Pregnancy rates and luteal phase characteristics of bovine embryo recipients treated with flunixin meglumine / Taxas de prenhez e características da fase lútea de receptoras de embriões tratadas com flunixina meglumina em bovinos

The aim of this study was to evaluate the effects of flunixin meglumine administration on pregnancy rates and luteal phase characteristics in bovine embryo recipients at the moment of embryo transfer. In experiment 1, in vitro produced embryos were transferred to 184 females divided as control and treated group (recipients treated with 1.1mg/kg flunixin meglumine). In experiment 2, 22 females were divided as control group; group 2 (animals submitted to a reproductive tract manipulation similar to an embryo transfer on the 7th day after estrous); and group 3 (females submitted to a manipulation and treatment with 1.1mg/kg flunixin meglumine). In experiment 1 no difference was observed between control and treated groups (40.2% and 44.6%, respectively) for pregnancy rates. In experiment 2 no difference was observed on the length of luteal phase between groups, however, animals in group 2 presented lower plasma progesterone concentrations than the control group and group 3. Therefore, we concluded that although the administration of flunixin meglumine at the moment of embryo transfer inhibited the reduction plasma progesterone concentrations, it was not effective in increasing pregnancy rates of bovine recipients.(AU)

O objetivo deste estudo foi avaliar os efeitos da administração de flunixina meglumina sobre as taxas de prenhez e características da fase lútea da receptora no momento da transferência de embriões em bovinos. No experimento 1, embriões produzidos in vitro foram transferidos para 184 fêmeas, divididas em grupos controle e tratado (tratados com 1,1mg/kg de flunixina meglumina). No experimento 2, 22 fêmeas foram divididas em grupo controle (n=7); grupo 2 (n=8; animais submetidos à manipulação do trato reprodutivo semelhante à transferência de embriões no sétimo dia pós-cio); e grupo 3 (n=7; fêmeas submetidas à manipulação e ao tratamento com 1,1mg/kg de flunixina meglumina). No experimento 1, não foi observada diferença nos grupos controle e tratado (40,2% e 44,6%, respectivamente) para as taxas de prenhez. No experimento 2, não houve diferença na extensão da fase lútea entre os grupos, entretanto os animais do grupo 2 apresentaram concentrações plasmáticas de progesterona mais baixas que o grupo controle e o grupo 3. Portanto, conclui-se que a administração de flunixina meglumina no momento da transferência de embriões inibiu a redução das concentrações plasmáticas de progesterona, no entanto não foi eficaz para aumentar as taxas de prenhez de receptoras em bovinos.(AU)

Comportamento ingestivo de cordeiros Santa Inês alimentados com resíduo de cervejaria desidratado / Ingestive behavior of Santa Ines lambs fed rations containing dehydrated brewery residue

Objetivou-se avaliar o comportamento ingestivo de cordeiros Santa Inês alimentados com resíduo de cervejaria desidratado (RCD). Foram utilizados 35 cordeiros, machos, não castrados, com peso médio inicial de 16,00±1,69kg e, aproximadamente, 70 dias de idade. Adotou-se o delineamento inteiramente ao acaso, com cinco tratamentos e sete repetições, consistindo os tratamentos em: 0; 20; 40; 60 e 80% de inclusão de RCD na porção concentrada da ração. A inclusão de RCD na ração não influenciou o tempo de alimentação (TAL; h/dia) e a eficiência de alimentação (gFDN/h; P>0,05). O TAL obtido neste estudo apresentou valor médio de 4,90h/dia. Observou-se efeito linear decrescente (P<0,05) com a inclusão do RCD, para as eficiências de alimentação (gMS/h) e ruminação (gMS/h e gFDN/h). Contudo, efeito linear crescente (P<0,05) foi constatado para tempo de ruminação e mastigação total, bem como para o número de mastigações merícicas por dia para os animais alimentados com o subproduto. A inclusão de resíduo de cervejaria desidratado influencia o comportamento ingestivo de cordeiros Santa Inês, diminuindo a eficiência de alimentação, quando relacionada ao consumo de matéria seca por hora, e aumentando o tempo de ruminação, podendo ser adicionado em até 20% na porção concentrada da ração.(AU)

