The HUman MicroNucleus project on eXfoLiated buccal cells (HUMNXL): The role of life-style, host factors, occupational exposures, health status, and assay protocol

The human buccal micronucleus cytome assay (BMCyt) is one of the most widely used techniques to measure genetic damage in human population studies. Reducing protocol variability, assessing the role of confounders, and estimating a range of reference values are research priorities that will be addressed by the HUMNXL, collaborative study. The HUMNXL, project evaluates the impact of host factors, occupation, life-style, disease status, and protocol features on the occurrence of MN in exfoliated buccal cells. In addition, the study will provide a range of reference values for all cytome endpoints. A database of 5424 subjects with buccal MN values obtained from 30 laboratories worldwide was compiled and analyzed to investigate the influence of several conditions affecting MN frequency. Random effects models were mostly used to investigate MN predictors. The estimated spontaneous MN frequency was 0.74 parts per thousand (95% CI 0.52-1.05). Only staining among technical features influenced MN frequency, with an abnormal increase for non-DNA-specific stains. No effect of gender was evident, while the trend for age was highly significant (p < 0.001). Most occupational exposures and a diagnosis of cancer significantly increased MN and other endpoints frequencies. MN frequency increased in heavy smoking (>= 40 cig/day. FR = 1.37:95% CI 1.03-.82) and decreased with daily fruit consumption (FR = 0.68; 95% CI 0.50-0.91). The results of the HUMNXL, project identified priorities for validation studies, increased the basic knowledge of the assay, and contributed to the creation of a laboratory network which in perspective may allow the evaluation of disease risk associated with MN frequency. (C) 2011 Elsevier B.V. All rights reserved.

Microdissecção e captura a laser na investigação do gene TP53 em tecidos incluídos em parafina

INTRODUÇÃO: Microdissecção e captura a laser (MCL) é uma técnica de desenvolvimento recente que permite a coleta de células individuais ou pequeno conjunto de células para análise molecular. Atualmente, no Brasil, há raros microscópios para MCL, de modo que a divulgação dos procedimentos inerentes a essa técnica é oportuna para destacar seu amplo potencial para diagnóstico e investigação. OBJETIVO: Este trabalho descreve a padronização dos procedimentos de MCL e de extração de DNA de material fixado em formalina e incluído em parafina. MATERIAL E MÉTODOS: Foram estudados o éxon 8 do gene TP53 e o gene da ciclofilina em amostras de tecido normal e de neoplasias de fígado e rim provenientes de modelo de carcinogênese química induzida em rato. A extração do DNA foi comprovada por reação em cadeia da polimerase (nested-PCR). RESULTADOS: Foram padronizados os procedimentos de preparo dos cortes histológicos, de microdissecção e captura a laser e de obtenção de seqüências gênicas pela reação de nested-PCR para tecidos incluídos em parafina. Obtivemos amplificação de 48,3% das amostras para o éxon 8 do gene TP53 e 51,7% para o gene da ciclofilina. Considerando pelo menos um dos dois segmentos gênicos, foram amplificadas 79,3% das amostras. DISCUSSÃO E CONCLUSÃO: A extração de DNA de tecidos fixados em formalina e incluídos em parafina e a técnica de nested-PCR foram adequadamente padronizadas para produtos gênicos de interesse, obtidos de material coletado por MCL. Esses procedimentos podem ser úteis para a obtenção de seqüências de DNA de arquivos para análise molecular.

Health surveys and use of maternal and child health care services in three municipalities within the São Paulo metropolitan area

Objectives. To describe the changes in the use of maternal and child health care services by residents of three municipalities-Embu, Itapecerica da Serra, and Taboao da Serra-in the São Paulo metropolitan area, 12 years after the implementation of the Unified Health System (SUS) in Brazil, and to analyze the potential of population-based health care surveys as sources of data to evaluate these changes.Methods. Two population-based, cross-sectional surveys were carried out in 1990 and 2002 in municipalities located within the São Paulo metropolitan area. For children under 1 year of age, the two periods were compared in terms of outpatient services utilization and hospital admission; for the mothers, the periods were compared in terms of prenatal care and deliveries. In both surveys, stratified and multiple-stage conglomerate sampling was employed, with standardization of interview questions.Results. The most important changes observed were regarding the location of services used for prenatal care, deliveries, and hospitalization of children less than 1 year of age. There was a significant increase in the use of services in the surrounding region or hometown, and decrease in the utilization of services in the city of São Paulo (in 1990, 80% of deliveries and almost all admissions for children less than 1 year versus 32% and 46%, respectively, in 2002). The use of primary care units and 24-hour walk-in clinics also increased. All these changes reflect care provided by public resources. In the private sector, there was a decrease in direct payments and payments through company-paid health insurance and an increase in payments through self-paid health insurance.Conclusions. The major changes observed in the second survey occurred simultaneous to the changes that resulted from the implementation of the SUS. Population-based health surveys are adequate for analyzing and comparing the utilization of health care services at different times.

