[Mycetoma in Mauritania: species found, epidemiologic characteristics and country distribution. Report of 122 cases]. A two year investigation conducted at Nouakchott's Centre Hospitalier National supplied 122 histologically confirmed cases of mycetoma. 75% of those were of mycotic origin, of which the most common species was Madurella mycetomatis. The mean ages were 28 to 29 years at the beginning of the disease and 38 years at the time of examination. The duration of the evolution was between 6.3 and 8.4 years. Madurella mycetomatis seem to appear at a lower age (mean age beginning at 21.2 years, consultation at 27). Patients with bone involvement are older than others (38 versus 28 years). There is a statistically significant relation between spontaneous pain and bone involvement.


[Paracoccidioidomycosis assessed with 4 isotopic studies]. Paracoccidioidomycosis (south-american blastomycosis) is a consumptive granulomatous disease that is found in all Latin-America. The ethiological agent is a fungus (Paracoccidiosis brasiliensis) and mechanisms of disease transmission are not clear. Affection is frequent in the lungs, liver, spleen, ganglia, oropharynx and brain. Disease's response to therapy (Amphotericin, Ketoconazole) is good but recurrence is frequent. We have studied 12 patients with gallium-67, bone scintigraphy, bone marrow scintigraphy and lymph-scintigraphy. Six of these patients were re-studied after variable time of therapy. Gallium-67 accumulated in all patients's lungs. Other abnormal areas included ganglia, liver, spleen and bone. Bone scintigraphy was abnormal in all bone sites that were abnormal at the gallium study. Bone marrow scintigraphy showed expansion to the periphery in 6/12 cases and lymph nodes accumulated the radiopharmaceutical in 5 patients. Patients re-studied after the beginning of therapy presented decreased degrees or normalization of the radio-pharmaceutical's uptake (gallium and MDP), a regression of the bone marrow periphery expansion (bone marrow scintigraphy) and normalization of the lymph node aspects. These four radioisotopical studies may be useful for staging and for therapy follow-up. The sensitivity of the method is greater than the one noted for the conventional radiological studies.


[Keloid paracoccidioidomycosis assessed with gallium]. This rare mycosis is caused by Paracoccidiodis loboii and is found mainly among indians of the Amazon forest. It is a cutaneous disease. No visceral affection was ever described, though generalized skin disease has been reported. One case of cheloid paracoccidioidomycosis was studied with gallium-67. The radiopharmaceutical has accumulated in the affected area and in a lymph node, although adenomegaly be rare. Degree of accumulation was discrete/moderate but the patient had been under irregular treatment for 10 years. As no laboratorial data are evaluable for this affection, gallium-67 may represent a means of disease extent and therapy efficacy evaluation.


[Genital infections with Chlamydia trachomatis in pregnant women in New Caledonia]. Chlamydia trachomatis was researched from 400 pregnant women. 76 of them were found infected and 129 carried serological signs of infection. We purpose a detection at the end of pregnancy interesting women with sexually transmitted diseases background and those with serological positive results (greater than 1/64). Then, treatment of the mother will be an useful method of prevention against neonatal infection.


[Seroepidemiologic study of sexually transmitted diseases caused by Chlamydia trachomatis in Casablanca (Morocco)]. Screening 579 sera specimens obtained in Casablanca (Morocco) we have performed a retrospective seroepidemiological survey with regard to the prevalence of Chlamydia trachomatis involved in genital tract infections. The surveyed populations are divided into 3 groups: 177 patients affected with sexually transmitted diseases (STD), 319 reference patients, 83 maghrebian patients affected with STD but living in France. The study was performed by the indirect microimmunofluorescence technique. The prevalence of Chlamydia trachomatis ranging from 50% in the moroccan reference groupe to about 70% among patients with STD both in Morocco and Bordeaux indicates that Chlamydia trachomatis takes a significant part in STD observed in moroccan populations.


[Epidemiologic surveillance of tuberculosis in French Polynesia between 1960 and 1990]. A monitoring and prevention policy for tuberculosis has been implemented in French Polynesia in 1960. Between 1960 and 1990, case-notification rate decreased dramatically from 568 to 30 per 100,000 with an average annual decrease rate of 9.1%. Cumulative case-notification rates calculated over the 30-year period for the 5 archipelagoes constituting the territory vary significantly: the highest rate has been observed in Gambier Islands. Sex ratio for the whole period was 1.1. The annual incidence rate, according to age-groups has been computerized using actuarial method. A cohort-effect is observed: given an age of individuals, the incidence rate becomes lower when shifting from an older cohort to a younger one. Also, an age-effect is to be emphasized: there is a decreasing trend of the rate in each cohort when the age of the individuals amid the cohort increases. The proportion of smear-positive tuberculosis cases increased from 38% in 1976 to 54% in 1990. A 0.57% annual risk of infection (ARI) has been estimated for 1977, and a 0.41% ARI for 1989. Tuberculosis mortally rate is documented since 1985, and varies between 10.8 and 2.1 per 100,000 since that year.


[Resistance of Vibrio cholerae 0:1 to the 0/129 compound and multiresistance to antibiotics]. The authors present a study of the in vitro susceptibility to O/129 compound and usual antibiotics of 29 strains of V. cholerae O:1 biotype El Tor isolated during epidemics in miscellaneous countries over the world from 1982 to 1991. Several identical isolates from the same epidemic are represented by one strain. Susceptibility testing by diffusion method and MICs by agar dilution method are used. The data show that the resistance to O/129 compound is often associated with the resistance to usual antibiotics such as trimethoprim, sulphonamides, chloramphenicol, ampicillin and tetracycline. This resistance to the vibrostatic compound leads to a double problem of diagnosis and therapy. The nitrofuranes derivatives and tiliquinol-tilbroquinol association, an intestinal antiseptic, are the most active antimicrobial agents as well on the strains O/129 sensitive as on the strains O/129 resistant.


[Involvement of cellular immunity in pathology. Neuromalaria]. Murin Cerebral Malaria (MCM) with Plasmodium berghei ANKA and the CBA/Ca mice is the result of an immunopathological process. An overproduction of TNF is implicated in its pathogenesis. Recent datas concerning TNF production during the course of Plasmodium vinckei vinckei infection, and analysis of relationships between MCM and Experimental Allergic Encephalomyelitis (EAE) raise the hypothesis of the involvement of an auto-immune process in the murin disease. The role of cellular immunity in human cerebral malaria remains obscure. Cytokines could majore adherence of parasitized red blood cells to cerebral endothelial cells.


[Pathophysiology of malaria. The current issue]. Patho-physiology of malaria is often presented with very old, rudimentary and not scientifically proved explanations. In fact, the mechanisms involved in malaria and mainly in cerebral malaria are very complex and, until now, partly unknown. Four hypotheses (sludging, modifications or blood-brain barrier permeability, mechanical or immunological phenomena) have to be considered. Several cytokines and mainly TNF seem to play the major role but other parameters are also essential in relation with Plasmodium (strain characteristics) or with the patient (HLA groups, immunological response). All technologies (immunology, iso-enzymology, molecular biology) bring new and important informations but, at the same time, make the issue ever more complex. Nevertheless, patho-physiology remains one of the most interesting aspects of malaria, both from a fundamental and a practical point of view, as fighting against this disease will probably become easier when we understand its mechanisms better.


[Visceral leishmaniasis in Cameroon. Report of various cases and clinical study in the region of Kousseri, far-north of Cameroon]. The far-North province of Cameroon is highly infested with cutaneous leishmaniasis. From October 1987 to January 1988, a prospective study was done in this region (specifically in Kousseri) to diagnose cases of visceral leishmaniasis (VL). Out of 120 patients examined, 46 presenting symptoms related to VL were retained. Diagnosis of VL was confirmed serologically and/or through the presence of leishmanias on bone marrow slides from 9 patients. A retrospective study was also carried out in the pediatric and internal medicine units of the university teaching hospital and the Central Hospital in Yaoundé; 3 pediatric cases of VL were found. No significant difference as regards age and sex of these patients was recorded.


[Canine leishmaniasis in the region of Enfidha (Central Tunisia). Assessment of seroprevalence with direct agglutination (DAT) and indirect immunofluorescence (IFAT)]. In order to assess canine leishmaniasis prevalence rate in Enfidha area, considered to be the most important kala-azar focus in Sousse Governorate, a serological survey was carried out in 6 localities. 265 sera were examined by DAT and IFAT. 16 (6.03%) showed positive results for anti-leishmania antibodies with significant variations according to the locality. A fairly DAT-IFAT good correlation was observed.


[Characterization and evolution of blood cryoglobulin in visceral leishmaniasis]. The evolution of immunological parameters in mediterranean visceral leishmaniasis reveal a type III mixed cryoglobulin with rhumatoid factor activity and antileishmania antibodies. This cryoglobulin follows the clinical signs of the disease and disappear under treatment. The authors insist on the transitory character of this cryoglobulin which is in relation with polyclonal stimulation of the immune system by parasitical infection.


[Cryptosporidiosis in Ivorian children form Yopougon]. Coprological survey involving 250 ivorian children with diarrhoea was done to evaluate the incidence of cryptosporidiosis. Crypstosporidium sp. was found in 10.4% of subjects. 76.9, 57.7 and 19.2% Cryptosporidium positive children had profuse diarrhoea, fever and pulmonary symptoms respectively. In Ivory Coast, children diarrhoeas due to cryptosporidiosis are quantitatively important.


[Peritoneal dirofilariasis fortuitously disclosed]. Peritoneal dirofilariasis probably due to Dirofilaria repens is reported from a 52 years old woman. The parasite discovery is fortuitous during a surgical cure of intestinal adenocarcinoma. The patient lives in Provence and Corsica (Southern France), an endemic region. The authors present the histological aspects of the parasite.


[First observations on the main plasma parameters of oxidative stress in homozygous sickle cell disease]. The present study report 7 cases of sickle homozygous disease which have been analysed using markers of the oxidative-stress, 26 african male subjects were studied: 7 Hb SS subjects (age: m = 20) and 19 control subjects (Hb AA, age: m = 40). Plasma concentrations of F-MDA, T-MDA, TBARS, alpha tocopherol, retinol and beta carotene were measured. Plasma MDA and TBARS mean levels increased in sickle homozygous patients more than in controls. However, only TBARS mean concentrations were significantly increased between patients and controls: TBARS: 4.14 +/- 1.49 nMol/ml for Hb SS versus 2.10 +/- 1.21 nMol/ml for Hb AA (P less than 0.005). Vitamin A and vitamin E concentrations were significantly lower in Hb SS than in Hb AA. Beta carotene was significantly increased in patients vs controls. The significant increase of TBARS explains the great importance of the oxidative damage, whereas the significant decrease of vitamins A and E, may contribute, at least for a part, to maintain the autoxidation process or reveals its intensity in these patients.


[Overall analysis of structural factors responsible for the activity and toxicity of antiparasitic agents used in therapy]. A mini-data bank was constituted from the 73 french antiparasitic patent medicines and was implanted in microcomputer. A maximum of chemical, pharmaceutical, therapeutical and toxicological data were collected and treated by multivariate analysis. Exploitation of these informations processing allows to take out correlations between chemical structure toxicity, and specificity.


