Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach.

A sequential design strategy was applied to optimize the secretion of pectinases by a Saccharomyces cerevisiae strain, from Brazilian sugarcane liquor vat, on passion fruit residue flour (PFRF), through solid-state fermentation (SSF). A factorial design was performed to determine the influence variables and two rotational central composite designs were executed. The validated experimental result was of 7.1 U mL-1 using 50% PFRF (w/w), pH 5, 30 °C for 24 h, under static SSF. Polygalacturonase, pectin methyl esterase, pectin-lyase and pectate-lyase activities were 3.5; 0.08; 3.1 and 0.8 U mL-1, respectively. Shotgun proteomics analysis of the crude extract enabled the identification of two pectin-lyases, one pectate-lyase and a glucosidase. The crude enzymatic extract maintained at least 80% of its original activity at pH values and temperatures ranging from 2 to 8 and 30 to 80 °C, respectively, over 60 min incubation. Results revealed that PFRF might be a cost-effective and eco-friendly substrate to produce pectinases. Statistical optimization led to fermentation conditions wherein pectin active proteins predominated. To the extent of our knowledge, this is the first study reporting the synthesis of pectate lyase by S. cerevisiae.

PPAR Gamma: From Definition to Molecular Targets and Therapy of Lung Diseases.

Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptor superfamily that regulate the expression of genes related to lipid and glucose metabolism and inflammation. There are three members: PPARα, PPARß or PPARγ. PPARγ have several ligands. The natural agonists are omega 9, curcumin, eicosanoids and others. Among the synthetic ligands, we highlight the thiazolidinediones, clinically used as an antidiabetic. Many of these studies involve natural or synthetic products in different pathologies. The mechanisms that regulate PPARγ involve post-translational modifications, such as phosphorylation, sumoylation and ubiquitination, among others. It is known that anti-inflammatory mechanisms involve the inhibition of other transcription factors, such as nuclear factor kB(NFκB), signal transducer and activator of transcription (STAT) or activator protein 1 (AP-1), or intracellular signaling proteins such as mitogen-activated protein (MAP) kinases. PPARγ transrepresses other transcription factors and consequently inhibits gene expression of inflammatory mediators, known as biomarkers for morbidity and mortality, leading to control of the exacerbated inflammation that occurs, for instance, in lung injury/acute respiratory distress. Many studies have shown the therapeutic potentials of PPARγ on pulmonary diseases. Herein, we describe activities of the PPARγ as a modulator of inflammation, focusing on lung injury and including definition and mechanisms of regulation, biological effects and molecular targets, and its role in lung diseases caused by inflammatory stimuli, bacteria and virus, and molecular-based therapy.

Environmental and biological monitoring of benzene, toluene, ethylbenzene and xylene (BTEX) exposure in residents living near gas stations.

The volatile organic compounds benzene, toluene, ethylbenzene, and xylene (BTEX) are emitted into the atmosphere at gas stations (GS) leading to chronic exposure of nearby residents, which raises public health concerns. This study aimes at determining the contribution of GS emissions to BTEX exposure in nearby residents. Three Control and Exposed areas to BTEX emissions from GS were defined in a medium-sized European city (Porto, Portugal). BTEX atmospheric levels were determined in Control and Exposed areas using passive samplers deployed outdoors (n = 48) and indoors (n = 36), and human exposure was estimated for 119 non-smoking residents using the first urine of the day. Results showed that median BTEX outdoor and indoor concentrations were significantly higher for Exposed than Control areas, with exception of ethylbenzene and xylene indoor concentrations, where no marked differences were found. Comparison of urinary concentrations between Exposed and Control residents demonstrated no significant differences for benzene and ethylbenzene, whereas levels of toluene and xylene were significantly higher in Exposed residents. No marked correlation was obtained between atmospheric BTEX concentrations and urinary concentrations. Data indicate the potential impact on air quality of BTEX emissions from GS, which confirms the importance of these findings in urban planning in order to minimize the impact on health and well-being of surrounding populations.

