The cytogenetic examination as a tool for the diagnosis of chromosomal disorders / Examinación citogenética como herramienta para el diagnóstico de los desórdenes cromosómicos

Clinically significant chromosomal abnormalities occur in about 1 percent of children born alive. The objective of this work was to offer the patients and the families in the community for the service of the Integrated Clinic of Uniara Health (Araraquara and region), the examination of cariotype (cytogenetic study) for confirmation or exclusion of the diagnostic suspicion of chromosomal abnormalities as well as information (genetic counseling) for the prevention of occurrence and/or recurrence of these anomalies. In the period of one year and four months these were carried out in the Integrated Clinic of Uniara Health and directed for the Laboratory of Cytogenetic Human of the same institution in 66 cytogenetic studies. In 44 patients (66.6 percent) the results were normal. In 22 (33.3 percent) examinations, alterations were found, meaning that the respective clinical pictures are decurrent of chromosomic alterations. The first cause within alterations noted was Down syndrome with a total of 15 examinations or 68.1 percent, the second cause of chromosomal anomaly was the Turner syndrome where the most important factor is 45, X, where 2 karyotypes of this type or 9.1 percent were found, syndromes as (Eduards syndrome, Patau syndrome, 3p- syndrome, 4p- syndrome and 6p-syndrome) diagnosed in our laboratory appeared less frequently corresponding to 22.7 percent of the studied anomalies. The work carried out constitutes a necessary diagnosis of the main chromosomal abnormalities through a low cost technique; it can be carried out easily and is reliable, making the cytogenetic examination available to the community and contributing significantly to the quality of life of patients.

Las anormalidades cromosómicas, clínicamente significativas, se presentan en aproximadamente 1 por ciento de los niños nacidos vivos. Este trabajo tiene el objetivo de ofrecer a los pacientes y /o a sus familiares el servicio de la Clínica Integrada de la Salud de Uniara (Araraquara y Región), el examen de cariotipo (estudio citogenético) para la confirmación o la exclusión de sospecha de anomalías cromosomales diagnosticadas, así como otorgar información (consejo genético) para la prevención de las posibles anomalías y /o la repetición de éstas. En un año y cuatro meses fueron realizados 66 estudios de citogenética en la Clínica Integrada de Uniara, dirigida por el Laboratorio de Citogenética Humana de la misma institución. En 44 pacientes (66,6 por ciento) los resultados fueron normales. En 22 (33,3 por ciento) de los exámenes, se encontraron alteraciones, compatibles con alteraciones cromosómicas. La primera causa de anomalías cromosómica fue el síndrome de Down, totalizando 15 exámenes (68,1 por ciento), la segunda causa fue el síndrome de Turner, con dos cariotipos (9,1 por ciento) en la forma más importante 45, X. Por otra parte, se encontró que los síndromes de Eduards, de Patau, 3p-síndrome de Down, síndrome 4p-6p, diagnosticados en nuestro laboratorio, presentaban baja frecuencia de aparición, representando el 22,7 por ciento de las anomalías estudiadas. Este trabajo permitió realizar un diagnóstico preciso de las anomalías cromosomales, principalmente a través de una técnica de bajo costo, fácil ejecución y buena confiabilidad, técnicas que están disponibles para el examen citogenético para la comunidad y así contribuir de manera significativa en la calidad de vida de los pacientes.

A statistical study of the association of seven dental anomalies in the Brazilian population / Estudio estadístico de la asociación de siete anomalías dentales en la población brasileña