The aim of this study was to evaluate the ingestive behavior of Santa Ines lambs fed dehydrated brewer's residue (DBR). Thirty-five male lambs were used, with an initial mean weight of 16.00±1.69kg and, approximately, 70 days of age. A completely randomized design was used, with five treatments and seven replicates, the treatments being: 0; 20; 40; 60 and 80% of inclusion of DBR in the concentrated portion of the ration. The inclusion of DBR in the ration did not influence feeding time (FT; h/day) and feeding efficiency (gNDF/h; P> 0.05). The FT obtained in this study had an average value of 4.90h/day. There was a decreasing linear effect (P< 0.05) for feed efficiency (gDM/h) and rumination efficiency (gDM/h and gNDF/h). However, linear increasing effect (P< 0.05) was observed for rumination and total chewing time, as well as for the number of chews per day for animals fed with the byproduct. The inclusion of dehydrated brewer's residue in the concentrate can influence the ingestive behavior of Santa Ines lambs, reducing feed efficiency, when related to the dry matter intake per hour, and increasing the total rumination time in Santa Ines lambs, it can be added up to 20% in the concentrated portion of the ration.(AU)

Efeito de vitaminas adicionadas ao diluente ACP-104 sobre a qualidade do sêmen criopreservado de carpa comum (Cyprinus carpio) / Effect of vitamins added to the ACP-104 extender on the quality of cryopreserved semen of common carp (Cyprinus carpio)

O objetivo da presente pesquisa foi alcançado com a divisão da pesquisa em dois experimentos: (1) aperfeiçoar o protocolo de congelação utilizando água de coco em pó (ACP-104) como diluente para a criopreservação seminal de carpa comum; (2) avaliar o efeito da suplementação das vitaminas C (ácido ascórbico) ou E (α-tocoferol) sobre os melhores diluidores testados no experimento 1 na qualidade do sêmen pós-descongelado da espécie. Para o experimento 1, foram formados oito pools de sêmen, provenientes de 14 machos selecionados. As amostras seminais coletadas foram avaliadas quanto à motilidade total, à velocidade, ao percentual de espermatozoides normais e à vitalidade espermática antes e depois da criopreservação seminal. Esta foi realizada em meio ACP-104 acrescido de dimetilsulfóxido (DMSO), ou etilenoglicol (EG), ou glicerol, ou metanol, todos à concentração de 10%, diluídos em 1:3 (sêmen:diluidor). As amostras foram, então, congeladas em vapor de nitrogênio líquido em dry shipper e estocadas em nitrogênio líquido (-196°C). Para o experimento 2, foram formados oito pools provenientes da coleta de sêmen de 15 machos. As amostras seminais foram avaliadas seguindo as mesmas análises do experimento 1, acrescentando-se a duração da motilidade total. A criopreservação seminal utilizou-se do meio ACP-104 acrescido de DMSO ou EG, suplementado ou não com vitamina C ou E. Os melhores resultados encontrados no experimento 1 foram obtidos com o DMSO e o EG. Estes não diferiram significativamente entre si para a motilidade total (24% e 28%; P>0,05) e a normalidade espermática (32% e 26%; P>0,05), respectivamente. Para o experimento 2, o EG suplementado com vitamina E produziu significativamente resultados superiores de motilidade total, normalidade espermática e duração da motilidade em relação ao DMSO, concluindo-se que o EG deve ser, portanto, o crioprotetor de escolha a ser utilizado com o ACP-104 suplementado ou não com vitamina E.(AU)

The objective was achieved by dividing the research into two experiments: (1) improving the freezing protocol using powdered coconut water (ACP-104) as a diluent for the cryopreservation seminal of common carp; (2) evaluating the effect of supplementation of vitamins C (ascorbic acid) or vitamin E (α-tocoferol) with the best extenders tested in experiment 1 on the quality of post-thawed. For experiment 1, semen pools from 14 selected males were formed. Seminal samples were evaluated for total motility, velocity, percentage of normal sperm and sperm vitality before and after the seminal cryopreservation. This was done in ACP-104 extender plus dimethyl sulfoxide (DMSO), or ethylene glycol (EG), or glycerol or methanol all at concentration 10% diluted in 1:3 (semen:extender). The samples were frozen in vapors of nitrogen into dry shippers and stored in liquid nitrogen (-196 °C). For experiment 2, eight pools were formed from the 15 males. The semen samples were evaluated following the same analysis of experiment 1 adding duration of total motility. The sperm cryopreservation was performed in extenders ACP-104 plus DMSO or EG supplemented or not with vitamin C or E. The best results found in Experiment 1 were obtained with DMSO and EG. They do not differ significantly for total motility (24% and 28%; P>0.05) and normal sperm (32% and 26%; P>0.05) respectively. For experiment 2, EG supplemented with vitamin E, produced significantly better results overall motility, sperm normality and duration of motility relative to DMSO. In conclusion, EG should be the cryoprotectant of choice for use with the ACP-104 supplemented or not with vitamin E.(AU)