Modelo ambulatorial de atendimento de nódulo de tireoide / Ambulatory care model thyroid nodule

A prevalência de detecção de nódulos na tireoide através da palpação é de aproximadamente 5%. Essa prevalência sobe para 19 a 67% quando utilizada a avaliação ecográfica. A importância da avaliação clínica dessa entidade está na necessidade de diagnosticar o câncer de tireoide que ocorre em 5 a 10% dos casos. O relato descreve o perfil dos atendimentos realizados no ambulatório de nódulos de tireoide do Hospital Municipal Dr. Mário Gatti no período de 01/05/17 a 27/07/18 de pacientes que tiveram diagnóstico ultrassonográfico de nódulo de tireoide e realizaram punção aspirativa por agulha fina, a fim de inferir a respeito da capacidade de resolutividade do ambulatório interdisciplinar (endocrinologia e cirurgia de cabeça e pescoço) no diagnóstico e tratamento do câncer de tireoide. A organização do ambulatório com atendimento integral e por equipe multidisciplinar, possibilita uma melhora na qualidade assistencial além de ser elemento facilitador para o ensino, aprendizado e pesquisa. (AU)

The rate of thyroid node detection by touch is approximately 5%. This rate goes up to a range between 19 and 67% when a sonographic evaluation is used. The importance of the clinical evaluation of those thyroid nodes is within the need of diagnosing the cancer that occurs in 5 to 10% of the cases. This paper describes the profile of consultations performed at the Outpatient Clinic Thyroid Node Unit of the Municipal Hospital Dr. Mário Gatti (HMMG) from 01/05/17 to 07/27/18, of patients who had ultrasonographic diagnostics of thyroid node and underwent fine-needle aspiration, in order to draw conclusions about the capacity of the interdisciplinary clinic (Endocrinology and Head and Neck surgery) to diagnose and treat thyroid cancer effectively. The organization of the outpatient clinic, with comprehensive care and multidisciplinary team, enables an improvement in medical assistance as well as facilitating teaching, learning and research. (AU)

Short-term evaluation of a green tea extract-based mouthrinse / Avaliação de curto prazo de um enxaguatório bucal à base de extrato de chá verde

Introduction: mechanical hygiene is still the best method forthe maintenance of oral health and prevention of biofilm formation. However, mouthrinses have been increasingly used as an adjunct support especially for post-operatory conditions or when periodontal conditions require. In this way, a number of mouthrinses are available, presenting different active ingredients with predominant antimicrobial action. Objectives: the aim of the present study was to evaluate the topical action of a green-tea extract-based mouthwash on gingival tissue. Materials and Methods: a longitudinal double-blind comparison was performed with two groups of patients clinically evaluated and diagnosed as suffering from gingivitis (n= 40). The patients were divided into two groups, according to the mouthrinse used: GT - green tea (concentration of 20mg/ml), and P - placebo, 0.9% saline solution. Clinical examination of the mucosa, tooth staining, plaque and gingival indices were peformed before and 15 days after the continuous use of the products twice a day. Results: no mucosal or tasting alterations were found, neither tooth staining regardless the mouthrinse used. Statistical assessment did not detect differences on gingival indexes between the groups before and after mouthrinses use; however, plaque indexes were significant lower in patients of GT group, compared to the P group. Conclusions: the use of phytotherapic products with active ingredients should be investigated for biofilm control, responsible for the onset and progression of periodontal disease, as well as other various oral pathologies. Considering the period of evaluation pro-posed in this study, relevant decrease in initial phase of plaque for-mation could be observed with the green tea mouthrinse. Further long-term evaluation studies should be carried-out in order to eluci-date its continuous effects on oral structures.

