Supersensitivity of human metabotropic glutamate 1a receptor signaling in L929sA cells

The effect of antagonist pretreatment on the signaling properties of the human metabotropic glutamate 1a (hmGlu1a) receptor was examined in stably transfected L929sA cells. Pre-exposure of hmGlu1a receptor-expressing cells to the mGlu1 receptor antagonists (S)-4-carboxy-3-hydroxyphenylglycine and 7-(hydroxyimino)cyclo-propa[b]chromen-1a-carboxylate ethyl ester dramatically enhanced subsequent glutamate-induced phosphoinositide hydrolysis and intracellular [Ca(2+)] rise. We found clear indications that the antagonist-mediated enhancement of glutamate-evoked mGlu1a receptor signaling is caused by the development of mGlu1a receptor supersensitivity: the potency of glutamate was increased by 3-fold after 24 h antagonist pretreatment and the potency of the antagonists was significantly decreased in antagonist-pretreated cells. The kinetic profile of the antagonist-mediated enhancement showed that the maximal increase in intracellular [Ca(2+)] was already reached after 30-min pretreatment, suggesting that de novo receptor synthesis is not involved in the process of mGlu1a receptor supersensitization. Glutamate-mediated phosphoinositide hydrolysis increased up to 24 h after antagonist treatment. Although it seemed likely that the hmGlu1a receptor could desensitize after activation by endogenously present glutamate, removal of glutamate from the extracellular medium with GPT resulted in a much smaller enhancement of glutamate responsiveness. Moreover, the magnitude of antagonist-mediated receptor supersensitivity was much larger than the magnitude of agonist-induced receptor desensitization. These results suggest that antagonist-evoked mGlu1 receptor supersensitivity is not merely the result of a blockade of agonist-induced desensitization. Finally, we found that antagonist pretreatment doubled the amount of receptors at the cell surface. Our findings are the first lines of evidence that prolonged antagonist treatment can supersensitize the hmGlu1a receptor. In view of the potential therapeutic application of mGlu1 receptor antagonists, it will be important to know whether these phenomena occur in vivo.     

The chemical UV-filter 3-benzylidene camphor causes an oestrogenic effect in an in vivo fish assay

Chemical UV-filters are used in sun protection products and various kinds of cosmetics. The lipophilic chemical UV-filter 3-benzylidene camphor was investigated for its capability to cause vitellogenin induction, possibly via oestrogen receptor binding, in a well-established in vivo fish assay (juvenile rainbow trout, Oncorhynchus mykiss, vitellogenin ELISA). A clear relationship was demonstrated between the dose of injected 3-benzylidene camphor and the concentration of plasma vitellogenin with a 105-times induction from 68 mg 3-benzylidene camphor /kg/injection and above compared to the control vitellogenin level. The relationship between the injected dose of 3-benzylidene camphor and the percent of responding fish (vitellogenin) was evaluated by logistic regression analysis and effective dose-values (ED-values) were determined. ED10, ED50 and ED90 of 3-benzylidene camphor after 6 days (2 injections) were 6.4, 16 and 26 mg/kg/ injection, respectively. These ED-values place 3-benzylidene camphor among the more potent xenooestrogens discovered to date and necessitates investigations on the distribution, concentration, persistence and bioaccumulation of 3-benzylidene camphor and other UV-filters in nature.     

Alpha-synuclein and presynaptic function: implications for Parkinson's disease

This article focuses on alpha-synuclein's role in normal and pathological axonal and presynaptic functions and its relationship to Parkinson's disease. It is not possible to mention all the contributions to aspects of this area. Readers interested in alpha-synuclein's relation to aggregation, Lewy lesions, and pathological modifications are referred to the many reviews (see Goldberg and Lansbury 2000; Galvin 2001a; Goedert 2001).     

EEG measurements by means of radiotelemetry after intracerebroventricular (ICV) cannulation in rodents

The utility of implanted radiotelemetry transmitters for the measurement of electroencephalogram (EEG), locomotor activity, body temperature and cardiovascular parameters has been well documented. This paper focuses on the methodology, of combining radiotelemetry with intracerebroventricular (ICV) cannulation. The two excitatory neuropeptides, orexin-A and orexin-B, can only be given by ICV injection, therefore we examined their effects on the normal sleep-wake cycle of rodents in the present study. The effects of orexins on sleep architecture have been extensively studied in tethered models demonstrating increased levels of wakefulness. In this study, both orexin neuropeptides, especially orexin-A, increase wakefulness within the first sleep period followed by an increase in slow wave (SW) sleep and paradoxical sleep (PS), towards the end of a 5-h recording period which may be a rebound phenomena. The present study has demonstrated that ICV cannulation can be used effectively in studying the effects of pharmacological agents on the sleep-wake cycle of rodents by measuring EEG and EMG by radiotelemetry.     

