SYNTHESIS AND BIOLOGICAL EVALUATION OF SPIRO[INDOLINE-3,4217;-PYRANO[2,3-C:6,5-C217;]DIPYRAZOL]- 2-ONES IN THE PRESENCE OF SBA-Pr-SO3H AS A NANOCATALYST

Sulfonic acid functionalized SBA-15 nanoporous material (SBA-Pr-SO3H) with a large pore size of 6 nm, a high surface area, high selectivity, and excellent chemical and thermal stability was applied as an efficient heterogeneous nanoporous acid catalyst in the reaction of isatin with pyrazolones under mild reaction conditions. A novel class of symmetrical spiro[indoline-3,4'-pyrano[2,3-c:6,5-c']dipyrazol]-2-one derivatives was successfully obtained in high yields. Comparison of these results with those reported in the literature shows that the current method is efficient, and results in better reaction times and yields of the desired products. Other advantages of this new method are its operational simplicity, easy work-up procedure, and the use of SBA-Pr-SO3H as a reusable and environmentally benign nanoreactor, such that the reaction proceeds easily in its nanopores. We also tested the antimicrobial activity of the prepared compounds using the disc diffusion method, and some of the synthesized compounds exhibited the best results against B. subtilis and S. aureus.

Evaluation of GBV-C / HVG viremia in HIV-infected women

The present study aimed at standardizing a real-time quantitative polymerase chain reaction assay to evaluate the presence of GBV-C/HGV RNA. A "TaqMan" assay using primers and probe derived from the 5¢ NCR region was developed and validated. Two hundred and fifty-three plasma samples from HIV-infected women were tested for GBV-C viremia and antibody against the envelope protein 2. GBV-C RNA was detected in 22.5% of the patients whereas the antibody was identified in 25.3% of the cohort. Detection of viral RNA and of antibodies was mutually exclusive. Viral loads showed a mean of 1,777 arbitrary units / mL, being 1.1 and 13,625 arbitrary units / mL respectively the lowest and highest values measured. We conclude that the real-time quantitative polymerase chain reaction method developed is appropriate for the investigation of GBV-C RNA since it was shown to be highly specific and sensitive, as well as requiring few steps, preventing contamination and providing additional information as to the relative viremia of carriers, a parameter that must be included in studies evaluating the co-factors influencing the clinical outcome of HIV/AIDS.

Experimental toxoplasmosis in Balb/c mice. Prevention of vertical disease transmission by treatment and reproductive failure in chronic infection

In a study of congenital transmission during acute infection of Toxoplasma gondii, 23 pregnant Balb/c mice were inoculated orally with two cysts each of the P strain. Eight mice were inoculated 6-11 days after becoming pregnant (Group 1). Eight mice inoculated on the 10th-15th day of pregnancy (Group 2) were treated with 100 mg/kg/day of minocycline 48 h after inoculation. Seven mice inoculated on the 10th-15th day of pregnancy were not treated and served as a control (Group 3). Congenital transmission was evaluated through direct examination of the brains of the pups or by bioassay and serologic tests. Congenital transmission was observed in 20 (60.6%) of the 33 pups of Group 1, in one (3.6%) of the 28 pups of Group 2, and in 13 (54.2%) of the 24 pups of Group 3. Forty-nine Balb/c mice were examined in the study of congenital transmission of T. gondii during chronic infection. The females showed reproductive problems during this phase of infection. It was observed accentuated hypertrophy of the endometrium and myometrium. Only two of the females gave birth. Our results demonstrate that Balb/c mice with acute toxoplasmosis can be used as a model for studies of congenital T. gondii infection. Our observations indicate the potential of this model for testing new chemotherapeutic agents against congenital toxoplasmosis.

Expression of c-erbB-2, p53 and c-myc proteins in male breast carcinoma: Comparison with traditional prognostic factors and survival

There are few data evaluating biological markers for men with breast cancer. The purpose of the present study was to analyze the expression of the oncogenes c-erbB-2 and c-myc and of the suppressor gene p53 by immunohistochemical techniques in archival paraffin-embedded tissue blocks of 48 male breast cancer patients, treated at the A.C. Camargo Cancer Hospital, São Paulo, SP, Brazil. The results were compared with clinicopathological prognostic features. Immunopositivity of c-erbB-2, p53 and c-myc was detected in 62.5, 16.7 and 20.8% of the cases analyzed, respectively. Estrogen and progesterone receptors were positive in 75 and 69% of the cases, respectively. Increasing staging was statistically associated with c-erbB-2 (P = 0.04) and weakly related to p53 positivity (P = 0.06). No significant correlation between specific survival rate (determined by the log rank test) and the molecular markers analyzed was found, whereas the number of compromised lymph nodes and advanced TNM (tumor, node, metastasis) staging were associated with diminished survival.

