Recovery of DNA for the detection and genotyping of human papillomavirus from clinical cervical specimens stored for up to 2 years in a universal collection medium with denaturing reagent

The recovery and stability of DNA for the detection and genotyping of HPV in UCM-containing specimens, after exposure to denaturing reagents and stored for up to 2 years were evaluated. Samples were collected from 60 women who had cervical cytology specimens harboring cervical intraepithelial neoplasia (CIN) 2 or 3. All samples were stored in UCM and had been frozen at -20 degrees C following the addition of the denaturing reagent (sodium hydroxide) and the removal of the aliquot required for Hybrid Capture 2 testing for the identification of HPV DNA. The samples had been stored for 6, 12 and 24 months (20 samples for each storage time). HPV DNA extraction was performed according to a protocol designed specifically and the presence and quality of DNA was confirmed by human P-globin detection using the consensus primers G73 and G74. HPV DNA was amplified using the consensus primers PGMY09 and PGMY11, and reverse line-blot hybridization was used to detect type-specific amplicons for 37 HPV types. The DNA extracted from the denatured specimen was recovered in 57/60 (95%) of the samples. HPV DNA was detected in 56/57 (98%) of the recovered samples. Twenty-six of the 56 samples recovered (48%) were genotyped successfully. (c) 2007 Elsevier B.V. All rights reserved.

Association of dental enamel lead levels with risk factors for environmental exposure

OBJECTIVE: To analyze household risk factors associated with high lead levels in surface dental enamel. METHODS: A cross-sectional study was conducted with 160 Brazilian adolescents aged 14-18 years living in poor neighborhoods in the city of Bauru, southeastern Brazil, from August to December 2008. Body lead concentrations were assessed in surface dental enamel acid-etch microbiopsies. Dental enamel lead levels were measured by graphite furnace atomic absorption spectrometry and phosphorus levels were measured by inductively coupled plasma optical emission spectrometry. The parents answered a questionnaire about their children's potential early (05 years old) exposure to well-known lead sources. Logistic regression was used to identify associations between dental enamel lead levels and each environmental risk factor studied. Social and familial covariables were included in the models. RESULTS: The results suggest that the adolescents studied were exposed to lead sources during their first years of life. Risk factors associated with high dental enamel lead levels were living in or close to a contaminated area (OR = 4.49; 95% CI: 1.69;11.97); and member of the household worked in the manufacturing of paints, paint pigments, ceramics or batteries (OR = 3.43; 95% CI: 1.31;9.00). Home-based use of lead-glazed ceramics, low-quality pirated toys, anticorrosive paint on gates and/or sale of used car batteries (OR = 1.31; 95% CI: 0.56;3.03) and smoking (OR = 1.66; 95% CI: 0.52;5.28) were not found to be associated with high dental enamel lead levels. CONCLUSIONS: Surface dental enamel can be used as a marker of past environmental exposure to lead and lead concentrations detected are associated to well-known sources of lead contamination.

Insegurança alimentar, vínculo mãe-filho e desnutrição infantil em área de alta vulnerabilidade social

OBJETIVO: analisar a insegurança alimentar e o vínculo inadequado mãe-filho como dois potenciais determinantes da desnutrição em crianças de quatro a seis anos de idade. MÉTODOS: estudo de caso-controle desenvolvido em Escolas Municipais de Educação Infantil (EMEIs) no Jardim Jaqueline, área de alta vulnerabilidade social do município de São Paulo, Brasil. Foram aplicados a Escala Brasileira de Insegurança Alimentar e o Protocolo de Avaliação do Vínculo Mãe-filho, além de coletadas informações biológicas e socio-econômicas. Para verificação dos efeitos de cada variável independente e controle dos efeitos das demais variáveis incluídas no modelo, foi utilizado o modelo de regressão logística múltipla. RESULTADOS: verificou-se que tanto a insegurança alimentar familiar (OR=3,6) como o vínculo inadequado mãe-filho (OR=9,4) estiveram associados com a desnutrição infantil (p<0,05), mesmo após o controle para o peso ao nascimento da criança e idade, estado conjugal e trabalho maternos. CONCLUSÕES: tanto a insegurança alimentar familiar (OR=3,6) como o vínculo mãe-filho inadequado (OR=9,4) mostraram-se fatores determinantes da ocorrência da desnutrição na população estudada.

