Cloning, purification and comparative characterization of two digestive lysozymes from Musca domestica larvae

cDNA coding for two digestive lysozymes (MdL1 and MdL2) of the Musca domestica housefly was cloned and sequenced. MdL2 is a novel minor lysozyme, whereas MdL1 is the major lysozyme thus far purified from M. domestica midgut. MdL1 and MdL2 were expressed as recombinant proteins in Pichia pastoris, purified and characterized. The lytic activities of MdL1 and MdL2 upon Micrococcus lysodeikticus have an acidic pH optimum (4.8) at low ionic strength (μ = 0.02), which shifts towards an even more acidic value, pH 3.8, at a high ionic strength (μ = 0.2). However, the pH optimum of their activities upon 4-methylumbelliferyl N-acetylchitotrioside (4.9) is not affected by ionic strength. These results suggest that the acidic pH optimum is an intrinsic property of MdL1 and MdL2, whereas pH optimum shifts are an effect of the ionic strength on the negatively charged bacterial wall. MdL2 affinity for bacterial cell wall is lower than that of MdL1. Differences in isoelectric point (pI) indicate that MdL2 (pI = 6.7) is less positively charged than MdL1 (pI = 7.7) at their pH optima, which suggests that electrostatic interactions might be involved in substrate binding. In agreement with that finding, MdL1 and MdL2 affinities for bacterial cell wall decrease as ionic strength increases.

Non-coding RNAs in schistosomes: an unexplored world

Non-coding RNAs (ncRNAs) were recently given much higher attention due to technical advances in sequencing which expanded the characterization of transcriptomes in different organisms. ncRNAs have different lengths (22 nt to >1, 000 nt) and mechanisms of action that essentially comprise a sophisticated gene expression regulation network. Recent publication of schistosome genomes and transcriptomes has increased the description and characterization of a large number of parasite genes. Here we review the number of predicted genes and the coverage of genomic bases in face of the public ESTs dataset available, including a critical appraisal of the evidence and characterization of ncRNAs in schistosomes. We show expression data for ncRNAs in Schistosoma mansoni. We analyze three different microarray experiment datasets: (1) adult worms' large-scale expression measurements; (2) differentially expressed S. mansoni genes regulated by a human cytokine (TNF-α) in a parasite culture; and (3) a stage-specific expression of ncRNAs. All these data point to ncRNAs involved in different biological processes and physiological responses that suggest functionality of these new players in the parasite's biology. Exploring this world is a challenge for the scientists under a new molecular perspective of host-parasite interactions and parasite development.

Development and validation of a simple and rapid capillary zone electrophoresis method for determination of nnrti nevirapine in pharmaceutical formulations

A simple and fast capillary zone electrophoresis (CZE) method has been developed and validated for quantification of a non-nucleoside reverse transcriptase inhibitor (NNRTI) nevirapine, in pharmaceuticals. The analysis was optimized using 10 mmol L-1 sodium phosphate buffer pH 2.5, +25 kV applied voltage, hydrodynamic injection 0.5 psi for 5 s and direct UV detection at 200 ��m. Diazepam (50.0 ��g mL-1) was used as internal standard. Under these conditions, nevirapine was analyzed in approximately less than 2.5 min. The analytical curve presented a coefficient of correlation of 0.9994. Limits of detection and quantification were 1.4 ��g mL-1 and 4.3 ��g mL-1, respectively. Intra- and inter-day precision expressed as relative standard deviations were 1.4% and 1.3%, respectively and the mean recovery was 100.81%. The active pharmaceutical ingredient was subjected to hydrolysis (acid, basic and neutral) and oxidative stress conditions. No interference of degradation products and tablet excipients were observed. This method showed to be rapid, simple, precise, accurate and economical for determination of nevirapine in pharmaceuticals and it is suitable for routine quality control analysis since CE offers benefits in terms of quicker method development and significantly reduced operating costs.

