Osteogenesis imperfecta: Ultrastructural and histological findings on examination of skin revealing novel insights into genotype-phenotype correlation.

Osteogenesis imperfecta (OI) is a heterogeneous group of inherited disorders of bone formation, resulting in low bone mass and an increased propensity to fracture. Over 90% of patients with OI have a mutation in COL1A1/COL1A2, which shows an autosomal dominant pattern of inheritance. In-depth phenotyping and in particular, studies involving manifestations in the skin connective tissue have not previously been undertaken in OI. The aims of the study were to perform histological and ultrastructural examination of skin biopsies in a cohort of patients with OI; to identify common and distinguishing features in order to inform genotype-phenotype correlation; and to identify common and distinguishing features between the different subtypes of OI. As part of the RUDY (Rare Diseases in Bone, Joints and/or Blood Vessels) study, in collaboration with the NIHR Rare Diseases Translational Research Collaboration, we undertook a national study of skin biopsies in patients with OI. We studied the manifestations in the skin connective tissue and undertook in-depth clinical and molecular phenotyping of 16 patients with OI. We recruited 16 patients: analyses have shown that in type 1 collagen mutation positive patients (COL1A1/ COL1A2) (n-4/16) consistent findings included: variable collagen fibril diameter (CFD) and presence of collagen flowers. Histological examination in these patients showed an increase in elastic fibers that are frequently fragmented and clumped. These observations provide evidence that collagen flowers and CFD variability are consistent features in OI due to type 1 collagen defects and reinforce the need for accurate phenotyping in conjunction with genomic analyses.

Pregnancy outcome in ethanol-treated mice with folic acid supplementation in saccharose.

INTRODUCTION: Maternal folic acid deficiency is the most important metabolic factor in the etiology of neural tube defects (NTD) and is reduced by ethanol, which is extensively consumed by young women. OBJECTIVE: The objective of the study was to determine whether folic acid supplementation in dietary saccharose is efficient in the prevention NTD induced by ethanol in fetuses of Swiss mice. MATERIALS AND METHODS: Pregnant mice were divided into four groups of six animals each: control (C), ethanol (E), deficient-supplemented (DS), and deficient-supplemented + ethanol (DSE). Groups C and E received commercial mouse chow (containing 3 mg/kg folic acid) throughout the experiment, while groups DS and DSE received a folic acid-free diet with the addition of saccharose supplemented with folic acid (2 mg/kg folic acid) in water. Group E and DSE animals received ethanol (4 g/kg) administered intraperitoneally from the seventh to the ninth gestational day (gd) and were euthanized on the 18th gd, while groups C and DS received saline. RESULTS: Congenital anomalies were observed in groups E and DSE. The fetal weight and length of the animals in group E were lower than in groups C and DS and, in group DSE, were lower than in groups C and DS. The placental diameter of group E was smaller than that of group C, and the placental weight of group C animals was lower than that of groups E, DSE, and DS. CONCLUSION: The study demonstrated that dietary supplementation with folate in saccharose is an accessible means of consumption that could be further diffused but in an increased dose than recommended to reduce the teratogenic effects of ethanol.

Technologies for fixed-time artificial insemination and their influence on reproductive performance of Bos indicus cattle.

The adaptation of Bos indicus cattle to tropical and subtropical environments has led to their widespread distribution around the world. Although artificial insemination (AI) is one of the best alternatives to introduce new genetics into Bos indicus herds, the peculiarity of their temperament and the tendency to show short oestrus (many of them during the night) greatly affects the effectiveness of genetic improvement programs. Therefore, the most useful alternative to increase the number of females that are inseminated is the use of protocols that allow for AI without the need for oestrus detection, usually called fixed-time AI (FTAI). Besides, the development of protocols to advance the resumption of cyclicity during the early postpartum period has a great impact on beef production and will allow for the inclusion of a significantly larger population of animals into genetic improvement programs. Fixed-time AI protocols using progestin devices, oestradiol and eCG have resulted in consistent pregnancy rates in suckled Bos indicus and Bos indicus x Bos taurus cows. Furthermore, fertility in the successive cycles and the overall pregnancy rates at the end of the breeding season, have been shown to be improved by the use of progestin devices at the beginning of the breeding season. In summary, exogenous control of luteal and follicular development has facilitated the application of assisted reproductive technologies in Bos indicus-influenced cattle, by offering the possibility of planning programs without the necessity of oestrus detection and may provide the opportunity to improve reproductive performance of beef cattle in tropical climates.

P63 expression in hydropic abortion and gestational trophoblastic diseases.

