Fimbriation, hemagglutination and adherence properties of fresh clinical isolates of Branhamella catarrhalis.

This study investigated the fimbriation on 24 fresh clinical isolates of Branhamella catarrhalis by electron microscopy. All the strains were isolated from patients with respiratory infections. The Branhamella catarrhalis strains were classified into three groups according to the grade of fimbriation. Among these 24 strains the incidence of densely fimbriated, moderately fimbriated and sparsely fimbriated isolates were 12 (50%), 7 (29%) and 5 (21%), respectively. After five-times serial subculture on Brain Heart Infusion agar, the average number of fimbriae per bacteria was decreased from 174 to 114 in the densely fimbriated strain and from 48 to 10 in the moderately fimbriated strain. Moreover, 20% of the population became non-fimbriated in moderately fimbriated strain after the serial subculture. In strains with higher hemagglutination titer the number of fimbriae was significantly (P < 0.04) more than in strains with lower hemagglutination titer.


Role of protein kinase C in the modulation of multidrug resistance: expression of the atypical gamma isoform of protein kinase C does not confer increased resistance to doxorubicin.

Cross-resistance to anticancer drugs, termed multidrug resistance (MDR), is functionally associated with the expression of a plasma membrane, energy-dependent, drug efflux pump termed P-glycoprotein (PGP), the product of the mdr1 gene. We have shown previously that MCF-7 breast carcinoma cells transfected with the human mdr1 gene (BC-19 cells) exhibit greater MDR when stably transfected with protein kinase C alpha (PKC alpha). We now demonstrate that transfection of BC-19 cells with the gamma isoform of PKC (BC-19/PKC gamma cells), which is not normally present in BC-19 cells, does not confer increased resistance to doxorubicin, despite a 19-fold increase in PKC activity. All of the increased PKC activity is accounted for by PKC gamma and it is rapidly down-regulated by phorbol dibutyrate, within 15 min of treatment. Endogenous PKC alpha and PKC epsilon activities are not affected by phorbol dibutyrate. The cytotoxicity of doxorubicin was similar in BC-19/neo or BC-19/PKC gamma cells after either 2-hr or continuous drug exposure, and co-treatment with phorbol dibutyrate increased resistance to doxorubicin 4-fold in both cell lines. Phosphorylation of PGP was similar in both cell lines and drug accumulation was not affected by overexpression of PKC gamma. These results demonstrate that transfection of PGP-expressing cells with an atypical isoform of PKC does not confer increased MDR, and they suggest that the regulation of PGP is phenotype specific with respect to the isoform of PKC.


Molecular cloning and expression of a pituitary somatostatin receptor with preferential affinity for somatostatin-28.

Using the polymerase chain reaction technique with degenerative primers, we obtained from a rat pituitary cDNA library a cDNA fragment, rAP236, that exhibited considerable homology to known receptors that belong to the guanine nucleotide-binding protein (G protein)-coupled receptor superfamily. Oligonucleotides to this fragment were used as probes to obtain a full-length cDNA from the rat pituitary cDNA library. This clone, rAP6-26, encoded a 383-amino acid protein with seven putative transmembrane domains that are characteristic of G protein-coupled receptors. The predicted amino acid sequence of the rAP6-26 cDNA exhibits 56-66% homology to recently cloned somatostatin (SRIF) receptors. Membranes prepared from COS-7 cells transfected with the rAP6-26 cDNA showed specific binding of 125I-Tyr11-SRIF, thus identifying the cDNA clone as a novel SRIF receptor. Radioligand binding competition analysis using somatostatin-28 (SRIF-28) and a number of cyclic SRIF analogs revealed that SRIF-28 was the most potent competitor of 125I-Tyr11-SRIF binding, with a approximately 30-fold greater affinity for the receptor than that of SRIF. In addition, binding of 125I-Tyr11-SRIF was markedly reduced in the presence of Na+ ions and GTP, indicating coupling of rAP6-26 receptors to inhibitory G proteins in COS-7 membranes. In adenylyl cyclase assays, forskolin-induced cAMP accumulation was inhibited by SRIF and SRIF-28, thus confirming that the rAP6-26 cDNA encodes a functional receptor protein. By Northern blot analysis, a approximately 2.6 kilobase mRNA encoding the receptor was present in the pituitary but not in the liver, small intestine, kidney, pancreas, cerebellum, or cortex. Lack of receptor mRNA expression in the brain was confirmed by in situ hybridization histochemical studies. Thus, we report the cloning of a novel rat pituitary SRIF receptor, termed SSTR4, that has marked preferential affinity for SRIF-28 and is linked to inhibition of adenylyl cyclase.


Inhibition by HS-142-1, a novel nonpeptide atrial natriuretic peptide antagonist of microbial origin, of atrial natriuretic peptide-induced relaxation of isolated rabbit aorta through the blockade of guanylyl cyclase-linked receptors.

HS-142-1, a specific nonpeptide antagonist for the atrial natriuretic peptide (ANP) receptor, equally blocked rat ANP (rANP)-, porcine brain natriuretic peptide-, or porcine C-type natriuretic peptide-stimulated GMP production in cultured bovine aortic smooth muscle (BASM) and bovine aortic endothelial (BAE) cells in a concentration-dependent fashion, at concentrations of 1-300 micrograms/ml. But, even at 300 micrograms/ml, HS-142-1 only weakly inhibited the specific binding of 125I-rANP to the BASM and BAE cells, where only a small portion of the binding sites are linked to guanylyl cyclase. Further, with BAE cell membranes, HS-142-1 recognized only the 135-kDa ANP receptor, which is thought from 125I-rANP affinity cross-linking studies to be the guanylyl cyclase-linked receptor. HS-142-1 also, if anything, inhibited the labeling of 135-kDa ANP receptors in the affinity cross-linking studies with BASM membranes, suggesting that a major portion of the 135-kDa ANP receptors are HS-142-1 insensitive and only a small portion of the 135-kDa ANP receptors are responsible for the blockade by HS-142-1 of GMP production in BASM cells. At a concentration of 100 micrograms/ml, HS-142-1 reversibly prevented ANP-induced relaxation of the isolated rabbit thoracic aorta induced to contract with 3 x 10(-7) M phenylephrine, but not the relaxation induced by sodium nitroprusside, isoproterenol, or papaverine. These results suggest that HS-142-1 specifically inhibits natriuretic peptide-induced vasorelaxation through the blockade of guanylyl cyclase-linked natriuretic peptide receptors. HS-142-1 thus will be a powerful tool for understanding the physiological roles, in vasculature, of natriuretic peptides, which contribute to the homeostasis of blood pressure and intravascular volume.


Studies on the interaction of leucocytes and the myocardial vasculature. I. Effect of hypoxia on the adherence of blood granulocytes.

Granulocytes play an important role in increasing the infarct size after ischemia and reperfusion by the release of oxygen-derived free radicals (ODFR) and lysosomal enzymes. It has been shown that the number of granulocytes adhering to the vascular endothelium increases after occlusion of the coronary artery, and that the area of myocardial damage can be reduced by preventing granulocyte adherence with monoclonal antibodies directed against adhesion receptors. The underlying mechanism of granulocyte activation under these conditions is not yet known. We have investigated whether granulocytes can be activated directly by reduced oxygen tensions. Granulocytes were suspended in a hypoxic buffer and incubated on fibronectin and gelatin coated microtitre plates at 1-3% ambient oxygen to study their ability to adhere to these matrices. The results showed that the adherence of granulocytes to fibronectin was dependent on the duration of hypoxia. After 30 min of incubation under hypoxia granulocyte adherence increased 1.3 to 1.8 fold compared to the normoxic control. The adherence to fibronectin could be inhibited partially by anti-CD18 antibody, a monoclonal antibody to the common beta chain of a class of extracellular matrix receptors. This direct activation of granulocytes due to hypoxic conditions may have implications for the interaction of these cells with the vascular endothelium in vivo.


Single-strand conformation polymorphism (SSCP) analysis applied to the diagnosis of acute intermittent porphyria.

The single-strand conformation polymorphism (SSCP) technique was used to detect carriers of the known point mutation in the first exon of the porphobilinogen deaminase gene in Finnish and Swedish families. The SSCP technique was a reliable and convenient way of distinguishing patients from healthy members in a family. This point mutation is thought to result from a splicing defect of the mRNA. The PCR-based analyses of a patient's cDNA did not reveal the presence of an abnormal mRNA population, suggesting that no abnormal mRNA is synthesized or that it is too unstable to be detected.


Occurrence and functions of the phosphatidylinositol cycle in the myocardium.

In the last decade a great deal of attention was awarded to a signal transduction pathway which is utilized primarily by 'Ca2+ mobilizing' signal molecules and which involves the hydrolysis of a quantitatively minor phospholipid, phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) by a PtdIns-specific phospholipase C (PLC). The evidence for the existence of receptor-mediated GTP binding protein-coupled PLC in myocardium and its possible functions are briefly summarized. The minireview is concentrated on the following aspects: 1) cellular localization and synthesis of polyphospho-PtdIns from PtdIns, 2) desensitization of the alpha 1-adrenergic agonist and endothelin-1 mediated PtdIns responses, 3) oscillatory Ca2+ transients initiated by PtdIns(4,5)P2 hydrolysis, 4) polyunsaturated fatty acids as constituents of polyphospho-PtdIns and of the protein kinase C activator 1,2-diacylglycerol (DAG), 5) source other than PtdIns(4,5)P2 contributing to the stimulated DAG, 6) role of the PtdIns pathway in cardiomyocyte growth and gene expression during the hypertrophic response.


Application of magnetic resonance angiography to neurosurgical practice: a critical review of 150 cases.

The potential role of magnetic resonance angiography (MRA) in clinical neurosurgical practice was evaluated in 150 consecutive patients in an attempt to define clinical situations in which it aided in diagnosis, helped guide treatment options, and/or possibly eliminated the need for conventional catheter angiography. Among patients with nonvascular pathology (n = 42), MRA provided useful clinical information in 55% of cases, and could have effectively replaced catheter angiography in 76% of cases. In this group of patients, MRA provided excellent depiction of tumour and vessel relationships, and in conjunction with conventional imaging modalities reliably ruled out the question of tumour versus aneurysm whenever that question was raised. Among patients with vascular lesions (n = 108), MRA visualized the lesion in 18 of 20 (90%) aneurysms, 11 of 11 (100%) vascular malformations, 31 or 31 (100%) cases with known occlusive vascular disease, and reliably excluded occlusive vascular disease in 19 of 30 (63%) cases. The MRA potentially could have replaced conventional catheter angiography in less than 8% of cases with vascular pathology treated surgically, and in 76% of cases with vascular pathology treated medically or expectantly. Imaging time was relatively short, 10 to 20 minutes for intracranial vessels and 6.5 to 13 minutes for extracranial vessels. Shortcomings included the lack of spacial resolution required for surgical planning and the possible misleading information in the settings of low flow and partial thrombosis. It is concluded that currently available MRA modalities contribute useful information in a wide spectrum of neurosurgical clinical situations including screening and serial follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)


Differential expression of heat shock protein 70 gene between the cortex and caudate after transient focal cerebral ischaemia in rats.