Oxidative DNA damage induced by S(IV) in the presence of Cu(II) and Cu(I) complexes

The DNA damage induced by S(IV) in the presence of some Cu(II) complexes in air saturated solution was investigated. The addition of S(IV) to an air saturated solution containing CuII GGA (GGA = glycylglycyl-L-alanine), CuII G3 (G3 = triglycine) or CuII G4 (G4 = tetraglycine) and Ni(II) traces, causes rapid formation of the respective Cu(III) complex, with simultaneous O2 uptake and S(IV) oxidation. SO3���- and HO��� were detected by EPR-spin trapping experiments. The DNA strand breaks were attributed to the oxysulfur radicals formed. In the reduction of Cu(II)/BCA (BCA = 4,4' dicarboxy-2-2'-biquinoline) by S(IV), with CuI BCA complex formation, there is the possible formation of carbon centered radical of BCA or peroxyl radical (ROO���) capable of oxidizing DNA bases. The intensity of DNA damage in the presence of these Cu(II) complexes and S(IV) (10-300 ��mol L-1) followed the order: CuII BCA ∼ CuII G4 ∼ Cu(II) (added as Cu(NO3)2) > CuII G3 ∼ CuII GGA. Specifically for CuII BCA the damage occurred even at lower S(IV) concentration (0.1 ��mol L-1). For the Cu(II) complexes with glycylglycylhistidine, glycylhistidylglycine, glycylhistidyllysine and glycylglycyltyrosylarginine the Cu(III) formation and the DNA damage was not observed.

Inatividade f��sica no lazer de adultos e fatores associados

OBJETIVO: Analisar a associa����o entre inatividade f��sica no lazer de adultos com fatores sociodemogr��ficos e indicadores de risco e prote����o para doen��as cr��nicas. M��TODOS: Estudo transversal com indiv��duos com idade de 18 anos e superior (n=1996). Foram utilizados dados obtidos do Sistema Municipal de Monitoramento de Fatores de Risco para Doen��as Cr��nicas N��o Transmiss��veis, por meio de entrevistas telef��nicas, em Florian��polis, SC, 2005. Analisaram-se fatores sociodemogr��ficos e comportamentais de prote����o e de risco. Os resultados das an��lises de regress��o m��ltipla para associa����o entre inatividade f��sica no lazer e vari��veis independentes foram expressos por raz��es de preval��ncia. RESULTADOS: A preval��ncia da inatividade f��sica no lazer foi de 54,6% (47,3% homens, 61,4% mulheres). Ap��s an��lise ajustada, entre os homens, maior probabilidade de inatividade f��sica no lazer foi associada ao aumento da faixa et��ria, �� diminui����o do n��vel de escolaridade e ao fato de trabalharem; menor probabilidade de inatividade f��sica no lazer foi associada ao consumo abusivo de bebida alco��lica, independentemente da faixa et��ria, n��vel de escolaridade e trabalho. Entre as mulheres, maior probabilidade de inatividade foi observada entre as que relataram n��vel de escolaridade inferior a 12 anos de estudo e que trabalhavam. An��lises ajustadas pelo n��vel de escolaridade e trabalho mostraram maior probabilidade de inatividade f��sica no lazer para mulheres que relataram consumo de frutas e hortali��as com freq����ncia inferior a cinco vezes por dia e consumo de leite integral. CONCLUS��ES: Os fatores associados �� inatividade f��sica no lazer apresentaram perfil diferente entre homens e mulheres. Para mulheres, a inatividade f��sica se associou a comportamentos de risco para doen��as cr��nicas, em especial aos h��bitos alimentares, e para os homens, se associaram a fatores sociodemogr��ficos.

��ndice de Qualidade da Dieta Revisado para popula����o brasileira

O ��ndice de Qualidade da Dieta Revisado �� um indicador de qualidade da dieta desenvolvido consoante com as recomenda����es nutricionais vigentes. Os dados diet��ticos foram provenientes do estudo de base-populacional, Inqu��rito de Sa��de e Alimenta����o (ISA)-Capital-2003. O ��ndice cont��m 12 componentes, sendo nove fundamentados nos grupos de alimentos do Guia Alimentar Brasileiro de 2006, cujas por����es di��rias s��o expressas em densidade energ��tica; dois nutrientes (s��dio e gordura saturada); e Gord_AA (calorias provenientes de gordura s��lida, ��lcool e a����car de adi����o). O ��ndice de Qualidade da Dieta Revisado propicia mensurar variados fatores de riscos diet��ticos para doen��as cr��nicas, permitindo, simultaneamente, avaliar e monitorar a dieta em n��vel individual ou populacional.

