Autism in Brazil: a systematic review of family challenges and coping strategies / Autismo no Brasil, desafios familiares e estratégias de superação: revisão sistemática

OBJECTIVE: To describe the challenges faced by families caring for children with autism spectrum disorder (ASD) in Brazil and the coping strategies employed. SOURCES: Systematic review of articles published until September of 2013, without language restrictions, using quality appraisal (AMSTAR and CASP/Oxford instruments). SUMMARY OF THE FINDINGS: The literature shows parental emotional overload as one of the main challenges faced by families, especially mothers. The main stressors were diagnostic postponement, difficulty dealing with the diagnosis and associated symptoms, and poor access to health services and social support. The predominant coping strategies found included information exchange between affected families and integrated healthcare network for patient and family support. CONCLUSION: ASD exerts strong influence on family dynamics, resulting in caregiver overload, especially in mothers. The Brazilian Unified Health System needs to provide comprehensive, continuous, and coordinated care to strengthen the patient-family dyad and promote the full development and societal inclusion of children with ASD. .

OBJETIVO: Descrever os desafios encontrados pelas famílias na convivência com crianças portadoras de transtorno do espectro autista (TEA) no Brasil e as estratégias de superação empregadas. FONTE DOS DADOS: Revisão sistemática da literatura com inclusão de artigos publicados até setembro de 2013, sem restrições de idioma. Os artigos incluídos foram submetidos à avaliação de qualidade metodológica por meio do Amstar e Casp/Oxford. SÍNTESE DOS DADOS: Incluem-se estudos provenientes de São Paulo e Rio Grande do Sul com alta e moderada qualidade metodológica. A literatura mostra sobrecarga emocional dos pais como um dos principais desafios encontrados pelas famílias, inclusive com grande tensão sobre as mães. Dentre os fatores relacionados ao estresse estão: postergação diagnóstica, dificuldade de lidar com o diagnóstico e com os sintomas associados, acesso precário ao serviço de saúde e apoio social. Dentre as estratégias de superação destacaram-se: troca de informações entre as famílias afetadas e assistência integralizada da rede de saúde no atendimento do paciente e suporte à família. CONCLUSÃO: Observou-se que o TEA exerce forte influência na dinâmica familiar com sobrecarga dos cuidadores, geralmente da mãe. O Sistema Único de Saúde necessita prover cuidado integral, longitudinal e coordenado com vistas ao fortalecimento do binômio paciente-família e o pleno desenvolvimento e a plena inserção dessas crianças na sociedade. .

Dose-Response and Time-Course of a-Tocoferol Mediating the Cytoprotection Of Dental Pulp Cells Against Hydrogen Peroxide

This in vitro study evaluated the potential protective effect of vitamin E alpha-tocopherol (α-T) isomer against the toxicity of hydrogen peroxide (HP) applied on dental pulp cells. Odontoblast-like MDPC-23 cells were seeded on 96-well plates for 72 h, treated with different concentrations of α-T (1, 3, 5, and 10 mM) for different times (1, 4, 8, and 24 h) and then exposed or not to a 0.018% HP solution for 30 min. In positive and negative control groups, cells were exposed to HP or culture medium (DMEM containing 5% DMSO), respectively. Cell viability was assessed by the MTT assay and the absorbance numeric data, expressed as percentage values, were subjected to the statistical analysis by Kruskal-Wallis and Mann-Whitney tests (α=5%). Considering the cells in the negative control as having 100% of cell viability, all combinations of α-T concentrations and pretreatment times showed a protective effect against HP cytotoxicity. Significant reduction of cell viability (59%) was observed in the positive control compared with the negative control. The highest values of pulp cell viability were obtained after pretreatment with 1 and 3 mM α-T concentrations for 24 h followed by exposure to HP (126% and 97% of cell viability, respectively). Under the tested conditions, the most effective cell protection against the cytotoxic effects of HP was provided by the lowest concentrations of α-T (1 and 3 mM) applied for 24 h.

