Comparative study of three different procedures for RNA extraction from formalin-fixed paraffin-embedded samples

INTRODUÇÃO: O desenvolvimento de métodos de extração de RNA a partir de amostras fixadas em formalina e embebidas em parafina (FFEP) possibilitou estudos retrospectivos de biologia molecular. Objetivos: Comparar a quantidade e a qualidade do RNA extraído de amostras FFEP a partir de três kits disponíveis comercialmente. MATERIAL E MÉTODO: Utilizando-se três diferentes procedimentos, o RNA total foi extraído de 14 blocos de parafina contendo fragmentos de carcinomas mamários, todos arquivados há 10 anos. A quantidade do RNA foi expressa em pg/µl; e a qualidade, pelo número de integridade do RNA (NIR), utilizando-se o Bioanalyzer da Agilent com o Pico LabChip. O RNA de maior NIR extraído de cada uma das 14 amostras foi amplificado por reação em cadeia da polimerase com transcrição reversa (RT-PCR) utilizando-se o gene G6PD, com primers designados para gerar fragmentos de 67, 151 e 242 pares de bases (pb). RESULTADOS: A média e a mediana da quantidade do RNA extraído para os três protocolos foram, respectivamente, 42,91 e 31,31 pg/µl. A média e a mediana do NIR foram, respectivamente, 1,8 e 2. Em todas as amostras, o gene G6PD foi amplificado para fragmentos de RNA de 67 e 151 pb. DISCUSSÃO: Como houve grande variação individual na quantidade e na qualidade do RNA extraído para cada amostra, os dados do presente estudo indicam que, se não for possível extrair RNA de uma determinada amostra na primeira tentativa, uma segunda extração deve ser realizada antes de se descartar essa amostra para testes de biologia molecular. CONCLUSÕES: Nos três procedimentos utilizados foi possível extrair RNA de qualidade aceitável para amplificação por RT-PCR (com NIR > 1,4).BACKGROUND: The development of methods for RNA extraction from formalin-fixed paraffin-embedded (FFPE) samples has allowed retrospective studies of molecular biology. OBJECTIVES: To compare the quantity and quality of RNA extracted from FFPE samples using three commercially available kits. Material and methods: Using three different procedures, the total RNA was extracted from 14 paraffin blocks containing fragments of mammary carcinomas, which had been archived for 10 years. The quantity of RNA was expressed in pg/µl; and the quality in RNA integrity number (RIN), by using the Agilent Bioanalyser with Pico LabChip. The RNA with higher RIN extracted from each of the 14 samples was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using the G6PD gene with primers designed to create fragments with 67, 151 and 242 base pairs (bp). RESULTS: The mean and median of RNA quantity extracted for the three procedures were respectively 42.91 and 31.31 pg/µl. The mean and median of RIN were respectively 1.8 and 2. In all the samples, the G6PD gene was amplified for RNA fragments with 67 and 151 bp. DISCUSSION: Due to the significant individual variation in quantity and quality of the extracted RNA from each sample, the data from the present study show that, if it is not possible to extract RNA from a given sample in the first attempt, a second extraction should be performed before excluding this sample. CONCLUSION: It was possible to extract RNA with acceptable quality for amplification by RT-PCR (RIN > 1.4) in the three procedures used.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

EVALUATION OF PRESENTATION OF SIGNS AND SYMPTOMS OF FEMOROACETABULAR IMPINGEMENT AFTER EPIPHYSIOLYSIS OF THE PROXIMAL FEMUR

ABSTRACTThe long-bone fractures occur most frequently in the tibial shaft. Adequate treatment of such fractures avoids consolidation failure, skewed consolidation and reoperation. To classify these fractures, the AO/OTA classification method is still used, but it is worthwhile getting to know the Ellis classification method, which also includes assessment of soft-tissue injuries. There is often an association with compartmental syndrome, and early diagnosis can be achieved through evaluating clinical parameters and constant clinical monitoring. Once the diagnosis has been made, fasciotomy should be performed. It is always difficult to assess consolidation, but the RUST method may help in this. Radiography is assessed in two projections, and points are scored for the presence of the fracture line and a visible bone callus. Today, the dogma of six hours for cleaning the exposed fracture is under discussion. It is considered that an early start to intravenous antibiotic therapy and the lesion severity are very important. The question of early or late closure of the lesion in an exposed fracture has gone through several phases: sometimes early closure has been indicated and sometimes late closure. Currently, whenever possible, early closure of the lesion is recommended, since this diminishes the risk of infection. Milling of the canal when the intramedullary nail is introduced is still a controversial subject. Despite strong personal positions in favor of milling, studies have shown that there may be some advantage in relation to closed fractures, but not in exposed fractures

Um estudo retrospectivo dos achados histopatológicos em 894 casos de megacólon: qual é a relação entre megacólon e o câncer de cólon?

