Abstract This work aimed to compare the production of collagenolytic proteases produced by M. subtilissimus UCP1262 in submerged fermentation (SF) and solid-state fermentation (SSF) as well as extracting in aqueous two-phase system (ATPS). Collagenolytic protease production was performed in using MS-2 culture medium (SF) and soybean bran as substrate (SSF). Subsequently, the fermented liquid from both fermentations were used for the extraction of enzyme by ATPS, it was verified the influence of different variables from a factorial design 23. In SSF the highest protease and collagenolytic activities were achieved with 362.66 U/mL and 179.81 U/mL, respectively. When compared with SF (26.33 and 18.70 U/mL) higher values were obtained in the activities. The protease partitioning from SF and SSF in ATPS showed a similar profile showing higher affinity for the polymer rich phase. The highest value for the response variable purification factor (3.49) was obtained in the system using SSF. Thus, SSF shows promise as a bioprocess for extracellular production of collagenolytic proteases, using of soybean bran as substrate had used sustainable raw material, aiming application this possible enzyme in the treatment of burns and postoperative scarring.

Abstract The industrial demand for proteolytic enzymes is stimulating the search for new enzyme sources. Fungal enzymes are preferred over bacterial enzymes, and more effective and easier to extract. The aim of this work was to evaluate the potential of protease production by solid state fermentation (SSF) of Mucor subtilissimus UCP 1262, evaluate different specific activities, purify and partially characterize the enzyme in terms of biochemical as to the optimal pH and temperature. Initially, the enzyme crude extract was screened for 3 different proteolytic activities, collagenolytic (161.4 U/mL), keratinolytic (39.6 U/mL) and fibrinolytic (26.1 U/mL) in addition to conventional proteinase activity. After ammonium sulfate precipitation, the active fractions with fibrinolytic activity were dialyzed in 15 mM Tris-HCl buffer, pH 8, loaded onto DEAE-Sephadex A50 ion-exchange column and gel filtrated through Superdex 75 HR10/300. The enzyme showed a fibrinolytic maximum activity at 40 C and pH 9,0. The purified enzyme showed activity against a chromogenic chymotrypsin substrate, SDS-PAGE showing a molecular mass of approximately 70 kDa and, the specific activity of 25.93 U/mg. These characteristics suggest that the enzyme could be and efficiently produced in a simple and low-cost way using Mucor subtilissimus UCP 1262 in SSF.

Abstract Copaiba oil is a natural product used by Amazonian populations and recognized for its medicinal properties because it has significant antimicrobial activity for several pathogenic microorganisms. The present work aimed to evaluate and characterize the effect of natural oil produced by copaiba – Copaifera multijuga against multiresistant isolates of bubaline mastitis. The nitrocefin test was performed with isolates of Staphylococcus aureus from bubaline mastitis, which were 100% positive for beta-lactamase enzyme detection. Minimum Bactericidal Concentration (MBC) of 25% to 3.12% was obtained for Enterococcus faecalis and Escherichia coli and 50% and 25% for S. aureus, but Klebsiella pneumoniae and Bacillus subtilis were resistant. MBC with 12.5% and 6.25% oil were obtained for most multiresistant bubaline mastitis isolates from the states of Pernambuco, Ceará, Bahia and Alagoas. The results demonstrated the great potential of using copaiba natural oil in the treatment of buffalo mastitis.