Gestational trophoblastic diseases are a group of interrelated diseases of trophoblastic tissue that include partial hydatidiform mole, complete hydatidiform mole, invasive mole, choriocarcinoma, and placental site trophoblastic tumor. P63 is a p53 homologue that, in normal placentas, is expressed in the cytotrophoblast cells. The role of p63 in gestational trophoblastic diseases, however, merits further investigation. Immunohistochemistry with the p63 antibody (clone 4A4) was performed in formalin-fixed paraffin-embedded samples of hydropic abortion (n=10), partial hydatidiform mole (n=12), complete hydatidiform mole (n=12) and choriocarcinoma (n=5). P63 expression was quantitatively assessed as 0 (no stained cells), + (less than 10% positive cells), ++ (10-50% positive cells), and +++ (more than 50% positive cells). The intensity was scored as 0 (absence), + (weak), ++ (moderate), or +++ (strong). Statistical analysis was carried out by the Fisher test. In contrast to the other diagnoses, none of the choriocarcinomas analyzed exhibited p63-positive cells. There was no difference in distribution of p63 positive cells between hydropic abortion, partial hydatidiform mole, and complete hydatidiform mole. Concerning the intensity of immunostaining, there was difference only between partial hydatidiform mole and complete hydatidiform mole. According to our results, p63 might be useful to differentiate a choriocarcinoma from other gestational trophoblastic diseases. Besides, since the intensity of p63 expression was much stronger in partial hydatidiform mole and complete hydatidiform mole than in hydropic abortion, this feature may be helpful in distinguishing these two diagnoses in challenging cases.

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95 percent ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard© genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 æg, with significant differences between fixation methods. Scrape preparation fixed in 95 percent ethanol provided larger amount of extracted DNA. However, the mean for all groups was higher than the quantity needed for PCR (50 ng) or Southern blot (500 ng). There were no qualitative differences among the different material and fixatives. The same results were also obtained for glass slides stored at room temperature for 6, 12, 18 and 24 months. We conclude that touch and scrape preparations fixed in 95 percent ethanol are a good source of DNA and present fewer limitations than cell culture, tissue paraffin embedding or freezing that require sterile material, culture medium, laboratory equipment and trained technicians. In addition, they are more practical and less labor intensive and can be obtained and stored for a long time at low cost.

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells.

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95% ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 microg, with significant differences between fixation methods. Scrape preparation fixed in 95% ethanol provided larger amount of extracted DNA. However, the mean for all groups was higher than the quantity needed for PCR (50 ng) or Southern blot (500 ng). There were no qualitative differences among the different material and fixatives. The same results were also obtained for glass slides stored at room temperature for 6, 12, 18 and 24 months. We conclude that touch and scrape preparations fixed in 95% ethanol are a good source of DNA and present fewer limitations than cell culture, tissue paraffin embedding or freezing that require sterile material, culture medium, laboratory equipment and trained technicians. In addition, they are more practical and less labor intensive and can be obtained and stored for a long time at low cost.

Noonan syndrome associated with unilateral iris coloboma and congenital chylothorax in an infant.

Chylothorax and colobomas are uncommon features reported in Noonan syndrome. We describe an infant with Noonan phenotype, congenital chylothorax and a unilateral iris coloboma. The presence of these both abnormalities in the same patient has not previously been reported.

De novo unbalanced t(11q;21q) leading to a partial monosomy 21pter-q22.2 and 11q24-qter in a patient initially diagnosed as monosomy 21.

We describe a patient in whom full monosomy 21 was initially assumed from routine GTG banded karyotyping. Re-examination with chromosome painting demonstrated an unbalanced translocation between the long arms of chromosomes 11 and 21. Fluorescence in situ hybridization (FISH) and microsatellite marker analysis revealed partial monosomy of chromosome 21 (pter-q22.2) and 11 (q24-qter). The patient was prematurely born in the 31st week of gestation and expired 3 days after delivery. She showed multiple minor anomalies, a complex cardio-vascular malformation, intestinal malrotation and cerebellar hypoplasia.

Raine dysplasia: a Brazilian case with a mild radiological involvement.

We report a preterm male infant, the first child of a young consanguineous couple, whose physical examination revealed craniofacial disproportion with microcephaly, wide fontanelles, exophthalmos, low nasal root and hypoplastic nose, long philtrum, small mouth, high arched and narrow palate, micrognathia, dysplastic, low-set and rounded ears, short neck and, arthrogryposis. Postmortem findings included hypoplastic lungs. Radiological examinations showed mild and localized increased of bone density in the cranial vault and skull base and facial bones and undermodelled in the long bones. The above findings are characteristics of Raine dysplasia but the case reported here presents a mild bone involvement with only a localized bone sclerosis and absence of prenatal fractures. We discuss the possibility that this case represents an allelic mutation of the Raine gene. The consanguinity of the parents reinforces the hypothesis of autosomal recessive inheritance for this entity.

Hepatic capillariasis in children: report of 3 cases in Brazil.