[Comparative efficacy of the use of CDC light traps and humans to sampling anopheles populations. Results obtained in the area of Bignona (Senegal)]. The simultaneous use for the purpose of comparison of human bait catch and light trap collection for sampling anopheles populations specially the Anopheles gambiae s.l. in the area of Bignona, Senegal shows that: light traps provide more information on the specific composition of anopheles fauna; the performance of light traps and that of human baits are comparable indoor during the period of high density of anopheles. Outdoor, light traps catch fourth less females A. gambiae. But the two methods always vary in the same pattern and identically reflect the density variations of the populations; there is no significant difference between the repletion status, the physiological age and the sporozoite index of females A. gambiae s.l. collected with each of the 2 methods. Light trap collection may therefore be considered a reliable sampling method for A. gambiae s.l. specially inside houses in some areas where this method can efficiently permit to complete the human bait catch for a malaria transmission study.


Predictive diagnosis of familial adenomatous polyposis with linked DNA markers: population based study. To evaluate the use of polymorphic DNA probes linked to the APC gene in the presymptomatic diagnosis of familial adenomatous polyposis. Four DNA probes were tested on an unselected population of patients at risk of familial adenomatous polyposis. The first 47 families notified to the West Midlands familial adenomatous polyposis register. Plus five families sent to our hospital as part of the West of Britain DNA consortium. The proportion of families and family members in whom DNA testing could be used to adjust the estimate of risk. Only 17 families on the register (containing 46% (74/162) of the population at risk) had a suitable pedigree structure for DNA analysis. DNA was analysed in 12 of these families plus the five families from the West of Britain consortium. At least one probe was informative in 27 of the 33 subjects born with 50% risk, but the most informative probe (pi 227) was the one with the highest recombination rate (10%). Flanking markers were informative in only four of the 33 subjects. These findings confirm the potential for accurate predictive diagnosis of familial adenomatous polyposis with polymorphic DNA probes, but such an approach is currently limited to about one third of affected families. A combined approach to presymptomatic diagnosis, which includes DNA testing and indirect ophthalmoscopy, is advocated.


Problem drug use in a central London general practice. To describe the profile of problem drug users presenting in general practice and to determine whether they can be cared for in general practice. Study of consultations by problem drug users. Central London general practice. 150 problem drug users presenting over two years. Stopping drug use, alterations in lifestyle, obtaining paid work, and stopping drug related crime. Of the 150 patients, 111 were men and 39 women, and 106 were unemployed. 121 patients used heroin, 112 of whom injected. 145 patients accepted a methadone reduction programme and 81 completed it. A further 25 were stabilised on reducing doses of methadone, until places became available for them as inpatients at drug dependency units or rehabilitation projects. Withdrawal programmes for patients who misuse drugs can be successfully managed in general practice.


Risk of HIV infection from transfusion with blood negative for HIV antibody in a west African city. To estimate the risk of infection with HIV (HIV 1 or HIV 2, or both) from transfusion of a screened unit of blood in a high prevalence area in west Africa. Retrospective cohort study for January-July 1991. National Blood Transfusion Centre, Abidjan, Côte d'Ivoire. Repeat donors (5831 units of blood) and first time donors (5076 units) in the first five months of 1991. Prevalence and estimated incidence of HIV infection in repeat and first time donors; estimated rate of potentially infected, HIV antibody negative units; and rate of (false negative) potentially infected units assuming a laboratory test sensitivity of 99%. Overall HIV prevalence was 11.0% in first time donors and 2.1% in repeat donors. In the first seven months of 1991, 29 HIV antibody positive (27 HIV 1, 1 HIV 2, 1 dually reactive) donors with a seronegative unit of blood earlier in the year were identified; 26 had donated blood eight weeks or less before their estimated dates of seroconversion and may have been infectious (minimum rate 26/5831 (4.5/1000 potentially infected units)). Estimated incidence of infection in repeat donors was 1.2-2.5%. Laboratory test insensitivity would result in an estimated 1.1/1000 false negative units from first time donors and 0.2/1000 units from regular donors. The overall rate of potentially infected units (all donors, seroconversions, and errors) was estimated at 5.4-10.6/1000. The risk of HIV infection from a single unit of blood remains substantial (5.4-10.6/1000 units). To prevent infection from blood transfusion in areas of high incidence and prevalence of HIV all but absolutely essential transfusions should be avoided, and donors with low incidence of HIV infection should be selected.


Higher eucaryotic cdc25 proteins are structurally related to phosphoseryl/threonyl protein phosphatases. cdc25 proteins are universally involved in the control of cell division. Using an original method of sequence analysis, cdc25 proteins from different sources were compared to protein phosphatases. Protein phosphatases could clearly be characterized as two distinct protein families, the phospho-seryl/threonyl phosphatases, and the phospho-tyrosyl phosphatases. None of the cdc25 proteins analyzed fitted with the phospho-tyrosyl phosphatases, indicating that if they indeed possess this biochemical activity, they form a distinct phsophatase protein group. Unexpectedly, higher eucaryotic cdc25 proteins (from human and fly) were found to be structurally related to phospho-seryl/threonyl phosphatases. These results fit well with expected function of the proteins, associated solely in higher eucaryotes, to dephosphorylation of threonine in the cell cycle control protein cdc2.


Herpes simplex virus and cytomegalovirus in pregnant Chilean university students. Virus isolation tests were performed during the first, second, and third trimesters of pregnancy on a group of 163 students who received prenatal care at the University of Chile's Student Medical and Dental Service to determine the possible existence of genital herpes or cytomegalovirus infections. Vaginal secretion samples tested for herpes simplex virus (HSV) and urine samples tested for cytomegalovirus (CMV) were inoculated onto cell cultures and examined by immunofluorescent procedures using monoclonal antibodies. HSV was detected in five subjects, only one of whom had herpetic lesions, and was identified as type 2 in all cases. Cytomegalovirus was detected in seven subjects.


Cannabinoid receptor agonists inhibit Ca current in NG108-15 neuroblastoma cells via a pertussis toxin-sensitive mechanism. Cannabinoid receptor ligands irreversibly inhibited peak voltage-activated Ca currents (44%) in NG108-15 cells; this inhibition was Pertussis toxin-sensitive. Inhibition was largely due to a reduction in the omega-conotoxin sensitive portion of high-voltage activated (HVA) current, although there was also a significant decrease in low-voltage activated current (56%) and in the nifedipine-sensitive portion of HVA current (41%).


Vagal afferent modulation of nociception. Chemical, electrical or physiological activation of cardiopulmonary vagal (cervical, thoracic or cardiac), diaphragmatic vagal (DVAG) or subdiaphragmatic vagal (SDVAG) afferents can result in either facilitation or inhibition of nociception in some species. In the rat, these effects depend upon vagal afferent input to the NTS and subsequent CNS relays, primarily in the NRM and ventral LC/SC, although specific relay nuclei vary as a function of the vagal challenge stimulus. Spinal pathways and neurotransmitters have been identified for vagally mediated effects on nociception and consistently implicate the involvement of descending 5-HT and noradrenergic systems, as well as intrinsic spinal opioid receptors. Species differences may exist with respect to both the effects of DVAG and SDVAG afferents on nociception and the efficacy of vagal afferents to modulate nociception. However, it is also possible that such differences reflect the modality of noxious input (e.g., visceral versus cutaneous), the type of neuronal activity investigated (e.g., resting versus noxious-evoked), spinal location of recording (e.g., thoracic versus lumbosacral) and/or parameters of stimulation. It is also possible that activation of some vagal afferents is aversive, but whether this contributes to changes in nociception produced by vagal activation has not clearly been established. Finally, the vagal-nociceptive networks described in this review provide a fertile area for future study. These networks can provide an understanding of physiological and pathophysiological peripheral events that affect nociception.


Biological activity and metabolism of 20-hydroxyeicosatetraenoic acid in the human platelet. 1. The cytochrome P-450 metabolite of arachidonic acid, 20-hydroxyeicosatetraenoic acid (20-HETE), was found to be a potent, dose-dependent inhibitor of platelet aggregation and inhibitor of thromboxane biosynthesis induced by arachidonic acid (IC50 5.2 +/- 1.5 microM), A23187 (IC50 16.2 +/- 5.4 microM), and U46619 (IC50 7.8 +/- 2.4 microM). 20-HETE did not inhibit thrombin-induced aggregation. 2. The human platelet metabolized 20-HETE to a series of novel metabolites formed by cyclo-oxygenase as well as lipoxygenase pathways. The structures of the metabolites were identified by mass spectrometry as 20-hydroxy-thromboxane B2, 12,17-dihydroxyheptadecatrienoic acid, 12,20-dihydroxyeicosatetraenoic acid, and 11,20-dihydroxyeicosatetraenoic acid. 3. The identification of the 11-hydroxy metabolite of 20-HETE suggests that 20-HETE is less efficiently cyclized to an endoperoxide intermediate by cyclo-oxygenase than is arachidonate. 4. Although some biological activity of 20-HETE may be related to competition with endogenous arachidonate for cyclo-oxygenase metabolism, the predominant mechanism of action of 20-HETE appears to be through antagonism of the prostaglandin H2/thromboxane A2 receptor.


Experience in the surgical management of the hand in dystrophic epidermolysis bullosa. Between 1981 and 1990, 45 patients (80 hands) underwent 122 hand operations, involving extensive release of contractures and pseudosyndactyly, with split skin grafting of secondary defects. Other procedures included the use of Kirschner wires, tendon release and arthroplasty. A postoperative programme of long term night splintage was employed. Finger extension was significantly improved for up to 5 years postoperatively, with a mean extension deficit preoperatively of 139 degrees, at 1 year postoperatively 60 degrees (p < 0.0001) and 4 years 84 degrees (p < 0.05). Preoperatively, pseudosyndactyly was present in 64.3% of web spaces, in comparison to 13.3% at 1 year, 37% at 4 years (p < 0.01) and 66.6% at 5 years (N.S.) postoperatively. Adduction contracture of the thumb was significantly improved for 2 years postoperatively. Functional assessment of 18 hands showed significant improvement postoperatively.


The contribution of alpha-adrenoceptors to neurally-mediated contractions of the rabbit urethral smooth muscle. 1. The nature of the nerve-mediated contractions in the urethral smooth muscle from the rabbit was studied in vitro. Field stimulation caused smaller contractile responses than in the detrusor of the rabbit. 2. There was no significant difference in response to field stimulation or exogenous agents acting on adrenoceptors between longitudinal and circular strips from the rabbit urethra. Histological studies showed that the urethral muscle is arranged in three layers, which run circularly and longitudinally. 3. Atropine had very little effect on the response to field stimulation, phentolamine almost abolished the contractile response to nerve stimulation and sometimes unmasked a relaxation. 4. The alpha 1-adrenoceptor blocking agent, prazosin, blocked both the contractile response to the alpha 1-receptor agonist phenylephrine and that to intrinsic nerve stimulation, with similar potencies. The alpha 2-blocking agent yohimbine shifted the dose-response curve of the contractile response to the alpha 2-agonist, clonidine, in a dose-dependent manner, 10(-7) M causing a 10 fold shift. This concentration had no effect on the response to intrinsic nerve stimulation, suggesting that alpha 2-receptors are not involved in the response. Higher concentrations of yohimbine caused a suppression of the nerve-evoked response which is assumed to be non-specific. 5. Noradrenaline, phenylephrine, and clonidine caused dose-dependent contractile responses in the rabbit urethral strips. The contractions induced by clonidine developed more slowly than those induced by noradrenaline and phenylephrine. 6. These results demonstrate that the rabbit urethral smooth muscle contains both alpha 1- and alpha 2-adrenoceptors, and the nerve-mediated contraction of the rabbit urethra is adrenergic in nature and mediated mainly via alpha 1-adrenoceptors.