Patients with megaesophagus (ME) have increased prevalence of cancer of the esophagus. In contrast, a higher incidence of colorectal cancer is not observed in patients with megacolon (MC). MC is very common in some regions of Brazil, where it is mainly associated with Chagas disease. We reviewed the pathology records of surgical specimens of all patients submitted for surgical resection of MC in the Hospital das Clínicas of the Faculty of Medicine of Ribeirão Preto (HC-FMRP), from the University of São Paulo. We found that 894 patients were operated from 1952 until 2001 for MC resection. Mucosal ulcers, hyperplasia and chronic inflammation were frequently found, while polyps were uncommon. No patients with MC presented any type of colonic neoplasm. This observation reinforces the hypothesis that MC has a negative association with cancer of the colon. This seems to contradict the traditional concept of carcinogenesis in the colon, since patients with MC presents important chronic constipation that is thought to cause an increase in risk for colon cancer. MC is also associated with other risk factors for cancer of colon, such as hyperplasia, mucosal ulcers and chronic inflammation. In ME these factors lead to a remarkable increase in cancer risk. The study of mucosal cell proliferation in MC may provide new insights and useful information about the role of constipation in colonic carcinogenesis.Pacientes com megaesôfago (ME) possuem incidência aumentada de câncer de esôfago. Em contraste, há poucos relatos na literatura de associação entre megacólon (MC) e câncer de cólon. O MC é muito comum em algumas regiões do Brasil, e na maioria das vezes, está associado à Doença de Chagas. Nós reavaliamos os arquivos de patologia de peças cirúrgicas de todos os pacientes submetidos à ressecção de MC no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HC-FMRP), da Universidade de São Paulo. Encontramos o número de 894 pacientes que foram operados de 1952 até 2001 para a ressecção do MC. Úlceras da mucosa, hiperplasia e inflamação crônica foram frequentemente encontrados, enquanto pólipos foram incomuns. Nenhum paciente com MC apresentou qualquer tipo de neoplasia do cólon. Essa observação reforça a hipótese de que o MC tem uma associação negativa com câncer de cólon. Isso parece contradizer o conceito tradicional de carcinogênese do cólon, uma vez que pacientes com MC apresentam constipação crônica importante, a qual é tida como uma causa que aumenta o risco de câncer de cólon. MC também está associado a outros fatores de risco para o câncer de cólon, como hiperplasia, úlceras da mucosa e inflamação crônica. No megaesôfago (ME), tais fatores aumentam o risco de câncer esofágico. O estudo da proliferação celular da mucosa no MC pode fornecer informações úteis sobre o papel da constipação na carcinogênese colônica

A imunomarcação positiva para c-kit está associada com a presença de células análogas às intersticiais de Cajal no músculo ciliar?

PURPOSE: Interstitial cells of Cajal were identified in the gastrointestinal tract of several species, with close relation to the enteric nervous system. Since it was recognized that interstitial cells of Cajal express the gene product of c-kit, we performed immunohistochemistry for c-kit protein in ciliary muscle specimens of monkeys' eyes. METHODS: Eight eyes from four adult male new world monkeys (Cebus apella) were studied. After blocking endogenous peroxidase activity and nonspecific protein binding, 1:100 dilution of mouse monoclonal antibody against c-kit human oncoprotein was applied to tissues. Antigen-antibody reaction was visualized using the avidin-biotinylated horseradish peroxidase complex in each slide. RESULTS: We observed some groups of fusiform c-kit expressing cells located amongst muscle bundles of the ciliary muscle. Other pigment cells and mast cells were also observed. CONCLUSION: C-kit expressing cells observed in the ciliary muscle of Cebus apella, showed no similarity to melanocytes or mast cells and they could be associated with their gastrointestinal interstitial cells of Cajal counterpart.OBJETIVO: As células intersticiais de Cajal estão presentes no trato gastrintestinal de diversas espécies animais, em íntima relação com o sistema nervoso entérico. Uma vez que as células intersticiais de Cajal expressam o produto do gene c-kit, realizou-se um ensaio imuno-histoquímico a fim de se verificar a marcação da proteína c-kit no músculo ciliar de amostras de olhos de macacos. MÉTODOS: Oito olhos de quatro macacos do novo mundo (Cebus apella) foram estudados. Após bloqueio da peroxidase endógena e de ligação protéica não específica, os tecidos receberam aplicação de anticorpos de camundongos antioncoproteína c-kit humana (1:100). A reação antígeno-anticorpo foi verificada através da aplicação do complexo avidina-biotinilada-peroxidase em cada lâmina. RESULTADOS: Foram observados grupos de células que expressam c-kit, localizadas entre as fibras do músculo ciliar. Mastócitos e outras células pigmentadas também foram observadas. CONCLUSÃO: Algumas células que expressam c-kit, observadas no músculo ciliar de Cebus apella, não mostraram similaridade com mastócitos ou melanócitos e podem ser classificadas como análogas das células intersticiais de Cajal gastrintestinais.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)FAPES