The objective of this study was to show the association patterns among seven types of dental anomalies (second pre-molar agenesis, upper side incisive reduced in size, lower first molar infra-ochlesis, enamel hypoplasia, first molar ectopic eruption, supra numerous teeth and upper canine ectopic eruption) in a population sample without dental treatment ranging in age from 7 to 14. A total of 172 patients were attended and underwent the clinical examination at the Clínica Infantil da Fundação Educacional de Barretas. Eleven patients from this total were selected according to a first dental anomaly diagnosis and submitted to panoramic radiography. A significant association (p<0.05) was detected among six pairs of anomalies (second pre-molar agenesis x first pre-molar ectopic eruption; second pre-molar agenesis x lower first molar infra-ochlesis; second pre-molar agenesis x upper side incisive reduced in size; supra numerous teeth x reduced size upper side incisive; first pre-molar ectopic eruption x enamel hypoplasia; lower first molar infra-ochlesis x upper side incisive reduced in size) suggesting a common genetic origin for these conditions. The association was not significant in only one case where there was anomaly sharing by the patients. The existence of an anomaly is clinically relevant for early diagnosis of a possible association and an anomaly can indicate an increased risk of other anomalies.

El objetivo de este estudio fue mostrar los patrones asociación entre siete tipos de anomalías dentales (agenesia del segundo premolar, incisivo lateral superior en tamaño reducido, infra-oclesis del primer molar inferior, hipoplasia del esmalte, erupción ectópica del primer molar, dientes supernumerarios y erupción ectópica de caninos superiores) en una muestra de población sin tratamiento dental, de edades comprendidas entre los 7 a 14 años. Un total de 172 pacientes fueron atendidos y se les realizó el examen clínico en la Clínica Infantil da Fundación Educacional de Barretos. Once pacientes de el total fueron seleccionados de acuerdo con un primer diagnóstico de anomalías dentales y presentado en la radiografía panorámica. Se observó una asociación significativa (p <0,05) entre los seis pares de anomalías (agenesia de segundo premolar x erupción ectópica del primer molar; agenesia del segundo premolar x infra-oclesis del primer molar inferior; agenesia del segundo premolar x incisivo lateral superior en tamaño reducido; dientes supernumerarios x incisivo lateral superior en tamaño reducido; erupción ectópica del primer molar x hipoplasia de esmalte; infra-oclesis del primer molar inferior x incisivo lateral superior en tamaño reducido), sugiriendo un origen genético común para estas condiciones. La asociación no fue significativa en un sólo caso donde hubo anomalías compartidas por los pacientes. La existencia de una anomalía es clínicamente relevante para el diagnóstico precoz de una posible asociación y una anomalía puede indicar un mayor riesgo de otras anomalías.

Morphometric Analysis of Nucleus and Nucleolar Organizer Regions (NORs) in Tongue Squamous Cell Carcinoma (SCC) / Análisis Morfométrico de Núcleos y Regiones del Organizador Nucleolar (NORs) en Carcinomas de Células Escamosas de la Lengua (SCC)

The tongue is one of the most likely places for the primary squamous cell carcinoma (SCC) to arise in the western world.This work intends to do a morphometric evaluation of the cellular nucleus, as well as a morphometric analysis of the nucleolar organizer regions (NORs). The biopsy specimen was removed from patients with tongue SCC, staged T2N0M0, before they underwent treatment. They were, then, subject to hemiglossectomy and supraomohyoid neck dissection (SOHND) and presented different clinical evolution. According to that, they were subdivided in three groups: Group A, control group, formed by patients without tongue neoplasm (the histopathologic specimen was obtained from the Department of Pathology at Hospital de Base and Faculdade de Medicina de Sao José do Rio Preto - FAMERP-HB); Group B, T2N0M0+ hemiglossectomy + SOHND, survival of only 36 months; Group C, T2N0M0 + hemiglossectomy + SOHND, survival of 5 years. The morphometric analysis included the study of larger diameter, smaller diameter, medium diameter, the relation between larger/smaller diameters, perimeter, area, volume, the relation between volume/area, idiosyncrasy, shape coefficient and the nuclear contour index of the tumor cells from patients with tongue SCC, as well as the same measurements of the nucleolar organizer regions. The analysis was aleatory and double-blind. All the morphometric data were compiled and statistically evaluated by the Kruskall-Wallis test and by the ANOVA test, and a 5% alpha error rate was adopted.

