Efficient gene expression from integration-deficient lentiviral vectors in the spinal cord.

Gene transfer to spinal cord cells may be crucial for therapy in spinal muscular atrophy, amyotrophic lateral sclerosis and spinal cord injury. Lentiviral vectors are efficient for transduction of a variety of cells, but like all integrating vectors they pose a risk of insertional mutagenesis. Integration-deficient lentiviral vectors (IDLVs) remain episomal but retain the transduction efficiency of standard integrating lentiviral vectors, particularly when the episomes are not diluted out through repeated cell division. We have now applied IDLVs for transduction of spinal cord in vitro, in explants and in vivo. Our results demonstrate similar efficiency of eGFP expression from integrating lentiviral vectors and IDLVs in most cell types analyzed, including motor neurons, interneurons, dorsal root ganglia (DRG) neurons and astroglia. IDLV-mediated expression of pro-glial-cell-derived neurotrophic factor (Gdnf) rescues motor neuron cultures from death caused by removal of exogenous trophic support. IDLVs also mediate efficient RNA interference in DRG neuron cultures. After intraparenchymal injection in the rat and mouse cervical and lumbar regions in vivo, transduction is mainly neuronal, with both motor neurons and interneurons being efficiently targeted. These results suggest that IDLVs could be efficient and safer tools for spinal cord transduction in future therapeutic strategies.

Pronóstico a largo plazo de la epilepsia ausencia juvenil / Long term prognosis of juvenile absence epilepsy

Introducción: La epilepsia ausencia juvenil (EAJ) es un tipo de epilepsia generalizada idiopática que se caracteriza por la presencia de crisis de ausencia (CA) que comienzan en la adolescencia, con un EEG típico de punta-onda generalizada, y que puede acompañarse de mioclonías y crisis tónico-clónicas generalizadas (CTCG). El pronóstico a largo plazo es incierto. Material y métodos: Seleccionamos de manera retrospectiva todos los pacientes que cumplían los criterios diagnósticos de EAJ de la ILAE 1989, analizamos las variables clínicas, el tratamiento farmacológico, el estar libre de crisis y la posibilidad de retirar el tratamiento. Resultados: Encontramos 21 pacientes, 17 mujeres y 4 varones, el 86% presentó también CTCG y el 14% mioclonías. La edad de inicio de las CA fue de 17 años (rango: 10-44). Cuatro pacientes comenzaron con CA en la edad adulta. El seguimiento medio fue de 25 años (intervalo: 10-43). El 90% recibió tratamiento con valproato y el 62% requirió politerapia. El 43% de los pacientes están actualmente libres de crisis, aunque todos ellos en tratamiento farmacológico. Todos los intentos de retirar la medicación fracasaron, pese a largos períodos sin crisis.Conclusiones: Menos de la mitad de los pacientes con EAJ están libres de crisis. El tratamiento antiepiléptico es necesario durante toda la vida a pesar de largos períodos de remisiones (AU)

Introduction: Juvenile absence epilepsy (JAE) is a generalized form of epilepsy, characterizedby absence seizures (AS) initiated in adolescence, with a typical EEG showing generalized spikewavedischarges. Apart from absences, other seizure types may be observed such as myocloniaand generalized tonic-clonic seizures (GTCS). Its long-term prognosis is uncertain Material and methods: We retrospectively selected all patients who met the 1989 ILAE diagnostic criteria for JAE. We analysed clinical variables, pharmacological treatment, and seizure remission with medical treatment and seizure relapse after stopping medical treatment. Results: We identified 21 patients, 17 women and 4 men, 86% of whom had suffered GTCS and 14% myoclonias. Mean age at AS onset was 17 years old (range 10-44), 4 patients debuted with AS in adulthood. Mean follow up duration was 25 years (range 14-43). Ninety per cent of the patients were treated with valproate and 62% needed polytherapy. Currently 43% have achieved seizure freedom under medical treatment. All attempts to stop treatment failed, in some cases after long periods of seizure remission. Conclusions: Less than fifty per cent of patients with JAE achieve remission, antiepileptictreatment is mandatory during all life, despite having long periods of remission (AU)

Long term prognosis of juvenile absence epilepsy.