Serological profile of buffalo (Bubalus bubalis) female calves vaccinated with standard Brucella abortus strain 19 vaccine using rose bengal, 2-mercaptoethanol and complement fixation tests

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo do leite de éguas sadias ou portadoras de mastite subclínica, pelo teste de Whiteside, análise microbiológica e contagem de células somáticas

Com o objetivo de identificar a microbiota existente no leite de éguas normais ou portadoras de mastite subclínica, coletaram-se amostras (10 a 15 ml) de leite de 38 animais, que foram examinadas pelo teste de Whiteside, após a homogeneização das amostras dos dois tetos de cada animal. Os resultados foram negativos. Os exames microbiológicos realizados em meios de ágar sangue bovino 10% e ágar MacConkey revelaram presença de bactérias do gênero Staphylococcus spp. e Streptococcus spp. A contagem de células somáticas de amostras individuais de cada teto revelou números superiores a 500.000 células/ml de leite em somente 9,3% das 73 amostras de leite examinadas. Os resultados sugerem a realização de novos estudos objetivando-se a padronização do número de células somáticas no leite de éguas.

Taenia saginata: polymerase chain reaction for taeniasis diagnosis in human fecal samples

The taeniasis-cysticercosis complex is a zoonosis of great medical and economic importance where humans play an important role as the carrier of adult stage of Taenia solium and Taenia saginata. This paper describes PCR standardization that can be applied in human fecal samples for taeniasis diagnosis. DNA extraction was achieved with DNAzol reagent, after egg disruption with glass beads. DNA prepared from fecal specimens was first purified and PCR amplified generating fragments of 170 and 600 bp. The assay described herein provides an important tool for T saginata identification in human fecal samples. (C) 2003 Elsevier B.V. (USA). All rights reserved.

Socio-behavioral factors influence prevalence and severity of dental caries in children with primary dentition

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histologic evaluation of the use of membrane, bone graft, and MTA in apical surgery

The aim of this study was to evaluate the periapical healing after the use of membrane, bone graft, and mineral trioxide aggregate (MTA) in apical surgery of dogs' teeth. Apical lesions were induced in 48 roots of 6 dogs after coronal access and pulpal removal. Apical surgery consisted of osteotomy with trephine bur for the standardization of the critical surgical cavities, followed by apicoectomy, curettage, preparation of the root-end cavities with the aid of the ultrasonic device, and retrofilling with MTA. The surgical sites were divided into: group 1-filled with blood; group 2-filled with blood and recovered with membrane; group 3-filled with bone graft; and group 4-filled with bone graft and recovered with membrane. The results showed that the inflammatory infiltrate, the periapical healing process, and the behavior of MTA was the same in all groups, including the mineralization stimulation. It was concluded that the use of membranes and bone graft materials isolated or associated in apical surgery did not alter the periapical healing process after the root-end filling with MTA. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 309-314)

Avaliação transversal e vertical da maxila, após expansão rápida, utilizando um método de padronização das radiografias póstero-anteriores

OBJETIVO: na presente pesquisa utilizou-se o aparelho Hyrax tendo como objetivo avaliar as alterações transversais e verticais da maxila. Foram selecionados 20 pacientes de ambos os gêneros, entre 7 e 11 anos de idade, que apresentavam mordida cruzada posterior uni ou bilateral. METODOLOGIA: foram realizadas telerradiografias em norma frontal (PA) antes e após a expansão. A fim de evitar que a falta de padronização dessas radiografias pudesse influenciar nos resultados, idealizou-se um método de padronização para tais radiografias. RESULTADOS E CONCLUSÕES: os resultados mostraram que a correção da mordida cruzada posterior com o aparelho Hyrax, que ocorreu em todos os casos, foi em função tanto da abertura da sutura palatina mediana como também da inclinação dos molares. A medida JgE-JgD relacionada com a expansão maxilar aumentou significantemente em todos os casos tratados. Nas avaliações dentárias, as medidas verticais não apresentaram diferenças significantes, ao contrário das medidas transversais (FTE-FTD; FBE-FBD; LO.BTE; LO.BTD; BTE.BTD). A largura da cavidade nasal também aumentou significantemente (CNE-CND). O método de padronização proposto permite, portanto, utilizar as medidas que avaliam a expansão rápida da maxila, através das radiografias póstero-anteriores, com segurança, também no sentido vertical.

