Hypochlorous acid inhibition by acetoacetate: Implications on neutrophil functions

Type-1 diabetic patients experience hyperketonemia caused by an increase in fatty acid metabolism. Thus, the aim of this study was to measure the effect of ketone bodies as suppressors of oxidizing species produced by stimulated neutrophils. Both acetoacetate and 3-hydroxybutyrate have suppressive effect on the respiratory burst measured by luminol-enhanced chemiluminescence. Through measurements of hypochlorous acid production, using neutrophils or the myeloperoxidase/H2O2/Cl- system, it was found that acetoacetate but not 3-hydroxybutyrate is able to inhibit the generation of this antimicrobial oxidant. The superoxide anion scavenging properties were confirmed by ferricytochrome C reduction and lucigenin-enhanced chemiluminescence assays. However, ketone bodies did not alter the rate of oxygen uptake by stimulated neutrophils, measured with an oxygen electrode. A strong inhibition of the expression of the cytokine IL-8 by cultured neutrophils was also observed; this is discussed with reference to the antioxidant-like property of acetoacetate. © 2004 Pharmaceutical Society of Japan.

Fast preparative separation of flavones from capitula of Eriocaulon ligulatum (Vell.) L.B.Smith (Eriocaulaceae) by High-speed countercurrent chromatography (HSCCC)

High-speed countercurrent chromatography (HSCCC) is a leading method for the fast separation of natural products from plants. It was used for the preparative isolation of two flavone monoglucosides present in the capitula of Eriocaulon ligulatum (Veil.) L.B.Smith (Eriocaulaceae). This species, known locally as botão-dourado, is exported to Europe, Japan and North America as an ornamental species, constituting an important source of income for the local population of Minas Gerais State, Brazil. The solvent system, optimized in tests prior to the HSCCC run, consisted of the two phases of the mixture ethyl acetate: n-propanol: water (140:8:80, v/v/v), which led to the successful separation of 6-methoxyluteolin-7-O-β-D-allopyranoside and 6-methoxyapigenin-7-O-β-D-allopyranoside in only 3 hours. The two flavonoids were identified by NMR (1-D and 2-D) and ESI-MS, comparing their spectra with published data.

Microhardness and fluoride release of restorative materials in different storage media

This study evaluated the surface microhardness and fluoride release of 5 restorative materials - Ketac-Fil Plus, Vitremer, Fuji II LC, Freedom and Fluorofil - in two storage media: distilled/deionized water and a pH-cycling (pH 4.6). Twelve specimens of each material, were fabricated and the initial surface microhardness (ISM) was determined in a Shimadzu HMV-2000 microhardness tester (static load Knoop). The specimens were submitted to 6- or 18-h cycles in the tested media. The solutions were refreshed at the end of each cycle. All solutions were stored for further analysis. After 15-day storage, the final surface microhardness (FSM) and fluoride release were measured. Fluoride dose was measured with a fluoride-specific electrode (Orion 9609-BN) and digital ion analyzer (Orion 720 A). The variables ISM, FSM and fluoride release were analyzed statistically by analysis of variance and Tukey's test (p<0.05). There was significant difference in FSM between the storage media for Vitremer (pH 4.6 = 40.2 ± 1.5; water = 42.6 ± 1.4), Ketac-Fil Plus (pH 4.6 = 73.4 ± 2.7; water = 58.2 ± 1.3) and Fluorofil (pH 4.6 = 44.3 ± 1.8; water = 38.4 ± 1.0). Ketac-Fil Plus (9.9 ± 18.0) and Fluorofil (4.4 ± 1.3) presented higher fluoride release in water, whereas Vitremer (7.4 ± 7.1), Fuji II LC (5.7 ± 4.7) and Freedom (2.1 ± 1.7) had higher fluoride release at pH 4.6. Microhardness and fluoride release of the tested restorative materials varied according to the storage medium.