INTRODUCTION: Juvenile absence epilepsy (JAE) is a generalized form of epilepsy, characterized by absence seizures (AS) initiated in adolescence, with a typical EEG showing generalized spike-wave discharges. Apart from absences, other seizure types may be observed such as myoclonia and generalized tonic-clonic seizures (GTCS). Its long-term prognosis is uncertain. MATERIAL AND METHODS: We retrospectively selected all patients who met the 1989 ILAE diagnostic criteria for JAE. We analysed clinical variables, pharmacological treatment, and seizure remission with medical treatment and seizure relapse after stopping medical treatment. RESULTS: We identified 21 patients, 17 women and 4 men, 86% of whom had suffered GTCS and 14% myoclonias. Mean age at AS onset was 17 years old (range 10-44), 4 patients debuted with AS in adulthood. Mean follow up duration was 25 years (range 10-43). Ninety per cent of the patients were treated with valproate and 62% needed polytherapy. Currently 43% have achieved seizure freedom under medical treatment. All attempts to stop treatment failed, in some cases after long periods of seizure remission. CONCLUSIONS: Less than fifty per cent of patients with JAE achieve remission, antiepileptic treatment is mandatory during all life, despite having long periods of remission.

Tumor necrosis factor alpha and interferon gamma cooperatively induce oxidative stress and motoneuron death in rat spinal cord embryonic explants.

The accumulation of reactive microglia in the degenerating areas of amyotrophic lateral sclerosis (ALS) tissue is a key cellular event creating a chronic inflammatory environment that results in motoneuron death. We have developed a new culture system that consists in rat spinal cord embryonic explants in which motoneurons migrate outside the explant, growing as a monolayer in the presence of glial cells. The proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) have been proposed to be involved in ALS-linked microglial activation. In our explants, the combined exposure to these cytokines resulted in an increased expression of the pro-oxidative enzymes inducible nitric oxide synthase (iNOS), the catalytic subunit of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, gp91(phox) and cyclooxygenase-2 (COX-2), as compared to each cytokine alone. This effect was related to their cooperation in the activation of the transcription factor nuclear factor kappa B (NF-kappaB). TNF-alpha and IFN-gamma also cooperated to promote protein oxidation and nitration, thus increasing the percentage of motoneurons immunoreactive for nitrotyrosine. Apoptotic motoneuron death, measured through annexin V-Cy3 and active caspase-3 immunoreactivities, was also found cooperatively induced by TNF-alpha and IFN-gamma. Interestingly, these cytokines did not affect the viability of purified spinal cord motoneurons in the absence of glial cells. It is proposed that the proinflammatory cytokines TNF-alpha and IFN-gamma have cooperative/complementary roles in inflammation-induced motoneuron death.

Vascular endothelial growth factor protects motoneurons from serum deprivation-induced cell death through phosphatidylinositol 3-kinase-mediated p38 mitogen-activated protein kinase inhibition.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the selective degeneration and death of motoneurons in the spinal cord, brainstem and motor cortex which causes progressive muscle weakness and paralysis. Although the molecular mechanisms causing the disease remain unknown, excitotoxicity and loss of trophic support have been proposed as causes of degeneration. The present study was designed to elucidate the mechanisms of motoneuron death induced by serum deprivation and the potential neuroprotective effects of vascular endothelial growth factor (VEGF) in dissociated and organotypic rat spinal cord cultures. Serum withdrawal induced apoptotic cell death in dissociated spinal cord cultures, which was prevented in the presence of VEGF. In organotypic spinal cord cultures, low serum-induced motoneuron death was mediated by the stress-related kinase p38 mitogen-activated protein kinase (p38MAPK), as it was reversed by the p38MAPK inhibitor SB203580. In these cultures, exposure to VEGF blocked p38MAPK phosphorylation and prevented the demise of motoneurons. These effects of VEGF were mediated through the phosphatidylinositol 3-kinase/Akt (PI3-K/Akt) signal transduction pathway, as they were blocked in the presence of the PI3-K inhibitor LY294002. In addition, serum deprivation induced down-regulation of the anti-apoptotic protein Bcl-2 and this effect was prevented both by SB203580 and by VEGF via the PI3-K/Akt pathway. Therefore, Bcl-2 could also play an important role in the neuroprotection induced by VEGF in spinal cord cultures. Together, these findings indicate that VEGF prevents motoneuron death induced by serum deprivation blocking the activity of p38MAPK via the PI3-K/Akt signaling pathway.

