[Management of the first nonfebrile seizure in infants and children]. / Conduite à tenir devant une première crise épileptique du nourrisson et de l'enfant.

Seizures are the most common pediatric neurologic disorder. This article describes the guidelines of the French Pediatric Neurology Society, highlighting the importance of a thorough history and examination. Paroxysmal nonepileptic events should be excluded. The role of biological and neuroradiological investigations is discussed. An electroencephalographic recording and advice from a pediatric neurologist are suggested.

[Pseudoperiodic EEG heralding a cefepime-induced encephalopathy]. / Electroencéphalogramme pseudopériodique dans l'encéphalopathie due au céfépime.

Electroencephalography is a useful tool for the diagnosis and follow-up of toxic and metabolic encephalopathies. A pseudo-periodic pattern can indicate various brain pathologies and causes of mental confusion. Among these, cefepime encephalopathy should always be considered, particularly in cases of renal failure, because of the reversibility of the symptoms at drug withdrawal.

Identification and characterisation of a developmentally regulated mammalian gene that utilises -1 programmed ribosomal frameshifting.

Translational recoding of mRNA through a -1 ribosomal slippage mechanism has been observed in RNA viruses and retrotransposons of both eukaryotes and prokaryotes. Whilst this provides a potentially powerful mechanism of gene regulation, the utilization of -1 translational frameshifting in regulating mammalian gene expression has remained obscure. Here we report a mammalian gene, Edr, which provides the first example of -1 translational recoding in a eukaryotic cellular gene. In addition to bearing functional frameshift elements that mediate expression of distinct polypeptides, Edr bears both CCHC zinc-finger and putative aspartyl protease catalytic site retroviral-like motifs, indicative of a relic retroviral-like origin for Edr. These features, coupled with conservation of Edr as a single copy gene in mouse and man and striking spatio-temporal regulation of expression during embryogenesis, suggest that Edr plays a functionally important role in mammalian development.

Digitized mammography: a clinical trial of postmenopausal women randomly assigned to receive raloxifene, estrogen, or placebo.

BACKGROUND: High mammographic density is associated with increased breast cancer risk. Previous studies have shown that estrogens increase breast density on mammograms, but the effect on mammographic density of selective estrogen receptor modulators, such as raloxifene, is unknown. We assessed changes in mammographic density among women receiving placebo, raloxifene, or conjugated equine estrogens in an osteoporosis prevention trial. METHODS: In a 5-year multicenter, double-blind, randomized, placebo-controlled osteoporosis prevention trial, healthy postmenopausal women who had undergone hysterectomy less than 15 years before the study and had no history of breast cancer received placebo, raloxifene (at one of two doses), or conjugated estrogens (ERT). Women from English-speaking investigative sites who had baseline and 2-year craniocaudal mammograms with comparable positioning (n = 168) were eligible for this analysis. Changes in mammographic density were determined by digital scanning and computer-assisted segmentation of mammograms and were analyzed with the use of analysis of variance. All statistical tests were two-sided. RESULTS: Among the four treatment groups after 2 years on study, the mean breast density (craniocaudal view) was statistically significantly greater in the ERT group than it was in the other three groups (P<0.01 for all three comparisons). Within treatment groups, the mean breast density from baseline to 2 years decreased statistically significantly in women receiving the placebo or either the higher or lower raloxifene dose (P = 0.003, P = 0.002, and P<0.001, respectively) and showed a nonstatistically significant increase in women receiving ERT. CONCLUSIONS: In an osteoporosis prevention trial, raloxifene did not increase breast density after 2 years of treatment. Raloxifene administration should not interfere with, and could even enhance, mammographic detection of new breast cancers.

Cloning and developmental expression of Nsk2, a novel receptor tyrosine kinase implicated in skeletal myogenesis.

The protein superfamily of transmembrane receptor tyrosine kinases (RTKs) are essential components of intercellular signalling pathways necessary for normal cellular regulation. We report the cloning and developmental expression pattern of Nsk2, a novel, structurally distinct mammalian RTK characterised by a putative extracellular region bearing four immunoglobulin-like domains. The Nsk2 locus was mapped to the distal region of mouse chromosome 13 and was found to be expressed preferentially in skeletal muscle amongst adult mouse tissues. Moreover, increased steady-state levels of Nsk2 transcripts were apparent on terminal differentiation of committed skeletal myoblast cell lines in vitro and multiple isoforms of the Nsk2 RTK were identified in skeletal myotube cultures. RNA in situ hybridisation studies of mouse embryos confirmed skeletal myogenesis to be a major site of Nsk2 expression during normal embryogenesis, and identified other likely sites of Nsk2 function in ganglia of the developing peripheral nervous system and various embryonic epithelia, including those of kidney, lung and gut, during fetal development. Taken together, our data suggest normal functions for Nsk2 RTKs in distinctive aspects of skeletal muscle development, neurogenesis and mesenchymal-epithelial interactions during organ formation.

