RESUMO O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.

ABSTRACT The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.

Abstract: Medicinal plants have long been used as an alternative to traditional drugs for the treatment of inflammatory conditions due to the classical side effects and restricted access of various commercially available drugs, such as steroids (GCs) and nonsteroidal anti-inflammatory drugs (NSAIDs). Sambucus australis is a Brazilian herb that is commonly used to treat inflammatory diseases; however, few studies have examined the use of this species in the treatment of inflammatory conditions. The present study aims to evaluate the potential anti-inflammatory activity of S. australis in vitro. We established spleen cell cultures stimulated with pokeweed mitogen (PWM) to evaluate the production of proinflammatory cytokines, such as IL-4, IL-5, IFN-y, and IL-10 (by ELISA), and the expression of the transcription factor NF-kB (by RT-PCR). In addition, we evaluated the levels of nitric oxide in macrophage cultures and the membrane-stabilizing activity of S. australis methanolic extract (EMSA). Treatment with EMSA at concentrations of 100, 50, 25 and 12.5 µg/ml significantly decreased IL-4 (p<0.001) and IL-5 (p<0.001) levels. Treatment with 100 µg/ml EMSA reduced IFN-у (p<0.001) levels. Moreover, at 100 mg/ml, EMSA also increased IL-10 production and reduced NF-kB expression (p<0.01). In macrophage cultures stimulated with LPS, EMSA decreased nitric oxide levels (p<0.001) at all concentrations tested (100, 50, 25 and 12.5 µg/ml). Additionally, EMSA had a protective effect in the erythrocyte membrane stabilization assay. Taken together, these results suggest that S. australis has anti-inflammatory potential in vitro, characterized by the reduction of both inflammatory cytokines and the expression of NF-kB along with the up-regulation of IL-10.

ABSTRACT This study evaluated the chemical composition and antioxidant activity of fatty acids from the marine red algae Pterocladiella capillacea (S. G. Gmelin) Santelices & Hommersand 1997 and Osmundaria obtusiloba (C. Agardh) R. E. Norris 1991. The gas chromatography mass spectrometry (GC-MS) identified nine fatty acids in the two species. The major fatty acids of P. capillacea and O. obtusiloba were palmitic acid, oleic acid, arachidonic acid and eicosapentaenoic acid. The DPPH radical scavenging capacity of fatty acids was moderate ranging from 25.90% to 29.97%. Fatty acids from P. capillacea (31.18%) had a moderate ferrous ions chelating activity (FIC), while in O. obtusiloba (17.17%), was weak. The ferric reducing antioxidant power (FRAP) of fatty acids from P. capillacea and O. obtusiloba was low. As for β-carotene bleaching (BCB), P. capillacea and O. obtusiloba showed a good activity. This is the first report of the antioxidant activities of fatty acids from the marine red algae P. capillacea and O. obtusiloba.

Abstract Ximenia americana L., Olacaceae, barks are utilized in folk medicine as analgesic and anti-inflammatory. The objective was to evaluate the toxicity and antinociceptive effect of polysaccharides rich fractions from X. americana barks. The fractions were obtained by extraction with NaOH, followed by precipitation with ethanol and fractionation by ion exchange chromatography. They were administered i.v. or p.o. before nociception tests (writhing, formalin, carragenan-induced hypernociception, hot plate), or during 14 days for toxicity assay. The total polysaccharides fraction (TPL-Xa: 8.1% yield) presented 43% carbohydrate (21% uronic acid) and resulted in two main fractions after chromatography (FI: 12%, FII: 22% yield). FII showed better homogeneity/purity, content of 44% carbohydrate, including 39% uronic acid, arabinose and galactose as major monosaccharides, and infrared spectra with peaks in carbohydrate range for COO- groups of uronic acid. TPL-Xa (10 mg/kg) and FII (0.1 and 1 mg/kg) presented inhibitory effect in behavior tests that evaluate nociception induced by chemical and mechanical, but not thermal stimuli. TPL-Xa did not alter parameters of systemic toxicity. In conclusion, polysaccharides rich fractions of X. americana barks inhibit peripheral inflammatory nociception, being well tolerated by animals.