Neste estudo, foi avaliado o potencial protetor do isômero alfa-tocoferol da vitamina E (-T) contra a ação tóxica do peróxido de hidrogênio (PH) aplicado sobre células pulpares. Células odontoblastóides MDPC-23 foram semeadas em placas de 96 compartimentos por 72 h, tratadas com diferentes concentrações de α-T (1, 3, 5 e 10 mM) por diferentes períodos (1, 4, 8 e 24 h) e, então, expostas ou não a uma solução com 0,0018% de PH por 30 min. Nos controles positivo e negativo, as células foram expostas ao PH ou meio de cultura (DMEM contendo 5% de DMSO), respectivamente. A viabilidade celular foi avaliada pelo teste do MTT e os valores numéricos de absorbância, expressos em porcentagem, foram submetidos a análise estatística pelos testes de Kruskal-Wallis e Mann-Whitney (α=5%). Considerando as células do grupo controle negativo como apresentando 100% de viabilidade celular, todas as combinações de α-T, nas diferentes concentrações, e tempos de pré-tratamento demonstraram um efeito protetor contra a citotoxicidade do PH. Redução significativa da viabilidade (59%) foi observada para o grupo controle positivo comparado ao controle negativo. Os maiores valores de viabilidade celular foram obtidos após pré-tratamento com 1 e 3 mM de α-T por 24 h seguido de exposição ao PH (126% e 97% de viabilidade celular, respectivamente). Assim, de acordo com as condições experimentais, o efeito protetor mais efetivo contra os efeitos tóxicos do PH foi observado para as menores concentrações de α-T (1 e 3 mM) aplicado por 24 h.

Sexual hormones modulate compensatory renal growth and function / Las hormonas sexuales modulan el crecimiento renal compensador (CRC) y la función renal en la uninefrectomía (uNx)

The role played by sexual hormones and vasoactive substances in the compensatory renal growth (CRG) that follows uninephrectomy (uNx) is still controversial. Intact and gonadectomized adult Wistar rats of both sexes, with and without uNx, performed at 90 days age, were studied at age 150 days. Daily urine volume, electrolyte excretion and kallikrein activity (UKa) were determined. Afterwards, glomerular filtration rate and blood pressure were measured, the kidneys weighed and DNA, protein and RNA studied to determine nuclei content and cell size. When the remnant kidney weight at age 150 days was compared with the weight of the kidney removed at the time of uNx, male uNx rats showed the greatest CRG (50%) while growth in the other uNx groups was 25%, 15% and 19% in orchidectomized, female and ovariectomized rats, respectively. The small CRG observed in the uNx female rats was accompanied by the lowest glomerular filtration value, 0.56 ± 0.02 ml/min/g kwt compared, with the other uNx groups, p < 0.05. Cell size (protein or RNA/DNA) was similar for all the groups except for uNx orchidectomized rats. In this group the cytoplasmatic protein or RNA content was lower than in the other groups while DNA (nuclei content) was similar. Some degree of hyperplasia was determined by DNA content in the uNx groups. Male sexual hormones positively influenced CRG and its absence modulated cell size. Female sexual hormones, instead, did not appear to stimulate CRG. The kallikrein kinin system may not be involved in CRG.

La importancia que pueden tener las hormonas sexuales y sustancias vasoactivas sobre el crecimiento renal compensador (CRC) que sigue a la uninefrectomía es aún materia de debate. Se estudiaron ratas Wistar de ambos sexos, a los 150 días de vida, intactas y gonadectomizadas con y sin uNx, realizada a los 90 días de vida. Se midió volumen urinario diario y excreción de electrolitos y actividad de kalikreína urinaria. Se midió filtrado glomerular y presión arterial media extrayéndose luego los riñones que fueron pesados y preparados para estudios histológicos y determinación de ADN, ARN y proteínas para estimar contenido nuclear y tamaño celular. El CRC fue calculado comparando el peso del riñón al momento de las uNx (90 dias de vida) con aquel obtenido a los 150 días de vida. En las ratas macho uNx se observó el mayor CRC (50%) mientras que, en los otros grupos uNx solo alcanzó un 25%, 15% y 19%. El filtrado glomerular acompañó los cambios morfológicos observándose el menor filtrado en las ratas hembras uNx respecto al resto de los grupos 0.56 ± 0.02, p < 0.05. El tamaño celular (proteína o ARN/ ADN) fue similar para todos los grupos excepto para los orquidectomizados uNx, cuyo contenido citoplasmático fue menor. El contenido nuclear (ADN) fue semejante en todos los grupos. Se observó que el CRC está influenciado positivamente por las hormonas sexuales masculinas y su ausencia modula el tamaño celular. La falta de hormonas sexuales femeninas, en cambio, afecta negativamente el CRC. El sistema kalikreína kinina no parecería estar involucrado en el CRC.

