Analysis of bovine rotavirus strains circulating in diarrheic dairy calves in Uberaba, Minas Gerais, Brazil, during 2008-2009 / Análise de cepas de rotavírus bovino circulantes em bezerros leiteiros com diarreia em Uberaba, Minas Gerais, Brasil, durante 2008-2009

O objetivo deste estudo foi detectar rotavírus em fezes de bezerros com diarreia em Uberaba, MG, e caracterizar os genes VP7 e VP4 por meio da genotipagem e da análise filogenética. Setenta e quatro amostras foram coletadas entre novembro de 2008 e setembro de 2009. A detecção do vírus foi feita por teste de aglutinação e as amostras positivas foram submetidas à transcrição reversa, seguida de reação em cadeia da polimerase (RT-PCR), tipagem por PCR e sequenciamento. A taxa de detecção de rotavírus foi de 6,8% e todas as amostras apresentaram o genótipo G6P[5]. A análise filogenética mostrou que as amostras do genótipo G6 pertencem à linhagem IV e que, para ambos os genes (VP7 e VP4), as amostras deste estudo compõem um sub-cluster à parte daquele das cepas referências e das amostras campo mais similares. O alinhamento das sequências de aminoácidos deduzidas mostrou substituições em regiões antigênicas quando comparadas com as sequências das cepas bovinas UK e NCDV, presentes nas vacinas disponíveis no Brasil. Uma nova sublinhagem genética de G6P[5] foi evidenciada neste estudo. Substituições de aminoácidos nas regiões antigênicas dos rotavírus e a circulação de novas variantes podem representar desafios para as vacinas utilizadas atualmente. O presente estudo contribui para a compreensão da epidemiologia dos rotavírus bovinos no Brasil.(AU)

X-ray pelvimetry in common marmoset (Callithrix jacchus - LINNAEUS, 1758) / Pelvimetria radiográfica em sagui-de-tufo-branco (Callithrix jacchus - LINNAEUS, 1758)

The common marmoset is an important Neotropical primate for biomedical research. With the intention of providing pelvimetric values that allow the selection of animals with better characteristics for reproduction, we studied the X-ray of the pelvis, in ventro-dorsal projection of 12 adult animals, six males and six females, belonging to the National Center of Primates - CENP. The measurements were made to right diagonal diameters (RDD), left diagonal diameter (LDD), bi-iliac middle diameter (BIMD), bi-iliac upper diameter (BIUD), bi-iliac lower diameter (BILD), sacrum-pubic diameter (SPD), and the input area of the pelvis (IAP), which were compared in relation to sex and bodily correlated with the biometrics. The results of the means and standard deviations regarding pelvimetry (cm) were: for the RDD in males of 1.91±0.14 and in the females of 2.05±0.08; LDD in males of 1.85±0.16 and in females of 2.03± 0.08; BIMD in males of 1.46±0.12 and in the females of 1.5±0; BIUD showed in males 1.3±0.1 and in females of 1.3±0.06; BILD in males of 1.35±0.15 and in females of 1.25±0.08; SPD in males of 1.68±0.09 and in females of 1.93±0.13; IAP in males of 1.9±0.26 and in the females of 2.23±0.13. It was concluded that the pelvis of Callithrix jacchus could be classified as dolicopelvic type, there is sexual dimorphism in relation to the pelvis of adult animals and that the bodily measurements have low positive correlation with pelvic measurements.(AU)

O sagui-de-tufo-branco se destaca como um importante primata neotropical para as pesquisas biomédicas. Com o intuito de fornecer valores pelvimétricos que permitam a seleção de animais com melhores características para a reprodução, estudaram-se radiografias da pelve, em projeção ventrodorsal, de 12 animais, sendo seis machos e seis fêmeas, adultos, pertencentes ao Centro Nacional de Primatas - CENP. As mensurações realizadas foram referentes aos diâmetros diagonal direito (DDD), diagonal esquerdo (DDE), bi-ilíaco médio (DBIM), bi-ilíaco superior (DBIS), bi-ilíaco inferior (DBII), sacropúbico (DSP), além da área de entrada da pelve (AEP), os quais foram comparados em relação ao sexo e correlacionados com a biometria corpórea. Os resultados de médias e desvios-padrão com relação à pelvimetria (cm) foram: para o DDD nos machos de 1,91±0,14 e nas fêmeas de 2,05±0,08; DDE nos machos de 1,85±0,16 e nas fêmeas de 2,03±0,08; DBIM nos machos de 1,46±0,12 e nas fêmeas de 1,5±0; DBIS nos machos de 1,3±0,1 e nas fêmeas de 1,3±0,06; DBII nos machos de 1,35±0,15 e nas fêmeas de 1,25±0,08; DSP nos machos de 1,68±0,09 e nas fêmeas de 1,93±0,13; AEP nos machos de 1,9±0,26 e nas fêmeas de 2,23±0,13. Concluiu-se que a pelve de Callithrix jacchus pode ser classificada como do tipo dolicopélvica, existe dimorfismo sexual em relação às pelves de animais adultos e que as medidas corpóreas apresentam baixa correlação positiva com as medidas pélvicas.(AU)