INITIAL GROWTH OF Citharexyllum myrianthum plants UNDER PLANTING FERTILIZATION

The objective of this work was to evaluate the initial growth of Citharexyllum myrianthum under the influence of three sources of planting fertilizers, in conditions of 18 L pots. The treatments used were, as follows: absolute control; 5 L of biosolid/pot; 300 g of natural rock phosphate (NRP), and 200 grams of NPK 06-29-06. The seedlings were grown in pots in full sun, supplemented with Dystrophic Red Yellow Latosol. At 150 days after transplanting, the plants were measured, obtaining the height of the aerial part and the diameter of the stem. Then, the increment of these two variables was calculated. Subsequently, the leaves were removed to determine the leaf area (LA), separating the aerial part from the root system, and then packed in a paper bag and placed in an oven at 65ºC, for drying until reaching constant weight. After weighing this material to obtain the dry matter of the aerial part (APDM) and root (RDM). The leaves were ground to determine the foliar contents of N, P, K, Ca, and Mg. To evaluate the effect of treatments on the soil, samples were taken from the pots to determine the levels of macronutrients, organic matter, and CEC(t). It was found that the LA and APDM of plants fertilized with NPK showed significantly higher values when compared to plants cultivated in the control treatment. It is recommended to fertilize Citharexyllum myrianthum plants with 200 grams of N-P-K 06-29-06/hole or with 5 L of biosolid

Tendência e Características da Sífilis Congênita e Materna no Rio de Janeiro: 2007-2017: Trend and Characteristics of Congenital and Maternal Syphilis in Rio de Janeiro: 2007-2017

Introduction: Our aim was to describe the profile of congenital (CS), gestational syphilis (GS) notifications and temporal analysis of the incidence rate in the SINAN database, between 2007 and 2017 in Rio de Janeiro State. Methods: This quantitative, descriptive study used data from CS-SINAN and GS-SINAN databases. The temporal analysis was done through polynomial regression models using incidence rate annual distribution. Results: In CS-SINAN we observed progressive notification; 93.4% were from children up to six days old and lethality rate was 2.4%. It was found that 46,5% of CS cases’ mothers detected syphilis at prenatal and only 11.8% of sexual partners were treated. In GS-SINAN there was growth in notification and predominated women aged 20 to 30 years old, earlier diagnosed in prenatal care over the years. We found reduction of vertical transmission through the years. The temporal analysis of the incidence rates revealed a trend of growth in both CS and GS. Conclusion: Our data suggests that the increased detection in GS has slowed the spread of CS, yet the progressive annual increase in case number demonstrates that syphilis is not controlled, therefore, measures to ensure prenatal care, sexual partner treatment and health care training should be prioritized.Objetivo: Describir el perfil de las notificaciones de sífilis congénita (CS) y sífilis gestacional (SG) y el análisis temporal de las tasas de incidencia en la base de datos del SINAN, de 2007 a 2017 en el Estado de Río de Janeiro. Métodos: Estudio cuantitativo, descriptivo, se utilizó información del SC-SINAN y SG-SINAN. El análisis temporal probó modelos de regresión polinómica utilizando la distribución de la tasa de incidencia anual. Resultados: En SC-SINAN hay un aumento progresivo de notificaciones; El 93,4% eran niños ≤6 días y la tasa de letalidad fue del 2,4%. El 46,5% de las madres de casos de SC diagnosticó sífilis en el control prenatal y solo el 11,8% de las parejas la trató. En el SG-SINAN crecieron las notificaciones, principalmente en mujeres de 20 a 30 años, con un diagnóstico cada vez más precoz a lo largo de los años. Hubo una reducción en la transmisión vertical. El análisis temporal de la tasa de incidencia revela una tendencia al alza tanto para SC cuanto para SG. Conclusión: estos datos sugieren que el aumento en la detección de GS redujo la expansión de SC, aunque el aumento anual en el número de casos demuestra que la sífilis no se controla, por lo tanto, medidas que garanticen la atención prenatal, el tratamiento de la pareja sexual y la capacitación en salud. debe ser priorizado.Objetivo: Descrever o perfil das notificações de sífilis congênita (SC) e sífilis gestacional (SG) e da análise temporal das taxas de incidência no banco de dados do SINAN, no período de 2007 a 2017 no Estado do Rio de Janeiro. Métodos: Estudo quantitativo, descritivo utilizou informações do SC-SINAN e SG-SINAN. A análise temporal testou modelos de regressão polinomial utilizando a distribuição anual da taxa de incidência. Resultados: No SC-SINAN observa-se aumento progressivo nas notificações; 93,4% eram crianças ≤ 6 dias e taxa de letalidade foi 2,4%. Das mães dos casos de SC, 46,5% foram diagnosticadas com sífilis no pré-natal e apenas 11,8% dos parceiros trataram. No SG-SINAN, as notificações cresceram, principalmente em mulheres de 20 a 30 anos, com o diagnóstico cada vez mais precoce com o passar dos anos. Houve redução na transmissão vertical. A análise temporal da taxa de incidência revela uma tendência de crescimento tanto da SC quanto SG. Conclusão: Estes dados sugerem que o aumento na detecção da SG reduziu a expansão da SC, ainda que o aumento anual no número de casos demonstre que a sífilis não está controlada, portanto medidas que garantam a assistência pré-natal, o tratamento do parceiro sexual e o treinamento em saúde deveriam ser priorizados

