Aderência e formação do biofilme por Staphylococcus aureus ao polietilenotereftalato (PET) usado como embalagem para acondionamento de medicamentos e cosméticos

The polyethylene terephthalate (PET) is used for package drugs and cosmetics. The aim of this research was examine by scanning electron microscopy (SEM) a Staphylococcus aureus attachment and biofilm formation on a polyethylene terephthalate (PET) surface and hydrophobicity of S. aureus by adherence to hydrocarbons. A suspension of S. aureus was prepared in Mueller-Hinton broth and, coupons of polyethylene terephthalate were incubated for 30 minutes, two, 24 and 48 hours, 15 and 30 days. Afteron the coupons were removed and prepared for scanning electron microscope analysis. The attachment and biofilm formation was observed on the surfaces of PET. The SEM revealed adhesion and biofilm formation on PET surfaces. The hydrophobicity test classified S. aureus as hydrophobic.

Effects of acetylsalicylic acid and acetic acid solutions in VX2 carcinoma cells. In vitro analysis

Purpose: To analyze, in vitro, the effects of acetylsalicylic acid (aspirin) and acetic acid solutions on VX2 carcinoma cells in suspension and to examine the correlation between these effects and neoplastic cell death. Methods: The VX2 tumor cells (107 cells/ml) were incubated in solutions containing differing concentrations (2.5% and 5%) of either acetylsalicylic acid or acetic acid, or in saline solution (controls). Every five minutes, cell viability was tested (using the trypan blue test) and analyzed under light microscopy. Results: Tumor cell viability (in %) decreased progressively and, by 30 minutes, neoplastic cell death had occurred in all solutions. Conclusion: Based on this experimental model and the methodology employed, we conclude that these solutions cause neoplastic cell death in vitro.

Estudo biológico e biométrico dos estádios ninfais de Rhodnius prolixus Stäl, 1859 (Hemiptera, Reduviidae) sob condições laboratoriais

Eggs and nymphs originated from couples of Rhodnius prolixus obtained from nymphs of the 5th instar were used for biological cycle and biometric studies. The following biological cycle parameters were determined under a temperature of 28°C and relative humidity, varying between 52 - 94% : medium period of incubation: 13.01 days; rate of eggs eclosion: 77.6%; medium period of development of the 1st, 2nd, 3rd, 4 th, 5th instar nymphs : 19.33, 19.09, 20.38, 24.37, 38.14 days, respectively; percentual of deaths in the nymph instar: 26.70, 14.00, 18.26, 17.02, 35.47% respectively; percentual of changes per instar nymphs; 73.30, 86.03, 81.73, 82.97, 64.52%, respectively. Biometric measurements performed, showed that in all the instars the abdomen is the largest segment. In the four first instars, the head is larger than the thorax. In the fifth instar, the head and thorax present are about the same size.

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Background: Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils. Methods: Eosinophils were purified using a percoll gradient followed byimmunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry. Results: At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils. Conclusion: Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion. © 2008 Lintomen et al; licensee BioMed Central Ltd.

Diferentes procedimentos na determinação de indicadores internos para estimativa de produção fecal e fluxo duodenal de matéria seca em bovinos

Times of in situ incubation (144 and 288h) for determination of internal markers IADF and INDF and the effects of differents procedures (wash or not the nylon bag every 72h incubation) was evaluated in samples of diet, duodenal digesta and cattle feces. The duodenal flow dry matter and fecal production utilizing the internal markers to compare with the external marker chromium oxide there was estimated. The animals were fed with sorgum silage, concentrate or urea. In this experiment, a latin square design was used, in a factorial scheme (two times of incubation × two processing nylon bag). No was observed effect of the incubation time or processing in the internal markers INDF and IADF concentration and the in situ incubation after 144h is adequate to reproduce the indigestible markers fraction in samples. For fecal production estimation, the external marker chromium oxide presented similar result (1.26 kg day-1) as the total fecal collection (1.49 kg day-1). Both the internal markers overestimate the duodenal flow dry matter when compared with the external marker chromium oxide.

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.

