Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

O Complexo Regulador em Saúde do Distrito Federal, Brasil, e o desafio da integração entre os níveis assistenciais / The Regulatory Complex for Health Care in the Federal District, Brazil and the challenge for integrating levels of health care

Resumo A integração entre os níveis de atenção é um desafio contínuo mesmo em sistemas de saúde consolidados e de alta performance. A reestruturação do sistema público de saúde do Distrito Federal, baseado no fortalecimento da Atenção Primária à Saúde (APS), na reconfiguração da atenção especializada (ambulatorial e hospitalar) e de sua rede de urgência e emergência, trouxe como desafio a necessidade de integração entre esses níveis. Fez-se necessária a criação de um dispositivo que desempenhasse o papel de "gatekeeper" conduzindo o acesso equânime, transparente e seguro para a atenção especializada e hospitalar. Neste sentido, foi estruturado o Complexo Regulador em Saúde do Distrito Federal (CRDF) e suas Centrais de Regulação (CR), orientados para a execução de um processo regulatório de acesso para os serviços de internação hospitalar, ambulatorial (procedimentos e consultas especializadas), cirurgias eletivas, alta complexidade, transporte sanitário, urgências e transplantes de pacientes do Distrito Federal e de fora dele. Este artigo descreve o processo de implantação e de implementação do CRDF e de suas CR, de forma a refletir sobre as potencialidades e desafios de seu papel enquanto instrumento de integração entre os níveis assistenciais de saúde.

Abstract The integration among levels of care is a continuous challenge even in consolidated and high performance health systems. The reform of public health system of Distrito Federal, based on the strengthening of primary health care associated to the reconfiguration of specialized attention (ambulatory and hospital) and of its of urgency and emergence network brought, as a challenge, the need for integration between these levels. Thus, became necessary to create an instrumentto perform the role of gatekeeper, leading to equanimous, transparent and safe access to specialized and hospital care. Thus, the Regulatory Complex in Health of Distrito Federal (CRDF) and its Regulatory Centers (CR) were created to carry out the regulatory process of access to care services, such as hospitalization, ambulatory care (procedures and specialized consultations), elective surgeries, complex procedures, sanitary transport, urgencies and transplants of patients of the Federal District and outside it. This article describes the process of the CRDF implementation and its CRs, aiming to reflect on the potential and challenges of its role as an instrument of integration among the levels of care.

Postmortem distribution of tetrodotoxin in tissues and body fluids of guinea pigs / 法医学杂志

OBJECTIVE@#To investigate the postmortem distribution of tetrodotoxin in tissues and body fluids of guinea pig, and to provide method and evidence for forensic identification and clinical diagnosis and treatment.@*METHODS@#Guinea pigs were intragastric administrated with 100, 50, 15 microg/kg tetrodotoxin, respectively. The poisoning symptoms were observed. The samples of heart, liver, spleen, lung, kidney, brain, stomach, intestines, bile, heart blood and urine were collected. The concentrations of tetrodotoxin in tissues and body fluids were measured with liquid chromatography-tandem mass spectrometry (LC-MS/MS).@*RESULTS@#After administrated with tetrodotoxin, all guinea pigs came out poisoning signs including tachypnea, weary and dead finally. Tetrodotoxin concentrations in lung, stomach, intestines and urine were higher, followed by blood, heart and brain. The concentration in bile was the lowest.@*CONCLUSION@#Postmortem distribution of tetrodotoxin in guinea pig is uneven. The concentration in the lung, stomach, intestines, urine and heart blood are higher, those tissues could be used for diagnosis of tetrodotoxin poisoning.

Localization of sugar residues in the stomach of three species of monkeys (Tupaiidae glis, Nycticebus cocang and Callithrix jacchus) by lectin histochemistry / Localización de los residuos de azúcar en el estómago de tres especies de monos (Tupaiidae glis, Nycticebus cocang y Callithrix jacchus) por histoquímica de lectina

The stomach of three species of non-human primates was investigated by lectin histochemistry to clarify the staining affinity and distribution patterns of their sugar residues. All gastric regions, with little differences between the deep and superficial parts of the same region, were rich in. in N-acetylglucosamine and/or neuraminic acid. Although, the superficial regions of the gastric mucosa were scanty in N-acetylgalactosamine, a- D-glucose and a -D-mannose, the deep parts of the gastric mucosa were rich in these sugars. In conclusion, there is a difference among the mucosubstances of surface and foveolar mucous cells, mucous neck cells, and gastric gland cells. This indicates heterogeneous composition of gastric mucus, or mucus molecules with variations in the degree of glycosylation of their oligosaccharide chains in the different cells which suggest that lectin binding affinity in the gastric mucosa correlated mostly to the degree of cellular differentiation.