A juxtamembrane autophosphorylation site in the Eph family receptor tyrosine kinase, Sek, mediates high affinity interaction with p59fyn.

The large subfamily of receptor tyrosine kinases (RTKs) for which EPH is the prototype have likely roles in intercellular communication during normal mammalian development, but the biochemical signalling pathways utilised by this family are poorly characterised. We have now identified two in vitro autophosphorylation sites within the juxtamembrane domain of the Eph family member Sek, and a candidate binding protein for the activated Sek kinase. Specific antibodies defined Sek as a 130 kDa glycoprotein with protein kinase activity expressed in keratinocytes, whilst a bacterially expressed gst-Sek kinase domain fusion protein autophosphorylated exclusively on tyrosine residues, confirming that Sek encodes an authentic protein tyrosine kinase. Two dimensional phosphopeptide mapping and site-directed mutagenesis defined juxtamembrane residue Y602 as a major site of in vitro autophosphorylation in Sek, whilst Y596 was phosphorylated to a lower stoichiometry. Complimentary approaches of in vitro binding assays and BIAcore analysis revealed a high affinity association between the Y602 Sek autophosphorylation site and the cytoplasmic tyrosine kinase p59fyn, an interaction mediated through the SH2 domain of this intracellular signalling molecule. Moreover, these data identify the novel phosphotyrosyl motif pYEDP as mediating high affinity association with fyn-SH2, extending the previously defined consensus motif for this interaction. The extensive conservation of this fyn-binding motif within the juxtamembrane domain of Eph family RTKs suggests that signalling through fyn, or fyn-related, tyrosine kinases may be utilised by many members of this large subclass of transmembrane receptors.

Indicators of lifetime estrogen exposure: effect on breast cancer incidence and interaction with raloxifene therapy in the multiple outcomes of raloxifene evaluation study participants.

PURPOSE: To test the hypothesis that risk factors related to lifetime estrogen exposure predict breast cancer incidence and to test if any subgroups experience enhanced benefit from raloxifene. PATIENTS AND METHODS: Postmenopausal women with osteoporosis (N = 7,705), enrolled onto the Multiple Outcomes of Raloxifene Evaluation (MORE) trial, were randomly assigned to receive placebo, raloxifene 60 mg/d, or raloxifene 120 mg/d for 4 years. Breast cancer risk was analyzed by the following baseline characteristics indicative of estrogen exposure: previous hormone replacement therapy, prevalent vertebral fractures, family history of breast cancer, estradiol level, bone mineral density (BMD), body mass index, and age at menopause. Therapy-by-subgroup interactions were assessed using a logistic regression model. RESULTS: Overall, women with the highest one-third estradiol levels (> or = 12 pmol/L) had a 2.07-fold increased invasive breast cancer risk compared with women with lower levels. Raloxifene significantly reduced breast cancer risk in both the low- and high-estrogen subgroups for all risk factors examined (P <.05 for each comparison). The women with the highest BMD and those with a family history of breast cancer experienced a significantly greater therapy benefit with raloxifene, compared with the two thirds of patients with lower BMD or those without a family history, respectively; the subgroup-by-therapy interactions were significant (P =.005 and P =.015, respectively). CONCLUSION: The MORE trial confirms that increased lifetime estrogen exposure increases breast cancer risk. Raloxifene therapy reduces breast cancer risk in postmenopausal osteoporotic women regardless of lifetime estrogen exposure, but the reduction is greater in those with higher lifetime exposure to estrogen.

Flameless atomic absorption (FAA) and gas-liquid chromatographic studies in arsenic bioanalysis.