[Fetal macrosomia: management, obstetrical and neonatal results. Multicenter case-control study in 15 maternity hospitals in Paris and the Ile de France area]. / Macrosomie foetale: pratiques, conséquences obstétricales et néonatales. Enquête multicentrique cas-témoins menée dans 15 maternités de Paris et d'Ile de France.

OBJECTIVE: To study the perinatal management of fetal macrosomia (FM) and the obstetrical and neonatal results related to FM in the Ile de France area. MATERIALS AND METHODS: Case-control study from the fifteenth of July to the fifteenth of September 1999 in fifteen maternity in Paris and the Ile de France area. All singletons, without malformation, weighing more than 4,000 grams, born after 37 weeks of pregnancy during the study were included. The control group had the same inclusion and exclusion criteria (except the birth-weight) and was defined by the next delivery of same parity. RESULTS: 384 FM and 384 controls have been included. Usual risk factors of macrosomia have been found. The screening of gestational diabetes was realised in 56.8% and FM was suspected before delivery in 59.3% in the FM group. In cases of FM, the midwife was alone at the time of delivery in 53.4% of spontaneous vaginal delivery. FM was associated with a longer labour and a more frequent use of oxytocin. There was six times more severe perineal tears (1.7 vs 0.3%; p = 0.05) for women with FM whereas the rate of haemorrhage at delivery was the same in both groups. Cesarean section' rate before and during labor was higher in the FM group whereas instrumental extraction was not different. In this study, FM was not associated with an excess of fetal morbidity (injury, Apgar score, pH cord) even if we found ten times more shoulder dystocia. CONCLUSION: Complications related to FM were mainly maternal in this study. Some recommendations accounting fetal macrosomia were not widely adopted as screening of gestational diabetes or necessity to have a whole obstetric team at the time of delivery.

Long-lasting aberrant tubulovesicular membrane inclusions accumulate in developing motoneurons after a sublethal excitotoxic insult: a possible model for neuronal pathology in neurodegenerative disease.

We have previously shown that chronic treatment of chick embryos [from embryonic day 5 (E5) to E9] with NMDA rescues spinal cord motoneurons (MNs) from programmed cell death. In this situation, MNs exhibit a reduced vulnerability to acute excitotoxic lesions and downregulate NMDA and AMPA-kainate receptors. Here, we report that this treatment results in long-lasting sublethal structural changes in MNs. In Nissl-stained sections from the spinal cord of NMDA-treated embryos, MNs display an area adjacent to an eccentrically positioned nucleus in which basophilia is excluded. Ultrastructurally, MNs accumulate tubulovesicular structures surrounded by Golgi stacks. Thiamine pyrophosphatase but not acid phosphatase was detected inside the tubulovesicular structures, which are resistant to disruption by brefeldin A or monensin. Immunocytochemistry reveals changes in the content and distribution of calcitonin gene-related peptide, the KDEL receptor, the early endosomal marker EEA1, and the recycling endosome marker Rab11, indicating that a dysfunction in membrane trafficking and protein sorting occurs in these MNs. FM1-43, a marker of the endocytic pathway, strongly accumulates in MNs from isolated spinal cords after chronic NMDA treatment. Changes in the distribution of cystatin C and presenilin-1 and an accumulation of amyloid precursor protein and beta-amyloid product were also observed in NMDA-treated MNs. None of these alterations involve an interruption of MN-target (muscle) connections, as detected by the retrograde tracing of MNs with cholera toxin B subunit. These results demonstrate that chronic NMDA treatment induces severe changes in the motoneuronal endomembrane system that may be related to some neuropathological alterations described in human MN disease.

Screening with a uterine Doppler in low risk pregnant women followed by low dose aspirin in women with abnormal results: a multicenter randomised controlled trial.