Alguns aspectos do comportamento de cepas silvestres de Trypanosoma cruzi em camundongos e Calomys callosus (Rodentia)

Foram estudadas quatro cepas silvestres de T. cruzi: M226 isolada de Calomys callosus (Rodentia) e R52, R64 e R65 isoladas de Didelphis albiventris (Marsupialia). Estes animais foram coletados no município de Formosa, Goiás, Brasil. Os aspectos abordados, relacionados com o comportamento destas cepas em camundongos "swiss" 40 e C. callosus nascidos em laboratório, foram: parasitemia, prepatência, letalidade e histopatologia. Os resultados indicaram que as quatro cepas tinham baixa virulência para os animais testados. A parasitemia sempre se apresentou baixa e regular para os C. callosus. Nos camundongos, só raramente a parasitemia era patente. A prepatência nos C. callosus variou em média entre 9,2 a 10,2 dias, enquanto nos camundongos ficou entre 12-48 dias. A letalidade para C. callosus foi 7,7% e para camundongos 12,0%. Os estudos histopatológicos mostraram que somente 17,2% dos C. callosus apresentaram parasitismo tissular, enquanto em 25% dos camundongos foram encontrados pseudocistos íntegros ou rompidos. Houve um nítido miotropismo das quatro cepas tanto nos camundongos quanto nos C. callosus.

Epidemiological survey of Trypanosoma cruzi infection in North-Eastern Brazil using different diagnostic methods

A survey of the prevalence of Trypanosoma cruzi infection was carried out in Oitis, a small community in the State of Piaui, Brazil. Two hundred and sixty five individuals were screened by microscopic examination, hémoculture, indirect immunofluorescence (IFA), enzyme-linked immunosorbent assay (ELISA), and competitive enzyme-linked immunosorbent assay (C-ELISA) using the monoclonal antibody TCF87 against to a 25kd T. cruzi antigen. Seropositivity was 14.3% by the IFA test, 14.7% by ELISA, and 13.2% by C-ELISA. The C-ELISA using the TCF87 monoclonal antibody seems to be applicable in serodiagnosis of Chagas' disease.

Diagnosis of hepatitis C virus in Brazilian blood donors using a reverse transcriptase nested polymerase chain reaction: comparison with enzyme immunoassay and recombinant protein immunoblot assay

Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA > or = 3, ALT<1), group B (EIA > or = 3, ALT>1), group C (1<=EIA<3, ALT<1), group D (1<=EIA<3, ALT>1) and group E (EIA<=0.7). HCV sequences were detected by RT-nested PCR, using primers for the most conserved region of viral genome. RIBA-2 was applied to the same samples. In group A (n=6), all samples were positive by RT-nested PCR and RIBA-2. Among 124 samples in group B, 120 (96.8%) were RIBA-2 positive and 4 (3.2%) were RIBA-2 indeterminate but were seropositive for antigen c22.3. In group B, 109 (87.9%) of the RIBA-2 positive samples were also RT-nested PCR positive, as well as were all RIBA-2 indeterminate samples. In group C, all samples (n=9) were RT-nested PCR negative: 4 (44.4%) were also RIBA-2 negative, 4 (44.4%) were RIBA-2 positive and 1 (11.1%) was RIBA-2 indeterminate. HCV-RNA was detected by RT-nested PCR in 3 (37.5%) out of 8 samples in group D. Only one of them was also RIBA-2 positive, all the others were RIBA-2 indeterminate. All of the group E samples (controls) were RT- nested PCR and RIBA-2 negative. Our study suggests a strong relation between anti-HCV EIA-2 ratio > or = 3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR (75%), especially when the c22.3 band was detected.