Impaired cyclic nucleotide-mediated vasorelaxation may contribute to closure of the human umbilical artery after birth. 1. The mechanical and biochemical effects of agents that relax vascular smooth muscle either through elevation of guanosine 3':5'-cyclic monophosphate (cyclic GMP) or adenosine 3':5'-cyclic monophosphate (cyclic AMP) levels were compared in isolated ring preparations of human umbilical artery and rat aorta. Tone was established by preconstriction with 5-hydroxytryptamine. 2. The endothelium-dependent vasodilator calcium ionophore (A23187) (which stimulates endothelium-derived relaxing factor [EDRF] release and thus acts through soluble guanylyl cyclase), sodium nitroprusside (which stimulates soluble guanylyl cyclase directly), and atrial natriuretic peptide (which stimulates particulate guanylyl cyclase) relaxed rat aorta but not human umbilical artery. 3. Sodium nitroprusside, 10 microM, increased cyclic GMP levels from 10 to 390 pmol mg-1 protein at 2 min in rat aorta, as compared with a slower, relatively attenuated rise from 5 to 116 pmol mg-1 protein after 15 min in human umbilical artery. The rise in cyclic GMP in the umbilical artery was not significantly augmented by the cyclic GMP phosphodiesterase inhibitor, MB22948. Atrial natriuretic peptide increased cyclic GMP levels in rat aorta but not in human umbilical artery. 4. Forskolin, 10 microM, which stimulates both soluble and particulate adenylyl cyclase, maximally relaxed rat aorta and increased cyclic AMP levels from 15 to 379 pmol mg-1 protein at 15 min, but did not significantly relax or increase cyclic AMP levels in human umbilical artery. After preincubation with the cyclic nucleotide phosphodiesterase inhibitor, IBMX, 10 microM forskolin increased cyclic AMP levels to 1365 pmol mg-1 protein at 30 min in human umbilical arteries, but these high levels were not accompanied by mechanical relaxation.5. 8-Bromo-cyclic GMP and 8-bromo-cyclic AMP which are lipophilic analogues of cyclic GMP and cyclic AMP, both maximally relaxed the rat aorta at a concentration of 10 microM, but did not significantly relax the human umbilical artery.6. The findings indicate that elevated cyclic nucleotide levels are not associated with mechanical relaxation of the post-partum human umbilical artery, as in other vessels such as rat aorta. This impaired response to cyclic nucleotides may contribute to closure of the umbilical artery after birth.


Cibacron blue stimulation of surfactant secretion in rat type II pneumocytes. 1. The effect of cibacron blue, a selective P2y-purinoceptor antagonist in some systems, on the stimulatory effect of adenosine 5'-triphosphate (ATP) on [3H]-phosphatidylcholine secretion was examined in primary cultures of rat type II pneumocytes prelabelled by overnight culture with [3H]-choline. 2. Cibacron blue alone stimulated phosphatidylcholine secretion in a concentration-dependent manner in the range 10(-4)-10(-3) M. At a concentration of 10(-4) or lower, cibacron blue had no effect on ATP-induced phosphatidylcholine secretion but at 10(-4)-10(-3) M it increased the effect of ATP. Enhancement of the ATP effect was apparent whether cibacron blue was added before or together with ATP. Cibacron blue also increased ATP-induced secretion in the presence of the P1-purinoceptor antagonist, xanthine amine congener (10(-5) M). 3. The stimulatory effect of cibacron blue on phosphatidylcholine secretion was additive to those of 5' (N-ethylcarboxyamido) adenosine (NECA) and terbutaline but less than additive to that of ATP. 4. Cibacron blue alone had no effect on formation of cyclic AMP or inositol phosphate and when added simultaneously with ATP it did not affect the ATP-induced increase in these second messengers. Preincubation of the cells with cibacron blue before addition of ATP, however, resulted in antagonism of the ATP-induced increase in cyclic AMP and inositol phosphates. Preincubation with ATP had the same effect. The stimulatory effects of NECA and terbutaline on cyclic AMP formation were enhanced by preincubation with cibacron blue. 5. Thus, ATP-induced phosphatidylcholine secretion in type II cells is not diminished by the P2y-antagonist, cibacron blue.5. Thus, ATP-induced phosphatidylcholine secretion in type II cells is not diminished by the P2yantagonist, cibacron blue. On the contrary, cibacron blue stimulates phosphatidylcholine secretion. Cibacron blue may act as a P2-agonist in type II pneumocytes.


Effects of Hoe 140, a bradykinin B2-receptor antagonist, on renal function in conscious normotensive rats. 1. The present study was designed to determine if endogenous kinins are involved in the regulation of arterial blood pressure and renal function in conscious rats given deoxycorticosterone enantate (DOC, 25 mg kg-1, s.c., weekly) or vehicle for two weeks. 2. The bradykinin B2-receptor antagonist, D-Arg[Hyp3,Thi5,D-Tic7,Oic8]- bradykinin (Hoe 140), at a dose of 300 micrograms kg-1, s.c., blocked the hypotensive effect of 300 ng kg-1 bradykinin i.a., but it did not alter the blood pressure lowering action of 300 ng kg-1 acetylcholine or prostaglandin E2. Inhibition of the response to bradykinin persisted up to 6 h after the administration of Hoe 140. 3. Administration of 300 micrograms kg-1 Hoe 140 s.c. four times a day did not alter mean blood pressure, renal blood flow, or renal function in rats given DOC-vehicle. However, it decreased urinary volume by 70% (from 48.2 +/- 3.8 to 14.3 +/- 3.7 ml 24 h-1, P less than 0.01) and urinary secretion of sodium by 54% (from 1.02 +/- 0.05 to 0.47 +/- 0.16 mmol 24 h-1, P less than 0.01) and potassium by 30% (from 2.93 +/- 0.15 to 2.04 +/- 0.15 mmol 24 h-1, P less than 0.05) in DOC-treated rats. Mean blood pressure, glomerular filtration rate and total renal blood flow remained unchanged. 4. Our results suggest that endogenous kinins play a role in the regulation of renal excretion of water and sodium in the presence of elevated levels of DOC.


Multiple actions of glaucine on cyclic nucleotide phosphodiesterases, alpha 1-adrenoceptor and benzothiazepine binding site at the calcium channel. 1. In the present study, the properties of glaucine (an aporphine structurally related to papaverine) were compared with those of papaverine, diltiazem, nifedipine and prazosin. The work includes functional studies on rat isolated aorta contracted with noradrenaline, caffeine or KCl, and a determination of the affinity of glaucine at calcium channel binding sites of alpha-adrenoceptors, by use of [3H]-(+)-cis-diltiazem, [3H]-nitrendipine and [3H]-prazosin binding to cerebral cortical membranes. The effects of glaucine on the different molecular forms of cyclic nucleotide phosphodiesterases (PDE) isolated from bovine aorta were also determined. 2. Contraction evoked by noradrenaline (1 microM) or depolarizing solution (60 mM KCl) were inhibited in a concentration-dependent manner by all the compounds tested. As expected, prazosin showed a greater selectivity of action on NA-induced contraction, whereas nifedipine and diltiazem appeared more potent on KCl-induced contraction. Glaucine had a greater potency on the contraction elicited by noradrenaline whereas papaverine acted non specifically. 3. In Ca(2+)-free solution, prazosin (0.1 microM) and glaucine (0.1 mM) inhibited the contraction evoked by NA; diltiazem (0.1 mM) diminished this contraction whereas nifedipine (1 microM) had no effect. Preincubation of tissues with glaucine, diltiazem, nifedipine and prazosin did not modify the contractile response induced by caffeine. In contrast, papaverine (0.1 mM) significantly inhibited the contractions evoked by NA or caffeine in Ca(2+)-free medium. 4. Glaucine and papaverine show affinity at the [3H]-prazosin binding site and at the benzothiazepine binding site of the Ca(2+)-channel receptor complex, but have no effect at the dihydropyridine binding site in rat cerebral cortex. Glaucine exerts some selectivity as an inhibitor of [3H]-prazosin binding as opposed to [3H]-(+ )-cis-diltiazem binding while papaverine appears to have approximately equal affinity in this respect.5. This study confirms the presence of four phosphodiesterase (PDE) activities in bovine aorta: a calmodulin-activated PDE (CaM-PDE type I) which hydrolyzed preferentially guanosine 3':5'-cyclic monophosphate (cyclic GMP); a cyclic GMP selective form (cGMP-PDE type V); and two low Km adenosine 3':5'-cyclic monophosphate (cyclic AMP) PDEs that are insensitive to the stimulatory effect of CaM, one of which was inhibited by cyclic GMP (CGI-PDE, type III) and the other by rolipram (cAMP-PDE, type IV). Glaucine selectively inhibits one of the two forms of Ca2+-independent low Km cAMP-PDE, the type IV. In contrast, papaverine exerts a non-selective inhibitory effect upon all PDE forms.6. The present work provides evidence that glaucine, a benzyltetrahydroisoquinoline alkaloid, has interesting properties as an alpha l-adrenoceptor antagonist, calcium entry blocker (through the benzothiazepine recognition site in the calcium channel) and as a selective inhibitor of the rolipram-sensitive cAMP-PDE, type IV PDE.


Inhibitory effect of methysergide on calcitonin gene-related peptide-induced vasodilatation and ocular irritative changes in the rabbit. 1. Calcitonin gene-related peptide (CGRP) is involved in ocular neurogenic inflammation in the rabbit, causing vasodilatation in the anterior uvea, breakdown of the blood-aqueous barrier, increase in the intraocular pressure (IOP) and rise in the adenosine 3':5'-cyclic monophosphate (cyclic AMP) content in the aqueous humour. So far there is no means of preventing these CGRP-induced ocular effects. 2. In the present study, the effect of intravenous methysergide (1-10 mg kg-1, b.w.) on CGRP-induced changes in the IOP, blood-aqueous barrier and cyclic AMP content in the aqueous humour was studied in vivo. The effect of methysergide on CGRP-induced vasodilatation both in vivo and in vitro was also investigated. 3. Methysergide decreased intraocular pressure but had only a transient effect on blood pressure. Methysergide decreased the regional blood flow in ocular tissues by 53-65%, but did not have such a vasoconstrictor effect in most extra-ocular tissues studied. 4. Methysergide inhibited CGRP-induced vasodilatation, increase in the IOP, breakdown of the blood-aqueous barrier and increase in the cyclic AMP content in the aqueous humour in vivo. 5. In vitro, methysergide alone did not have effects on the vascular tone in isolated ophthalmic artery of rabbit. However, it potentiated noradrenaline (NA)-induced contraction. There were no differences in the IC50 values for CGRP on the NA-induced contraction in the presence and absence of methysergide, indicating that methysergide has no direct effect on the vasorelaxant effect of CGRP in vitro. 6. The present study demonstrates that in the rabbit eye methysergide inhibits CGRP-induced changes.One inhibitory mechanism of methysergide may be to enhance the effect of a vasoconstrictor (NA) to antagonize the vasodilator effect of CGRP. The present findings suggest that a methysergide-sensitive mechanism may be used to limit some pathophysiological conditions in the eye that involve neurogenic inflammation and the release of CGRP.


