Purificação e caracterização de alfa-galactosidases de sementes de Platymiscium pubescens Micheli Purification and characterization of alpha-galactosidases from Platymiscium pubescens Micheli seeds

Este trabalho objetivou foi determinar a composição bioquímica de sementes de espécies florestais e caracterizar a enzima alfa-galactosidase de sementes germinadas de Platymiscium pubescens. Os maiores teores de lipídios foram determinados em sementes de Chorisia speciosa, Caesalpinia peltophoroides, Tabebuia serratifolia e Tabebuia velanedae, enquanto sementes de Enterolobium contortisiliquum, Schizolobium parahyba e Cassia grandis apresentaram os maiores teores protéicos. A alfa-galactosidase catalisa a hidrólise dos oligossacarídeos de rafinose, em sementes de leguminosas, durante a germinação. A maior atividade da alfa-galactosidase foi detectada em sementes de Platymiscium pubescens após 72 h de embebição. Duas formas de alfa-galactosidases, C1 e C2, foram purificadas de sementes germinadas de P. pubescens, usando-se fracionamento com sulfato de amônio e cromatografias de filtração em gel e de afinidade. Essas enzimas apresentaram atividade máxima em pH 5,5 e a 50-55 ºC. Os valores de Km ap das formas C1 e C2, para o substrato ro-nitrofenil-alfa-D-galactopiranosídeo, foram de 0,54 mM e 0,78 mM, e para a rafinose, de 4,64 mM e 5,09 mM, respectivamente. Essas enzimas exibiram estabilidade térmica moderada, mantendo 70% da atividade original após 3 h de incubação a 45 ºC. A atividade enzimática da C1 e C2 foi totalmente perdida na presença de CuSO4 e dodecil sulfato de sódio (SDS). Tais enzimas também hidrolisaram melibiose, rafinose e estaquiose, indicando potencial para aplicações biotecnológicas.<br>The objective of this work was to determine seed biochemical composition of forest species and to characterize alpha-galactosidase enzyme of germinated seeds of Platymiscium pubescens. The highest lipid levels were found in seeds of Chorisia speciosa, Caesalpinia peltophoroides, Tabebuia serratifolia and Tabebuia velanedae, whereas seeds of Enterolobium contortisiliquum, Schizolobium parahyba and Cassia grandis showed the highest protein levels. alpha-galactosidase catalyzes the hydrolyzis of raffinose oligossacarides in legume seeds during germination. The highest activity of alpha-galactosidase was found in seeds of Platymiscium pubescens after 72 h of soaking in the water. Two forms of alpha-galactosidases, C1 and C2, were purified from germinated seeds of P. pubescens, using partition with ammonium sulfate, and gel filtration and affinity chromatographies. These enzymes presented maximum activity at pH 5.5, 50-55ºC. Km ap values in the C1 and C2 forms forrho-nitrophenyl-alpha-D-galactopyranoside substrate were 0.54 mM and 0.78 mM, and 4.64 mM and 5.09 mM for raffinose, respectively. These enzymes showed moderate thermal stability, maintaining 70% of the original activity after 3 h incubation at 45ºC. The C1 and C2 enzymatic activity was totally lost in the presence of CuSO4 and sodium dodecyl sulfate (SDS). These enzymes also hydrolyzed melibiose, raffinose and stachyose, indicating a potential for biotechnological applications

Isolamento de Lactobacillusacidophilus a partir de fezes de bezerros Isolation of Lactobacillusacidophilus from calves feces

Este experimento foi realizado para isolar, caracterizar e identificar estirpes de Lactobacillusacidophilus, a partir de fezes de bezerros com 1 - 3 dias de idade, coletadas diretamente no reto. Os seguintes testes para caracterização dos isolados foram feitos: morfologia celular, teste de gram, catalase, produção de gás a partir da glicose, crescimento a 15 e 45ºC, redução do "Litmus milk", crescimento no meio com 4 e 8% de cloreto de sódio, hidrólise de arginina, redução do nitrato e fermentação de diferentes "carboidratos". De um total de 526 "isolados" iniciais, 12 (2,28%) estirpes de Lactobacillusacidophlilus foram identificadas.<br>This work was carried out to isolate, characterize and identify Lactobacillusacidophilus strain from calves feces with 1- 3 days of age, collected directly from the rectum. The following tests for the characterization of the isolates were done: cellular morphology, gran test, catalase, gas production from glucose, growth at 15 and 45ºC, Litmus milk reduction, growth in culture medium with 4 and 8% sodium chloride, arginine hydrolysis, nitrate reduction and fermentation of several "carbohydrates". From a total of 526 initial isolates, 12 (2.28%) strain of Lactobacillusacidophillus were identified