In relation to changes of total protein synthesis, induction of 70-kDa heat shock protein (HSP70) mRNA was examined by Northern blot and in situ hybridization after 30 min of transient middle cerebral artery (MCA) occlusion of rats. HSP70 mRNA was not present in the control condition of brain. With reperfusion, the mRNA was greatly induced along with the recovery of total protein synthesis in the cerebral cortex and lateral caudate of the ipsilateral hemisphere. However, the level of the mRNA reached a maximum earlier in the lateral caudate (at 3 h) than in the cortex (at 8 h), and the maximum amount of the mRNA was much smaller in the caudate than in the cortex. Total protein synthesis in the lateral caudate did not completely recover until 7 days. Histological examination showed a severe damage in cells of lateral caudate, while cells in the cortex were almost normal at 7 days. No difference in the brain temperature was observed between the two regions. These results show that the induction of HSP70 mRNA correlates with the recovery of protein synthesis in brain cells after a transient ischaemia, and that the HSP70 gene expression is different at the transcriptional level between the cortical and caudate cells after the transient ischaemia.


Introduction of high definition television system to neurosurgical documentation.

The high definition television (HDTV) system is introduced to microneurosurgery. Five cases with intracranial lesions: three cerebral aneurysms (giant aneurysms of the anterior communicating artery and internal carotid artery, aneurysm of the basilar artery), one acoustic neurinoma and one skull base meningioma, were operated on under the microscope using the HDTV system. The surgical procedures of each case were relayed and recorded by the system. In two of the five cases, we used two sets of HDTV system to produce stereoscopic projection. The HDTV system provided us with images of superior quality with a distinctly greater resolution than ordinary video systems.


Morphological quantitative analysis of intracranial pressure waves in normal pressure hydrocephalus.

This work presents a prospective morphological and quantitative analysis of 43 intracranial pressure recordings performed on normal pressure hydrocephalic patients. This analysis led us to separate Lundberg's B waves into different subtypes and to refine the definition of the 'Plateau' wave. Two B wave subtypes named Great Symmetrical wave and Intermediate wave appeared correlated with the surgical improvement. In addition, the degree of post-operative improvement was correlated with the frequency of Intermediate wave. An extended quantitative classification of intracranial pressure waves is proposed that can be used alone to determine which patients should undergo a shunting procedure and which one should the most improve.


Activation of phospholipase D by platelet-derived growth factor (PDGF) in rat C6 glioma cells: possible role in mitogenic signal transduction.

The effects of platelet-derived growth factor (PDGF) on phospholipase D (PLD) activity and deoxyribonucleic acid (DNA) synthesis in rat C6 glioma cells have been investigated. Pretreatment of serum-starved C6 cells with PDGF results in enhanced choline production and the phosphatidylethanol (PEt) formation in the presence of ethanol, indicating the activation of PLD acting on phosphatidylcholine (PC). The dose-response curve for choline generation and DNA synthesis were comparable. In addition, the effects of PDGF on both PEt formation and [3H]thymidine incorporation into acid-precipitable material was blocked by the potent protein kinase C (PKC) inhibitor 1-(5-isoquinolinesulphonyl)-2-methylpiperazine (H-7) but not by N-(2-guanidinoethyl)-5-isoquinolinesulphonamide (HA1004), a relatively weak inhibitor of PKC, suggesting that PDGF plays an important role as a positive regulator of glioma cell growth via a PLD-mediated mitogenic signal transduction cascades, which depends largely on the activation of PKC.


Leukotriene C4 contents, synthase and catabolic activity in human meningiomas.

Leukotriene has been proposed as a factor of tumour induced brain oedema. Independently of its size, meningioma occasionally shows various extents of peritumoural oedema. We investigated LTC4 tissue contents, LTC4 catabolic and synthetic activity in 12 human meningiomas and their correlation with peritumoural oedema was studied. LTC4 contents were varied from 0.01 to 8.21 pg/mg tissue. When LTA4, an unstable expoxide intermediate was incubated with tumour homogenate, LTC4 was rapidly synthesized. However, LTC4 levels generated by incubating LTA4 with each homogenate were much different in each case. Degradation of LTC4 to LTD4, LTE4, and other polar materials was also rapid by incubation with tumour homogenates. Approximately 70% of added LTC4 was transformed to LTD4, LTE4 nor 6-trans LTB4 diastereoisomers during 30 min incubation at 37 degrees C. The results suggested that there were significant LTC4 tissue contents and LTC4 synthetic and catabolic activity in meningiomas. Oedema index ranged from 1.0 (no peritumoural oedema) to 67.5. No significant correlation, however, was observed not only between the LTC4 tissue contents and LTC4 synthetic or catabolic activities but also between each of these three parameters and peritumoural oedema. Thus, these results do not support a significant correlation of sulfidopeptide LTs with oedema formation in meningioma patients. Since leukotrienes are extremely unstable compounds, LTC4 tissue contents should be carefully discussed along with a consideration of rapid LTC4 synthesis and catabolism. Further role of leukotrienes in meningioma tissue should be studied.


Reversibility of energy metabolism and intracellular pH after cerebral ischaemia evaluated by 31P-MRS.

We have previously developed a reproducible model of transient forebrain ischaemia in rats by bilateral carotid artery occlusion combined with temporary increase of ICP. With this model, reversibility of the energy metabolism and intracellular pH (pHi) was investigated by 31P-MRS during 120 min of recirculation in three groups of, respectively, 30, 60, and 120 min of ischaemia. With the induction of ischaemia, ATP and phosphocreatine (PCr) disappeared, and measurement of pHi showed severe acidosis in all rats. In the 30 min ischaemia group, both energy metabolism and pHi recovered almost completely. In the 60 min ischaemia group, ATP recovered to 74% of control values, but pHi showed full recovery. In the 120 min ischaemia group, ATP recovered to about 50% of control values, and recovery of pHi was variable. Showing logarithmical changes during recirculation in ATP and PCr, the rate of metabolic recovery was fast during 60 min of recirculation, but it decreased and reached plateau thereafter in all groups. Recovery of pHi was affected by ATP levels, and was precipitously accelerated as ATP levels exceeded 50% of pre-ischaemic values. These results suggest that prolongation of the duration of ischaemia limits the restoration of the energy state, and the quality of pHi recovery after cerebral ischaemia is affected by the degree of ATP recovery during 60 min of recirculation.


Cerebral venous system mechanics during a constant infusion in subarachanoid space: a model study.

A model has been developed to explain the development of stress under a constant infusion of fluid into the subarachanoid space. When the regulatory mechanism operates, the infused fluid is accommodated by the venous system and hence the tissue 'give' mechanism is under minimal stress. The necessity of maintaining a constant blood flow causes the blood vessel to dilate at higher CSF pressures. This puts the tissue 'give' mechanism under dual pressures, since it has to make room for the expanding ventricle as well as the dilating venous system. The theoretical model developed has been validated by experimental observations.


Fenestration of the middle cerebral artery and aneurysm at the site of the fenestration.

The authors report the case of a patient who presented a ruptured aneurysm of the anterior communicating artery and an unruptured aneurysm of the middle cerebral artery arising at the site of a fenestration of the MCA. The fenestration was undiagnosed on the preoperative angiogram but discovered during the surgery carried out for clipping of the aneurysms. In the literature, cases of fenestration of the MCA are sporadically reported and are incidental findings; an aneurysm may be associated on an artery other than the fenestrated MCA; an aneurysm arising at the site of the MCA fenestration is a very rare occurrence.


Somatostatin is altered in developing retina from ethanol-exposed rats.

Optic nerve hypoplasia is seen in 50% of patients diagnosed with fetal alcohol syndrome and is due to a defect in the development of retinal ganglion cells. Somatostatin may influence the development of retinal ganglion cells, so we studied the effect of in vivo ethanol exposure on somatostatin expression in the developing retina. Somatostatin concentration is increased in retina from ethanol-exposed fetuses and pups and this increase in peptide is associated with excessive neurite formation and improper migration of the somatostatin-containing neurons at early postnatal ages. These disturbances may account for the eventual failure in retinal ganglion cell development that results in optic nerve hypoplasia.


Responses to repetitive afferent activity of rat solitary complex neurons isolated in brainstem slices.

The response of postsynaptic solitary complex neurons to repetitive stimulation (20-50 Hz) of the tractus solitarius were investigated by intracellular recordings in a brainstem slice preparation. Short duration stimuli (0.5 s) elicited increases in synaptic activity and short-term potentiation of synaptic potentials, both of which lasted approximately 1 min, plus a 10 s repolarization suppressed in the presence of glutamate ionotropic receptors antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) and 2-D-amino-7-phosphonoheptanoic acid (AP7, 50 microM). Longer (5 s) stimuli elicited 2-10 min depolarizations accompanied by membrane resistance increases and unaffected by glutamate ionotropic receptors antagonists. Our study reveals several mechanisms by which rhythmic visceral afferents may exert a tonic control of postsynaptic solitary complex neurons.


Somatostatin effects on cultured human fetal epiphyseal chondrocytes.

Somatostatin effects on cultured human fetal epiphyseal chondrocytes were evaluated by studying the effects of somatostatin on DNA synthesis. Cultured epiphyseal chondrocytes from human fetuses (12-40 wk old) were incubated for 48 h in Ham's F-12 serum-free medium. After this, the medium was replaced by MCDB-104 serum-free medium and the cells were incubated for an additional 48 h in the presence or absence of somatostatin 1 pM to 10 microM, with the addition of 3H-thymidine (5 microCi/mL) for the last 24 h of incubation. A significant (p < 0.02) inhibitory effect of somatostatin (1 nM to 10 microM) on 3H-thymidine DNA incorporation was observed in cultured chondrocytes from fetuses of all gestational ages studied (12-40 wk), with no significant differences among fetal ages. In conclusion, our results show that somatostatin exerts a biologic effect on cultured human fetal epiphyseal chondrocytes, as it does in its target cells. These results suggest that somatostatin could regulate human skeletal growth not only by growth hormone secretion regulation, but also by acting directly on chondrocyte metabolism. However, the physiologic significance of the latter remains to be elucidated.


Astra Award Lecture. The psychopharmacology of 5-HT3 receptors.

5-HT3 receptors have an exclusive neuronal location and evidence is presented of their involvement in behaviour. 5-HT3 receptor antagonists such as ondansetron, tropisetron and zacopride have provided the critical pharmacological tools to reveal a potent and efficacious ability to regulate disturbed behaviour. Thus the 5-HT3 receptor antagonists will restore to normal rodent and primate behaviour disturbed by increasing limbic dopamine function, aversive situations, cognitive impairments and drug abuse. The remarkable feature of their action is a failure to modify normal behaviour. This unique pharmacological signature has ensured a wide interest in the potential role of the 5-HT3 receptor antagonists in the treatment of schizophrenia, anxiety, age related memory impairment and the problems of withdrawal from drugs of abuse. The preclinical data and preliminary clinical observations are presented.