Interação entre consumo alimentar e polimorfismos da GSTM1 e GSTT1 no risco para o câncer de cabeça e pescoço: estudo caso-controle em São Paulo, Brasil

O objetivo foi investigar a interação entre fatores dietéticos e polimorfismos de enzimas de metabolização de xenobióticos (GSTM1 e GSTT1) associadas ao câncer de cabeça e pescoço em um estudo caso controle de base hospitalar, no Município de São Paulo, Brasil. Participaram 103 casos incidentes, histologicamente confirmados, e 101 controles. O consumo alimentar foi obtido por um questionário de frequência alimentar validado. Os polimorfismos GSTM1 e GSTT1 foram avaliados pelo método PCR. Observou-se aumento de risco no mais alto tercil de consumo de carne bovina na presença do alelo nulo da GSTM1 (OR = 10,79; IC95%: 2,17-53,64) e GSTT1 (OR = 3,41; IC95%: 0,43-27,21). Considerando-se a razão entre alimentos de origem animal e vegetal, verificou-se para o tercil intermediário a OR = 2,02 (IC95%: 0,24-16,0) e no tercil superior OR = 3,23 (IC95%: 0,40-25,92). Os resultados apontam para uma possível interação entre o consumo de carne e variantes polimórficas dos genes GSTM1 e GSTT1 na modulação do risco para o câncer de cabeça e pescoço, influenciados pelo consumo de alimentos de origem vegetal.

Prevalência de asma e fatores de risco em escolares da cidade de São Paulo

OBJETIVO: Analisar a prevalência de asma e possíveis fatores de risco associados. MÉTODOS: Estudo transversal, integrante do International Study of Asthma and Allergies in Childhood. Participaram 561 escolares de seis a sete anos de idade, provenientes de 35 escolas públicas da cidade de São Paulo, escolhidas por sorteio, em 2002. A amostra incluiu 168 asmáticos e 393 não asmáticos, que responderam questionário constituído por 33 questões referentes a dados pessoais, familiares e ambientais. A associação entre asma e fatores de risco foi avaliada pela análise de regressão logística, considerando-se nível de significância estatística de 5%. RESULTADOS: Entre os escolares, 31,2% referiam sibilos nos 12 meses anteriores à entrevista. Os fatores de risco significativamente associados à asma foram: sexo masculino (OR=2,4;IC 95%: 1,4;4,2), mãe fumante no primeiro ano de vida (OR=2,0; IC 95%: 1,1;3,8), presença de eczema em locais característicos (OR=3,0; IC 95%:1,2; 7,6) e rinoconjuntivite (OR=2,4;IC 95%: 1,2; 4,8). CONCLUSÕES: A prevalência de asma na região estudada foi elevada e os fatores de risco relacionados foram: sexo masculino, sintomas de rinoconjuntivite no último ano, mãe fumante no primeiro ano de vida e presença de eczema em locais característicos.

Prevalência de anticorpos anti-Chlamydophila spp. em propriedades rurais com histórico de aborto bovino no estado do Paraná

Chlamydophila abortus é o agente etiológico do aborto epizoótico bovino, cujas manifestações clínicas mais freqüentes são aborto, nascimento de bezerros prematuros e de animais fracos, natimorto e repetição de cio em intervalos irregulares. O objetivo deste trabalho foi estimar a prevalência de anticorpos anti-Chlamydophila spp. em fêmeas bovinas de propriedades rurais com histórico de aborto, selecionadas dentro do delineamento amostral do Plano Nacional de Controle e Erradicação da Brucelose e Tuberculose no estado do Paraná. Foram testadas pela prova de fixação de complemento 3.102 amostras de soro de fêmeas bovinas (idade > 24 meses), provenientes de 373 propriedades. Ao total, 44 (1,42%) animais foram positivos com títulos > 32. A prevalência de focos foi de 8,82% (6,15%-12,17%). Animais confinados ou semi-confinados (OR=3.339, P=0.004), propriedade com menos de 35 matrizes (OR=3.339, P=0.017), presença de produtos do aborto na pastagem (OR=2.372, P=0.037) e aluguel de pasto (OR=3.398, P=0.006) foram considerados fatores de risco para Chlamydophila spp. A infecção por Chlamydophila spp. acometeu um número pequeno de animais, oriundos de propriedades com histórico de aborto. A importância deste agente como causa de aborto em bovinos no estado do Paraná, se existir, é muito pequena.

