Rat visceral yolk sac cells: viability and expression of cell markers during maternal diabetes

The function of the visceral yolk sac (VYS) is critical for embryo organogenesis until final fetal development in rats, and can be affected by conditions such as diabetes. In view of the importance of diabetes during pregnancy for maternal and neonatal health, the objective of this study was to assess fetal weight, VYS cell markers, and viability in female Wistar rats (200-250 g) with induced diabetes (alloxan, 37 mg/kg) on the 8th gestational day (gd 8). At gd 15, rats from control (n=5) and diabetic (n=5) groups were anesthetized and laparotomized to remove the uterine horns for weighing of fetuses and collecting the VYS. Flow cytometry was used for characterizing VYS cells, and for determining mitochondrial activity, cell proliferation, DNA ploidy, cell cycle phases, and caspase-3 activity. Fetal weight was reduced in the diabetic group. Expression of the cell markers CD34, VEGFR1, CD115, CD117, CD14, CCR2, CD90, CD44, STRO-1, OCT3/4, and Nanog was detected in VYS cells in both groups. In the diabetic group, significantly decreased expression of CD34 (P<0.05), CCR2 (P<0.001), and OCT3/4 (P<0.01), and significantly increased expression of CD90 (P<0.05), CD117 (P<0.01), and CD14 (P<0.05) were observed. VYS cells with inactive mitochondria, activated caspase-3, and low proliferation were present in the rats with diabetes. Severe hyperglycemia caused by maternal diabetes had negative effects on pregnancy, VYS cell viability, and the expression of cell markers.

Chemical markers and antifungal activity of red propolis from Sergipe, Brazil

The aim of this study was to analyze the physicochemical properties and antifungal activities of the red propolis samples from Sergipe, Brazil, and also evaluate their variability throughout the year. The characterization of the hydroalcoholic extract (HPE) of the red propolis samples was performed monthly from October 2009 to September 2010. The concentrations of the bioactive compounds varied during the year, but their chromatographic profiles were similar. Four compounds were identified by comparison with authentic standards. Formononetin was one of the predominant compounds in all propolis extracts. In our study, it was observed that all the propolis samples inhibited the growth of Candida species. Multivariate analysis confirmed the variations in chemical composition and color of the HPEs throughout the year. The biological activities of the HPEs were statistically significant (p<0.05), and all samples exhibited antifungal properties.

The association of markers of oxidative-inflammatory status with malnutrition in hemodialysis patients with serum ferritin lower than 500 ng/mL

INTRODUCTION: Enhanced inflammatory-oxidative status is well established in chronic kidney disease. OBJECTIVE: The objective of this study was to evaluate the oxidative- inflammatory status and iron indices in patients undergoing maintenance hemodialysis (HD) with serum ferritin lower than 500ng/mL, and to correlate them with nutritional status. METHOD: In a cross-sectional survey 35 HD patients (23 with normal nutritional status, 12 with Protein-Energy-Wasting syndrome, PEW), and healthy volunteers (n = 35) were studied. Serum concentration of iron, ferritin, transferrin saturation, malondialdehyde (MDA), protein carbonyl (PC), high-sensitive serum C -reactive protein (hs-CRP) and blood counts were determined. The nutritional status was determined by anthropometric and biochemical criteria. RESULTS: HD patients showed low values of hemoglobin and higher values of ferritin, MDA and PC when compared with healthy volunteers. HD subjects with PEW had higher values of PC and hs-PCR as compared to HD patients with normal nutritional status. A multiple logistic regression analysis showed that the independent variables PC (Wald Statistic 4.25, p = 0.039) and hs-CRP (Wald Statistic 4.83, p = 0.028) where related with the patients' nutritional condition. CONCLUSION: In HD patients with serum ferritin below 500 ng/mL was observed one association of the markers of oxidative stress and inflammation with poor nutritional status independently of serum ferritin, gender and age.

