Recent evidence suggests that oxidatively modified forms of low-density lipoprotein (LDL) may be particularly atherogenic. In this investigation, the catabolism of human LDL modified by lipid peroxidation in vitro was studied with a recirculating rat liver perfusion system. A dual-labelling technique was used that permitted native LDL and modified LDL to be studied simultaneously in the liver perfusion system. Native human LDL was found to have a fractional catabolic rate (FCR) of 1.00 +/- 0.21%/h, in agreement with other investigators. Subjecting LDL to oxidation for 12 h in the presence of 30 microM FeEDTA did not significantly affect its FCR. LDL treated with a superoxide-generating system (xanthine oxidase, hypoxanthine, O2) in the presence of 30 microM FeEDTA did, however, show a significant increase in FCR (3.23 +/- 0.19%/h). The hepatic uptakes of native LDL and LDL oxidized with FeEDTA+O2 were similar, but both were significantly lower than the hepatic uptake of LDL treated with the superoxide-radical-generating system. The proteolysis of LDL with pancreatin did not influence either its susceptibility to oxidation or its FCR. LDL oxidation resulted in the preferential loss of alpha-tocopherol rather than gamma-tocopherol. These data indicate that the rat liver effectively catabolizes LDL oxidatively modified by treatment with the superoxide-generating system. Furthermore, our results suggest that only very low plasma levels of highly oxidized LDL could be found under conditions in vivo. The liver may therefore play a major role in protecting the arterial vasculature from highly atherogenic forms of LDL.

INTRODUÇÃO: A poliquimioterapia (PQT-OMS) para tratamento da hanseníase resultou em drástica redução da sua prevalência, mas em limitado impacto na detecção de casos novos (CN), que se manteve estável em Santa Catarina, estado na fase de pós-eliminação. Casos com altos índices baciloscópicos apresentam risco de recidiva tardia e podem consistir em focos persistentes de transmissão da doença. OBJETIVOS: Avaliar a recidiva da doença em amostra de pacientes virchowianos regularmente tratados com PQT-OMS 24 doses; ensaios de resistência terapêutica em camundongos para os casos suspeitos; resposta imune específica (celular e humoral) e detecção do DNA do M. leprae nos casos-índices (CI) e seus contatos indradomiciliares (CID); e os achados frente aos indicadores epidemiológicos e operacionais de Santa Catarina. CASUÍSTICA E MÉTODOS: A partir da busca no Sistema de Informação de Agravos de Notificação (SINAN), foram selecionados 46 CI, tratados entre 1990-2000, e 187 CID, dos municípios de Itajaí e Joinville. A avaliação constituiu de: exames dermatoneurológico e anatomopatológico da pele; baciloscopia do esfregaço cutâneo; reação de Mitsuda; sorologia para glicolipídeo fenólico-I (IgM-anti-PGL-I); ensaios de resistência terapêutica em camundongos e de detecção do DNA bacilar no muco nasal por reação em cadeia da polimerase (PCR) para as seqüências repetitivas específicas RLEP-130 e RLEP-372. RESULTADOS: Entre os CI após alta por cura (m=11,2 ± 3 anos), a idade média (57,3±14,5 anos) foi superior (p<0,05) comparada aos CID, com predomínio de homens (p=0,001) e Mitsuda-negativos (78,6%). Em quatro casos (8,7%) considerados recidivados, o valor sérico médio (0,365) e as freqüências da positividade do anti-PGL-I e a da RLEP-130 (75%; p=0,03) foram consistentemente elevados, e os testes em camundongos, negativos. Dentre os 187 CID, 22 (11,8%) adoeceram (CIDd), a maioria (10 casos; 45,4%) foi diagnosticada entre 2 a 19 anos; 6 casos (27,3%), no mesmo período do seu respectivo CI; e 6 CN (3 dimorfo-tuberculóides e 3 tuberculóides) foram detectados durante a intervenção. Os CN evidenciaram elevada freqüência da positividade da RLEP-130 (50%) contrastante com a do anti-PGL-I (20%) e seu valor sérico médio (0,075) inferior. Entre os CI após a