Origem e distribui����o dos nervos isqui��ticos do pre��

O pre�� do semi��rido nordestino (Galea spixii Wagler, 1831) �� um roedor pertencente �� fam��lia Caviidae. Pouca literatura �� encontrada sobre essa esp��cie em rela����o a sua morfologia e seu comportamento ambiental e reprodutivo. Com o objetivo de entender a morfologia geral, em foco, na inerva����o do membro p��lvico dessa esp��cie, neste trabalho, foi explorado o nervo isqui��tico, o qual �� o maior de todos os nervos do organismo. Foram utilizados 10 pre��s (cinco machos e cinco f��meas) que vieram a ��bito por causas naturais, oriundos do Centro de Multiplica����o de Animais Silvestres da Universidade Federal Rural do Semi��rido (CEMAS/UFERSA). Os animais foram fixados ap��s o ��bito em solu����o aquosa de formalde��do 10% e, ap��s 48 horas de imers��o nessa solu����o, foram dissecados para exposi����o do nervo isqui��tico. Dessa forma, os dados obtidos foram compilados em tabelas e expressos em desenhos esquem��ticos e fotografias. Os pares de nervos isqui��ticos originaram-se de ra��zes ventrais de L6L7S1 (70%) e de L7S1S2 (30%) e distribu��ram-se pelos m��sculos gl��teo profundo, b��ceps femural, semitendinoso e semimembranoso.

Productive performance and milk protein fraction composition of dairy cows supplemented with sodium monensin

The objective of this work was to evaluate the levels of sodium monensin on lactating cows and their effects on productive performance and milk protein fraction composition. It was used 12 Holstein cows, distributed in four balanced 3 × 3 Latin squares, and fed three diets: one control without monensin, and two diets with monensin at the levels of 24 or 48 mg/kg DM added to the concentrate. Milk production was daily measured throughout the entire experimental period. The samples used for analysis of milk composition were collected on two alternated days from the two daily milking. Non-protein nitrogen, total nitrogen and non-casein nitrogen contents were directly evaluated in the milk, and casein, whey protein and true protein contents were indirectly determined. The use of monensin in the rations reduced dry matter and nutrient intake, especially when diet with 48 mg/kg of dry matter was given. The ration with 24 mg/kg of DM increased milk production, with or without correction, and also fat and lactose yield, and it improved productive efficiency. The levels of monensin in the ratios did not influence contents of milk crude protein, non-protein nitrogen, non-casein nitrogen, true protein, casein, casein/true protein ratio, whey protein, and of all those fractions expressed as percentage of crude protein. The utilization of monensin in the ratio at the dose of 24 mg/kg of DM influences positively the productive performance of lactating cows, and it does not influence the composition of milk protein fractions.

Productive performance and composition of milk protein fraction in dairy cows supplemented with fat sources

The objective of this study was to evaluate the use of fat sources in rations for lactating cows on the productive performance and composition of milk protein fraction. Twelve Holstein cows were used, grouped in three balanced 4 × 4 Latin squares, fed with the following rations: control; refined soybean oil; whole raw soybean; and calcium salts of unsaturated fatty acid (Megalac-E). Dry matter and nutrient intake, and daily milk production were evaluated. The samples used to analyze milk composition were collected in two alternate days and were obtained from two daily milking. Milk composition and total nitrogen, non-protein nitrogen and non-casein nitrogen ratios were analyzed. The casein, serum protein and true protein ratios were obtained by difference. Dry matter and nutrient intakes were lower when cows received the diet containing calcium salts of fatty acids, in relation to the control diet. Among the diets with fat sources, the one with whole raw soybean and calcium salts decreased milk production. There was no effect of fat sources added to the diet on crude protein, non-protein nitrogen, non-casein nitrogen, true protein, casein, casein/milk true protein ratio and serum protein. Similarly, the experimental diets did not influence the protein fractions when expressed in percentage of milk crude protein. The utilization of fat sources in diets changes milk production and composition of lactating cows, but does not influence the composition of milk protein fractions.

Ornato e despojamento no mundo fabril

O artigo investiga algumas das tend��ncias da arquitetura gerada por f��bricas - galp��es industriais, moradias, igrejas, escolas, clubes etc. -, erguida no Brasil entre as duas ��ltimas d��cadas do s��culo XIX e as primeiras do XX. Apoia-se em amplo invent��rio, como base para um esfor��o de an��lise que se prop��e a identificar os temas e usos predominantes dos ornatos aplicados a constru����es geradas por f��bricas, as refer��ncias historicistas mobilizadas e eventuais rupturas de signos arquitet��nicos tipol��gicos, e, no limite, a aboli����o de ornatos e dos referidos signos. Assim, de um lado, trata da penetra����o da linguagem ecl��tica nessas constru����es, investigando o repert��rio formal utilizado em diferentes tipologias. De outro, trata da simult��nea difus��o de uma est��tica tipicamente fabril, fundamentada em no����es de economia, efici��ncia, utilidade e funcionalidade. Mostra como tais no����es se expressam ora em uma simplifica����o ou aus��ncia de ornatos, ora no uso de ornatos cujos temas remetem ao mundo das m��quinas; ��s vezes, no distanciamento ou abandono de signos arquitet��nicos tipol��gicos consagrados; ou, ainda, no emprego de materiais produzidos industrialmente e que se difundiram a partir, sobretudo, da arquitetura de f��bricas.