Procedures for assessment of arsenic in soft tissue by use of flameless atomic absorption (FAA) and gas-liquid chromatography (GLC), have been evolved, with special emphasis on the analytical distinction among inorganic, monomethyl-, and dimethylarsenic in several oxidation states. The chemical bases for such speciation reside in several properties of the arsenicals under consideration: (1) pentavalent inorganic arsenic, methylarsonic, and cacodylic acid are not extracted from tissue matter made strongly acid with hydrochloric acid, while the corresponding trivalent forms (as halides) are extracted; (2) chloroform extracts of samples treated under reducing conditions (HCl-KI) retain organoarsenicals when these extracts are re-extracted with water, but do not when aqueous solutions of oxidants are employed; (3) reduced cacodylate (dimethylarsinous acid) is not detected in the graphite furnace of an FAA unit under conditions selected, while cacodylate can be so detected. For GLC studies, monomethyl- and dimethylarsenic are simultaneously measured as the diethyldithiocarbamate complexes with an instrument equipped for electron-capture detection and containing a glass column packed with silanized 5% OV-17 on Anakrom A.S.

Effects of a phorbol ester on activation and immortalization of leukaemic B cells by Epstein-Barr virus.

Immortalization of chronic leukaemic B cells by Epstein-Barr virus (EBV) was investigated. Immortalization resistant and susceptible cell populations were defined by chronic lymphocytic leukaemia (CLL) and prolymphocytic leukaemia (PLL) cells respectively. These cell types could be distinguished by the effects of a phorbol ester on [3H]-thymidine incorporation by EBV-infected cells. A synergistic effect was observed in the non-immortalizing CLL samples, whereas this response was inhibited in the immortalizing PLL. Under the conditions used in the study, TPA did not cooperate with EBV to immortalize CLL cells.

Encoding and selective activation of "metabolic memories" in the rat.

Experiment 1 used Pavlovian conditioning procedures to show that rats formed distinct memorial representations of 2 (peanut oil and sucrose pellets) unconditioned stimuli (USs) that could be activated by 2 different conditioned stimuli (CSs). After training in Experiment 2, rats injected with the lipid antimetabolite Na-2-mercaptoacetate (MA) responded more to the CS for oil than to the CS for sucrose. This pattern was not shown by rats that received isotonic saline or systemic 2-deoxy-d-glucose (a glucose antimetabolite). By contrast, intracerebroventricular infusion of the glucose antimetabolite 5-thioglucose selectively promoted responding to the CS for sucrose (Experiment 4). Thus, lipoprivic and glucoprivic treatments selectively promoted the activation of the memories of fat and carbohydrate USs, respectively. In Experiment 3, the capacity of MA to augment responding to a CS for oil was abolished for rats that received subdiaphragmatic vagal deafferentation. This indicates that the capacity of lipoprivic signals to selectively activate the representations of fat USs may depend on vagal afferent fibers.

Does the integrated level of all plasma nutrients control daily food intake?

Studies on two different types of one-way crossed-intestines rats have shown that daily food intake is controlled by either endogenous gut signals or absorbed nutrients and their metabolic consequences, or both. If the amount of incoming ingested food is metered somewhere in the body, this could only occur in the gut or liver. The capacity of the liver to determine the amount of water-soluble nutrient absorbed was assessed by portacaval shunt and found to be inadequate. Infusion of nutrients directly into the bloodstream show that plasma nutrients provide part of the signal that inhibits daily food intake, but that endogenous gut signals must play some role. Insulin, an important hormone in the movement of plasma nutrients into cells, was found to stimulate food intake at low infusion doses. IV nutrients raise the level of plasma nutrients and lower daily food intake, while insulin, which inhibits the release of endogenous fuels and moves exogenous fuels into cells, lowers plasma nutrients and stimulates daily intake. Thus, the integrated level of all plasma nutrients may be a major controller of daily food intake.

Reliable method for cannulation of the third ventricle of the rat.

A method for chronic or acute cannulation of the third ventricle of the rat is described. This technique, in which successful cannulation is verified by the aspiration of a minute quantity of cerebrospinal fluid, is simple, reliable and produces minimal tissue distortion. The results of an experiment employing the technique and which compared the effects of direct intrahypothalamic and intraventricular administration of phentolamine on stimulus-bound feeding are briefly described.

Water intoxication death following hypothalamic lesions in the rat.