RESUMO No Brasil, estudos apontam a circulação de Neorickettsia risticii em equinos, contudo não estão claros quais os possíveis vetores intermediários dessa bactéria no país. O objetivo do presente estudo foi analisar a presença de N. risticii, utilizando-se técnicas moleculares, em caramujos e estágios larvais de trematódeos em propriedades rurais de uma região com histórico de equinos sororreativos para essa bactéria, no Rio de Janeiro, Brasil. Uma amostragem por conveniência foi utilizada na região de estudo. Os caramujos coletados foram expostos à luz incandescente (60W) durante duas-quatro horas para a investigação de trematódeos nas formas larvais. A extração de ácido desoxirribonucleico (DNA) foi realizada em tecidos de caramujos e trematódeos. A técnica de PCR em tempo real (qPCR) foi utilizada para investigar a presença de um fragmento do gene 16S rRNA de N. risticii. Foram coletados 410 espécimes de caramujos de 11 propriedades com criações de equinos, sendo identificadas as seguintes espécies: Melanoides tuberculata, Pomacea sp., Biomphalaria tenagophila, Physa acuta, Drepanotrema anatinum e Biomphalaria straminea. Apenas 3,17% (n=13/410) dos caramujos identificados estavam infectados por trematódeos. As cercárias obtidas desses caramujos foram classificadas em Megalourous cercariae, Pleurolophocercus cercariae e Furcocercous cercariae. Não foi observada a amplificação do DNA-alvo de N. risticii, por meio da qPCR, em nenhuma das amostras de caramujos e trematódeos testadas. Com base nesses dados, a transmissão de N. risticii por trematódeos que utilizam as espécies de caramujos nessa região parece não ocorrer ou ocorre a taxas muito reduzidas. Portanto, novos estudos são necessários para elucidar quais espécies de hospedeiros invertebrados se infectam por essa bactéria e potencialmente participam da cadeia de transmissão da neorickettsiose equina no estado do Rio de Janeiro, Brasil.

ABSTRACT In Brazil, some studies have indicated that Neorickettsia risticii circulates in horses, but it is unclear which are the possible intermediate vectors of this bacterium in the country. The aim of this study was to use molecular techniques in order to analyze the presence of N. risticii in snails and larval stages of trematodes in farms in a region with a history of seroreactive horses towards this bacterium, in Rio de Janeiro, Brazil. Convenience sampling was used in the studied region. The collected snails were exposed to incandescent light (60W) for 2-4 hours in order to investigate trematodes in larval forms. Deoxyribonucleic acid (DNA) was extracted from snail tissue and trematode. Real-time PCR (qPCR) technique was used to investigate the presence of a 16S rRNA gene fragment of N. risticii. Snail specimens (n=410) were collected from 11 horse-breeding farms, and the following species were identified: Melanoides tuberculata, Pomacea sp., Biomphalaria tenagophila, Physa acuta, Drepanotrema anatinum and Biomphalaria straminea. Only 3.17% (n=13/410) of the collected snails were infected by trematodes. The cercariae obtained from these snails were classified as Megalourous cercariae, Pleurolophocercus cercariae and Furcocercous cercariae. There was no amplification of the target DNA of N. risticii in the snail and trematode samples tested by qPCR. Based on these data, the transmission of N. risticii by trematodes using these snail species in this region does not appear to occur or occurs at very low rates. Thus, further studies are needed in order to clarify which species of invertebrate hosts are infected by this bacterium and potentially participate in the transmission chain of equine neorickettsiosis in the state of Rio de Janeiro, Brazil.

Os invertebrados marinhos são capazes de sintetizar compostos bioativos que podem ser benéficos à saúde humana. O objetivo desse estudo foi avaliar as atividades antioxidante, hemolítica, antimicrobiana e citotóxica do extrato bruto (EtOH 70%) e frações em diclorometano (DMC), em acetato de etila (AcOEt) e aquosa (Aq) do zoantídeo marinho Palythoa caribaeorum. Os conteúdos fenólicos totais do extrato bruto e das frações DCM, AcOEt e Aq foram 12,33; 18,17; 10,53 e 3,18 mg AGE por grama, respectivamente. O sequestro do radical DPPH mostrou uma pequena variação. O IC50 do extrato bruto e das frações DCM, AcOEt e Aq foram 11,13; 11,25; 11,74 e 11,28 g mL-1, respectivamente. Dentre as amostras, a habilidade de quelação foi maior no extrato bruto (IC50 302,90 µg mL-1), seguida pelas frações AcOEt, Aq e DCM com 457,77; 547,91 e 641,82 µg mL-1, respectivamente. O poder de redução do ferro apresentou uma densidade óptica em torno de 0,5. As amostras testadas exibiram baixa atividade hemolítica, inferior a 10%, até a concentração de 50 µg mL-1. Não foi observada atividade antimicrobiana para as cepas bacterianas testadas. Para a atividade citotóxica, LC50 de DCM, extrato bruto, AcOEt e Aq foram, respectivamente, 52,10; 83,06; 86,34 e 117,45 µg mL-1, mostrando elevada toxicidade.

