Metalaxyl Degradation by Mucorales Strains Gongronella sp. and Rhizopus oryzae.

In this study, the degradation of metalaxyl was investigated in the presence of two Mucorales strains, previously isolated from soil subjected to repeated treatments with this fungicide and selected after enrichment technique. Fungal strains were characterised by a polyphasic approach using phylogenetic analysis of the Internal Transcribed Spacer (ITS) gene region, phenotypic characterisation by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) spectral analysis, and growth kinetics experiments. The strains were identified as Gongronella sp. and Rhizopus oryzae. The fungal growth kinetics in liquid cultures containing metalaxyl fits with Haldane model. Under laboratory conditions, the ability of Gongronella sp. and R. oryzae cultures to degrade metalaxyl was evaluated in liquid cultures and soil experiments. Both species were able to: (a) use metalaxyl as the main carbon and energy source; and (b) degrade metalaxyl in polluted soils, with rates around 1.0 mg kg-¹ d-¹. This suggests these strains could degrade metalaxyl in soils contaminated with this fungicide.

Drug survival and causes of discontinuation of the first anti-TNF in ankylosing spondylitis compared with rheumatoid arthritis: analysis from BIOBADABRASIL.

Treatment survival with biological therapy may be influenced by many factors, and it seems to be different among various rheumatic diseases and biological agents. The goal of the study was to compare the drug survival and the causes of discontinuation of anti-tumoral necrosis factor (anti-TNF) therapy in ankylosing spondylitis (AS) with rheumatoid arthritis (RA). Study participants were a cohort from the Brazilian Registry of Biological Therapies in Rheumatic Diseases (BIOBADABRASIL) between 2008 and 2012. The observation time was up to 4 years following the introduction of the first treatment. Gender, age, disease duration, disease activity, comorbidities, and concomitant therapies were assessed. A total of 1303 patients were included: 372 had AS and 931 had RA in which 38.7 % (n = 504) used infliximab (IFX), 34.9 % (n = 455) used adalimumab (ADA), and 26.4 % (n = 344) used etanercept (ETA). The anti-TNF drug survival of patients with AS was 63.08 months (confidence interval (CI) 60.24, 65.92) and patients with RA was 47.5 months (CI 45.65, 49.36). It was significant higher in AS (log-rank; p ≤ 0.001). Patients with RA discontinued anti-TNF more than patients with AS when adjusted to gender and corticosteroid. The adjHR (95 % CI) was 1.6 (1.14, 2.31). Female patients who were also corticosteroid users, but not of advanced age, have shown lower survival for both diseases (log-rank, p ≤ 0.001). The discontinuation rate of IFX, but not of ADA or ETA, was significantly higher in RA than in SA; HR (95 % CI) was 2.49 (1.46, 4.24). The main causes of discontinuation were ineffectiveness and adverse event in both diseases. AS patients have better drug survival adjusted to gender, age, and corticosteroid. This results appear to be related to the disease mechanism.

Antioxidant, antimicrobial and toxicological properties of Schinus molle L. essential oils.

ETHNOPHARMACOLOGICAL RELEVANCE: Schinus molle L. has been used in folk medicine as antibacterial, antiviral, topical antiseptic, antifungal, antioxidant, anti-inflammatory, anti-tumoural as well as antispasmodic and analgesic; however, there are few studies of pharmacological and toxicological properties of Schinus molle essential oils. The aim of this study was to evaluate the antioxidant and antimicrobial activities of Schinus molle leaf and fruit essential oils, correlated with their chemical composition and evaluate their acute toxicity. MATERIALS AND METHODS: The chemical composition of Schinus molle leaf and fruit essential oils were evaluated by GC-FID and GC-MS. Antioxidant properties were determined using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical and ß-carotene/linoleic acid methods. Antimicrobial properties were evaluated by the agar disc diffusion method and minimal inhibitory concentration assay. Toxicity in Artemia salina and acute toxicity with behavioural screening in mice were evaluated. RESULTS: The dominant compounds found in leaf and fruit essential oils (EOs) were monoterpene hydrocarbons, namely α-phellandrene, ß-phellandrene, ß-myrcene, limonene and α-pinene. EOs showed low scavenging antioxidant activity by the DPPH free radical method and a higher activity by the ß-carotene/linoleic acid method. Antimicrobial activity of EOs was observed for Gram+, Gram- pathogenic bacteria and food spoilage fungi. EOs showed totoxicity for Artemia salina and lower toxicity in Swiss mice. CONCLUSIONS: The result showed that EOs of leaves and fruits of Schinus molle demonstrated antioxidant and antimicrobial properties, suggesting their potential use in food or pharmaceutical industries.