alta PQT-MB (>10 anos), houve inferioridade da freqüência da positividade (14,3%) e do valor sérico médio do anti-PGL-I (0,072), estabelecendo uma correlação negativa (p=0,038,r=-0,32) com o tempo decorrido. Valores similares da freqüência de positividade para RLEP-130 foram encontrados em: CI com alta por cura (26,7%), em períodos superiores (25%) a 10 anos de conclusão do tratamento e CID saudáveis (20,5%). A positividade para a RLEP-130 foi mais freqüente (p<0,001) comparada à da RLEP-372. CONCLUSÕES: A taxa de recidiva após PQT-MB 24 doses é baixa e prevalece alta percentagem de cura (91,3%). É racional considerar que o anti-PGL-I e a detecção do DNA bacilar por RLEP-130 devam ser analisados em conjunto com os demais achados clínicos e laboratoriais, ainda que nossos resultados possam ter diferenciado os grupos nas distintas condições: recidiva e alta por cura, e identificado doentes, contatos e famílias em risco. O seguimento e o monitoramento clínico e laboratorial por períodos prolongados incrementariam a detecção precoce de novos casos entre os comunicantes, desde que o intervalo para o diagnóstico foi longo para a maioria daqueles que adoeceram. Estas estratégias seriam potencialmente facilitadas em áreas com reduzida prevalência, e particularmente valiosas para a manutenção do controle na fase de pós-eliminação / Introduction: Multidrugtherapy (MDT-WHO) resulted in marked reduction of leprosy prevalence in Brazil, without impact on detection of new cases in Santa Catarina (SC) state in the post-elimination phase. Lepromatous cases with high bacilloscopic index have late relapse risk and can consist in the disease transmission focus. Objectives: The aim of this study was to evaluate the disease recurrence and microbiological resistance in lepromatous leprosy patients regularly treated with MDT-WHO/24 doses; specific immune response (cellular and humoral) and M. leprae DNA detection in index cases (IC) and their household contacts (HHC); and the findings face to the epidemiological and operational indicators of Santa Catarina state. Casuistic and methods: After consulting the Brazilian Information System of Notification (SINAN) database, 46 IC successfully diagnosed and treated between 1990 and 2000, and their 187 HHC from Joinville and Itajaí (SC) municipalities were selected. A dermatoneurological examination was performed, as well as the skin biopsies for histopathology and therapy resistance assay in mouse pads, skin smears for bacilloscopy, anti-PGL-I IgM serology, Mitsuda reaction, polymerase chain reaction for M. leprae DNA detection in nasal secretion based on 130bp and 372bp specific repetitive sequences (RLEP). Results: Cured IC (m=11.2±3 years) had mean age higher ((57.3±14.5 years old; p<0.05) than HHC, most of them were males (p=0.001) and Mitsuda negative (78.6%). In 4 relapsed cases (8.7%) the average anti-PGL-I serum levels (0.365), as well as frequency of positive antibody and RLEP-130 (75%; p=0.03) were consistently high and the mouse footpads assays resulted negative. Among 187 HHC, 22 (11.8%) became sick (sHHC), 10 cases (45.4%) were diagnosed between 2 and 19 years, being 6 cases (27.3%) in the same year of their IC; 6 new cases (3 borderline-tuberculoid and 3 tuberculoid) were detected during the study, with high RLEP-130 positive frequency (50%), as opposed to anti-PGL-I (20%) and low average serum levels (0.075). Among IC with more than 10 years of discharge, frequency of positive anti-PGL-I (14.3%) and average serum levels (0.072) were lower and had negative correlation (p=0.038, r= -0.32) with time after cure. Similar values of frequency of positivity for RLEP-130 were found: IC with discharge by cure (26.7%), when time interval was higher than 10 years (25%) and in healthy HHC (20.5%). RLEP-130 positivity was more frequent (p<0.001) than RLEP-372. Conclusions: Relapse rate after MDT-WHO 24 doses was low, with