Calcium entry blockade as a mechanism for chlordimeform-induced inhibition of motor activity in the isolated guinea-pig ileum.

Central and peripheral alpha 2-adrenoceptors, including those of the gastrointestinal tract, have been indicated as a toxicity target of formamidine pesticides in mammals. In this study, the inhibitory effect of chlordimeform on twitch contractions from electrically-stimulated longitudinal muscle-myenteric plexus preparations (LMMPs) of the guinea-pig ileum was found to be resistant to the action of the alpha 2-adrenoceptor antagonist idazoxan. This drug was also ineffective on chlordimeform-induced inhibition of peristalsis recorded in whole ileal segments. As expected, idazoxan antagonized the inhibitory effect of the alpha 2-adrenoceptor agonist clonidine on twitch contractions and peristaltic activity. Chlordimeform reduced the amplitude of direct mechanical responses to a variety of spasmogens such as acetylcholine, histamine and substance P, suggesting a muscular site of action. Moreover, Ca(2+)-free, K(+)-depolarized LMMPs, chlordimeform inhibited submaximal contractions caused by addition of exogenous calcium, through an action apparently similar to that of the Ca2+ entry blocker nifedipine. Both chlordimeform- and nifedipine-induced inhibition of calcium contractions were reversed by the calcium channel activator BAY K 8644. This compound also partially prevented the inhibitory action of chlordimeform on peristaltic activity. On the whole, these results indicate that chlordimeform-induced depression of motor activity in the guinea-pig ileum is, at least in part, related to inhibition of transmembrane Ca2+ fluxes responsible for smooth muscle contraction.


The antibacterial effect of some neuroleptics on strains isolated from patients with meningitis.

Eighty-two strains of bacteria (Neisseria meningitidis, Haemophilus influenzae, Enterobacteriaceae, Streptococcus pneumoniae, group B streptococci and Listeria monocytogenes) were examined for their in vitro susceptibility to eight drugs, seven neuroleptics (perphenazine, fluphenazine, cis(Z)-clopenthixol, haloperidol, clozapine, clebopride and SCH 23390), and the neuroleptically inactive trans(E)-clopenthixol. The phenothiazines and the thioxanthenes were, on the whole, the most active drugs when measured, the IC50(50) for each group of bacteria being 7.4 to 84 mg/l (with the exception of the activity against the enterobacteriaceae). The antibacterial potency of clozapine, which has an atypical neuroleptic profile, was between 50 and 140 mg/l. Haloperidol also showed an antibacterial activity in the concentration range 35-140 mg/l. The selective D1 antagonist, SCH 23390 and the selective D2 antagonist, clebopride, inhibited only few of the bacteria in the concentration range investigated.


Selectivity and potency of opioid peptides in regulating human chorionic gonadotropin release from term trophoblast tissue.

The in vitro effect of three opioid peptides on hCG release from term trophoblast tissue was investigated. These peptides were prototypic for opioid receptors of the following types: kappa [dynorphin(1-13)], mu (DAMGO) H-Tyr-D-Ala2-Gly-N-Me-Phe-Gly5- ol, and delta (DPDPE) H-Tyr-O-Pen-Gly-Phe-D-Pen-OH[D-Pen2,D-Pen5]enkephalin. All peptides stimulated hCG release and their concentration-response curves were bell shaped. Their order of potency was kappa >>> mu > delta. Stimulation of hCG release by any of the peptides was totally reversed by opioid antagonists, indicating that the action of peptides is mediated by placental opioid receptors. In order to confirm the specificity of opioid regulation of hCG release, three nonopioid drugs (cocaine, nicotine, and isoproterenol), with binding proteins and receptors known to be present in trophoblast tissue membranes, were also investigated. Stimulation of hCG release caused by certain concentrations of nonopioid drugs was not reversed by opioid antagonists, demonstrating that their effect is not mediated by opioid receptors. Furthermore, the concentration-response curve of isoproterenol was biphasic, suggesting the presence of a mechanism regulating hCG release that is not mediated by placental beta-adrenergic receptors. Data presented in this manuscript indicate that placental opioid receptors mediate one of the mechanisms regulating hCG release from trophoblast tissue and confirm our earlier results using opioid drugs.


beta-adrenoceptors in brain and pineal from depressed suicide victims.

beta-Adrenoceptors were measured by saturation binding of [3H]CGP 12177 in nine brain regions and pineal from suicides, with a firm retrospective diagnosis of depression, and age and sex matched controls. Twenty one suicides had not recently received antidepressant drugs, 17 had been receiving drugs prior to death. In antidepressant drug-free suicides, the number of total beta-adrenoceptors was significant lower in temporal cortex (Brodmann area 38) and beta 1-adrenoceptors (Brodmann areas 21/22) was significant lower than matched controls. Suicides who died by violent means had significantly lower numbers of total beta- and beta 1-adrenoceptors in the frontal cortex and lower numbers of beta 1-adrenoceptors in temporal cortex (Brodmann areas 21/22) than matched controls. Suicides who died by non-violent means had lower numbers of total beta-adrenoceptors in occipital cortex controls and lower numbers of total beta- and beta 1-adrenoceptors in temporal cortex (Brodmann area 38) than matched controls. In antidepressant drug-treated suicides, significantly lower number of beta-adrenoceptor binding sites were found in temporal cortex (Brodmann area 38) and thalamus compared to matched controls. The lower number of beta-adrenoceptors binding sites in the thalamus appeared to be related to drug treatment. There were no differences in beta-adrenoceptor binding in the pineal gland between antidepressant-free and antidepressant-treated suicides and controls, although there were apparent differences between suicides and controls related to the time of death and season of death.


Biologic, pharmacologic, and psychosocial factors influencing response to neuroleptics.

Neuroleptic drugs remain a critical modality in the treatment of schizophrenia. Numerous variables influence response to neuroleptic drugs including biologic, pharmacologic, and psychosocial factors. This review cites some examples of how such factors have been examined.


Tardive dyskinesia and diabetes mellitus.

Two studies examine the prevalence of tardive dyskinesia (TD) in neuroleptic-treated diabetic patients. Study 1 compared 38 diabetic patients with 38 nondiabetic patients treated for psychotic disorders with low to moderate doses of neuroleptics (mean chlorpromazine equivalents = 300 mg/day) for an average of 18 years. Study 2 compared 24 diabetic and 27 nondiabetic patients treated for an average of 2.6 years with a mean 31 mg/day of metoclopramide for gastrointestinal disease. Patients were examined for TD using standardized scales by raters blind to all treatment and illness variables. In both studies, there were no differences between the diabetic and nondiabetic groups in age, sex, type of psychiatric illness, and dose and duration of neuroleptic treatment or severity of parkinsonism. In both studies, the diabetic patients had significantly greater prevalence and severity of TD. No measures of diabetes severity were associated with TD in either study. Possible pathophysiologic mechanisms for the increased prevalence of TD in neuroleptic-treated patients with diabetes will be discussed.


Benzodiazepine augmentation of neuroleptics in treatment-resistant schizophrenia.

A significant minority of patients with schizophrenia fail to respond to neuroleptic medication alone. In some of these patients, adjunctive treatment with benzodiazepines may prove beneficial. Preclinical studies suggest that benzodiazepines significantly decrease brain dopamine release and turnover, perhaps by augmenting gamma-aminobutyric acid (GABA) inhibition of dopamine neuron activity. Double-blind clinical studies, however, have not conclusively established a role for benzodiazepines in the treatment of schizophrenia, and it seems likely that some patients respond favorably, whereas others do not. We review preliminary new observations that approximately half of a group of treatment-resistant patients, studied in a double-blind treatment protocol, demonstrated clinically significant antipsychotic responses to adjunctive alprazolam. We also briefly describe long-term efficacy of alprazolam in several patients whom we have followed in open-label clinical settings. Possible predictors or biological concomitants of benzodiazepine responsivity, which may aid in delineating distinct subgroups of patients, are discussed, and recommendations for future research are presented.


The hypothalamic lateral tuberal nucleus: normal anatomy and changes in neurological diseases.

The lateral tuberal nucleus is a circumscribed cell mass in the lateral posterior part of the hypothalamus, containing about 60000 neurons. It can be recognized in man and higher primates, probably not in other mammals. Its neurotransmitter content and connections with other parts of the brain are as yet unknown. But receptors for corticotropin-releasing factor and somatostatin, as well as muscarinic cholinergic receptors, benzodiazepine receptors and N-methyl-D-aspartate receptors have been localized within the confines of the nucleus. The lateral tuberal nucleus is affected in a number of human neurodegenerative diseases. Changes in Parkinson's disease are the least obvious: Lewy bodies appear in small amounts, the majority of them apparently lying outside a neuronal perikaryon. Neuronal loss does not occur. In Alzheimer's disease the number of neurons seems to be normal as well. Rarely silver staining tangles occur, and the deposition of A4/beta-protein in amorphous plaques is moderate. Yet, NTL neurons stain heavily in Alz-50 immunocytochemistry, while Alz-50 staining in NTL neurites is very dense. These changes are interpreted as indicating early Alzheimer-related pathology. In Huntington's disease the NTL loses neurons. This loss is related to the severity of the disease: patients who first display motor disturbances at an early age will lose more neurons than those who start later. The relation between these clinical characteristics and the severity of neuronal loss is such, that it seems likely that NTL neurons possess a special vulnerability for the effect of the Huntington gene. This could be related to their NMDA-receptor content. It is hypothesized that the NTL is involved in a neuronal network that regulates feeding and metabolism. NTL pathology may explain the peculiar catabolic state of many patients with Alzheimer's or Huntington's diseases.


The immunohistochemistry of Hodgkin's disease.

Immunohistochemistry is a valuable adjunct to the identification of Hodgkin's disease (HD) Reed-Sternberg (RS) cells, and in the differential diagnosis between HD, non-Hodgkin's lymphomas, and nonlymphoid neoplasms containing RS-like cells. The characteristic phenotype of RS cells in different subtypes of HD is described, with an emphasis on routine immunohistochemical stains. Some of the conflicting literature on this subject is reviewed to highlight pitfalls and controversies in the field.


Localization of proliferating cell nuclear antigen (PCNA/Cyclin) in workshop cases of Hodgkin's disease and non-Hodgkin's lymphoma.