Triple-blinded randomized clinical trial comparing efficacy and tooth sensitivity of in-office and at-home bleaching techniques

Objective: Our study aims to compare the efficacy and tooth sensitivity following in-office (35% hydrogen peroxide) or at-home (10% carbamide peroxide) bleaching treatments both preceded by 2% potassium nitrate (2%KF) desensitizing gel. Methodology: 130 volunteers were randomly allocated to a) in-office bleaching and a placebo at-home protocol; or b) in-office placebo and at-home bleaching treatment. 2% KF was applied for 10 min before both treatments. Objective: color evaluation was performed (spectrophotometer CIEL*a*b* system and CIEDE2000) to calculate the color change (ΔE00). Subjective evaluation was performed using the VITA classical shade guide followed by shade variation (ΔSGU) at the beginning and end of bleaching treatment and 2 weeks post-bleaching. Tooth sensitivity was daily recorded using a Likert scale varying from 1 (no sensitivity) to 5 (severe sensitivity). Analysis was carried out using non-parametric tests. Results: Regarding the color change, at-home bleaching resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.003) and Δa* (p=0.014). Two weeks post-bleaching, the at-home treatment resulted in significant color improvement compared to in-office treatment for the parameters Δb* (p=0.037) and ΔE00 (p=0.033). No differences were observed in either ΔSGU parameters. Concerning sensitivity, patients treated with in-office bleaching reported more tooth sensitivity than the at-home group only on the first day after bleaching started, without significant differences in the other periods evaluated (p>0.05). Conclusions: At-home and in-office bleaching, preceded by a desensitizing agent, were effective for vital teeth bleaching and 10% carbamide peroxide produced a higher whitening effect than 35% hydrogen peroxide in the short time evaluation. Tooth sensitivity rates were similar for the two techniques tested

Evaluation of terminal vertebral plate on cervical spine at different age groups and its correlation with intervertebral disc thickness