Effectiveness of six different disinfectants on removing five microbial species and effects on the topographic characteristics of acrylic resin

Purpose: The aim of this study was to evaluate the effectiveness of disinfectant solutions (1% sodium hypochlorite, 2% chlorhexidine digluconate, 2% glutaraldehyde, 100% vinegar, tabs of sodium perborate-based denture cleanser, and 3.8% sodium perborate) in the disinfection of acrylic resin specimens (n = 10/group) contaminated in vitro by Candida albicans, Streptococcus mutans, S. aureus, Escherichia coli, or Bacillus subtilis as measured by residual colony-forming unit (CFU). In a separate experiment, acrylic resin was treated with disinfectants to monitor potential effects on surface roughness, Ra (μm), which might facilitate microbial adherence. Materials and Methods: Three hundred fifty acrylic resin specimens contaminated in vitro with 1×10 6 cells/ml suspensions of standard strains of the cited microorganisms were immersed in the disinfectants for 10 minutes; the control group was not submitted to any disinfection process. Final counts of microorganisms per ml were performed by plating method for the evaluation of microbial level reduction. Results were compared statistically by ANOVA and Tukey's test (p ≤ 0.05). In a parallel study aiming to evaluate the effect of the tested disinfectant on resin surface, 60 specimens were analyzed in a digital rugosimeter before and after ten cycles of 10-minute immersion in the disinfectants. Measurements of superficial roughness, Ra (μm), were compared statistically by paired t-test (p ≤ 0.05). Results: The results showed that 1% sodium hypochlorite, 2% glutaraldehyde, and 2% chlorhexidine digluconate were most effective against the analyzed microorganisms, followed by 100% vinegar, 3.8% sodium perborate, and tabs of sodium perborate-based denture cleanser. Superficial roughness of the specimens was higher after disinfection cycles with 3.8% sodium perborate (p = 0.03) and lower after the cycles with 2% chlorhexidine digluconate (p = 0.04). Conclusion: Within the limits of this experiment, it could be concluded that 1% sodium hypochlorite, 2% glutaraldehyde, 2% chlorexidine, 100% vinegar, and 3.8% sodium perborate are valid alternatives for the disinfection of acrylic resin. © 2008 by The American College of Prosthodontists.

Carotenóides totais em Pothomorphe umbellata (L.) Miq. cultivadas in vitro

The aim of the present work was to compare the content of carotenoids between callus and regenerated plants of Pothomorphe umbellate. Germinated seeds (40 days old) were inoculated in different concentrations and combinations of BAP (benzylaminopurine) and NAA (naphthalene acetic acid) in order to stimulate the callus' production. After 60 days of culture, the callus containing some shoots were transferred to organogenesis medium (GA 3 0.1 mg L -1, BAP 0.5 mg L -1) for 40 days. Next, they were subcultivated in a medium for seedling growth (without regulators) for 40 days. Callus (collected after 60 days) and seedlings (collected after 140 days) were frozen in liquid nitrogen and kept under -80°C for future carotenoids' analysis. The highest concentration of carotenoids was found in plants cultivated in medium without regulators. The callus did not showed difference concerning the culture medium; however, they presented lower content of carotenoids in relation to plants.

Indoor air quality assessment of elementary schools in Curitiba, Brazil

The promotion of good indoor air quality in schools is of particular public concern for two main reasons: (1) school-age children spend at least 30% of their time inside classrooms and (2) indoor air quality in urban areas is substantially influenced by the outdoor pollutants, exposing tenants to potentially toxic substances. Two schools in Curitiba, Brazil, were selected to characterize the gaseous compounds indoor and outdoor of the classrooms. The concentrations of benzene, toluene, ethylbenzene, and the isomers xylenes (BTEX); NO2; SO2; O3; acetic acid (HAc); and formic acid (HFor) were assessed using passive diffusion tubes. BTEX were analyzed by gas chromatography-ion trap mass spectrometry and other collected gasses by ion chromatography. The concentration of NO2 varied between 9.5 and 23 μg m-3, whereas SO2 showed an interval from 0.1 to 4.8 μg m-3. Within the schools, BTEX concentrations were predominant. Formic and acetic acids inside the classrooms revealed intermediate concentrations of 1.5 μg m-3 and 1.2 μg m-3, respectively. © Springer Science + Business Media B.V. 2009.

