Visualization of two binding sites for the Escherichia coli UmuD'(2)C complex (DNA pol V) on RecA-ssDNA filaments.

The heterotrimeric UmuD'(2)C complex of Escherichia coli has recently been shown to possess intrinsic DNA polymerase activity (DNA pol V) that facilitates error-prone translesion DNA synthesis (SOS mutagenesis). When overexpressed in vivo, UmuD'(2)C also inhibits homologous recombination. In both activities, UmuD'(2)C interacts with RecA nucleoprotein filaments. To examine the biochemical and structural basis of these reactions, we have analyzed the ability of the UmuD'(2)C complex to bind to RecA-ssDNA filaments in vitro. As estimated by a gel retardation assay, binding saturates at a stoichiometry of approximately one complex per two RecA monomers. Visualized by cryo-electron microscopy under these conditions, UmuD'(2)C is seen to bind uniformly along the filaments, such that the complexes are completely submerged in the deep helical groove. This mode of binding would impede access to DNA in a RecA filament, thus explaining the ability of UmuD'(2)C to inhibit homologous recombination. At sub-saturating binding, the distribution of UmuD'(2)C complexes along RecA-ssDNA filaments was characterized by immuno-gold labelling with anti-UmuC antibodies. These data revealed preferential binding at filament ends (most likely, at one end). End-specific binding is consistent with genetic models whereby such binding positions the UmuD'(2)C complex (pol V) appropriately for its role in SOS mutagenesis.

Difference in early development of presumed monozygotic twins with Rett syndrome.

Normal early development has generally been insisted on as an essential criterion for the diagnosis of Rett syndrome. A new set of monozygotic female twins is reported. Twin 1 was considered to be abnormal from birth while delay was not suspected in twin 2 until she was about one year old. Some regression occurred during the second year in both twins, who are now clinically indistinguishable from each other at age 4 years. Other than a slight difference in head circumference at birth, no environmental factor which could account for the clinical difference has been identified.

Morphology of the oocyte-follicular connection in the mare.

The present study characterised the oocyte-follicular connection (i.e., oocyte fixation site) in Graafian follicles of the mare morphologically. Antral follicles were dissected in toto from ovaries obtained from oestrous, dioestrous and transitional mares after slaughter. The location of the cumulus oophorus complex in relation to the ovulation fossa, the width and density of the blood vessels surrounding the cumulus oophorus complex, the relative dimensions and histological aspects of the cumulus oophorus were investigated. For ultrastructural analysis of the junctional regions, cumulus-oocyte complexes were recovered in vivo by transvaginal ultrasound-guided follicle aspiration. The location of the oocyte fixation site was independent of mare, follicular size and stage of the oestrous cycle. In 82% of follicles, the oocytes were embedded in a broad based cell mount. The width and density of the blood vessels surrounding the oocyte fixation site were correlated to each other, but independent of follicular size and cyclic stage. The histological appearance of the cumulus oophorus varied, especially in respect to the compactness, and loosening of the cumulus cell population was observed in several medium-sized follicles from dioestrous mares. Loosening of the cumulus cell population was apparently associated with decreased interdigitation between adjacent corona radiata cells. It can be concluded that the fixation site of the equine cumulus oophorus complex represents a firm cellular anchorage between follicular wall and oocyte. Furthermore, the location of the cumulus oophorus complex in relation to the ovulation fossa and characteristics of the surrounding blood vessels is independent of follicular size and cyclic stage.

Seizures associated with calcifications and edema in neurocysticercosis.

We describe eight cases of pediatric patients whose neuroimages performed after seizures revealed abnormalities that were compatible with edema surrounding calcified lesions and which disappeared in subsequent examinations.

In vitro maintenance of Schistosoma japonicum and surgical transfer from donor to naïve recipient pigs.