CP-93,129, sumatriptan, dihydroergotamine block c-fos expression within rat trigeminal nucleus caudalis caused by chemical stimulation of the meninges. 1. The effects of intravenously administered 5-HT1B receptor agonists were examined on c-fos like immunoreactivity, an indicator of neuronal activation, within the brain stem. C-fos was induced by injecting an algesic, vasoconstrictor substance (0.3 ml of autologous blood) or a pro-inflammatory molecule, carrageenin (1 mg in 0.1 ml saline) into the cisterna magna of pentobarbitone-anaesthetized Sprague-Dawley rats and was visualized in serial sections (50 micrometers) by use of a polyclonal antiserum. 2. As previously reported, the injection of blood caused significant labelling within laminae I, IIo of the trigeminal nucleus caudalis, a major nociceptive brain stem nucleus, as well as within nucleus of the solitary tract and area postrema. A similar pattern of expression with fewer cells per section was detected after carrageenin instillation. The number of expressing cells was reduced by 54% in trigeminal nucleus caudalis but not within the nucleus of the solitary tract or area postrema when blood was injected in adult rats neonatal capsaicin treatment. 3. Pretreatment with 5-HT1 agonists with some selectivity for the 5-HT1B receptor, CP-93,129 (460 nmol kg-1 x 2, i.v.), sumatriptan (720 nmol kg-1 x 2, i.v.) or dihydroergotamine (86 nmol kg-1 x 2, i.v.) reduced positive cells by 39%, 31%, and 33% respectively in trigeminal nucleus caudalis but not in nucleus of the solitary tract or area postrema after blood instillation. Pretreatment with the analgesic morphine (15 mumol kg-1, s.c.) also decreased the number of positive cells by 63% in trigeminal nucleus caudalis. 4. CP-93,129 (460 nmol kg-1 x 2, i.v.) reduced the number of c-fos labelled cells by 47% within lamina I, IIo after carrageenin instillation. 5. Drug-induced blockade appeared to be tissue-dependent. Pretreatment with sumatriptan (720 nmol kg-1 x 2, i.v.) did not block c-fos expression in trigeminal nucleus caudalis following formalin application to the nasal mucosa.6. Drug-induced blockade may be mediated by an action on primary afferent (trigeminovascular) fibres in as much as CP-93,129 (460 nmol kg-' x 2, i.v.) did not reduce the number of expressing cells within the trigeminal nucleus caudalis following blood instillation in rats treated as neonates with capsaicin.7. We infer from these results that the analgesic actions of agonists at 5-HTB receptors (the receptor subtype analogous to 5-HTID in man) need not depend upon the presence of vasodilatation and, that 5-HTID receptor-mediated blockade of neurotransmission contributes significantly to the analgesic effects of these drugs in headache.8. Based on the demonstrated effects of 5-HTB/D agonists against the actions of two chemicallyunrelated meningeal stimulants, we suggest that treatment with 5-HTID agonists may be useful for the alleviation of pain in other headache conditions associated with meningeal irritation. Bacterial, viral(including AIDS meningovascular inflammation) and other forms of chemical meningitis merit further investigation.


Peripheral analgesic activities of peptides related to alpha-melanocyte stimulating hormone and interleukin-1 beta 193-195. 1. The hyperalgesic effects of interleukin-1 beta (IL-1 beta) and prostaglandin E2 (PGE2) were measured in rats. 2. Hyperalgesic responses to IL-1 beta were inhibited in a dose-dependent manner by alpha-melanocyte stimulating hormone (alpha-MSH)-related peptides with the following order of potency: [N1(4),D-Phe7]alpha-MSH greater than alpha-MSH greater than Lys-D-Pro-Val greater than Lys-Pro-Val greater than Lys-D-Pro-Thr greater than D-Lys-Pro-Thr. 3. Hyperalgesic responses to PGE2 were not inhibited by Lys-D-Pro-Thr and D-Lys-Pro-Thr but were inhibited in a dose-dependent manner by the other peptides with the same order of potency as against IL-1 beta. 4. The potencies of [N1(4), D-Phe7]alpha-MSH and alpha-MSH were greatly diminished by deletion of their C-terminal tripeptide, Lys11-Pro-Val13. 5. Nor-binaltorphimine (Nor-BNI) largely reversed the analgesic effects of alpha-MSH, [N1(4), D-Phe7]alpha-MSH, Lys-Pro-Val and Lys-D-Pro-Val indicating that kappa-opioid receptors mediated the analgesic activity of these peptides. 6. Nor-BNI did not antagonize the inhibition by Lys-D-Pro-Thr and D-Lys-Pro-Thr of IL-1 beta evoked hyperalgesia indicating that these peptides were not acting via kappa-opioid receptors.


[3H]-RS-15385-197, a selective and high affinity radioligand for alpha 2-adrenoceptors: implications for receptor classification. 1. RS-15385-197 is the most potent and selective alpha 2-adrenoceptor antagonist available. We have used [3H]-RS-15385-197 to define alpha 2-adrenoceptor subtypes. The binding of [3H]-RS-15385-197 to membranes of rat cerebral cortex, rat neonatal lung and human platelets was reversible, saturable and of high affinity. Saturation experiments indicated that [3H]-RS-15385-197 bound to a single population of sites in all 3 tissues with high affinity (0.08-0.14 nM). The density of sites labelled by [3H]-RS-15385-197 was greater in the cortex (275 fmol mg-1 protein) than in the neonate lung (174 fmol mg-1 protein) and human platelet (170 fmol mg-1 protein). The density of sites labelled with [3H]-RS-15385-197 in the cortex was significantly greater than that labelled with [3H]-yohimbine (121 fmol mg-1 protein). 2. The selective alpha 2-adrenoceptor antagonists, idazoxan, yohimbine, rauwolscine and WY 26703 displaced [3H]-RS-15385-197 binding to rat cerebral cortex in a simple manner with Hill slopes close to unity. The affinities derived for these antagonists against [3H]-RS-15385-197 were similar to the values obtained for the displacement of [3H]-yohimbine indicating the alpha 2-adrenoceptor nature of the binding site. 3. alpha 2A-Adrenoceptor selective compounds, oxymetazoline and BRL 44409, showed high affinity for [3H]-RS-15385-197 binding in the human platelet and lower affinity in the neonate lung, while the alpha 2B-selective compounds, prazosin and imiloxan, showed high affinity for [3H]-RS-15385-197 binding in the neonate lung.This suggests that [3H]-RS-15385-197 labels both alpha2A- and alpha2B-adrenoceptor subtypes.4. Prazosin and methysergide inhibited the binding of [3H]-RS-15385-197 in the rat cerebral cortex in a simple manner consistent with an interaction at a single site. Although oxymetazoline inhibited [H]-RS- 15385-197 with a Hill slope significantly different from unity, the slope was increased to unity in the presence of Gpp(NH)p, suggesting an agonist-like interaction.5. The site labelled by [3H]-RS-15385-197 in the rat cortex shows high affinity for oxymetazoline and low affinity for prazosin which could be taken as evidence for classifying the site as an alpha2A-subtype.However, the affinities of yohimbine, rauwolscine and oxymetazoline at this site do not correspond to the population of sites in the human platelet. Yohimbine and rauwolscine were 20 fold selective for the platelet alph2A-subtype, whereas phentolamine was 2 fold and imiloxan was 10 fold selective for the cortex subtype. Indeed although the site showed some similarities with the alpha2A-subtype, the highest degree of homology was observed between this site and the rat submaxillary gland and the RG20 clone,tentatively called the alpha2D-adrenoceptor subtype. We propose that the alpha2-adrenoceptor in the rat cortex may therefore correspond to the putative alpha2D-subtype of the adrenoceptor.


ATP mediates excitatory synaptic transmission in mammalian neurones. Adenosine 5'-triphosphate (ATP, 0.1-100 microM), produced inward currents in patch-clamped coeliac neurones from guinea-pig when studied in either the whole cell configuration or in excised (outside-out) patches. The P2-purinoceptor antagonists suramin (80-230 microM) or reactive blue 2 (2-20 microM) depressed the ATP-induced currents but not those produced by acetylcholine. Excitatory post-synaptic currents (e.p.s.cs) were observed in cultured neurones. E.p.s.cs had similar current-voltage relationships to currents evoked by ATP in excised patches and were reduced by suramin or reactive blue 2 to a similar extent as ATP currents. The results suggest that ATP is the excitatory neurotransmitter in cultures of these neurones.


Adenosine receptors on human airway epithelia and their relationship to chloride secretion. 1. We have characterized an adenosine receptor subtype present in human airway epithelial cells by measuring the changes in the intracellular levels of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and the rate of transepithelial Cl- secretion. 2. Primary cultures of human nasal epithelium obtained from excised surgical airway epithelial tissues and the cell lines BEAS39 and CF/T43 derived from human airway epithelium were grown on plastic dishes and labelled with [3H]-adenine for measurement of intracellular cyclic AMP accumulation. Primary cultures were loaded with the calcium indicator fura-2 to measure [Ca2+]i and studied as polarized, ion transporting epithelia on collagen matrix supports for measurement of Cl- secretion. 3. Adenosine analogues stimulated cyclic AMP accumulation with a rank order of potency characteristic of an A2-receptor: 5-N-ethyl-carboxamidoadenosine (NECA) greater than adenosine greater than R-phenylisopropyladenosine (R-PIA), 6-N-cyclopentyladenosine (CPA) greater than S-PIA. NECA increased cyclic AMP accumulation in normal and cystic fibrosis (CF) primary cells as well as in the CF/T43 and BEAS39 cell lines with K0.5 values ranging from 0.3 to 3 microM. Preincubation with NECA resulted in the homologous desensitization of airway epithelial cells. The effect of NECA was specifically inhibited by the adenosine receptor antagonist, aminophylline, in a competitive manner. 4. The A1-adenosine receptor agonists CPA and R-PIA did not inhibit isoprenaline-stimulated cyclic AMP accumulation in CF/T43 cells, and potentiating effects of the adenosine analogues were observed on forskolin-stimulated cyclic AMP accumulation. Adenosine analogues did not cause significant changes in intracellular Ca2+ ([Ca2+]i) in airway epithelium.5. Adenosine analogues, applied to either the serosal or mucosal side of the polarized amiloride pretreated primary cultures, induced changes in I,, with a rank order of potency of agonists similar to that observed for stimulation of cyclic AMP accumulation. Intracellular microelectrode studies indicated that the locus of action was the apical membrane Cl- conductance. Adenosine failed to stimulate C1- secretion in CF airway epithelium.6. These results provide evidence for the existence of an A2-adenosine receptor that modulates intracellular levels of cyclic AMP in human airway epithelium. Activation of this receptor might lead to stimulation of Cl- secretion in amiloride pretreated normal but not CF cells.