high cure rate (91.3%). Serology anti-PGL-I and M. leprae DNA detection by RLEP-130 must be analyzed together with other clinical and laboratory findings, though our results had differentiated groups in the following conditions: relapse and discharge by cure, patient identification, HHC and families at risk. Long term follow-up with laboratorial and clinical monitoring could lead to early detection of new cases among HHC, since the period between diagnoses was long for most sHHC. This strategy may be useful in areas with decreased prevalence, and of particular value for maintenance of disease control in the post-elimination phase

S.aureus resistente à meticilina(SARM) é um problema em centros de fibrose cística(FC). Foi desenvolvido um multiplex PCR(mPCR) para detecção do SARM em secreção respiratória de 106 pacientes com FC. Foram usados 3 pares de primers para amplificar os genes: mecA, coa, 16S rRNA. O mPCR detectou até 0,25pg de DNA de SARM e identificou 70/106(66,0%) pacientes com S.aureus e 28/106(26,4%) com SARM. O mPCR mostrou especificidade, sensibilidade, valores preditivos positivo e negativo de 87,8%, 84,4%, 50% e 97,5%, considerando a cultura como padrão-ouro. Os resultados discordantes foram testados com outros primers, confirmando os obtidos pelo mPCR em 82/84. O mPCR mostrou-se método rápido e confiável para detecção de SARM / Methicillin-resistant S.aureus(MRSA) is a significant concern in cystic fibrosis(CF) centers. A multiplex PCR(MPCR) was developed to detect MRSA in respiratory secretion of 106 CF patients. Three pairs of primers were used for amplification of genes: mecA, coa, 16S rRNA. MPCR detected 0.25pg of MRSA DNA and identified 70/106(66.0%) of patients with S.aureus and 28/106(26.4%) with MRSA. MPCR showed specificity, sensitivity, positive and negative predicted values at 87.8%, 84.4%, 50% and 97.5%, considering culture as the gold standard. Discrepant results were retested using different primers, and confirmed MPCR results in 82/84. The developed MPCR was found to be a rapid and reliable method for MRSA detection

Foi realizado um estudo para verificar a ocorrência de leveduras do gênero Rhodotorula, em hemocultura por um período de 8 anos. Os pacientes identificados foram descritos clinicamente segundo variáveis de interesse incluindo dados sobre terapêutica e desfecho. Determinou-se também as concentrações inibitórias mínimas de 20 cepas frente a diferentes antifúngicos de acordo com NCCLS e EUCAST. Realizou-se tipagem molecular através da cariotipagem eletroforética em campo pulsátil / A study was conducted to verify the frequency of occurrence of Rhodotorula spp. from blood cultures over an 8-year period, clinically and microbiologically characterizing patients affected, including data regarding antifungal treatment and outcome. The minimal inhibitory concentrations of antifungal agents were determined against 20 isolates. Molecular typing of the strains were performed using pulsed field gel electrophoresis method

S.aureus resistente à meticilina(SARM) é um problema em centros de fibrose cística(FC). Foi desenvolvido um multiplex PCR(mPCR) para detecção do SARM em secreção respiratória de 106 pacientes com FC. Foram usados 3 pares de primers para amplificar os genes: mecA, coa, 16S rRNA. O mPCR detectou até 0,25pg de DNA de SARM e identificou 70/106(66,0%) pacientes com S.aureus e 28/106(26,4%) com SARM. O mPCR mostrou especificidade, sensibilidade, valores preditivos positivo e negativo de 87,8%, 84,4%, 50% e 97,5%, considerando a cultura como padrão-ouro. Os resultados discordantes foram testados com outros primers, confirmando os obtidos pelo mPCR em 82/84. O mPCR mostrou-se método rápido e confiável para detecção de SARM / Methicillin-resistant S.aureus(MRSA) is a significant concern in cystic fibrosis(CF) centers. A multiplex PCR(MPCR) was developed to detect MRSA in respiratory secretion of 106 CF patients. Three pairs of primers were used for amplification of genes: mecA, coa, 16S rRNA. MPCR detected 0.25pg of MRSA DNA and identified 70/106(66.0%) of patients with S.aureus and 28/106(26.4%) with MRSA. MPCR showed specificity, sensitivity, positive and negative predicted values at 87.8%, 84.4%, 50% and 97.5%, considering culture as the gold standard. Discrepant results were retested using different primers, and confirmed MPCR results in 82/84. The developed MPCR was found to be a rapid and reliable method for MRSA detection

