Viability of the use of thin-film A-SiC:H photodiodes for protein identification

In this paper, we present a multilayer device based on a-Si:H/a-SiC:H that operates as photodetector and optical filter. The use of such device in protein detection applications is relevant in Fluorescence Resonance Energy Transfer (FRET) measurements. This method demands the detection of fluorescent signals located at specific wavelengths bands in the visible part of the electromagnetic spectrum. The device operates in the visible range with a selective sensitivity dependent on electrical and optical bias. Several nanosensors were tested with a commercial spectrophotometer to assess the performance of FRET signals using glucose solutions of different concentrations. The proposed device was used to demonstrate the possibility of FRET signals detection, using visible signals of similar wavelength and intensity. The device sensitivity was tuned to enhance the wavelength band of interest using steady state optical bias at 400 nm. Results show the ability of the device to detect signals in this range. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Consumption of ultra-processed foods and their impact on the diet of young adults

OBJECTIVE To evaluate the consumption of ultra-processed foods, its associated factors, and its influence on nutrient intake in young adults.METHODS In 2004-2005, the individuals belonging to the Pelotas birth cohort of 1982 were identified for a home interview. A total of 4,297 individuals were interviewed and 4,202 individuals were included in the study (follow-up rate of 77.4%). Diet was assessed using a questionnaire on dietary intake and the percentage of daily caloric intake attributed to ultra-processed foods as well as the intake of macro- and micronutrients were estimated. The association between cohort characteristics and the consumption of ultra-processed foods was assessed using linear regression. Analysis of variance and Pearson’s Chi-square test were used to evaluate the association between the quintiles of the consumption of ultra-processed food, nutrient intake and adequacy of nutrient intake, respectively.RESULTS The consumption of ultra-processed foods corresponded to 51.2% of the total caloric intake. The consumption of ultra-processed foods was higher among women, individuals with higher education, and individuals who were never poor and eutrophic. The increased consumption of ultra-processed foods was positively correlated with the consumption of fat, cholesterol, sodium, iron, calcium, and calories (p < 0.001) and was negatively correlated with the consumption of carbohydrates, protein, and dietary fiber (p < 0.001).CONCLUSIONS The high consumption of ultra-processed foods and its positive correlation with the intake of sodium, cholesterol, and fats underscores the need to perform interventions aimed at decreasing the intake of this food group.

Insights into the mechanims underlyng the antiproliferative potential of a Co(II) coordination compound bearing 1,10-Phenanthroline-5,6-Dione: DNA and protein interaction studies

The very high antiproliferative activity of [Co(Cl)(H2O)(phendione)(2)][BF4] (phendione is 1,10-phenanthroline-5,6-dione) against three human tumor cell lines (half-maximal inhibitory concentration below 1 mu M) and its slight selectivity for the colorectal tumor cell line compared with healthy human fibroblasts led us to explore the mechanisms of action underlying this promising antitumor potential. As previously shown by our group, this complex induces cell cycle arrest in S phase and subsequent cell death by apoptosis and it also reduces the expression of proteins typically upregulated in tumors. In the present work, we demonstrate that [Co(Cl)(phendione)(2)(H2O)][BF4] (1) does not reduce the viability of nontumorigenic breast epithelial cells by more than 85 % at 1 mu M, (2) promotes the upregulation of proapoptotic Bax and cell-cycle-related p21, and (3) induces release of lactate dehydrogenase, which is partially reversed by ursodeoxycholic acid. DNA interaction studies were performed to uncover the genotoxicity of the complex and demonstrate that even though it displays K (b) (+/- A standard error of the mean) of (3.48 +/- A 0.03) x 10(5) M-1 and is able to produce double-strand breaks in a concentration-dependent manner, it does not exert any clastogenic effect ex vivo, ruling out DNA as a major cellular target for the complex. Steady-state and time-resolved fluorescence spectroscopy studies are indicative of a strong and specific interaction of the complex with human serum albumin, involving one binding site, at a distance of approximately 1.5 nm for the Trp214 indole side chain with log K (b) similar to 4.7, thus suggesting that this complex can be efficiently transported by albumin in the blood plasma.

