Two new missense mutations in the protein interaction ASH domain of OCRL1 identified in patients with Lowe syndrome.

The oculocerebrorenal syndrome of Lowe is a rare X-linked disease characterized by congenital cataracts, proximal renal tubulopathy, muscular hypotonia and mental impairment. This disease is caused by mutations in the OCRL gene encoding membrane bound inositol polyphosphate 5-phosphatase OCRL1. Here, we examined the OCRL gene of two Lowe syndrome patients and report two new missense mutations that affect the ASH domain involved in protein-protein interactions. Genomic DNA was extracted from peripheral blood of two non-related patients and their relatives. Exons and flanking intronic regions of OCRL were analyzed by direct sequencing. Several bioinformatics tools were used to assess the pathogenicity of the variants. The three-dimensional structure of wild-type and mutant ASH domains was modeled using the online server SWISS-MODEL. Clinical features suggesting the diagnosis of Lowe syndrome were observed in both patients. Genetic analysis revealed two novel missense variants, c.1907T>A (p.V636E) and c.1979A>C (p.H660P) in exon 18 of the OCRL gene confirming the clinical diagnosis in both cases. Variant c.1907T>A (p.V636E) was inherited from the patient's mother, while variant c.1979A>C (p.H660P) seems to have originated de novo. Analysis with bioinformatics tools indicated that both variants are pathogenic. Both amino acid changes affect the structure of the OCRL1 ASH domain. In conclusion, the identification of two novel missense mutations located in the OCRL1 ASH domain may shed more light on the functional importance of this domain. We suggest that p.V636E and p.H660P cause Lowe syndrome by disrupting the interaction of OCRL1 with other proteins or by impairing protein stability.

Novel missense mutation affecting the LIM-A domain of LMX1B in a family with Nail-Patella syndrome.

Nail-patella syndrome (NPS) is a rare autosomal dominant disease characterized by developmental defects of dorsal limb structures, the kidney, and the eye, that manifest as dysplastic nails, hypoplastic or absent patella, elbow dysplasia, iliac horns, glomerulopathy, and adult-onset glaucoma, respectively. This disorder is inherited in an autosomal dominant mode and is caused by heterozygous loss-of-function mutations in the LMX1B gene, which encodes the LIM homeodomain transcription factor LMX1B. In this study, we report the clinical findings of a Spanish family, from the Canary Islands, with three affected members who displayed varying phenotypes. DNA sequence analysis identified a novel heterozygous missense mutation in LMX1B, c.305A>G, p.(Y102C), that segregated with the disease. The tyrosine residue affected by the mutation is highly conserved in evolution, and is located in the LIM-A domain, next to one of the cysteine residues involved in zinc binding, suggesting that p.(Y102C) affects LMX1B function by disturbing its interactions with other proteins. Our results expand the mutation spectrum of LMX1B and provide insight into the molecular mechanisms of NPS pathology.

Pancreas Transplantation Delays the Progression of Morphological, Morphometric and Ultrastructural Changes in Testes of Alloxan-Induced Diabetic Rats.

Aim: The aim of this study was to assess the effects of pancreas transplantation on the progression of testicular lesions in diabetic rats. Methods: Diabetic rats were subjected to pancreas transplantation and sacrificed after 6, 14, 26 and 50 weeks of follow-up, using non-diabetic and untreated diabetic rats as controls. Results: Successful pancreas transplantation corrected all of the metabolic changes observed in diabetic rats, including low levels of testosterone. The testicular mass was decreased, and the relative weight of the testes was high in diabetic rats. The seminiferous tubules of diabetic rats showed progressive atrophy of the germinal epithelium, with cytoplasmic vacuolization, detachment of germ cells to the tubular lumen and the appearance of giant cells. Leydig cells were abnormally distributed, and hyperplasia of Sertoli cells was observed. Sperm were not detectable within the tubular lumen in late follow-up. The diameter, total area, lumen area, and germinal epithelium area of the seminiferous tubules were low, and tubular density was high in diabetic rats. Ultrastructural changes were also observed in these rats, compromising the cytoplasm, organelles and cellular nuclei of the germ, Sertoli, and Leydig cells. The most frequent changes consisted of accumulation of lipid droplets and electron-dense dark material in the cell cytoplasm, cellular degeneration and apoptosis. Similar to non-diabetic rats, pancreas-transplanted rats showed progressive testicular lesions, but they were much less severe and occurred much later than in the untreated diabetic controls. Conclusion: Diabetes causes morphological and ultrastructural changes in rat testes, but the progression of lesions can be significantly delayed by successful pancreas transplantation, which may have a positive impact on male infertility due to diabetes.