Chagas disease: morbidity profile in an endemic area of Northeastern Brazil

Abstract: INTRODUCTION : This study evaluated the clinical forms and manifestation severities of Chagas disease among serologically reactive individuals from Western Rio Grande do Norte (Northeastern Brazil). METHODS : This cross-sectional study included 186 adults who were evaluated using electrocardiography, echocardiography, chest radiography, and contrast radiography of the esophagus and colon. A clinical-epidemiological questionnaire was also used. RESULTS : The indeterminate, cardiac, digestive, and cardiodigestive clinical forms of Chagas disease were diagnosed in 51.6% (96/186), 32.2% (60/186), 8.1% (15/186) and 8.1% (15/186) of the participants, respectively. Heart failure (functional classes I-IV) was detected in 7.5% (14/186) of the participants, and 36.4% (24/66), 30.3% (20/66), 15.2% (10/66), 13.6% (9/66), and 4.5% (3/66) of the patients were at stage A, B1, B2, C, and D, respectively. Dilated cardiomyopathy and electrocardiographic changes were detected in 10.2% (19/186) and 48.1% (91/186) of the participants, respectively. Apical aneurysm was diagnosed in 10.8% (20/186) of the participants, and other changes in the segmental myocardial contractility of the left ventricle were diagnosed in 33.9% (63/186) of the participants. Megaesophagus (groups I-IV) was observed in 7% (13/186) of the participants, megacolon (grades 1-3) was detected in12.9% (24/186) of the participants, and both organs were affected in 29.2% (7/24) of the megacolon cases. CONCLUSIONS : We detected various clinical forms of Chagas disease (including the digestive form). Our findings indicate that clinical symptoms alone may not be sufficient to exclude or confirm cardiac and/or digestive damage, and the number of patients with symptomatic clinical forms may be underestimated.

BIOLOGIA DE Nystalea nyseus (CRAMER, 1775) (LEPIDOPTERA: NOTODONTIDAE) EM FOLHAS de Eucalyptus urophylla

Essa pesquisa foi conduzida no laboratório de Entomologia Florestal da Universidade Federai de Viçosa à 25 ± 2" C, 60 ± 10% UR e fotoperiodo de 12L:12E. O objetivo foi estudar a biologia de Nystalea nyseus(Cramer, 1775) (Lepidoptera: Notodontidae) em folhas de Eucalyptus urophylla. 25;. nyseusapresentou cinco estádios, com duração, média, de 3,99 ± 0,11; 3,80 ±0,10; 4,95 ± 0,30; 5,96 ±0,21 e 6,85 ±0,14 dias, respectivamente. A duração total da fase larval foi de 25,5 dias. A fase de pré-pupa teve duração de 3,05 ± 0,05; 3,08 ± 0,06 dias e a de pupa 14,75 ± 0,45 c 13,82 ± 0,29 dias para machos e fêmeas, respectivamente. A viabilidade, dos ovos, foi de 74,72% e o período embrionário de 3,40 ± 0,16 dias. A longevidade de adultos, acasalados, foi de 6,71 ± 0,74 dias para machos e 9,14 ± 0,96 dias para fêmeas e razão sexual de 0,55, ou seja uma fêmea para 0,81 macho.

Avaliação dos efeitos a curto prazo de programas de conscientização e de terapia com pravastatina em indivíduos de alto risco para doença cardiovascular provenientes de consultórios particulares

OBJETIVO: Avaliar a eficácia de programas de conscientização (PC) sobre o controle de fatores de risco (FR) para doença cardiovascular (DCV). MÉTODOS: Pacientes hipercolesterolêmicos de alto risco para DCV foram divididos em 2 grupos durante 16 semanas. O grupo A (n=417, 54,3±10,0 anos, 55% homens) recebeu orientação verbal e escrita sobre controle de FR, e o grupo B (n=180, 54,4±10,9 anos, 45% homens) apenas orientação verbal. Todos os participantes receberam 10mg/dia de pravastatina por 12 semanas. Avaliaram-se o peso, pressão arterial, o colesterol total (CT) e frações, triglicérides, índices I e II de Castelli (CT/HDL-C e LDL-C/HDL-C) e escores de Framingham. RESULTADOS: No basal, A diferiu de B no HDL-C (40,0±11,0 vs 43,0±11,0mg/dl, p=0,013) e no índice I (8,2±3,0 vs 7,6±2,3, p=0,008). Após 16 semanas as variações % foram maiores em A do que B no CT (-28,0 vs -25,0, p<0,05), LDL-C (-29,0 vs -27,6, p<0,05), HDL-C (+13,7 vs. + 10,8, p<0,05) e índice I (-39,0 vs -33,0; p<0,05). A pravastatina potencializou os efeitos da dieta sobre os lípides. CONCLUSÃO: O PC parece ser mais eficaz a curto prazo, em reduzir os FR para DCV do que a orientação apenas verbal.