GR94839, a kappa-opioid agonist with limited access to the central nervous system, has antinociceptive activity. 1. The pharmacological profile of GR94839, a kappa-opioid agonist with limited access to the central nervous system, has been investigated. Its antinociceptive activity has been compared with that of GR103545, a centrally-penetrating kappa-agonist and ICI204448, the previously described peripherally-selective kappa-agonist. 2. GR94839 was a potent agonist in the rabbit vas deferens in vitro assay for kappa-opioid receptors (IC50: 1.4 +/- 0.3 nM; n = 6), but had limited activity at mu- or delta-opioid receptors. 3. In the mouse abdominal constriction test, GR94839 was 238 fold more potent when given i.c.v. (ED50: 0.008 (0.004-0.029) mg kg-1; n = 18) than when s.c. (ED50: 1.9 (0.7-3.1) mg kg-1; n = 30). In comparison, GR103545 was equipotent when given i.c.v. or s.c. 4. After intravenous administration, the maximum plasma to brain concentration-ratio attained by GR94839 was 18 compared with 2 for GR85571, a structurally-related kappa-agonist that is centrally-penetrating. 5. GR94839 inhibited the 2nd phase of the rat formalin response at doses 7 fold lower than those required to inhibit the 1st phase (ED50 vs 1st phase: 10.2 (6.7-17.1) mg kg-1, s.c.; ED50 vs 2nd phase: 1.4 (1.0-1.8) mg kg-1, s.c.; n = 18). GR103545 was equipotent against the two phases. 6. Intraplantar administration of the opioid antagonists, norbinaltorphimine (100 micrograms) or naltrexone (1 microgram), reversed the antinociceptive effect of systemic GR94839 (3 mg kg-1, s.c.) against the 2nd phase of the formalin response and intraplantar injection of GR94839 (30-100 micrograms) selectively inhibited the 2nd phase.7. GR94839 and IC1204448 reversed the hyperalgesia in the zymosan-inflamed rat paw at doses (ED50 GR94839: 2.0 (1.1-3.2) mg kg-', s.c.; ED50 IC1204448: 1.2 (0.8-1.7) mg kg-', s.c.), lower than those required to raise the noxious pressure threshold in the non-inflamed paw (EDSO GR94839: 16.4 (8.6-46.7) mg kg', s.c.; ED50 IC1204448: 68.0 (22.1-32000) mg kg', s.c.). GR103545 raised the noxious presure threshold in the inflamed and non-inflamed paws at the same doses.8. GR94839 was sedative in the rat rotarod test (ED50: 35 (12-245) mg kg-', s.c.) at doses higher than those required to inhibit the 2nd phase of the formalin response or reverse hyperalgesia in the zymosan-inflamed rat paw. The doses were comparable to those that inhibited the 1st phase of the formalin response and raised the noxious pressure threshold in the non-inflamed paw.9. The results suggest that GR94839 is a selective kappa-agonist which has antinociceptive activity against inflammatory pain at doses that produce limited central effects. These antinociceptive effects are probably mediated at peripheral opioid receptors.


Modulation of the pharmacological actions of nitrovasodilators by methylene blue and pyocyanin. 1. In superfused precontracted strips of rabbit aorta, methylene blue (MeB) or pyocyanin (Pyo, 1-hydroxy-5-methyl phenazinum betaine) at concentrations of 1-10 microM inhibited relaxations induced by endothelium-derived relaxing factor (EDRF), glyceryl trinitrate (GTN), S-nitroso-N-acetyl-penicillamine (SNAP) or 3-morpholino-sydnonimine (SIN-1). However, the vasorelaxant actions of sodium nitroprusside (NaNP) or sodium nitrite (NaNO2) were enhanced by MeB or Pyo. Oxyhaemoglobin (HbO2, 1 microM) inhibited the activities of EDRF and all of the nitrovasodilators studied. Vascular preparations were not relaxed by Pyo unless pretreated with NaNP (0.05-10 microM). 2. In bathed, precontracted rings of rabbit aorta, Pyo (10 microM) produced a shift to the left of the cumulative concentration-response curve for NaNP (0.01-10 microM). The rise in guanosine-3':5'-cyclic monophosphate (cyclic GMP) content of aortic tissue was also enhanced. 3. The vasorelaxant potency of NaNP (30 microM) at pH 5-8 and at 37 degrees C remained unchanged over 2.5 h while a solution of SNAP (30 microM) progressively lost its biological activity over 60 min. The in vitro degradation of the biological activity of SNAP was accelerated by MeB (150 microM) or Pyo (150 microM), whereas the vasorelaxant potency NaNP (30 microM) was doubled when incubated with MeB or Pyo. 4. In human platelet-rich plasma, MeB or Pyo (0.3-3.0 microM) uncovered an anti-aggregatory action of subthreshold concentrations of NaNP (4-8 microM). This was abrogated by HbO2 (10 microM).5. We conclude that MeB or Pyo differ from HbO2 in their mode of interaction with nitrovasodilators.HbO2 scavenges nitric oxide that is released from all types of nitrovasodilators. MeB and Pyo exert a similar action towards organic nitrovasodilators (e.g. SNAP, SIN-1). However, the pharmacological actions of inorganic nitrovasodilators (e.g. NaNP or NaNO2) are potentiated by MeB and Pyo owing to facilitation of the intracellular release of nitric oxide from the inorganic nitrovasodilators.


The effects of noradrenaline and adenosine 5'-triphosphate on polyphosphoinositide and phosphatidylcholine hydrolysis in arterial smooth muscle. 1. The effects of noradrenaline and alpha,beta,methylene adenosine 5'-triphosphate (alpha,beta,methylene ATP) on polyphosphoinositide metabolism, phosphatidylcholine hydrolysis and contraction in rabbit saphenous arteries were investigated. The effect of noradrenaline upon polyphosphoinositide metabolism was also investigated in the rat tail artery. 2. Noradrenaline (10(-7)-10(-4) M) evoked a concentration-dependent increase in total inositol phosphate accumulation in the rat tail but not in the rabbit saphenous artery. Propranolol (3 x 10(-6) M) did not alter this result in the rabbit saphenous artery. In addition, alpha,beta,methylene ATP (10(-6) M) significantly increased total inositol phosphate accumulation in the rabbit saphenous artery, while potassium chloride (8 x 10(-2) M) was ineffective. 3. Phorbol 1,2-myristate 1,3-acetate (3 x 10(-8) M) enhanced noradrenaline (10(-2)-10(-4) M)-evoked contractions in rabbit saphenous artery. The contractile responses to potassium chloride (1- 16 x 10(-2) M) in tissues treated with 6-hydroxydopamine (5 x 10(-4) M), in vitro, were unaffected by these concentrations of the phorbol ester. 4. Noradrenaline (10(-6)-10(-4) M) evoked a concentration-dependent increase in the levels of choline and choline phosphate, but not in those of glycerophosphocholine, in the rabbit saphenous artery. Choline levels increased significantly over the first 15-30 s then declined to control levels within 2 min of addition of noradrenaline (10(-5) M). A smaller initial rise in choline phosphate levels (15-30 s) was followed by a larger secondary rise at 2-4 min.5. alpha, beta, methylene ATP (10-1_ 0-4 M) also evoked a concentration-dependent increase in the levels of both choline and choline phosphate, but not those of glycerophosphocholine, in the rabbit saphenous artery. alpha, beta, methylene ATP (10-4 M) significantly increased levels of both of these products within the first 15-30 s of addition of the drug; these levels reached a stable plateau 1 min after addition.6. The maximum accumulation of choline or choline phosphate evoked by either noradrenaline or alpha, beta, methylene ATP, acting alone or in combination, was not significantly different. No evidence of synergism between noradrenaline and alpha, beta, methylene ATP was observed.7. This study demonstrates that each of the co-transmitters in the rabbit saphenous artery, noradrenaline and adenosine 5'-triphosphate (ATP), promote phosphatidylcholine hydrolysis. Noradrenaline seems to rely on phosphatidylcholine hydrolysis to mediate its contractile effects, whilst ATP promotes both polyphosphoinositide and phosphatidylcholine metabolism suggesting that multiple signal-transduction mechanisms are involved in stimulus-contraction coupling in this artery.


Tachykinin NK1 receptor in the guinea-pig isolated proximal urethra: characterization by receptor selective agonists and antagonists. 1. The tachykinin receptor mediating contraction of the guinea-pig isolated proximal urethra has been characterized by use of receptor selective agonists and antagonists. All experiments were performed in the presence of peptidase inhibitors (bestatin, captopril and thiorphan, 1 microM each) in order to reduce peptide degradation. 2. The natural tachykinins, substance P and neurokinin A produced a concentration-dependent contraction of rings of the proximal urethra which approached the same maximum (about 50% of the response to 80 mM KCl). Substance P (EC50 155 nM) was slightly (3.6 times) more potent than neurokinin A (EC50 560 nM). 3. The tachykinin NK1 receptor selective agonist, [Sar9]substance P sulphone (EC50 62 nM), was slightly more potent than substance P and produced the same maximal response of natural tachykinins. The NK2 receptor selective agonist, [beta Ala8] neurokinin A(4-10), was active only at microM concentrations and its maximal effect did not exceed 20% of that to substance P or neurokinin A. The NK3 receptor selective agonist, senktide, was ineffective up to 30 microM. 4. The response to [Sar9]substance P sulphone was antagonized in a competitive manner by either (+/-)-CP 96,345 (pA2 7.75, slope - 1.10) or GR 82,334 (pA2 7.31, slope - 1.26), which are selective NK1 receptor antagonists, while it was unaffected (up to 10 microM) by MEN 10,376, a selective NK2 receptor antagonist. 5. The response to 10 microM [beta Ala8]neurokinin A (4-10) was abolished by either 0.2 microM (+/-)-CP 96,345 or 1 microM GR 82,334, suggesting the involvement of NK1 receptors.6. Electrical field stimulation (5 and 10 Hz, 0.25 ms, 100 V, trains of 5 s duration) produced tetrodotoxin-sensitive phasic contractions of the urethra which were abolished by atropine plus phentolamine (3 microM each). Capsaicin (1 microM) produced a small transient contraction of the urethra which was abolished by ( )-CP 96,345 (0.1 microM). ( )-CP 96,345 did not modify the response to electrical field stimulation.7. We conclude that tachykinin NK, receptors are the main if not the only mediators of the contractile response of guinea-pig proximal urethra to peptides of this family and that this preparation is useful for assessing the affinities of various ligands for the NK, receptor. Endogenous tachykinins released from peripheral endings of capsaicin-sensitive primary afferents produce urethral contraction by activating NK, receptors.


Blockade of Na+ current by promethazine in guinea-pig ventricular myocytes. 1. To elucidate the antiarrhythmic mechanism of promethazine, its effects on the fast Na+ current (INa) were examined in single guinea-pig ventricular myocytes by whole-cell voltage clamp methods. 2. Promethazine blocked INa with a KD of 42.6 microM and Hill's coefficient of 1.1 at a holding potential of -140 mV. 3. The INa blockade was enhanced at a less negative holding potential of -80 mV with a change of KD to 4.4 microM. Although 10 microM promethazine did not change the inactivation time constants of INa, it shifted the steady-state inactivation curve (h infinity curve) toward more negative potentials by 19.5 mV with the slope factor unaffected. 4. Double pulse experiments revealed that the development of blockade followed two-exponential functions having time constants of 7 and 220 ms at -20 mV. 5. Promethazine slowed the repriming of INa. This was associated with the development of slow phase having a time constant of 1160 +/- 59 ms. 6. Promethazine produced a profound use-dependent block when the cell was repeatedly stimulated with interpulse intervals shorter than 1 s. However, short pulses of 2 ms duration hardly produced such a use-dependent block. Hence, open channel blockade is considered to play a minor role in the promethazine action on INa. 7. These results suggest that promethazine blocks cardiac INa in a manner similar to class I antiarrhythmic drugs and that this effect may account for its antiarrhythmic action.