Torcicolo agudo como manifestação clínica isolada de histiocitose / Acute torticollis as a single clinical manifestation of histiocitosis

Menino, com 9 anos de idade, teve episódio agudo prolongado de torcicolo (laterocólis à esquerda), com duração de quinze dias. Radiografia de coluna evidenciou colapso da vértebra C5 e o diagnóstico final foi granuloma eosinofílico. O torcicolo pode ser agudo, recorrente ou prolongado. No adulto, a maioria dos casos de torcicolo recorrente é postural, mas na infância, torcicolos recorrentes ou prolongados tiveram possibilidade significativamente aumentada de haver uma causa, sendo asmais comuns lesões estruturais de coluna e lesões de neurônio motor primário.

A 9-year old boy presented an acute episode of torticollis (left deviation) that lasted for fifteen days. Conventional radiology of the cervical spine showed collapse of the C5 vertebra. His conclusive diagnosis was thatof eosinophilic granuloma. Torticollis can be acute recurrent or prolonged. In adulthood, most cases of recurrent torticollis are caused by postural problems; however, in childhood, recurrent or prolonged torticollis is much more likely to have a cause, and the most common are spine structural lesions or lesions in the primary motor neuron.

Long term evaluation of morphometric and ultrastructural changes of testes of alloxan-induced diabetic rats.

PURPOSE: To evaluate in a long term the morphometric and ultrastructural changes in seminiferous tubules (ST) of normal and diabetic rats, and to correlate any changes with animal age and diabetes duration. METHODS: Sixty male Wistar rats, three months-old, were randomly divided into two groups: 30 non-diabetic controls (N) and 30 alloxan untreated diabetic (D). After one, six and 12 months of follow-up or diabetes induction rats were sacrificed and the testes examined. Morphometric measures of the ST were performed by digital imaging analysis. ST ultrastructure was analyzed by transmission electron microscopy. RESULTS: Sustained hyperglycemic state was observed in all diabetic rats throughout the study. Serum testosterone was also significantly decreased in these animals. The diameter, total area, epithelium area and epithelium thickness of ST were lower and tubular density was higher in diabetic animals. Diabetic rats also showed ultrastructural changes compromising the whole testis including germ-, Sertoli-, and Leydig cells, and also the mithocondria and cellular nuclei. Most frequent of these consisted of vacuolization and/or accumulation of lipid droplets and electron dense dark material in cell cytoplasm and/or in membranes, cellular degeneration, and apoptosis. Non-diabetic control rats also showed testicular lesions that resemble to the diabetic lesions, although much less severe and with later onset in life of these animals. CONCLUSION: Histopathological changes observed in testes of normal and diabetic rats are closely related to the animal age and/or duration of the hyperglycemic state, being progressively more severe in animals sacrificed belatedly. These changes may play an important role in male infertility observed in diabetic subjects.