INTRODUÇÃO: A poliquimioterapia (PQT-OMS) para tratamento da hanseníase resultou em drástica redução da sua prevalência, mas em limitado impacto na detecção de casos novos (CN), que se manteve estável em Santa Catarina, estado na fase de pós-eliminação. Casos com altos índices baciloscópicos apresentam risco de recidiva tardia e podem consistir em focos persistentes de transmissão da doença. OBJETIVOS: Avaliar a recidiva da doença em amostra de pacientes virchowianos regularmente tratados com PQT-OMS 24 doses; ensaios de resistência terapêutica em camundongos para os casos suspeitos; resposta imune específica (celular e humoral) e detecção do DNA do M. leprae nos casos-índices (CI) e seus contatos indradomiciliares (CID); e os achados frente aos indicadores epidemiológicos e operacionais de Santa Catarina. CASUÍSTICA E MÉTODOS: A partir da busca no Sistema de Informação de Agravos de Notificação (SINAN), foram selecionados 46 CI, tratados entre 1990-2000, e 187 CID, dos municípios de Itajaí e Joinville. A avaliação constituiu de: exames dermatoneurológico e anatomopatológico da pele; baciloscopia do esfregaço cutâneo; reação de Mitsuda; sorologia para glicolipídeo fenólico-I (IgM-anti-PGL-I); ensaios de resistência terapêutica em camundongos e de detecção do DNA bacilar no muco nasal por reação em cadeia da polimerase (PCR) para as seqüências repetitivas específicas RLEP-130 e RLEP-372. RESULTADOS: Entre os CI após alta por cura (m=11,2 ± 3 anos), a idade média (57,3±14,5 anos) foi superior (p<0,05) comparada aos CID, com predomínio de homens (p=0,001) e Mitsuda-negativos (78,6%). Em quatro casos (8,7%) considerados recidivados, o valor sérico médio (0,365) e as freqüências da positividade do anti-PGL-I e a da RLEP-130 (75%; p=0,03) foram consistentemente elevados, e os testes em camundongos, negativos. Dentre os 187 CID, 22 (11,8%) adoeceram (CIDd), a maioria (10 casos; 45,4%) foi diagnosticada entre 2 a 19 anos; 6 casos (27,3%), no mesmo período do seu respectivo CI; e 6 CN (3 dimorfo-tuberculóides e 3 tuberculóides) foram detectados durante a intervenção. Os CN evidenciaram elevada freqüência da positividade da RLEP-130 (50%) contrastante com a do anti-PGL-I (20%) e seu valor sérico médio (0,075) inferior. Entre os CI após a alta PQT-MB (>10 anos), houve inferioridade da freqüência da positividade (14,3%) e do valor sérico médio do anti-PGL-I (0,072), estabelecendo uma correlação negativa (p=0,038,r=-0,32) com o tempo decorrido. Valores similares da freqüência de positividade para RLEP-130 foram encontrados em: CI com alta por cura (26,7%), em períodos superiores (25%) a 10 anos de conclusão do tratamento e CID saudáveis (20,5%). A positividade para a RLEP-130 foi mais freqüente (p<0,001) comparada à da RLEP-372. CONCLUSÕES: A taxa de recidiva após PQT-MB 24 doses é baixa e prevalece alta percentagem de cura (91,3%). É racional considerar que o anti-PGL-I e a detecção do DNA bacilar por RLEP-130 devam ser analisados em conjunto com os demais achados clínicos e laboratoriais, ainda que nossos resultados possam ter diferenciado os grupos nas distintas condições: recidiva e alta por cura, e identificado doentes, contatos e famílias em risco. O seguimento e o monitoramento clínico e laboratorial por períodos prolongados incrementariam a detecção precoce de novos casos entre os comunicantes, desde que o