Resumo: A mastite é uma inflamação na glândula mamária que pode acarretar perdas na produção e na qualidade do leite, gerando prejuízos econômicos para a pecuária leiteira. O tratamento é baseado na utilização de antibióticos, sendo, em muitos casos, ineficazes devido à resistência bacteriana já conhecida para esta doença. O objetivo deste trabalho foi selecionar linhagens de Streptomyces spp. produtoras de biocompostos com atividade antimicrobiana frente a isolados do gênero Staphylococcus multirresistentes de búfalas com mastite. Bem como, determinar os melhores parâmetros de produção, e avaliar a produção simultânea de ácido clavulânico. A seleção de Streptomyces spp. com capacidade de produzir compostos com atividade antimicrobiana foi realizada através da técnica bloco de gelose. Dentre as 30 espécies de Streptomyces spp. testadas, o micro-organismo Streptomyces parvulus DPUA 1573 apresentou melhores resultados, sendo capaz de inibir o crescimento de 7 isolados Staphylococcus spp. multirresistentes. Posteriormente, a espécie selecionada Streptomyces parvulus DPUA 1573 foi cultivada em diferentes condições pré-determinadas pelo planejamento fatorial 24, onde as variáveis independentes foram: concentração de soja (0,5; 1,0; 1,5%), glicose (0; 0,5; 1g/L), agitação (150; 200; 250rpm) e temperatura (28; 32; 37°C) e todos os ensaios do planejamento foram monitorados até 120 horas de cultivo. Todas as variáveis independentes influenciaram positivamente no crescimento celular, enquanto que para atividade antimicrobiana apenas as variáveis temperatura e agitação apresentaram efeitos significativos positivos. O líquido metabólito produzido por Streptomyces parvulus DPUA 1573 foi capaz de inibir o crescimento de sete Staphylococcus spp. multirresistentes. As melhores condições de cultivo para a produção de moléculas bioativas por este micro-organismo foi a 37?C, com 250rpm de agitação por período de 72 horas. Nos ensaios que apresentaram atividade antimicrobiana, foi avaliada a produção de ácido clavulânico ao longo do cultivo. A maior concentração de ácido clavulânico foi de 269,84g/L obtidas nas condições de 1,5% de farinha de soja em ausência de glicose no tempo de 96 horas. A linhagem Streptomyces parvulus DPUA 1573 foi eficiente contra Staphylococcus spp. multirresistentes isolados de mastite em búfalas, ainda apresentando concomitantemente produção de ácido clavulânico com o potencial uso farmacêutico.

Abstract: Mastitis is an inflammation in one or more mammary glands which can lead to reduction in production and quality of milk causing economic losses to dairy farming. The use of antibiotics is the key for the treatment of this disease, but in many cases ineffective due to bacterial resistance already known for this condition. The aim of this study was to select strains of Streptomyces spp. producing biomolecules with antimicrobial activity against multidrug-resistant Staphylococcus isolated from buffaloes with mastitis, as well as to determine the best production parameters to the evaluation of simultaneous production of clavulanic acid. Thirty species of Streptomyces spp. were used to selecting the greatest producer spectrum of antimicrobial activity (agar block technique), with selection of Streptomyces parvulus DPUA 1573, and 7 multidrug-resistant Staphylococcus spp. sensitive to its biocompounds. The selected strain of Streptomyces parvulus DPUA 1573 was cultured in different conditions predetermined by the factorial design 24, where the independent variables were: soybean concentration (0.5, 1.0, 1.5%), glucose (0, 0.5, 1g/L), agitation (150, 200, 250rpm) and temperature (28, 32, 37°C); all the tests were monitored up to 120 hours of cultivation. All independent variables influenced positively the cell growth, while for antimicrobial activity only the variables temperature and agitation showed positive effects. The antimicrobial bio compounds showed activity against seven multidrug-resistant Staphylococcus spp under the conditions: temperature 37°C, agitation 250rpm, with 72 hours of production process. In the tests which showed antimicrobial activity, was also assessed the production of clavulanic acid along with the cultivation. The highest concentration of clavulanic acid was 269.84g/L obtained under the conditions of 1.5% of soybean flour and absence of glucose in 96 hours. The strain Streptomyces parvulus DPUA 1573 was effective against multidrug-resistant strains of Staphylococcus spp. of mastitis from buffaloes, still showing concomitantly production of clavulanic acid for pharmaceutical use.

Abstract Modeling liquids/resins have been used to build up resin composite (RC) restorations, although there is a lack of information regarding their effects on the color stability of the latter. Therefore, the purpose of the present study was to evaluate the effects of the presence of modeling liquid between layers of RC and the finishing/polishing state of the material on color change in specimens exposed to red wine staining over time. Specimens were prepared by placing four increments (±0.5 mm thick) of RC (Filtek™ Z350 XT, 3M ESPE) into molds; half of which were prepared by applying modeling liquid (Scotchbond™ Multi-Purpose™ Adhesive, SBMP, 3M ESPE) between the layers of RC, whereas the other half were prepared without SBMP (control). Light-activation was performed after application of the final RC layer using a light-emitting diode (Radii, SDI) curing unit with an irradiance of 900 mW/cm2 for 20 s. Each group was divided according to the surface finishing protocol (n = 7): nothing (non-polished) or polishing with Sof-Lex™/diamond paste (polished). Initial colors of the specimens were evaluated with a digital spectrophotometer and the CIEL*a*b* color system. The specimens were stored in wine (37°C) for 12 months, and the color measurements were reassessed after 4, 6, and 12 months of storage. Scanning electron microscopy (SEM) analysis was performed at the end. Data were analyzed using ANOVA and Tukey’s test (α = 5%). The presence of SBMP resulted in lower overall color change of the RC as compared with the control. The non-polished specimens exhibited a significantly higher color change than the polished specimens. SEM images corroborated the previous findings. In summary, the use of modeling liquid between layers of RC shows potential for application to reduce or delay the staining process of RC over time. Moreover, polishing is essential to provide increased color stability of the RC restoration.