An objective of this study was to find a culture medium and a temperature range suitable for in vitro maintenance of adult Schistosoma japonicum during surgical transplantation experiments. Adult S. japonicum were cultivated in four different media (NCTC 135, NCTC 109, RPMI 1640 and 0.85% physiological saline) supplemented with 10% heat-inactivated normal pig serum (hiNPS) at either 4 degrees C, 22-25 degrees C (room temperature) or 37 degrees C. Based on survival and morphologic evaluation, NCTC 135 at room temperature was found to be the best medium/temperature combination for maintenance of worms. An additional objective was to develop a method for transplanting adult S. japonicum from experimentally infected donor pigs to naïve recipient pigs. Six Landrace/Yorkshire crossbred pigs were used as donors to supply worms for two recipient pigs. Worms for transplantation were obtained by perfusion of the mesenteric veins of the donor pigs and maintained for a maximum of 3 h in NCTC 135 + 10% hiNPS at room temperature. A total of 148 and 132 worms were surgically transferred by way of an infusion tube into caecal veins of the two recipients. Six weeks after transplantation, 14% and 36% of the transferred worms were recovered by perfusion and subsequent manual inspection of the mesenteric veins of the two recipient pigs, respectively. The successful results suggest that surgical transfer of S. japonicum worms from donor to naïve recipient pigs may be useful for future studies on population genetics, dynamics and regulation in the pig/S. japonicum model.

Repeated transvaginal oocyte aspiration in unstimulated and FSH-treated mares.

Transvaginal ultrasound-guided follicular aspiration was conducted repeatedly in 5 cyclic mares. Three techniques were used and the aspirations were performed either > or = 23 d apart (A1, B1, C1) or 6 d apart (A2, B2,C2). During the A1 and A2 aspirations, the follicular cavity was flushed manually 8 to 10 times with flushing medium-filled (60 ml) syringes, while an electrical aspiration pump was used for the B1, B2, C1 and C2 aspirations. Prior to aspirations C1 and C2, the mares were treated daily with porcine FSH (100 mg, im) for 4 d. Aspiration was conducted 24 h after the last FSH injection. A total of 212 follicles with diameters varying between 5 and 23 mm was aspirated. The oocyte recovery rate was significantly higher (P = 0.004) from aspirations conducted with a > or = 23-d interval (35.8%) than from those aspirated 6 d after the previous aspiration (18.4%). Mode of evacuation and flushing (pump or syringe) and FSH treatment of the mare had no detectable effect on the oocyte recovery rate or on the cumulus dimensions of the aspirated oocytes. More than 80 % of the oocytes were at the germinal vesicle or diakinesis stage.

The influence of progesterone-induced proteins on glucose metabolism in early equine embryos.

The influence of different maternal plasma progesterone concentrations on embryonic glucose metabolism was studied. Uterine flushes were obtained after treating ovariectomized mares (n = 3) with 0 (control), 100 or 200 mg progesterone daily for 7 d. A group of progesterone-induced proteins (PIP) of Mr approximately 20,000 were identified in flushes from progesterone treatments by SDS-PAGE but were not observed in control flushes. Progesterone-induced proteins were removed from half the pooled flush in each treatment group by Sepharose blue CL-6B. In a 3 x 2 factorial (progesterone treatments, progesterone-induced proteins) experiment, 6 groups of Day 7 equine embryos (n = 6 per group) were incubated in culture media (MEM:DPBS; 1:3) containing radioactively-labeled glucose. Contributions of the Embden-Meyerhof pathway (EMP) and the pentose-phosphate pathway (PPP) to the total metabolism of glucose in early equine embryos were assessed separately. In the 200 mg progesterone treatment group, the presence of progesterone-induced proteins in the culture medium resulted in a 4-to 5-fold increase in the activities of the Embden-Meyerhof pathway and the pentose-phosphate pathway. These results lead to the following conclusions: 1)Addition of progesterone-induced uterine proteins from mares with high levels of circulating progesterone enhance the metabolic activities of the Embden-Meyerhof pathway and the pentose-phosphate pathway in Day 7.5 equine embryos in culture. 2)Uterine secretion of progesterone-induced proteins which is quantitatively and/or qualitatively adequate to modify embryonic glucose metabolism in vitro is dependent on a minimal concentration of maternal plasma progesterone.

Effect of two virus inactivation methods: electron beam irradiation and binary ethylenimine treatment on determination of reproductive hormones in equine plasma.