intervalo para o diagnóstico foi longo para a maioria daqueles que adoeceram. Estas estratégias seriam potencialmente facilitadas em áreas com reduzida prevalência, e particularmente valiosas para a manutenção do controle na fase de pós-eliminação / Introduction: Multidrugtherapy (MDT-WHO) resulted in marked reduction of leprosy prevalence in Brazil, without impact on detection of new cases in Santa Catarina (SC) state in the post-elimination phase. Lepromatous cases with high bacilloscopic index have late relapse risk and can consist in the disease transmission focus. Objectives: The aim of this study was to evaluate the disease recurrence and microbiological resistance in lepromatous leprosy patients regularly treated with MDT-WHO/24 doses; specific immune response (cellular and humoral) and M. leprae DNA detection in index cases (IC) and their household contacts (HHC); and the findings face to the epidemiological and operational indicators of Santa Catarina state. Casuistic and methods: After consulting the Brazilian Information System of Notification (SINAN) database, 46 IC successfully diagnosed and treated between 1990 and 2000, and their 187 HHC from Joinville and Itajaí (SC) municipalities were selected. A dermatoneurological examination was performed, as well as the skin biopsies for histopathology and therapy resistance assay in mouse pads, skin smears for bacilloscopy, anti-PGL-I IgM serology, Mitsuda reaction, polymerase chain reaction for M. leprae DNA detection in nasal secretion based on 130bp and 372bp specific repetitive sequences (RLEP). Results: Cured IC (m=11.2±3 years) had mean age higher ((57.3±14.5 years old; p<0.05) than HHC, most of them were males (p=0.001) and Mitsuda negative (78.6%). In 4 relapsed cases (8.7%) the average anti-PGL-I serum levels (0.365), as well as frequency of positive antibody and RLEP-130 (75%; p=0.03) were consistently high and the mouse footpads assays resulted negative. Among 187 HHC, 22 (11.8%) became sick (sHHC), 10 cases (45.4%) were diagnosed between 2 and 19 years, being 6 cases (27.3%) in the same year of their IC; 6 new cases (3 borderline-tuberculoid and 3 tuberculoid) were detected during the study, with high RLEP-130 positive frequency (50%), as opposed to anti-PGL-I (20%) and low average serum levels (0.075). Among IC with more than 10 years of discharge, frequency of positive anti-PGL-I (14.3%) and average serum levels (0.072) were lower and had negative correlation (p=0.038, r= -0.32) with time after cure. Similar values of frequency of positivity for RLEP-130 were found: IC with discharge by cure (26.7%), when time interval was higher than 10 years (25%) and in healthy HHC (20.5%). RLEP-130 positivity was more frequent (p<0.001) than RLEP-372. Conclusions: Relapse rate after MDT-WHO 24 doses was low, with high cure rate (91.3%). Serology anti-PGL-I and M. leprae DNA detection by RLEP-130 must be analyzed together with other clinical and laboratory findings, though our results had differentiated groups in the following conditions: relapse and discharge by cure, patient identification, HHC and families at risk. Long term follow-up with laboratorial and clinical monitoring could lead to early detection of new cases among HHC, since the period between diagnoses was long for most sHHC. This strategy may be useful in areas with decreased prevalence, and of particular value for maintenance of disease control in the post-elimination phase