Rats received large, bilateral lesions of the ventromedial hypothalamus. Water or saline intakes, urine outputs and body temperatures were observed for up to 24 hr after surgery. Fifty percent of the operated animals drank excessively and died within 4-6 hr when permitted access to water. Urine outputs were low and symptoms of water intoxication were evident. When allowed access to saline, outputs rose and the number of animals which survived increased as the saline concentration increased. Body temperatures approached 40 degrees C during drinking, but did not differ from operated animals which refused to drink. It was concluded that the deposition of metallic ions strongly stimulates a hypothalamic drinking system which results in overhydration and water intoxication death.

Effect of intravenous nutrient infusions on food intake in rats.

To assess the effect of gut signals on food intake two types of nutrients were infused intravenously for 17.5 hours in 17 hour fed rats. In the first experiment a solution of 25% d-glucose and 4.25% amino acids (Travasol) was infused at levels of 26 and 52 kcal/day for two consecutive four-day periods. During infusion periods, food intake was reduced from saline baseline levels by 18.9 +/- 1.7 and 34.8 +/- 1.8 kcal/day, respectively. This represents an oral intake reduction of approximately 70% of the infused calories. In contrast, food intake was reduced 17.4 +/- 1.7 kcal/day below saline baseline levels when 40 kcal of Nutralipid were infused. The reduction in food intake was only 43% of the lipid calories infused. These results indicate that infusions of glucose and amino acids are more effective than infusion of fats in inhibiting daily food intake, that gut signals associated with absorption of fat provide important satiety signals and that removal of fat from the bloodstream has relatively little effect on daily food intake.

Insulin increases the daily food intake of diabetic rats on high and low fat diets.

The effects of insulin dose and diet composition on daily food intake were investigated by IV infusion of insulin in doses of 2 to 5 U/day into diabetic rats consuming either a high CHO or high fat diet. The daily food intake of the diabetic rats on both diets increased significantly over baseline levels (p < .01) at the low insulin doses and was maintained at these elevated levels through the 5 U/day dose. Insulin increased the rate of weight gain from Ig/day during baseline to 2 and 2.5 g/day in high CHO and high fat fed diabetics (p < .01). These results show that treatment of diabetic rats with continuous low doses of IV insulin results in a 40% increase in daily food intake regardless of the diet consumed and this increase is accompanied by an increase in rate of body weight gain. While the high fat fed diabetics were relatively hypoglycemic, these increases in intake are not the result of insulin-induced hypoglycemia, since blood glucose concentrations are significantly elevated when the increases occur at the lower insulin doses (p < .01). Thus, peripheralinsulin infused at physiological levels stimulates rather than inhibits daily food intake.

Hepatic portal and vena cava insulin infusion increase food intake in diabetic rats.

To test whether the route of insulin delivery has a major effect on the increase in daily food intake associated with chronic insulin treatment, insulin was continuously infused into either the vena cava (VC) or the hepatic portal (HP) vein of 23 diabetic Lewis rats. Increasing insulin doses in both the VC (2 to 6 U/day) and HP (1.5 to 3.5 U/day) groups significantly increased daily food intake (p < .05). Intake was higher in the VC group at 3 U/day but not at 2U/day. When insulin was delivered at a low fixed dose, daily food intake of both the VC and HP groups only increased after urinary glucose losses increased. The rate of weight gain increased significantly in the VC varied group (p < .05). Insulin administration also increased energy expenditure (p < .01). These results suggest that the extent of the increase in daily food intake and body weight that occurs with peripheral exogenous insulin administration is dependent on the route of infusion.

Insulin infusion stimulates daily food intake and body weight gain in diabetic rats.

Current theories state that physiological levels of insulin inhibit daily food intake and reduce body weight. To test whether insulin induces satiety, systematically increasing doses of insulin from 2.0 to 5.0 U/day were infused intravenously into streptozotocin-induced diabetic rats. Food intake increased significantly from 70.0 +/- 1.4 kcal/day during the saline baseline up to 102.2 +/- 1.9 kcal/day in the 3.5 U/day insulin infusion and then stabilized at 95.9 +/- 0.5 kcal/day for the remaining doses (p less than 0.05). Retained energy values (kcal of food intake minus kcal of urinary glucose loss) also increased from 69.9 +/- 1.4 kcal/day to stabilize at 95 kcal/day (p less than 0.001). Food intake and retained energy of normal controls remained unchanged at 75.4 +/- 1.6 kcal/day for the duration of the study. With elevated food intake and retained energy values after the 3.5 U/day insulin dose, the diabetic rats gained more weight than the normal controls (p less than 0.01). Contrary to expectations, increasing the amount of insulin infused through the physiological range results in a 40% increase in daily food intake and a rapid gain in body weight.