Sorovares e perfil de suscetibilidade a antimicrobianos em Salmonella spp. isoladas de produtos de origem suína / Serovars and antimicrobial susceptibility of Salmonella spp. product isolated from swine

Foi analisado um total de 1824 cepas de Salmonella, isoladas de alimentos de origem suína, no período de janeiro/2005 a junho/2010. As cepas, provenientes de diferentes regiões do país, foram recebidas pelo Labent/IOC/FIocruz para caracterização antigênica conclusiva. Foram identificados 41 sorovares, destacando-se: Typhimurium, Derby, Enteritidis, Panama, Infantis e Anatum. Aspectos bacteriológicos e epidemiológicos relacionados a esses sorovares foram discutidos. O teste de suscetibilidade aos antimicrobianos foi realizado em 357 amostras, 257 (72%) foram resistentes a uma ou mais drogas, e destas, 31,9% mostraram-se multirresistentes. A variedade de sorovares observada neste estudo confirma o papel dos suínos na cadeia alimentar como importantes reservatórios de Salmonella, agravado ainda pelo elevado percentual de cepas resistentes a um ou mais antimicrobianos, alertando para uma condição de risco à saúde pública.

We analyzed a total of 1824 strains of Salmonella isolated from swine-origin foods from January/2005 to June/2010. The strains from different regions of the country were received by Labent/IOC/FIOCRUZ for conclusive antigenic characterization. We identified 41 serovars, of which these stood out: Typhimurium, Derby, Enteritidis, Panama, Infantis and Anatum. Bacteriological and epidemiological aspects related to these serovars were discussed. The antimicrobial susceptibility test was performed on 357 samples, 257 (72%) were resistant to one or more of these drugs and 31,9% were multiresistant. A variety of serovars were identified reinforcing the swine as an important reservoir of Salmonella in the food chain. The high rates of antimicrobial resistance obtained in this evaluation may represent a risk condition to human health.

Níveis de treonina digestível para codorna japonesa na fase de produção / Digestible threonine levels for laying japanese quail

Avaliou-se a exigência de treonina digestível para a codorna japonesa em postura. Foram utilizadas 400 codornas, durante 63 dias. Utilizou-se delineamento inteiramente ao acaso, com cinco tratamentos, oito repetições e 10 aves por unidade experimental. A dieta basal foi deficiente em treonina, contendo 17,8% de proteína bruta, 2.900kcal de EM/kg, sendo suplementada com quatro níveis de L- treonina 98% (0,000; 0,052; 0,104; 0,156 e 0,208%), correspondendo à relação treonina digestível:lisina digestível de 0,55; 0,60; 0,65; 0,70 e 0,75, respectivamente. Os parâmetros estudados foram: consumo de ração, consumo de treonina, produção de ovo, peso do ovo, massa de ovos, conversão alimentar por massa de ovos, conversão alimentar por dúzia de ovos, componentes dos ovos (gema, albúmen e casca) e gravidade específica. Observou-se aumento linear (P<0,01) somente para o consumo de treonina. Conclui-se que, para proporcionar os melhores resultados de desempenho e qualidade de ovos, a codorna japonesa exige 0,55% de treonina digestível, para um consumo diário de 144,61mg de treonina digestível/ave, correspondendo à relação treonina digestível: lisina digestível de 0,55.