Degradation of metalaxyl and folpet by filamentous fungi isolated from Portuguese (Alentejo) vineyard soils.

Degradation of xenobiotics by microbial populations is a potential method to enhance the effectiveness of ex situ or in situ bioremediation. The purpose of this study was to evaluate the impact of repeated metalaxyl and folpet treatments on soil microbial communities and to select soil fungal strains able to degrade these fungicides. Results showed enhanced degradation of metalaxyl and folpet in vineyards soils submitted to repeated treatments with these fungicides. Indeed, the greatest degradation ability was observed in vineyard soil samples submitted to greater numbers of treatments. Respiration activities, as determined in the presence of selective antibiotics in soil suspensions amended with metalaxyl and folpet, showed that the fungal population was the microbiota community most active in the degradation process. Batch cultures performed with a progressive increase of fungicide concentrations allowed the selection of five tolerant fungal strains: Penicillium sp. 1 and Penicillium sp. 2, mycelia sterila 1 and 3, and Rhizopus stolonifer. Among these strains, mycelium sterila 3 and R. stolonifer presented only in vineyard soils treated with repeated application of these fungicides and showed tolerance >1,000 mg l(-1) against commercial formulations of metalaxyl (10 %) plus folpet (40 %). Using specific methods for inducing sporulation, mycelium sterila 3 was identified as Gongronella sp. Because this fungus is rare, it was compared using csM13-polymerase chain reaction (PCR) with the two known species, Gongronella butleri and G. lacrispora. The high tolerance to metalaxyl and folpet shown by Gongronella sp. and R. stolonifer might be correlated with their degradation ability. Our results point out that selected strains have potential for the bioremediation of metalaxyl and folpet in polluted soil sites.

A new approach for determination of Na,K-ATPase activity: application to intact pancreatic ß-cells.

It has been postulated that a decrease in Na,KATPase- mediated ion gradients may be a contributing mechanism to insulin secretion. However, the precise role of the Na,K-ATPase in pancreatic ß-cell membrane depolarization and insulin secretion signalling have been difficult to evaluate, mostly because data reporting changes in enzymatic activity have been obtained in cell homogenates or membrane preparations, lacking intact intracellular signalling pathways. The aim of this work was to develop a method to characterize Na,K-ATPase activity in intact pancreatic ß-cells that will allow the investigation of putative Na,K-ATPase activity regulation by glucose and its possible role in insulin secretion signalling. This work demonstrates for the first time that it is possible to determine Na,K-ATPase activity in intact pancreatic ß-cells and that this is a suitable method for the study of the mechanisms involved in the Na,K-ATPase regulation and eventually its relevance for insulin secretion signalling.

Área viável e vascularização, pós autonomização cirúrgica, de retalho pré-fabricado por implante vascular em ratos

The aim of this study was to evaluate the effect of delay procedure on the survival and vasculature of a skin prefabricated flap with mean area rate of the 48.2cm² in a rat abdominal wall donnor site model. The skin flap was prefabricated by implantation of a distally ligated femoral pedicle into subdermal layer of the skin. Skin of the abdominal wall donnor site of 25 Wistar rats were compared in three groups of flaps receiving (A) pedicle implantation with no delay and (B) and (C) delay performed at the time of pedicle implantation. Three weeks later, the flaps in the group A and B were raised as an island flap, based on the implanted pedicle. In the group C, the implanted pedicle was severed when the flaps were raised. Seven days later, survival area in groups A, B and C were marked and the percentage survival area, with regard to whole flap area, calculated by Auto Cad R-14. The vascular density around the implanted pedicle, in groups A and B, was assessed by histological study. The mean percentage survival rate of the skin flap was 9.6 percent in the nondelayed group, 44.8 percent in the delayed group and 0.3percent in the group with implanted pedicle severed. The results of this study showed that delay procedure significantly increased (p 0.01, two-tailed Mann-Whitney test) the percentages survival of the flap and was found to have no influency over vascular density (p=0.307 two-tailed Mann-Whitney test) developed around the pedicle implantation three weeks after prefabrication.

