Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

Exercise training in the aerobic/anaerobic metabolic transition prevents glucose intolerance in alloxan-treated rats

Background: Ninety percent of cases of diabetes are of the slowly evolving non-insulin-dependent type, or Type 2 diabetes. Lack of exercise is regarded as one of the main causes of this disorder. In this study we analyzed the effects of physical exercise on glucose homeostasis in adult rats with type 2 diabetes induced by a neonatal injection of alloxan. Methods: Female Wistar rats aged 6 days were injected with either 250 mg/ kg of body weight of alloxan or citrate buffer 0.01 M (controls). After weaning, half of the animals in each group were subjected to physical training adjusted to meet the aerobic-anaerobic metabolic transition by swimming 1 h/day for 5 days a week with weight overloads. The necessary overload used was set and periodically readjusted for each rat through effort tests based on the maximal lactate steady state procedure. When aged 28, 60, 90, and 120 days, the rats underwent glucose tolerance tests (GTT) and their peripheral insulin sensitivity was evaluated using the HOMA index. Results: The area under the serum glucose curve obtained through GTT was always higher in alloxan-treated animals than in controls. A decrease in this area was observed in trained alloxan-treated rats at 90 and 120 days old compared with non-trained animals. At 90 days old the trained controls showed lower HOMA indices than the non-trained controls. Conclusion: Neonatal administration of alloxan induced a persistent glucose intolerance in all injected rats, which was successfully counteracted by physical training in the aerobic/anaerobic metabolic transition. © 2008 Mota et al; licensee BioMed Central Ltd.

Exercise at anaerobic threshold intensity and insulin secretion by isolated pancreatic islets of rats

To evaluate the effect of acute exercise and exercise training at the anaerobic threshold (AT) intensity on aerobic conditioning and insulin secretion by pancreatic islets, adult male Wistar rats were submitted to the lactate minimum test (LMT) for AT determination. Half of the animals were submitted to swimming exercise training (trained), 1 h/day, 5 days/week during 8 weeks, with an overload equivalent to the AT. The other half was kept sedentary. At the end of the experimental period, the rats were submitted to an oral glucose tolerance test and to another LMT. Then, the animals were sacrificed at rest or immediately after 20 minutes of swimming exercise at the AT intensity for pancreatic islets isolation. At the end of the experiment mean workload (% bw) at AT was higher and blood lactate concentration (mmol/L) was lower in the trained than in the control group. Rats trained at the AT intensity showed no alteration in the areas under blood glucose and insulin during OGTT test. Islet insulin content of trained rats was higher than in the sedentary rats while islet glucose uptake did not differ among the groups. The static insulin secretion in response to the high glucose concentration (16.7 mM) of the sedentary group at rest was lower than the sedentary group submitted to the acute exercise and the inverse was observed in relation to the trained groups. physical training at the AT intensity improved the aerobic condition and altered insulin secretory pattern by pancreatic islets. © 2010 Landes Bioscience.

Comparison between protocols for the determination of anaerobic threshold in adolescent swimmers

Aim. The purpose of this study was to compare the anaerobic threshold speed (AT) obtained from fixed lactate blood concentrations (AT 4 mM and AT 3.5 mM), lactate minimum speed (LM) and critical speed (CS), determined from different distances in fifteen Brazilian national level swimmers (10 boys = 14.8 ± 0.6 years old and 5 girls = 14.6 ±0.8 year-old). Methods. The tests to determine the AT 4 mM, AT 3.5 mM, LM and CS were performed in a 25 m swimming pool and consisted of 7 or 8 evaluations separated by 24-48 h intervals. Data were submitted to analysis of variance (ANOVA) for repeated measures, followed by the post hoc Scheffé test and Pearson correlation coefficients. Significance was set at P<0.01. Results. There were no significant differences among the values for AT 4 mM and CS1 (1.34 ± 0.05 vs. 1.33 ± 0.05 m.s -1, respectively). However, AT 4 mM and CS1 were significantly higher than AT 3.5 mM (1.28 ± 0.04 m.s -1), LM (1.27 ± 0.05 m.s -1), CS2 (1.26 ± 0.06 m.s -1), CS3 (1.27 ± 0.06 m.s -1) and CS4 (1.25 ± 0.07 m.s -1). There were no significant differences among the values for AT 3.5 mM, LM, CS2, CS3 and CS4. Conclusion. The results obtained in this study suggest that the anaerobic threshold determined by a fixed lactate concentration of 3.5 mM, as well as the LM and the CS methods determined by different distances, seem to be the most appropriate indexes for the evaluation of aerobic capacity in adolescent swimmers.