Foi realizado um estudo para verificar a ocorrência de leveduras do gênero Rhodotorula, em hemocultura por um período de 8 anos. Os pacientes identificados foram descritos clinicamente segundo variáveis de interesse incluindo dados sobre terapêutica e desfecho. Determinou-se também as concentrações inibitórias mínimas de 20 cepas frente a diferentes antifúngicos de acordo com NCCLS e EUCAST. Realizou-se tipagem molecular através da cariotipagem eletroforética em campo pulsátil / A study was conducted to verify the frequency of occurrence of Rhodotorula spp. from blood cultures over an 8-year period, clinically and microbiologically characterizing patients affected, including data regarding antifungal treatment and outcome. The minimal inhibitory concentrations of antifungal agents were determined against 20 isolates. Molecular typing of the strains were performed using pulsed field gel electrophoresis method

We have compared two gastric string tests for obtaining gastric juice for culture of Helicobacter pylori and for nested-PCR detection and PCR-based combined single- and double-stranded conformational polymorphism (SDSCP) discrimination of infecting strains. String test specimens were obtained from one seropositive volunteer for 13 consecutive weeks. The distal 10 cm of each string was suspended in 1 ml saline and quantitatively cultured. An additional nine volunteers with histories of upper-gastrointestinal complaints were given a string test for culture and nested-PCR assay. H. pylori isolates and/or gastric juice from each volunteer were extracted for DNA and analyzed by PCR-based SDSCP. Quantitative culture showed that the Entero-test was four times as sensitive as the Gastro-test but was more prone to contamination by oral flora. However, the two string tests are equally sensitive by PCR assays. Thus, the Gastro-test is more suitable for culture detection of H. pylori, since it is less prone to oral contamination and its shorter length is better tolerated. SDSCP analysis of H. pylori DNA from four PCR-positive volunteers without requiring culture showed four distinct profiles, indicating different infecting strains. SDSCP analysis of strains isolated before and after treatment of one volunteer had the same SDSCP profile, suggesting endogenous reinfection by the same strain.

The nuclear lamina protein, lamin A is produced by proteolytic cleavage of a 74 kDa precursor protein, prelamin A. The conversion of this precursor to mature lamin A is mediated by a specific endoprotease, prelamin A endoprotease. Subnuclear fractionation indicates that the prelamin A endoprotease is localized at the nuclear membrane. The enzyme appears to be an integral membrane protein, as it can only be removed from the nuclear envelope with detergent. It is effectively solubilized by the detergent n-octyl-beta-D-glucopyranoside and can be partially-purified (approximately 1200-fold) by size exclusion and cation exchange (Mono S) chromatography. Prelamin A endoprotease from HeLa cells was eluted from Mono S with 0.3 M sodium chloride as a single peak of activity. SDS-PAGE analysis of this prelamin A endoprotease preparation shows that it contains one major polypeptide at 65 kDa and smaller amounts of a second 68 kDa polypeptide. Inhibition of the enzyme activity in this preparation by specific serine protease inhibitors is consistent with the enzyme being a serine protease.

Soil aquifer treatment was simulated in 1 m laboratory soil columns containing silica sand under saturated and unsaturated soil conditions to examine the effect of travel length through the unsaturated zone on the removal of wastewater organic matter, the effect of soil type on dissolved organic carbon removal and also the type of microorganisms involved in the removal process. Dissolved organic carbon removal and nitrification did enhance when the wastewater travelled a longer length through the unsaturated zone. A similar consortium of microorganisms was found to exist in both saturated and unsaturated columns. Microbial concentrations however were lowest in the soil column containing silt and clay in addition to silica sand. The presence of silt and clay was detrimental to DOC removal efficiency under saturated soil conditions due to their negative effect on the hydraulic performance of the soil column and microbial growth.