Exploring CBCT-Based DICOM Files. A Systematic Review on the Properties of Images Used to Evaluate Maxillofacial Bone Grafts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of the radiopacity of calcium hydroxide- and glass-ionomer-based root canal sealers

Aim To evaluate the radiopacity of calcium hydroxide-based root canal sealers (Acroseal, Sealapex and Sealer 26), a glass-ionomer-based sealer (Activ GP Sealer) and a zinc oxide and eugenol-based sealer (Intrafill).Methodology Five disc-shaped specimens (10 x 1 mm) were fabricated from each material, according to the International Organization for Standardization (ISO) 6876/2001 standard. After setting of the materials, radiographs were taken using occlusal films and a graduated aluminum step-wedge varying from 2 to 16 mm in thickness. The dental X-ray unit (GE1000) was set at 50 Kvp, 10 mA, 18 pulses s(-1) and distance of 33.5 cm. The radiographs were digitized and the radiopacity compared with that of the aluminum step-wedge, using WIXWIN-2000 software (Gendex). Data (mm Al) were submitted to ANOVA and Tukey test.Results Intrafill was the most radiopaque material (7.67 mm Al) followed by Sealer 26 (6.33 mm Al), Sealapex (6.05 mm Al) and Acroseal (4.03 mm Al). Activ GP was the least radiopaque material (1.95 mm Al, P < 0.05).Conclusions The sealers evaluated in this study had different radiopacities. However, except for the glass-ionomer-based sealer, all materials had radiopacity values above the minimum recommended by the ISO standard.

Radiopacity evaluation of new root canal filling materials by digitalization of images

The aim of this study was to evaluate the radiopacity of five root canal filling materials (AH Plus, Intrafill, Roeko Seal, Epiphany, and EndoRez). Following the International Organization of Standardization 687612001, five circular specimens (10 X 1 mm) were made from each material. After the material set, radiographs were made using occlusal film and a graduated aluminum step-wedge varying in thickness from 2 to 16 mm. The dental X-ray unit (GE1000) was set at 50 Kvp, 10 mA, 18 pulses/second, and distance of 33.5 cm. The radiographs were digitized, and the radiopacity was compared with the aluminum step-wedge, using WIXWIN-2000 software (Gendex). Data (mm Al) were analyzed using ANOVA and Tukey tests. AH Plus and Epiphany were the most radiopaque materials (9.8 and 8.8 mm Al, respectively), followed by EndoRez (7.2 mm Al). Roeko Seal and Intrafill presented the lowest radiopacity values (5.7 and 6.1 mm Al, respectively). Although the materials evaluated demonstrated different radiopacities, all had values above the minimum recommended by the international Organization of Standardization.

Radiographic effect of different radiopacifiers on a potential retrograde filling material

Objective. The objective of this study was to evaluate the radiopacity of white Portland cement (PC) associated with bismuth oxide (Bi(2)O(3)), barium sulfate (BaSO(4)), iodoform (CHI(3)), and zirconium oxide (ZrO(2)). White mineral trioxide aggregate (WMTA) and PC without radiopacifier were used as positive and negative controls, respectively.Study design. Following International Organization for Standardization (ISO) 6876/2001, 5 circular specimens (10 x 1 mm) were made from each material. After the materials set, radiographs were taken using occlusal film and a graduated aluminum step-wedge varying in thickness from 2 to 16 mm. The dental X-ray unit (GE1000) was set at 50 Kvp, 10 mA, 18 pulses/s, and distance of 33.5 cm. The radiographs were digitized, and radiopacity was compared with the aluminum step-wedge, using Wixwin-2000 software (Gendex). Data (mm Al) were analyzed using analysis of variance and Tukey tests.Results. The PC + Bi(2)O(3) and WMTA samples presented greater radiopacity (5.88 and 5.72 mm Al, respectively), followed by PC + ZrO(2) (3.87 mm Al) and PC + CHI(3) (3.50 mm Al). The PC + BaSO(4) and PC samples presented the lowest radiopacity values (2.35 and 1.69 mm Al, respectively), which were below the minimum value recommended by the ISO.Conclusion. Analysis of the present results led to the conclusion that all of the materials tested presented acceptable radiopacity, except PC + BaSO(4) and pure PC. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 108: 628-632)

Early juvenile development of Armases rubripes (Rathbun 1897) (Crustacea, Brachyura, Sesarmidae) and comments on the morphology of the megalopa and first crab

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of lymphocyte cultures for BrdU replication banding patterns in anuran species (Amphibia)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)