Correlation between turbidimetric and nephelometric methods of measuring C-reactive protein in patients with unstable angina or non-ST elevation acute myocardial infarction

OBJECTIVE: To evaluate the performance of the turbidimetric method of C-reactive protein (CRP) as a measure of low-grade inflammation in patients admitted with non-ST elevation acute coronary syndromes (ACS). METHODS: Serum samples obtained at hospital arrival from 68 patients (66±11 years, 40 men), admitted with unstable angina or non-ST elevation acute myocardial infarction were used to measure CRP by the methods of nephelometry and turbidimetry. RESULTS: The medians of C-reactive protein by the turbidimetric and nephelometric methods were 0.5 mg/dL and 0.47 mg/dL, respectively. A strong linear association existed between the 2 methods, according to the regression coefficient (b=0.75; 95% C.I.=0.70-0.80) and correlation coefficient (r=0.96; P<0.001). The mean difference between the nephelometric and turbidimetric CRP was 0.02 ± 0.91 mg/dL, and 100% agreement between the methods in the detection of high CRP was observed. CONCLUSION: In patients with non-ST elevation ACS, CRP values obtained by turbidimetry show a strong linear association with the method of nephelometry and perfect agreement in the detection of high CRP.

Disfunção diastólica em pacientes diabéticos normotensos, independente da presença de microangiopatia

OBJETIVO: Avaliar as alterações Doppler-ecocardiográficas de pacientes normotensos, com diabetes mellitus tipo II, na presença ou ausência de sinais de microangiopatia. MÉTODOS: Pacientes com diabetes mellitus tipo II foram submetidos a exames de fundo de olho contrastado com fluoresceína e dosagem de microalbuminúria para diagnóstico de microangiopatia, e divididos em dois grupos: DMII (pacientes sem microangiopatia, n=19) e DM+A (pacientes com microangiopatia, n=13). Todos foram submetidos a Doppler-ecocardiografia e os resultados comparados com normotensos de mesmo sexo e idade (grupo C, n=20), aplicando-se a ANOVA, seguida de teste de Tukey. Em todas as comparações, adotou-se como nível de significância p<0,05. RESULTADOS: Não houve diferenças entre os grupos quanto aos indicadores de função sistólica ou massa do ventrículo esquerdo. Foram observadas diferenças compatíveis com disfunção diastólica nos dois grupos de diabéticos, independente da presença de microangiopatia, que exibiram valores significativamente maiores dos tempos de relaxamento isovolumétrico do ventrículo esquerdo (TRIVE, ms): (DMII=97±22,2; DM+A=107±28,2 e C=80±10,7; p<0,05), e menores das velocidades máximas da onda de enchimento ventricular rápido (E, cm/s): (DMII=69±17,5; DM+A=75±19,7 e C=84±14,5, p<0,05 entre DMII e C). Sem diferença entre os grupos quanto à razão E/A. CONCLUSÃO: Pacientes normotensos com diabetes mellitus tipo II e sem sinais clínicos de comprometimento cardiovascular, apresentaram sinais de disfunção diastólica não associados à presença de microangiopatia.

Índice Tornozelo-Braquial (ITB) determinado por esfigmomanômetros oscilométricos automáticos

FUNDAMENTO: Índice Tornozelo-Braquial (ITB) é essencial na prática clínica, mas dificuldades técnicas na sua execução pelo padrão de referência Doppler vascular (DV) tornam-no ainda pouco utilizado. OBJETIVO: Avaliar aplicabilidade da determinação do ITB com uso de esfigmomanômetros oscilométricos automáticos (EOA) e sugerir a utilização dos índices delta-Bráquio-Braquial (delta-BB) e delta-ITB como marcadores de risco cardiovascular. MÉTODOS: Estudo descritivo e observacional de 247 pacientes ambulatoriais (56,2% feminino, média 62,0 anos) submetidos à determinação do ITB com aferição simultânea da pressão arterial (PA) em membros superiores (MMSS) e inferiores (MMII) utilizando-se dois EOA (OMRON-HEM705CP). Nos casos em que não foi possível aferir PA em pelo menos um dos MMII utilizou-se DV. Os pacientes divididos em Grupo N (ITB normal: 0,91 a 1,30) e Grupo A (ITB alterado: <0,90 ou >1,30) tiveram comparados entre si os valores de delta-ITB (diferença absoluta ITB/MMII) e delta-BB (diferença absoluta PAS/MMSS). RESULTADOS: Utilizando-se EOA foi possível determinar ITB em 90,7%. Com dados do Grupo N determinaram-se valores de referência (VR) no percentil 95 de delta-ITB (0-0,13) e delta-BB (0-8 mmHg). Quando comparado com o Grupo N, o Grupo A apresentou prevalência mais elevada tanto de delta-ITB (30/52 contra 10/195; Razão de Chances: 25,23; p<0,0001) como de delta-BB (13/52 contra 7/195; Razão de Chances: 8,95; p<0,0001) acima dos VR. CONCLUSÃO: O ITB pode ser determinado na maioria das vezes com EOA. Delta-ITB e delta-BB acima dos VR estiveram significativamente mais prevalentes nos portadores de ITB alterado e podem ser sugeridos como marcadores de risco cardiovascular em futuros estudos epidemiológicos.