La lengua es probablemente, el lugar en donde más frecuentemente surge el carcinoma primario de células escamosas (SCC), en el mundo occidental. Este estudio propone realizar la evaluación morfométrica del núcleo celular, así como el análisis morfométrico de las regiones de los organizadores nucleolares (NORs). La muestra de biopsia fue removida de pacientes con SCC de lengua, etapificado en T2N0M0, antes de recibir tratamiento. Los pacientes fueron sometidos a una hemiglosectomía y una disección supraomohiodea del cuello (SOHND) y presentaron diferente evolución clínica. De acuerdo a esto, fueron subdividos en tres grupos: Grupo A, grupo control formado por pacientes sin neoplasia de lengua (la muestra fue obtenida del Departamento de Patología del Hospital Base de la Facultad de Medicina de Sao José do Rio Preto - FAMERP-HB, Brasil); Grupo B, T2N0M0 + hemiglosectomía + SOHND, sobrevida de 36 meses; Grupo C, T2N0M0 + hemiglosectomía + SOHND, sobrevida 5 años. El estudio morfométrico incluyó los análisis del diámetro mayor, diámetro menor, diámetro medio, la relación entre el diámetro mayor/menor, perímetro, área, volumen, relación volumen/área, idiosincrasia, coeficiente de forma y el índice de contorno nuclear de células tumorales en pacientes con SCC de lengua, así también las mismas mediciones de las regiones nucleolares organizadas. El análisis fue aleatorio y doble ciego. Todos los datos morfométricos fueron recopilados y evaluados estadísticamente con el test de Kruskall-Wallis y el Test ANO VA, con un valor alfa de un 5%.

Cyclophosphamide Effect on the Epithelial Covering of Rats Fetuse's Tongue: A Morphometric Study / Efectos de la Ciclofosfamida en el Epitelio de Revestimiento de la Lengua de Fetos de Rata: Un Estudio Morfométrico

The present study has the aim to describe morphologically, morphometrically and stereologically some changes in the tongue epithelium of rat fetuses under the effect of cyclosphosphamide. This drug was used in a single 6mg/kg according to the body weight in the tenth ­ pregnancy day while the control group received equal dosage of saline solution by the same duct and in the same day of pregnancy. The fetuses were plunged into fixed solution of Bouin for five days and the heads of then were divided and blocked in paraffin. Histological sections of six-micrometer thickness were performed and then colored with hematoxilin and eosin. It was used morphometrical and stereological analysis after the morphological study. The data were submitted under the statistical non-parametric test of Mann-Whitney. The results showed that the cyclosposphamide usage effect during the rat pregnancy caused a delayed in the embryo fetal growing, a reduction in the placental weight as wells as in the umbilicus cord length. The covering epithelia in the tongue of the treated group showed some changes since there was an epithelial hyperplasia associated with a cell hypotrophy. The fetuses in this group were more premature than the ones in the control.

El estudio tiene como objetivo describir morfológica, morfométrica y estereológicamente algunos cambios en el epitelio de la lengua de fetos de rata bajo los efectos de la ciclofosfamida. Esta droga fue usada en una muestra única de 6mg/kg de acuerdo al peso del cuerpo, en el décimo día de preñez, mientras el grupo control recibió igual dosis de una solución salina, por la misma vía, en los mismos días de preñez. Los fetos fueron sumergidos en solución fijadora de Bouin, por cinco días y las cabezas de ellos fueron cortadas y colocadas en bloques de parafina. Secciones histológicas de 6 µm fueron teñidas con hematoxilina-eosina. Después del estudio morfológico se realizaron análisis morfométricos y estereológicos. Se utilizó el test estadístico no paramétrico de Mann-Whitney. Los resultados mostraron que el uso de la ciclofosfamida provoca efectos durante la preñez de la rata, causando un retardo en el crecimiento de los embriones, una reducción en el peso de las placentas como también, en la longitud del funículo umbilical. El epitelio de revestimiento de la lengua del grupo tratado, mostró algunos cambios desde una hiperplasia epitelial asociada con una hipotrofia celular. Los fetos del grupo tratado fueron más prematuros que los del grupo control.