Relationships between dendritic morphology, spatial distribution and firing patterns in rat layer 1 neurons

The cortical layer 1 contains mainly small interneurons, which have traditionally been classified according to their axonal morphology. The dendritic morphology of these cells, however, has received little attention and remains ill defined. Very little is known about how the dendritic morphology and spatial distribution of these cells may relate to functional neuronal properties. We used biocytin labeling and whole cell patch clamp recordings, associated with digital reconstruction and quantitative morphological analysis, to assess correlations between dendritic morphology, spatial distribution and membrane properties of rat layer 1 neurons. A total of 106 cells were recorded, labeled and subjected to morphological analysis. Based on the quantitative patterns of their dendritic arbor, cells were divided into four major morphotypes: horizontal, radial, ascendant, and descendant cells. Descendant cells exhibited a highly distinct spatial distribution in relation to other morphotypes, suggesting that they may have a distinct function in these cortical circuits. A significant difference was also found in the distribution of firing patterns between each morphotype and between the neuronal populations of each sublayer. Passive membrane properties were, however, statistically homogeneous among all subgroups. We speculate that the differences observed in active membrane properties might be related to differences in the synaptic input of specific types of afferent fibers and to differences in the computational roles of each morphotype in layer 1 circuits. Our findings provide new insights into dendritic morphology and neuronal spatial distribution in layer 1 circuits, indicating that variations in these properties may be correlated with distinct physiological functions.

Morfologia e contratilidade em cardiomiócitos de ratos com baixo desempenho para o exercício físico / Morphology and contractility in cardiomyocytes of rats with low exercise performance

FUNDAMENTO: A capacidade aeróbica é fundamental para o desempenho físico, e a baixa capacidade aeróbica está relacionada ao desencadeamento de diversas doenças cardiovasculares. OBJETIVO: Comparar a contratilidade e a morfologia de cardiomiócitos isolados de ratos com baixo desempenho e desempenho padrão para o exercício físico. MÉTODOS: Ratos Wistar, com 10 semanas de idade, foram submetidos a um protocolo de corrida em esteira até a fadiga, e foram divididos em dois grupos: Baixo Desempenho (BD) e Desempenho Padrão (DP). Em seguida, após eutanásia, o coração foi removido rapidamente e, por meio de dissociação enzimática, os cardiomiócitos do ventrículo esquerdo foram isolados. O comprimento celular e dos sarcômeros e a largura dos cardiomiócitos foram medidos usando-se um sistema de detecção de bordas. Os cardiomiócitos isolados foram estimulados eletricamente a 1 e 3 Hz e a contração celular foi medida registrando-se a alteração do seu comprimento. RESULTADOS: O comprimento celular foi menor no grupo BD (157,2 ± 1,3µm; p < 0,05) em relação ao DP (161,4 ± 1,3 µm), sendo o mesmo resultado observado para o volume dos cardiomiócitos (BD, 25,5 ± 0,4 vs. DP, 26,8 ± 0,4 pL; p < 0,05). Os tempos para o pico de contração (BD, 116 ± 1 vs. DP, 111 ± 2ms) e para o relaxamento total (BD, 143 ± 3 vs. DP, 232 ± 3 ms) foram maiores no grupo BD. CONCLUSÃO: Conclui-se que os miócitos do ventrículo esquerdo dos animais de baixo desempenho para o exercício físico apresentam menores dimensões que os dos animais de desempenho padrão, além de apresentarem perdas na capacidade contrátil.

BACKGROUND: Aerobic capacity is essential to physical performance, and low aerobic capacity is related to the triggering of various cardiovascular diseases. OBJECTIVE: To compare the morphology and contractility of isolated rat cardiomyocytes with low performance and standard performance for exercise. METHODS: Wistar rats with 10 weeks of age underwent a protocol of treadmill running to fatigue, and were divided into two groups: Low Performance (LP) and Standard Performance (SP). Then, the animals were sacrificed, the heart was quickly removed and, by means of enzymatic dissociation, left ventricular cardiomyocytes were isolated. The cell and sarcomeres length and width of cardiomyocytes were measured using an edge detection system. The isolated cardiomyocytes were electrically stimulated at 1 and 3 Hz and cell contraction was measured by registering the change of their length. RESULTS: The cell length was shorter in the LP group (157.2 1.3µm; p < 0.05) compared to SP (161.4 1.3µm), and the same result was observed for the volume of cardiomyocytes (LP, 25.5 0.4. vs. SP, 26.8 ± 0.4 pL; p < 0.05). The time to peak contraction (LP, 116 1 vs. SP 111 2ms) and total relaxation (LP, 143 3 vs. SP 232 3ms) were higher in the LP group. CONCLUSION: We conclude that left ventricular myocytes of animals with low performance for exercise are smaller than animals with standard performance. In addition to that, they present losses in contractile capacity.