Long term evaluation of morphometric and ultrastructural changes of testes of alloxan-induced diabetic rats

PURPOSE: To evaluate in a long term the morphometric and ultrastructural changes in seminiferous tubules (ST) of normal and diabetic rats, and to correlate any changes with animal age and diabetes duration. METHODS: Sixty male Wistar rats, three months-old, were randomly divided into two groups: 30 non-diabetic controls (N) and 30 alloxan untreated diabetic (D). After one, six and 12 months of follow-up or diabetes induction rats were sacrificed and the testes examined. Morphometric measures of the ST were performed by digital imaging analysis. ST ultrastructure was analyzed by transmission electron microscopy. RESULTS: Sustained hyperglycemic state was observed in all diabetic rats throughout the study. Serum testosterone was also significantly decreased in these animals. The diameter, total area, epithelium area and epithelium thickness of ST were lower and tubular density was higher in diabetic animals. Diabetic rats also showed ultrastructural changes compromising the whole testis including germ-, Sertoli-, and Leydig cells, and also the mithocondria and cellular nuclei. Most frequent of these consisted of vacuolization and/or accumulation of lipid droplets and electron dense dark material in cell cytoplasm and/or in membranes, cellular degeneration, and apoptosis. Non-diabetic control rats also showed testicular lesions that resemble to the diabetic lesions, although much less severe and with later onset in life of these animals. CONCLUSION: Histopathological changes observed in testes of normal and diabetic rats are closely related to the animal age and/or duration of the hyperglycemic state, being progressively more severe in animals sacrificed belatedly. These changes may play an important role in male infertility observed in diabetic subjects.

Alterações testiculares no diabetes aloxânico no rato. Estudo morfométrico e ultraestrutural dos túbulos seminíferos / Testicular alterations in diabetic rats alloxan: ultrastructural and morphometric study of seminiferous tubules

Este estudo foi realizado com o objetivo de avaliar, no longo prazo, os parâmetros morfométricos e ultraestruturais dos tubulos seminíferos dos testículos de ratos diabéticos aloxânicos. 60 ratos Wistar, machos, pesando de 250 a 280 gramas foram divididos em 2 grupos experimentais: N ou grupo normal, constituído de 30 animais sadios, nãodaibéticos; D ou grupo diabético, constituído de 30 animais diabéticos sem qualquer tratamento da doença. Cada grupo experimental foi posteriormente dividido, por sorteio, em 3 subgrupos de ratos, com 10 animais cada um, para serem avaliados e sacrificados com 1, 6 e 12 meses de seguimento. O diabetes experimental foi induzido pela administração endovenosa de aloxana 2%, na dose de 42 mg/kg de peso corporal. Somente animais com sinais clínicos evidentes de diabetes grave e com glicemia >250 mg/dl e glicose urinária > 3000 mg/dl foram utilizados neste experimento. Com 1, 6 e 12 meses de seguimento o peso corporal, a ingestão hídrica, a ingestão alimentar, a diurese e os níveis de glicose sanguínea foram avaliados em todos os animais. Com 1 e 6 meses de seguimento também foram analisados os níveis de insulina plasmática, hemoglobina glicosilada (HbA1C) e de testosterona sérica Após os sacrifício dos animais, ambos os testículos foram removidos, sendo o testículo esquerdo preparado para a microscopia ótica e o direito para a microscopia eletrônica. As medidas morfométricas à M.O. e M.E. foram realizadas por análise digital de imagens utilizando o software Quin Lite 2.5 - LEICA . A ultraestrutura dos túbulos seminíferos foi analisada ao microscópio de transmissão PHILIPS, pela análise descritiva comparativa.(...)

This study was conducted to evaluate, in the long term, ultrastructural and morphometric parameters of seminiferous tubules of the testis of diabetic rats alloxan. 60 male Wistar rats weighing 250-280 grams were divided into two groups: group N or normal, consisting of 30 healthy animals, non-daibetics; D or diabetic group, consisting of 30 diabetic animals without in treatment. Each experimental group was then divided in three subgroups of rats with 10 animals each to be evaluated and sacrificed at 1, 6 and 12 months of follow-up. Experimental diabetes was induced by intravenous administration of alloxan 2% at a dose of 42 mg / kg body weight. Only animals with obvious signs of severe diabetes and blood glucose> 250 mg / dl and urine glucose> 3000 mg / dl were used in this experiment. At 1, 6 and 12 months of follow-up body weight, water intake, food intake, diuresis and blood glucose levels were evaluated in all animals. At 1 and 6 months of follow-up were also analyzed the levels of plasma insulin, glycosylated hemoglobin (HbA1C) and serum testosterone after the animal sacrifice, both testis were removed, the left was prepared for light microscopy and the right to the electron microscopy. Morphometric measures at M.Õ. and ME were performed by using digital image analysis software Quin Lite 2.5 - LEICA. The ultrastructure of the seminiferous tubules was analyzed by transmission microscope PHILIPS, by comparative descriptive analysis.(...)