Effects of grape powder supplementation on inflammatory and antioxidant markers in hemodialysis patients: A randomized double-blind study

Introduction: Polyphenols contained in natural sources such as grapes, have been considered pharmacological agents to combat oxidative stress and inflammation, common features in Chronic Kidney Disease patients. Objective: To evaluate the effects of grape powder supplementation on inflammatory and antioxidant biomarkers in hemodialysis (HD) patients. Methods: The double-blind placebo-controlled randomized clinical trial evaluated non-diabetic HD patients that received grape powder (500 mg of polyphenols/day) (n = 16, 9 men, 53.0 ± 9.8 years of age, 111.6 ± 58.2 HD months) or placebo (n = 16, 9 men, 52.7 ± 13.7 years of age, 110.4 ± 93.1 HD months) for five weeks. The glutathione peroxidase (GSH-Px) activity and C-reactive protein (CRP) levels were evaluated by ELISA method. Results: After the intervention period, the patients receiving grape powder showed an increase in the GSH-Px activity (16.5 (41.0) to 42.0 (43.3) nmol/min/ml) (p < 0.05) and they did not have the CRP levels increased as seen in placebo group (2.6 (0.28) to 2.8 (0.23 mg/L) (p < 0.05). Conclusion: The use of grape powder as phenolic source could play an important role as an antioxidant and anti-inflammatory agent in non-diabetic HD patients.

Genetic purity certificate in seeds of hybrid maize using molecular markers

One of the main features that confer high quality to the seed is its genetic purity, in which one of the major causes of contamination is the self-pollination of the female parent. Up to date, there is no accurate and fast methods for detecting such contamination. Thus, this work was carried out to certify the genetic purity in seeds of hybrid maize using different biochemical and DNA-based markers. Two single-cross hybrids and their parental lines derived from the maize breeding program at UFLA were evaluated by isoenzymatic pattern of alcohol dehydrogenase (ADH), esterase (EST), acid phosphatase (ACP), glutamate-oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphoglucomate dehydrogenase (PGDH), catalase (CAT) and ß-glucosidade (ßGLU) and by microsatellites markers. The enzymatic systems that were able to distinguish the hybrids from their parental line were the catalase, the isocitrate dehydrogenase and the esterase. The esterase showed a Mendelian segregation pattern for UFLA 8/3 hybrid, that enables a safer genetic purity certificate. Microsatellites were able to differentiate the hybrid lines and the respective parental lines. Moreover, this technique was fast, precise and without environment effects. For microsatellites, the amplification pattern was identical when young leaves or seeds were used as DNA source. The possibility of using seeds as DNA source would accelerate and facilitate the role process of the genetic purity analysis.

cDNA-AFLP na identificação de genes relacionados a qualidade fisiológica

Alguns trabalhos têm evidenciado a existência de genótipos de soja contrastantes para qualidade fisiológica de semente. Tais diferenças existem em virtude da presença de sementes com total ou parcial impermeabilidade do tegumento à água, o que as tornam menos susceptíveis aos danos mecânicos, as adversidades climáticas e a deterioração por umidade. A característica de tegumento semi-permeável pode ser incorporada às cultivares de alta produção por meio dos programas de melhoramento. No entanto, há necessidade de caracterizar os genes ou as regiões genômicas envolvidas com esta característica. Nesse contexto, ferramentas da biologia molecular, como a técnica de cDNA-AFLP, podem auxiliar a identificação de genes relacionados a qualidade fisiológica de sementes. O objetivo desse estudo foi verificar a eficácia da técnica de cDNA-AFLP, na obtenção de fragmentos de genes diferencialmente expressos entre tegumentos de sementes de soja com permeabilidade contrastante. Sementes provenientes dos genótipos CD-202 (tegumento amarelo, permeável e susceptível a deterioração) e TP (tegumento preto, semi-permeável e resistente a deterioração) foram cultivadas em casa-de-vegetação, sob condições homogêneas. Realizou-se a coleta das sementes em diferentes estágios de desenvolvimento (25, 30, 35, 40 e 55 dias após a antese). Procedeu-se à extração do RNA total utilizando-se três métodos, sendo o reagente Pure Link Plant RNA o mais eficiente. Para obtenção do cDNA dupla fita utilizou-se o kit Double Stranded cDNA Synthesis. A partir do cDNA obtido, aplicou-se a técnica de AFLP testando-se um total de 64 combinações de primers. Foram obtidos 47 fragmentos de cDNA diferencialmente expressos entre os tegumentos de sementes dos dois genótipos, os quais poderão ter suas funções reveladas pelo sequenciamento e análise in sílico. De acordo com os resultados obtidos, a técnica de cDNA-AFLP demonstra ser uma alternativa promissora para estudos que visem à identificação de genes relacionados a qualidade de sementes.