El estómago de tres especies de primates no humanos fue investigado por histoquímica de lectinas para determinar la afinidad de tinción y los patrones de distribución de sus residuos de azúcar. Todas las regiones gástricas, con pequeñas diferencias entre las partes profundas y superficiales de la misma región, eran ricas en N-acetilglucosamina y/o ácido neuramínico. Si bien, las regiones superficiales de la mucosa gástrica eran escasas en N-acetilgalactosamina, a-D-glucosa y a-D-manosa, las partes profundas de la mucosa gástrica eran ricas en estos azúcares. En conclusión, existe una diferencia entre las mucosustancias de la superficie y células mucosas foveolares, células mucosas del cuello y células de las glándulas gástricas. Esto indica una composición heterogénea de la mucosa gástrica, o moléculas de moco con variaciones en el grado de glicosilación de sus cadenas de oligosacáridos en las diferentes células, sugieriendo que la afinidad de union de lectinas en la mucosa gástrica se relacionada principalmente con el grado de diferenciación celular.

Bromophenol concentrations in fish from Salvador, BA, Brazil

The main objective of this work is to evaluate the occurrence of bromophenols (2bromophenol, 4bromophenol, 2,4dibromophenol, 2,6dibromophenol and 2,4,6tribromophenol), in the flesh and guts in two species of the LutjanidaeFamily: Lutjanus synagris and Ocyurus chrysurus. The bromophenols were extracted by steam distillation with pentaneether (7:3 v/v), identified by reverse phase High Performance Liquid Chromatography (HPLCUV), and quantified bythe externalstandard method. Total bromophenol concentrations were similar in the muscle of both species, rangingfrom 36 ng g¹ to 349 ng g¹. The total bromophenol concentrations in stomach (ranging from 12 ng g¹ to 586 ng g¹)were slightly higher than in muscle. The presence of bromophenol in the muscles of the species under study may occuras a result of their diet. The results of this work are therefore expected to contribute toward a better understanding ofthe path of bromophenol absorption from the fish's stomach to the rest of its body.

O principal objetivo do presente trabalho foi o estudo de bromofenóis (2bromofenol, 4bromofenol, 2,4dibromofenol,2,6dibromofenol and 2,4,6tribromofenol), no músculo e estômago de duas espécies de peixes da Familia Lutjanidae: Lutjanus synagris e Ocyurus chrysurus. Os bromofenóis foramextraídos através de destilação por arraste a vapor com pentanoéter (7:3 v/v), analisados por Cromatografia Líquida de AltaEficiência e quantificados por padronização externa. As concentrações totais de bromofenóis no músculo de ambas as espécies foram similares e estiveram na faixa de 36 ng g¹ a349 ng g¹. As concentrações totais de bromofenóis no estômago (na faixa de 12 ng g¹ a 586 ng g¹) foram mais altas queno músculo. A presença de bromofenóis no músculo das espécies estudadas pode ter origem na dieta. Os resultados destetrabalho contribuirão para o melhor entendimento das rotas deabsorção de bromofenóis nos peixes.

Synchronous Adenocarcinoma and Mantle Cell Lymphoma of the Stomach

Synchronous occurrence of mantle cell lymphoma (MCL) and gastric cancer in the same patient has not yet been reported in the English literature. MCL comprises 2.5-7% of non-Hodgkin's lymphomas and is characterized by a poor prognosis with a median survival probability of 3-4 years in most series. A 62-year-old man was referred to our hospital for evaluation of an abnormal gastric lesion. The endoscopic finding was compatible with type IIc early gastric cancer (EGC) in the middle third of the stomach, and a biopsy of the lesion proved to be carcinoma. Radical total gastrectomy with splenectomy and Roux-en-Y esophagojejunostomy were performed. The resected specimen revealed two grossly separated lesions. Postoperative histological examination reported both adenocarcinoma and MCL. Immunohistochemical staining showed positivity for CD5, CD20, and cyclin D1 in the infiltrated lymphoid cells. MCL is an aggressive non-Hodgkin's lymphoma, and the current treatment approach is still unsatisfactory. Further advancements in the understanding of the synchronous occurrence of both diseases, and more efforts on investigations of treatment are needed.

c-Myc expression is related with cell proliferation and associated with poor clinical outcome in human gastric cancer