Proliferating cell nuclear antigen (PCNA/Cyclin) is a 36-kD protein that is present in cycling cells but not in resting cells, and therefore represents a marker of tumor proliferation. Application of anti-PCNA/Cyclin monoclonal antibodies has shown that this protein is localized to the nucleus of cycling cells, with the exception of cells in mitosis, which demonstrate faint cytoplasmic reactivity. Recently, Benjamin and Gown found that Reed-Sternberg cells and variants show nuclear and cytoplasmic staining with anti-PCNA/Cyclin antibody 19A2, and suggested that this feature may be useful in distinguishing Hodgkin's disease from other tumors. This report describes the reactivity of 42 workshop cases that were stained with anti-PCNA/Cyclin antibodies 19A2 and/or PC10. Thirty-three (79%) of the 42 cases showed adequate reactivity to allow for interpretation of staining localization. In the group of reactive cases, 26 (79%) showed nuclear and cytoplasmic staining. The localization of PCNA/Cyclin was compared with the consensus diagnosis in each case. Eighty percent of cases classified as Hodgkin's disease, 67% of cases classified as non-Hodgkin's lymphoma, and 100% of unresolved cases showed both nuclear and cytoplasmic staining. The incidence of cytoplasmic PCNA/Cyclin was not different between Hodgkin's disease and non-Hodgkin's lymphoma in this study.


Prognostic value of kinetic parameters of HIV isolation from peripheral blood mononuclear cells.

To assess the correlations between clinical and biological stages of HIV infection and HIV isolation, peripheral blood mononuclear cells from 389 HIV-infected patients were studied by 30-day cocultures with normal lymphocytes. HIV isolation was successful in 279/389 patients (71.7%). Positive isolation was more frequent in CDC IV cases (82%) than in CDC II and III cases (63.6% and 78.4% respectively). There was a close correlation between culture positivity and serum beta 2-microglobulin, CD4+ cell counts and serum p24 antigen. The day of peak detection of reverse transcriptase activity or peak p24 antigen in coculture supernatants was selected as a coculture kinetic parameter. The day of peak detection of HIV in culture occurred earlier in CDC IV cases than in CDC II and III cases, and was a prognostic factor in AIDS progression at 2 years. These data suggest that in vitro parameters related to both viral burden and replicative capacity of HIV isolates are relevant indicators of disease progression.


Blood and urinary cadmium levels in Inuit living in Kuujjuaq, Canada.

Blood and urine cadmium concentrations have been determined in a group of 85 Inuit residents of Kuujjuaq, Quebec, Canada, drawn from actively hunting households. Mean blood cadmium values are high at 39.4 nmol/l, varying between 6.6 in non-smokers and 60.3 in smokers. No association of blood cadmium with self-reported offal consumption could be found. Median urine cadmium concentrations are elevated at 2.3 mumol/mol creatinine and rise substantially with age: 0.9 in the 30-39 age group; 3.2 among the 40-59 age group; and 4.1 in the 60 and over.


A review of the role of anti-opioid peptides in morphine tolerance and dependence.

Studies on the mechanisms of tolerance and dependence have mostly focused on changes at the receptor level. These experiments, conducted with model systems ranging from clonal cell lines to whole animals, have identified a number of important adaptive mechanisms which occur at the receptor level. However, none of these adaptive mechanisms can completely account for the phenomena which serve to define the state of morphine tolerance and dependence, especially the observation that as an animal becomes more tolerant to morphine, less naloxone is required to trigger withdrawal. The data reviewed in this paper provide strong support for the hypothesis that the brain synthesizes and secretes neuropeptides which act as part of a homeostatic system to attenuate the effects of morphine and endogenous opioid peptides. According to this model, administration of morphine releases anti-opioid peptides (AOP), which then attenuate the effects of morphine. As more morphine is given, more AOP are released, thereby producing tolerance to the effects of morphine. Cessation of morphine administration, or administration of naloxone, produces a relative excess of anti-opioid, which is in part responsible for the withdrawal syndrome. Since endogenous and exogenous antagonists might together produce synergistic effects, less naloxone might be required to trigger withdrawal in the presence of higher levels of AOPs. Although the study of AOP is in its infancy, a deeper understanding of the central nervous system (CNS) anti-opioid systems may lead to new treatments for chronic pain, substance abuse, and psychiatric disorders.


Comparison of chronic intermittent haloperidol and raclopride effects on striatal dopamine release and synaptic ultrastructure in rats.

The effects of chronic intermittent administration (7 months) of two neuroleptics, haloperidol (HAL) and raclopride (RAC), were compared using several different measures. Both drugs were administered weekly by subcutaneous injection at 7.0 mg/kg. Both neuroleptics consistently produced catalepsy throughout the treatment period, although HAL was generally more cataleptogenic than RAC. Assessment of dopamine (DA) release in the caudate putamen (CPu), through the use of in vivo microdialysis, showed that chronic HAL or RAC administration caused a prolonged decrease of DA release in response to a low dose of the DA D2 agonist quinpirole (0.03 mg/kg, sc). Injection of the muscarinic agonist pilocarpine (1.0 mg/kg, IP) did not have any significant within-group effects, although both neuroleptic treatment groups showed decreased DA release when compared to controls. Ultrastructural analysis of the dorsolateral CPu showed that both HAL and RAC treatment resulted in a significant increase in the number of perforated synapses, which contain a discontinuous density along the postsynaptic membrane. These results demonstrate that two different DA D2 receptor antagonists produce a similar effect on DA function and ultrastructural changes within the CPu following chronic, intermittent treatment.


The paradoxical effect of NMDA receptor stimulation on electrical activity of the sensorimotor cortex in freely behaving rats: analysis by combined EEG-intracerebral microdialysis.

This study was designed to determine the effects of N-methyl-D-aspartate (NMDA) receptor stimulation on the electrical activity of neocortex in freely behaving rats. Electroencephalogram (EEG) recording and intracerebral microdialysis were conducted simultaneously in the same site of the sensorimotor cortex, where the basal extracellular concentrations of aspartate and glutamate were 2.1 +/- 0.7 microM and 11.5 +/- 2.4 microM, respectively. Microdialysis with NMDA solutions (ranging from 10.0 microM to 10.0 mM) reduced the amplitude of the EEG activity and decreased the power of all frequency bands, with a virtual elimination of the high frequency waves, in a dose-dependent manner. These EEG changes were reversed after washing out the drug from the microdialysis fluid, and could be effectively antagonized with the competitive NMDA receptor antagonist DL-2-amino-5-phosphonovalerate. Remarkably, the NMDA actions were not associated with epileptiform behavioral or electrographic events. Control studies demonstrated that in the same experimental conditions, cholinergic receptor agonist carbachol caused seizures, and microdialysis with NMDA in the hippocampus readily induced epileptiform spikes. Our study shows that NMDA receptor stimulation in the rat sensorimotor cortex, although excitatory at synaptic level, can depress the local EEG activity. This may indicate that the NMDA receptor-mediated signals are processed by the neocortical network in a different way than by many other brain circuitries including hippocampus.


Fischer and Lewis rat strains differ in basal levels of neurofilament proteins and their regulation by chronic morphine in the mesolimbic dopamine system.

We studied levels of neurofilament (NF) proteins in the ventral tegmental area (VTA), and other regions of the central nervous system, of two genetically inbred rat strains, Lewis (LEW) and Fischer (F344) rats. These strains represent genetically divergent populations of rats that have been used to study possible genetic factors involved in a variety of biological processes, including drug addiction: compared to F344 rats, LEW rats show a much higher preference for several classes of drugs of abuse. We found 30-50% lower levels of three NF proteins, NF-200 (NF-H), NF-160 (NF-M), and NF-68 (NF-L), in the VTA of LEW compared to F344 rats by use of immunolabeling and Coomassie blue staining. These strain differences were highly specific to this brain region, with no differences observed elsewhere in brain or spinal cord. Interestingly, chronic treatment of F344 rats with morphine decreased levels of these three NF proteins in the VTA, as found previously in outbred Sprague-Dawley rats (Beitner-Johnson, D., Guitart, X., and Nestler, E.J.:J. Neurosci., 12:2165-2176, 1992), whereas morphine had no effect on NF levels in the VTA of LEW rats. A similar strain difference was observed in chronic morphine regulation of tyrosine hydroxylase, with morphine increasing enzyme immunoreactivity in the VTA of F344 rats (as has been observed previously in Sprague-Dawley rats [Beitner-Johnson, D., and Nestler, E.J.:J. Neurochem., 57:344-347, 1991]), but not in LEW rats. In view of the observations that LEW and F344 rats show different levels of preference for several types of drugs of abuse, and of the evidence supporting a central role of the mesolimbic dopamine system in drug reward mechanisms, the results of the current study suggest the possibility that levels of NFs and tyrosine hydroxylase may mediate some aspects of drug reinforcement and contribute to individual genetic differences in vulnerability to drug addiction.


HLA class I nucleotide sequences, 1992.

The HLA Class I sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1991 (1), Nomenclature for factors of the HLA system, 1990 (2), and Nomenclature for factors of the HLA system, 1989 (3). Due to the increased number of sequences, we have only included sequences for exons 2, 3 and 4 in this compilation. Where discrepancies have arisen between reported sequences, the original authors have been contacted where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation. In the sequence alignments, identity between nucleotides is indicated by a hyphen (-). An unavailable sequence is indicated by a period (.). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.


HLA class II nucleotide sequences, 1992.

The HLA Class II sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1991 (1), Nomenclature for factors of the HLA system, 1990 (2), and Nomenclature for factors of the HLA system, 1989 (3). Where discrepancies have arisen between reported sequences, the original authors have been contacted, where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation. In the sequence alignments, identity between residues is indicated by a hyphen (-). An unavailable sequence is indicated by an asterisk (*), gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.


[Studies of cerebrospinal fluid diagnosis in Alzheimer's disease: a review of the literature and of current developments].

Studies into the value of examination of the cerebrospinal fluid (CSF) in diagnosing Alzheimer's disease are reviewed. The diagnostic utility of about 60 substances including CSF measures related to classical neurotransmitters, (neuro)peptides, proteins, amino-acids, trace elements, and constituents of senile plaques and neurofibrillary tangles is evaluated. Technical, methodological, and ethical issues relevant to this kind of studies are discussed. None of the CSF constituents studies so far, has a proven diagnostic utility. Increased knowledge concerning macromolecular changes in the brain and improved immunochemical techniques make the outlook for a diagnostic test for Alzheimer's disease on CSF promising.


[The determination of benzodiazepines in biological objects on the Milichrome microcolumn liquid chromatograph].

1,4-benzodiazepines and benzophenols, the products of their hydrolytic cleavage, were measured by high-pressure liquid chromatography in expert material (stomach, liver, intestine, kidney samples). Milichrome microcolumn liquid chromatograph, designs 1 and 2, was employed, with a chromatography column packed with Separon C18 (5 microns) sorbent. A mixture of (NH4)2HPO4 (0.05 M) and acetonitryl in volume ratio 65:35 was used as the mobile phase in native compound analysis and that in volume ratio 45:55 for benzophenols and medazepam separation. The external standard method was used for quantitative estimation.