Contribution of cyclic GMP formation to KRN2391-induced relaxation in coronary artery of the pig. 1. In the present study, we investigated the relationship between relaxation and guanosine 3':5'-cyclic monophosphate (cyclic GMP) formation induced by KRN2391, compared with those induced by nicorandil and nitroglycerin, in the coronary artery of the pig. 2. KRN2391 (10(-8)-3 X 10(-5) M), nicorandil (10(-8)-3 X 10(-4) M) and nitroglycerin (10(-9)-10(-5) M) antagonized the contraction caused by 25 mM KCl in a concentration-dependent manner. 3. The concentration-relaxation curves for KRN2391, nicorandil and nitroglycerin shifted rightward in the presence of methylene blue (10(-5) M). 4. KRN2391 (10(-6) M), nicorandil (10(-4) M) and nitroglycerin (10(-6) M) induced an increased in cyclic GMP. 5. The magnitude of the shift of the concentration-relaxation curve caused by methylene blue and the increase in cyclic GMP with KRN2391 were lower than those with nicorandil and nitroglycerin. 6. The adenosine 3':5'-cyclic monophosphate (cyclic AMP) level was not increased by KRN2391 even at a concentration that produced full relaxation. 7. The present results suggest that KRN2391-induced relaxation in the coronary artery of the pig is partly due to the increase in cyclic GMP formation through the stimulation of guanylate cyclase.


MDL 27,531 selectively reverses strychnine-induced seizures in mice. 1. Strychnine-sensitive glycine receptors are primarily localized in the brainstem and spinal cord where they are the major mediators of postsynaptic inhibition. A compound which acts functionally like a glycine receptor agonist would be potentially useful as a pharmacological tool and as a therapeutic agent for treating disorders of glycinergic transmission. 2. MDL 27,531 (4-methyl-3-methylsulphonyl-5-phenyl-4H-1,2,4-triazole) blocked strychnine-induced tonic extensor seizures in mice following either intraperitoneal (ED50 = 12.8 mg kg-1; 30 min) or oral (ED50 = 7.3 mg kg-1; 30 min) administration. Time course studies revealed a maximal effect at 30-60 min, though significant activity was still seen after 24 h. 3. MDL 27,531 was selective in antagonizing strychnine seizures and little or no activity was seen against seizures produced by other chemical convulsants (bicuculline; quinolinic acid; mercaptopropionic acid); by electrical stimuli (maximal electroshock); or by sensory stimuli (audiogenic seizure susceptible mice). MDL 27,531 blocked pentylenetetrazol-induced seizures with an ED50 = 55 mg kg-1. This profile differed from those of the anticonvulsants diazepam, valproic acid, and diphenylhydantoin. 4. The antagonism of strychnine seizures by MDL 27,531 occurred at doses that did not produce signs of sedation (suppression of spontaneous motor activity), motor ataxia (disruption of rotarod performance), muscle relaxation (inhibition of morphine-induced Straub tail), or CNS depression (potentiation of hexobarbitone sleep time). MDL 27,531 had less side effect potential (as derived from ratios obtained from the above measures) relative to those of the known muscle relaxants diazepam and baclofen. 5. Although MDL 27,531 behaved functionally like a selective agonist at the strychnine-sensitive glycine receptor, the compound did not alter the in vitro binding of [3H]-strychnine to mice brainstem/spinal cord membranes at concentrations of up to 100 microM. In further in vitro binding assays, MDL 27,531 at concentrations of up to 100 microM, did not displace the binding of [3H]-muscimol, [3H]-flunitrazepam, or["S]-t-butylbicyclophosphorthionate to rat cortical membranes. These ligands bind to the 7y-aminobutyric acid (GABA), benzodiazepine, and picrotoxin-convulsant binding sites, respectively.6. MDL 27,531 (10-100mgkg-', i.p.) enhanced binding of the benzodiazepine antagonist [3H]-Ro15-1788 to mouse cerebral cortex in vivo without directly affecting GABA levels.7. Ro 15-1788 (16, 32 mg kg-') significantly blocked the MDL 27,531 antagonism of strychnineinduced seizures, though this antagonism was not competitive. The same doses of Ro 15-1788 produced parallel rightward shifts in the dose-response curves for diazepam inhibition of pentylenetetrazol-induced seizures, consistent with a competitive antagonism.8. Thus, MDL 27,531 acts functionally like an agonist at the strychnine-sensitive glycine receptor in its capacity to reverse selectively strychnine-induced seizures. Though the precise mechanism of action of MDL 27,531 is unknown, MDL 27,531 may act at an allosteric site on the strychnine-sensitive receptor which produces agonist-like activity.


Cultured astrocytoma cells generate a nitric oxide-like factor from endogenous L-arginine and glyceryl trinitrate: effect of E. coli lipopolysaccharide. 1. The inhibitory activity of astrocytoma cells (0.25-3 x 10(5)) treated with indomethacin (10 microM) on platelet aggregation was enhanced by incubating the cells with E. coli lipopolysaccharide (LPS, 0.5 micrograms ml-1) for 18 h. This effect was attenuated when cycloheximide (10 micrograms ml-1) was incubated together with LPS. The inhibition of platelet aggregation by cells treated with LPS was potentiated by superoxide dismutase (60 u ml-1) and ablated by oxyhaemoglobin (oxyHb, 10 microM) or NG-monomethyl-L-arginine (L-NMMA, 30-300 microM). The effects of L-NMMA were reversed by co-incubation with L-arginine (L-Arg, 100 microM) but not D-arginine (D-Arg, 100 microM). LPS also increased the levels of nitrite in the culture media and this increase was ablated by co-incubation with L-NMMA (300 microM) or cycloheximide (10 micrograms ml-1). 2. Astrocytoma cells (0.5 x 10(5)) treated with indomethacin (10 microM) enhanced the platelet inhibitory activity of glyceryl trinitrate (GTN, 11-352 microM) but not that of sodium nitroprusside (4 microM). Furthermore, when incubated with GTN (200 microM) a 4 fold increase in the levels of guanosine 3':5'-cyclic monophosphate (cyclic GMP) was observed. These effects were abrogated by co-incubation with oxyHb (10 microM) but not with L-NMMA (300 microM). Treatment of the cells with LPS (0.5 micrograms ml-1) for 18 h did not enhance their capacity to form NO from GTN. 3. Thus, in cultured astrocytoma cells, LPS enhances the formation of nitric oxide from endogenous L-arginine.In addition, these cells can metabolize GTN to nitric oxide but this process is not enhanced by LPS stimulation.


Combination treatment with noradrenalin and serotonin reuptake inhibitors in resistant depression. Eight cases of resistant recurrent depression were treated with a combination of nortriptyline and a new serotonin reuptake inhibitor, with or without concurrent lithium therapy. Significant improvement was seen in all patients where other drug regimes and ECT had been ineffective. No adverse reactions occurred in any of our patients, seven of whom were elderly. The combination treatment was more effective than individual therapies alone.


Influence of 5-fluorouracil and folinic acid on colonic healing: an experimental study in the rat. Male Wistar rats were subjected to colonic resection and randomized to one of four groups: control group (intraperitoneal NaCl, intravenous NaCl); 5-fluorouracil (5-FU) group (intraperitoneal 5-FU, intravenous NaCl); folinic acid group (intraperitoneal NaCl, intravenous folinic acid); and 5-FU-folinic acid group (intraperitoneal 5-FU, intravenous folinic acid). Treatment was started immediately after surgery and continued until the animals were killed at 3 or 7 days. Anastomotic complications (abscesses or dehiscence) occurred in four of 33 animals in the control group, 12 of 36 in the 5-FU group, one of 32 in the folinic acid group and nine of 36 in the 5-FU-folinic acid group. Anastomotic and skin breaking strength did not differ between groups on day 3 but by day 7 were significantly reduced in the 5-FU group. In rats given 5-FU-folinic acid, breaking strength was also reduced, but less so than in the 5-FU group. Breaking strength in animals receiving folinic acid was similar to that in the control group. In this model colonic healing was impaired after intraperitoneal 5-FU administration, but when folinic acid was added no further deterioration occurred.


Corticotropin-releasing factor antagonist reduces emotionality in socially defeated rats via direct neurotropic action. Introduction of a socially naive male rat into the home territory of a resident counterpart results in agonistic interactions, leading to the rapid social defeat of the intruder. Exposure to the aggressive resident produces a stress-response profile consisting of neuroendocrine activation and coping behaviors such as submission. The present studies examined the dependence of these adaptive responses on endogenous brain Corticotropin-Releasing Factor (CRF), a peptide hormone known to coordinate neuronally mediated- and pituitary-adrenal responses to stress. The Elevated Plus-Maze was employed as an animal model of emotionality in which stressors reduce subsequent exploration of open maze arms without walls in favor of enclosed maze arms. A CRF antagonist, alpha-hel CRF9-41, administered intracerebroventricularly (5 and 25 micrograms i.c.v.) immediately post-stress and 5 min prior to maze testing reversed the heightened emotionality produced by the resident exposure stressor. This action paralleled that of an anxiolytic dose of the short-acting benzodiazepine, midazolam (1.5 mg/kg i.p.). Intra-amygdaloid administration of lower doses of the CRF antagonist (125, 250 and 500 ng i.c.) also reversed, dose-dependently, the effect of exposure to an aggressive resident without altering the behavior of unstressed control animals. Further, the enhanced release of ACTH and corticosterone following social conflict was not modified over the short term by the intra-amygdaloid dose of CRF antagonist (250 ng i.c.) which was effective in reversing stress-induced hyper-emotionality. These results suggest that limbic system CRF substrates exert an anxiogenic effect on the exploratory behavior of socially defeated rats via a pituitary-adrenal-independent mechanism.


Lesioning of the nucleus basalis of Meynert has differential effects on mu, delta and kappa opioid receptor binding in rat brain: a quantitative autoradiographic study. Opioid receptor binding was investigated in rat brain following lesioning of the nucleus basalis of Meynert (nbM). The nbM, which provides cholinergic input to the cortex, was lesioned unilaterally using ibotenic acid. The efficacy of lesioning was confirmed by the observation of a significant decrease in choline acetyltransferase (ChAT) activity in the ipsilateral prefrontal cortex. The specific laminar and regional distribution of mu, delta and kappa opioid receptor binding was quantitated in various cortical and limbic structures in the rat using autoradiography. Distinct medial to lateral gradients of mu and kappa opioid binding were observed in regions of the cerebral cortex. In the lesioned hemisphere the levels of mu, delta and kappa opioid binding were altered in localized areas of the cerebral cortex and the hippocampus. The direction of these binding changes varied with the opioid receptor type being assessed. Delta opioid binding was increased in the lateral portions of the frontal, occipital, perirhinal and retrosplenial granular cortices. Kappa opioid binding was increased in the lateral portion of the occipital cortex and in the CA3 region of the hippocampus. In contrast, mu opioid binding was decreased in the lateral portions of the frontal, entorhinal and forelimb cortices. These opioid receptor changes are discussed with respect to the interactions between the cholinergic and opioid systems, and relevance of the nbM lesion model to Alzheimer's disease.