Electroretinography and immunohistochemistry of retina in rabbits treated with sildenafil citrate / Eletrorretinografia e imuno-histoquímica da retina de coelhos tratados com citrato de sildenafil

Sildenafil citrate is a type-5 phosphodiesterase inhibitor (PDE-5), able to inhibit type-6 phosphodiesterase (PDE-6) as well, providing clinical benefits and paraeffects, some of them potentially related to the retina. The effects of the sildenafil on the retrobulbar and retinal circulation were studied in 27 adult male rabbits of the White New Zealand breed. The electric activity of the retina was evaluated before and at the end of the treatments, and immunohistochemistry studies were conducted. An amplitude increase of the b wave was found in the mixed response of cones and rods after 7 days of treatment with sildenafil citrate. However, in the other evaluations and periods, the values did not differ from the basal ones. Through immunohistochemistry, no significant decrease of the expression of PDE-5 and PDE-6 proteins was observed. Based on the results obtained, it is possible to admit that the sildenafil citrate did not change the expression of PDE-5 and PDE-6, neither the electroretinographic activity of the retina of male rabbits of the White New Zealand breed.

O citrato de sildenafil é um inibidor da fosfodiesterase do tipo 5 (PDE-5), capaz de inibir também a fosfodiesterase do tipo 6 (PDE-6), proporcionando benefícios clínicos e paraefeitos, alguns deles potencialmente relacionados à retina. Foram estudados efeitos do sildenafil sobre a circulação retrobulbar e a retiniana em 27 coelhos machos adultos, da raça Nova Zelândia Branco. Avaliou-se a atividade elétrica da retina antes e ao término dos tratamentos e realizaram-se estudos à imunoistoquímica. Encontrou-se aumento da amplitude da onda b na resposta mista de cones e de bastonetes, após 7 dias de tratamento com citrato de sildenafil. Entretanto, nas demais avaliações e períodos, os valores não divergiram dos basais. Pela imunoistoquímica, não se observou diminuição significativa da expressão das proteínas PDE-5 e PDE-6. Com base nos resultados obtidos, é possível admitir que o citrato de sildenafil não alterou a expressão de PDE-5 e PDE-6, tampouco, a atividade eletrorretinográfica da retina de coelhos machos da raça Nova Zelândia Branco.

Association of TNF -1031 C/C as a potential protection marker for leprosy development in Amazonas state patients, Brazil.

Polymorphisms present in the TNF promoter region has shown to influence the gene transcription. Leprosy displays different clinical manifestations according to the immune responses of the individual infected with Mycobacterium leprae. In this study, we evaluated the single nucleotide polymorphisms (SNPs) -238 G/A (rs361525), -308 G/A (rs1800629), -857 C/T (rs1799724), -863 A/C (rs1800630) and -1031 T/C (rs1799964) in the promoter region of the TNF to see whether these SNPs influence host-susceptibility to leprosy and the different clinical manifestation. Nucleotide sequencing was performed with DNA samples from 108 leprosy patients and 253 control subjects. An association between -1031 C/C genotype and protection from leprosy was observed when leprosy patients were compared to controls (OR 0.11; 95% CI=0.01-0.82; p=0.012). The -857 C/T genotype may be associated with susceptibility to leprosy (OR=1.81; 95% CI=1.09-3.00; p=0.028). Similar genotype and allele frequencies for the SNPs -308 G/A and -238 G/A were observed between leprosy patients and control subjects. Altogether, TNF polymorphisms in the promoter region may be predictive of leprosy outcome.

Malignant syphilis in an AIDS patient.