Analysis of T lymphocyte subsets proliferating in response to infective and UV-inactivated African swine fever viruses.

The proliferative response to infective and UV-inactivated African swine fever virus was analyzed in cells from pigs surviving an experimental infection with attenuated virus. All the pigs showed strong dose-dependent proliferative responses to both infective and UV-inactivated virus. This response was also observed when nitrocellulose-bound solubilized virus proteins were used in the assay. Heterologous isolates also induced proliferation, however it was significantly lower than that induced by the isolate used to infect the animals. The response to infective virus was blocked equally by anti-CD4 and anti-CD8 monoclonal antibodies (mAb); the response to UV-inactivated virus was almost abolished by anti-CD4 and 60% inhibited by anti-CD8 mAb. FACS analysis of 28-day T cell lines derived from peripheral blood mononuclear cells demonstrated the progressive increase of the CD8+ subset when the cells were stimulated with infective virus, whereas the stimulation with UV-inactivated virus induced the increase of both CD4+ and CD8+ subsets. In this case, the sum of CD4+ and CD8+ percentages was higher than the total percentage of T cells, suggesting the presence of cells positive for both CD4+ and CD8+.


[Self-care in the prevention of sudden cardiac death].

In acute myocardial infarction a third up to half of death registered within the first month occur in the first hour of the onset of attack most frequently because of ventricular fibrillation. Immediate self administration of drugs stabilizing electrically the heart may prevent it. On the basis of experiments in dogs and in rats flunitrazepam (Rohypnol tabl. 1 mg), tramadol (Tramal caps. 50 mg) and the beta blocker metipranolol (Trimepranol tabl. 10 mg) were selected for clinical trial on high risk patients. As the chosen combinations of drugs were not yet tested in view of possible interactions, we studied their effect on circulation and cardiovascular reflexes in eight healthy volunteers. When the drugs were absorbed from the mouth mucosa, the decrease in the heart rate during deep breathing was observed already 15 minutes after the intake of drugs. The subjects started to feel relaxed; later on they had pleasant feelings and felt sleepy. There were no undesirable changes in the heart rate or blood pressure. In the three drug combination with metripranolol, the decrease in the heart rate was more marked. The tests in volunteers were without any undesirable effects and both combinations may, therefore, be given to selected high risk subjects, e g. convalescents from myocardial infarction. Randomized trial to prove the preventive effect already started.


[Multiple endocrine neoplasia type 2: familial variant of medullary carcinoma of the thyroid gland].

In addition to a brief characteristic of the syndrome of multiple endocrine neoplasia type 2 and medullary thyroid carcinoma with a familial incidence which is a prerequisite of the syndrome, the authors submit an account on a group of 53 patients who were on the authors' records during the past 12 years. During this period the disease is systematically searched for in the families of newly diagnosed patients by examining the immunoreactive calcitonin level of relatives. Familial variants account for 28% of all medullary thyroid carcinomas. Patients who are on the records so far belong to 24 families. Approximately twice as often an isolated variant of the familial type of medullary carcinoma is involved, as compared with association with another endocrine affection, in particular pheochromocytoma (Sipple's syndrome), but associated forms will increase in number perspectively (multiple endocrine neoplasia 2A). The syndrome of multiple endocrine neoplasia 2B is very malignant but in view of the typical phenotype the disease should be diagnosed already before the change to malignancy--once the disease develops into the clinical stage the course is very adverse. From the original number of all familial tumours 38 subjects survive (72%), incl 22 who were subjected to bilateral total thyroidectomy based on screening in the preclinical stage. The prognosis of these individuals is very favourable, the calcitonin levels are throughout the follow-up period (2-10 years) repeatedly negative. With regard to the possible association with another endocrinopathy (pheochromocytoma or hyperparathyroidism) all must be followed up systematically (screening) with regard to the manifestation of an associated endocrinopathy frequently only after a longer time interval.


[Comparative evaluation of the hypotensive effectiveness of beta blockers].

The hypotensive efficacies of timolol, vistagan, and betoptic were studied in three groups of patients with primary open-angle glaucoma, matched for the clinical diagnosis, somatic condition, and number of patients. The hypotensive activity of timolol was found superior to that of vistagan or betoptic, but its administration was associated with tachiphylaxia, that is an essential disadvantage of the drug. The advantage of vistagan and betoptic is the possibility of their administration to cardiopulmonary patients with contraindications against timolol therapy.


[Vibrio cholerae toxin B subunit gene expressed in a Salmonella vaccine strain].

This paper reports that the V. cholerae toxin B subunit (ctx B) gene was inserted into pYA 248 plasmid with the aspartate beta-semialdehyde dehydrogenase (asd)gene and the recombinant plasmid was transformed into S. typhimurium deleting asd gene. Results showed that ctx B gene was highly expressed and secreted into midium. This strain was able to colonize in the intestinal epithelium. Oral immunity and general immunity could produce antibodies at high level and enhance cellular immune responses. The animals orally inoculated with S. typhimurium x 4072 (pYA-ctx B) vaccine had remarkable protection against virulent V. cholear 569B strain and S. typhimurium strain. Use of such system provides useful method for oral vaccine.


[Carcinoma in situ in cryptorchid testes in post-pubertal patients].

The pathohistologic analysis of testis sections of 37 postpuberty patients with different types of cryptorchism is performed. The tissue samples were taken during orchiopehy, fixed in Bouin's solution and treated by the standard histologic techniques. The morphologic criteria are presented for identification of the presence of the so called carcinoma in situ cells found in two cases. Besides, in 13 patients rare, mainly single, atypical germinative cells were found in a smaller number of the seminiferous tubules. It has been concluded that the presence of carcinoma in situ cells in undescended testes of some patients and considering the simple way of sampling, lack of complications and high reliability of the diagnostic procedure, it is absolutely justified to take routine biopsy of testes during orchiopexy in each postpuberty and perhaps prepuberty patient.


[The effect of fenazepam on the humoral immune response in secondary immunodeficiency].

A single administration of phenazepam (2.5 mg/kg) enhances the synthesis of antibodies after immunization with different vaccines. Phenazepam restores antibody formation in immunodeficiency induced by intoxication. The immunostimulating effect of phenazepam is linked with an increase in the capacity of macrophages for inducing humoral immune response and a rise in the number of antibody-producing cells in the spleen.


Limbic seizure-induced changes in extracellular amino acid levels in the hippocampal formation: a microdialysis study of freely moving rats.

Limbic seizure-activity was induced by injecting kainic acid into the amygdala of rats. Extracellular levels of amino acids were monitored by microdialysis in the hippocampus. No changes were detected in the levels of glutamate and aspartate. The level of glycine also remained unchanged, whereas GABA showed an increase of approximately 35%. The level of glutamine decreased by approximately 30%, and that of serine by approximately 20%. The results indicate that increased turnover may exist in the glutamate transmitter pool. In addition, impairment of GABA-release seems not to be a pathogenetic factor in seizure-induced hippocampal neuron loss. It is concluded that even during sustained seizure-activity, the extracellular level of glutamate, is maintained within narrow limits. A proposed index for excitatory neurodegeneration, glutamate x glycine/GABA, was found to be decreased in this seizure model. We therefore suggest that seizure-induced neuron death is not reflected by alterations in the extracellular levels of glutamate and aspartate, thought to act as direct neurotoxins.


Glutamate and aspartate are decreased in the skin in amyotrophic lateral sclerosis.

We measured the levels of amino acids in biopsied skin from eight patients with amyotrophic lateral sclerosis (ALS) and seven controls. The most conspicuous changes in ALS patients were as follows. First, the contents of the acidic amino acids glutamate and aspartate were significantly decreased in ALS, and were negatively and significantly associated with the duration of illness. Second, the levels of the collagen-associated amino acids hydroxyproline, proline, glycine, alanine, and hydroxylysine were significantly decreased in ALS, and correlated inversely with the duration of illness. These results suggest that there are abnormalities of acidic amino acids and collagen-associated amino acids in the skin of patients with ALS. These changes may underlie the pathogenesis of ALS.


[Amphetamine--induced rage reaction in mice and its mechanism].

Rage reaction was induced in mice by ip amphetamine sulfate (APT) 15 mg/kg. Mice appeared hyperreactive after 6 min and then squeaked and fought each other. These manifestations were most distinct in 15-30 min and subsided after 40-70 min. At 20 degrees C and 25 degrees C, the occurrence of rage reaction was 85.0% and 90.0% respectively. The ED50 of APT for eliciting rage reaction was 11.8 +/- 2.1 mg/kg ip. No significant difference in the induction of rage reaction was observed between male and female mice but ambient temperature affected the occurrence of this reaction. Neuroleptic drugs (chlorpromazine, haloperidol, tardan and clozapine), anxiolytic drugs (diazepam and meprobamate) and reserpine suppressed the rage reaction induced by APT in mice. Phenobarbital and pentobarbital (at sedative doses), atropine, scopolamine, phentolamine and propranolol exerted no influence on APT--induced rage reaction. Amantadine, levodopa and apomorphine at lower doses potentiated the rage inducing effect of APT. Moreover, at higher doses amantadine or levodopa alone also evoked rage reaction similar to that induced by APT. Therefore, it may be deduced that the APT-induced rage reaction results from increased release of dopamine in limbic system and has nothing to do with the simultaneous epinephrine release. The available data indicate that the APT--induced rage reaction in mice deserves to be recommended as an animal model for screening potential neuroleptic drugs. The merits and shortcomings of this new model are discussed.


Vagally mediated release of gastrin and cholecystokinin following sensory stimulation.

The aim of the present study was to investigate whether gastrin, cholecystokinin (CCK) and somatostatin secretion can be influenced by sensory stimulation and if so, whether such effects are mediated via the vagal nerves. Male rats anaesthetized with chloral hydrate were exposed to three different stimuli, i.e. to low frequency (2 Hz) electrical stimulation of muscles via needles (electro-acupuncture), to thermal stimulation at 40 degrees C or to vibration at 100 Hz. The two former stimuli activate mainly small and medium sized myelinated fibres from muscles and skin respectively, whereas vibration activates large myelinated fibres from skin, subcutaneous tissue and muscles. Experiments were also performed on animals that were vagotomized or exposed to prior treatment with atropine (0.5 mg kg-1). Blood was collected at various time intervals and plasma levels of gastrin, CCK and somatostatin were measured with radioimmunoassay (RIA). All three stimuli, i.e. electro-acupuncture, vibration and thermal stimulation caused significant elevations of gastrin (103 +/- 11-151 +/- 16 pM, 105 +/- 8-140 +/- 12 pM and 105 +/- 14-162 +/- 4 pM) and cholecystokinin (9 +/- 0.8-15 +/- 2.8 pM, 8 +/- 0.5-10 +/- 1.5 pM and 8.0 +/- 0.5-10.5 +/- 1.5). Somatostatin was raised in response to electro-acupuncture (10 +/- 1-14 +/- 3 pM). Vagotomy and atropinization abolished the release of gastrin and CCK in response to all three stimuli. CCK levels were significantly reduced following electro-acupuncture in atropinized rats. In conclusion, gastrin and cholecystokinin release is stimulated by activation of sensory afferent, originating in skin, subcutaneous tissue as well as in muscle.(ABSTRACT TRUNCATED AT 250 WORDS)


Intracranial haemodynamics in Takayasu's arteritis.

