Leptospira interrogans activation of peripheral blood monocyte glycolipoprotein demonstrated in whole blood by the release of IL-6

Glycolipoprotein (GLP) from pathogenic serovars of Leptospira has been implicated in the pathogenesis of leptospirosis by its presence in tissues of experimental animals with leptospirosis, the inhibition of the Na,K-ATPase pump activity, and induced production of cytokines. The aims of the present study were to investigate the induction of IL-6 by GLP in peripheral blood mononuclear cells (PBMC) and to demonstrate monocyte stimulation at the cellular level in whole blood from healthy volunteers. PBMC were stimulated with increasing concentrations (5 to 2500 ng/ml) of GLP extracted from the pathogenic L. interrogans serovar Copenhageni, lipopolysaccharide (positive control) or medium (negative control), and supernatants were collected after 6, 20/24, and 48 h, and kept at -80ºC until use. Whole blood was diluted 1:1 in RPMI medium and cultivated for 6 h, with medium, GLP and lipopolysaccharide as described above. Monensin was added after the first hour of culture. Supernatant cytokine levels from PBMC were measured by ELISA and intracellular IL-6 was detected in monocytes in whole blood cultures by flow-cytometry. Monocytes were identified in whole blood on the basis of forward versus side scatter parameters and positive reactions with CD45 and CD14 antibodies. GLP ( > or = 50 ng/ml)-induced IL-6 levels in supernatants were detected after 6-h incubation, reaching a peak after 20/24 h. The percentage of monocytes staining for IL-6 increased with increasing GLP concentration. Thus, our findings show a GLP-induced cellular activation by demonstrating the ability of GLP to induce IL-6 and the occurrence of monocyte activation in whole blood at the cellular level.

L-carnitine as an ergogenic aid for patients with chronic obstructive pulmonary disease submitted to whole-body and respiratory muscle training programs

The effects of adding L-carnitine to a whole-body and respiratory training program were determined in moderate-to-severe chronic obstructive pulmonary disease (COPD) patients. Sixteen COPD patients (66 ± 7 years) were randomly assigned to L-carnitine (CG) or placebo group (PG) that received either L-carnitine or saline solution (2 g/day, orally) for 6 weeks (forced expiratory volume on first second was 38 ± 16 and 36 ± 12%, respectively). Both groups participated in three weekly 30-min treadmill and threshold inspiratory muscle training sessions, with 3 sets of 10 loaded inspirations (40%) at maximal inspiratory pressure. Nutritional status, exercise tolerance on a treadmill and six-minute walking test, blood lactate, heart rate, blood pressure, and respiratory muscle strength were determined as baseline and on day 42. Maximal capacity in the incremental exercise test was significantly improved in both groups (P < 0.05). Blood lactate, blood pressure, oxygen saturation, and heart rate at identical exercise levels were lower in CG after training (P < 0.05). Inspiratory muscle strength and walking test tolerance were significantly improved in both groups, but the gains of CG were significantly higher than those of PG (40 ± 14 vs 14 ± 5 cmH2O, and 87 ± 30 vs 34 ± 29 m, respectively; P < 0.05). Blood lactate concentration was significantly lower in CG than in PG (1.6 ± 0.7 vs 2.3 ± 0.7 mM, P < 0.05). The present data suggest that carnitine can improve exercise tolerance and inspiratory muscle strength in COPD patients, as well as reduce lactate production.

Lipopolysaccharide-induced expression of cell surface receptors and cell activation of neutrophils and monocytes in whole human blood

Lipopolysaccharide (LPS) activates neutrophils and monocytes, inducing a wide array of biological activities. LPS rough (R) and smooth (S) forms signal through Toll-like receptor 4 (TLR4), but differ in their requirement for CD14. Since the R-form LPS can interact with TLR4 independent of CD14 and the differential expression of CD14 on neutrophils and monocytes, we used the S-form LPS from Salmonella abortus equi and the R-form LPS from Salmonella minnesota mutants to evaluate LPS-induced activation of human neutrophils and monocytes in whole blood from healthy volunteers. Expression of cell surface receptors and reactive oxygen species (ROS) and nitric oxide (NO) generation were measured by flow cytometry in whole blood monocytes and neutrophils. The oxidative burst was quantified by measuring the oxidation of 2',7'-dichlorofluorescein diacetate and the NO production was quantified by measuring the oxidation of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate. A small increase of TLR4 expression by monocytes was observed after 6 h of LPS stimulation. Monocyte CD14 modulation by LPS was biphasic, with an initial 30% increase followed by a 40% decrease in expression after 6 h of incubation. Expression of CD11b was rapidly up-regulated, doubling after 5 min on monocytes, while down-regulation of CXCR2 was observed on neutrophils, reaching a 50% reduction after 6 h. LPS induced low production of ROS and NO. This study shows a complex LPS-induced cell surface receptor modulation on human monocytes and neutrophils, with up- and down-regulation depending on the receptor. R- and S-form LPS activate human neutrophils similarly, despite the low CD14 expression, if the stimulation occurs in whole blood.