Introdução: higiene mecânica ainda é o melhor método para a manutenção da saúde bucal e prevenção da formação de biofilmes. No entanto, enxagues têm sido cada vez mais usado como um suporte auxiliar especialmente para as condições pós-operatórias ou quando as condições periodontais exigem. Desta forma, um número de bochechos estão disponíveis, apresentando diferentes ingredientes ativos com predominante ação antimicrobiana. Objetivos: o objetivo do presente estudo foi avaliar a ação tópica de um bochecho à base de extrato de chá verde no tecido gengival. Materiais e Métodos: a comparação longitudinal, duplo-cego, foi realizada com dois grupos de pacientes clinicamente avaliados e diagnosticados como portadores de gengivite (n = 40). Os pacientes foram divididos em dois grupos, de acordo com o enxaguatório oral usado: CV - chá verde (concentração de 20 mg / ml), e P - placebo, solução salina a 0,9%. O exame clínico da mucosa, coloração dos dentes, índice de placa e índice gengival foram realizados antes e 15 dias após o uso contínuo dos produtos duas vezes por dia. Resultados: nenhuma alteração na mucosa ou de degustação foi encontrada, assim como não foi identificado modificação na coloração do dente. A Avaliação estatística não detectou diferenças no índice gengival entre os grupos antes e depois de usar enxaguatórios. No entanto, índice de placa inferior foi menor de forma significativa nos pacientes do grupo CV, em comparação com o grupo P. Conclusões: o uso de produtos fitoterápicos com ingredientes ativos deve ser investigado para o controle de biofilme, responsável pelo aparecimento e progressão da doença periodontal. Considerando o período de avaliação proposto, a redução correspondente na fase inicial da formação de placa pôde ser observada com o enxaguatório à base de chá verde. Além disso, estudos de avaliação de longo prazo devem ser realizados, a fim de elucidar seu efeito contínuo em longo prazo sobre as estruturas orais.

Efeito da acupuntura na dor lombar: revisão de literatura / Effect of acupuncture in low back pain: Literature review

A lombalgia é o acometimento mais frequente da coluna vertebral, sendo uma das principais queixas e uma das causas mais comuns de incapacidade e morbidade. Diversos tipos de tratamento são usados no combate à dor lombar, porém, muitos pacientes não apresentam resultados com a medicina convencional. Entre os tratamentos não convencionais, a acupuntura é uma das principais alternativas para o alívio e cura dessa patologia. Com isso, o objetivo do trabalho é analisar a eficácia da acupuntura no tratamento da lombalgia, comparando-a com outros tratamentos. Para isso, realizou-se uma revisão bibliográfica com 21 estudos, randomizados, revisões de literatura e meta-análises, publicados entre os anos de 1999 e 2011, com o tema deste estudo. A acupuntura foi eficaz no tratamento da lombalgia crônica e seus resultados foram superiores aos de outros tratamentos. Sugerimos que mais estudos sejam feitos para a comprovação dos efeitos da acupuntura na lombalgia aguda.(AU)

Low back pain is the most frequent involvement of the spine, with a major complaint and one of the most common causes of disability and morbidity. Several types of treatment are used to combat back pain, but many patients do not present results with conventional medicine. Among the non-conventional treatments, acupuncture is one of the main alternatives for the relief and cure of this disease. Thus, the objective of this study is to analyze the effectiveness of acupuncture in the treatment of low back pain, comparing it with other treatments. To do so, was performed a literature review of 21 studies, randomized, literature reviews and metanalysis published between the years 1999 and 2011, with the theme of this study. Acupuncture was effective in treating chronic low back pain and its results were superior to other treatments. We suggest that further studies should be done to prove the effects of acupuncture on acute low back pain.(AU)

Further insights into chemical characterization through GC-MS and evaluation for anticancer potential of Dracaena draco leaf and fruit extracts.