We underwent protein assay for Myc expression in 76 human gastric cancer tissues using immunohistochemistry. Expression of Myc protein was analyzed according to proliferative indices measured by flow cytometry. Levels of Myc protein expression was evaluated by correlating with biologic and clinical parameters. In 36 (47.4%) of 76 primary gastric cancers, overexpression of Myc was observed. We could observe expression of Myc protein in a significant portion of early gastric cancer (42.9%). Expression of Myc protein was demonstrated to be more frequent in poorly differentiated cancer cells (p=0.043). However, expression of Myc protein had little influence over progress or extent of the disease. Expression of Myc protein was significantly correlated with increased proliferative activity (p=0.032) and patients with high levels of Myc expression had poor disease-free survival. In a certain proportion of human gastric cancer, Myc protein may function as a regulator of cancer cell growth and expression of Myc may represent an aggressive phenotype of gastric cancer.

Functional correlation between histamine metabolism and worm expulsion in trichinella spiralis

Mucosal mast cell activity was quantified by measuring histamine forming capacity [HFC] of the gastric mucosa and histamine content in the intestinal tissues of mice infected with T. spiralis. The results were correlated with the kinetics of worm expulsion. It was found that T. spiralis resulted in significant elevation of HFC by the day 6 post infection [p.i.] which reached a maximal value at day 9, a time when approximately 50% of the established worm burden had been expelled. Histamine content of the intestinal tissues followed the same pattern. No intestinal worms were present by day 28 of infection and there was a gradual reduction in HFC and histamine content which had returned almost to control values by that time. Significant inverse correlation between individual worm burdens and HFC was detected

Tissue magnesium content and histopathological changes in non-survivors of aluminium phosphide poisoning.

The study was conducted in 30 non-survivors of Aluminium Phosphide poisoning and similar number of age and sex matched controls (fatalities as a result of road side accidents, head injury, etc). Magnesium content was estimated in brain, heart, stomach, kidney, liver and lung using atomic absorption spectrophotometer. It was found that tissue magnesium levels were not significantly different (p = NS) when comparison was carried out between controls (Group II) and Patients who were not given magnesium as part of treatment (group IB). However magnesium levels in different organs of patients who received magnesium as part of treatment were found to be significantly higher (group IA)(P < 0.01) in comparison to controls as well as patient group not treated with magnesium sulphate. Significant histopathological changes were observed in almost all the organs. The changes seem to be the result of direct tissue damage by phosphine rather than shock and anoxia which occurred in all these cases.

Evaluación de la variación temprana del pH gástrico intramural (pHi) en pacientes críticos / Valuation of gastric intramural pH early variation in critically ill patients

Se evalúan, en 80 pacientes (P) críticos, las relaciones existentes entre el pH gástrico intramural (pHi), determinando tonométricamente en la admisión y a las doce horas de evolución --considerando como valores normales a aquellos iguales o superiores a 7,35 --, y la evolución vital, así como, con la sumatoria APACHE II y los fracasos orgánicos (FO). El pHi del ingreso definió dos grupos (G) de P; el GI con tal valor normal y el G II con niveles reducidos. La mortalidad y la X ñ DS de FO fue significativamente mayor en el G en relación con el G I (p<.04 y p.05 respectivamente). En cada G se distinguió a supervivientes (S) y fallecidos (F); los S del G I mostraron una X ñ DS de pHi inicial sin diferencias con la de las 12 horas; en los F el G I pHi inicial fue similar al de los S y el de las 12 horas cayó en forma significativa (de 7,49 ñ 0,13 a 7,37 ñ 0,13; p<.01). En el G II los S ingresaron con una X ñ DS de pHi significativamente más alta que los F (p<.05) elevándose significativamente en los primeros (p<.05) y no presentándose modificaciones de valor en los F. La X ñ DS de puntos APACHE II no fue significativamente diferente entre los P de los grupos I y II. El pHi inicial normal revela, en general, un pronóstico más favorable; sin embargo, si en la evolución se reduce en más de 0,1 unidades, debe considerarse como una inadecuación de la perfusión y, por ende, como expresión de ineficacia de la autocompensación y/o del tratamiento. En el otro extremo, la elevación en más de 0,1 en P admitidos con pHi bajo (pero menor de 7,22) puede evidenciar la adecuación de la autorregulación y/o de las medidas adoptadas. Una elevación menor, inferior o nula, evidencia lo contrario. La medición mínimamente invasiva de la perfusión esplánica mediante tonometría posibilita no sólo la obtención de información pronóstica sino, y fundamentalmente, el acceso al control de la eficacia del tratamiento planteado