Illness-related behaviour and utilization of oral health services among adult city-dwellers in Burkina Faso: evidence from a household survey

Background  

In sub-Saharan Africa, the availability and accessibility of oral health services are seriously constrained and the provision of essential oral care is limited. Reports from the region show a very low utilization of oral health care services, and visits to dental-care facilities are mostly undertaken for symptomatic reasons. The objectives of the present study were to describe the prevalence of oral symptoms among adults in Ouagadougou, capital city of Burkina Faso and the use of oral health services and self-medication in response to these symptoms and to measure the associations between predisposing, enabling and needs factors and decisions to seek oral health care.     

EFFECT OF MODERATE CARBON-MONOXIDE EXPOSURE ON FETAL DEVELOPMENT

The effect of moderate carbon-monoxide exposure (180 p.p.m. and 90 p.p.m. carbon monoxide in atmospheric air) on fetal development was studied in rabbits. Exposure to 180 p.p.m. carbon monoxide (16-18% carboxyhæmoglobin) during pregnancy resulted in a 20% decrease of birth-weight, and a neonatal mortality of 35% as against 1% in the control group. Exposure to 90 p.p.m. carbon monoxide (8-9% carboxyhæmoglobin) had a less pronounced effect. In women there was a negative correlation between birth-weight and carboxyhæmoglobin concentration (p<0·05). The results indicate that carbon monoxide in tobacco smoke might be responsible for the reduced birth-weight of babies whose mothers smoked during pregnancy.     

Adrenal cortex and avitaminosis A: II. Complementary in vivo and in vitro studies on inhibition of corticoidogenesis in moderate deficiency in the rat. III. In vivo criterion for activity: glycemic response to ACTH]

II. We set up new experiments, both in vivo and in vitro, in order to explain the impaired adrenal response to stress, of vitamin A middly deficient, male Wistar rats. Injections of progesterone, 6 mg s.c. or i.p. every other day, did not improve the deficiency curse in our rats. In agreement with these results, we have found that, in vitro, the delta5-3beta hydroxysteroide dehydrogenase is not altered in deficient glands; we confirmed it histochemically. On the other hand, the 11beta and 18 hydroxylase activities are lowered in vitro (and presumably in vivo). When conditions of hyperactivity are established, the conversion of deoxycorticosterone (DOC) to 18 OH-DOC or to corticosterone is reduced and DOC accumulates in the incubation medium. III. When middly deficient intact rats are injected with ACTH i.p. (2UI p. 100 g of body weight), only some of them are found to have normoglycemia or hyperglycemia, 45 or 60 minutes after injection, as the controls do. Many deficient rats become hypoglycemic as the adrenalectomised animals do. We think that the changes in the glycemia after ACTH injection could be used to discriminate between the more or less deficient rats.     

Pharmacokinetics and D2 receptor occupancy of long-acting injectable risperidone (Risperdal Consta) in patients with schizophrenia

Thirteen patients with schizophrenia received injections of 25, 50, or 75 mg of long-acting risperidone every 2 wk. Brain D2 receptor occupancy was assessed with [11C]raclopride 2 wk after the last (fifth) injection (day 71) in seven subjects and 2 wk after the third injection (day 44) in one subject. Stable plasma concentrations were reached after the third injection and steady-state concentrations of the active moiety (risperidone + 9-hydroxyrisperidone) after the fourth injection. Steady-state plasma concentrations were maintained for 4-5 wk after the last injection and then declined rapidly. After injections of 25, 50 and 75 mg on day 44 or day 71, D2 receptor occupancy ranged from 25-48%, 59-83% and 62-72% respectively, while plasma active-moiety levels ranged from 4.4-8.8, 15.0-31.1 and 22.5-26.3 ng/ml respectively. The results indicate that brain D2 receptor occupancy at steady state after injections of long-acting risperidone was in the range found in patients effectively treated with 2-6 mg of oral risperidone.     

Highly efficient and reliable chemically assisted enucleation method for handmade cloning in cattle

The purpose of the present study was to find an efficient and reliable chemically assisted procedure for enucleation related to the handmade cloning (HMC) technique. After in vitro maturation oocytes were incubated in 0.5 microg mL(-1) demecolcine for 2 h. Subsequently, zonae pellucidae were digested with pronase, and one-third of the cytoplasm connected to an extrusion cone was removed by hand using a microblade. The remaining two-thirds were used as recipients for HMC, and reconstructed and activated embryos were cultured for 7 days. The time-dependent manner of the development of extrusion cones, the efficiency (oriented bisection per oocyte; 94%), reliability (success per attempted enucleation; 98%), and the blastocyst per reconstructed embryo rates (48%) were measured. Ultrastructural analyses demonstrated that demecolcine treatment resulted in disoriented and haphazardly orientated microtubules. The general ultrastructure of the oocyte organelles, however, appeared to be unaltered by the treatments. Considering that no oocyte selection based on polar body presence was performed, this system seems to be more efficient and reliable than any other enucleation method. Moreover, expensive equipment (inverted fluorescence microscope) and a potentially harmful step (staining and ultraviolet illumination) can be eliminated from the HMC procedure without compromising the high in vitro efficiency.