OBJECTIVES: To assess whether systematic screening with an uterine artery Doppler in low risk pregnant women followed by the prescription of low dose aspirin in cases with abnormal results reduced the incidence of intrauterine growth restriction and pre-eclampsia. DESIGN: A multicentre randomised trial. POPULATION: 3,317 low risk pregnant women. In the Doppler group, the uterine artery Doppler was performed between 20 and 24 weeks. Women with abnormal results received 100 mg of aspirin daily until the 35th week. MAIN OUTCOME MEASURES: Intrauterine growth restriction was defined as birthweight below the tenth and the third centile according to gestational age. Pre-eclampsia was defined as hypertension associated with proteinuria > 0.5g/L. RESULTS: Intrauterine growth restriction, whether defined by the third or tenth centile, did not differ significantly between the two groups (RR = 1.22 [0.73 - 2.04] and 1.18 [0.93 - 1.51] respectively). Screening with uterine artery Doppler did not affect birthweight or any of the criteria of perinatal morbidity. There was no effect on the incidence of pre-eclampsia (RR = 1.99 [0.97 - 4.09]) or hypertensive disorders. These results were the same for nulliparae and multiparae. CONCLUSIONS: There is no justification for screening with uterine artery Doppler in a low risk population, even if abnormal results are followed by aspirin treatment and increased prenatal surveillance. Future studies must assess predictive tests that can be performed early in pregnancy and can identify populations at very high risk of pre-eclampsia and intrauterine growth restriction. Only when all of these conditions are fulfilled, aspirin or other treatments may prove its efficacy.

Sudden unexpected death in infants under 3 months of age and vaccination status- -a case-control study.

AIMS: To determine whether DTPP+Hib vaccination (diphtheria, tetanus, pertussis, poliomyelitis +/- haemophilus) increased the risk of sudden unexpected death (SUD) in children under 3 months of age. METHODS: We conducted a multicentre case-control study in the 28 French 'SIDS Centers'. Case selection was based on death labelled sudden infant death syndrome (SIDS) of an infant aged between 30 and 90 days. Three living controls were selected, matched for sex, gestational age and born immediately after the victim in the same maternity unit. RESULTS: We identified 114 cases of SUD aged between 30 and 90 days and 341 live controls matched for age and sex and born in the same maternity unit as the case. DTPP+/-Hib immunization did not increase the risk of SUD (OR 1.08) (95% CI 0.49, 2.36) in children under 3 months of age when adjusted for sleeping position, illness in the week before death, maternal tobacco consumption, birth weight, type of mattress, breastfeeding and sex. However, low birth-weight (6.53 [2.29, 18.9]), multiple birth (5.1 [1.76, 15.13]), no breastfeeding (1.77 [1.1, 2.85]), prone sleeping position (9.8 [5, 8, 18, 9]), soft mattress (3.26 [1.69, 6.29]), recent illness (3.44 [1.84, 6.41]) and parental smoking (1.74 [1.2, 2.96]) were confirmed as risk factors in early SIDS. CONCLUSIONS: DTPP+/-Hib immunization is not a risk factor for early SUD. In this population, we found the same risk factors as described for SIDS.

Prospective randomized multicenter comparison of high-frequency oscillatory ventilation and conventional ventilation in preterm infants of less than 30 weeks with respiratory distress syndrome.

BACKGROUND: Early use of high-frequency ventilation and exogenous surfactant is proposed as the optimal mode of ventilatory support in infants with respiratory distress syndrome. In very premature infants, we tested the hypothesis that high-frequency versus conventional ventilation could decrease exogenous surfactant requirements and improve pulmonary outcome, without altering the complication rate, including that of severe intraventricular hemorrhage. METHODS: Preterm infants with a postmenstrual age of 24 to 29 weeks, presenting with respiratory distress syndrome were randomly assigned to high-frequency oscillatory ventilation (lung volume recruitment strategy) or conventional ventilation. RESULTS: Two hundred seventy-three infants were enrolled. One hundred fifty-three had a postmenstrual age of 24 to 27 weeks, and 143 had a birth weight /=2 instillations of exogenous surfactant (30% vs 62%; odds ratio:.27; 95% confidence interval:.16-.44) and no difference in pulmonary outcome. The incidence of severe intraventricular hemorrhage was 24% in the high-frequency group and 14% in the conventional ventilation group (adjusted odds ratio: 1.50; 95% confidence interval:.68-3.30). CONCLUSION: Early use of high-frequency oscillatory ventilation in very premature infants decreases exogenous surfactant requirements, does not improve the pulmonary outcome, and may be associated with an increased incidence of severe intraventricular hemorrhage.