Mapping of prostaglandin E2 binding sites in rat brain using quantitative autoradiography. The density of specific prostaglandin E2 (PGE2) binding sites was quantitatively mapped in the rat brain using in vitro autoradiography. The anterior wall of the third ventricle and the nucleus solitary tract were found to have a very high density of binding sites (greater than 15 fmol/mg tissue). Two thalamic nuclei (paraventricular and anteroventral nuclei) and the dorsal parabrachial nucleus contained a high density of binding sites (10-15 fmol/mg tissue). Entorhinal cortex, ventral hippocampus, amygdala, dorsomedial hypothalamus, mammillary complex, some thalamic nuclei, central gray, superior colliculus, raphe nuclei, locus coeruleus, spinal trigeminal nucleus (caudal part) and the dorsal horn of the spinal cord (laminae 1 and 2) had each a moderate density of binding sites (5-10 fmol/mg tissue). Binding tended to occur in brain regions rich in neuronal cell bodies or neuronal cell processes (dendrites and axon terminals). PGE1, whose central actions are very similar to those of PGE2, had essentially the same pattern of binding sites as did PGE2 throughout the entire brain, suggesting there are receptors common to these two PGEs. In addition to already known functions of receptors common to these two PGEs. In addition to already known functions of PGE2 in the hypothalamus, which include fever genesis, promotion of wakefulness, cardiovascular control and LH-RH release, the unique distribution of extrahypothalamic PGE2 binding sites found in this study suggests its involvement in the processing or modulation of viscerosensory, somatosensory (nociceptive and possibly thermal) and visual inputs as well as in the central integration of autonomic and limbic functions.(ABSTRACT TRUNCATED AT 250 WORDS)


Naloxone benzoylhydrazone, a kappa 3 opioid agonist, stimulates food intake in rats. Naloxone benzoylhydrazone (NalBzoH) is a selective, short-acting agonist at the kappa 3 opioid receptor and a slowly dissociating potent antagonist at the mu opioid receptor. Given the important role of kappa receptors in the opioid control of food intake, the present study examined the central and peripheral effects of NalBzoH upon food intake. Central administration of NalBzoH (1-20 micrograms, i.c.v.) significantly increased food intake for up to 12 h, but failed to alter intake or body weight after 24 or 48 h. The 12 h duration of NalBzoH-mediated effects may be due to either persistent kappa 3 receptor occupancy, and/or activation of an ingestive system which maintains its activity. Peripheral administration of NalBzoH (20 mg/kg, s.c.) significantly increased food intake for up to 1 h. To distinguish kappa 1 (U50,488H) and kappa 3 (NalBzoH) hyperphagic effects, these agonist effects were compared following pretreatment with either naltrexone or the kappa 1 antagonist, nor-binaltorphamine (Nor-BNI). Whereas naltrexone significantly reduced both U50,488H and NalBzoH hyperphagia, Nor-BNI blocked U50,448H, but not NalBzoH hyperphagia. These data indicate a distinct role for the kappa 3 receptor in ingestive behavior separable from that of kappa 1 effects.


Chronic cerebrovascular insufficiency induces dementia-like deficits in aged rats. Young and aged rats were subjected to cerebrovascular insufficiency (CVI) for 3 and 9 weeks. At the end of each time period, local cerebral blood flow (lCBF), spatial memory function, 31P- and 1H-NMR spectroscopy and imaging of the brains were evaluated in vivo. Morphometric counts of CA1 hippocampal neuron damage and staining for glial fibrillary acidic protein (GFAP) were done post-mortem. Results show that after 3 weeks of CVI, cortical and hippocampal lCBF was significantly reduced in young and aged animals respectively. In addition, young and aged rats at 3 weeks following CVI showed spatial memory deficits in the Morris water maze and elevation of 31P-phosphomonoester as measured by non-invasive NMR spectroscopy. At the same time period, in vivo 1H-microimaging (MRI) of brains showed areas of high signal intensity (suggesting local edema) localized asymmetrically to the right hippocampal region in young and aged CVI rats. Morphometry of the hippocampal CA1 sector at post-mortem confirmed the in vivo MRI changes and demonstrated that a significant percentage of the CA1 pyramidal cells were damaged after CVI. Nine weeks after CVI, hippocampal CBF reductions, spatial memory impairment, spectroscopic-microimaging changes and CA1 sector cell damage continued to be observed in the aged animals but were resolved in the young rat brains. In addition, GFAP immunoreaction progressively increased in the hippocampus of aged rats subjected to CVI for 9 weeks. It is concluded that cognitive, metabolic and morphologic damage was significantly more severe and longer lasting in aged than young rat brain after chronic CVI. The deficits observed in this rat model appear to mimic the early pathology reported in Alzheimer's disease and suggest that the present model could provide fundamental clues relative to the etiology and possible management of this dementia.


Effects of repeated administration of desipramine or electroconvulsive shock on norepinephrine uptake sites measured by [3H]nisoxetine autoradiography. To determine if repeated administration of desipramine (DMI) or electroconvulsive shock (ECS) regulate uptake sites for norepinephrine (NE) in rat brain, the binding of [3H]nisoxetine ([3H]NIS) was measured using quantitative autoradiography. Groups of animals were given DMI intraperitoneally, either a single injection or repeated doses of 10 mg/kg once daily for 21 days and were killed 48 h after the last injection. Another group of rats received ECS daily for 12 days (150 mA, 300 ms, 60 Hz) and was killed 24 h after the last shock. Repeated administration of DMI caused statistically significant decreases (20-40%) in the binding of [3H]NIS in 8 out of 17 brain regions measured; these areas included the hippocampus, thalamus and the amygdala. Acute treatment with DMI had no effect on the binding of [3H]NIS in any of the regions analyzed except the centrolateral nucleus of the amygdala. By contrast, except for the paraventricular nucleus of the thalamus where ECS caused a modest (20%) increase in binding, no other brain region was affected by ECS. Thus it appears that repeated administration of DMI and chronic ECS treatment have different effects on the binding of [3H]NIS to uptake sites for NE in rat brain.


Structure-function relationships for analogues of L-2-amino-4-phosphonobutanoic acid on the quisqualic acid-sensitive AP4 receptor of the rat hippocampus. Hippocampal CA1 pyramidal cell neurons are sensitized to depolarization by L-2-amino-4-phosphonobutanoic acid (L-AP4) following exposure to L-quisqualic acid (QUIS). We have examined the interaction of 43 structural analogues of L-AP4 with both the 'induction' site and the QUIS-sensitive AP4 site in rat hippocampus. The synthesis of cis- and trans-4-phosphonoxy-L-proline, 3-(RS)-amino-5-phosphonopentanoic acid and 2(RS)-amino-5-phenyl-4(RS)-phosphonopentanoic acid (gamma-benzyl AP4) are described. None of the test compounds interact with the induction site; thus L-QUIS remains the only compound known to induce this effect. However, one compound (L-2-amino-3-(5-tetrazolyl)-propanoic acid (L-aspartate tetrazole) 'pre-blocked' and reversed the effects of QUIS. In addition, the potency of 16 analogues increased more than 4-fold following exposure of slices to L-QUIS. Among these, L-AP4, L-AP5, 2-amino-4-(methylphosphino)butanoic acid (AMPB), and E-1(RS)-amino-3(RS)-phosphonocyclopentanecarboxylic acid (E-cyclopentyl AP4) displayed IC50 values of less than 0.100 mM after QUIS. The results presented here suggest that the QUIS-sensitive AP4 site requires a spatial configuration of functional groups similar to that present in E-cyclopentyl AP4. The presence of a primary amino group and a phosphorus-containing group (either monoanionic or dianionic) appear to be required, however, a carboxyl group is not essential for interaction. The pharmacology of the QUIS-sensitive AP4 site suggests that it is distinct from other known binding sites for L-AP4 in the central nervous system (CNS).


Opiate infusion into nucleus accumbens: contrasting effects on motor activity and responding for conditioned reward. In the present experiments, we examined the roles of opiate receptor subtypes in the nucleus accumbens in spontaneous motor behavior and responding for conditioned reward. Locomotor activity was measured using photocell cages and reward-related responding was determined in the conditioned reinforcement (CR) paradigm. In the CR paradigm, food-deprived rats were trained to associate a compound stimulus with food reward, and were subsequently tested for responding on a lever that resulted in presentation of the compound stimulus alone. In all experiments, various opiate agonists were microinjected into the nucleus accumbens. In the activity studies, morphine sulfate (mixed mu and delta agonist; 0.025, 0.25 and 2.5 micrograms/0.5 microliter) caused an initial inhibition followed by a disinhibition of activity while [D-Ala2-Met5]-enkephalin (DALA) (mixed mu and delta agonist; 0.25, 2.5 and 5.0 micrograms/0.5 microliter) elicited an immediate potentiation of locomotor activity. The behavioral profile following [D-Ala2-N-Me-Phe4-Gly-ol5]-enkephalin (DAMGO; mu agonist; 0.01, 0.1 and 1.0 micrograms/0.5 microliter) was similar to morphine. In contrast, [D-Pen2,5]-enkephalin (DPEN; delta agonist; 0.02, 0.2 and 2.0 micrograms/0.5 microliter) induced an immediate, but relatively short-lasting activation. Both DALA and DPEN also dose-dependently enhanced rearing; rearing was not affected by the other treatments. U50,488H (kappa agonist; 0.0186, 0.186 and 1.86 micrograms/0.5 microliter) had no effect on any aspect of motor activity. In contrast to effects on motor activity, none of the opiate agonists significantly potentiated responding for CR, although morphine infusion did tend to increase responding somewhat. In contrast, D-amphetamine (20 micrograms/0.5 microliter) did potentiate responding, as previously reported. A distinction between the effects of opiates and psychostimulants on reinforced responding is hypothesized based on differential modulation of dopamine release. These experiments demonstrate a dissociation of the effects of opiates on locomotor activity and reward-related responding.


In vitro regulation of immunoreactive beta-endorphin secretion from adult and fetal hypothalamus by sequential stimulation with corticotropin-releasing hormone. Previous work has shown corticotropin-releasing hormone (CRH) stimulation of beta-endorphin (END) secretion from hypothalamus. We tested the hypothesis that CRH stimulation of beta-END (measured by radioimmunoassay) from hypothalamic explants is dependent on: (1) ovine CRH dose, (2) pattern and sequence of CRH stimulation, (3) androgen status, and (4) hypothalamic age. Hypothalami from adult male rats and day 17 fetal rats were studied. In adult hypothalami, CRH-stimulated immunoreactive (IR)-beta-END secretion with 10(-7) M was greater than that with 10(-8) M CRH and showed dose-dependent stimulation. Serial stimulation for 20 min by 10(-8) M CRH followed by a 40 min interval without CRH stimulation resulted in a brief stimulation of secretion of IR-beta-END and also secretion of IR-alpha-melanocyte-stimulating hormone (MSH), another peptide derived from pro-opiomelanocortin, the precursor of beta-END. Subsequent stimulation with 10(-6) M CRH showed a desensitization to stimulation despite readily releasable pools of IR-beta-END shown by potassium-induced depolarization. In addition, prolonged stimulation for 1 h with 10(-7) M CRH or increasing concentrations of CRH produced a sustained increase in IR-beta-END release as long as CRH was present. Dihydrotestosterone treatment had no effect on basal nor CRH-stimulated IR-beta-END release in orchiectomized rats. The pattern of IR-beta-END secretion from fetal hypothalamic explants exposed briefly (20 min) or for a prolonged period (1 h) to CRH was similar to that from adult explants. These results demonstrate that: (1) CRH-stimulated IR-beta-END secretion from hypothalamus is dose-dependent.(ABSTRACT TRUNCATED AT 250 WORDS)


The GABAB receptor antagonist, CGP-35348, inhibits paired-pulse disinhibition in the rat dentate gyrus in vivo. Extracellular field potentials were recorded from the dentate gyrus of adult rats during electrical stimulation of the angular bundle in vivo. Paired pulses produced inhibition of the second population spike (PS2) at 25 ms, and potentiation at inter-stimulus intervals from 50 to 200 ms. The GABAB receptor antagonist, CGP 35348, reduced the amplitude of PS2 at each of these inter-stimulus intervals while producing no effect on the first population spike of the pair (PS1). Under control conditions, the duration of the second population EPSP (pEPSP2) was increased relative to the first at inter-stimulus intervals from 100 to 400 ms, and CGP 35348 reduced these durations to baseline levels. These results demonstrate that GABAB receptors modulate synaptic inhibition in vivo.