Simple electroanalytical method for determination of guanosine in biological sample

A poly glutamic acid film modified electrode exhibited a catalytic response toguanosine oxidation potential and higher peak current value. Linear concentration curve was obtained in the concentration interval of 1.0 a 10.0 μmol L-1 in 0.04 mol L-1 B-R buffer pH 2.0 with a detection limit of 0.198 μmol L-1. The electrode was used for the determination of guanosine in the potential of +1.1 V (vs. Ag/AgCl) using differential pulse voltammetry (DPV) at urine sample with good recovery. © 2010 by CEE.

Stroke phases responses around maximal lactate steady state in front crawl

The objective of this study was to analyze changes in stroke rate (SR), stroke length (SL) and stroke phases (entry and catch, pull, push and recovery) when swimming at (MLSS) and above (102.5% MLSS) the maximal lactate steady state. Twelve endurance swimmers (21±8 year, 1.77±0.10m and 71.6±7.7kg) performed in different days the following tests: (1) 200- and 400-m all-out tests, to determine critical speed (CS), and; (2) 2-4 30-min sub-maximal constant-speed tests, to determine the MLSS and 102.5% MLSS. There was significant difference among MLSS (1.22±0.05ms-1), 102.5% MLSS (1.25±0.04ms-1) and CS (1.30±0.08ms-1). SR and SL were maintained between the 10th and 30th minute of the test swum at MLSS and have modified significantly at 102.5% MLSS (SR - 30.9±3.4 and 32.2±3.5cyclesmin-1 and SL - 2.47±0.2 and 2.38±0.2mcycle-1, respectively). All stroke phases were maintained at 10th and 30th minute at MLSS. However, the relative duration of propulsive phase B (pull) increased significantly at 102.5% MLSS (21.7±3.4% and 22.9±3.9%, respectively). Therefore, the metabolic condition may influence the stroke parameters (SR and SL) and stroke strategy to maintain the speed during swim tests lasting 30min. © 2010 Sports Medicine Australia.

Non-exhaustive test for aerobic capacity determination in running rats

A simple and applicable method for non-exhaustive aerobic evaluation in running rats is described. Wistar rats were submitted to running test at different velocities (10, 15, 20, 25 m/min) with 48 h recovery among them. At each velocity, the rats ran two bouts of 5 min with 2 min of rest between bouts. Blood samples were collected at the end of each bout for lactate determination. For each intensity, delta lactate was calculated and using deltas obtained by four tests, an individual linear interpolation was plotted. The y-intercept of linear interpolation was the null delta lactate equivalent to the critical velocity (CV). To verify the lactate stabilization at CV, the animals were submitted to 25 min of continuous exercise (15, 20, 25 m/min), with blood collection every 5 min. The estimated CV was 16.6±0.7 m/min, with significant linear regressions (R=0.90±0.03). The rats presented maximal lactate steady state (MLSS) at 3.9±0.4 mmol/L, at 20 m/min. The CV was less than MLSS but significantly correlated with this parameter (r=0.78). This non-exhaustive test seems to be valid for the aerobic evaluation of sedentary rats and this protocol underestimates the MLSS in 20%. This test seems to be the interesting method for the evaluation of rats submitted to acute exercise or physical training.

Perfil Fermentativo e Valor Nutritivo de Silagem de Capim-Elefante Inoculada com Streptococcus Bovis

The experiment was developed to evaluate the effect of strains of Streptococcus bovis (HC5 and JB1) on pH, ammonia production and nutritional value in elephant-grass silages (Pennisetum purpureum Schum). The experimental design was entirely randomized, with three treatments: T1 - elephant- grass, T2 - grass-elephant inoculated with Streptococcus bovis HC5, T3 - elephant grass inoculated with Streptococcus bovis JB1, totaling five replicates per treatment. The smallest value of ammonia was observed in the silage treated with inoculante the base Streptococcus bovis JB1 (5.90% N-total). Inoculation increased the levels of lactic acid and decreased levels of acetic, propionic and butyric acids, and the silages inoculated with Streptococcus bovis JB1 and HC5 were those with the highest values of lactic acid. There were not statistical differences for the tenors of dry matter (MS) and crude protein (PB). For the values of the fibers, the grass-elephant silage inoculated with Streptococcus bovis JB1, it presented the smallest value of neutral detergent fiber (59.77 %) and Hemicellulose (26.71 %). The largest value of in vitro dry matter digestibility (DIVMS) was observed in the silage with Streptococcus bovis JB1, showing superiority of four percentile points (64.23 %) in relation to the treatment without bacterial inoculante (60.35 %). The use Streptococcus bovis JB1 was efficient in the process fermentativo besides promoting would get better in the quality of the fiber and DIVMS in the studied silage.