This experiment was conducted to determine the digestible threonine requirement for laying Japanese quail. Four hundred quails were used during 63 days. A completely randomized experimental design was used with five treatments, eight replicates and ten quails per experimental unit. The basal diet was deficient in threonine, containing 17.8% crude protein, 2.900kcal of ME/kg and supplied with four levels of L-threonine 98% (0.000; 0.052; 0.104; 0.156 and 0.208%), corresponding digestible threonine: digestible lysine ratio 0.55; 0.60; 0.65; 0.70 and 0.75, respectively. The following parameters were evaluated: feed intake, threonine intake, egg production, egg weight, egg mass, feed conversion per egg mass, feed conversion per egg dozen, egg quality (yolk, albumen and shell) and specific gravity. Linear increase (P<0.01) was only observed in the threonine intake. The conclusion is that the Japanese quail demand 0.55% of digestible threonine to provide the best productive performance and quality of eggs, a daily consumption of 144.61 mg of digestible threonine/quail, corresponding to the digestible threonine: digestible lysine ratio of 0.55.

Gluttonous predators: how to estimate prey size when there are too many prey / Predadores glutões: como estimar o tamanho das presas quando o número delas é muito grande

Prey size is an important factor in food consumption. In studies of feeding ecology, prey items are usually measured individually using calipers or ocular micrometers. Among amphibians and reptiles, there are species that feed on large numbers of small prey items (e.g. ants, termites). This high intake makes it difficult to estimate prey size consumed by these animals. We addressed this problem by developing and evaluating a procedure for subsampling the stomach contents of such predators in order to estimate prey size. Specifically, we developed a protocol based on a bootstrap procedure to obtain a subsample with a precision error of at the most 5 percent, with a confidence level of at least 95 percent. This guideline should reduce the sampling effort and facilitate future studies on the feeding habits of amphibians and reptiles, and also provide a means of obtaining precise estimates of prey size.

O tamanho das presas é uma importante dimensão do nicho trófico. Em estudos de ecologia alimentar, os itens alimentares são geralmente medidos individualmente com o uso de paquímetro ou ocular micrométrica. Entre os anfíbios e répteis, há espécies que consomem grande número de itens alimentares pequenos (e.g. formigas, cupins). Esse grande número, por sua vez, torna a estimativa do tamanho das presas consumidas uma tarefa difícil. Desenvolvemos um método para colher subamostras dos conteúdos estomacais desses animais com o objetivo de obter estimativas de tamanho das presas. Especificamente, desenvolvemos um protocolo baseado em uma rotina de bootstrap que permite a obtenção de subamostras com erro de precisão de no máximo 5 por cento e confiança de 95 por cento. Esse método deve diminuir o esforço amostral e facilitar estudos futuros sobre os hábitos alimentares de anfíbios e répteis, além de fornecer um meio de obter estimativas precisas de tamanho de presas.

The defensive strike of five species of lanceheads of the genus Bothrops (Viperidae)

We studied the defensive strike of one species of each of five recognized lineages within the genus Bothrops, namely, B. alternatus, B. jararaca, B. jararacussu, B. moojeni and B. pauloensis. The defensive strike of the studied species was in general similar to that of Crotalus viridis and C. atrox, but some important differences were observed. Bothrops alternatus and B. pauloensis struck preferentially from a tight body posture, whereas B. jararaca and B. moojeni from a loose body posture. Defensive strikes were either true or false (during the latter, the mouth remains closed or partially open). Almost all strikes were successful; only on a few occasions snakes missed their target (flawed strikes). Strike variables were very conservative among the five species, especially strike distance and height, and one possible explanation may be related to constraints imposed on strike variables as a way of increasing strike accuracy.

Estudamos o bote defensivo de uma espécie de cada uma de cinco reconhecidas linhagens do gênero Bothrops, a saber: B. alternatus, B. jararaca, B. jararacussu, B. moojeni e B. pauloensis. O bote defensivo das espécies estudadas foi, em geral, semelhante ao de Crotalus viridis e C. atrox, porém algumas diferenças foram observadas. Bothrops alternatus e B. pauloensis desferiram botes preferencialmente a partir de postura corpórea enrodilhada, ao passo que B. jararaca e B. moojeni desferiram a maioria dos botes a partir de postura corpórea frouxa. Os botes defensivos foram verdadeiros ou falsos (nestes, a boca da serpente permaneceu fechada ou parcialmente aberta). Quase todos os botes foram bem-sucedidos; apenas em alguns casos a serpente errou o alvo (botes falhos). As variáveis relativas aos botes foram bastante conservativas entre as cinco espécies, principalmente distância e altura do bote, e uma possível explicação pode estar relacionada a restrições impostas às variáveis relativas aos botes como forma de aumentar sua acurácia.