Estudo da excreção urinária de cálcio, potássio e sódio com o emprego de citrato de potássio na hipercalciúria idiopática na criança / Study of urinary excretion of calcium, potassium and sodium using potassium citrate in children with idiopathic hypercalciuria

OBJETIVO: Estudar as relações entre a excreção urinária de cálcio, sódio e potássio e a associação sódio/potássio urinários em crianças com hipercalciúria idiopática em dieta habitual, antes e depois da administração de citrato de potássio na dose de 1mEq/kg/dia. MÉTODOS: Foram estudadas prospectivamente 26 crianças: 19 (73 por cento) meninos e sete (27 por cento) meninas com idade entre dois e 13 anos, portadores de hipercalciúria idiopática recém-diagnosticada por dosagem de cálcio em urina de 24 horas >4mg/kg/dia. O citrato de potássio foi administrado na dose de 1mEq/kg/dia. Foram realizadas dosagens séricas e em urina de 24 horas de cálcio (Ca), potássio(K), sódio (Na) e creatinina (Cr), antes e 15 dias depois da administração diária do citrato de potássio. Para comparar os resultados de cálcio/creatinina (Ca/Cr), potássio/creatinina (K/Cr) e sódio/potássio (Na/K) urinários nos dois momentos, aplicou-se o teste não-paramétrico de Wilcoxon. Para a análise das associações entre Ca/Cr e K/Cr e entre Ca/Cr e Na/Cr foi utilizado o coeficiente de correlação de Pearson. Considerou-se significante p<0,05. RESULTADOS: Após o uso de citrato de potássio, ocorreu significativa redução da calciúria e da relação Na/K urinários, bem como elevação na caliúria. Não houve modificação da excreção urinária de sódio. CONCLUSÕES: Em dieta habitual, o citrato de potássio eleva a caliúria e diminui a calciúria em criança hipercalciúricas, sendo um eficaz recurso terapêutico.

OBJECTIVE: Evaluate the relationships among the urinary excretion of calcium (UCa), potassium (UK), sodium (UNa) and the ratio between UNa/UK in children with idiopathic hypercalciuria and a regular diet, before and after 1mEq/kg/day potassium citrate administration. METHODS: 26 children with idiopathic hypercalciuria (UCa>4mg/kg/day) were prospectively studied: 19 (73 percent) boys and seven (27 percent) girls between two and 13 years old. Potassium citrate was administered: 1mEq/Kg/day twice a day for 15 days. Blood and 24-hour urinary determinations of calcium, potassium, sodium and creatinine were done in two periods: before and after the 15-day administration of potassium citrate. The following urinary ratios were analyzed before and after potassium citrate use by Wilcoxon test: calcium/creatinine (UCa/UCr), potassium/creatinine (UK/UCr) and sodium/creatinine (UNa/UCr). The association between UCa/UCr, UK/UCr and Ca/Cr, UCa/UCr and UNa/UK were analyzed by Pearson's correlation. Significance was considered for p<0.05. RESULTS: After potassium citrate use, there were significant reductions of UCa and UNa/UK ratios, as well as a significant increase of UK. The UNa did not change. CONCLUSIONS: Children with idiopathic hypercalciuria and regular diet treated with daily potassium citrate increased their potassium urinary excretion and decreased their calciuria.