The present study reports for the first time the amino acid and fatty acid compositions and the antitumoral activity of aqueous extracts obtained from Dracaena draco L. leaf and fruit. Metabolite profiles were determined by gas chromatography-ion trap-mass spectrometry (GC-IT-MS), with several amino acids, palmitic, linolenic and stearic acid being identified in the leaf extract, and only proline, oleic and stearic acid in the fruit extract. The in vitro antiproliferative activities of the extracts were tested against human colon (Caco-2), kidney (A-498), and liver (HepG2) cancer cell lines. In addition, primary cultures of normal and cancerous renal cells derived from kidney cancer patients were treated with D. draco extracts (0-400 µg/mL). Antiproliferative and cytotoxic effects were determined by the MTT assay. D. draco extracts inhibited proliferation of human colon and renal tumor cells in vitro, whereas no or weak effect was observed in HepG2 cells. Compared to the fruit extract, D. draco leaf extract exhibited stronger antiproliferative activity against all cancer cells. Our results indicate that D. draco, particularly the leaf, may be useful as a cancer chemopreventive and/or chemotherapeutic agent for colon and kidney cancers.

Comparative antihemolytic and radical scavenging activities of strawberry tree (Arbutus unedo L.) leaf and fruit.

The present study reports the antioxidant properties of Arbutus unedo L. leaf and fruit extracts using different in vitro assays including (i) reducing power, (ii) scavenging effect on DPPH free radicals, and (iii) inhibitory effect on AAPH-induced hemolysis and lipid peroxidation in human erythrocytes. All assays demonstrated antioxidant efficiency for A. unedo L. aqueous extracts, being consistently higher in the leaf. EC(50) values for reducing power and DPPH radical scavenging activities were, respectively, 0.318 ± 0.007 and 0.087 ± 0.007 mg/mL for leaf, and 2.894 ± 0.049 and 0.790 ± 0.016 mg/mL for fruit extracts. Under the oxidative action of AAPH, A. unedo leaf and fruit extracts protected the erythrocyte membrane from hemolysis (IC(50) of 0.062 ± 0.002 and 0.430 ± 0.091 mg/mL, respectively) and decreased the levels of malondialdehyde, a breakdown product of lipid peroxidation (IC(50) of 0.075 ± 0.014 and 0.732 ± 0.452 mg/mL, respectively). In accordance with antioxidant activity, phenolic content was found to be significantly higher in leaf extract. To our knowledge, this is the first time that the antioxidant activity of A. unedo species is evaluated using human biological membranes. Overall, our results suggest that A. unedo leaves are a promising source of natural antioxidants with potential application in diseases mediated by free radicals.

First report on Cydonia oblonga Miller anticancer potential: differential antiproliferative effect against human kidney and colon cancer cells.

The present study reports the phenolic profile and antiproliferative properties of quince (Cydonia oblonga Miller) leaf and fruit (pulp, peel, and seed) against human kidney and colon cancer cells. The phenolic profiles of quince methanolic extracts were determined by high-performance liquid chromatography (HPLC)/diode array detector (DAD). 5-O-Caffeoylquinic acid was always one of the two major phenolic compounds present in all extracts, except for seed. Our results revealed that quince leaf and fruit extracts exhibited distinctive antiproliferative activities. The extracts from quince leaf showed concentration-dependent growth inhibitory activity toward human colon cancer cells (IC(50) = 239.7 +/- 43.2 microg/mL), while no effect was observed in renal adenocarcinoma cells. Concerning the fruit, seed extracts exhibited no effect on colon cancer cell growth, whereas strong antiproliferative efficiency against renal cancer cells was observed for the highest concentration assayed (500 microg/mL). The antiproliferative activity of pulp and peel extracts was low or absent in the selected range of extract concentrations. This is the first report showing that C. oblonga may be useful as a cancer chemopreventive and/or chemotherapeutic agent.

Human cancer cell antiproliferative and antioxidant activities of Juglans regia L.