Marine invertebrates are capable of synthesizing bioactive compounds, which may be beneficial to human health. The aim of this study was to evaluate the antioxidant, hemolytic, antimicrobial and cytotoxic activities of crude extract (70% EtOH), and dichloromethane (DCM), ethyl acetate (EtOAc), and aqueous (Aq) fractions of the marine zoanthid Palythoa caribaeorum. The phenolic compound contents of the crude extract, DCM, EtOAc and Aq fractions were 12.33, 18.17, 10.53, and 3.18 mg GAE per gram, respectively. DPPH radical scavenging activity showed slight variation. IC50 of crude extract, DCM, EtOAc and Aq fractions were 11.13, 11.25, 11.74, and 11.28 µg mL-1, respectively. Among the sample, ferrous ion chelating was the highest in crude extract (IC50 302.90 µg mL-1), followed by EtOAc, Aq, and DCM fractions with 457.77, 547.91, and 641.82 µg mL-1, respectively. Ferric-reducing antioxidant power showed optical density at about 0.5. The samples tested exhibited low hemolytic activity under 10% up to a concentration of 50 μg mL-1. No antimicrobial activity was observed against any of the tested bacterial strains. For the cytotoxic activity, LC50 of DCM, crude extract, EtOAc, and Aq were 52.10, 83.06, 86.34, and 117.45 μg mL-1, showing high toxicity.

Antioxidantes naturais encontrados em macroalgas marinhas são substâncias bioativas conhecidas por desempenhar um papel importante na prevenção de doenças associadas com o envelhecimento, protegendo as células de danos oxidativos. O objetivo deste estudo foi avaliar as atividades antioxidante e citotóxica de extratos etanólicos de duas espécies de algas vermelhas, Amansia multifida eMeristiella echinocarpa. A atividade antioxidantein vitro foi determinada pela capacidade de sequestro do radical DPPH, o poder de redução do ferro (FRAP), o poder de quelação do íon ferroso (FIC), o teste de oxidação acoplada β-caroteno/ácido linoleico (BCB), bem como a quantificação do conteúdo fenólico total. A citotoxicidade foi avaliada pelo teste de letalidade contra Artemia sp. Os valores das substâncias fenólicas observados no presente estudo indicaram que as espéciesA. multifida e M. echinocarpa são ricas em substâncias fenólicas, atingindo valores de 45,40 e de 28,86 mg de ácido gálico equivalente (AGE) g−1de extrato etanólico, respectivamente. A capacidade de seqüestro do radical DPPH e o poder de quelação do íon ferroso foram 60% e 17%, respectivamente. Ambos os extratos algáceos inibiram a oxidação do β-caroteno em torno de 40%. Nenhum dos extratos algáceos foi potencialmente citotóxico. Os resultados mostraram que os extratos destas espécies de rodófitas apresentam potencial antioxidante e baixa toxicidade. Elas são fontes de compostos antioxidantes naturais.

Natural antioxidants found in marine macroalgae are bioactive compounds known to play an important role in the prevention of diseases associated with aging cells protecting them against the oxidative damage. The purpose of this study was to evaluate the antioxidant and cytotoxic activity of ethanolic extracts of two species of red seaweeds, Amansia multifida and Meristiella echinocarpa. In vitro antioxidant activity was determined by DPPH radical scavenging assay, ferric-reducing antioxidant power (FRAP) assay, ferrous ion chelating (FIC) assay, β-carotene bleaching (BCB) assay and total phenolic content (TPC) quantification. Cytotoxicity was evaluated with the brine shrimp Artemia sp. lethality test. The TPC values observed in the present study indicated that both species A. multifida and M. echinocarpa are rich in phenolic compounds, reaching values of 45.40 and 28.46 mg gallic acid equivalent (GAE) g−1 of ethanolic extract, respectively. DPPH radical scavenging and ferrous ion chelating showed values of 60% and 17%, respectively. Both seaweed extracts inhibited β-carotene oxidation by approximately 40%. None of the algal extracts were potentially cytotoxic. The results have showed that extracts of both species of marine red algae exhibit antioxidant potential and low toxicity. They are sources of natural antioxidant compounds.