Prevalência e fatores associados ao tabagismo em escolares da Região Sul do Brasil

OBJETIVO: Estimar a prevalência do tabagismo em estudantes e os fatores associados.MÉTODOS: Foram utilizados dados secundários, provenientes do inquérito Vigescola realizado em Curitiba (PR), Florianópolis (SC) e Porto Alegre (RS) em 2002 e 2004. A amostra compreendeu 3.690 escolares de 13 a 15 anos, cursando as sétima e oitava séries do ensino fundamental e primeira do ensino médio, em escolas públicas e privadas. Para a análise dos resultados foram estimadas proporções ponderadas, odds ratio (OR), e utilizada a técnica de regressão logística múltipla.RESULTADOS: As taxas de prevalência de tabagismo corresponderam a 10,7 por cento (IC 95 por cento: 10,2;11,3) em Florianópolis, 12,6 por cento (IC 95 por cento: 12,4;12,9) em Curitiba e 17,7 por cento (IC 95 por cento: 17,4;18,0) em Porto Alegre. Os fatores associados ao tabagismo em escolares em Curitiba foram: sexo feminino (OR=1,49), pai fumante (OR=1,59), amigos fumantes (OR=3,46), exposição à fumaça do tabaco fora de casa (OR=3,26) e possuir algum objeto com logotipo de marca de cigarro (OR=3,29). Em Florianópolis, as variáveis associadas ao tabagismo foram escolares do sexo feminino (OR=1,26), ter amigos fumantes (OR=9,31), exposição à fumaça do tabaco em casa (OR=2,03) e fora de casa (OR=1,45) e ter visto propaganda em cartazes (OR=1,82). Em Porto Alegre, as variáveis que estiveram associadas com o uso de tabaco pelos escolares foram sexo feminino (OR=1,57), idade entre 14 anos (OR=1,77) e 15 anos (OR=2,89), amigos fumantes (OR=9,12), exposição à fumaça do tabaco em casa (OR=1,87) e fora de casa (OR=1,77) e possuir algo com logotipo de marca de cigarro (OR=2,83).CONCLUSÕES: Há elevada prevalência de tabagismo entre escolares de 13 a 15 anos, cujos fatores significativamente associados comuns às três capitais são: ter amigos fumantes e estar exposto à fumaça ambiental fora de casa

Recovery and upgrading bovine rumen protein by extrusion: effect of lipid content on protein disulphide cross-linking, solubility and molecular weight

Bovine rumen protein with two levels of residual lipids (1.9 per cent or 3.8 per cent) was subjected to thermoplastic extrusion under different temperatures and moisture contents. Protein solubility in different buffers, disulphide cross-linking and molecular weight distribution were determined on the extrudates. After extrusion, samples with 1.9 per cent residual lipids content had a higher concentration of protein insoluble by undetermined forces, irrespective of feed moisture and processing temperature used. Lipid content of 3.8 per cent in the feed material resulted in more protein participating in the extrudate network through non-covalent interactions (hydrophobic and electrostatic) and disulphide bonds. A small dependency of the extrusion process on moisture and temperature and a marked dependency on lipid content, especially phospholipid, was observed, Electrophoresis under non-reducing conditions showed that protein extrusion with low feed moisture promoted high molecular breakdown inside the barrel, probably due to intense shear force, and further protein aggregation at the die end