Optimization and comparison of two different 3D culture methods to prepare cell aggregates as a bioink for organ printing

The ultimate goal of tissue engineering is to design and fabricate functional human tissues that are similar to natural cells and are capable of regeneration. Preparation of cell aggregates is one of the important steps in 3D tissue engineering technology, particularly in organ printing. Two simple methods, hanging drop (HD) and conical tube (CT) were utilized to prepare cell aggregates. The size and viability of the aggregates obtained at different initial cell densities and pre-culture duration were compared. The proliferative ability of the cell aggregates and their ability to spread in culture plates were also investigated. In both methods, the optimum average size of the aggregates was less than 500 µm. CT aggregates were smaller than HD aggregates. 5,000 cells per drop HD aggregates showed a marked ability to attach and spread on the culture surface. The proliferative ability reduced when the initial cell density was increased. Comparing these methods, we found that the HD method having better size controlling ability as well as enhanced ability to maintain higher rates of viability, spreading, and proliferation. In conclusion, smaller HD aggregates might be a suitable choice as building blocks for making bioink particles in bioprinting technique.

Measuring bacterial cells size with AFM

Atomic Force Microscopy (AFM) can be used to obtain high-resolution topographical images of bacteria revealing surface details and cell integrity. During scanning however, the interactions between the AFM probe and the membrane results in distortion of the images. Such distortions or artifacts are the result of geometrical effects related to bacterial cell height, specimen curvature and the AFM probe geometry. The most common artifact in imaging is surface broadening, what can lead to errors in bacterial sizing. Several methods of correction have been proposed to compensate for these artifacts and in this study we describe a simple geometric model for the interaction between the tip (a pyramidal shaped AFM probe) and the bacterium (Escherichia coli JM-109 strain) to minimize the enlarging effect. Approaches to bacteria immobilization and examples of AFM images analysis are also described.

Cardiac and renal effects induced by different exercise workloads in renovascular hypertensive rats

We examined the effect of exercise training (Ex) without (Ex 0 percent) or with a 3 percent workload (Ex 3 percent) on different cardiac and renal parameters in renovascular hypertensive (2K1C) male Fisher rats weighing 150-200 g. Ex was performed for 5 weeks, 1 h/day, 5 days/week. Ex 0 percent or Ex 3 percent induced similar attenuation of baseline mean arterial pressure (MAP, 119 ± 5 mmHg in 2K1C Ex 0 percent, N = 6, and 118 ± 5 mmHg in 2K1C Ex 3 percent, N = 11, vs 99 ± 4 mmHg in sham sedentary (Sham Sed) controls, N = 10) and heart rate (HR, bpm) (383 ± 13 in 2K1C Ex 0 percent, N = 6, and 390 ± 14 in 2K1C Ex 3 percent, N = 11 vs 371 ± 11 in Sham Sed, N = 10,). Ex 0 percent, but not Ex 3 percent, improved baroreflex bradycardia (0.26 ± 0.06 ms/mmHg, N = 6, vs 0.09 ± 0.03 ms/mmHg in 2K1C Sed, N = 11). Morphometric evaluation suggested concentric left ventricle hypertrophy in sedentary 2K1C rats. Ex 0 percent prevented concentric cardiac hypertrophy, increased cardiomyocyte diameter and decreased cardiac vasculature thickness in 2K1C rats. In contrast, in 2K1C, Ex 3 percent reduced the concentric remodeling and prevented the increase in cardiac vasculature wall thickness, decreased the cardiomyocyte diameter and increased collagen deposition. Renal morphometric analysis showed that Ex 3 percent induced an increase in vasculature wall thickness and collagen deposition in the left kidney of 2K1C rats. These data suggest that Ex 0 percent has more beneficial effects than Ex 3 percent in renovascular hypertensive rats.