Capillaria hepatica is a helminth that may cause an extremely rare condition of parasitic hepatitis. Only 29 cases have been published, 2 of them in Brazil. We report here 3 cases of children in Brazil with massive hepatic capillariasis who presented the characteristic triad of this type of infection, i.e., persistent fever, hepatomegaly, and eosinophilia. The diagnosis was made by liver biopsy. All children responded well after treatment with thiabendazole (case 1), albendazole (case 3), and albendazole in combination with a corticoid (case 2). Case 1 has been followed-up for 24 years, an event not previously reported in the literature.

[Implantation of a program of terminal evaluation of graduating medicalstudent performance to estimate the efficacy of the curriculum in the ++Ribeirão Preto Medical School]. / Implantação de um programa de avaliação terminal do desempenho dos graduandos para estimar a eficácia do currículo na Faculdade de Medicina de Ribeirão Preto.

PURPOSE: To describe the main steps in the implementation of such assessment, as well as to present the measures taken in order to overcome the difficulties that have been found. METHODS: The proposed two-step assessment consists of a test of cognitive aspects based on Multiple Choice Questions (MCQ) and practical exams of clinical skills in each of 5 terminal areas. Students enrolled as volunteer for either step or both. The examinations were developed and carried out by faculty members nominated by the involved Departments under the supervision of an institutional work group. The rates of student enrollment for the summative assessment has been recorded and the responses of both students and faculty members were evaluated by means of specific questionnaires. RESULTS: In the first two years, adhesion of students to the MCQ test was consistently high, but volunteering to the practical exams was persistently low. Moreover, there was a sharp decrease in faculty involvement, from the first to the second year. To overcome these difficulties, a number of measures were implemented aiming at increasing student adhesion and faculty involvement, as well as to improve assessment methods. As a consequence, a remarkable increase in both student adhesion and faculty involvement was recorded. CONCLUSION: The measures taken resulted in sharp increases in both acceptance of the proposed assessment and the quality of the examination methods, which have allowed a more reliable characterization of the strenghts and weaknesses of the local curriculum.

Implantaçäo de um programa de avaliaçäo terminal do desempenho dos graduados para estimar a eficácia do currículo na Faculdade de Medicina de Ribeiräo Preto / Implantation of program of terminal evaluation of graduating medical students performance to estimate the efficacy of the curriculum in the Faculdade de Medicina de Ribeiräo Preto

Nos últimos cinco anos, um programa de avaliação final do desempenho dos graduandos vem sendo desenvolvido na Faculdade de Medicina de Ribeirão Preto, visando colher dados sobre a eficácia do currículo médico. Objetivo. Descrever as principais etapas de implantação deste programa, bem como as medidas tomadas para superar as dificuldades encontradas. Métodos. A avaliação consiste em uma prova de conhecimento e em exames práticos de habilidades. As provas são elaboradas e aplicadas por docentes das áreas envolvidas sob a supervisão de um grupo coordenador supra-departamental, que avalia todo o programa. Os graduandos participam como voluntários e os resultados são utilizados somente para os fins institucionais propostos. A adesão dos graduandos foi registrada e as impressões dos docentes e dos graduandos sobre o programa foram colhidas por meio de questionários específicos. Resultados. Nos dois primeiros anos a adesão dos graduandos à prova de conhecimento foi superior a 85 por cento, mas o percentual de inscrições nas provas práticas foi de apenas 55 por cento. Além disso, do primeiro para o segundo ano, houve diminuição considerável do grau de empenho dos docentes envolvidos, obrigando medidas corretivas voltadas ao recrutamento dos graduandos, à participação docente e aos métodos de avaliação prática. Em conseqüência, houve aumento notável da adesão dos estudantes que atingiu mais de 90 por cento nos anos seguintes. Houve, também acréscimo considerável do empenho docente, o que garantiu adequado nível de qualidade à avaliação. Conclusão. As medidas tomadas aumentaram a aceitação do programa e a qualidade das provas, permitindo que a avaliação do desempenho dos graduandos traga dados fidedignos sobre a eficácia do currículo.

A viable fetus presenting 68, XX[73]/69, XXX[27] triploid mosaicism

A triploidia é uma anomalia cromossômica comum encontrada em 1 a 2 por cento das gestaçöes clinicamente reconhecidas e em cerca de 15 a 20 por cento dos abortos espontâneos de causa cromossômica. Em aproximadamente 5 por cento dos casos, uma aneuploidia pode estar também associada (Boué et al., 1985). Descrevemos um recém-nascido do sexo feminino, prematuro (30 semanas de idade gestacional), com microcefalia, dismorfias faciais e alteraçöes de membros, que foi a óbito com 1 dia de vida por insuficiência respiratória. O exame anátomo-patológico da placenta revelou alteraçöes compatíveis com degeneraçäo molar. A necrópsia da criança näo evidenciou malformaçöes internas. A análise citogenética de 100 metástases, obtidas a partir de cultura de tecido renal, evidenciou cariótipo 68,XX[73]/69,XXX[27]. Apenas 9 casos de triploidia 68,XX foram descritos anteriormente, sendo 7 em abortos, 1 em feto de 21 semanas e 1 em recém-nascido a termo. Consideramos que este estudo seja o primeiro da literatura relatando a ocorrência de mosaicismo 69,XXX/68,XX em um recém-nascido vivo. Os autores discutem os achados clínicos e os possíveis mecanismos envolvidos nesta aberraçäo cromossômica.