Role of hypothalamic paraventricular nucleus alpha- and beta-adrenergic receptors in regulation of blood glucose, free fatty acids and corticosterone. The potential roles of adrenergic and noradrenergic terminals in the hypothalamic paraventricular nucleus in the regulation of blood glucose and free fatty acids, the two major metabolic fuels, were examined. Corticosterone was also measured, both to assess the specificity of any effects for metabolic fuels, and because endogenous catecholamines in this site have previously been implicated in corticosterone regulation. In the first experiment adult male albino rats having chronically implanted guide cannulae aimed at the hypothalamic paraventricular nucleus or the caudate nucleus received microinjections of the agonists methoxamine (alpha 1), clonidine (alpha 2), and isoproterenol (beta) (0, 10, 30, 100 nmol/500 nl), and blood samples were taken from the tail tip. In the second experiment a different set of rats received 30 nmol clonidine or vehicle subcutaneously instead of brain microinjections. Intracranial clonidine and isoproterenol produced marked and moderate hyperglycemia, respectively; methoxamine did not alter glucose. For neither clonidine nor isoproterenol was there any difference in hyperglycemia as a function of microinjection site; also, subcutaneous clonidine injections produced the same peak glucose response as was found after both paraventricular and caudate nucleus microinjections of the same dose. Free fatty acid levels were increased by clonidine and isoproterenol, but slightly suppressed by methoxamine; the alpha agonist effects, but not the beta agonist effect, were greater after paraventricular microinjections than after caudate microinjections. Corticosterone was increased by both alpha agonists after paraventricular but not after caudate nucleus microinjections; beta agonist microinjections into the paraventricular and caudate nuclei produced equivalent corticosterone elevations. These results suggest that most, if not all, of the hyperglycemic effects of alpha and beta adrenergic agonist microinjection into the paraventricular nucleus can be ascribed to leakage of the material into the vasculature, with subsequent action at a distant site. In contrast, all 3 agonists seem capable of acting within the brain to alter free fatty acid levels. The effects on corticosterone of both the alpha 1 and alpha 2 agonists, but not the beta agonist, also appear due, at least in part, to actions within the brain. Previous suggestions that catecholamine terminals in the hypothalamic paraventricular nucleus are directly and strongly involved in metabolic fuel regulation may require reconsideration.


Electrophysiological properties of early postnatal rat septal neurons in short-term culture. Electrophysiological properties of septal neurons dissociated from PN1-PN7 rats were examined between 1 and 5 days in vitro (DIV) with whole-cell patch-clamp recording. The neurons had RMPs in the range -40 to -80 mV, resistances 0.5-1.5 G omega, and 50-90 mV action potentials. By 2-3 DIV, most neurons were spontaneously active, with some cells exhibiting rhythmic firing patterns. Depolarizations from -80 mV holding potential elicited TTX-sensitive inward Na+ currents, and transient and sustained outward K+ currents. Pharmacological dissection of the outward currents in PN6/7 neurons suggest the presence of multiple types of K+ currents (IA, IC, IK). L-type Ca2+ currents were observed in all PN6/7 neurons examined but were not always detectable in PN1/2 neurons. All PN1/2 and PN6/7 neurons were sensitive to glutamate and GABA but did not respond to ACh or NE applications. Responses to GABA were excitatory i.e., characteristic of immature neurons. Comparison of the results obtained in this study with the properties of adult neurons characterized in vivo or in brain slice preparations in vitro, suggest that septal neurons from PN1-PN7 rats are still in the process of differentiation of their mature electrical and chemosensitive membrane properties.


Osmotic stimulation induces changes in the expression of beta-adrenergic receptors and nuclear volume of astrocytes in supraoptic nucleus of the rat. The influence of osmotic stimulation on the density of beta-adrenoceptor binding sites in the rat supraoptic nucleus (SON) was studied by quantitative autoradiography using 125I-cyanopindolol (ICYP). Increased density of beta-adrenoceptor binding sites was observed in osmotically stimulated rats and also after the suppression of neuronal activation by rehydration of animals. This was mainly due to a significant increase in the concentration of beta 2 binding sites. The overexpression of beta-adrenoceptors occurred concomitantly with nuclear expansion in SON astrocytes. Moreover, the higher concentration of beta-adrenoceptors observed in the ventral portion of the SON largely coincided with the area that showed intense GFAP-immunostaining. These results provide indirect evidence of an astrocytic location of beta-adrenoceptors and also of beta-adrenergic mediation in the structural and functional changes of SON astrocytes.


Functional neuronal binding sites for oxytocin in the ventromedial hypothalamus of the guinea pig after gonadectomy. Castration in rats of either sex has been shown to markedly decrease hypothalamic oxytocin binding in the ventromedial hypothalamus and this can be reversed by injecting gonadal steroids. We wondered whether castration exerts similar effects on homologous oxytocin binding sites present in the guinea pig hypothalamus. Adult male guinea pigs were castrated and killed 2-90 days later. Binding sites for oxytocin in the ventromedial nucleus and neuronal responses to this peptide were little affected by gonadectomy, in contrast to what is observed in the rat under similar experimental conditions. The steroid dependency of hypothalamic oxytocin receptors appears therefore to be species dependent.


Interaction of natriuretic peptides and cGMP production via the same receptor in mouse astrocytes. Atrial and brain natriuretic peptides have been found previously to bind to specific receptors on cultured mouse astrocytes and to stimulate cyclic guanosine 5-monophosphate (cGMP) production with similar dose dependency although brain natriuretic peptide (BNP) shows a greater maximal stimulatory effect. The present study provides evidence that both peptides work through the same pathway. No additive or synergistic effect was observed when astrocytes were exposed to both peptides. However, human ANF(99-126) at high concentrations partially inhibited porcine BNP induced cGMP production to the level seen with ANF alone. ANF could be viewed as a partial agonist of pBNP competing for the same effector sites. Differences in structure between human ANF(99-126) and porcine BNP may account for the difference in cGMP response. The interaction between the two peptides and the cGMP response does not reflect receptor binding affinities and is likely to be a post-binding event.


Central opioid receptor subtype antagonists differentially alter sucrose and deprivation-induced water intake in rats. The present study compared the effectiveness of centrally-administered opioid receptor subtype antagonists to inhibit intake of either a 10% sucrose solution under ad libitum conditions, or water following 24 h of water deprivation. Full dose-response functions were evaluated over a 1 h period for the following antagonists: naltrexone (general: 1-50 micrograms), nor-binaltorphamine (Nor-BNI, kappa: 1-20 micrograms), beta-funaltrexamine (beta-FNA, mu: 1-20 micrograms), naltrindole (delta 2: 1-20 micrograms), [D-Ala2, Leu5, Cys6]-enkephalin (DALCE, delta 1: 10-40 micrograms) and naloxonazine (mu 1: 10-50 micrograms). Naltrexone significantly and dose-dependently inhibited both sucrose intake (64-67%) and deprivation-induced water intake (53-67%). Nor-BNI significantly and dose-dependently inhibited sucrose intake (53-55%), but failed to significantly affect (28%) deprivation-induced water intake. beta-FNA significantly and dose-dependently inhibited both sucrose intake (31-34%) and deprivation-induced water intake (36-50%). Naltrindole failed to significantly alter either sucrose intake (24%) or deprivation-induced water intake (16%). Whereas DALCE significantly, but transiently (15-20 min) inhibited sucrose intake (28%), it failed to significantly alter deprivation-induced water intake (14%). Naloxonazine significantly, but transiently (5-10 min) stimulated sucrose intake at low doses (26%), but non-significantly reduced sucrose intake at higher doses (20%). Naloxonazine failed to significantly alter deprivation-induced water intake (16% reduction). These data indicate that whereas the kappa and mu 2 binding sites participate in the opioid modulation of sucrose intake, the mu 2 binding site participates in the opioid modulation of deprivation-induced water intake.


Activity of the hypothalamic-pituitary-adrenal axis in mice selected for left- or right-handedness. Asymmetry in brain modulation of the immune system has been previously described. In mice, paw preference has been shown to be associated with immune reactivity but the mechanisms involved in such an association are not yet known. The autonomic nervous system and the neuroendocrine system are considered as major candidates for neural influences on the immune system. In the present study, the activity of the hypothalamic-pituitary-adrenal (HPA) axis of adult female mice selected for paw preference (left-handers vs. right-handers) was assessed by measuring both adrenocorticotropic hormone (ACTH) and corticosterone plasma levels, as well as the in vitro responses of hypothalamus and adrenocortical cells to various hormone releasing stimuli. The results reported here showed no difference in the activity of the HPA axis between left- and right-handed mice, suggesting that this neuroendocrine axis is not implicated in the association between functional brain asymmetry and immune reactivity. However, our results do not exclude the possibility that the HPA axis could play a role in such an association under other circumstances, such as during development or stressful situations.


A correlative study of calcium channel antagonist binding and local neuropathological features in the hippocampus in Alzheimer's disease. [3H]PN200-110 binding sites were studied by means of quantitative autoradiography in hippocampal sections of patients with Alzheimer's disease and age-matched control subjects. Choline acetyltransferase activity, plaque, tangle and cell densities were also determined in the same tissue samples used for autoradiographic studies. Quantitative autoradiographic analysis of [3H]PN200-110 binding in control hippocampus revealed a heterogeneous pattern similar to that described in rodents, being particularly high in the dentate gyrus. In Alzheimer's disease, [3H]PN200-110 binding was markedly reduced in the subiculum (control = 9.85 +/- 1.41 pmol/g; Alzheimer = 3.41 +/- 0.54 pmol/g, mean +/- S.E.M., P less than 0.001). In the subiculum there was a disproportionate reduction of [3H]PN200-110 binding in comparison to cell loss in Alzheimer's disease. The activity of choline acetyltransferase in the hippocampus was markedly reduced in Alzheimer's disease (controls 6.9 +/- 1.0; Alzheimer 2.7 +/- 0.9 nmol/h/mg protein, mean +/- S.E.M., P less than 0.01). There was a strong correlation between choline acetyltransferase activity and [3H]PN200-110 binding in the subiculum. [3H]PN200-110 binding did not correlate with plaque density in the subiculum. The discrete reduction and preservation of [3H]PN200-110 binding in the present study is consistent with the pattern of selective cellular vulnerability in the hippocampal region in Alzheimer's disease.


Mouse telencephalon exhibits an age-related decrease in glutamate (AMPA) receptors but no change in nerve terminal markers. The central excitatory amino acid receptor selective for alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) was examined in brain tissue from mice at 3 and 25 months after birth. Antibodies against the rat GluR-A glutamate receptor subunit (selective for kainate and AMPA) labeled a mouse brain component of about M(r) 100,000. Telencephalic tissue from the older group of mice exhibited 31% less immunoreactivity towards this component as compared with that from the young group. Binding of [3H]AMPA also decreased with age in the telencephalon to an extent which was similar to the loss of receptor immunoreactivity. Scatchard analysis revealed that this reduction is due to a decrease in receptor density and not to a change in binding affinity. In contrast, there were only small age-related changes in AMPA receptor immunoreactivity and binding levels in the brain stem and cerebellum. Binding to dopamine, serotonin, or GABA receptors was not significantly reduced in the older mice. Since the nerve terminal markers synaptophysin and the SV2 glycoprotein were not detectably different in the two groups of mice, the age-related reduction in AMPA receptors is not likely to be due to a general decrease in synaptic density. These data suggest that glutamatergic neurotransmission mediated by AMPA-type receptors is selectively impaired with aging in the telencephalon.