Os objetivos do presente estudo foram avaliar os efeitos da autonomização cirúrgica sobre a extensão da área viável e da vascularização de retalhos cutâneos pré-fabricados com área média de 48,2cm² na parede abdominal de ratos. Os retalhos de pele eram pré-fabricados por implante de um pedículo femoral ligado distalmente e implantado diretamente na camada subdérmica. A pele da parede abdominal de 25 ratos Wistar foi comparada em três grupos de retalhos: grupo A, implante vascular sem autonomização e grupos B e C, implante vascular com autonomização no mesmo tempo cirúrgico. Três semanas após, os retalhos dos grupos A e B foram elevados como retalhos em ilha, nutridos pelo pedículo implantado. No grupo C, o pedículo implantado foi seccionado no momento da elevação do retalho. Sete dias após, a extensão das áreas viáveis nos grupos A, B e C foi delimitada e o percentual da área viável, em relação à área total do retalho, calculado por meio do Auto Cad R 14. A densidade vascular em torno do pedículo implantado, nos grupos A e B, foi avaliada por estudo histológico. O valor médio do percentual de área viável dos retalhos de pele foi de 9,6% no grupo onde os retalhos não foram autonomizados, 44,8% no grupo onde os retalhos foram autonomizados e 0,3% no grupo onde o pedículo implantado foi seccionado. Os resultados mostraram que o procedimento de autonomização aumentou significativamente (p 0,01, teste bicaudal de Mann - Whitney) os percentuais de área viável e não alterou estatisticamente (p = 0,307, teste bicaudal de Mann - Whitney) a densidade vascular em torno do pedículo, três semanas após o mesmo ter sido implantado.

Area viável e vascularizaçäo, pós autonomizaçäo cirúrgica, de retalhos pré-fabricado por implante vascular em ratos / Viable area and vasculature, surgical post-automation of prefabricated flap with vascular implant in rats

Os objetivos do presente estudo foram avaliar os efeitos da autonomizaçäo cirúrgica sobre a extensäo da área viável e da vascularizaçäo de retalhos cutâneos pré-fabricados com área média de 48,2cm2 na parede abdominal de ratos. Os retalhos de pele eram pré-fabricados por implante de um pedículo femoral ligado distalmente e implantado diretamente na camada subdérmica. A pele da parede abdominal de 25 ratos Wistar foi comparada em três grupos de retalhos: grupo A, implante vascular sem autonomizaçäo e grupos B e C, implante vascular com autonomizaçäo no mesmo tempo cirúrgico. Três semanas após, os retalhos dos grupos A e B foram elevados como retalhos em ilha, nutridos pelo pedículo implantado. No grupo C, o pedículo implantado foi seccionado no momento da elevaçäo do retalho. Sete dias após, a extensäo das áreas viáveis nos grupos A, B e C foi delimitada e o percentual da área viável, em relaçäo à área total do retalho, calculado por meio do Auto Cad R 14. A densidade vascular em torno do pedículo implantado, nos grupos A e B, foi avaliada por estudo histológico. O valor médio do percentual de área viável dos retalhos de pele foi de 9,6 por cento no grupo onde os retalhos näo foram autonomizados, 44,8 por cento no grupo onde os retalhos foram autonomizados e 0,3 por cento no grupo onde o pedículo implantado foi seccionado. Os resultados mostraram que o procedimento de autonomizaçäo aumentou significativamente (p< 0,01, teste bicaudal de Mann - Whitney) os percentuais de área viável e näo alterou estatisticamente (p = 0,307, teste bicaudal de Mann - Whitney) a densidade vascular em torno do pedículo, três semanas após o mesmo ter sido implantado.

Limites das ressecçöes craniofaciais oncológicas / Limits of oncological craniofacial resections

A cirurgia oncológica da base do crânio vem sendo utilizada desde a década de 1940. Um número muito grande de publicaçoes serviu para difundir o seu uso nos anos seguintes. Contudo, muito pouco se tem publicado sobre as indicaçoes desses grandes procedimentos operatórios e, principalmente, sobre as suas limitaçoes. Houve uma sensível expansao destes limites, principalmente a partir da década de 1980. Há, porém, situaçoes que exigem uma análise muito cuidadosa da indicaçao cirúrgica e podem, por vezes, contra-indicá-la. Por razoes didáticas, os limites podem ser divididos em: intracranianos, faciais e "filosóficos". Podem representar limites intracranianos: invasao de duramáter, do parênquima cerebral, da artéria carótida interna, do seio cavernoso ou dos nervos cranianos. A impossibilidade de se utilizar a microcirurgia reconstrutiva e a invasao orbitária bilateral se incluem entre os limites faciais. Finalmente, podem serdenominados limites "filosóficos" a indicaçao destes procedimentos para tumores benignos ou para lesoes malianas de tipos histológicos muito agressivos ou disseminadas. Na experiência da Disciplina de Cirurgia de Cabeça e Pescoço do Hospital das Clínicas da FMUSP, alguns fatores de pior prognostico, que permitiram delinear alguns destes limites foram: tipos histológicos de maior malignidade, tumores recidivados e necessidade de reconstruçao microcirúrgica.