Adaptive calibration scheme for quantification of nutrients and byproducts in insect cell bioreactors by near-infrared spectroscopy.

Spectroscopic methods are gaining in popularity in biotechnology because of their ability to deliver rapid, noninvasive measurements of the concentrations of multiple chemical species. Such measurements are particularly necessary for the implementation of control schemes for cell culture bioreactors. One of the major challenges to the development of spectroscopic methods for bioreactor monitoring is the generation of accurate and robust calibration models, particularly because of the inherent variability of biological processes. We have evaluated several methods of building calibration models, including synthetic calibrations and medium spiking methods. The approach that consistently produced reliable models incorporated samples removed from a bioreactor that were subsequently altered so as to increase the sample variation. Several large volume samples were removed from a bioreactor at varying time points and divided into multiple aliquots to which were added random, known amounts of the analytes of interest. Near-infrared spectra of these samples were collected and used to build calibration models. Such models were used to quantify analyte concentrations from independent samples removed from a second bioreactor. Prediction errors for alanine, glucose, glutamine, and leucine were 1.4, 1.0, 1.1, and 0.31 mM, respectively. This adaptive calibration method produces models with less error and less bias than observed with other calibration methods. Somewhat more accurate measurements could be attained with calibrations consisting of a combination of synthetic samples and spiked medium samples, but with an increase in calibration development time.

The establishment and validation of the mobile immunization team concept at a clinic level.

Faced with unacceptably low and declining overall immunization compliance, as well as specific flu immunization compliance, the Executive Committee of a 32-person Air National Guard clinic asked the nursing service to devise a method of correcting these deficiencies as rapidly as possible and maintaining immunization compliance at a rate of 90% or better of the total patient population of approximately 1,100 full-time and Guard personnel for whom the clinic was responsible. The concept of a mobile immunization team was devised and validated over a 2-year interval. In order to successfully develop and implement this concept, command emphasis and a high level of cooperation from both clinic personnel and the Unit Commanders involved was requested and received. The make-up of the team, risk management, the timing and place of team visits, and record keeping were among the problems addressed and resolved. The mobile immunization team concept was a success, achieving excellent overall immunization compliance and outstanding compliance with the flu immunization program over a 2-year period.

[Indications of electroencephalogram in the newborn]. / Indications de l'électroencéphalogramme en période néonatale. Recommandations du groupe de neurophysiologie clinique de l'enfant.

The electroencephalogram (EEG), an easy-to-use and non invasive cerebral investigation, is a useful tool for diagnosis and early prognosis in newborn babies. In newborn full term babies manifesting abnormal clinical signs, EEG can point focal lesions or specific aetiology. EEG background activity and sleep organization have a high prognostic value. Tracings recorded over long period can detect seizures, with or without clinical manifestations, and differentiate them from paroxysmal non epileptic movements. The EEG should therefore be recorded at the beginning of the first symptoms, and if possible before any seizure treatment. When used as a neonatal prognostic tool, EEG background activity is classified as normal, abnormal (type A and type B discontinuous and hyperactive rapid tracing) or highly abnormal (inactive, paroxysmal, low voltage plus theta tracing). In such cases, the initial recording must be made between 12 and 48 h after birth, and then between 4 and 8 days of life. Severe EEG abnormalities before 12 h of life have no reliable prognostic value but may help in the choice of early neuroprotective treatment of acute cerebral hypoxia-ischemia. During presumed hypoxic-ischemic encephalopathy, unusual EEG patterns may indicate another diagnosis. In premature newborn babies (29-32 w GA) with neurological abnormalities, EEG use is the same as in term newborns. Without any neurological abnormal sign, EEG requirements depend on GA and the mother's or child's risk factors. Before 28 w GA, when looking for positive rolandic sharp waves (PRSW), EEG records are to be acquired systematically at D2-D3, D7-D8, 31-32 and 36 w GA. It is well known that numerous and persistent PRSW are related to periventricular leukomalacia (PVL) and indicate a bad prognosis. In babies born after 32 GA with clinically severe symptoms, an EEG should be performed before D7. Background activity, organization and maturation of the tracing are valuable diagnosis and prognosis indicators. These recommendations are designed (1) to get a maximum of precise informations from a limited number of tracings and (2) to standardize practices and thus facilitate comparisons and multicenter studies.