Opposing effects of excitatory amino acids on chick embryo spinal cord motoneurons: excitotoxic degeneration or prevention of programmed cell death.

Acute administration of a single dose of NMDA on embryonic day (E) 7 or later induces a marked excitotoxic injury in the chick spinal cord, including massive necrotic motoneuron (MN) death. When the same treatment was performed before E7, little, if any, excitotoxic response was observed. Chronic treatment with NMDA starting on E5 prevents the excitotoxic response produced by a later "acute" administration of NMDA. Additionally, chronic NMDA treatment also prevents the later excitotoxic injury induced by non-NMDA glutamate receptor agonists, such as kainate or AMPA. Chronic NMDA treatment also reduces normal MN death when treatment is maintained during the period of naturally occurring programmed cell death (PCD) of MNs and rescues MNs from PCD induced by early peripheral target deprivation. The trophic action of chronic NMDA treatment appears to involve a downregulation of glutamate receptors as shown by both a reduction in the obligatory NR1 subunit protein of the NMDA receptor and a decrease in the kainate-induced Co(2+) uptake in MNs. Both tolerance to excitotoxicity and trophic effects of chronic NMDA treatment are prevented by the NMDA receptor antagonist MK-801. Additionally, administration of MK-801 alone results in an increase in MN PCD. These data indicate for the first time that early activation of NMDA receptors in developing avian MNs in vivo has a trophic, survival-promoting effect, inhibiting PCD by a target-independent mechanism that involves NMDA receptor downregulation.

Activation and desensitization by cyclic antidepressant drugs of alpha2-autoreceptors, alpha2-heteroreceptors and 5-HT1A-autoreceptors regulating monamine synthesis in the rat brain in vivo.

The effects of antidepressant drugs on the synthesis of noradrenaline and serotonin (5-HT) were assessed using the accumulation of 3,4-dihydroxyphenylalanine (dopa) and 5-hydroxytryptophan (5-HTP) after decarboxylase inhibition as a measure of the rate of tyrosine and tryptophan hydroxylation in the rat brain in vivo. Three inhibitory synthesis-modulating receptors were investigated simultaneously: the alpha2C-autoreceptor modulating dopa/noradrenaline synthesis, and the alpha2A-heteroreceptor and 5-HT1A-autoreceptor modulating 5-HTP/5-HT synthesis. Acute treatment (2 h, i.p.) with desipramine (1-10 mg/kg), protriptyline (0.3-10 mg/kg) and nisoxetine (3-10 mg/kg), selective NA reuptake blockers, dose-dependently decreased dopa synthesis in cortex (15%-40%) and hippocampus (20%-53%). Fluoxetine (1-10 mg/kg) and zimelidine (1-10 mg/kg), selective 5-HT reuptake blockers, did not alter dopa synthesis. Fluoxetine and zimelidine dose-dependently decreased 5-HTP synthesis in cortex (14%-43%) and hippocampus (27%-54%). Desipramine and protryptyline did not alter 5-HTP synthesis in cortex but in hippocampus it was decreased (36%). Repeated desipramine (10 mg/kg for 1-21 days) or fluoxetine (3 mg/kg for 3-21 days) treatment resulted in a time-dependent loss in their ability to decrease dopa or 5-HTP synthesis. Desipramine (1-21 days) did not alter 5-HTP synthesis in cortex, but in hippocampus it was decreased (21%-37%, days 1-14) followed by recovery to control values (day 21). Fluoxetine (3-21 days) did not alter brain dopa synthesis. To further assess the desensitization of alpha2C-autoreceptors, alpha2A-heteroreceptors and 5-HT1A autoreceptors regulating the synthesis of dopa/NA or 5-HTP/5-HT after chronic desipramine and fluoxetine, the effects of clonidine (agonist at alpha2-auto/heteroreceptors) and 8-OH-DPAT (agonist at 5-HT1A-autoreceptors) were tested. In saline-treated rats, clonidine (1 mg/kg, 1 h) decreased dopa and 5-HTP synthesis in cortex (58% and 54%) and hippocampus (54% and 42%). In desipramine-treated rats (10 mg/kg, 21 days), but not in fluoxetine-treated ones (3 mg/kg, 14 days), the effect of clonidine was attenuated in cortex (12% and 18%) and only for dopa synthesis in hippocampus (31%). In saline-treated rats, 8-OH-DPAT (1 mg/kg, 1 h) decreased 5-HTP synthesis in cortex (63%) and hippocampus (75%). In fluoxetine-treated rats, but not in desipramine-treated ones, this inhibitory effect was markedly attenuated in cortex (26%) and hippocampus (9%). These findings indicate that acute treatment with cyclic antidepressant drugs results in activation of inhibitory alpha2C-autoreceptors, alpha2A-heteroreceptors and/or 5-HT1A-autoreceptors regulating the synthesis of dopa/NA and/or 5-HTP/5-HT in brain, whereas chronic treatment with these drugs is followed by desensitization of these presynaptic receptors.