Negative inotropic and chronotropic effects on the guinea pig atrium of extracts obtained from Averrhoa carambola L. leaves

It has been reported that star fruit can lead to a fatal outcome in uremic patients. The intoxication syndrome consists of hiccups, mental confusion, dizziness, and vomiting. On the other hand, folk medicine uses teas and infusions of carambola leaves to treat headache, vomiting, cough, insomnia, and diabetes. This motivated us to determine if Averrhoa carambola can act on the contractility and automaticity of the guinea pig heart. We measured the atrial isometric force in stimulated left atria and determined the chronotropic changes in spontaneously beating right atria. The carambola leaf extracts (1.5 mg/ml) abolished the contractile force in a concentration-dependent manner. Among the crude, methanolic, ethanolic, aqueous, and acetic extracts, the aqueous one was the most potent (EC50 = 520 ± 94 æg/ml; flavonoids and tannins are the main constituents; Na+ and K+ contents in 1.0 mg/ml of aqueous extract were 0.12 ± 0.016 and 1.19 ± 0.15 mM, respectively). The aqueous extract abolished the positive Bowditch staircase phenomenon and reduced the inotropic response to CaCl2 (0.17-8.22 mM), events that are dependent on the cellular Ca2+ inward current. The adrenergic, muscarinic or opioid membrane receptors do not seem to participate in the mechanism of action of the cardioactive substance(s). In spontaneously beating atria, the aqueous extract promoted a negative chronotropic effect that was antagonized by 0.1 æM isoproterenol bitartrate. With this agonist, the EC50 of the aqueous extract increased from 133 ± 58 to 650 ± 100 æg/ml. These data regarding the effect of A. carambola on guinea pig atrial contractility and automaticity indicate an L-type Ca2+ channel blockade.

Antioxidant dietary deficiency induces caspase activation in chick skeletal muscle cells

Apoptosis and necrosis are two distinct forms of cell death that can occur in response to different agents and stress conditions. In order to verify if the oxidative stress induced by dietary selenium and vitamin E deficiencies can lead muscle cells to apoptosis, one-day-old chicks were reared using diets differing in their vitamin E (0 or 10 IU/kg) and selenium (0 or 0.15 ppm) supplementation. Chick skeletal muscle tissue was obtained from 28-day-old animals and used to verify apoptosis occurrence based on caspase activity detection and DNA fragmentation. Antioxidant deficiency significantly increased caspase-like activity assessed by the hydrolysis of fluorogenic peptide substrates (Abz-peptidyl-EDDnp) at lambdaexc = 320 nm and lambdaem = 420 nm. Proteolytic activation was not accompanied by typical internucleosomal DNA fragmentation detected by field inversion gel electrophoresis. Although the general caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone (Z-VAD-fmk) (0 to 80 muM) did not block caspase-like activity when preincubated for 30 min with muscle homogenates, the hydrolyzed substrates presented the same cleavage profile in HPLC (at the aspartic acid residue) when incubated with the purified recombinant enzyme caspase-3. These data indicate that oxidative stress causes caspase-like activation in muscle cells and suggest that cell death associated with exudative diathesis (dietary deficiency of selenium and vitamin E) can follow the apoptotic pathway

Glycosaminoglycans affect the interaction of human plasma kallikrein with plasminogen, factor XII and inhibitors

Human plasma kallikrein, a serine proteinase, plays a key role in intrinsic blood clotting, in the kallikrein-kinin system, and in fibrinolysis. The proteolytic enzymes involved in these processes are usually controlled by specific inhibitors and may be influenced by several factors including glycosaminoglycans, as recently demonstrated by our group. The aim of the present study was to investigate the effect of glycosaminoglycans (30 to 250 æg/ml) on kallikrein activity on plasminogen and factor XII and on the inhibition of kallikrein by the plasma proteins C1-inhibitor and antithrombin. Almost all available glycosaminoglycans (heparin, heparan sulfate, bovine and tuna dermatan sulfate, chondroitin 4- and 6-sulfates) reduced (1.2 to 3.0 times) the catalytic efficiency of kallikrein (in a nanomolar range) on the hydrolysis of plasminogen (0.3 to 1.8 æM) and increased (1.9 to 7.7 times) the enzyme efficiency in factor XII (0.1 to 10 æM) activation. On the other hand, heparin, heparan sulfate, and bovine and tuna dermatan sulfate improved (1.2 to 3.4 times) kallikrein inhibition by antithrombin (1.4 æM), while chondroitin 4- and 6-sulfates reduced it (1.3 times). Heparin and heparan sulfate increased (1.4 times) the enzyme inhibition by the C1-inhibitor (150 nM)