Ionizing irradiation and binary ethylenimine treatment have previously been shown to be effective for in-vitro inactivation of virus in biological material. In the present study the 2 methods were tested for possible effects on measurable concentrations of reproductive hormones in equine plasma (luteinizing hormone (LH), folliclestimulating hormone (FSH), progesterone (P4), and oestradiol-17 beta (E2)). The inactivation methods were electron beam irradiation with a dose from 11 to 44 kGy or treatment with binary ethylenimine (BEI) in concentrations of 1 and 5 mmol/L. Generally, there was a close correlation (r > 0.8, < 0.001) between pre- and post-treatment hormone levels. Thus, the different phases of the oestrous cycle could be distinguished on the basis of measured hormone concentrations of treated samples. However, both treatments significantly changed hormone concentrations of the plasma samples. For LH, FSH, and E2 the effect of irradiation and BEI treatment was depressive and dose-dependent. For P4 the effect of irradiation was also depressive and dose-dependent. However, the highest dose of BEI resulted in an increase of measured P4 concentration, which may be attributed to changes in the plasma matrix due to the treatment. Although the treatments affected measured hormone concentrations, the close correlation between pre-treatment and post-treatment measurements means that the diagnostic value will remain unchanged.

[Metachromatic leukodystrophy: report of 2 cases with histochemistry of nerves and muscles]. / Leucodistrofia metacromática: relato de dois casos com histoquímica de nervos e músculos.

Two cases of metachromatic leukodystrophy, of the late infantile form are reported. The patients were a girl and a boy of 2 years 10 months old, with initial normal development, but by the age of 18 months began with gait disturbances, difficulty to speak and developed progressive mental deterioration, with signs of long tract involvement, absence of deep tendon reflexes, spasticity, blindness, muscle atrophy and finished in a vegetative state. The diagnosis was made electromyography (signs of denervation), motor nerve conduction velocity (very decreased), assay of arylsulfatase A in the urine (absence of activity), sural nerve biopsy (demyelination and presence of metachromatic granules by the cresyl-violet and toluidine blue) and muscle biopsy (atrophy of type I fibers and presence of metachromatic material in the intramuscular nerve fibers). A quick revision about diagnostic methods, transmission, pathogenesis and variant forms is made.

[Neurocysticercosis in childhood. I. Clinical and laboratory diagnosis]. / Neurocisticercose na infância. I. Diagnóstico clínico e laboratorial.

Clinical and laboratory findings among 34 children (age range from 15 months to 13 years) with neurocysticercosis were reviewed. The main symptoms were: intracranial hypertension, 21 cases (62%); epilepsy, 20 cases (59%); hemiplegia, 4 cases (12%). Computed tomography (CT) in 33 children showed typical brain active cysts in 26 patients (79%) and calcifications in 2 (6%). The complement fixation reaction or the indirect immunofluorescent test for cysticercus antibody were positive in CSF in 77% (20 out of 26 patients) and serum in 78% (18 out of 23 patients). CSF pleocytosis was found in 57% of the cases (15/26 patients) with eosinophil cells in 27% (7/26 patients). The most efficient diagnostic test was CT scan of the head, and was confirmed immunologically by measurement of cysticercus antibody titers in CSF and serum.

[Neurocysticercosis in childhood. II. Computed tomography of 24 patients according to symptomatic and praziquantel treatment]. / Neurocisticercose na infância. II. Tomografia computadorizada de 24 pacientes em relação ao tratamento sintomático e com praziquantel.

We studied 24 children (15 months to 13 years old) which clinical, CSF and CT findings were compatible to the diagnosis of active neurocysticercosis. The patients were divided into three groups based on the type of treatment: Group I (10 patients) treated with analgesics and/or anticonvulsants; Group II (4 patients) treated with analgesics and or anticonvulsants and corticosteroids; Group III (10 patients) treated with analgesics and/or anticonvulsants, corticosteroids and praziquantel. The first patients CT scan were compared with the sequential CT scan findings (1 month to 5 years). The initial CT scan of 12 patients showed multiple active cysts, in 5 patients an isolated active cyst, in 3 patients partial calcified cysts, in 2 patients cerebral edema and in 2 patients were normal. The final results of the three groups of patients, as far as concern the normalization of CSF abnormalities or calcification of the cysts were the same, no matter the type of treatment applied to them. These results, although the small number of patients, showed that most of the children have good final results, with improvement of clinical symptoms and CT findings. We suggest that neurocysticercosis in children need multicenter study. So, a great number of patients can be followed and better definition can be established on the treatment of neurocysticercosis.