Inhaled salbutamol and beclomethasone for preventing broncho-pulmonary dysplasia: a randomised double-blind study.

UNLABELLED: Early inflammatory lesions and bronchial hyperresponsiveness are characteristics of the respiratory distress in premature neonates and are susceptible to aggravation by assisted ventilation. We hypothesized that treatment with inhaled salbutamol and beclomethasone might be of clinical value in the prevention of bronchopulmonary dysplasia (BPD) in ventilator-dependent premature neonates. The study was double-blinded and placebo controlled. We studied 173 infants of less than 31 weeks of gestational age, who needed ventilatory support at the 10th postnatal day. They were randomised to four groups and received either placebo + placebo, placebo + salbutamol, placebo + beclomethasone or beclomethasone + salbutomol, respectively for 28 days. The major criteria for efficacy were: diagnosis of BPD (with score of severity), mortality, duration of ventilatory support and oxygen therapy. The trial groups were similar with respect to age at entry (9.8-10.1 days), gestational age (27.6-27.8 weeks), birth weight and oxygen dependence. We did not observe any significant effect of treatment on survival, diagnosis and severity of BPD, duration of ventilatory support or oxygen therapy. For instance, the odds-ratio (95% confidence interval) for severe or moderate BPD were 1.04 (0.52-2.06) for inhaled beclomethasone and 1.54 (0.78-3.05) for inhaled salbutamol. CONCLUSION: This randomised prospective trial does not support the use of treatment with inhaled beclomethasone, salbutamol or their combination in the prevention of BPD in premature ventilated neonates.

Effects of excitatory amino acids on neuromuscular development in the chick embryo.

To investigate the presumptive role of excitatory amino acids (EAAs) in the regulation of normally occurring motoneuron (MN) death, chick embryos were treated with the glutamate receptor antagonists dizocilpine maleate and 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]quinoxaline-7-sulfonamide disodium. Both failed to alter the number of surviving MNs at the end of the critical period of programmed cell death. However, treatment with 3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid, a competitive N-methyl-D-aspartic acid (NMDA) receptor antagonist, was able to rescue a significant number of MNs from death. Treatment with several EAA agonists induced extensive excitotoxic lesions in the spinal cord. MN degeneration induced by excitotoxins exhibited changes characteristic of necrosis rather than apoptosis. However, when either 0.5 or 1 mg of NMDA was applied acutely on embryonic day (E) 7, about 50% of treated embryos failed to exhibit NMDA-induced excitoxicity but rather showed a clear reduction in the number of pyknotic MNs. This apparent neuroprotective effect of NMDA was also observed in a subset of embryos chronically treated with NMDA, in which an excessive number of MNs was detected when examined on E9. Surprisingly, in the same experiment other embryos showed either normal or highly reduced MN numbers. Embryos with motoneuronal depletion induced by NMDA also showed a delayed impairment of later neuromuscular development with the appearance of degenerative changes in surviving MNs and apoptosis of skeletal muscle cells. Because some of the alterations reported here are similar to those described in MN diseases, our experimental model may be useful for gaining insights into the mechanisms that control both developmentally regulated and pathological MN death.

Effect of uncomplicated chronic hypertension on the risk of small-for-gestational age birth.