Malignant syphilis is an uncommon, but not unknown, ulcerative variation of secondary syphilis. The lesions typically begin as papules, which quickly evolve to pustules and then to ulcers with elevated edges and central necrosis. It is usually, but not mandatory, found in patients with some level of immunosuppression, such as HIV patients, when the TCD4(+) cell count is >200 cells/mm(3). Despite the anxiety the lesions cause, this form of the disease has a good prognosis. The general symptoms disappear right after the beginning of treatment, and lesions disappear over a variable period. This study reports the case of a 27-year-old man who has been HIV positive for 6 years, uses antiretroviral therapy incorrectly, has a TCD4(+) cell count of 340 cells/mm(3), a VDRL of 1:128 and itchy disseminated hyperchromic maculopapular lesions with rupioid crusts compatible with malignant syphilis.

Probiotic technological and functional characteristics of Lactobacillus strains isolated from chicken gut / Características tecnológicas e funcionais de linhagens de Lactobacillus isoladas do trato gastrointestinal de frangos

Lactobacillus spp. isolated from different portions of chickens' gastrointestinal tract were evaluated concerning their ability to survive in a water-in-oil (W/0) emulsion containing sesame and sunflower oil. After sixty days of emulsion storage under refrigeration, three of five strains tested survived in number equal to or higher than 10(6)cfu/g. Lactobacillus reuteri 2M14C, which presented the highest survival in W/O emulsion (10(7)cfu/g), was tested for its capacity to resist throughout the passage through gnotobiotic mice gastrointestinal tract and for the ability to stimulate murine peritoneal macrophages phagocytosis. This strain remained at a number above 10(9)cfu/g feces during ten days of monoassociation, and monoassociated mice showed phagocytic activity significantly greater than the germ-free controls (P<0.05). The results suggest that the formulation can be used to incorporate viable Lactobacillus spp. cells in animal feed. Moreover, the results suggest that L. reuteri 2M14C is a strong candidate to be incorporated in probiotic formulations for use in chicken.

Lactobacillus spp. isolados de diferentes porções do trato gastrintestinal de frangos foram testados quanto à capacidade de se manterem viáveis em uma emulsão água/óleo (A/O) contendo óleos de gergelim e de girassol. Após sessenta dias de estocagem sob refrigeração, três de cinco linhagens testadas sobreviveram em concentração igual ou superior a 10 6 UFC/g. Lactobacillus reuteri 2M14C, que apresentou maior capacidade de sobrevivência na emulsão desenvolvida (10 7 UFC/g), foi testado quanto à sua capacidade de sobreviver às condições do trato gastrintestinal in vivo em camundongos gnotobióticos. Após dez dias de monoassociação com L. reuteri 2M14C, foi testada também a capacidade de estimulação da atividade fagocítica de macrófagos peritoneais. A linhagem permaneceu em número superior a 10 9 UFC/g de conteúdo fecal durante os dez dias de monoassociação, e os camundongos monoassociados apresentaram atividade fagocítica maior (P<0,05) que a do grupo controle isento de germe. Os resultados sugerem que a formulação proposta é capaz de manter a viabilidade de células de lactobacilos para adição em ração animal, necessitando, no entanto, de um acompanhamento dessa viabilidade por tempo maior de estocagem. Além disso, os resultados demonstram que Lactobacillus reuteri 2M14C é um forte candidato a ser adicionado em formulações probióticas para uso em frangos.

Impairment of locomotor activity induced by the novel N-acylhydrazone derivatives LASSBio-785 and LASSBio-786 in mice

The N-acylhydrazone (NAH) analogues N-methyl 2-thienylidene 3,4-benzoylhydrazine (LASSBio-785) and N-benzyl 2-thienylidene 3,4-benzoylhydrazine (LASSBio-786) were prepared from 2-thienylidene 3,4-methylenedioxybenzoylhydrazine (LASSBio-294). The ability of LASSBio-785 and LASSBio-786 to decrease central nervous system activity was investigated in male Swiss mice. LASSBio-785 or LASSBio-786 (30 mg/kg, ip) reduced locomotor activity from 209 ± 26 (control) to 140 ± 18 (P < 0.05) or 146 ± 15 crossings/min (P < 0.05), respectively. LASSBio-785 (15 or 30 mg/kg, iv) also reduced locomotor activity from 200 ± 15 to 116 ± 29 (P < 0.05) or 60 ± 16 crossings/min (P < 0.01), respectively. Likewise, LASSBio-786 (15 or 30 mg/kg, iv) reduced locomotor activity from 200 ± 15 to 127 ± 10 (P < 0.01) or 96 ± 14 crossings/min (P < 0.01), respectively. Pretreatment with flumazenil (20 mg/kg, ip) prevented the locomotor impairment induced by NAH analogues (15 mg/kg, iv), providing evidence that the benzodiazepine (BDZ) receptor is involved. This finding was supported by the structural similarity of NAH analogues to midazolam. However, LASSBio-785 showed weak binding to the BDZ receptor. LASSBio-785 or LASSBio-786 (30 mg/kg, ip, n = 10) increased pentobarbital-induced sleeping time from 42 ± 5 (DMSO) to 66 ± 6 (P < 0.05) or 75 ± 4 min (P < 0.05), respectively. The dose required to achieve 50% hypnosis (HD50) following iv injection of LASSBio-785 or LASSBio-786 was 15.8 or 9.5 mg/kg, respectively. These data suggest that both NAH analogues might be useful for the development of new neuroactive drugs for the treatment of insomnia or for use in conjunction with general anesthesia.