Eventos de vida produtores de estresse e queixas de insônia entre auxiliares de enfermagem de um hospital universitário no Rio de Janeiro: estudo Pró-Saúde

OBJETIVO: Avaliar a associação entre eventos de vida produtores de estresse (EVPE) e queixas de insônia (QI). MÉTODOS: Foram analisados dados seccionais de 695 auxiliares de enfermagem de um hospital universitário, participantes do Estudo Pró-Saúde - coorte de funcionários de uma universidade no Rio de Janeiro. As informações foram obtidas através de um questionário multidimensional e autopreenchido, que avaliou a ocorrência de EVPE nos últimos 12 meses, variáveis socioeconômicas e demográficas e QI. As QI foram analisadas como desfecho politômico (frequente, ocasional, e ausente). Odds ratios brutos e ajustados foram calculados através de regressão logística multinomial. RESULTADOS: A prevalência total de QI foi de 45,8% (16,7% frequentes e 29,1% ocasionais). Após ajuste por sexo, idade, estado civil, renda familiar per capita e regime de trabalho, os EVPE associados com QI frequentes foram: "rompimento de relação amorosa" (OR = 3,32; IC95% 1,90 - 5,78), "ter tido problemas graves de saúde" (OR = 2,82; IC95% 1,73 - 4,58); "dificuldades financeiras graves" (OR = 2,38; IC95% 1,46 - 3,88), e "mudança forçada de moradia" (OR = 1,97; IC95% 1,02 - 3,79). Com relação às QI ocasionais, houve associação apenas com rompimento de relação amorosa (OR = 2,30; IC95% 1,42 - 3,74) e dificuldades financeiras graves (OR = 1,87; IC95% 1,27 - 2,75). CONCLUSÕES: Dada a responsabilidade com vidas humanas que os auxiliares de enfermagem assumem durante seu horário de trabalho, nossos achados podem contribuir para ações mais efetivas, por parte dos serviços de saúde ao trabalhador, para lidar com o estresse nessa categoria

Practice of walking and its association with perceived environment among elderly Brazilians living in a region of low socioeconomic level

Background: The aim of this study was analyze associations between the practice of walking and environmental perception among elderly Brazilians in a region of low socioeconomic level. Methods: A cross-sectional study was conducted among 385 elderly people aged 60 years and over. To evaluate walking, the International Physical Activity Questionnaire (IPAQ), long version (leisure and transport modules) was used. The environment was evaluated by means of the Neighborhood Environmental Walkability Scale (NEWS) (adapted Brazilian version). For the statistical analysis, multiple logistic regression models were created separately for men and women. The practice of at least 150 minutes a week of walking was the dependent variable, and the variables of environmental perception were the independent variables. All the models were controlled for schooling level and age. Results: The proportion of elderly people active in walking was 56.9% for the men and 26.4% for the women. The perception of the presence of soccer fields (OR = 4.12) and their proximity, within ten minutes' walk from home (OR = 3.43), were associated with the practice of walking among the men. The perception of the presence of public squares (OR = 4.70) and the proximity of primary healthcare units, within ten minutes' walk from home (OR = 3.71), were associated with the practice of walking among the women. An association with adequate perception of vehicle traffic remained at the threshold of significance for the women. Conclusion: Accessibility of leisure structures such as football fields and public squares and of health services such as primary healthcare units were important environmental variables associated with the practice of walking among elderly people living in a region of low socioeconomic level in Brazil. These variables need to be taken into consideration when aiming to promote the practice of walking among elderly people living in similar regions.