This study assesses the effect of chronic hypertension on fetal growth. A cohort of 1,938 pregnant women attending five prenatal clinics in France between August 1991 and May 1993 were enrolled in a prospective study. Chronic hypertension was defined according to blood pressure at enrollment and past history, and cases complicated by preeclampsia were excluded. Adjusted odds ratios of small-for-gestational age birth were estimated by logistic regression. The independent effect of chronic hypertension on mean birth weight was examined through multiple linear regression analysis adjusting for gestational age at delivery and potential confounders. Uncomplicated chronic hypertension was associated with an increased risk of small-for-gestational age birth. Odds ratios increased with age. In women over age 30 years, the association was strong (adjusted odds ratio = 8.5, 95% confidence interval 2.9-24.5). Multiple linear regression showed that mean birth weight was 161 g (95% confidence interval 66-256 g) less in women with chronic hypertension compared with normotensive women. The authors conclude that mean birth weight is reduced and the risk of small-for-gestational age birth is increased in uncomplicated chronic hypertension compared with normotensive pregnancies. Results further suggest that the magnitude of this association increases with age.

Umbilical artery Doppler velocimetry in unselected and low risk pregnancies: a review of randomised controlled trials.

OBJECTIVE: Meta-analysis of data from controlled trials performed in populations at high risk have shown that umbilical artery Doppler velocimetry (umbilical Doppler) can reduce perinatal mortality. The individual published trials among unselected or low risk populations have found no beneficial effect. Our objective was to evaluate the effect of routine use of the umbilical Doppler in unselected or low risk pregnancies by reviewing all published and unpublished randomised controlled trials. STUDY DESIGN: Systematically reviewing published and unpublished trials, we selected trials for the overview only if they were completed randomised trials of umbilical Doppler in unselected or low risk pregnancies. Of the seven trials examined, three were not included in the meta-analysis because of methodological problems or because they did not meet the inclusion criteria. We therefore included four trials: two in unselected and two in low risk populations. These four groups were comparable in their degree of maternal and perinatal risk. Using the Mantel-Haenzel statistical method and pooling the data, we considered 11,375 women in the meta-analysis. RESULTS: Systematic use of the Doppler umbilical artery velocimetry had no statistically significant effect on perinatal deaths in unselected populations (odds ratio [OR] 1.28; 95% confidence interval [CI] 0.61-2.67), low risk populations (OR 0.51; 95% CI 0.20-1.29) or overall for the four trials (OR 0.90; 95% CI 0.50-1.60); nor was there any significant effect on stillbirths (global OR 0.94; 95% CI 0.42-1.98). However, the number of participants remain insufficient and further information is required to arrive at a definite conclusion on the absence of effect. The meta-analysis showed no significant difference between the Doppler groups and the control groups for antenatal hospitalisation, obstetric outcome or perinatal morbidity. CONCLUSION: Based on the results of the published trials, routine use of the umbilical Doppler cannot be recommended.

The alpha 2-adrenoceptor antagonist idazoxan is an agonist at 5-HT1A autoreceptors modulating serotonin synthesis in the rat brain in vivo.

The in vivo effects of the alpha 2-adrenoceptor idazoxan, rauwolscine and phentolamine on alpha 2-auto/heteroreceptors and 5-HT1A autoreceptors modulating the synthesis of dopa/noradrenaline and 5-HTP/serotonin were assessed in rats, using the accumulation of dopa and 5-HTP after decarboxylase inhibition as a measure of the rate of tyrosine and tryptophan hydroxylation. The acute administration of idazoxan (0.1-40 mg/kg) induced a pronounced dose-dependent increase in the synthesis of dopa in the cerebral cortex (22-86%) and hippocampus (8-80%), as a consequence of the powerful blockade of alpha 2-autoreceptors. However, idazoxan did not increase the synthesis of 5-HTP in these brain regions, as it would have been expected by the concurrent blockade of alpha 2-heteroreceptors on serotonergic terminals. Instead, idazoxan decreased the synthesis of 5-HTP in the cerebral cortex (13-33%) and hippocampus (25-48%), suggesting that these inhibitory effects were mediated through activation of 5-HT1A autoreceptors. Similar results were obtained for rauwolscine. Pre-treatment of rats with the selective 5-HT1A receptor antagonist WAY100135 (10 mg/kg) fully antagonized the inhibitory effects of idazoxan (10 mg/kg) on 5-HTP synthesis, but it did not prevent the stimulatory effects of idazoxan on dopa synthesis. The results indicate that idazoxan is a potent and specific agonist at 5-HT1A autoreceptors modulating brain serotonin synthesis in vivo.