Renal morphometry of fetuses rats treated with cadmium

El cadmio es un metal pesado que se encuentra en la tierra, aire y alimentos. Está presente en forma coditiana todos los días de la vida y actualmente es catalogado como un tóxico, teratogénico y agente carcinogénico. El cadamio causa numerosas lesiones a los órganos humanos y animales. Este trabajo tiene como objetivo investigar los efectos tóxicos en fetos de ratas expuestas a tratamiento con cloruro de cadmio. 10 ratas pre¤adas fueron divididas en dos grupos. Cinco ratas recibieron por administración intraperitoneal 5 mg/Kg de cloruro de cadmio por peso corporal, durante 10 días de pre¤ez y otras cinco ratas recibieron bajo las mismas condiciones del otro grupo, una solución de cloruro de sodio al 0.85 por ciento . Los fetos fueron extraídos junto con las placentas y funículos umbilicales a los 20 días de pre¤ez. La administración del cloruro de cadmio causó una reducción significativa en los pesos de los fetos y placentas. La longitud del funículo umbilical fue más corta en el grupo experimental que en el grupo control. El estudio evaluó las alteraciones morfológicas causadas en los túbulos proximal, distal, contorneados y colectores renales como también en las células del glomérulo renal. La morfometría evidenció alteraciones significativas en los túbulos proximales y en las células glomerulares y, en menor grado, en los túbulos distales. El borde en cepillo de la membrana celular se observó muy sensible y se visualizaron algunas alteraciones. Los glomérulos se presentaron con edema y la cápsula del glomérulo fue también afectada.

Effects of low-dose phenytoin administered to rats fetuses on developing choroid plexus: kariometric study

The aim of the present study was to determine, by kariometric parameters, the influence of phenytoin, as teratogen, on the epithelial layer of choroid plexus in rats fetuses, using single daily dose ( 75 mg Kg-1 bd.wt.), during gestation GD9 to GD11, and lowest total dose used orally (225 mg Kg-1), to the best of our knowledge. In some of experimental studies in animals, the anticonvulsant drug phenytoin were administered in embriotoxic daily doses (100-1500 mg Kg-1 bd.wt.) increasing concentrations up to 20 times the human therapeutic plasma concentration. There is a significant lack of information regarding the embriotoxicity in pregnancy of these drug, phenytoin, in low doses. Phenytoin was administered in a single daily dose (75 mg Kg-1 bd.wt.) to pregnanty rats in GD9 to GD11 days of gestation, during organogenesis. Histological sections were obtained for analysis of nuclear alterations in the cuboidal epithelium of choroid plexuses of lateral ventricles in the rat fetuses. Eleven kariometric parameters were measured in each of the nuclei. Statistical comparison were made with Mann-Whitney's test. The nuclei of the phenytoin group showed significant reduction of the size parameters : longest axis, mean axis, nuclear volume, nuclear area, nuclear perimeter, ratio of the longest axis to the shortest axis, ratio of the nuclear volume to the nucleararea) but not in shortest axis and shape parameters (contour index, shape factor and eccentricity) . A distinctive pattern of nuclear abnormalities in choroid plexus ephitelium of rat fetuses is associated with the use of low dose of phenytoin during pregnancy. Variations in nuclear size might reflect fundamental nuclear alterations of significance during the process of embriogenesis and could represent teratogenic influence of phenytoin in rats. Even at low dosage and short period of use during gestation, phenytoin can induce embriotoxicity.

Teratogenic effects of lamotrigine on rat fetal brain: a morphometric study.