Cerebellar vermis hypoplasia - non progressive congenital ataxia: clinical and radiological findings in a pair of siblings.

We describe the clinical and radiological findings of a pair of siblings with cerebellar vermis hypoplasia and compare them with the literature. Both of them present pregnancies and deliveries uneventful and both presented some grade of hypotonia, ataxia, ocular motor abnormalities and mild motor delay and slurred speech. These siblings meet many of the criteria described in non-progressive congenital ataxia in which can occur familial cases with cerebellar atrophy, including vermis hypoplasia. As differential diagnosis we compare them with related syndromes and with Joubert's syndrome which main radiological finding on MRI is vermis hypoplasia associated with "molar tooth" appearance. The correct answer for these cases will only be possible by molecular genetics.

Bilateral perisylvian syndrome not related to malformations: report of two cases.

In this case report we present the neuroimaging findings and clinical features of two patients with a bilateral perisylvian syndrome not related to malformations, but probably to ischemic etiology. Evaluations including history, general and neurologic examinations, electroencephalograms, and imaging data were reviewed as recent literature about the subject.

[Rett syndrome: report of 7 cases]. / Síndrome de Rétt: relato de 7 casos.

The authors report the clinical and laboratory findings of seven girls with the diagnosis of Rett syndrome seen at the clinic from August 1978 to October 1988. The patient's age varied from two to ten years and they were followed up for 5 years and 9 months in average. The development milestone started to deteriorate between five and twenty-four months. Acquired microcephaly was present in six patients and three cases had hyperventilation. Generalized tonic-clonic seizures were the most frequent type of seizures (6 patients) beginning between one and seven years and effectively controlled with carbamazepine. The EEG's abnormal findings were spike discharges (5 patients) and paroxysmal high-amplitude theta activity (4 patients). The remaining laboratory tests were unremarkable. The pathogenesis of Rett syndrome has not been determined. The clinical evaluation is intended to contribute to the knowledge of that rather uncommon disease.

Sudden unexpected, unexplained death in epilepsy autopsied patients.

Sudden unexpected, unexplained death in epilepsy (SUDEP) has been reported to be responsible for 2 to 17% of all deaths in patients with epilepsy. This study was conducted to determine the circumstances of SUDEP and the autopsy findings in these patients. Fifty-three individuals whose cause of death was related to epilepsy were identified and in 30 cases relatives or friends were interviewed about the circumstances of death and other information which allowed to classify the patients as SUDEP or not. The death certificates were also reviewed. We found 20 cases of SUDEP. Most of them were found dead lying on the bed with no evidence of seizure event, and most of them had pulmonary and/or cerebral edema as the cause of death. The incidence and the risk of SUDEP can only be fully ascertained if all sudden deaths had postmortem examination. Consensus in certifying SUDEP cases would allow better accuracy in national mortality rate.

Epilepsy in children with cerebral palsy.

OBJECTIVE: To describe the prevalence and characteristics of epilepsy in patients with cerebral palsy in a tertiary center. METHODS: a total of 100 consecutive patients with cerebral palsy were retrospectively studied. Criteria for inclusion were follow-up period for at least 2 years. Types and incidence of epilepsy were correlated with the different forms of cerebral palsy. Other factors associated with epilepsy such as age of first seizure, neonatal seizures and family history of epilepsy were also analysed. RESULTS: follow-up ranged between 24 and 151 months (mean 57 months). The overall prevalence of epilepsy was 62%. Incidence of epilepsy was predominant in patients with hemiplegic and tetraplegic palsies: 70.6% and 66.1%, respectively. First seizure occurred during the first year of life in 74.2% of patients with epilepsy. Generalized and partial were the predominant types of epilepsy (61.3% and 27.4%, respectively). Thirty-three (53.2%) of 62 patients were seizure free for at least 1 year. Neonatal seizures and family history of epilepsy were associated with a higher incidence of epilepsy. CONCLUSIONS: epilepsy in cerebral palsy can be predicted if seizures occur in the first year of life, in neonatal period and if there is family history of epilepsy.