Alpha 2-autoreceptors and alpha 2-heteroreceptors modulating tyrosine and tryptophan hydroxylase activity in the rat brain in vivo: an investigation into the alpha 2-adrenoceptor subtypes.

The subtype determination of auto- and hetero-alpha 2-adrenoceptors modulating the synthesis of noradrenaline (NA) and serotonin (5-HT), respectively, was assessed using the accumulation of 3,4-dihydroxyphenylalanine (dopa) and 5-hydroxytryptophan (5-HTP) after decarboxylase inhibition as a measure of the rate of tyrosine and tryptophan hydroxylation in the rat brain in vivo. In the cerebral cortex and hippocampus, Org 3770 (non-selective alpha 2-adrenoceptor antagonist, 0.5-10 mg/kg, i.p.) increased (43%-58%) and clonidine (non-selective alpha 2-adrenoceptor agonist, 1 mg/kg) decreased (37%-49%) the synthesis of dopa. Also the antagonist ARC 239 (alpha 2B/C selective, 5-40 mg/kg) increased the synthesis of dopa in cortex (39%-46%) and hippocampus (17%-85%). In contrast, the antagonist BRL 44408 (alpha 2D selective, 1-10 mg/kg) did not increase the synthesis of dopa in cortex, and increased it modestly in hippocampus only. The agonist guanoxabenz (alpha 2B/C selective, 0.03-3 mg/kg) decreased the synthesis of dopa in both brain regions (20%-65%), whereas the agonist oxymetazoline (alpha 2D selective, 0.1-3 mg/kg) failed to do so. These results indicated that the alpha 2-autoreceptors that modulate the synthesis of dopa/NA are probably associated with the alpha 2B/C-subtypes. In cortex and hippocampus, clonidine decreased (35%-53%) the synthesis of 5-HTP but Org 3770 failed to induce the opposite effect (except the 2 mg/kg dose in cortex). BRL 44408 markedly increased the synthesis of 5-HTP in cortex (113%-148%) but not in hippocampus. Similarly, also ARC239 increased the formation of 5-HTP in cortex (36%-48%) but not in hippocampus, where it was decreased (30%-55%). Oxymetazoline decreased the synthesis of 5-HTP in hippocampus (28%-30%) but failed to do so in cortex. Guanoxabenz in the low dose range (0.03-0.3 mg/kg) did not decrease the synthesis of 5-HTP in any brain region. These results indicated that the alpha 2-heteroreceptors that modulate the synthesis of 5-HTP/5-HT may well be different from the proposed alpha 2B/C-autoreceptors modulating the synthesis of dopa/NA. These alpha 2-heteroreceptors appear to be associated with the alpha 2D-subtype.

Porcine surfactant replacement therapy in newborns of 25-31 weeks' gestation: a randomized, multicentre trial of prophylaxis versus rescue with multiple low doses. The French Collaborative Multicentre Study Group.

The aim of the study was to determine if prophylaxis with multiple low doses of porcine surfactant would increase survival, without bronchopulmonary dysplasia, compared with rescue therapy, for respiratory distress syndrome in newborns of 25-31 weeks' gestation. Compared with rescue therapy (n = 122), prophylaxis (n = 134) decreased the need for oxygenation and ventilatory support within 3-72 h. It did not, however, increase survival without bronchopulmonary dysplasia (60% versus 46%) (odds ratio (OR) = 1.53, 95% confidence interval (CI) = 0.90-2.61). Furthermore, prophylaxis decreased the incidence of severe peri-intraventricular haemorrhage (3% versus 16%) (OR = 0.28, 95% CI = 0.09-0.84) and retinopathy of prematurity (2% versus 11%) (OR = 0.18, CI = 0.04-0.78). We conclude that prophylaxis did not increase survival without bronchopulmonary dysplasia. The decreased incidence of severe peri-intraventricular haemorrhage and retinopathy of prematurity after prophylaxis requires further study.