OBJECTIVE: This study was conducted to investigate factors associated with thrombocytopenia in a large cohort of patients with leptospirosis in an endemic area. METHODS: This retrospective study included 374 consecutive patients with leptospirosis who were admitted to tertiary hospitals in Fortaleza, Brazil. All patients had a diagnosis of severe leptospirosis (Weil's disease). Acute kidney injury was defined according to the RIFLE criteria. Thrombocytopenia was defined as a platelet count <100,000/mm3. RESULTS: A total of 374 patients were included, with a mean age of 36.1±15.5 years, and 83.4% were male. Thrombocytopenia was present at the time of hospital admission in 200 cases (53.5%), and it developed during the hospital stay in 150 cases (40.3%). The patients with thrombocytopenia had higher frequencies of dehydration (53% vs. 35.3%, p = 0.001), epistaxis (5.7% vs. 0.8%, p = 0.033), hematemesis (13% vs. 4.6%, p = 0.006), myalgia (91.5% vs. 84.5%, p = 0.038), hematuria (54.8% vs. 37.6%, p = 0.011), metabolic acidosis (18% vs. 9.2%, p = 0.016) and hypoalbuminemia (17.8% vs. 7.5%, p = 0.005). The independent risk factors associated with thrombocytopenia during the hospital stay were lengthy disease (OR: 1.2, p = 0.001) and acute kidney injury (OR: 6.6, p = 0.004). Mortality was not associated with thrombocytopenia at admission (12.5% vs. 12.6%, p = 1.000) or during the hospital stay (12.6% vs. 11.3%, p = 0.748). CONCLUSIONS: Thrombocytopenia is a frequent complication in leptospirosis, and this condition was present in more than half of patients at the time of hospital admission. Lengthy disease and acute kidney injury are risk factors for thrombocytopenia. There was no significant association between thrombocytopenia and mortality.

OBJETIVOS: Avaliar o programa de rastreamento neonatal da Secretaria de Estado da Saúde de Santa Catarina, em relação ao hipotireoidismo congênito (HC), e estimar sua prevalência nas crianças rastreadas. MÉTODO: Foram rastreadas 390.759 crianças no período de 01/94 a 12/98, sendo avaliada: a cobertura do programa, as idades na coleta da 1ª amostra de sangue para dosagem de TSH, os tempos para envio da amostra ao laboratório central (LACEN), resultado da dosagem do TSH e localização das crianças com exames alterados e as idades na 1ª consulta e no início do tratamento e a dose de L-T4 prescrita. RESULTADOS: A cobertura do programa foi de 81%, tendo sido detectadas 123 crianças com HC, com prevalência de 1:3.177. A idade média das crianças na coleta da 1ª amostra foi de 17,6 dias. As médias de tempo foram: 7,4 dias para a chegada da amostra ao LACEN, 2,4 dias para o resultado da dosagem do TSH e 7,6 dias para a localização da criança e a 1ª consulta. Todas as crianças detectadas foram atendidas no Hospital Infantil Joana de Gusmão e tinham, em média, idade de 40,2 dias na 1ª consulta e no início do tratamento. A dose média de L-T4 prescrita foi 12,5µg/kg/dia. CONCLUSÃO: O tempo para a dosagem do TSH e a dose de L-T4 prescrita são adequados. As demais variáveis estão fora do tempo preconizado, acarretando atraso no início do tratamento. A prevalência de HC é de 1 caso para cada 3.177 crianças rastreadas.

