Evaluation of MMP-10 and TIMP-1 levels associated with Resveratrol supplementation

Introduction Resveratrol (RSV) is a natural phytoalexin present in abundance in grapes, berries, peanuts and red wine. The main benefits of resveratrol ingestion are due to its anti-inflammatory and antioxidant properties. This compound also induces therapeutic benefits in the remodelling of extracellular matrix (ECM), which plays an important role in the regulation of the activity of matrix metalloproteinases (MMPs) and its tissue inhibitors of metalloproteinases (TIMPs). The MMPs are a family of zinc dependent endopeptidases, with an important role in the physiological and pathological remodelling of ECM, once the activity of MMPs can be regulated by TIMPs. This regulation is essential, since if there is an imbalance between the MMPs and TIMPs there’s destruction of the ECM, leading to the development of several pathologies. Objectives Evaluate RSV potential by determining serum and salivary levels of MMP-10 and TIMP-1. Methodology The study population included 27 undergraduates between the ages of 18 and 30, divided into a control group (placebo) and an intervention group, supplemented with 100 mg RSV/day, during 30 days. MMP-10 and TIMP-1 levels were determined by slot blot. The results were analysed using a GraphPad Prism version 5 software for Windows (GraphPad Software, San Diego, California, USA). Results It was verified the existence of a linear correlation between the serum and salivary levels of TIMP-1, as well as a tendency of increase of this biomarker after RSV supplementation. In all indicators studied, supplementation with this compound wasn’t harmful. Conclusion The rising trend verified at TIMP-1 in both fluids may reflect benefits in the individuals under study, demonstrating that saliva has potential as a study biofluid for this type of biochemical markers. It was also verified that supplementation with this compound did not damage the evaluated parameters and, therefore, the RSV potential shouldn’t be rejected.

Soy Preparations Are Potentially Dangerous Factors in the Course of a Food Allergy

The special properties of soy preparations make them common additives for food production and can be dangerous for sensitive individuals. Our aim was to check consumers’ awareness of potential risks of soy preparations added to numerous food products, depending on respondents’ education, and to evaluate immunoreactive properties of chosen soy preparations. A personal questionnaire was used. Respondents (n = 251) were aged 23–28 years old, lived in Poland, and were graduates or students in their last year of food technology, medicine, and university of technology. The slot blot and Western blotting methods were used to mark immunoreactivity of soy preparations. It was shown that most respondents often or usually read labels of food products they buy. The surveyed indicated protein is the allergenic component in soy. Almost half of them were of the opinion that hydrolysis removes the allergenic properties of soy. Most of the medical students surveyed thought that people allergic to soy may consume products that contain soy preparations. The analytical results indicated that soy preparation contained protein fractions that were immunoreactive with sera of allergenic patients. It was proven that preparations, even hydrolysates, contain immunoreactive proteins that may be the source of hidden allergens, even though they are not recognized as dangerous by well-educated respondents.

Avaliação laboratorial do efeito antioxidante e anti-inflamatório do Resveratrol na função vascular

Introdução: O resveratrol é uma fitoalexina natural cujos principais benefícios devem-se às suas propriedades antiinflamatórias e antioxidantes. Os seus efeitos sugerem que este é um suplemento útil para a redução da inflamação, podendo desempenhar um papel fundamental na prevenção das doenças cardiovasculares. Este promove a vasodilataçãopela indução da síntese de óxido nítrico (NO), possui atividade antitrombótica e evita a agregação plaquetária. O fator de crescimento endotelial vascular (VEGF) é responsável pela angiogénese, sendo a sua expressão influenciada pelo resveratrol. Objetivos: Avaliar o efeito do resveratrol no perfil vascular pela determinação da pressão arterial e dos biomarcadoresinterleucina-6 (IL-6), proteína C reativa (PCR), VEGF e NO. Material e Métodos: 27 alunos com idades compreendidas entre os 18 e os 22 anos foram divididos em grupo controlo (GC) e grupo de intervenção (GI), que consumiram placebo e 100 mg de resveratrol por dia, respetivamente, durante 1 mês. A pressão arterial foi medida com aparelho automáticovalidado. Os níveis de PCR foram obtidos por imunoturbidimetria, os de IL-6 e VEGF por slot blot e a quantificação de NO por espectrofotometria. Resultados: Entre os grupos observou-se uma diminuição da pressão arterial braquial e central (??0.05). A variação negativa no NO apresentou-se superior no GC apesar das diferenças não serem significativas (p>0,05). Nos restantes marcadores avaliados não se observaram diferenças significativas. Conclusões: A ingestão regular de resveratrol parece ser uma abordagem preventiva a nível vascular, dado que modula positivamente o perfil vascular, reduzindo a pressão arterial. A variação nos níveis de NO poderá ajudar a explicar os benefícios verificados.

ANTICORPOS POLICLONAIS PARA O IMUNODIAGNÓSTICO DE VÍRUS BOVINO

Resumo O objetivo do estudo foi produzir anticorpos policlonais para vírus bovino e testar a reatividade em imunoensaios como imunofluorescência, imunoperoxidase e slot blot. Como imunógeno utilizou-se cepas e/ou isolados dos herpesvírus bovino tipo 1, 2 e 5 (BoHV-1, BoHV-2 e BoHV-5), vírus da diarreia viral bovina (BVDV), vírus respiratório sincicial bovino (BRSV), vírus da língua azul (BTV) e vírus vaccina (VACV) amplificados em cultivo celular. Os coelhos foram imunizados em intervalos regulares e o soro hiperimune produzido foi utilizado como anticorpo primário nos imunoensaios. A diluição de trabalho para cada antissoro foi determinada por diluição seriada limitante e variaram entre 1:800 e 1:51.200. As maiores diluições foram observadas para imunoperoxidase, quando comparadas com a imunofluorescência e slot blot. Diluições menores que 1:800 foram associadas com aumento de reações inespecíficas. Os antissoros anti-BoHV-1, -BoHV-5, -BVDV e -BRSV reagiram frente a isolados heterólogos em ensaios de imunofluorescência e imunoperoxidase. Conclui-se que os antissoros produzidos possuem elevadas concentrações de anticorpos específicos para os vírus e é uma alternativa para a utilização em imunoensaios.

A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin fromClostridium septicum

Clostridium septicumproduces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced byClostridium septicum.This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin ofClostridium septicumand produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.