La suplementación pre-ejercicio con carbohidrato de alta concentración perjudica el rendimiento en el ejercicio de alta intensidad en bicicleta

Objectives. To evaluate the effects of pre-exercise high concentration carbohydrate supplementation on performance, cardiovascular, metabolic and hormonal responses during high intensity cycling exercise. Method. Seven male cyclists (28.7 ± 5.4 years; 65.2 ± 4.7 kg body weight), who performed two continuous exercise trials under placebo (PLA) or carbohydrate (CHO) ingestion at a work rate of 80% VO 2max until exhaustion, participated in the study. The cyclists received 5 ml.kg-1 of a maltodextrin solution diluted at a concentration of 10% (CHO) or placebo (PLA) at 60, 45 and 30 min pre-exercise. Results. A 5.4% reduction in the time to exhaustion was observed in the CHO trial compared to the PLA trial. In both trials, glucose and lactate levels were higher in the post-trial condition compared to pre-exercise values (p < 0.05). Free fatty acid levels were lower in the CHO group than in the PLA group both before and after the trial (p < 0.05). Insulinemia was higher during the pre-trial in the CHO group (42.7 ± 3.6 μU.ml-1) compared to the PLA condition (11.8 ± 3.3 μU.ml-1) (p < 0.05), and even decreased to 23.8 ± 5.1 μU.ml-1 during exercise after CHO intake (p < 0.05). No significant differences in plasma cortisol were observed between the two trials (p > 0.05). Conclusions. Pre-exercise high concentration CHO supplementation resulted in impaired performance in high intensity cycling exercise and decreased free fatty acid levels. © 2010 Revista Andaluza de Medicina del Deporte.

Effects of the ethyl acetate fraction of Alchornea triplinervia on healing gastric ulcer in rats

Alchornea triplinervia (Spreng.) Muell. Arg (Euphorbiaceae) is a medicinal plant commonly used by people living in the Cerrado region of Brazil to treat gastrointestinal ulcers. We previously described the gastroprotective action of methanolic extract (ME) of Alchornea triplinervia and the ethyl acetate fraction (EAF) in increasing of prostaglandin E 2 (PGE 2) gastric levels in the mucosa. In this work we evaluated the effect of EAF in promoting the healing process in rats with acetic acid-induced gastric ulcers. In addition, toxicity was investigated during treatment with EAF. After 14 days of treatment with EAF, the potent stimulator of gastric cell proliferation contributed to the acceleration of gastric ulcer healing. Upon immunohistochemical analysis, we observed a pronounced expression of COX-2, mainly in the submucosal layer. The 14-day EAF treatment also significantly increased the number of neutrophils in the gastric mucosa regeneration area. The EAF induced angiogenesis on gastric mucosa, observed as an increase of the number of blood vessels supplying the stomach in rats treated with EAF. Oral administration for 14 days of the ethyl acetate fraction from Alchornea triplinervia accelerated the healing of gastric ulcers in rats by promoting epithelial cell proliferation, increasing the number of neutrophils and stimulation of mucus production. This fraction, which contained mainly phenolic compounds, contributed to gastric mucosa healing. © 2011 by the authors; licensee MDPI, Basel, Switzerland.

Simple method for culture of peripheral blood lymphocytes of Testudinidae.

We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes.

Advances in drug design based on the amino acid approach: Taurine analogues for the treatment of CNS diseases

Amino acids are well known to be an important class of compounds for the maintenance of body homeostasis and their deficit, even for the polar neuroactive aminoacids, can be controlled by supplementation. However, for the amino acid taurine (2-aminoethanesulfonic acid) this is not true. Due its special physicochemical properties, taurine is unable to cross the blood-brain barrier. In addition of injured taurine transport systems under pathological conditions, CNS supplementation of taurine is almost null. Taurine is a potent antioxidant and anti-inflammatory semi-essential amino acid extensively involved in neurological activities, acting as neurotrophic factor, binding to GABA A/glycine receptors and blocking the excitotoxicity glutamate-induced pathway leading to be a neuroprotective effect and neuromodulation. Taurine deficits have been implicated in several CNS diseases, such as Alzheimer's, Parkinson's, epilepsy and in the damage of retinal neurons. This review describes the CNS physiological functions of taurine and the development of new derivatives based on its structure useful in CNS disease treatment.&; 2012 by the authors; licensee MDPI, Basel, Switzerland.