Several studies suggest that regular consumption of nuts, mostly walnuts, may have beneficial effects against oxidative stress mediated diseases such as cardiovascular disease and cancer. Walnuts contain several phenolic compounds which are thought to contribute to their biological properties. The present study reports the total phenolic contents and antioxidant properties of methanolic and petroleum ether extracts obtained from walnut (Juglans regia L.) seed, green husk and leaf. The total phenolic contents were determined by the Folin-Ciocalteu method and the antioxidant activities assessed by the ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. Methanolic seed extract presented the highest total phenolic content (116 mg GAE/g of extract) and DPPH scavenging activity (EC(50) of 0.143 mg/mL), followed by leaf and green husk. In petroleum ether extracts, antioxidant action was much lower or absent. Under the oxidative action of AAPH, all methanolic extracts significantly protected the erythrocyte membrane from hemolysis in a time- and concentration-dependent manner, although leaf extract inhibitory efficiency was much stronger (IC(50) of 0.060 mg/mL) than that observed for green husks and seeds (IC(50) of 0.127 and 0.121 mg/mL, respectively). Walnut methanolic extracts were also assayed for their antiproliferative effectiveness using human renal cancer cell lines A-498 and 769-P and the colon cancer cell line Caco-2. All extracts showed concentration-dependent growth inhibition toward human kidney and colon cancer cells. Concerning A-498 renal cancer cells, all extracts exhibited similar growth inhibition activity (IC(50) values between 0.226 and 0.291 mg/mL), while for both 769-P renal and Caco-2 colon cancer cells, walnut leaf extract showed a higher antiproliferative efficiency (IC(50) values of 0.352 and 0.229 mg/mL, respectively) than green husk or seed extracts. The results obtained herein strongly indicate that walnut tree constitute an excellent source of effective natural antioxidants and chemopreventive agents.

Protective effect of quince (Cydonia oblonga Miller) fruit against oxidative hemolysis of human erythrocytes.

The aim of this study was to determine the phenolic content and evaluate the antioxidant activity of quince (Cydonia oblonga) fruit. For this purpose, fruits were separated into pulps, peels and seeds and methanolic extracts were prepared. The phenolic profiles were determined by HPLC/UV and antioxidant properties were studied for their ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. The main phenolic compounds were 5-O-caffeoylquinic acid for pulp and peel (57% and 29%, respectively) and stellarin-2 for seed (18%). Total phenolics content was 2.5, 6.3 and 0.4g/kg of methanolic extract for pulp, peel and seed, respectively. Pulp and peel extracts showed similar DPPH free radical scavenging activities (EC(50) of 0.6 and 0.8 mg/ml, respectively), while seed extract presented much lower antioxidant potential (EC(50) of 12.2mg/ml). Under the oxidative action of AAPH, pulp and peel extracts showed significant protection of the erythrocyte membrane from hemolysis, in a time- and concentration-dependent manner. Seed extracts by themselves induced extensive hemolysis. These results indicate higher antioxidant activity for certain parts of quince fruit, namely pulp and peel, that may therefore represent accessible sources of natural antioxidants with potential application in nutritional/pharmaceutical fields, as preventive or therapeutic agents in diseases in which free radicals are implicated.

Evaluation of free radical-scavenging and antihemolytic activities of quince (Cydonia oblonga) leaf: a comparative study with green tea (Camellia sinensis).

This study aimed to determine the phenolic profile and to investigate the antioxidant potential of quince (Cydonia oblonga) leaf, comparing it with green tea (Camellia sinensis). For these purposes, methanolic extracts were prepared and phenolics content of quince leaf was determined by HPLC/UV. The antioxidant properties were assessed by Folin-Ciocalteu reducing capacity assay and by the ability to quench the stable free radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes.5-O-Caffeoylquinic acid was found to be the major phenolic compound in quince leaf extract. Quince leaf exhibited a significantly higher reducing power than green tea (mean value of 227.8 +/- 34.9 and 112.5 +/- 1.5 g/kg dry leaf, respectively). Quince leaf extracts showed similar DPPH radical-scavenging activities (EC50 mean value of 21.6 +/- 3.5 microg/ml) but significantly lower than that presented by green tea extract (EC50 mean value of 12.7 +/- 0.1 microg/ml). Under the oxidative action of AAPH, quince leaf methanolic extract significantly protected the erythrocyte membrane from hemolysis in a similar manner to that found for green tea (IC50 mean value of 30.7 +/- 6.7 and 24.3 +/- 9.6 microg/ml, respectively, P > 0.05). These results point that quince leaf may have application as preventive or therapeutic agent in diseases in which free radicals are involved.