Influência do substrato e do tamanho da célula de bandejas de poliestireno expandido no desenvolvimento de mudas e produção de calêndula (Calendula officinalis L. ) / Influence of substrate and cell size of expanded polystyrene tray on the development and production of marigold (Calendula officinalis L. ) seedlings

A calêndula (Calendula officinalis L.) é uma planta medicinal anual de origem Mediterrânica que apresenta propriedades antiinflamatórias, antivirais, antigenotóxicas. A propagação desta planta normalmente é feita por sementes, sendo o uso de bandejas de poliestireno uma forma econômica de produção de mudas. Foram conduzidos dois experimentos com o objetivo de avaliar a interferência de dois tamanhos de célula em bandeja de poliestireno expandido (40 cm³ e 12 cm³) e de dois substratos (solo + esterco bovino curtido e substrato comercial tipo Plantmax para hortaliças®) sobre a qualidade de mudas e a produção de calêndula. O delineamento estatístico nos dois experimentos foi em blocos casualizados, dispostos em arranjo fatorial 2 x 2 (dois substratos e dois tamanhos de células) com seis repetições. Os tratamentos, referentes aos tamanhos das células foram bandeja com 128 células (40 cm³ de volume) e, bandeja com 288 células (12 cm³ de volume). Os substratos utilizados foram solo de cerrado + esterco bovino curtido (1:1) e substrato comercial tipo "Plantmax para hortaliças®". Parte das mudas foi transplantada para avaliação da produção de capítulos. Houve diferenças significativas entre os substratos para as variáveis: matéria seca de parte aérea, matéria seca da raiz e matéria seca total. O uso do substrato formulado com solo de cerrado e esterco bovino proporcionou o melhor desenvolvimento das mudas de calêndula. Contudo, os diferentes tipos de substrato e bandejas de poliestireno não influenciaram significativamente a produção de capítulos das mudas transplantadas.

Marigold Calendula (Calendula officinalis L.) is an annual medicinal plant of Mediterranean origin which presents anti-inflammatory, antiviral and antigenotoxic properties. This species normally propagates through seeds, and the use of polystyrene trays is an economic way to produce its seedlings. Two experiments were carried out to evaluate the influence of two cell sizes of expanded polystyrene trays (40 cm³ and 12 cm³) and two substrates (soil + cattle manure and Plantmax® commercial substrate for vegetables) on marigold production and seedling quality. Experimental design in both assays was in randomized blocks, in a 2x2 factorial arrangement (two substrates and two cell sizes), with six replicates. Treatments regarding cell sizes were trays presenting 128 cells (40 cm³) and trays containing 288 cells (12 cm³). The used substrates were cerrado soil + cattle manure (1:1) and Plantmax® commercial substrate for vegetables. Part of the seedlings was transplanted to evaluate capitulum production. There were significant differences among substrates for total, root and shoot dry matter. The use of substrate containing cerrado soil and cattle manure led to the best development of marigold seedlings. However, the different types of substrates and polystyrene trays did not significantly influence capitulum production in transplanted seedlings.

GIST: avaliações morfológica e imuno-histoquímica do prognóstico / GIST: morphological and immunohistochemical prognostic evaluation

INTRODUÇÃO: os tumores estromais gastrointestinais possuem amplo espectro biológico, variando desde lesões de comportamento benigno até aquelas de caráter maligno, capazes de ampla disseminação e frequentes metástases viscerais. Atualmente, o prognóstico está baseado num escore, denominado grau de risco. Entretanto, este sistema apresenta falhas, com tumores classificados como de riscos intermediário e baixo associados ao desenvolvimento de metástases. Desta forma, são necessários estudos que visem ao aprimoramento desse sistema de classificação, destacando-se nesse campo, nos últimos anos, o índice de proliferação celular que tem mostrado valor prognóstico na predição da agressividade tumoral. OBJETIVOS: analisar critérios morfológicos (tamanho macroscópico, topografia do tumor, índice mitótico, necrose, subtipo histológico), verificar o grau de risco e pesquisar a aplicabilidade dos marcadores imuno-histoquímicos (actina músculo-específico, proteína S-100, Ki67 e p16ink4a) como fatores prognósticos do tumor estromal gastrointestinal (GIST). RESULTADOS: a análise univariada mostrou significância com tumores maiores que 5 cm, número de mitoses maior que 5/50 CGA, presença de necrose, de grau de risco alto e índice de proliferação celular (Ki67) maior que 5 por cento com relação à redução da sobrevida global dos pacientes (p = 0,017; 0,01; 0,001; 0,016; < 0,001 respectivamente). Os outros fatores analisados (subtipo histológico, imunofenótipo e p16ink4a) não mostraram significância. CONCLUSÃO: o grau de risco, o tamanho tumoral, o índice mitótico e a presença de necrose corroboram evidências prévias da sua utilização, como fatores morfológicos prognósticos e o emprego do índice de proliferação celular (Ki67) associado ao grau de risco para melhor esclarecimento do comportamento biológico dos GIST.