Vasculite necrosante na glomerulonefrite difusa aguda pós-infecciosa / Necrotizing vasculitis in acute postinfectious glomerulonephritis

Os autores relatam dois casos de glomerulonefrite difusa aguda pós-infecciosa com evolução clinicomorfológica incomum. As biópsias renais mostraram alterações características de glomerulonefrite difusa aguda associada à extensa necrose fibrinóide e infiltrado inflamatório leucocitário na parede de arteríolas e artérias interlobulares. Foram também observadas crescentes. Ambos os pacientes cursaram com insuficiência renal aguda severa, sendo que um dos pacientes recuperou a função renal e outro evoluiu para insuficiência renal crônica e óbito.

Insuficiência renal crônica na criança: aspectos clínicos, achados laboratorial e evolução / Chronic renal failure in children: clinical features, laboratory findings and outcome

Objetivo :Descrever os aspectos clínicos, laboratoriais e a evolução de crianças com insuficiência renal crônica. Métodos :Estudo descritivo de crianças com insuficiência renal crônica no período entre 1972 e 2000. Resultados :Foram estudados 45 pacientes, dos quais 58,7(por cento) eram do sexo masculino e 53,4(por cento) com idade superior a 7 anos. Observamos alteração do peso e estatura em 37,7(por cento) e 42,2(por cento) dos pacientes, respectivamente. As alterações laboratoriais mais encontradas foram acidose metabólica em 93,8(por cento) dos casos, insuficiência renal moderada a grave em 66,6(por cento) dos casos e IRC terminal em 28,8(por cento) dos casos. As causas principais foram malformação do trato urinário em 48,8(por cento) dos casos e glomerulopatias em 40(por cento) dos casos. Crianças em tratamento conservador ou dialítico apresentaram crescimento insatisfatório. Óbito ocorreu em 6 crianças (15,4(por cento)). Conclusão:O estudo constou do predomínio de crianças do sexo masculino, com idade acima de 7 anos, com anormalidades do trato urinário e alta percentagem de insuficiência renal crônica terminal, cujas alterações clínicas e laboratoriais graves podem ter contribuído para peso e estatura baixos.

Infecção do trato urinário na criança - estudo retrospectivo / Urinary infection in child-retrospective study

A infecção urinária é patologia freqüente em crianças e sua importância está na existência do risco de complicações em pacientes com anomalias congênitas e refluxo vesico-ureteral com dilatação, que, associados à repetição da infecção urinária, podem levar ao desenvolvimento de cicatrizes renais, com conseqüente presença de hipertensão arterial e/ou insuficiência renal. Objetivo: Relatar casuística retrospectiva de 155 crianças com infecção urinária. Métodos: Os dados foram obtidos por meio do preenchimento de protocolos de crianças de 1 a 122 meses de idade, média de 45,2 meses, atendidas durante o período de janeiro de 1989 a dezembro de 2000. Resultados: Observamos que 71 por cento foram do sexo feminino e 41,9 por cento com idade inferior a 2 anos. Cento e doze pacientes (72,2 por cento) apresentaram febre, 14,2 por cento disúria e 13,5 por cento dor abdominal. O agente etiológico foi a E. coli em 73,5 por cento dos pacientes. Dilatação da pelvis renal (20,4 por cento) e refluxo vésico-ureteral (38,1 por cento) foram as alterações mais freqüentes observadas nos estudos por imagem. Sessenta e um pacientes (39,4 por cento) apresentaram o trato urinário normal. Sessenta e oito pacientes (43,9 por cento) apresentaram infecção de repetição e o agente foi a E. coli em 47,5 por cento e Proteus em 22,8 por cento. Das 68 crianças com infecção de repetição, 42 (61,7 por cento) apresentaram anormalidades do trato urinário. Conclusão: A infecção urinária foi observada em maior porcentagem em crianças do sexo feminino; na faixa etária até 2 anos; tendo como principal quadro clínico febre, constatando-se anormalidades do trato urinário associadas à presença de recidivas da infecção.