The cerebral circulation was assessed in two cases of Takayasu's arteritis by angiography of the aortic arch, cerebral blood flow single photon emission computed tomography (SPECT) and transcranial Doppler sonography (TCD). In both cases, disease of the major brachiocephalic arteries affected flow in the vertebrobasilar system and circle of Willis. Basilar artery flow was permanently reversed in one case, with vertebrovertebral shunting and subclavian steal. In the other case, focally reduced cerebral hemisphere flow resulted from watershed between vascular territories. The complex collateral and haemodynamic changes produced by multi-vessel involvement in Takayasu's arteritis suggest that therapeutic approaches should be based on assessment of end-organ perfusion rather than on structural angiographic changes.


Hepatitis B vaccine alone or in combination with anti-HBs immunoglobulin in the perinatal prophylaxis of babies born to HBsAg carrier mothers.

The efficacy of hepatitis B virus (HBV) vaccine alone (group I) or in combination with hepatitis B immunoglobulin (HBIG) (group II) for prevention of perinatal transmission of the virus was assessed in 21 and 24 neonates, respectively. 58 infants who could not be vaccinated constituted the control group. It was observed that in the unvaccinated group approximately 70% of the infants became infected. In both the vaccinated groups, the seroconversion and seroprotection rates (anti-HBs > or = 10 IU/1) were almost similar at 6 months of follow up, but, at 12 months, infants given HBIG and vaccine showed better seroprotection rate (85%) than those given vaccine alone (58.8%). Immune response to the vaccine was also better in both the groups if the mothers were anti-HBe positive. Despite immunization, 14.2% and 25% infants in group I and II, respectively, became chronic carriers if their mothers were HBeAG positive.


Southern blot hybridization analysis of polyoma virus-specific RNA synthesized under the block of virus replication by 5-bromo-2'-deoxyuridine.

Polyoma (Py) virus-specific RNA, synthesized at reduced level in infected cells in the presence of antiviral substance 5-bromo-2'-deoxyuridine was characterized in more detail by Southern blot hybridization analysis. Virus-specific RNA present in 3H-uridine labelled cytoplasmic preparations hybridized to individual viral DNA restriction fragments in a characteristic manner and extent, which was quantitatively assayed. BrdUrd (6.34 micrograms/ml) lowers the hybridization profiles essentially in a proportional manner, which indicates that no new, atypical transcription products are formed in the presence of BrdUrd.


Induction of interferon synthesis and cytotoxicity by murine peritoneal macrophages exposed to glycoprotein ligands.

Thioglycollate-induced murine C57BL/6 and C3H/HeN peritoneal macrophages synthesized interferon-beta (IFN-beta) in response to exposure to glycoproteins such as horseradish peroxidase (HRP) or mannosyl or fucosyl bovine serum albumin (BSAman of BSAfuc, respectively), but not glucosylated or galactosylated BSA (BSAglu or BSAgal, respectively). These results suggest participation of the mannosyl-fucosyl receptor (MFR) in this response. IFN synthesis was augmented by culturing macrophages in L cell-conditioned medium prior to exposure to these substances. Macrophages obtained from lipopolysaccharide (LPS)-resistant C3H/HeJ mice did not produce IFN in response to HRP. Furthermore, IFN-induction by HRP was blocked by polymyxin B. In addition, exposure of macrophages to HRP or BSAman induced cytotoxicity against NIH 3T12 cells. Cytotoxicity was not inhibited by the presence of anti-IFN-alpha/beta. In contrast to IFN induction, however, macrophages activation was LPS-independent, since this activity was demonstrated in macrophages from C3H/Hej mice. The carbohydrate specificity of these responses suggests that the MFR or an another scavenger receptor may be involved in the responses to these substances, and that cytotoxicity and IFN-induction by glycoproteins follow unique pathways.


Prevalence and specificity of lymphocytotoxic antibodies in different stages of HIV infection.

Sera obtained from 27 HIV-infected persons were investigated for complement-dependent humoral cytotoxicity. Uninfected as well as HTLV-IIIB-infected H9 cells were used as cellular targets either before or after stimulation by phytohemagglutinin (PHA) or concanavalin A (Con-A). The degree of cytotoxicity was determined by 51Cr-release assay. Two different antibodies could be found in sera of HIV-infected persons, one being directed against HIV-induced cell surface component(s) and the other reacting with structure(s) present on activated T4 cells. Asymptomatic HIV-carries were found to have antibodies exerting complement-dependent cytotoxicity to HIV-infected T4 cells. These antibodies were reactive mainly after stimulation of HIV-infected target cells by Con-A. Sera of ARC and AIDS patients contained autoantibodies reactive with PHA-stimulated or HIV-infected T4 lymphocytes. These data suggest that HIV-specific antibodies represent an anti-viral immune defense, while autoantibodies may be important in destruction of the immune system in AIDS.


Monoclonal antibody to Japanese encephalitis virus cross-reacting with histones present in the cell nuclei.

An immunoglobulin G (IgG2b) class of monoclonal antibody (MoAb, NHA-1) raised against Japanese encephalitis virus (JEV) E glycoprotein, reacted with the viral antigen expressed in cytoplasm of the infected cells and also with the cell nuclei, by an indirect fluorescent antibody technique (FA). The NHA-1 reactivity to nuclei was found to be due to its recognizing a JEV cross-reactive epitope present on the nuclear histones. Adsorption with calf thymus histones (type II-AS) showed a drop in NHA-1 reactivity to both JEV and histones by an enzyme-linked immunosorbent assay (ELISA) and indirect FA; the drop was higher against the histones. The MoAb recognized specifically the viral antigens expressed on the infected porcine kidney cell surface by a modified indirect FA. ELISA carried out with glutaraldehyde-fixed antigens showed an almost 2-fold increase in the reactivity over unfixed JEV antigen but none for the histones. Thus, the results indicate that histones share a sequential homology with E glycoprotein of JEV, which might lead to an autoimmune disorder induced due to the molecular mimicry between these two antigens.


Experimental studies on the susceptibility of Aedes vittatus to dengue viruses.

Ae. vittatus mosquitoes were infected by oral route and by intrathoracic inoculation with dengue (DEN) viruses and tested for the presence of dengue virus antigen in their head squashes and salivary glands by indirect immunofluorescence. The results indicate that this species was susceptible to all four types of DEN viruses and supported the growth of DEN-2 virus.


Prenatal exclusion of choroideremia.

We performed prenatal testing to predict the inheritance of choroideremia (CHM) using a linked polymorphic DNA marker, DXS95. DNA analysis of chorionic villi at the 12th week of pregnancy indicated that the allele at risk had not been passed from the heterozygous mother to the fetus. This prenatal exclusion of choroideremia was confirmed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis.


Renal dopamine receptors and pre- and post-cAMP-mediated Na+ transport defect in spontaneously hypertensive rats.

We have reported defective coupling of the renal tubular DA1 dopamine receptor to adenylyl cyclase in both the spontaneously hypertensive rat (SHR) and the Dahl salt-sensitive rat. Since Na+, 5'-guanyl imidodiphosphate [Gpp(NH)p], and N-ethylmaleimide (NEM) reduce agonist affinity for brain D1 dopamine receptors, we compared the effects of these agents on agonist affinity in proximal tubules from SHR and its normotensive control, the Wistar-Kyoto rat (WKY), to delineate further the site of the DA1-adenylyl cyclase coupling defect. In WKY, the D1/DA1 agonist, fenoldopam, competed for 125I-Sch 23982 at a high-affinity site (KiH = 1.8 +/- 0.8 x 10(-8) M) and a low-affinity site (KiL = 7.6 +/- 1.1 x 10(-5) M, n = 6). Na+ (150 mM) or Gpp(NH)p (10(-4) M) converted KiH to KiL. NEM, which alkylates sulfhydryl groups, also converted all the binding to KiL; this effect could be prevented by prior treatment with 10(-4) M fenoldopam. In contrast, in SHR, fenoldopam detected only a KiL (7.8 +/- 1.4 x 10(-5) M, n = 6). Neither Na+, Gpp(NH)p, nor NEM had any effect on KiL. To study a functional expression of these binding sites, the effect of 5 x 10(-5) M fenoldopam or 8-(chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (8-CPT-cAMP) on Na+/H+ exchange activity in proximal tubular brush-border membrane vesicles was tested. In WKY, the inhibitory effects of these agents on the exchanger increased with the age of the rat.(ABSTRACT TRUNCATED AT 250 WORDS)


Hormone signaling systems in inner medullary collecting ducts.

The inner medullary collecting duct is a complex tissue that exhibits a variety of hormone signaling systems. These include the following: adenylyl cyclase activity stimulated by vasopressin (AVP), beta-adrenergic agonists, or prostanoids and inhibited by alpha 2-adrenergic agents or adenosine; guanylate cyclase activity in response to atrial natriuretic peptide (ANP); phospholipase C activity stimulated by ANP, AVP, bradykinin, endothelin, epidermal growth factor (EGF), and muscarinic cholinergic agents; and phospholipase A2 activity stimulated by AVP, bradykinin, EGF, and endothelin. The signal transduction mechanisms for each of these hormone signaling systems is succinctly reviewed, and the interactions between different signaling pathways are discussed. Central to this interaction is the mutually inhibitory relationship between activation of adenylyl cyclase and phospholipases. Increasing cellular adenosine 3',5'-cyclic monophosphate content impairs activation of phospholipases A2 and C; conversely, stimulation of phospholipase C impairs AVP-stimulated adenylyl cyclase activity via activation of protein kinase C.


Phalloidin prevents leukocyte emigration induced by proinflammatory stimuli in rat mesentery.