OBJECTIVES: To evaluate the neonatal screening program for congenital hypothyrodism (CH) of the Health Secretariat of the State of Santa Catarina and determine the prevalence of CH among screened children. METHODS: 390,759 newborns were screened for CH between 01/94 and 12/98 to evaluate: the program coverage, children's age at the time of the 1st blood sample for TSH measurement, time to transport blood samples to the core laboratory (LACEN), time to obtain TSH results, time to locate children with abnormal exams, maternal age at the time of the 1st clinical appointment and beginning of treatment, and dose of L-T4 prescribed. RESULTS: Program coverage was 81% and 123 children were diagnosed with CH and the estimated prevalence was 1:3,177. The mean age at the time of the 1st blood sample was 17.6 days. The mean intervals between blood sampling and the various outcomes were: 7.4 days to transport samples to LACEN, 2.4 days to obtain laboratory results, 7.6 days to locate positive-screened children and bring them to their 1st clinical appointment. All positive-screened children were evaluated at Joana de Gusmão Sick Children Hospital. The mean age at the time of the 1st clinical appointment and beginning of treatment was 40.2 days. The mean dose of L-T4 prescribed was 12.5µg/kg/day. CONCLUSION: Time between TSH dosage and prescription of L-T4 is appropriate. All other variables are outside the recommended time frame, resulting in delays to initiate therapy. The prevalence of CH among screened children is 1:3,177.

Para testar a hipótese de que a população de baixo nível socioeconômico apresenta diferentes níveis de pobreza que repercutem em seu estado de saúde, foi realizado estudo transversal envolvendo 477 famílias de uma população urbana pobre residente na Vila Grande Cruzeiro, em Porto Alegre, RS, Brasil. O nível de pobreza dessas famílias foi medido por meio de um instrumento especialmente elaborado para populações urbanas pobres. Crianças oriundas de famílias vivendo em extrema pobreza (quartil inferior) apresentaram maior taxa de mortalidade infantil, menor peso de nascimento, maior número de internações hospitalares e maiores índices de desnutrição, além de pertencerem a famílias mais numerosas. A população de baixo nível socioeconômico mostrou-se heterogênea em relação a diferentes indicadores de saúde. Concluiu-se que a identificação de subgrupos mais vulneráveis numa população permite concentrar as ações de saúde entre os mais necessitados.

To test the hypothesis that the low socioeconomic population living is shanty towns in Porto Alegre presents different levels of poverty which are reflected on its health status, a cross-sectional study was designed involving 477 families living in Vila Grande Cruzeiro, Porto Alegre, Brazil. The poverty level of the families was measured by using an instrument specifically designed for poor urban populations. Children from families living in extreme poverty (poorest quartile) were found to have higher infant mortality rate, lower birth weights, more hospitalizations, and higher malnutrition rates, in addition to belonging to more numerous families. Thus, the shanty town population of Porto Alegre is not homogeneous, and priority should be given to the more vulnerable subgroups.

Para testar a hipótese de que a população de baixo nível socioeconômico apresenta diferentes níveis de pobreza que repercutem em seu estado de saúde, foi realizado estudo transversal envolvendo 477 famílias de uma população urbana pobre residente na Vila Grande Cruzeiro, em Porto Alegre, RS, Brasil. O nível de pobreza dessas famílias foi medido por meio de um instrumento especialmente elaborado para populações urbanas pobres. Crianças oriundas de famílias vivendo em extrema pobreza (quartil inferior) apresentaram maior taxa de mortalidade infantil, menor peso de nascimento, maior número de internações hospitalares e maiores índices de desnutrição, além de pertencerem a famílias mais numerosas. A população de baixo nível socioeconômico mostrou-se heterogênea em relação a diferentes indicadores de saúde. Concluiu-se que a identificação de subgrupos mais vulneráveis numa população permite concentrar as ações de saúde entre os mais necessitados.

To test the hypothesis that the low socioeconomic population living is shanty towns in Porto Alegre presents different levels of poverty which are reflected on its health status, a cross-sectional study was designed involving 477 families living in Vila Grande Cruzeiro, Porto Alegre, Brazil. The poverty level of the families was measured by using an instrument specifically designed for poor urban populations. Children from families living in extreme poverty (poorest quartile) were found to have higher infant mortality rate, lower birth weights, more hospitalizations, and higher malnutrition rates, in addition to belonging to more numerous families. Thus, the shanty town population of Porto Alegre is not homogeneous, and priority should be given to the more vulnerable subgroups.