Acidentes de transporte em Cuiab��, Mato Grosso: morbi-mortalidade

Os acidentes de tr��nsito continuam a se constituir em um importante problema de sa��de p��blica no Brasil. Objetivo : Analisar as caracter��sticas dos acidentes de transporte terrestre e suas v��timas no munic��pio de Cuiab��. M��todo: Para o estudo da mortalidade foram obtidos dados do Sistema de Informa����es sobre Mortalidade /Minist��rio da Sa��de, dispon��veis em CD-ROM referente aos anos de 1980-2005; para o de morbidade hospitalar foram utilizados os dados Sistemas de Informa����es Hospitalares no per��odo de 1998-2006 e para o estudo da demanda das unidades de urg��ncia e emerg��ncia foi utilizado um banco de dados constru��do especialmente para esse fim, referente aos meses de maio a junho de 2005. Adotaram-se os conceitos defini����es estabelecidos na Classifica����o Internacional de Doen��as 10: acidentes de transporte (categ. V01-V99) e acidente de transporte terrestre (V01-V89). Resultados: Em todas as an��lises, as taxas de mortalidade/morbidade hospitalar se expressaram com valores maiores que a m��dia brasileira. Apesar de apresentar aspectos distintos entre mortalidade, morbidade hospitalar e morbidade da demanda de unidades de urg��ncia e emerg��ncia, destacam-se como principais v��timas os jovens do sexo masculino; a v��tima qualificada como "ocupante" predomina nos acidentes fatais e os "motociclistas", nos n��o fatais.Conclus��o: este estudo revela que Cuiab�� �� uma ��rea onde os acidentes de transporte terrestre devem ser tratados como prioridade devido �� sua magnitude, seja na mortalidade ou morbidade, trazendo subs��dios para o seu enfrentamento

Renda familiar, pre��o de alimentos e aquisi����o domiciliar de frutas e hortali��as no Brasil

OBJETIVO: Analisar a influ��ncia da renda familiar e do pre��o de alimentos sobre a participa����o de frutas e hortali��as dentre os alimentos adquiridos pelas fam��lias brasileiras. M��TODOS: Foram utilizados dados da Pesquisa de Or��amentos Familiares realizada pelo Instituto Brasileiro de Geografia e Estat��stica, com amostra probabil��stica de 48.470 domic��lios brasileiros entre 2002 e 2003. A participa����o de frutas e hortali��as no total de aquisi����es de alimentos foi expressa como percentual do total de calorias adquiridas e como calorias provenientes desses alimentos ajustadas para o total de calorias adquirido. Empregaram-se t��cnicas de an��lise de regress��o m��ltipla para estima����o de coeficientes de elasticidade, controlando-se vari��veis sociodemogr��ficas e pre��o dos demais alimentos. RESULTADOS: Observou-se aumento da participa����o de frutas e hortali��as no total de aquisi����es de alimentos com a diminui����o de seu pr��prio pre��o ou com o aumento da renda. A diminui����o do pre��o de frutas e hortali��as em 1 por cento aumentaria sua participa����o em 0,79 por cento do total cal��rico; o aumento de 1 por cento na renda familiar aumentaria essa participa����o no total cal��rico em 0,27 por cento. O efeito da renda tendeu a ser menor nos estratos de maior renda. CONCLUS��ES: A redu����o do pre��o de frutas e hortali��as, tanto pelo apoio �� cadeia de produ����o dos alimentos quanto por medidas fiscais, �� um promissor instrumento de pol��tica p��blica capaz de aumentar a participa����o desses alimentos na dieta brasileira

Effect of n-3 polyunsaturated fatty acids on oxidative stress in rats supplemented with different doses of fish oil