Do whole-body vibration exercise and resistance exercise modify concentrations of salivary cortisol and immunoglobulin A?

A single bout of resistance exercise (RE) induces hormonal and immune responses, playing an important role in a long-term adaptive process. Whole-body vibration (WBV) has also been shown to affect hormonal responses. Evidence suggests that combining WBV with RE may amplify hormonal and immune responses due to the increased neuromuscular load. Therefore, the aim of this study was to evaluate salivary cortisol (Scortisol) and salivary IgA (SIgA) concentrations following a RE session combined or not with WBV. Nine university students (22.9 ± 5.1 years, 175.8 ± 5.2 cm, and 69.2 ± 7.3 kg) performed five sets of squat exercise (70% one-repetition-maximum) combined (R+V30) or not (R) with WBV at 30 Hz. Saliva samples were obtained before and after exercise. Subjects also rated their effort according to the Borg CR-10 scale (RPE). Data were analyzed by a mixed model. RPE was higher after R+V30 (8.3 ± 0.7) compared to R (6.2 ± 0.7). However, Scortisol (pre: 10.6 ± 7.6 and 11.7 ± 7.6, post: 8.3 ± 6.3 and 10.2 ± 7.2 ng/mL for R and R+V30, respectively) and SIgA concentrations (pre: 98.3 ± 22.6 and 116.1 ± 51.2, post: 116.6 ± 64.7 and 143.6 ± 80.5 µg/mL for R and R+V30, respectively) were unaffected. No significant correlations were observed between Scortisol and RPE (r = 0.45, P = 0.22; r = 0.30, P = 0.42, for R and R+V30, respectively). On the basis of these data, neither protocol modified salivary cortisol or IgA, although RPE was higher after R+V30 than R.

Whole-body vibration decreases the proliferativeb response of TCD4+ cells in elderly individuals with knee osteoarthritis

The aim of this study was to investigate the effect of adding whole-body vibration (WBV; frequency = 35 to 40 Hz; amplitude = 4 mm) to squat training on the T-cell proliferative response of elderly patients with osteoarthritis (OA) of the knee. This study was a randomized controlled trial in which the selected variables were assessed before and after 12 weeks of training. Twenty-six subjects (72 ± 5 years of age) were divided into three groups: 1) squat training with WBV (WBV, N = 8); 2) squat training without WBV (N = 10), and 3) a control group (N = 8). Women who were ≥60 years of age and had been diagnosed with OA in at least one knee were eligible. The intervention consisted of 12 uninterrupted weeks of squatting exercise training performed 3 times/week. Peripheral blood mononuclear cells were obtained from peripheral blood collected before and after training. The proliferation of TCD4+ and TCD8+ cells was evaluated by flow cytometry measuring the carboxyfluorescein succinimidyl ester fluorescence decay before and after the intervention (∆). The proliferative response of TCD4+ cells (P = 0.02, effect size = 1.0) showed a significant decrease (23%) in the WBV group compared to the control group, while there was no difference between groups regarding the proliferative response of TCD8+ cells (P = 0.12, effect size = 2.23). The data suggest that the addition of WBV to squat exercise training might modulate T-cell-mediated immunity, minimizing or slowing disease progression in elderly patients with OA of the knee.

Whole transcriptome organisation in the dehydrated supraoptic nucleus

The supraoptic nucleus (SON) is part of the central osmotic circuitry that synthesises the hormone vasopressin (Avp) and transports it to terminals in the posterior lobe of the pituitary. Following osmotic stress such as dehydration, this tissue undergoes morphological, electrical and transcriptional changes to facilitate the appropriate regulation and release of Avp into the circulation where it conserves water at the level of the kidney. Here, the organisation of the whole transcriptome following dehydration is modelled to fit Zipf's law, a natural power law that holds true for all natural languages, that states if the frequency of word usage is plotted against its rank, then the log linear regression of this is -1. We have applied this model to our previously published euhydrated and dehydrated SON data to observe this trend and how it changes following dehydration. In accordance with other studies, our whole transcriptome data fit well with this model in the euhydrated SON microarrays, but interestingly, fit better in the dehydrated arrays. This trend was observed in a subset of differentially regulated genes and also following network reconstruction using a third-party database that mines public data. We make use of language as a metaphor that helps us philosophise about the role of the whole transcriptome in providing a suitable environment for the delivery of Avp following a survival threat like dehydration.