Aerobic fitness evaluation during walking tests identifies the maximal lactate steady state

Objective. The aim of this study was to verify the possibility of lactate minimum (LM) determination during a walking test and the validity of such LM protocol on predicting the maximal lactate steady-state (MLSS) intensity. Design. Eleven healthy subjects (24.2 ± 4.5 yr; 74.3 ± 7.7 kg; 176.9 ± 4.1 cm) performed LM tests on a treadmill, consisting of walking at 5.5 km h -1 and with 20-22% of inclination until voluntary exhaustion to induce metabolic acidosis. After 7 minutes of recovery the participants performed an incremental test starting at 7% incline with increments of 2% at each 3 minutes until exhaustion. A polynomial modeling approach (LMp) and a visual inspection (LMv) were used to identify the LM as the exercise intensity associated to the lowest [bLac] during the test. Participants also underwent to 24 constant intensity tests of 30 minutes to determine the MLSS intensity. Results. There were no differences among LMv (12.6 ± 1.7 %), LMp (13.1 ± 1.5 %), and MLSS (13.6 ± 2.1 %) and the Bland and Altman plots evidenced acceptable agreement between them. Conclusion. It was possible to identify the LM during walking tests with intensity imposed by treadmill inclination, and it seemed to be valid on identifying the exercise intensity associated to the MLSS. Copyright © 2012 Guilherme Morais Puga et al.

Endurance training does not change the relative intensity at lactate threshold in elderly

Aim. The aim of the study was to verify whether endurance training may induce changes on the percentage of peak heart rate (% peak HR) at the lactate threshold (LT) intensity in untrained elderly. Methods. Sixteen healthy men (64.3 ± 4.1 yrs) underwent an incremental test on cycloergometer to determine the LT and the corresponding % peak HR at LT intensity. Afterwards, they were randomly distributed into two groups (n = 8 each): endurance training (ET) and control (C). The ET exercised 3 days a week for 12 weeks. The training session was divided into warm-up (5 min at 50% of LT;), a main part, and a cool-down (5 min 50% below of LT). The main part had a gradual increased volume through the weeks of 2 min. The initial volume on the 1st week was 25 min reaching 47 min at the 12th week. The relative intensity was kept constant (90 to 100% of LT). Results. After 12 weeks, the % peak HR at LT did not change significantly for both groups P > 0.05 (ET 82.9 ± 4.1 vs. 82.5 ± 3.4 and Ç 80.2 ± 7.1 vs. 81.8 ± 7.1). Conclusion. We conclude that endurance training proposed does not change the relative intensity at LT in elderly.

Efeitos do extrato de parede de levedura em dieta seca sobre a Microbiologia, ácidos graxos de cadeia curta e redução do odor das fezes de gatos adultos

The aim of this study was to evaluate the effects of yeast cell wall extract (YCW) in dry diet on the fecal microbiota, concentration of short-chain fatty acids (SCFA) and on the odor reduction of cats feces. We used 20 animals of both sexes, randomly assigned to four treatments and five repetitions totaling 20 experimental units: 1) dry commercial diet (control); 2) control + 0.2%, 3) control + 0.4%, and 4) control + 0.6% of YCW in dry matter. Enterobacteriaceae and lactic acid bacteria, fecal concentration of acetic, propionic and butyric acids, ammonia nitrogen and sensory panel were performed. There were no significant differences (p> 0.05) for bacterial counts and the concentration of SCFA and ammonia, but in sensory panel a reduction in the odor of feces could be noted with the use of 0.2% of YCW. We concluded that the addition of up to 0.6% YCW had no effect on the microbiology and the concentration of fatty acids, but there is potential for its use as an additive because of the improvement in the odor of feces. However, further studies are needed to understand the mechanisms of action and the effects of prebiotics for domestic cats.

Monitoring chronic physical stress using biomarkers, performance protocols and mathematical functions to identify physiological adaptations in rats

This study was undertaken to characterize the effects of monotonous training at lactate minimum (LM) intensity on aerobic and anaerobic performances; glycogen concentrationsin the soleus muscle, the gastrocnemius muscle and the liver; and creatine kinase (CK), free fatty acids and glucose concentrations in rats. The rats were separated into trained (n =10), baseline (n = 10) and sedentary (n=10) groups. The trained group was submitted to the following: 60 min/day, 6 day/week and intensity equivalent to LM during the 12-week training period. The training volume was reduced after four weeks according to a sigmoid function. The total CK (U/L) increased in the trained group after 12 weeks (742.0±158.5) in comparison with the baseline (319.6±40.2) and the sedentary (261.6+42.2) groups. Free fatty acids and glycogen stores (liver, soleus muscle and gastrocnemius muscle) increased after 12 weeks of monotonous training but aerobic and anaerobic performances were unchanged in relation to the sedentary group. The monotonous training at LM increased the level of energy substrates, unchanged aerobic performance, reduced anaerobic capacity and increased the serum CK concentration; however, the rats did not achieve the predicted training volume.