Recent patents on Camellia sinensis: source of health promoting compounds.

In recent years, the scientific community, food industry, consumers and media have revealed a growing interest regarding the potential benefits of tea consumption for human health. Several studies indicate that phenolic compounds, especially flavan-3-ols (catechins), are the main components responsible for the medicinal effects of this beverage due to their recognized antioxidant properties. Antioxidants are ascribed to reduce cells and biomolecules oxidative damage caused by reactive oxygen/nitrogen species (ROS/RNS). In fact, ROS and RNS have been implicated in the oxidative deterioration of food products, as well as in the pathogenesis of several chronic and/or ageing diseases such as atherosclerosis, diabetes mellitus, chronic inflammation, neurodegenerative disorders, including Alzheimer's disease, and certain types of cancer. (-)-Epigallocatechin-3-gallate, the main component of white and green teas, for which chemical synthesis is complex, not only possesses strong antiradicalar activity but also can inhibit nitration reactions, modulate carcinogen-metabolizing enzymes, trap ultimate carcinogens and inhibit cancer cell proliferation. The recent patents on the tea manufacture, extracts preparation and health-promoting properties are reviewed in this manuscript, especially in what concerns the chemopreventive and chemotherapeutic activities of tea polyphenols.

Hypericum androsaemum infusion increases tert-butyl hydroperoxide-induced mice hepatotoxicity in vivo.

Increasing evidence regarding free radical generating agents indicates that the sustained production of high levels of reactive oxygen species (ROS) can cause hepatotoxicity. Being a short chain analog of lipid peroxide, tert-butyl hydroperoxide (t-BHP) is metabolized into free radical intermediates by cytochrome P450 in hepatocytes, which initiate lipid peroxidation, glutathione depletion and cell damage. The aim of the present study was to evaluate the putative protective effect of Hypericum androsaemum lyophilised infusion against t-BHP-induced mice hepatotoxicity in vivo, which has already been shown to be antioxidant in vitro. However, the results showed that the oral pretreatment with Hypericum androsaemum infusion (4, 20 and 100 mg/kg) for 4 days before a single intraperitoneal dose of t-BHP (1.8 mmol/kg) potentiated the t-BHP-induced hepatotoxicity. In fact, it was observed a potentiation in the depletion of total glutathione and reduced glutathione (GSH) contents and increase in oxidised glutathione (GSSG) level. Also the histopathological evaluation of the mice livers revealed that the infusion raised the incidence of liver lesions induced by t-BHP. These data do not corroborate any effect of Hypericum androsaemum infusion as hepatoprotector, but rather as a potentiator of hepatotoxicity in the present experimental conditions.

Protective activity of Hypericum androsaemum infusion against tert-butyl hydroperoxide-induced oxidative damage in isolated rat hepatocytes.

Hypericum androsaemum L. (Guttiferae) is a medicinal plant with antioxidant activity. Increasing evidence regarding free radical generating agents suggests that hepatotoxic-related disorders may involve reactive oxygen species (ROS). The purpose of this study was to investigate the protective effect of Hypericum androsaemum infusion on isolated rat hepatocytes oxidative injury induced by tert-butyl hydroperoxide (t-BHP). The results showed that pretreatment of the cells with this infusion (16, 62 and 250 microg/ml) prevented the leakage of lactate dehydrogenase (LDH) and lipid peroxidation caused by a 30-min treatment with t-BHP (1mM). However, infusion-induced alterations on glutathione homeostasis were noticed, as it was observed by the increase in glutathione oxidised form (GSSG) and depletion in total glutathione levels, which indicates that plant-derived antioxidant extracts may not be considered a generalised way of treating pro-oxidant-related diseases.