Comparacao de dois protocolos de fortalecimento para preensao plamar / Comparison of two protocols for increasing grip strength

A medida da forca de preensao palmar tem sido um dos meios uteis para a avaliacao das caracteristicas fisicas, a evolucao durante a reabilitacao e o grau de incapacidade. Na pratica clinica o fisioterapeuta se depara com a dificuldade em selecionar o protocolo mais adequado e eficiente para o fortalecimento muscular. O objetivo deste trabalho foi comparar dois protocolos de regimes isotonicos buscando evidenciar qual deles seria o mais indicado para o fortalecimento musculos responsaveis pela preensao. As medidas da forca de preensao pre e pos-treinamento foram realizadas com um dinamometro mecanico da marca Jamar, em 22 mulheres com idade entre 18 e 21 anos (media de 19 anos). As voluntarias foram separadas em 2 grupos, DeLorme (carga crescente) e Oxford (carga decrescente), para a aplicacao dos protocolos. Para o treinamento foi utilizado um exercitador de mao e dedos da marca Digi-flex, e o programa foi realizado duas vezes por semana, com duracao de 20 minutos cada sessao em um total de 8 sessoes. Posteriormente, foram comparadas as forcas pre e pos-treinamentos e foi observado que, para as amostras dependentes (pre e pos-treinamentos), houve diferenca significativa tanto no grupo DeLorme quanto no Oxford, ja para as amostras independentes (pos-DeLorme e pos-Oxford) nao houve diferenca significativa

Variaçäo temporal de características clínicas em bovinos mestiços alimentados com polpa cítrica peletizada / Clinical characteristics in crossbred cattle fed citrus pulp pellets

Cinco bovinos mestiços sem raça definida, entre 24 e 36 meses de idade e média de peso de 150kg, foram submetidos a alimentaçäo com dieta completa, na qual a polpa cítrica peletizada comercial correspondia a 40 por cento da ingestäo de matéria seca diária durante 30 dias, com o objetivo de avaliar possíveis alteraçöes na temperatura retal, freqüências respiratória e cardíaca, tempo de preenchimento capilar e freqüência de movimentos ruminais, e no peso corporal. Os animais apresentaram aumento na freqüência de movimentos ruminais e ganharam em média 860g de peso/dia durante o experimento. Todas as variáveis clínicas estavam dentro do intervalo de normalidade para a espécie

Isolation and partial characterization of an endopeptidase from Enterolobium contortisiliquum seeds

1. Aqueous extracts of Enterolobium contortisiliquum seeds contain an endopeptidase of M(r) 60,000 with specificity for basic amino acid residues. The enzyme was purified by chromatography on DEAE Sephadex, followed by gel filtration on Sephadex G-75 and affinity chromatography on Zinc-Sepharose. The overall purification was 300-fold and the yield about 46. 2. The endopeptidase hydrolyzes benzoyl-arginine-p-nitroanilide (Bz-Arg-pNan) and acetyl-phenylalanine-arginine-p-nitroanilide (Ac-Phe-Arg-pNan) with Km 14.4 mM and 0.062 mM, respectively. Succinyl-phenylalanine-p-nitroanilide (Suc-Phe-pNan) and tosyl-arginine methyl ester (TAME) were not hydrolyzed. E. contortisiliquum endopeptidase also cleaves a seed protein of low molecular weight from the same E. contortisiliquum seeds, and converts Met-Lys-bradykinin into bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg). 3. Metals (1.0 mM) such as Cr3+, Fe3+ and Zn2+ ions inactivate the enzyme when Bz-Arg-pNan was the substrate. Enzyme activity is abolished by EDTA but is partially restored by Cu2+, Al3+, Ba2+, Mn2+, Mg2+, Fe2+, Ca2+ and Co2+ ions. The endopeptidase is not inhibited by the previously purified E. contortisiliquum inhibitors of trypsin and cysteine proteinases, or by soybean trypsin inhibitor (Oliva et al. (1987). Brazilian Journal of Medical and Biological Research, 20:767-770).