INTRODUCTION: Gastrointestinal stromal tumors (GIST) have a wide biological spectrum, ranging from benign to malignant lesions, which are prone to wide spread and frequent visceral metastasis. Currently, the prognosis is based on a score system known as risk level. However, this system has some drawbacks. For instance, tumors classified as low or intermediate risk may be associated with the development of metastasis. Therefore, studies are required to improve this classification system and incorporate recent developments such as cellular proliferation index, which has shown prognostic value in the prediction of tumor aggressiveness. OBJECTIVES: To analyze morphological criteria (macroscopic size, tumor topography, mitotic index, necrosis, histological subtype), observe risk and investigate the usefulness of immunohistochemical markers (muscle-specific actin, S-100 protein, Ki67 and p16ink4a) as prognostic markers of GIST. RESULTS: Univariate analysis showed that a reduced global survival was significantly associated with tumor size greater than 5cm, mitotic index greater than 5/50 CGA, presence of necrosis, a high risk level, and a cellular proliferation index (Ki67) higher than 5 percent on the reduction of overall survivel of patients (p = 0.017, 0.010, 0.001, 0.016 and 0.0005, respectively). Other factors such as histological subtype, immunophenotype and p16ink4a were not significant. CONCLUSION: According to our data, risk level, tumor size, mitotic index and the presence of necrosis stood as morphological predictors of reduced survival, which underpins previous evidence of their application. The cellular proliferation marker Ki67 associated with risk level also proved to be a useful predictor of tumor aggressiveness.

Dendritic right/left asymmetries in the neurons of the human hippocampal formation: a quantitative Golgi study

OBJECTIVE: To search for right/left asymmetries in the dendritic trees of the neuronal populations and in the cell-free layer volumes of the human hipoccampal formation. METHOD: In necropsic material obtained from six male individuals we performed a quantitative Golgi study of the dendritic trees of dentate granules, CA3 and CA1 pyramidal neurons and a volumetric analysis of dentate gyrus molecular layer, strata oriens plus alveus and strata lacunosum-moleculare plus radiatum of CA3 and CA1 fields. RESULTS: We found inter-hemispheric asymmetries in the dendrites trees of all neurons, reaching the significant level in the number of granule cells dendritic segments (higher in the left than in the right hemisphere), dendritic branching density of CA3 pyramidal cells and mean dendritic length of CA1 apical terminal segments (higher in the right than in the opposite side). No volumetric differences were observed. CONCLUSION: This study points to different anatomical patterns of connectivity in the hippocampal formations of both hemispheres which may underlie functional asymmetries.

OBJETIVO: Pesquisar a existência de assimetrias direita/esquerda nas arborizações dendríticas neuronais e nos volumes das camadas não celulares da formação do hipocampo humano. MÉTODO: Efectuamos estudo quantitativo Golgi das arborizações dendríticas dos grânulos da fascia denteada e das células piramidais de CA3 e CA1, e uma análise estereológica dos volumes da camada molecular da fascia denteada, do strata oriens + alveus e do strata lacunosum-moleculare + radiatum de CA3 e de CA1 em material necrópsico colhido em 6 indivíduos do sexo masculino. RESULTADOS: Encontrámos assimetrias inter-hemisféricas nas arborizações dendríticas de todos os neurónios, significativas no número de segmentos dendríticos das células granulares (maior à esquerda do que à direita) na densidade de ramificação dendrítica das pirâmides de CA3 e no comprimento dendrítico médio dos segmentos apicais terminais das pirâmides de CA1 (maiores à direita do que à esquerda). Não encontramos diferenças volumétricas. CONCLUSÃO: Estes resultados alertam para diferentes padrões anatómicos de conectividade nas formações do hipocampo de ambos os hemisférios que podem fundamentar assimetrias funcionais.

Associação entre tamanho e potencial proliferativo em neurinomas do acústico / Size and proliferative index correlation in acoustic neuromas

Schwanomas do acústico são os tumores mais freqüentes localizados no ângulo pontocerebelar. Os mecanismos moleculares que levam a sua geração e crescimento ainda não são bem conhecidos. Várias características clínicas, radiológicas e imuno-histoquímicas já foram estudadas e correlacionadas ao crescimento tumoral. Estudamos e correlacionamos aspectos clínicos e imuno-histoquímicos (MIB-1) de 11 schwanomas do acústico operados no Hospital São Paulo/UNIFESP. O tamanho dos tumores correlacionou-se com o índice proliferativo (Ki-67), não havendo correlação com significância estatística entre a idade dos pacientes, duração dos sintomas e índice proliferativo.