A study of the teratogenic activity of an antiepileptic drug - lamotrigine - was carried out in the brain of fetuses of rats who had received the drug. The dosage levels studied corresponded to four times the median effective dose (ED50) in rats. The drug was administered during the organogenesis period. Rats were sacrificed one day prior to term and fetuses were macroscopically examined, weighted and cephalic segments sectioned (Wilson technique), for histological study by stereological analysis, using Merz's grid for drawing and point counts. Cortex, subcortex, ependyma and lateral ventricles were analyzed. The same methodology was applied to the control group; data were compared with by the non-parametric Mann-Whitney statistical analysis test. Results showed that fetuses of the experimental group had reduced body weight at birth, increased volume and diameter of the cerebral structure, increased density of the subcortical layer, and ventricle dilatation. Possible mechanisms of this teratogenicity were discussed.

Effect of acyclovir on rat fetus palate mucosa.

Five pregnant rats were treated during organogenesis with sc injections of acyclovir (50 mg/kg body weight) on days 9, 10 and 11 of pregnancy. The fetuses (N=62) were evaluated on day 20 of gestation and presented decreased body weight as well as delayed differentiation of fetal rat palate epithelium, with increased nuclear volume, decreased cytoplasmic and cellular volumes, decreased epithelial and keratin thicknesses, and increased cellular numerical density.

Inseminaçäo tubária indireta com alto volume de esperma humano capacitado (ITI) / Indirect fallopian insemination with high volume of washed human sperm

Os autores descrevem uma técnica para inseminaçäo tubária indireta com alto volume de esperma humano capacitado injetado através de uma cânula especial. O procedimento foi aplicado para o tratamento de infertilidade em sete mulheres com no mínimo uma trompa permeável na histerossalpingografia e resultou em duas gestaçöes normais em evoluçäo

Análise morfométrica do testículo e epididimo do rato na fase inicial da intoxicaçäo pelo chumbo

Ratos machos adultos foram intoxicados por ingestäo "ad libitum" de acetato de chumbo nas concentraçöes de 0,5 g/1 e 1,0 g/l, durante 90 dias. Ao final do período experimental, a intoxicaçäo foi confirmada pelo aumento acentuado na concentraçäo de chumbo no sangue, aliado `a diminuiçäo dos valores de hematócrito e hemoglobina e aumento da glicemia. Nesta fase inicial da intoxicaçäo, näo se observaram alteraçöes no diâmetro dos túbulos seminíferos, na altura do epitélio germinativo, no ritmo do processo espermatogênico e na produçäo de espermatozóides. A cabeça do epidídimo näo apresentou também qualquer alteraçäo estrutural. A cauda do epidídimo, por outro lado, exibiu aumento significativo no diâmetro do ducto e diminuiçäo na altura do epitélio. A concentraçäo de espermatozóides armazenados na regiäo caudal do epidídimo mostrou-se significativamente aumentada nos aniamis tratados com chumbo. Os resultados revelam o envolvimento precoce da cauda do epidídimo na intoxicaçäo pelo chumbo, e sugerem um distúrbio no mecanismo neuroendócrino que controla as múltiplas funçöes epididimárias

Functional disturbance of rat sexual accessory glands in an early phase of lead intoxication.

In order to investigate the effects of chronic lead intoxication on sexual accessory glands adult male Wistar rats were poisoned by ad libitum ingestion of lead acetate at concentrations of 0.5 g/l and 1.0 g/l for 90 d. Blood lead exhibited a significant increase in both treated groups. A decrease in hematocrit and hemoglobin, besides a rise in glycemic levels, confirmed lead intoxication. No sign of lesion was detected upon histologic examination of prostate, seminal vesicle, and coagulating gland. A morphometric technique revealed that, in this early phase of intoxication, no alteration occurred in the prostate's and seminal vesicle's epithelial height. However, a significant decrease in fructose content was observed in both lead treated groups. The results, revealing a precocious envolvement of male sexual accessory glands in lead intoxication, were related to disturbance in plasma testosterone level and also probably in androgen receptors.

Morphometric and stereological analysis of rat testis and epididymis in an early phase of saturnism.