CONTEXTO: A tradução e a adaptação cultural são etapas que permitirão que um instrumento criado em determinado idioma e cultura possa ser usado em outro contexto cultural. O Charing Cross Venous Ulcer Questionnaire (CCVUQ) é um questionário de qualidade de vida em portadores de úlceras venosas que para ser utilizado no Brasil é necessária a execução do processo de adaptação transcultural. OBJETIVO: Traduzir e adaptar culturalmente para a população brasileira o CCVUQ. MÉTODOS: O processo consistiu em duas traduções e duas retrotraduções realizadas por tradutores independentes, da avaliação das versões, seguida da elaboração de versão consensual e de pré-teste comentado. RESULTADOS: No processo de tradução, algumas palavras foram modificadas no seu aspecto literal. A análise posterior dos resultados do pré-teste apontou necessidades de modificações de alguns termos para melhor compreensão do respondente. O tempo médio de aplicação do questionário foi de 5 minutos e 23 segundos. CONCLUSÃO: A versão na língua portuguesa do Charing Cross Venous Ulcer Questionnaire foi traduzida e adaptada para uso na população brasileira.

BACKGROUND: The translation and cultural adaptation are steps that will allow an instrument created in a particular language and culture might be used in another cultural context. The Charing Cross Venous Ulcer Questionnaire (CCVUQ) is a quality of life questionnaire in English for patients with venous ulcers that need to be translated and culturally adapted to be used in Brazil. OBJECTIVE: To translate and culturally adapt to the Brazilian the CCVUQ. METHODS: The process consisted of two translations and two back translations performed by freelance translators, evaluation of versions, followed by the development of consensus version and pre-test commented. RESULTS: In the process of translating some words and expressions were changed in its literal aspect. Pre-test evaluation indicated changes were needed for better understanding of the respondent. The average time to respond to the questionnaire was 5 minutes and 23 seconds. CONCLUSION: The Portuguese version of the Charing Cross Venous Ulcer Questionnaire was translated and adapted for use in Brazilian population.

A verificação do teor de anticorpos antitetânicos provenientes de 126 crianças de 5 a 7 anos de idade, do município de São Paulo (Brasil), revelou 41 e 21% de indivíduos susceptíveis na faixa etária de 5 a 6 anos, respectivamente. Todas as crianças de 7 anos de idade apresentaram proteção contra o tétano, revelando teor de antitoxina circulante em níveis superiores a 0,01 UI/ml. Pela análise dos resultados foi constatado que a susceptibilidade varia na razão inversa da idade. O teor médio de antitoxina tetânica encontrada variou de 0,0289 a 0,1143 UI/ml de soro, na população examinada.

Ascertainment of the antitetanic immune state in a group of 126 children, 5 to 7 years old, from the infant population of S. Paulo city (Brazil), disclosed 41% and 21% of 5 and 6-year-old individuals respectively, susceptible to tetanus. All the 7-year-old children showed protection against tetanus, revealing a circulating antitoxin content at levels superior to 0.01 IU/ml. By analysis of the results it became clear that susceptibility varies in inverse ratio to age. The mean content of tetanus antitoxin oscillated between 0.0289 and 0.1143 IU/ml of serum.

A verificação do teor de anticorpos antitetânicos provenientes de 126 crianças de 5 a 7 anos de idade, do município de São Paulo (Brasil), revelou 41 e 21% de indivíduos susceptíveis na faixa etária de 5 a 6 anos, respectivamente. Todas as crianças de 7 anos de idade apresentaram proteção contra o tétano, revelando teor de antitoxina circulante em níveis superiores a 0,01 UI/ml. Pela análise dos resultados foi constatado que a susceptibilidade varia na razão inversa da idade. O teor médio de antitoxina tetânica encontrada variou de 0,0289 a 0,1143 UI/ml de soro, na população examinada.

Ascertainment of the antitetanic immune state in a group of 126 children, 5 to 7 years old, from the infant population of S. Paulo city (Brazil), disclosed 41% and 21% of 5 and 6-year-old individuals respectively, susceptible to tetanus. All the 7-year-old children showed protection against tetanus, revealing a circulating antitoxin content at levels superior to 0.01 IU/ml. By analysis of the results it became clear that susceptibility varies in inverse ratio to age. The mean content of tetanus antitoxin oscillated between 0.0289 and 0.1143 IU/ml of serum.