OBJETIVO: Avaliar por meio de histomorfometria, a espessura da placa vertebral terminal, a espessura do disco intervertebral e a sua correlação nas diferentes faixas etárias, tentando identificar a sua correlação. MÉTODOS: Foram avaliados os segmentos cervicais C4-C5 e C5-C6 de 50 cadáveres humanos , de ambos os sexos, divididos em 5 grupos com intervalo de faixa etária de 10 anos, a partir dos 21 anos de idade. A avaliação da espessura da PVT e do disco intervertebral foi realizada por meio de avaliação histomorfométrica dos cortes histológicos corados pela hematoxilina e eosina. A espessura da PVT inferior de C4, PVT superior de C5, PVT inferior de C5 e PVT superior de C6 foram comparadas entre si e também com a espessura do disco intervertebral interposto entre as respectivas PVT. RESULTADOS: A espessura das placas vertebrais terminais adjacentes ao mesmo DI não apresentou diferença estatística. No entanto, a comparação da espessura das placas vertebrais superior e inferior da mesma vértebra cervical (C5), apresentou diferença estatística em todas as faixas etárias avaliadas. Foi observado coeficiente de correlação estatística, maior que 80%, entre a espessura daplaca vertebral terminal e o disco intervertebral adjacente, com proporcional redução de espessura de ambas estruturas, nos diferentes níveis cervicais avaliados, e também nas diferentes faixas etárias avaliadas. CONCLUSÃO: A placa vertebral terminal apresenta correlação morfológica com o disco intervertebral ao qual entra em contato, e não apresenta correlação com a placa vertebral terminal da mesma vértebra.OBJECTIVE: To evaluate, by means of histomorphometry, terminal vertebral plate thickness, intervertebral disc thickness and its correlation on different age groups, seeking to identify its correlation. METHODS: C4-C5 and C5-C6 cervical segments removed from human cadavers of both genders were assessed and divided into five groups of 10-year age intervals, from 21 years old. TVP and intervertebral disc thickness evaluation was made by means of histomorphometry of histological slides stained with hematoxylin and eosyn. Lower C4 TVP, upper C5 TVP, and upper C6 TVP de were compared between each other and to the interposed intervertebral disc thickness between relevant TVP. RESULTS: The thickness of terminal vertebral plates adjacent to the same ID did not show statistic differences. However, the comparison of upper and lower vertebral plates thickness on the same cervical vertebra (C5), showed statistical difference on all age groups studied. We found a statistical correlation coefficient above 80% between terminal vertebral plate and adjacent intervertebral disc, witha proportional thickness reduction of both structures on the different cervical levels studied, and also on the different age groups assessed. CONCLUSION: Terminal vertebral plate shows a morphologic correlation with the intervertebral disc next to it, and does not show correlation with the terminal vertebral plate on the same vertebra

Articulando raça, gênero e sexualidade para pensar a construção social das múltiplas masculinidades negras

Apresentação do dossiê temático Masculinidades Negras

Guidelines of the Brazilian Association of Studies on Alcohol and Other Drugs (ABEAD) for diagnoses and treatment of psychiatric comorbidity with alcohol and other drugs dependence

Recently, several studies have focused on comorbity psychiatric disorders with alcohol and other substance dependence. The Brazilian Association of Studies on Alcohol and Other Drugs proposed the Brazilian Guidelines project. This study review diagnostic and therapeutic criteria to the most prevalent psychiatric comorbidities. Randomized clinical trials, epidemiological, animal studies and other forms of research are reviewed. The main psychiatric comorbidities are studied based on guidelines adopted by other countries and the literature data resumed. Epidemiological aspects, diagnoses, integrated treatment and service organization, as well as specific psychotherapic and pharmacological treatment are discussed. The Brazilian Association of Studies on Alcohol and Other Drugs Guidelines reassures the importance of adequate diagnoses and treatment regarding alcoholic and drug dependent patients suffering of comorbid psychiatric disorders.O diagnóstico e tratamento de comorbidade psiquiátrica e dependência de álcool e outras substâncias tem sido objeto de inúmeros estudos nos últimos anos. A Associação Brasileira de Estudos do Álcool e Outras Drogas desenvolveu o projeto Diretrizes. Este trabalho visa o desenvolvimento de critérios diagnósticos e terapêuticos atualizados para as comorbidades psiquiátricas mais prevalentes. Ensaios clínicos randomizados, estudos epidemiológicos, com animais e outros estudos são revisados. As principais comorbidades psiquiátricas são estudadas e os dados de literatura resumidos, tendo como referência diretrizes adotadas em outros países. São abordados aspectos epidemiológicos, critérios diagnósticos, tratamento integrado e organização de serviço especializado, assim como especificidades do tratamento psicoterápico e farmacológico. As Diretrizes da Associação Brasileira de Estudos do Álcool e Outras Drogas reforçam a importância da abordagem adequada do dependente químico portador de comorbidade psiquiátrica.Universidade Federal de Santa Catarina Núcleo de PsiquiatriaInstituto de Psiquiatria de Santa CatarinaUniversidade Federal de São Paulo (UNIFESP) Unidade de Pesquisa em Álcool e DrogasSanta Casa de Misericórdia de São Paulo Unidade de Álcool e DrogasUniversidade de São Paulo Faculdade de Medicina Hospital das ClinicasCentro de Atendimento Médico e SocialHospital de Clínicas de Porto AlegreHospital Israelita Albert EinsteinSanta Casa do Rio de Janeiro Setor de Dependência QuímicaUniversidade Gama FilhoUNIFESP, Unidade de Pesquisa em Álcool e DrogasSciEL