Moderate physical activity from childhood contributes to metabolic health and reduces hepatic fat accumulation in adult rats

Background: Obesity, oxidative stress and inflammation, by triggering insulin resistance, may contribute to the accumulation of hepatic fat, and this accumulation by lipotoxicity can lead the organ to fail. Because obesity is growing at an alarming rate and, worryingly, in a precocious way, the present study aimed to investigate the effects of moderate physical training performed from childhood to adulthood on liver fat metabolism in rats. Methods. Twenty rats that were 28days old were divided into two groups: control (C) and trained (T). The C Group was kept in cages without exercise, and the T group was submitted to swimming exercise for 1hour/day, 5days/week from 28 to 90days of age (8weeks) at 80% of the anaerobic threshold determined by the lactate minimum test. At the end of the experiment, the body weight gain, insulin sensitivity (glucose disappearance rate during the insulin tolerance test), concentrations of free fatty acids (FFA) and triglycerides (TG) and hepatic lipogenic rate were analyzed. For the statistical analysis, the Student t-test was used with the level of significance preset at 5%. Results: The T group showed lower body weight gain, FFA concentrations, fat accumulation, hepatic lipogenic rate and insulin resistance. Conclusion: The regular practice of moderate physical exercise from childhood can contribute to the reduction of obesity and insulin resistance and help prevent the development of accumulation of hepatic fat in adulthood. © 2013de Moura et al; licensee BioMed Central Ltd.

Sugarcane bagasse ozonolysis pretreatment: Effect on enzymatic digestibility and inhibitory compound formation

Sugarcane bagasse was pretreated with ozone to increase lignocellulosic material digestibility. Bagasse was ozonated in a fixed bed reactor at room temperature, and the effect of the two major parameters, ozone concentration and sample moisture, was studied. Acid insoluble and total lignin decreased whereas acid soluble lignin increased in all experiments. Pretreatment barely attacked carbohydrates, with cellulose and xylan recovery rates being >92%. Ozonolysis increased fermentable carbohydrate release considerably during enzymatic hydrolysis. Glucose and xylose yields increased from 6.64% and 2.05%, for raw bagasse, to 41.79% and 52.44% under the best experimental conditions. Only xylitol, lactic, formic and acetic acid degradation compounds were found, with neither furfural nor HMF (5-hydroxymethylfurfural) being detected. Washing detoxification provided inhibitor removal percentages above 85%, increasing glucose hydrolysis, but decreasing xylose yield by xylan solubilization. SEM analysis showed structural changes after ozonization and washing. © 2013 Elsevier Ltd.

Lactobacillus acidophilus CRL 1014 improved gut health in the SHIME® reactor

Background: How to maintain gut health is a goal for scientists throughout the world. Therefore, microbiota management models for testing probiotics, prebiotics, and synbiotics have been developed.Methods: The SHIME® model was used to study the effect of Lactobacillus acidophilus 1014 on the fermentation pattern of the colon microbiota. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 2-wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota was monitored. The microbiota was then subjected to a 4-wk treatment period by adding 5 mL of sterile peptone water with L. acidophilus CRL1014 at the concentration of 108 CFU/mL to vessel one (the stomach compartment). Plate counts, Denaturing Gradient Gel Electrophoresis (DGGE), short-chain fatty acid (SCFA) and ammonium analyses were carried out for monitoring of the microbial community from the colon compartments.Results: A significant increase (p < 0.01) in the Lactobacillus spp. and Bifidobacterium spp. populations was observed during the treatment period. The DGGE obtained showed changes in the lactobacilli community from the colon compartments of the SHIME® reactor. The (SCFA) concentration increased (p < 0.01) during the treatment period, due mainly to significant increased levels of acetic, butyric, and propionic acids. However, ammonium concentrations decreased during the same period (p < 0.01).Conclusions: This study showed the beneficial influence of L. acidophilus CRL 1014 on microbial metabolism and lactobacilli community composition for improving human health. © 2013 Sivieri et al.; licensee BioMed Central Ltd.