain, but have not been shown to drive WAT browning. The present study was designed to investigate the effects of GQ-16 on BAT and on browning of WAT in obese mice. METHODS: Male Swiss mice with obesity and hyperglycemia induced by high fat diet were treated with vehicle, rosiglitazone (4 mg/kg/d) or the TZD-derived partial PPARγ agonist GQ-16 (40 mg/kg/d) for 14 days. Fasting blood glucose, aspartate aminotransferase, alanine aminotransferase and lipid profile were measured. WAT and brown adipose tissue (BAT) depots were excised for determination of adiposity, relative expression of Ucp-1, Cidea, Prdm16, Cd40 and Tmem26 by RT-qPCR, histological analysis, and UCP-1 protein expression analysis by immunohistochemistry. Liver samples were also removed for histological analysis and determination of hepatic triglyceride content. RESULTS: GQ-16 treatment reduced high fat diet-induced weight gain in mice despite increasing energy intake. This was accompanied by reduced epididymal fat mass, reduced liver triglyceride content, morphological signs of increased BAT activity, increased expression of thermogenesis-related genes in interscapular BAT and epididymal WAT, and increased UCP-1 protein expression in interscapular BAT and in epididymal and inguinal WAT. CONCLUSION: This study suggests for the first time that a partial PPARγ agonist may increase BAT activity and induce the expression of thermogenesis-related genes in visceral WAT. GENERAL SIGNIFICANCE: These findings suggest that PPARγ activity might be modulated by partial agonists to induce WAT browning and treat obesity.

Appropriate time interval to repeat ambulatory blood pressure monitoring in patients with white-coat resistant hypertension.

Resistant hypertension is defined as uncontrolled office blood pressure, despite the use of ≥3 antihypertensive drugs. Ambulatory blood pressure monitoring (ABPM) is mandatory to diagnose 2 different groups, those with true and white-coat resistant hypertension. Patients are found to change categories between controlled/uncontrolled ambulatory pressures without changing their office blood pressures. In this way, ABPM should be periodically repeated. The aim of this study was to evaluate the most appropriate time interval to repeat ABPM to assure sustained blood pressure control in patients with white-coat resistant hypertension. This prospective study enrolled 198 patients (69% women; mean age: 68.9±9.9 years) diagnosed as white-coat resistant hypertension on ABPM. Patients were submitted to a second confirmatory examination 3 months later and repeated twice at 6-month intervals. Statistical analyses included Bland-Altman repeatability coefficients and multivariate logistic regression. Mean office blood pressure was 163±20/84±17 mm Hg, and mean 24-hour blood pressure was 118±8/66±7 mm Hg. White-coat resistant hypertension diagnosis presented a moderate reproducibility and was confirmed in 144 patients after 3 months. In the third and fourth ABPMs, 74% and 79% of patients sustained the diagnosis. In multivariate regression, a daytime systolic blood pressure ≤115 mm Hg in the confirmatory ABPM triplicated the chance of white-coat resistant hypertension status persistence after 1 year. In conclusion, a confirmatory ABPM is necessary after 3 months of the first white-coat-resistant hypertension diagnosis, and the procedure should be repeated at 6-month intervals, except in patients with daytime systolic blood pressure ≤115 mm Hg, in whom it may be repeated annually.

Interdisciplinary approach improves nutritional status of children with heart diseases.

OBJECTIVES: We evaluated the role of an interdisciplinary team in improving the nutrient intake and nutritional status of children with heart diseases. METHODS: Thirty-five children 10.0 +/- 7.5 mo old were studied over a 6-mo period with regard to anthropometric measurements and dietary intake. RESULTS: On admission, malnutrition prevalence was 57% and vitamin and mineral intakes were below estimated average requirement levels (dietary reference intakes) in the majority of patients. Stunting and wasting were more frequent among patients with pulmonary hypertension and/or uncompensated congestive heart failure. Follow-up analyses showed increased Z scores of weight for height (-1.17 +/- 1.03 versus -0.32 +/- 1.08, P < 0.01) and height for age (-1.09 +/- 0.96 versus -0.51 +/- 1.36, P < 0.01). In patients with pulmonary hypertension or uncompensated congestive heart failure, the height-for-age index remained unchanged. Energy intake did not differ (112.0 +/- 20.4 and 119.0 +/- 18.0 kcal.kg(-1).d(-1)) and significant increases were seen in intakes of micronutrients and minerals over the study period. CONCLUSION: Intervention by an interdisciplinary team improved the nutrient intake and nutritional status of patients overall but was insufficient to improve growth in the subgroup with pulmonary hypertension or uncompensated congestive heart failure. Nutritional support should be made routine in the treatment of children with heart disease.

Prevalência de asma em escolares de 13-14 anos de Brasília - DF. Comparação entre 1996 e 2002, por grupos socioeconômicos / Asthma prevalence in 13-14-year-old schoolchildren from Brasília - DF. Comparison between 1996 and 2002 data by socioeconomic status

Objetivos: Determinar a prevalência da asma e seus sintomas em escolares de 13-14 anos de idade, de Brasília-DF. Avaliar suas tendências, comparando os dados de 2002 com os de 1996. Comparar as taxas de prevalência da asma entre diferentes grupos socioeconômicos. Métodos: Dois estudos de corte transversal foram realizados com intervalo de seis anos, usando o questionário escrito do protocolo ISAAC (Fases I e III). Nesta pesquisa, 39 escolas públicas e privadas foram escolhidas aleatoriamente, em oito regiões administrativas de Brasília, que foram divididas em três grupos, segundo as condições socioeconômicas da população. Resultados: Foram obtidos 3.009 questionários, sendo 80% de alunos de escolas públicas, com 53,5 % do sexo feminino. Comparando com dados de 1996, não houve aumento significativo da prevalência de asma diagnosticada (de 13,8 para 14,8%) e nem de asma atual (de 19,5 para 19,7%). A prevalência de asma diagnosticada foi significativamente maior nas escolas privadas do que nas escolas públicas (17,8% x 14,1 %), assim como nas populações de melhor nível socioeconômico (17,7% x 11,5%). Comparando os grupos socioeconômicos, não houve diferença significativa entre os dois estudos. Houve relato concomitante de rinite alérgica em 37,4% dos asmáticos e de eczema atópico em 24,7%. Apenas 53 (1,7%) reportaram a ocorrência das três patologias simultaneamente. Conclusão: A prevalência de asma diagnosticada, em crianças de 13 a 14 anos de idade do DF, não apresentou aumento significativo em um período de seis anos e foi significativamente maior nas crianças de escolas privadas e de nível socioeconômico mais elevado.