Morphometric study of the postnatal growth of the parotid gland of the mouse

O crescimento das glândulas parótidas do camundongo durante 7 e 35 dias de vida pós-natal foi estudado por métodos morfométricos. A massa glandular, o volume de cada compartimento morfológico e o número de células em cada compartimento foram avaliados. Os dados obtidos para cada dimensão avaliada foram ajustados por equação exponencial, tipo Y = a.ek.x, permitindo o cálculo do seu tempo de duplicação médio (TD), ou seja, uma estimativa da sua velocidade de crescimento. A análise dos resultados mostrou a ocorrência de um marcante aumento de massa glandular no período estudado de 1.424%, com TD = 7,10 dias. Esse crescimento glandular ocorreu por aumentos nos volumes absolutos dos ácinos, dos ductos intercalares, dos ductos estriados, dos ductos excretores e do estroma, com aumentos percentuais de, respectivamente, 3.048%, 417%, 2.662%, 2.594% e 367%, e TDs de 5,62, 11,71, 5,55, 5,47 e 14,45 dias. A análise da evolução do número de células em cada compartimento demonstrou aumentos de, respectivamente, 1.904%, 285%, 1.228%, 1.090% e 286% e TDs de 6,62, 20,40, 7,19, 7,26 e 14,51 dias. Baseados nos resultados aqui obtidos, concluímos que o crescimento das glândulas parótidas do camundongo entre os dias 7 e 35 de idade ocorre por intenso acúmulo de células, principalmente nos ácinos e nos ductos estriados e excretores, com uma velocidade de crescimento sensivelmente maior que nos ductos intercalares e no estroma.

Density, proportion, and dendritic coverage of retinal ganglion cells of the common marmoset (Callithrix jacchus jacchus)

We performed a quantitative analysis of M and P cell mosaics of the common-marmoset retina. Ganglion cells were labeled retrogradely from optic nerve deposits of Biocytin. The labeling was visualized using horseradish peroxidase (HRP) histochemistry and 3-3'diaminobenzidine as chromogen. M and P cells were morphologically similar to those found in Old- and New-World primates. Measurements were performed on well-stained cells from 4 retinas of different animals. We analyzed separate mosaics for inner and outer M and P cells at increasing distances from the fovea (2.5-9 mm of eccentricity) to estimate cell density, proportion, and dendritic coverage. M cell density decreased towards the retinal periphery in all quadrants. M cell density was higher in the nasal quadrant than in other retinal regions at similar eccentricities, reaching about 740 cells/mm² at 2.5 mm of temporal eccentricity, and representing 8-14 percent of all ganglion cells. P cell density increased from peripheral to more central regions, reaching about 5540 cells/mm² at 2.5 mm of temporal eccentricity. P cells represented a smaller proportion of all ganglion cells in the nasal quadrant than in other quadrants, and their numbers increased towards central retinal regions. The M cell coverage factor ranged from 5 to 12 and the P cell coverage factor ranged from 1 to 3 in the nasal quadrant and from 5 to 12 in the other quadrants. These results show that central and peripheral retinal regions differ in terms of cell class proportions and dendritic coverage, and their properties do not result from simply scaling down cell density. Therefore, differences in functional properties between central and peripheral vision should take these distinct regional retinal characteristics into account.

Liquid-base cytology: a new method for oestral cycle study in wistar's rats

OBJETIVOS: O objetivo deste estudo foi à padronização de uma técnica de coleta e coloração em meio líquido que alie a praticidade e a riqueza citológica, possibilitando uma maior reprodutividade e facilidade microscópica. MÉTODOS: Ratas wistar (n=20) foram submetidas à coleta vaginal diária em salina e o lavado fixado (éter/álcool) e corado em suspensão com solução de azul de Evans 0,025 por cento. A amostra foi concentrada por centrifugação e observado sob objetiva de 40 x. RESULTADOS: Os esfregaços corados permitiram nítida diferenciação das fases do ciclo hormonal (diestro, proestro, estro e metaestro); além da diferenciação dos tipos celulares em relação ao seu grau de maturação tendo como parâmetros o tamanho celular, relação núcleo / citoplasma (RNC) e reação tintorial. O estudo demonstrou a existência de três padrões celulares básicos: células com baixa RNC, acentuada cianofília e pequeno tamanho; células com acréscimo na RNC, perda de cianofilia e maior volume citoplasmático e células queratinizadas anucleadas em aspecto de escama. CONCLUSÃO: A coloração do material permitiu, além da classificação citológica, a possibilidade de quantificação o que resultaria em um acompanhamento mais acurado do ciclo estral.