Associação entre peso de nascimento e fatores de risco cardiovascular em adolescentes

FUNDAMENTO: Peso ao nascer (PN) é um determinante de risco a médio e longo prazo de fatores de risco cardiovascular. OBJETIVO: Estudar a associação entre peso ao nascer e fatores de risco cardiovascular em adolescentes de Salvador. MÉTODOS: Estudo de corte transversal com grupos de comparação por PN. Amostra composta de 250 adolescentes, classificados segundo IMC: normal alto (>p50 e 2.500g), peso normal (2.500g2%) meninas, idade 13,74 ± 2,03 anos, PN normal 80,8%, baixo PN 8,0% e alto PN 11,2%. Observou-se maior frequência de obesidade (42,9%, p=0,005), PAS e PAD elevadas (42,9%, p=0,000 e 35,7%, p=0,007, respectivamente) e síndrome metabólica (46,4%,p =0,002) no grupo com PN alto em relação ao PN normal. Indivíduos de alto PN apresentaram RP para PAS elevada 3,3(I.C. 95%, 1,7-6,4) e para obesidade 2,6 (I.C. 95%, 1,3-5,2) em relação aos com PN normal. A CA foi 83,3 ± 10,1 [p=0,038] nos adolescentes com alto PN. O perfil lipídico não mostrou diferenças estatisticamente significantes. CONCLUSÃO: Os dados sugerem que obesidade, PAS e PAD elevadas e síndrome metabólica na adolescência têm chance significativa de associar-se a alto peso no nascimento.

S&#237;ndrome Metab&#243;lica, Strain e Redu&#231;&#227;o da Fun&#231;&#227;o Mioc&#225;rdica: Multi-Ethnic Study of Atherosclerosis

Fundamento: A doen&#231;a cardiovascular subcl&#237;nica &#233; prevalente em pacientes com s&#237;ndrome metab&#243;lica (SM). O strain circunferencial (εCC) e o strain longitudinal (εLL) do ventriculo esquerdo (VE), avaliados pelo ecocardiograma com speckle tracking (STE), s&#227;o &#237;ndices de fun&#231;&#227;o sist&#243;lica: o encurtamento das fibras circunferenciais e longitudinais do VE &#233; indicado por um valor negativo do strain. Portanto, quanto mais negativo o strain, melhor a fun&#231;&#227;o sist&#243;lica do VE. O εCC e o εLL t&#234;m sido usados para demonstrar disfun&#231;&#227;o ventricular subcl&#237;nica em v&#225;rios dist&#250;rbios cl&#237;nicos. Objetivo: Levantamos a hip&#243;tese de que a SM est&#225; associada com comprometimento da fun&#231;&#227;o mioc&#225;rdica, quando avaliada pelo STE. M&#233;todos: Este estudo analisou participantes do Multi-Ethnic Study of Atherosclerosis (MESA) que realizaram o STE e foram avaliados para todos os componentes da SM. Resultados: Entre os 133 participantes inclu&#237;dos (mulheres: 63%; idade: 65 ± 9 anos), a preval&#234;ncia de SM foi de 31% (41/133). Indiv&#237;duos com SM apresentaram valores menores do εCC e do εLL que aqueles sem SM (-16,3% ± 3,5% vs. -18,4% ± 3,7%, p < 0,01; e -12,1% ± 2,5% vs. -13,9% ± 2,3%, p < 0,01, respectivamente). A fra&#231;&#227;o de eje&#231;&#227;o do VE (FEVE) foi semelhante nos dois grupos (p = 0,09). Na an&#225;lise multivariada, a SM associou-se a um valor mais baixo do strain circunferencial (B = 2,1%, IC 95%: 0,6-3,5; p < 0,01), mesmo ap&#243;s ajuste para idade, etnia, massa VE e FEVE. De maneira semelhante, a presen&#231;a de SM (B = 1,3%, IC 95%: 0,3-2,2; p < 0,01) e a massa do VE (B = 0,02%, IC 95%: 0,01 0,03; p = 0,02) associaram-se a um valor menor do strain longitudinal ap&#243;s ajuste para etnia, FEVE e creatinina. Conclus&#227;o: O εCC e o εLL do VE, marcadores de doen&#231;a cardiovascular subcl&#237;nica, est&#227;o comprometidos em indiv&#237;duos assintom&#225;ticos com SM e sem hist&#243;ria pr&#233;via de infarto mioc&#225;rdico, insufici&#234;ncia card&#237;aca e/ou de FEVE < 50%.