Ropinirole alters gene expression profiles in SH-SY5Y cells: a whole genome microarray study

Ropinirole (ROP) is a dopamine agonist that has been used as therapy for Parkinson's disease. In the present study, we aimed to detect whether gene expression was modulated by ROP in SH-SY5Y cells. SH-SY5Y cell lines were treated with 10 µM ROP for 2 h, after which total RNA was extracted for whole genome analysis. Gene expression profiling revealed that 113 genes were differentially expressed after ROP treatment compared with control cells. Further pathway analysis revealed modulation of the phosphatidylinositol 3-kinase (PI3K) signaling pathway, with prominent upregulation of PIK3C2B. Moreover, batches of regulated genes, including PIK3C2B, were found to be located on chromosome 1. These findings were validated by quantitative RT-PCR and Western blot analysis. Our study, therefore, revealed that ROP altered gene expression in SH-SY5Y cells, and future investigation of PIK3C2B and other loci on chromosome 1 may provide long-term implications for identifying novel target genes of Parkinson's disease.

Determining economical TBHQ doses for corn oil stability

Corn oil obtained from a Brazilian industry, free from antioxidants and citric acid, added of different tertiary butylhydroquinone (TBHQ) concentrations, was submitted to accelerated oxidation in the Schaal oven test at 63°C for 120 hours and for 168 hours in a photooxidation chamber. Peroxide and absorptivity values at 232nm and 270nm were determined for this oil. From the Schaal oven test results, the best and the economical TBHQ doses were determined to this oil. Afterwards, a shelf life experiment was conducted and confirmed 115mg.kg-1 TBHQ as the best and economical dose for that oil.

Evaluation of the in vitro glycemic index of a fiber-rich extruded breakfast cereal produced with organic passion fruit fiber and corn flour

The aim of this study was to determine the influence of process parameters and Passion Fruit Fiber (PFF) addition on the Glycemic Index (GI) of an extruded breakfast cereal. A 2³ Central Composite Rotational Design (CCRD) was used, with the following independent variables: raw material moisture content (18-28%), 2nd and 3rd barrel zone temperatures (120-160 ºC), and PFF (0-30%). Raw materials (organic corn flour and organic PFF) were characterized as to their proximate composition, particle size, and in vitro GI. The extrudates were characterized as to their in vitro GI. The Response Surface Methodology (RSM) and Principal Component Analysis (PCA) were used to analyze the results. Corn flour and PFF presented 8.55 and 7.63% protein, 2.61 and 0.60% fat, 0.52 and 6.17% ash, 78.77 and 78.86% carbohydrates (3 and 64% total dietary fiber), respectively. The corn flour particle size distribution was homogeneous, while PFF presented a heterogeneous particle size distribution. Corn flour and PFF presented values of GI of 48 and 45, respectively. When using RSM, no effect of the variables was observed in the GI of the extrudates (average value of 48.41), but PCA showed that the GI tended to be lower when processing at lower temperatures (<128 ºC) and at higher temperatures (>158 ºC). When compared to white bread, the extrudates showed a reduction of the GI of up to 50%, and could be considered an interesting alternative in weight and glycemia control diets.

Sensory evaluation and nutritional value of cakes prepared with whole flaxseed flour

The objective of this study was to evaluate the nutritional value, the oxidative stabilitiy, and consumer acceptance of cakes containing four different concentrations of flaxseed flour (5, 15, 30 and 45%) as partial replacement for wheat flour. The oxidative stability of polyunsaturated fatty acids was evaluated through the lipid peroxidation test (TBARS) in the flour and cakes. Linolenic acid was determined by gas chromatography as well as contents of protein, lipid, ash, and dietary fiber. Consumer acceptance was assessed using a structured hedonic scale of nine points. The oxidative stability of lipid flaxseeds was not affected by the heat treatment during flour processing and cake baking. Cakes made with 5, 15, and 30% of flaxseed flour, the most accepted by consumers, had dietary fiber levels ranging from 3.5 to 6.2 g and linolenic acid ranging from 445 to 2,500 mg.100 g-1 of the product. The cakes received claims of good and excellent source of dietary fiber and linolenic acid, respectively, both are bioactive compounds. The use of up to 30% of flaxseed flour in the preparation of cakes is a useful strategy to optimize the consumption of food rich in functional ingredients.