The objective of this study was to determine whether phalloidin, a potent microfilament stabilizer, can modify inflammatory mediator-induced leukocyte adhesion and extravasation in postcapillary venules of the rat mesentery. To address this issue, the rat mesentery was prepared for in vivo microscopic observation. Venules with initial diameters ranging between 25 and 35 microns were selected for study. Erythrocyte velocity, vessel diameter, leukocyte rolling velocity, and the number of adherent (stationary for 30 s) and emigrated leukocytes were initially determined during superfusion of the mesentery with phosphate-buffered saline. After these variables were recorded during the control period, either 100 nM platelet-activating factor (PAF), 20 nM leukotriene B4 (LTB4), or 1 microM N-formyl-methionyl-leucyl-phenylalanine (FMLP) was added to the superfusate. Repeat measurements were obtained between 50 and 60 min after initial exposure to the inflammatory mediator. In some experiments, rats were given phalloidin (25 or 500 micrograms/kg iv) 30 min before superfusion with the inflammatory mediators. Superfusion of the mesentery with either PAF, LTB4, or FMLP enhanced leukocyte adherence and emigration and reduced leukocyte rolling velocity. Pretreatment with the low dose of phalloidin effectively prevented leukocyte emigration but had no effect on the increased leukocyte adherence elicited by the three inflammatory mediators. However, when administered at the higher dose, phalloidin prevented both leukocyte adherence and emigration. Neither dose of phalloidin altered the upregulation of neutrophil membrane CD11/CD18 glycoprotein adherence complex induced by PAF or LTB4. These results are consistent with the concept that PAF, LTB4, and FMLP increase leukocyte extravasation by a process that may involve alterations in the endothelial cell cytoskeleton.


Alpha-adrenergic vasoconstriction in normal and hypoperfused myocardium during sympathetic nerve stimulation.

Coronary vasoconstriction mediated by postjunctional alpha 1- and alpha 2-adrenergic receptors was studied in normally perfused (control group) and left coronary hypoperfused (stenosis group) hearts of vagotomized, beta-blocked (propranolol) cats. Cardiac sympathetic nerve stimulation was combined with alpha 1- and subsequent alpha 2-adrenergic antagonism (doxazosin and SK&F 104078). Coronary perfusion pressure and heart rate were kept constant within groups; regional myocardial blood flow and cardiac output were obtained by means of microspheres with concomitant measurement of left ventricular myocardial oxygen consumption (MVO2). alpha 1-Adrenergic antagonism alone did not significantly alter blood flow in any wall layer in either group. Subsequent alpha 2-adrenergic antagonism increased epicardial as well as composite transmural flow in the stenosis group (P < 0.025). The inverse correlation between coronary resistance and MVO2 vanished in the stenosis group following alpha 1- and alpha 2-adrenergic antagonism. Maximal first derivative of the left ventricular pressure-time relation (dP/dt) and cardiac output were reduced simultaneously (P < 0.001). Hence, the significance of alpha 1- and alpha 2-adrenergic stimulation of inotropy and cardiac performance are augmented by myocardial hypoperfusion. Furthermore, alpha 2-adrenergic receptors are responsible for epicardial vasoconstriction in hypoperfused myocardium.


Effect of afterload and beta-adrenergic blockade on nonischemic myocardial contraction pattern.

We studied how changes in afterload affect regional contraction in the anterior wall of the left ventricular after circumflex coronary arterial (CFX) occlusion and subsequent beta-adrenergic blockade in pentobarbital sodium-anesthetized cats. Regional function was determined by orthogonal sonomicrometry. CFX occlusion produced nonuniform hyperkinesis in the nonischemic anterior wall; shortening of circumferential segments increased from 10.1 to 14.1% (P < 0.001), whereas shortening of longitudinal segments increased from 3.0 to 9.6% (P < 0.001). Hyperkinesis of longitudinal segments was influenced neither by changes in afterload over a pressure range of +/- 30 mmHg nor by beta-adrenergic blockade, indicating that hyperkinesis of longitudinal segments does not rely on increased inotropic state or resistance to ventricular emptying. Hyperkinesis of longitudinal segments occurred at end-diastolic lengths equal to preocclusion conditions, whereas hyperkinesis of circumferential segments was dependent on activation of the Frank-Starling mechanism. Furthermore, shortening of circumferential segments decreased with increments in afterload, particularly after CFX occlusion and subsequent beta-adrenergic blockade. In conclusion, CFX occlusion alters the contraction pattern of the nonischemic anterior wall. The postocclusion contraction is sensitive to increased afterload in the cardiac minor axis direction. These initial alterations may well direct the following remodeling process in infarcted hearts.


Baroreflex control of regional capacitance and blood flow distribution with or without alpha-adrenergic blockade.

Regional blood volumes (Vb), unstressed volumes (V0), blood flow distribution, venous compliances (Cv), venous resistances (Rv), and time constants of drainage (tau v) were determined in dogs anesthetized with alpha-chloralose at carotid sinus pressures (Pcs) of 50 and 200 mmHg and dosed with alpha-adrenergic or ganglionic blockade at a Pcs of 50 mmHg. Vb was measured in each region from indicator dilution curves and mean transit times. V0 was extrapolated from the pressure-volume curves. Pcs of 50 and 200 mmHg were maintained in random order. With a decrease in Pcs, arterial pressure increased from 58.7 +/- 4.1 to 104.6 +/- 6.4 mmHg (P < 0.01), peripheral fractional blood flow decreased from 69.8 +/- 3.8 to 55.8 +/- 3.9% (P < 0.001), splanchnic Vb decreased from 28.3 +/- 1.9 to 19.3 +/- 1.2 ml/kg (P < 0.01), and splanchnic V0 decreased from 19.6 +/- 1.4 to 6.3 +/- 2.1 ml/kg (P < 0.001). Splanchnic Rv and tau v also decreased, whereas splanchnic Cv increased. Phentolamine at low Pcs only partially reversed the decrease in splanchnic capacitance, whereas hexamethonium completely abolished it. In conclusion, changes in splanchnic Rv and blood flow distribution are important components of the carotid sinus reflex, and alpha-adrenergic receptor activation is only partially responsible for the changes in vascular capacitance by the baroreceptor reflex.


Desensitization to acetylcholine in single sinoatrial node cells isolated from rabbit hearts.

The negative chronotropic effect of acetylcholine (ACh) on the sinoatrial node fades in the continuous presence of ACh as a result of desensitization. We have investigated the mechanism underlying desensitization in single rabbit sinoatrial node cells using the whole cell patch clamp technique. The negative chronotropic effect resulting from the injection of a constant hyperpolarizing current faded. ACh activated an inwardly rectifying potassium current (iK,ACh), which faded in the continuous presence of ACh. ACh had no effect on "basal" L-type calcium current (iCa), but ACh decreased iCa, which had been potentiated by isoprenaline. This effect did not fade during a 2-min exposure to ACh. ACh decreased the hyperpolarization-activated current (i(f)). This effect again did not fade. These results suggest that desensitization of the negative chronotropic response to ACh is, in part, the result of the membrane hyperpolarization and, in part, the result of the fade of iK,ACh. These results also suggest that, whereas the activation of potassium current by ACh rapidly fades, the effects resulting from the inhibition of adenylate cyclase do not.


Alpha 2-adrenoceptors mediate norepinephrine constriction of porcine pial veins.

The adrenergic innervation and alpha-adrenoceptor agonist-induced constrictions in isolated medium-size pial veins (OD 734 +/- 18 microns) of the pig were investigated. Using in vitro tissue bath techniques, we noted exogenously applied norepinephrine (10(-9) to 10(-5) M) induced venoconstriction with EC50 values of 1.71 x 10(-7) M, where EC50 is the concentration that produced 50% of (non-KCl) agonist-induced maximum constriction. The constriction was mimicked by clonidine but not by phenylephrine and was more effectively blocked by yohimbine than by prazosin. Results from histochemical studies demonstrated that porcine pial veins received a denser plexus of catecholamine fluorescence fibers than do pial arteries with similar outer diameter. These results suggest that norepinephrine-induced pial venoconstriction is mediated predominantly by alpha 2-adrenoceptors and that porcine pial veins have significantly greater sensitivity to the alpha-action of norepinephrine than that reported in pial arteries. These results add further support for adrenergic innervation in pial veins being of importance in regulating cerebral blood volume and intracranial pressure.


Age-related decline in left ventricular filling at rest and exercise.

To determine whether the age-associated decline in resting left ventricular diastolic filling persists during aerobic exercise, rest and bicycle exercise filling indexes were measured from gated radionuclide blood pool scans in 88 healthy men aged 22-82 yr. To evaluate the effect of physical conditioning status on these age-related changes, a subset of the subjects consisted of endurance-trained senior athletes with a maximal O2 consumption of 50.5 +/- 5 compared with 32.6 +/- 7 ml.kg-1 x min-1 in age-matched controls. The contribution of beta-adrenergic stimulation to exercise-induced changes in filling was also evaluated by the administration of intravenous propranolol to another subset before testing. Peak filling rate increased progressively at all ages with increasing exercise work loads. The peak filling rates at rest, 50% maximal exercise, and maximum exercise inversely correlated with age (r = -0.64, -0.53, -0.64, respectively). Rest and exercise filling indexes in senior athletes were similar to those of sedentary older subjects. Propranolol decreased exercise peak filling rates in young (37.2 +/- 7.5 yr) but not in older (62.1 +/- 6 yr) subjects. Therefore, filling rates increase with exercise in both young and older healthy men, but age differences persist at comparable relative work loads. This decline is not secondary to a decline in physical conditioning status but appears to be related to a decrease in beta-adrenergic responsiveness in older individuals.


[Undescended testicle treated with FSH and HCG].

Medical treatment with FSH and HCG was tested during 21 days in 47 cases (76 testis) of undescended testis. The average testis size, as measured by ecography, increased from 0.828 cc to 1.57 cc 48 hours after completing the treatment. Three months later the average testis size was 1.025 cc and 1.24 cc at six months. FSH increased slightly and total testosterone and free testosterone increased significantly. After treatment, 68% of the previously undescended testis descended.


[Non-Hodgkin's malignant lymphoma and human immunodeficiency virus. Apropos of 34 cases].

The characteristics of 34 HIV-associated non Hodgkin's lymphomas diagnosed and treated at Bordeaux hospitals are described. The patients represented 7% of the AIDS cases observed in the Bordeaux area. HIV-lymphomas were almost always high-grade malignancies, usually disseminated (70%) with extranodal disease at presentation (91%) primarily in the bone marrow, meninges, gastrointestinal tract and liver. Twenty-eight patients were treated with different chemotherapy protocols or radiation therapy alone. Complete remission was achieved in 11 patients and partial remission in 3. The median survival was 3.9 months. Despite utilization of low-intensity chemotherapy regimens, opportunistic infections were not prevented. The only factor that accurately predicted complete remission was the WHO performance index. The total number of CD4-positive lymphocytes, the Ann Arbor stage and the WHO performance index were prognostic factors influencing survival. These results justify the use of high-intensity regimens, but only for patients without opportunistic infection and with a WHO performance index below 3.


[Esophageal candidiasis in HIV positive patients. Lack of statistical correlation with CD4 lymphocytes].

One hundred and twenty-nine upper gastrointestinal endoscopies were performed between November, 1986, and November, 1990, at the Fort-de-France Hospital (Martinique), on 92 patients who were seropositive for the human immunodeficiency virus (HIV). Blood samples were drawn at the same time as 75 endoscopic examinations and the number of CD4 lymphocytes/mm3 was determined. The incidence of esophageal candidiasis was 23.1% and lesions were observed in 17.2% of the endoscopies performed systematically at the time that the patient's seropositivity was first established. No significant relationship was found between the presence of esophageal candidiasis and peripheral blood CD4 lymphocyte counts.