The impact of obstructive sleep apnea on metabolic and inflammatory markers in consecutive patients with metabolic syndrome

Background: Obstructive Sleep Apnea (OSA) is tightly linked to some components of Metabolic Syndrome (MetS). However, most of the evidence evaluated individual components of the MetS or patients with a diagnosis of OSA that were referred for sleep studies due to sleep complaints. Therefore, it is not clear whether OSA exacerbates the metabolic abnormalities in a representative sample of patients with MetS. Methodology/Principal Findings: We studied 152 consecutive patients (age 48 +/- 9 years, body mass index 32.3 +/- 3.4 Kg/m(2)) newly diagnosed with MetS (Adult Treatment Panel III). All participants underwent standard polysomnography irrespective of sleep complaints, and laboratory measurements (glucose, lipid profile, uric acid and C-reactive protein). The prevalence of OSA (apnea-hypopnea index >= 15 events per hour of sleep) was 60.5%. Patients with OSA exhibited significantly higher levels of blood pressure, glucose, triglycerides, cholesterol, LDL, cholesterol/HDL ratio, triglycerides/HDL ratio, uric acid and C-reactive protein than patients without OSA. OSA was independently associated with 2 MetS criteria: triglycerides: OR: 3.26 (1.47-7.21) and glucose: OR: 2.31 (1.12-4.80). OSA was also independently associated with increased cholesterol/HDL ratio: OR: 2.38 (1.08-5.24), uric acid: OR: 4.19 (1.70-10.35) and C-reactive protein: OR: 6.10 (2.64-14.11). Indices of sleep apnea severity, apnea-hypopnea index and minimum oxygen saturation, were independently associated with increased levels of triglycerides, glucose as well as cholesterol/HDL ratio, uric acid and C-reactive protein. Excessive daytime sleepiness had no effect on the metabolic and inflammatory parameters. Conclusions/Significance: Unrecognized OSA is common in consecutive patients with MetS. OSA may contribute to metabolic dysregulation and systemic inflammation in patients with MetS, regardless of symptoms of daytime sleepiness.

Pharmacogenetics of OATP Transporters Reveals That SLCO1B1 c.388A > G Variant Is Determinant of Increased Atorvastatin Response

Aims: The relationship between variants in SLCO1B1 and SLCO2B1 genes and lipid-lowering response to atorvastatin was investigated. Material and Methods: One-hundred-thirty-six unrelated individuals with hypercholesterolemia were selected and treated with atorvastatin (10 mg/day/4 weeks). They were genotyped with a panel of ancestry informative markers for individual African component of ancestry (ACA) estimation by SNaPshot (R) and SLCO1B1 (c.388A>G, c.463C>A and c.521T>C) and SLCO2B1 (-71T>C) gene polymorphisms were identified by TaqMan (R) Real-time PCR. Results: Subjects carrying SLCO1B1 c.388GG genotype exhibited significantly high low-density lipoprotein (LDL) cholesterol reduction relative to c.388AA+c.388AG carriers (41 vs. 37%, p = 0.034). Haplotype analysis revealed that homozygous of SLCO1B1*15 (c.521C and c.388G) variant had similar response to statin relative to heterozygous and non-carriers. A multivariate logistic regression analysis confirmed that c.388GG genotype was associated with higher LDL cholesterol reduction in the study population (OR: 3.2, CI95%: 1.3-8.0, p < 0.05). Conclusion: SLCO1B1 c.388A>G polymorphism causes significant increase in atorvastatin response and may be an important marker for predicting efficacy of lipid-lowering therapy.

Cell Therapy Attenuates Cardiac Dysfunction Post Myocardial Infarction: Effect of Timing, Routes of Injection and a Fibrin Scaffold