Rheological behavior of corn and soy mix as feed ingredients

Foods behave as non-Newtonian fluids, but little is known about how corn and soybean mix behave under viscometric flow. In order to characterize the rheological behavior of animal feed under viscometric flow, a 70:30 (mass:mass) mixture of ground corn and soybean grains was submitted to a capillary rheometer at 3 different temperatures (80, 120, and 160 °C), different moisture levels (26.5 ± 0.08; 30.4 ± 0.31, and 33.4 ± 0.05%), and 4 shear rates (30.4; 72.9; 304.3, and 728.6/second). Different strain rates and die dimensions were used to obtain the target shear rates. The resulting data were fitted to Power Law, Casson, and Bingham models. Based on experimental data, water content, mass temperature, and the effects of shear rate on the apparent shear viscosity of corn-soy mix were fitted to a single expression (p < 0.001, R² = 0.93): η = 18,769.7 (y)-0.86 e (-9.34 U + 935 T), where y is shear rate, U is sample moisture, and T is sample temperature in Kelvin scale. As expected, such mixture presented a pseudoplastic (shear-thinning) behavior.

Production and partial characterization of alkaline polygalacturonase secreted by thermophilic Bacillus sp. SMIA-2 under submerged culture using pectin and corn steep liquor

Polygalacturonase production by the thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.5% (w/v) apple pectin and supplemented with 0.3% (w/v) corn steep liquor, reached its maximum after 36 hours with levels of 39 U.mL-1. The increase in apple pectin and corn steep liquor concentrations in the medium from 0.5 and 0.3%, respectively, to 0.65%, markedly affected the production of polygalacturonase, whose activity increased four times, reaching a maximum of 150.3 U.mL-1. Studies on polygalacturonase characterization revealed that the optimum temperature of this enzyme was between 60-70 °C. Thermostability profile indicated that the enzyme retained about 82 and 63% of its activity at 60 and 70 °C, respectively, after 2 hours of incubation. The optimum pH of the enzyme was found to be 10.0. After incubation of crude enzyme solution at room temperature for 2 hours at pH 8.0, a decrease of about 29% on its original activity was observed. At pH 10.0, the decrease was 25%.

Color and chemical composition and of green corn produced under organic and conventional conditions

The present study aimed to evaluate the chemical properties of green corn, grown in both organic and conventional farming systems, using a completely randomized factorial design. Four corn varieties (AG 1051, BR 106, SWB 551 and VIVI) of green corn kernels were evaluated for color, proximate composition, total calories, carotenoids and bioactive amines. The farming system affected some chemical and physical characteristics of green corn, but this effect was dependent upon variety. In general, organic green corn kernels were reddish (a* color component) and had higher levels of β-carotene compared to the conventional ones, suggesting that these characteristics are related. Moreover, organic green corn had higher levels of total carbohydrates and total energy compared to conventional varieties. On the other hand, crude fiber levels were higher in conventional grains - an unexpected result that deserves further investigation. Finally, the levels of cadaverine and spermine bioactive amines were not affected either by the corn variety or by the farming system.

Isotope analysis (δ13C) of pulpy whole apple juice

The objectives of this study were to develop the method of isotope analysis to quantify the carbon of C3 photosynthetic cycle in pulpy whole apple juice and to measure the legal limits based on Brazilian legislation in order to identify the beverages that do not conform to the Ministry of Agriculture, Livestock and Food Supply (MAPA). This beverage was produced in a laboratory according to the Brazilian law. Pulpy juices adulterated by the addition of sugarcane were also produced. The isotope analyses measured the relative isotope enrichment of the juices, their pulpy fractions (internal standard) and purified sugar. From those results, the quantity of C3 source was estimated by means of the isotope dilution equation. To determine the existence of adulteration in commercial juices, it was necessary to create a legal limit according to the Brazilian law. Three brands of commercial juices were analyzed. One was classified as adulterated. The legal limit enabled to clearly identify the juice that was not in conformity with the Brazilian law. The methodology developed proved efficient for quantifying the carbon of C3 origin in commercial pulpy apple juices.

Simultaneous α-amylase and protease production by the soil bacterium Bacillus sp. SMIA-2 under submerged culture using whey protein concentrate and corn steep liquor: compatibility of enzymes with commercial detergents

Protease and α-amylase production by a thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.25% (w/v) starch as a carbon source reached a maximum at 18 hours (47 U.mg-1 Protein) and 36 hours (325 U.mg-1 Protein), respectively. Culture medium supplementation with whey protein concentrate (0.1%, w/v) and corn steep liquor (0.3%, w/v) not only improved the production of both enzymes but also enabled them to be produced simultaneously. Under these conditions, α-amylase and protease production reached a maximum in 18 hours with levels of 401 U.mg-1 protein and 78 U.mg-1 protein, respectively. The compatibility of the enzymes produced with commercial laundry detergent was investigated. In the presence of Campeiro® detergent, α-amylase activity increased while protease activity decreased by about 27%. These enzymes improved the cleaning power of Campeiro® detergent since they were able to remove egg yolk and tomato sauce stains when used in this detergent.