Tissue alterations in the Guinea pig lateral prostate following antiandrogen flutamide therapy

The flutamide antiandrogenic effects on the Guinea pig male prostate morphology in puberal, post-puberal and adult ages were evaluated in the present study. Daily-treated group animals received flutamide subcutaneous injection at a dose of 10 mg/Kg body weight for 10 days. The control group animals received a pharmacological vehicle under the same conditions. The lateral prostate was removed, fixed and processed for light and transmission electron microscopy. The results revealed an increase of the acinus diameter in the treated puberal animals and straitness in the stromal compartment around the acini. The epithelial cells exhibited cubic phenotype. In the post-puberal and adult animals, a decrease of the acinus diameter was observed, as well as an increase of the smooth muscle layer and presence of the folds at epithelium. The ultrastructural evaluation of the secretory cells in the treated group demonstrated endomembrane enlargement, mainly in the rough endoplasmic reticulum and Golgi apparatus. In addition, a decrease of the microvilli and alterations in the distribution patterns and density of the stromal fibrillar components were observed. In conclusion, the flutamide treatment exerts tissue effects on the lateral prostate, promoting stroma/epithelium alterations.

Morphological study of the spermatogenesis in the teleost Piaractus mesopotamicus

The spermatogenesis of Piaractus mesopotamicus was investigated under light and transmission electron microscopy. The specimens were captured from their natural environment (Rio Miranda and Rio Aquidauana, Pantanal Matogrossense, Brazil) during April and September. The results were compared with the spermatogenic data of specimens under captivity condition. In both conditions, P. mesopotamicus presented the typical spermatogenesis pattern of the teleost fishes, showing no significative differences. The spermatozoon was classified as type I, which has a globular head without acrosome, a short middle piece and a long tail constituted only by the flagellum. This type of spermatozoon is considered the basic type in fishes.

Stereological tools in biomedical research

Stereological studies are more and more frequent in literature, particularly in the development/evolution, pathology, and neurosciences areas. The stereology challenge is to understand the structural inner threedimensional arrangement based on the analysis of the structure slices only showing two-dimensional information. Cavalieri and Scherle's methods to estimate volume, and Buffon's needle problem, are commented in the stereological context. A group of actions is needed to appropriately quantify morphological structures (unbiased and reproducibly), e.g. sampling, isotropic and uniform randomly sections (Delesse's principle), and updated stereological tools (disector, fractionator, nucleator, etc). Through the correct stereology use, a quantitative study with little effort could be performed: efficiency in stereology means a minimum slices sample counting (little work), low cost (slices preparation), but good accuracy. In the present text, a short review of the main stereological tools is done as a background basis to non-expert scientists

Changes in cell shape, cytoskeletal proteins and adhesion sites of cultured cells after extracellular Ca2+ chelation

Although much is known about the molecules involved in extracellular Ca2+ regulation, the relationship of the ion with overall cell morphology is not understood. The objective of the present study was to determine the effect of the Ca2+ chelator EGTA on the major cytoskeleton components, at integrin-containing adhesion sites, and their consequences on cell shape. Control mouse cell line C2C12 has a well-spread morphology with long stress fibers running in many different directions, as detected by fluorescence microscopy using rhodamine-phalloidin. In contrast, cells treated with EGTA (1.75 mM in culture medium) for 24 h became bipolar and showed less stress fibers running in one major direction. The adhesion plaque protein alpha5-integrin was detected by immunofluorescence microscopy at fibrillar adhesion sites in both control and treated cells, whereas a dense labeling was seen only inside treated cells. Microtubules shifted from a radial arrangement in control cells to a longitudinal distribution in EGTA-treated cells, as analyzed by immunofluorescence microscopy. Desmin intermediate filaments were detected by immunofluorescence microscopy in a fragmented network dispersed within the entire cytoplasm in EGTA-treated cells, whereas a dense network was seen in the whole cytoplasm of control cells. The present results suggest that the role of extracellular Ca2+ in the regulation of C2C12 cell shape can be mediated by actin-containing stress fibers and microtubules and by intermediate filament reorganization, which may involve integrin adhesion sites