Background: Cell therapy approaches for biologic cardiac repair hold great promises, although basic fundamental issues remain poorly understood. In the present study we examined the effects of timing and routes of administration of bone marrow cells (BMC) post-myocardial infarction (MI) and the efficacy of an injectable biopolymer scaffold to improve cardiac cell retention and function. Methodology/Principal Findings: (99m)Tc-labeled BMC (6x10(6) cells) were injected by 4 different routes in adult rats: intravenous (IV), left ventricular cavity (LV), left ventricular cavity with temporal aorta occlusion (LV(+)) to mimic coronary injection, and intramyocardial (IM). The injections were performed 1, 2, 3, or 7 days post-MI and cell retention was estimated by gamma-emission counting of the organs excised 24 hs after cell injection. IM injection improved cell retention and attenuated cardiac dysfunction, whereas IV, LV or LV* routes were somewhat inefficient (< 1%). Cardiac BMC retention was not influenced by timing except for the IM injection that showed greater cell retention at 7 (16%) vs. 1, 2 or 3 (average of 7%) days post-MI. Cardiac cell retention was further improved by an injectable fibrin scaffold at day 3 post-MI (17 vs. 7%), even though morphometric and function parameters evaluated 4 weeks later displayed similar improvements. Conclusions/Significance: These results show that cells injected post-MI display comparable tissue distribution profile regardless of the route of injection and that there is no time effect for cardiac cell accumulation for injections performed 1 to 3 days post-MI. As expected the IM injection is the most efficient for cardiac cell retention, it can be further improved by co-injection with a fibrin scaffold and it significantly attenuates cardiac dysfunction evaluated 4 weeks post myocardial infarction. These pharmacokinetic data obtained under similar experimental conditions are essential for further development of these novel approaches.

Polymorphisms and DNA methylation of gene TP53 associated with extra-axial brain tumors

The p53 tumor suppressor gene is the most frequently mutated gene in human cancer; this gene is mutated in up to 50% of human tumors. It has a critical role in the cell cycle, apoptosis and cell senescence, and it participates in many crucial physiological and pathological processes. Polymorphisms of p53 have been suggested to be associated with genetically determined susceptibility in various types of cancer. Another process involved with the development and progression of tumors is DNA hypermethylation. Aberrant methylation of the promoter is an alternative epigenetic change in genetic mechanisms, leading to tumor suppressor gene inactivation. In the present study, we examined the TP53 Arg72Pro and Pro47Ser polymorphisms using PCR-RFLP and the pattern of methylation of the p53 gene by methylation-specific PCR in 90 extra-axial brain tumor samples. Patients who had the allele Pro of the TP53 Arg72Pro polymorphism had an increased risk of tumor development ( odds ratio, OR = 3.23; confidence interval at 95%, 95% CI = 1.71-6.08; P = 0.003), as did the allele Ser of TP53 Pro47Ser polymorphism (OR = 1.28; 95% CI = 0.03-2.10; P = 0.01). Comparison of overall survival of patients did not show significant differences. In the analysis of DNA methylation, we observed that 37.5% of meningiomas, 30% of schwannomas and 52.6% of metastases were hypermethylated, suggesting that methylation is important for tumor progression. We suggest that TP53 Pro47Ser and Arg72Pro polymorphisms and DNA hypermethylation are involved in susceptibility for developing extra-axial brain tumors.

VIABILITY OF SPORULATED OOCYSTS OF NEOSPORA CANINUM AFTER EXPOSURE TO DIFFERENT PHYSICAL AND CHEMICAL TREATMENTS

The aim of the present study was to evaluate the viability of Neospora caninum sporulated oocysts after various chemical and physical treatments. Bioassays in gerbils and molecular techniques (PCR-RFLP) were used for identification of the oocysts shed by experimentally infected dogs. Sporulated oocysts were purified and divided into 11 treatment groups as follows: absolute ethanol for 1 hr; 20 C for 6 hr; 4 C for 6 hr; 60 C for 1 min; 100 C for 1 min; 10% formaldehyde for 1 hr; 10% ammonia for 1 hr; 2% iodine for 1 hr; 10% sodium hypochlorite for I hr; 70% ethanol for I hr; and one group was left untreated and kept as a positive control. All chemical treatments were performed at room temperature (37 C). A total of 33 gerbils, or 3 gerbils per treatment, were used for bioassays. After treatment, the oocysts were divided into aliquots of 1,000 oocysts and orally administered to gerbils. After 63 days, the gerbils were anesthetized and killed with 0.2 ml of T61; blood and tissue samples were collected for serological (IFAT and western blotting), molecular (real-time PCR), histopathology, and immunohistochemical tests. Treatments were considered effective only if all 5 detection techniques tested negative. High temperatures at 100 C for 1 min and 10% sodium hypochlorite for 1 hr were the only treatments that met this condition, effectively inactivating all oocysts.