Aspectos clínicos e radiográficos do pericárdio bovino como substituto do ligamento cruzado cranial de cães Clinical and radiographic aspects of the bovine pericardium as a substitute of the canine cranial cruciate ligament

Avaliaram-se os aspectos clínicos e radiográficos do enxerto de pericárdio bovino, preservado em glicerina, como substituto do ligamento cruzado cranial. Quinze cães machos, sem raça definida, pesando entre 17,4 e 31,6 kg, foram submetidos à ruptura experimental do ligamento cruzado cranial e à substituição por pericárdio, via videoartroscopia. Os cães foram divididos em três grupos de cinco e avaliados aos 30, 90 e 120 dias. O membro operado foi imobilizado por duas semanas e procederam-se avaliações clínicas semanais. Radiografias foram feitas mensalmente e foram realizadas colheita de líquido sinovial nos tempos descritos. Clinicamente, os cães mostraram claudicação acentuada a moderada, hipotrofia muscular no membro operado e acentuado deslocamento cranial da tíbia em relação ao fêmur. As alterações degenerativas foram observadas nas radiografias. Observou-se instabilidade acentuada em todos os animais. O líquido sinovial tinha características de inflamação. Concluiu-se que o enxerto rompeu precocemente, provocou reação inflamatória persistente e fenômenos de rejeição, não sendo, portanto, recomendado para substituição do ligamento cruzado cranial de cães.<br>The clinical and radiographic aspects of the bovine pericardium preserved in glicerin, were evaluated as a substitute for canine cranial cruciate ligament. Fifteen male mongrel dogs weighing between 17.4 and 31.6kg had the ligament experimentally ruptured and the stifle joint stabilized by an arthroscopical technique with bovine pericardium as a graft. The dogs were divided into three groups of five animals each. They were evaluated at 30, 90 and 120 days. The operated limb was imobillized for two weeks and clinical examination was performed weekly. Radiographs were taken monthly and the sinovial fluid was collect at 30, 90 and 120 days. Clinically, dogs presented high to moderate lameness, muscle hipotrophy in the operated limb and accentuated cranial drawer movement. Degenerative disease was detected in radiography. All dogs showed total rupture of the graft. Sinovial fluid analysis showed characteristics of inflammation. It can be concluded that pericardium graft failed prematurely, incited persistent inflammatory reaction and rejection phenomena. Thus, it cannot be recommended as a xenograft for cranial cruciate ligament replacement

Reliability assessment using degradation models: bayesian and classical approaches

Traditionally, reliability assessment of devices has been based on (accelerated) life tests. However, for highly reliable products, little information about reliability is provided by life tests in which few or no failures are typically observed. Since most failures arise from a degradation mechanism at work for which there are characteristics that degrade over time, one alternative is monitor the device for a period of time and assess its reliability from the changes in performance (degradation) observed during that period. The goal of this article is to illustrate how degradation data can be modeled and analyzed by using "classical" and Bayesian approaches. Four methods of data analysis based on classical inference are presented. Next we show how Bayesian methods can also be used to provide a natural approach to analyzing degradation data. The approaches are applied to a real data set regarding train wheels degradation.<br>Tradicionalmente, o acesso à confiabilidade de dispositivos tem sido baseado em testes de vida (acelerados). Entretanto, para produtos altamente confiáveis, pouca informação a respeito de sua confiabilidade é fornecida por testes de vida no quais poucas ou nenhumas falhas são observadas. Uma vez que boa parte das falhas é induzida por mecanismos de degradação, uma alternativa é monitorar o dispositivo por um período de tempo e acessar sua confiabilidade através das mudanças em desempenho (degradação) observadas durante aquele período. O objetivo deste artigo é ilustrar como dados de degradação podem ser modelados e analisados utilizando-se abordagens "clássicas" e Bayesiana. Quatro métodos de análise de dados baseados em inferência clássica são apresentados. A seguir, mostramos como os métodos Bayesianos podem também ser aplicados para proporcionar uma abordagem natural à análise de dados de degradação. As abordagens são aplicadas a um banco de dados real relacionado à degradação de rodas de trens

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background: Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods: The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results: A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion: Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)