Stem cell and tissue engineering therapies for ocular regeneration.

The eye is a relatively small but very complex organ. It is responsible for vision. Most of its cells are terminally differentiated, and several pathologies affecting those cells lead to vision loss and eventual blindness. Several years ago, a group of cells, located in the limbus, was identified as having the capacity of self-renewal and later on found to feed the renewal of the corneal epithelial layer. Since then, this niche of stem cells has been studied in order to provide clues that can be valuable for the regeneration of ocular structures. The worldwide shortage of donors, increased risk of transmissible diseases and immune rejection and the increased life expectancy, all contributed for the development of strategies to regenerate or repair ocular tissues. In this review we focus on two approaches for ocular regeneration: one based on stem cells and the other one based on tissue engineering strategies, and present examples where these two strategies overlap. We review the sources of cells and tissue engineering strategies for the regeneration of the cornea and of the retina, summarizing the most relevant and recent findings.

The role of iNOS and PHOX in periapical bone resorption.

Nitric oxide (NO) and reactive oxygen species (ROS) are key molecules in resistance to pathogens. Little is known about their role in pathogenesis of periapical lesions. To address this issue, we induced periapical lesions in mice lacking nitric oxide synthase (iNOS(-/-)) or phagocyte oxidase (PHOX(-/-)). iNOS(-/-) mice expressed higher levels of IL-1ß, TNF-α, RANK, RANKL, and MCP-1 than C57BL/6 and PHOX(-/-). Apical thickening of the periodontal ligament was also greater in iNOS(-/-) compared with other groups. Interestingly, ROS production did not interfere in periapical lesion progression, but seemed to be essential for the appearance of multinucleated TRAP-positive cells. Thus, periapical lesion progression in iNOS(-/-) was associated with an imbalance of pro-inflammatory cytokines (IL-1ß and TNF-α), bone-resorptive modulators (RANK and RANKL), and MCP-1. We conclude that NO, but not ROS, controls progression of bone resorption in a murine experimental model of apical periodontitis.

The role of tumor necrosis factor receptor type 1 in orthodontic tooth movement.

Orthodontic tooth movement is dependent on osteoclast activity. Tumor necrosis factor (TNF)-alpha plays an important role, directly or via chemokine release, in osteoclast recruitment and activation. This study aimed to investigate whether the TNF receptor type 1 (p55) influences these events and, consequently, orthodontic tooth movement. An orthodontic appliance was placed in wild-type mice (WT) and p55-deficient mice (p55(-/-)). Levels of TNF-alpha and 2 chemokines (MCP-1/CCL2, RANTES/CCL5) were evaluated in periodontal tissues. A significant increase in CCL2 and TNF-alpha was observed in both groups after 12 hrs of mechanical loading. However, CCL5 levels remained unchanged in p55(-/-) mice at this time-point. The number of TRAP-positive osteoclasts in p55(-/-) mice was significantly lower than that in WT mice. Also, there was a significantly smaller rate of tooth movement in p55(-/-) mice. Analysis of our data suggests that the TNFR-1 plays a significant role in orthodontic tooth movement that might be associated with changes in CCL5 levels.