Physical factors affecting plasmid DNA compaction in stearylamine-containing nanoemulsions intended for gene delivery

Cationic lipids have been used in the development of non-viral gene delivery systems as lipoplexes. Stearylamine, a cationic lipid that presents a primary amine group when in solution, is able to compact genetic material by electrostatic interactions. In dispersed systems such as nanoemulsions this lipid anchors on the oil/water interface confering a positive charge to them. The aim of this work was to evaluate factors that influence DNA compaction in cationic nanoemulsions containing stearylamine. The influence of the stearylamine incorporation phase (water or oil), time of complexation, and different incubation temperatures were studied. The complexation rate was assessed by electrophoresis migration on agarose gel 0.7%, and nanoemulsion and lipoplex characterization was done by Dynamic Light Scattering (DLS). The results demonstrate that the best DNA compaction process occurs after 120 min of complexation, at low temperature (4 ± 1 °C), and after incorporation of the cationic lipid into the aqueous phase. Although the zeta potential of lipoplexes was lower than the results found for basic nanoemulsions, the granulometry did not change. Moreover, it was demonstrated that lipoplexes are suitable vehicles for gene delivery. © 2012 by the authors; licensee MDPI, Basel, Switzerland.

Anti-platelet activity of infusion of Tithonia diversifolia's leaves

Tithonia diversifolia, also known as Mexican arnica, has been used in traditional medicine to treat inflammatory refractory with absence of citotoxicity. The possible health risks associated with the consumption of ingestion of the infusion (tea) plant makes it is necessary to identify the potential pharmacological activity or toxicity to prove certain plants that are acclimated in Brazil. Considering the limited number of pharmacological studies regarding the Tithonia diversifolia, the aim of this study was evaluate the effects of this infusion in platelet aggregation. Venous blood was collected with informed consent from healthy volunteers who denied taking any medication in the previous 14 days. Whole blood was transferred into polypropylene tubes containing one-tenth of final volume of acid citrate dextrose (ACD-C; citric acid 3%, trisodium citrate 4%, glucose 2%; 1:9 v/v) and centrifuged at 200g for 15 min. Platelet rich plasma was added of wash buffer solution (NaCl 140mM, KCl 5mM, sodium citrate 12mM, glucose 10mM and saccharose 12mM; pH 6; 5:7 v/v) and centrifuged at 800g for 12 min at 20°C. Platelet pellet was gently resuspended in Krebs-Ringer solution and counts were performed on a Neubauer chamber. Aggregation assay was carried out with 400 μL of platelet suspension (1.2x10 8 platelets/mL) in a cuvette at 37°C with constant stirring. Platelet suspension was incubated for 3 min with aqueous extract infusion (0.6-20μg/mL) prior to addition of thrombin (100 mU/mL). Percentage of platelet aggregation was recorded with an aggregometer (Chrono-log Lumi-Aggregometer model 560-Ca, USA). Our results show an inhibition of thrombin induced platelet aggregation in the presence of 0.6-20 ug/mL Tithonia diversifolia infusion leaves. The Tithonia diversifolia infusion leaves inhibits thrombin induced washed platelet aggregation.

Effects of ascorbic acid on in vitro culture of bovine preantral follicles

The objective of this study was to evaluate the effects of adding ascorbic acid to the media for in vitro culture of cattle ovarian fragments and to determine their effects on growth activation and viability of early-stage follicles. The ovarian cortex was divided into small fragments; one fragment was immediately fixed (control) and the other fragments were cultured in minimum essential medium (MEM) supplemented or not with various doses of ascorbic acid. Ovarian tissue was processed for histology, transmission electron microscopy (TEM) and immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA). Compared with control fragments, the percentage of primordial follicles was reduced (p < 0.05) and the percentage of growing follicles had increased (p < 0.05) in cultured cortical fragments, independent of the tested medium or incubation time. Furthermore, compared with control tissue, culture of ovarian cortex for 8 days reduced the percentages of healthy, viable follicles (p < 0.05), but not when cultures were supplemented with 25, 50 or 100 μg/ml of ascorbic acid. Ultrastructural and immunohistochemical analysis of 8 day cultured ovarian cortical fragments, however, showed the integrity and viability of follicles only when fragments were cultured in presence of 50 μg/ml of ascorbic acid. In conclusion, this study demonstrated that addition of ascorbic acid to MEM at a concentration of 50 μg/ml not only stimulates the activation of 8 day in vitro cultured cattle primordial follicles and subsequent growth of activated follicles, but also safeguards the viability of these early-stage follicles. © 2012 Copyright Cambridge University Press.