Dietary patterns in pregnancy and birth weight

OBJECTIVE To analyze if dietary patterns during the third gestational trimester are associated with birth weight.METHODS Longitudinal study conducted in the cities of Petropolis and Queimados, Rio de Janeiro (RJ), Southeastern Brazil, between 2007 and 2008. We analyzed data from the first and second follow-up wave of a prospective cohort. Food consumption of 1,298 pregnant women was assessed using a semi-quantitative questionnaire about food frequency. Dietary patterns were obtained by exploratory factor analysis, using the Varimax rotation method. We also applied the multivariate linear regression model to estimate the association between food consumption patterns and birth weight.RESULTS Four patterns of consumption – which explain 36.4% of the variability – were identified and divided as follows: (1) prudent pattern (milk, yogurt, cheese, fruit and fresh-fruit juice, cracker, and chicken/beef/fish/liver), which explained 14.9% of the consumption; (2) traditional pattern, consisting of beans, rice, vegetables, breads, butter/margarine and sugar, which explained 8.8% of the variation in consumption; (3) Western pattern (potato/cassava/yams, macaroni, flour/farofa/grits, pizza/hamburger/deep fried pastries, soft drinks/cool drinks and pork/sausages/egg), which accounts for 6.9% of the variance; and (4) snack pattern (sandwich cookie, salty snacks, chocolate, and chocolate drink mix), which explains 5.7% of the consumption variability. The snack dietary pattern was positively associated with birth weight (β = 56.64; p = 0.04) in pregnant adolescents.CONCLUSIONS For pregnant adolescents, the greater the adherence to snack pattern during pregnancy, the greater the baby’s birth weight.

Factors associated with dietary supplement use by people who exercise at gyms

OBJECTIVE To assess the factors associated with the use of dietary supplements by people who exercise at gyms.METHODS A cross-sectional study with a sample defined by convenience, considering the number of gyms registered in the Conselho Regional de Educação Física (Regional Council of Physical Education) of Sao Luis, MA, Northeastern Brazil, from July 2011 to July 2012. The final sample comprised 723 individuals who exercise at gyms. The dependent variable was supplement use, and the explanatory variables were length of time and motivation of the physical exercises, duration, goal and self-perception of training, weekly frequency of gym attendance, sex, age, educational attainment, self-perception of body weight, smoking and self-perception of diet. The association between variables was analysed by hierarchical Poisson regression based on a theoretical model.RESULTS Supplement use was reported by 64.7% of the participants. Most of the sample was male (52.6%). The most frequent age group was 20 to 39 years (74.4%). Most participants (46.1%) had been exercising for over a year. The following variables were associated with supplement use: self-perceiving body weight as below ideal (p < 0.001), smoking (p < 0.001), exercising for 7 to 12 months (p = 0.028) or more than one year (p < 0.001), spending more than two hours at the gym (p = 0.051), and perceiving training as moderate (p = 0.024) or intense (p = 0.001).CONCLUSIONS The use of supplements lacks proper professional guidance, being motivated by individuals unsatisfied with their low body weight and who perceive their workout as intense, which raises the need for monitoring this population.

Localization studies of the FemXAB protein family in Staphylococcus aureus

Dissertação apresentada para a obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Protein-Polysaccharides of trametes versocolor: production and biological activities

Extracellular-(E-PPS) and intracellular-protein-polysaccharides (I-PPS) complexes were produced by Trametes versicolor in submerged cultures with different carbon sources. The highest extracellular-(EPS) and intracellular-polysaccharide (IPS) concentration in the complexes was obtained with tomato pomace culture. DPPH radical scavenging for E-PPS and I-PPS produced by liter of culture was equivallent to 2.115 +/- A 0.227 and 1.374 +/- A 0.364 g of ascorbic acid, respectively. These complexes showed a protector effect in the oxidation of erythrocyte membranes and had ability to inhibit the hemolysis and methemoglobin synthesis in stressed erythrocytes. These results suggest that extracellular- and intracellular- polysaccharides produced are important bioactive compounds with medicinal potential.

Production of recombinant protein hLIF in static and dynamic systems of animal cell culture

Thesis for the Degree of Master of Science in Biotechnology Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Integrating protein structural information

Dissertação apresentada para obtenção de Grau de Doutor em Bioquímica,Bioquímica Estrutural, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Protein folding, metal ions and conformational states: the case of a di-cluster ferredoxin

Dissertation presented to obtain the PhD degree in Biochemistry at the Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

Phenylketonuria: Protein content and amino acids profile of dishes for phenylketonuric patients. The relevance of phenylalanine

Phenylketonuria is an inborn error of metabolism, involving, in most cases, a deficient activity of phenylalanine hydroxylase. Neonatal diagnosis and a prompt special diet (low phenylalanine and natural-protein restricted diets) are essential to the treatment. The lack of data concerning phenylalanine contents of processed foodstuffs is an additional limitation for an already very restrictive diet. Our goals were to quantify protein (Kjeldahl method) and amino acid (18) content (HPLC/fluorescence) in 16 dishes specifically conceived for phenylketonuric patients, and compare the most relevant results with those of several international food composition databases. As might be expected, all the meals contained low protein levels (0.67–3.15 g/100 g) with the highest ones occurring in boiled rice and potatoes. These foods also contained the highest amounts of phenylalanine (158.51 and 62.65 mg/100 g, respectively). In contrast to the other amino acids, it was possible to predict phenylalanine content based on protein alone. Slight deviations were observed when comparing results with the different food composition databases.