Efeitos agudos dos extratos hidroalcoólicos do alho (Allium sativum L.) e do capim-limão (Cymbopogon citratus (DC) Stapf) sobre a pressão arterial média de ratos anestesiados / Acute effects of Allium sativum L. and Cympobongon citratus (DC) Stapf hydroalcoholic extracts on arterial blood pressure of anesthetized rats

Este trabalho objetivou verificar os efeitos da aplicação aguda endovenosa dos extratos hidroalcoólicos de Allium sativum e de Cymbopogon citratus sobre a pressão arterial de ratos. Foram usados Rattus novergicus albinus, n=7, anestesiados, traqueostomizados e canulados através da veia jugular e da artéria carótida. Foram injetadas doses de 1 mg dos extratos separadamente e em associação (1mg + 1mg), em volumes de 0,2mL. A pressão arterial média (PAM) foi registrada com um sistema Biopac, modelo MP100. O Allium sativum diminuiu a PAM de 124±2 mmHg, no controle, para 108±2 mmHg aos 15s. Da mesma forma, o Cymbopogon citratus diminuiu a PAM de 122±2 mmHg, no controle, para 106± 2 mmHg aos 15s. A associação de ambos também diminuiu a PAM de 126±3 mmHg, no controle, para 113±3 mmHg aos 15s. Os efeitos das duas plantas foram iguais e não foram incrementados quando associadas.

This work aimed to verify the effects of acute intravenous applications of Allium sativum and Cymbopogon citratus hydroalcoholic extracts on arterial blood pressure of anesthetized rats. Adult male rats (Rattus novergicus albinus), n=7, were used. The rats were anesthetized, tracheotomy and cannulation of both jugular and carotid were carried out. The injected doses were 1 mg separately as well as the association of both extracts, in volumes of 0,2 mL. The mean arterial blood pressure (MAP) was recorded with a Biopac System, model MP100. The Allium sativum decreased the MAP only from 124±2 mmHg (control) to 108±2 mmHg at 15s. The Cymbopogon citratus also decreased the MAP from 122±2 mmHg (control) to 106±2 mmHg after 15s. The 1mg of Allium sativum + 1mg of Cymbopogon citratus also decreased the MAP from 126±3 mmHg (control) to 113±3 mmHg after 15s. The effects of the two plants were the same and were not increased when in association.

Atividade antimicrobiana de Struthanthus vulgaris (erva-de-passarinho) / Antimicrobial activity of Struthantus vulgaris (erva-de-passarinho)

As plantas do gênero Struthanthus são conhecidas como ervas-de-passarinho e parasitam pomares no Brasil, principalmente os de laranjeiras e goiabeiras. Na medicina popular são usadas nas afecções das vias respiratórias. O extrato hidroetanólico a 70 por cento de folhas frescas de Struthanthus vulgaris apresentou atividade antimicrobiana contra amostras bacterianas Gram positiva e Gram negativa. Este extrato não apresentou, nas condições testadas, atividade contra fungos. As amostras bacterianas mais sensíveis ao extrato foram Bacillus cereus (ATCC 11778), Micrococcus luteus (ATCC 9341), Staphylococcus aureus (ATCC 6538), S. epidermidis (ATCC 12228) e P. aeruginosa (ATCC 27853), usando o método de difusão em agar. As frações obtidas, pela partição líquido-líquido do extrato hidroetanólico a 70 por cento, com solventes de polaridades crescentes (clorofórmio, acetato de etila, n-butanol e água), apresentaram diferentes atividades inibitórias. A fração que apresentou a maior atividade contra bactéria Gram positiva (B. cereus) e Gram negativa (P. aeruginosa) foi aquela obtida com n-butanol. Nessa fração foram detectados flavonóides, taninos condensados (proantocianidinas) e saponinas.

Struthantus vulgaris (mistletoe) is one of the most common hemiparasite species in Brazil. It occurs as a parasite of orchards, mainly in orange and guava trees. Some authors mention Struthantus use in traditional medicine for respiratory diseases treatment. Fresh leaves concentrated hydroalcoholic extract showed antimicrobial activity against Gram positive and Gram negative bacterial samples. In tested conditions, these extracts did not show activity against fungi. The more susceptible bacterial samples to fresh leaves hydroalcoholic extract were Bacillus cereus (ATCC 11778), Micrococcus luteus (ATCC 9341), Staphylococcus aureus (ATCC 6538), S. epidermidis (ATCC 12228) and Pseudomonas aeruginosa (ATCC 27853). The method used for assessment of antimicrobial activity was agar diffusion. Fractions obtained from fresh leaves concentrated alcoholic extracts with increasing polarity solvents (chloroform, ethyl acetate, n-butanol and water) showed different inhibitory activities. n-Butanol fraction showed greater activity against Gram positive bacteria (B. cereus) and Gram negative bacteria (P. aeruginosa). In this fraction, flavonoids, condensed tannins (proanthocyanidins) and saponins, were found.

[Obesity and malnutrition in children: profile of a low-income community] / Desnutrição e obesidade em crianças: delineamento do perfil de uma comunidade de baixa renda.