[Electron microscopy study in neurodegenerative diseases in childhood]. / Estudo por microscopia eletrônica em doenças neurodegerativas na infância.

Neurodegenerative diseases are a group of disorders in which there is storage of abnormal material in cells throughout the body due to an enzyme defect. The authors present the experience in the diagnosis of the neurodegenerative diseases in infancy by electron microscopical study of skin, conjunctival and rectum material of 89 patients and 2 necropsy cases. The age of the patients ranged from 49 days to 13 years with speak age of incidence in first year of life (n = 28). Fifty patients were female and 39 were male. The most frequent sites of biopsy were the skin and conjunctival. Of the total 89 patients, 15 had a definitive diagnosis (16.8%) including 4 cases of gangliosidosis, 3 cases of mucopolysaccharidosis, a case of Gaucher's disease, a case of Niemann-Pick disease, 3 cases of neuronal ceroid lipofuscinosis and 3 cases of storage disease which could not be specified. The authors studied all these patients within clinic and ultrastructural aspects and concluded that electron microscopy is an important method in diagnosis of storage diseases but with a low sensitivity as a single "screening" test for patients with progressive encephalopathy.

[Neuronal ceroid-lipofuscinosis. Report of 4 cases with study by rectal histochemistry, conjunctiva electron microscopy and necropsy]. / Lipofuscinose ceróide neuronal. Relato de quatro casos com estudo por histoquímica retal, microscopia eletrônica de conjuntiva e necrópsia.

The authors present the neuropathological and clinical findings of four cases of ceroid lipofuscinosis or Batten's disease. In two cases the autopsy findings showed brain atrophy and nerve cells packed with cytoplasmic granules compatible with lipofuscin. One case was diagnosed by histochemical techniques in frozen sections of rectal biopsy which showed myoenteric ganglion cells with cytoplasmic acid phosphatase positive material as well as numerous macrophages filled with lipofuscin in the mucosae. The fourth case diagnosed by electron microscopy of conjunctival biopsy which showed cytoplasmic inclusions of membranous, curvilinear and fingerprint types.

[Acute hepatic failure associated with valproic acid in children. Report of 3 cases]. / Insuficiência aguda hepática associada ao ácido valpróico na infância. Relato de três casos.

We report the cases of three epileptic children who developed hepatotoxicity induced by valproic acid. Two patients had developmental delay. Including the one who died, all patients were receiving polytherapy (carbamazepine in two and phenobarbital in one). The patients age ranged from 2 years and 8 months to 5 years and 1 month. The onset of hepatic complications occurred within 6 months of valproate therapy in two patients and 12 months in one. All patients developed the classical clinical signs of hepatotoxicity. Vomiting, edema and jaundice were the initial symptoms. Fever occurred in two patients. The serum levels of glutamic oxaloacetic transaminase were mildly elevated with a maximum of 194 IU. The bilirubin levels ranged from 5.5 to 19.8 mg%. Two patients recovered clinically and showed normalization of the laboratory abnormalities and one had fatal course. The hepatotoxicity must be considered as a side effect of valproic acid mainly in children under two years age, with polytherapy regimen and neurologic damage. The hepatic insufficiency can be reversible.

[Hypertensive encephalopathy associated to repetitive seizures: case report]. / Encefalopatia hipertensiva associada a estado de mal epiléptico de repetição: relato de caso.

Five years old, female, who started with tonic-clonic seizures on the right side of the body, with vomits and unconsciousness. The patient had been hospitalized for eight times in the last sixty days because of seizures. At physical exam, she had a severe arterial hypertension (270/140 mmHg). The computerized tomographic scan and magnetic resonance imaging revealed hypodense areas, mainly on the right parietal-temporal region, suggesting presence of edema. The angiography showed stenosis of the right renal artery, that was the cause of arterial hypertension. After the control of arterial hypertension by nephrectomy, the patient had a complete remission of the symptoms, as well as the images anomalies.