Nutritional composition of low protein and phenylalanine-restricted dishes prepared for phenylketonuric patients

Nutritional management is essential for Phenylketonuria (PKU) treatment, consisting in a semi-synthetic and low phenylalanine (Phe) diet, which includes strictly controlled amounts of low protein natural foods (essentially fruits and vegetables) supplemented with Phe-free protein substitutes and dietetic low-protein products. PKU diet has to be carefully planned, providing the best ingredient combinations, so that patients can achieve good metabolic control and an adequate nutritional status. Hereupon, it is mandatory to know the detailed composition of natural and/or cooked foodstuffs prepared specifically for these patients. We intended to evaluate sixteen dishes specifically prepared for PKU patients, regarding the nutritional composition, Phe and tyrosine (Tyr) contents, fatty acids profile, and vitamins E and B12 amounts. The nutritional composition of the cooked samples was 15.5–92.0 g/100 g, for moisture; 0.7–3.2 g/100 g, for protein; 0.1–25.0 g/100 g, for total fat; and 5.0–62.0 g/100 g, for total carbohydrates. Fatty acids profile and vitamin E amount reflected the type of fat used. All samples were poor in vitamin B12 (0.3–0.8 μg/100 g). Boiled rice presented the highest Phe content: 50.3 mg/g of protein. These data allow a more accurate calculation of the diet portions to be ingested by the patients according to their individual tolerance.

Non-enzymatic assay for glucose by using immobilized whole-cells of E. coli containing glucose binding protein fused to fluorescent proteins

Glucose monitoring in vivo is a crucial issue for gaining new understanding of diabetes. Glucose binding protein (GBP) fused to two fluorescent indicator proteins (FLIP) was used in the present study such as FLIP-glu- 3.2 mM. Recombinant Escherichia coli whole-cells containing genetically encoded nanosensors as well as cell-free extracts were immobilized either on inner epidermis of onion bulb scale or on 96-well microtiter plates in the presence of glutaraldehyde. Glucose monitoring was carried out by Förster Resonance Energy Transfer (FRET) analysis due the cyano and yellow fluorescent proteins (ECFP and EYFP) immobilized in both these supports. The recovery of these immobilized FLIP nanosensors compared with the free whole-cells and cell-free extract was in the range of 50–90%. Moreover, the data revealed that these FLIP nanosensors can be immobilized in such solid supports with retention of their biological activity. Glucose assay was devised by FRET analysis by using these nanosensors in real samples which detected glucose in the linear range of 0–24 mM with a limit of detection of 0.11 mM glucose. On the other hand, storage and operational stability studies revealed that they are very stable and can be re-used several times (i.e. at least 20 times) without any significant loss of FRET signal. To author's knowledge, this is the first report on the use of such immobilization supports for whole-cells and cell-free extract containing FLIP nanosensor for glucose assay. On the other hand, this is a novel and cheap high throughput method for glucose assay.

Schistosoma mansoni circulating polysaccharide and protein antigens recognized by sheep antisera in patients with different clinical forms of schistosomiasis before and after treatment

Two sheep antisera, one of which raised against polysaccharide (Po) and other against protein (Pt) components of Schistosoma mansoni adult worms, were assessed by ELISA for their ability to detect circulating parasite antigens in patients with different clinical forms of chronic schistosomiasis mansoni. The former antiserum detected parasite antigens in liver granulomata and the latter in renal glomeruli from schistosomiasis patients and mice experimentally infected with S. mansoni. In general, the levels and/or positivity rate of circulating antigens and specific IgG antibodies were significantly higher in patients with hepatointestinal (HI) and hepatosplenic (HS) forms than in mild intestinal (I) forms. An association between Po antigens and clinical features of the disease was observed, as the level of these antigens was low (137 ng/ml) as well as the positivity rate (7.9%) in patients with I forms; values that were intermediate (593 ng/ml and 33.3%) in those with HI forms, and high (1.563 ng/ml and 50.0%) in more severe HS forms. The Pt antigens were detected in the studied clinical forms not differing statistically but, the positivity rate was significantly higher in HS forms comparatively to I forms. The antisera studied revealed distinct circulating antigen profiles, and the prognostic value of Po and Pt antigens was suggested.