Development and Biotechnological Application of a Novel Endoxylanase Family GH10 Identified from Sugarcane Soil Metagenome

Metagenomics has been widely employed for discovery of new enzymes and pathways to conversion of lignocellulosic biomass to fuels and chemicals. In this context, the present study reports the isolation, recombinant expression, biochemical and structural characterization of a novel endoxylanase family GH10 (SCXyl) identified from sugarcane soil metagenome. The recombinant SCXyl was highly active against xylan from beechwood and showed optimal enzyme activity at pH 6,0 and 45°C. The crystal structure was solved at 2.75 Å resolution, revealing the classical (β/α)8-barrel fold with a conserved active-site pocket and an inherent flexibility of the Trp281-Arg291 loop that can adopt distinct conformational states depending on substrate binding. The capillary electrophoresis analysis of degradation products evidenced that the enzyme displays unusual capacity to degrade small xylooligosaccharides, such as xylotriose, which is consistent to the hydrophobic contacts at the +1 subsite and low-binding energies of subsites that are distant from the site of hydrolysis. The main reaction products from xylan polymers and phosphoric acid-pretreated sugarcane bagasse (PASB) were xylooligosaccharides, but, after a longer incubation time, xylobiose and xylose were also formed. Moreover, the use of SCXyl as pre-treatment step of PASB, prior to the addition of commercial cellulolytic cocktail, significantly enhanced the saccharification process. All these characteristics demonstrate the advantageous application of this enzyme in several biotechnological processes in food and feed industry and also in the enzymatic pretreatment of biomass for feedstock and ethanol production. © 2013 Alvarez et al.

Production of biofilm by Listeria monocytogenes in different materials and temperatures

Listeria monocytogenes, considered as one of the most important foodborne pathogens, is easily found on surfaces, particularly in the form of a biofilm. Biofilms are aggregates of cells that facilitate the persistence of these pathogens in food processing environments conferring resistance to the processes of cleaning and may cause contamination of food during processing, thus, representing a danger to public health. Little is known about the dynamics of the formation and regulation of biofilm production in L.monocytogenes, but several authors reported that the luxS gene may be a precursor in this process. In addition, the product of the inlA gene is responsible for facilitating the entry of the microorganism into epithelial cells that express the receptor E-cadherin, also participates in surface attachment. Thus, 32 strains of L.monocytogenes isolated from different foods (milk and vegetables) and from food processing environments were analyzed for the presence of these genes and their ability to form biofilms on three different surfaces often used in the food industry and retail (polystyrene, glass and stainless steel) at different temperatures (4, 20 and 30°C). All strains had the ilnA gene and 25 out of 32 strains (78.1%) were positive for the presence of the luxS gene, but all strains produced biofilm in at least one of the temperatures and materials tested. This suggests that genes in addition to luxS may participate in this process, but were not the decisive factors for biofilm formation. The bacteria adhered better to hydrophilic surfaces (stainless steel and glass) than to hydrophobic ones (polystyrene), since at 20°C for 24h, 30 (93.8%) and 26 (81.3%) produced biofilm in stainless steel and glass, respectively, and just 2 (6.2%) in polystyrene. The incubation time seemed to be an important factor in the process of biofilm formation, mainly at 35°C for 48h, because the results showed a decrease from 30 (93.8%) to 20 (62.5%) and from 27 (84.4%) to 12 (37.5%), on stainless steel and glass, respectively, although this was not significant (. p=0.3847). We conclude that L.monocytogenes is capable of forming biofilm on different surfaces independent of temperature, but the surface composition may be important factor for a faster development of biofilm. © 2013 Elsevier Ltd.

Produção otimizada de alginato e plástico biodegradável (poli-hidroxibutirato) por Azotobacter vinelandii

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de etanol e levana por células de Zymomonas mobilis imobilizadas em alginato

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Clarificação de suco de laranja core wash por processo de flotação auxiliado por enzimas pectinolíticas e agentes clarificantes

Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

Degradabilidade ruminal de híbridos de milho em função do estádio de colheita e processamento na ensilagem

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)