The diet and plasma lipid patterns associated with lipid oxidation susceptibility in rats fed different doses of polyunsaturated fatty acids (n-3 PUFA) from fish oil were evaluated. Wistar rats were assigned into three groups and received diets containing 8% soybean oil (SOY), 4% soybean oil + 4% fish oil (SOY-FISH) and 8% fish oil (FISH) for 21 days. Linoleic, oleic and ?-linolenic acids in SOY diets were substituted by myristic, palmitic, palmitoleic, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids in SOY-FISH and FISH diets reducing the n-6/n-3 ratio and increasing the peroxidability index (PI). Increased dietary EPA and DHA were observed in SOY-FISH and FISH plasma at the expense of linoleic and arachidonic acid levels. Saturated fatty acids, which were significantly different between the three diets (P < 0.01), were found at the same concentration in the plasma (P = 0.23). No changes were observed in oxidative stress as measured by the concentration of thiobarbituric acid reactive substances (TBARS) expressed in brain homogenates. However, TBARS concentration in the plasma of the SOY-FISH group was higher than the other two groups (P = 0.02). The major differences between these three groups were the n-3 PUFA content (0.4, 1.8 and 3.2 g/100 g diet) and the saturates/polyunsaturates ratio (0.3, 0.5 and 0.8) for SOY, SOY-FISH, and FISH groups, respectively. Thus, n-3 PUFA intake from fish oil only when followed by a decrease in saturated/polyunsaturated fatty acids ratio increased oxidative susceptibility in rats measured by plasma TBARS concentration

Desistindo da den��ncia ao agressor: relato de mulheres v��timas de viol��ncia dom��stica

O objetivo do estudo �� descrever a experi��ncia de mulheres, v��timas da viol��ncia dom��stica, que desistiram do processo contra seu agressor. As entrevistas foram orientadas pela quest��o: Como foi sua experi��ncia de desistir da den��ncia contra seu agressor? Das converg��ncias dos depoimentos emergiram tr��s temas: o tempo vivido da agress��o at�� a den��ncia e desist��ncia; o companheiro, a fam��lia, a delegacia da mulher; refletindo sobre a experi��ncia vivida, que descrevem o fen��meno estudado. As participantes expressam sentimentos amb��guos em rela����o ao agressor: afetividade, raiva, humilha����o e medo. Reconhecem que s��o dominadas e humilhadas. Por��m, no����es de justi��a e igualdade entre os c��njuges n��o aparecem nos depoimentos. A desist��ncia do processo pode ser compreendida na concep����o da reprodu����o da estrutura familiar, condicionada a fatores econ��micos e sociais. Os resultados remetem �� reflex��o sobre o papel da Delegacia da Mulher e das institui����es de sa��de na assist��ncia �� mulher v��tima da viol��ncia dom��stica

Searching for molecular markers in head and neck squamous cell carcinomas (HNSCC) by statistical and bioinformatic analysis of larynx-derived SAGE libraries

Background: Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignancies in humans. The average 5-year survival rate is one of the lowest among aggressive cancers, showing no significant improvement in recent years. When detected early, HNSCC has a good prognosis, but most patients present metastatic disease at the time of diagnosis, which significantly reduces survival rate. Despite extensive research, no molecular markers are currently available for diagnostic or prognostic purposes. Methods: Aiming to identify differentially-expressed genes involved in laryngeal squamous cell carcinoma (LSCC) development and progression, we generated individual Serial Analysis of Gene Expression (SAGE) libraries from a metastatic and non-metastatic larynx carcinoma, as well as from a normal larynx mucosa sample. Approximately 54,000 unique tags were sequenced in three libraries. Results: Statistical data analysis identified a subset of 1,216 differentially expressed tags between tumor and normal libraries, and 894 differentially expressed tags between metastatic and non-metastatic carcinomas. Three genes displaying differential regulation, one down-regulated (KRT31) and two up-regulated (BST2, MFAP2), as well as one with a non-significant differential expression pattern (GNA15) in our SAGE data were selected for real-time polymerase chain reaction (PCR) in a set of HNSCC samples. Consistent with our statistical analysis, quantitative PCR confirmed the upregulation of BST2 and MFAP2 and the downregulation of KRT31 when samples of HNSCC were compared to tumor-free surgical margins. As expected, GNA15 presented a non-significant differential expression pattern when tumor samples were compared to normal tissues. Conclusion: To the best of our knowledge, this is the first study reporting SAGE data in head and neck squamous cell tumors. Statistical analysis was effective in identifying differentially expressed genes reportedly involved in cancer development. The differential expression of a subset of genes was confirmed in additional larynx carcinoma samples and in carcinomas from a distinct head and neck subsite. This result suggests the existence of potential common biomarkers for prognosis and targeted-therapy development in this heterogeneous type of tumor.