Survey of the methicillin resistance-associated genes mecA, mecR1-mecI, and femA-femB in clinical isolates of methicillin-resistant Staphylococcus aureus.

The restriction site polymorphism of the chromosomal femAB region and the first appearance of the regulatory element mecR1-mecI associated with the methicillin resistance determinant (mec) were analyzed in 192 initially methicillin resistant (Mcr) Staphylococcus aureus clinical isolates collected between 1965 and 1990 in the Zurich area. Forty-three of the strains lost the resistance spontaneously. All isolates that were still Mcr hybridized with mecA, the gene for the low-affinity penicillin-binding protein PBP 2'. Mcr strains isolated before 1977 lacked sequences that hybridized with mecR1-mecI, a regulatory element controlling the expression of mecA; exceptions to this were one strain isolated in 1966 and one strain isolated in 1972. The size of the EcoRV fragment carrying femA, a chromosomally encoded factor involved in pentaglycine side chain formation of the peptidoglycan and essential for the expression of methicillin resistance, was conserved in all strains but one, which was susceptible to methicillin even though it carried a functional mecA gene. The methicillin susceptibility of this particular strain was presumably due to a spontaneous femA-like mutation. The 192 strains belonged to seven different EcoRV restriction fragment patterns recognizable with a 10.5-kb probe covering the femAB region. Some 93% of the 149 Mcr strains belonged to pattern A, and the remaining Mcr strains shared patterns A' and B. The 42 isolates which spontaneously lost their resistance upon storage and revival represented all seven different patterns. This strong conservation of femA suggests an important role for femA in cell wall metabolism and methicillin resistance.


[Is there any desensitization of presynaptic alpha 2-adrenergic receptors in hypertension? Experimental and clinical studies].

Several authors have discussed an alteration of adrenergic receptivity in arterial hypertension. De Champlain (Hypertension 1990; 8: S77-S85) suggested that postsynaptic alpha 1-adrenergic functions became dominant while beta-adrenergic functions are attenuated in arterial hypertension. However, the status of presynaptic alpha 2-adrenoceptors remains unknown. The present study investigates presynaptic alpha 2-adrenoceptors in hypertension through the measurement of plasma levels of noradrenaline after administration of yohimbine, an alpha 2-adrenoceptor antagonist, in essential hypertension. Yohimbine (0.2 mg/kg per os) induced a 73% increase of plasma levels of noradrenaline in hypertensive patients (n = 12) and a 178% one in normotensive subjects (n = 6, p < 0.05). A similar significant difference was found in experimental neurogenic hypertension observed in awake dogs 3 weeks after sinoaortic denervation: the increase in plasma concentrations of noradrenaline after yohimbine (0.5 mg/kg i.v.) was +279% in hypertensive versus +642% in normotensive dogs (p < 0.05). The results show that the magnitude of the yohimbine-induced sympathetic activation is lower in hypertensives than in normotensives. They suggest the existence of a presynaptic alpha 2-adrenoceptor desensitization in arterial hypertension. The abnormality of this presynaptic inhibitory mechanism can increase the sympathetic tone and help to develop and maintain arterial hypertension.


[Soluble intercellular adhesion molecule-I levels in sera of patients with Kawasaki disease].

We investigated whether serum levels of soluble ICAM-1 antigens increases during acute Kawasaki disease (KD). We also compared levels in acute KD with those in anaphylactoid purpura (AP) and in measles. Serum soluble ICAM-1 levels were measured by a double determinant immunoassay using two monoclonal antibodies in the FAST system. Serum levels of tumor necrosis factor-alpha (TNF-alpha) were measured by a specific and sensitive sandwich enzyme immunoassay. Patients with KD, but not with AP or measles, had increased ICAM-1 levels in serum during the acute stage. In addition, during the acute stage, KD patients with coronary-artery lesions (CAL) were found to have increased ICAM-1 levels in serum compared to patients without CAL. We found a positive correlation between serum levels of ICAM-1 and levels of TNF-alpha during acute KD. Our results suggest that the serum level of soluble ICAM-1 is an important immunologic parameter for determining the severity of vascular damage during acute KD.


The expression of presynaptic t-ACPD receptor in rat cerebellum.

The expression of a receptor subtype for one type of excitatory amino acid agonist, t-ACPD, was examined in developing Purkinje cells of cerebellar slices. The t-ACPD-induced responses were compared with those induced by QA in current response, single cell Ca2+ imaging and changes in the miniature currents in the same preparation. It was found that t-ACPD induced a single component of inward current, and an increase in the frequency of miniature currents associated with the presence of external Ca2+, but was ineffective at mobilizing intracellular Ca2+ even in the presence of external Ca2+. The present study suggests the expression of at least two types of metabotropic receptors in the Purkinje cell region, one of which, expressed in the Purkinje cell dendrites, is highly sensitive to QA, but relatively insensitive to t-ACPD, and the other of which is a t-ACPD-sensitive receptor expressed on the presynaptic terminals of the neurons making synapses onto Purkinje cells.


Molecular characterization of the gene encoding glutamine synthetase in the cyanobacterium Calothrix sp. PCC 7601.

In order to study the regulation of the synthesis of glutamine synthetase in response to changes in environmental parameters (light and nitrogen sources), we have cloned and sequenced the glnA gene from the filamentous cyanobacterium Calothrix PCC 7601. This gene consists of 472 codons and encodes a polypeptide of M(r) 52,290 highly homologous to that from Anabaena PCC 7120, but more distant from those identified from other procaryotes. The relative abundance of the two glnA transcripts (1.6 and 1.8 kb) is equivalent in cells grown under either red or green light, but the 1.6-kb species predominates in nitrate-grown cells and the 1.8-kb species in ammonia-grown cells. The very high identity (74%) observed between the 374-bp long nucleotide sequence upstream from the Calothrix and Anabaena glnA genes suggests the existence of similar regulatory signals for the control of glnA expression in both cyanobacteria.


Evidence of tyrosine hydroxylase mRNA in the anterior and neurointermediate lobes of female rat pituitary.

The anterior pituitary is thought to be unable to synthesize dopamine (DA) except under experimental conditions where a tyrosine hydroxylase (TH) activity, the rate-limiting step of its synthesis, has been demonstrated. In this work, we tested whether the enzyme described as active under particular conditions comes from de novo TH gene transcription or from a pre-existing TH mRNA poorly translated or untranslated under physiological conditions. Therefore, we searched for the presence of TH mRNA in normal female rat pituitary using the polymerase chain reaction following reverse transcription (RT/PCR) and in situ hybridization (ISH). The neurointermediate lobe (NIL) of the hypophysis was used as negative tissue, since it is thought to be unable to synthesize TH. As expected, no ISH labelling could be seen in the neural lobe (NL). However, scarce labelled cells were found in the intermediate lobe (IL) confirming the positive results observed in the NIL by RT/PCR. The anterior lobe (AL) also presented TH mRNA by PCR and ISH. The TH gene expression in sparse cells of the AL is discussed in regard to the ability of the AL to synthesize DA under particular conditions from a pre-existing mRNA.


[Creatine phosphokinase in a hospitalized psychiatric population].

Creatine phosphokinase (CPK) is studied in psychiatric hospitalized patients under neuroleptic treatment in order to clarify the diagnostic value of this enzyme in the neuroleptic malignant syndrome (SNM). Intramuscular drug administration was, in the present study, the only variable associated with CPK levels over 1,000 U. Other items studied, like agitation, physical restraint or illness severity can not account for this elevation. In absence of i.m. medication, levels over 1,000 U must be carefully screened in order to rule out SNM or organic pathology associated.


Uptake of broxaterol by cultured human cells.

The mechanism of uptake of the sympathomimetic drug broxaterol by Chang liver and HepG2 cell lines was investigated. When cells were incubated in the presence of low concentrations of broxaterol cell cultures take up (t1/2 = 5-10 min) and, depending on the experimental conditions, accumulate the drug such that the intracellular concentration is over 200-1000 times that in the incubation medium. The uptake was saturable and influenced by the presence of sodium and variations in external pH. These data may represent a model for tissue uptake in vivo in an attempt to investigate that selective uptake of broxaterol is involved in withdrawal of the molecule during drug therapy.


Increase in activating ability of human platelet guanylate cyclase during aggregation.

The dynamics of changes in the stimulation of human platelet guanylate cyclase by some activators in aggregating platelets was studied. It was shown that ADP-induced aggregation of human platelets (donors) is accompanied by the enhancement of the intensity of guanylate cyclase activation by sodium nitroprusside, L-arginine, protoporphyrin IX and arachidonic acid and also by the increase in cGMP content. Immediately after the induction of aggregation the intensity of guanylate cyclase activation and cGMP content begin to increase. The rise reaches its maxima within several minutes, then followed by a fall to the initial level. The peaks of the enhanced capacity for guanylate cyclase activation by the above compounds coincide in time and intensity. On the basis of the proposed hypothetical scheme of cGMP action as a regulator of platelet aggregation a possible mechanism of enhancing the capacity of guanylate cyclase to be stimulated by various activators in aggregating platelets is suggested.


omega-Dialkylaminoalkyl esters of N-phenyl aminethiocarboximidic acids with local anesthetic and other activities.

A series of omega-dialkylaminoalkyl esters of N-phenyl aminethiocarboximidic acids was prepared by reaction of N-phenyl 1-pyrrolidine, 1-piperidine, 4-morpholine, 1,2,3,4-tetrahydro-1-quinoline, 1,2,3,4-tetrahydro-2-isoquinoline, 10-phenothiazine, N-phenyl-2-pyridylamine and N-benzyl-2-pyridylamine-carbothioamides (prepared in situ from the appropriate secondary amine and phenyl isothiocyanate) with a number of omega-chloroalkyldialkylamines in anhydrous DMF or THF solution in the presence of sodium hydride. Good yields of the above esters were obtained provided that sodium hydride was added initially or after the formation of 3,3-disubstituted 1-phenylthiourea, according to the nature of the secondary amine. Some esters showed in mice local anesthetic activity comparable with that of lidocaine, as well as moderate hypoglycemic, antiarrhythmic, analgesic, antiacetylcholine, H1-antithistamine and platelet antiaggregating activities.


8-Methoxy- and p-dimethoxy-1-aminoethylhetero-tetralins: synthesis and DA, 5-HT receptors affinities.

Two series of compounds, 8-methoxy- and p-dimethoxy derivatives of 1-aminoethylhetero-tetralins, I, were evaluated for D-2, 5-HT1 and 5-HT2 receptors affinity. No significant serotoninergic affinity was observed, whereas p-dimethoxy-derivatives 7b e 11b showed a moderate D-2 affinity.