Adult male Wistar rats were poisoned by ad libitum ingestion of lead acetate at concentrations of 0.5 g/l and 1.0 g/l in drinking water, for 90 days. Blood lead exhibited a significant increase in both treated groups. A decrease in haematocrit and haemoglobin, together with a rise in glucose levels, confirmed lead intoxication. No signs of lesion were detected upon histological examination of testes, caput and cauda epididymidis. In this early phase of intoxication, no alteration occurred in the seminiferous tubule diameter, in the germinal epithelium height, nor in the rate of spermatogenesis and the production of spermatozoa. The caput epididymidis also showed no structural change. In the cauda epididymidis, however, an increase in ductal diameter, and a decrease in epithelial height, were observed. The concentration of spermatozoa stored in the caudal region of the epididymis exhibited a significant increase in lead-treated animals. The results, which reveal an early involvement of cauda epididymidis in lead intoxication, are discussed in terms of a disturbance in the neuroendocrine mechanism controlling the multiple epididymal functions.

Ulcera de Marjolin: relato de um caso com período de latência de 70 anos / Marjolin's ulcer: case report with lag-time of 70 years

Apresenta-se um caso de úlcera de Marjolin, desenvolvido sobre cicatriz de queimadura, com tempo de latência de 70 anos. Em revisäo de literatura realizada, foi encontrado apenas un caso descrito com tempo de latência maior que 70 anos

Dose-dependent induction of embryonic abnormalities in vitro by tissue homogenates of placenta and decidua.

Homogenate preparations from normal rat placental and decidual tissue induced abnormalities when included in the culture medium of rat embryos between Day 9.5 and Day 11.5. Abnormal embryos were produced between doses of 2.5 and 4 mg/ml for the placental homogenate and between doses of 1.2 and 4 mg/ml for the decidual homogenate, but were not produced by a solution of bovine serum albumin or by a protein preparation of rat lung tissue at the same concentration. The degree to which the embryos were malformed depended on the dose and which of the two homogenates was used. The decidual homogenate preparation was more pathogenic than the placental homogenate, but both were able to produce neural-tube defects and a severe reduction in embryonic size. The possible association between these findings and some known proteins within such homogenates is discussed.

Morphometric study of malformations induced by hypervitaminosis A on rat fetus Meckel cartilage, maxillary bone and mandible.

Morphological and histological studies were carried out to determine the effects on fetal development in rats of a single intraperitoneal injection of 150,000 I.U. of vitamin A, given to the mothers on the 10th day of gestation. The Meckel cartilage of malformed animals appeared to be diminished, with larger nuclei, less numerous isogenous nests, smaller cellular and thin perichondrium and more abundant cartilaginous matrix, which was less deeply stained by alcian blue. The maxillary bone and mandible of the treated fetuses showed thin and poorly calcified trabeculae, smaller medullary spaces and a less fibrous, and more cellular periosteum. These data were confirmed with morphometric techniques.

Morphometric study of the reversibility of testicle alterations in rats submitted to hypervitaminosis A.

In a previous paper, Lamano Carvalho et al. [4] demonstrated that administration of excess vitamin A doses to adult rats causes decreased testicle mass, focal lesions in the seminal tubule epithelium, and reduction in the relative volume of interstitial tissue and in the nuclear volumes of Leydig cells. The hypervitaminotic animals also showed alterations in the rhythm of spermatogenesis and reduced hypophysary LH levels. In order to determine the possible reversibility of these alterations, the authors allowed the hypervitaminotic animals to recover for 50 or 100 days. After both recovery periods. the focal lesions in the germinal epithelium were still unreversed. On the other hand, the nuclear volume of Leydig cells and the rhythm of spermatogenesis, as well as hypophysary LH levels reverted to normal during the 50 day recovery period.

Morphometric study of testicle alterations in rats submitted to hypervitaminosis A.

Testicle lesions and spermatogenetic activity in adult rats submitted to hypervitaminosis A were determined by quantitative studies of the germinal epithelium and of Leydig cells. Hypophysary levels of luteinizing hormone (LH) and relative weights of the adrenals were also determined, and the results were interpreted as being caused by the double mechanism of action (direct and indirect) of vitamin A.