Phosphorus solubilizing and iaa production activities in plant growth promoting rhizobacteria from brazilian soils under sugarcane cultivation

Plant Growth Promoting Rhizobacteria (PGPR) has been used as a biofertilizer, bringing benefits to agriculture as Phosphorus Solubilizing Bacteria (PSB), indole-acetic acid (IAA) producers, and with other activites. The goal of this report was the identification of PGPR from soils under sugarcane crops by 16S rRNA sequencing, and the evaluation of the ability of phosphorus solubilizing and IAA production by biological assays. The isolates of this work were obtained from three areas of sugarcane crop from São Paulo State, Brazil. All isolates came from rhizosphere soil, and in a total of 60 isolates just 10 have showed high ability in phosphorus solubilizing. The selection of PSB may be done by phenotypic and/or genotypic characterization. Among ten isolates Enterobacter sp. (FJ890899), Entrobacter homaechei subsp. verschuerennii (FJ890998), Burkholderia sp. (FJ890895), and Labrys portucalensis (FJ890891) were able to IAA production. © 2006-2012 Asian Research Publishing Network (ARPN).

Blood parameters of chicks from eggs injected with ascorbic acid and subjected to thermal stress

This study was aimed to verify if chicks from eggs injected with ascorbic acid and subjected to thermal stress would have higher immunity than chicks from incubation at thermoneutrality without injection of ascorbic acid. The parameters evaluated were temperature on oxygen saturation in hemoglobin, glucose, number of erythrocytes, hematocrit rate and number of hemoglobins of newly hatched male chicks, hatched from eggs injected with ascorbic acid (AA) and subjected to thermal stress during incubation. The experimental design was completely randomized in factorial scheme 5 (application levels of ascorbic acid) x 2 (incubation temperatures). The data were subjected to analysis of variance using the General Linear Model procedure (GLM) of SAS ®. For the parameters (number of erythrocytes, rate of hematrocit and values of hemoglobin), there was significant interaction (p <0.05) between treatments in egg and incubation temperatures. Analyzing the interactions for these parameters, it was observed that the application of 0% ascorbic acid in egg minimized the effect of heat stress when compared with treatment without injection. The application of ascorbic acid levels in eggs incubated under heat stress failed to maximize the immunity of newly hatched chicks. It is assumed that the increased liquid in the amniotic fluid, in those embryos injected with water, favored the lower heat conductance for these embryos, thus helping in their development in relation to immunity. Considering that hemoglobin is related to the transport of gases, these data suggest that increasing the concentration of AA solution inoculated may influence the respiratory rates of eggs.

Antioxidant and cytotoxic effects of crude extract, fractions and 4-nerolidylcathecol from aerial parts of pothomorphe umbellata L. (Piperaceae)

The crude ethanolic extract from aerial parts of Pothomorphe umbellata L. (Piperaceae) and fractions obtained by partitions sequentially among water-methanol, methylene chloride, and ethyl acetate, as well as the major constituent, 4-nerolidylcatechol, were, respectively, evaluated and evidenced for antioxidant and cytotoxic effects through fluorometric microplate and microculture tetrazolium assays in HL-60 cells. The crude ethanolic extract demonstrated the preeminent antioxidant activity (IC50 = 1.2 μg/mL) against exogenous cytoplasmic reactive oxygen species, followed by the water-methanolic (IC50 = 4.5 μg/mL), methylene chloride (IC 50 = 5.9 g/mL), ethyl acetate (IC50 = 8.0 g/mL), 4-nerolidylcatechol (IC50 = 8.6 g/mL), and the sterol fractions (IC50 > 12.5 μg/mL). Vitamin C, the positive control used in this assay, presented IC50 value equivalent to 1.7 μg/mL. 4-Nerolidylcatechol (IC50 = 0.4 μg/mL) and methylene chloride fraction (IC50 = 2.3 μg/mL) presented considerable cytotoxicity probably because of the presence of an o-quinone, an auto-oxidation by product of the catechol. Polar compounds, present in the ethanol extract, appear to increase the solubility and stability of the major active constituent, acting synergistically with 4-nerolidylcatechol, improving its pharmacokinetic parameters and increasing significantly its antioxidant activity which, in turn, suggests that the aqueous-ethanolic extract, used in folklore medicine, is safe and effective. © 2013 Andrey P. Lopes et al.

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.

Exposição in utero ao desregulador endocrino bisfenol A e ao agente quimiopreventivo indol-3-carbinol: efeitos sobra a morfogênese e a suscetibilidade à carcinogênese prostática

Pós-graduação em Biologia Geral e Aplicada - IBB