Sudden unexpected infant death syndrome in Ribeirão Preto, Brazil.

CONTEXT: Sudden Infant Death Syndrome (SIDS) is the leading cause of death in the first year of life in developed countries. Brazilian and Latin American literature lack studies on the subject, which is largely unknown among health workers. OBJECTIVE: To identify cases that could be classified as SIDS among children of less than one year of age submitted to autopsy at the Serviço de Verificação de Obitos do Interior (SVOI), in Ribeirão Preto, SP, Brazil. A retrospective analysis of all autopsies from SVOI in this age group from January 1987 to December 1990 was done. RESULTS: There were 369 autopsies of which 344 (93.2%) deaths were expected and 25 (6.8%) unexpected. From the 25 unexpected cases 16 (64%) deaths could not be explained after autopsy and from these cases only 10 were eligible for the study because they had full organ sampling. There were 7 males and 3 females and the age at death ranged from 1 to 3 months (average: 1.7 months). Two were found dead, 3 died at home, 4 died on the way to hospital and 1 died while being fed. Autopsy diagnoses were aspiration (8 cases), SIDS (1 case) and undetermined (1 case). Aspiration was not confirmed by histology and the only findings were mild pulmonary edema, subcapsular petechiae and intraparenchymatous hemorrhage in thymus. CONCLUSION: That there were 10 cases of unexpected and unexplained deaths of children less than 1 year-old during the evaluated period with characteristics similar to SIDS which should therefore be classified as such.

[Abdominal aortic thrombosis mimicking aortic coarctation in a newborn with heart failure]. / Trombose da aorta abdominal simulando coarctação da aorta em neonato com insuficiência cardíaca.

The case of a neonate in heart failure with the classical signs of coarctation of the aorta is described. Two dimension and Doppler echocardiography ruled out coarctation of the aorta and an abdominal ultrasonography detected a large thrombotic formation in the abdominal aorta, confirmed at necropsy.

Rupture of the aorta due to a malpositioned tracheal cannula in a 4-month-old baby.

Complications of tracheostomy are not rare and are even more common in children. They may be acute or chronic and sometimes may have serious consequences. We report here a rare situation in which a malpositioned tracheal cannula in a false tract from the neck to the mediastinum induced necrosis and rupture of the aorta and of the left carotid artery with fatal hemorrhage.

Translocation (1;5)(q32;q35) in CD30+ anaplastic large cell non-Hodgkin lymphona of childhood. A case report.

The authors studied cytogenetically a case of CD30+ anaplastic large cell non-Hodgkin lymphoma previously diagnosed as malignant histiocytosis and detected a translocation involving chromosomes 1 and 5, t(1;5)(q32:q35). After comparing their findings with those from reports in the literature, they comment about the importance of breakpoint q35 on chromosome 5 and point out the importance of associating morphologic, immunoperoxidase, and cytogenetic findings to confirm the diagnosis of this tumor.

Fístula aortoentérica primária / Primary aorto-enteric fistula

As fístulas aortoentéricas podem ser primárias ou secundárias. As últimas säo complicaçöes conhecidas de pacientes submetidos a cirurgias reconstrutivas da aorta. Enquanto estas säo relativamente comuns, as fístulas aortoentéricas primárias säo raras. Há cerca de 200 casos publicados na literatura mundial. Relata-se o caso de uma mulher de 71 anos com fístula aortoentérica primária que apresentava sangramento gatrointestinal de repetiçäo. O diagnóstico etiológico foi feito somente durante a cirurgi mediante o achado de um aneurisma aórtico comunicando-se com jejuno, apesar da exaustiva investigaçäo pré-operatória. Alguns aspectos clínicos e cirúrgicos dessa doença säo discutidos.

Schistosoma mansoni eggs in the placenta: an unusual finding

Schistosomiasis is a parasitic disease affecting millions of people in different parts of the world. It is estimated that only in Brazil 8 to 12 milion people are affected by Schistosoma mansoni, the species responsible for this disease in this country. Although its high prevalence, only rarely have S. mansoni eggs been found in the placenta. We report here a case in which the eggs were found in villous vessels during a routine examination of the placentas at our Institution. This region is not endemic for this disease and the patient did not know she had the disease