OBJECTIVE: To establish the nutritional diagnosis of children living in a low-income community through anthropometric evaluation, detecting insufficient nutrition (low weight-for-age, wasting and stunting) and excessive nutrition (overweight and obesity). METHODS: A community-based survey identified children below 11 years living in a low-income community. Their weight and height (or lengths) were measured. The National Center for Health Statistics growth curve was used as reference. The cut-off points used were: low-weight-for-age z score of < -2 (weight-for-age), wasting (weight-for-height) and stunting (height-for-age); respective overweight and obesity z scores of +1 to +2 and of > +2, according to weight-for-height. RESULTS: Prevalence rates were 3.8% for low weight for age, and 24% for low-weight-for-age risk factors; 1.2% for wasting, and 21.6% for wasting risk factors; 5% for stunting, and 14.3% for stunting risk factors; 10.1% for overweight, and 4.6% for obesity. CONCLUSIONS: Main nutritional issues were present in children from that community, with simultaneous occurrence of chronic malnutrition and obesity.

[Study of lung involvement in patients with cystic fibrosis] / Estudo do quadro pulmonar de pacientes com fibrose cística.

OBJECTIVE: To characterize the involvement of the respiratory apparatus of patients with cystic fibrosis in order to obtain a comprehensive view of their pulmonary picture.METHODS: Data were obtained retrospectively from the medical records of 16 patients with cystic fibrosis; arterial gas and spirometry data were obtained prospectively for the same patients, who were not in an acute pulmonary situation. The patients were subjects of both sexes aged 6 years or older who were followed up at the Pediatrics Outpatient Clinic of the University Hospital, Faculty of Medicine of Ribeirão Preto, USP.RESULTS: Median patient age was 114 months (9 years and 6 months) ranging between 72 - 360 months, and 68.75% were males. Productive cough was the most frequent symptom observed in 75% of the population studied. All patients had positive sputum culture obtained at least one year before, with Pseudomonas aeruginosa being detected in 81.25% of the cases. Arterial gases revealed some abnormalities in 81.25% of the patients and spirometry revealed abnormalities in 56.25%.CONCLUSION: All patients presented at least one type of pulmonary alteration. Measurement of arterial gases detected a larger number of patients with altered pulmonary function than did spirometry, but the two examinations complemented each other for a good evaluation of pulmonary function.

Seroprevalência da infecçäo pelo HIV-1 em gestantes atendidas no IMIP, Recife - Brasil / Seroprevalence of HIV infection among pregnant women attending in the IMIP, Recife-Brasil

The number of HIV infected heterosexual has been steadily increasing in Brazil. This fact was followed by a decline on the ratio of male/female HIV infection every year. We have examined the seroprevalence of HIV infection among pregnant women attending an antenatal clinic in the Instituto Materno Infantil de Pernambuco (IMIP) at Recife, North-eastern Brazil. This study was performae as an anonymous sentinel surveillance. The collected blood samples were tested for HIV infection by two different types of ELISA, and if positive, further analysis by IIA was submitted. The subjects were arranged in four age groups with the respective proportion: group l (<15 years)-0.9 per cent, group II (15 to 20 years)-31.6 per cent, group III (21 to 34 years)-61.7 per cent and group IV (>35 years)- 5,8 per cent. It was found an HIV-1 seroprevalence of 0.1 per cent (95 per cent confidence interval, -0.l to + 0,3) This percentage represents one single HIV positive woman included in age group III. his low HIV infection seroprevalence is suitable with a low risk population (pregnant women) in a low prevalence area. However, we became concerned with the great propottion of pregnant woman under 21 years (32,5 per cent) and with the implications of how to reinforce effectively HIV preventions of how to reinforce effectively HIV prevention strategies to this population

Cystatin S in secretory granules fractions isolated from submandibular gland of infected rats by Trypanosoma cruzi.

We have measured the relative concentrations of cysteine proteinase in the granular fractions of submandibular glands of control and Trypanosoma cruzi infected rats by using a sensitive solid phase ELISA. Gland samples were homogenized in 0.34 M sucrose and 0.5 mM EDTA in 10 mM HEPES buffer at a pH of 7.4. The extract was centrifuged and filtered through Millipore filters to prepare a purer granular fraction. Immunochemical studies using antibody against cystatin S and electrophoretic analysis showed higher cystatin S levels in infected rats than in control. The role of this inhibitor during acute phase of Chagas disease is discussed.

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).

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).