Purification and characterization of endopeptidase H2, a kinin inactivating serine proteinase (kininase) from human urine

1. A kinin-inactivating chymotrypsin-like serine-endopeptidase was purified 202-fold from human urine by DEAE-cellulose chromatography, gel filtration, DEAE/HPLC chromatography and affinity chromatography. It hydrolyzed bradykinin at the Phe5-Ser6 peptide bond at a rate of 1.090 mumol min-1 mg protein-1 at pH 8.0 and 37 degrees C. The molecular weight of this endopeptidase H2, estimated by SDS-polyacrylamide gel electrophoresis and by gel filtration, was 60 kDa, and its optimum pH for bradykinin hydrolysis was near 8.5. 2. Bradykinin inactivating activity was inhibited 100 per cent by the serine-proteinase inhibitor PMFS (1 mM) and the chymotrypsin inhibitor TPCK (5 mM). Reagents such as 2-mercaptoethanol (3 mM) and pOH-mercuribenzoate (3 mM) inhibited the enzyme by 100 per cent and 67 per cent , respectively. 3. Endopeptidase H2 hydrolyzes the Phe-Ser bond of peptides related to bradykinin and its activity appears to be limited to peptide chains of < or = 18 amino acid residues since it does not hydrolyze BAM 22, peptide E or kininogen. 4. The molecular size and inhibition profile suggested that endopeptidase H2 differs from the serine-proteinases previously described in rat liver, rat hepatic endothelium, rat and rabbit brain. 5. The physiological role of endopeptidase H2 may be a link between the kinin and neuropeptide systems in the control of water-electrolyte balance

Endopeptidase and carboxypetidase activities in human urine which hydrolyze bradykinin

We have fractionated the bradykinin inactivating activity of human urine by stepwise elution chromatography on DEAE-cellulose and recovered 95% of the inactivating activity and 29% ofd the protein (absorbance at A280 nm). Seven of nine fractions which presented activity were also tested for angiotensin I and II inactivating activity, angiotensin convertingg activity and for the hydrolysis of hippuryl-His-Leu and hippuryl-Arg. Sites of hydrolysis in bradyykinin were determined by HPLC of the hydrolysates and fragments were compared with authentic peptides. Cleavage sites demonstrated for Fractioons A through G were: Phe8-Arg9 (A and B), Phe5-Ser6 (C and F), Pho7-Phe8 (D), Gly4-Phe5 and Pro7-Phe8 (E) and Pro3-Gly4 (G). The relative molecular weight of the bradykininase activity present in each fraction, determined by gel filtration, was: 16 kDa (A), 70 kDa (B), 60 kDa (B) (C), 88 kDa (D), 230 kDa (E) and 49 kDa (G). Bradykinin inactiivating activity was inhibited 50--100% by 3 mM EDTA (A,B,D,E adn G), 1 mMM 2-mercaptoethanol (A,B,C and G), 0.1 mM PMSF (C and F), 1 mM TPCK (C and F), 1 mM Xn2+ (C), 60 uM BPP5a and 40 uM BPP9a (D), 0.1 uM phosphoramidon (E) and 3 mM sodium p-hydroxyymercuribenzoate (G). The properties of some of these bradykinin inactivating activities correspondend to enzymes previously described in urine and tissues: carboxypeptidases (Fractions A and B, angiotensin I converting enzyme (Fraction d), neutral endopeptidase (Fraction E). However, the chymotrypsin-like activity of fractions C and F and the prolylendopeptidase activity of fraction G have not been described before in urine and they being purified in order to obtain a more accurate characterization

Estudo clinico do rosoxacin - Win-35213no tratamento da gonococcia aguda na mulher. / Clinical trial with rosoxacin Win-35213 - in the treatment of female acute gonorrhoeae

O autor estudou 26 casos de mulheres portadoras de gonococcia aguda, analisando a sintomatologia, diagnostico e tratamento com o rosoxacin (WIN-35213). Observou um indice de cura de 96,2% com uso de dose unica de 300 mg de rosoxacin por via oral. Os efeitos colaterais constatados, principalmente tontura e cefaleia, apresentaram regrassao espontanea