Identification of a B2-bradykinin receptor linked to phospholipase C and inhibition of dopamine stimulated cyclic AMP accumulation in the human astrocytoma cell line D384. We have examined the activation of a phospholipase C signal transduction pathway by a B2-bradykinin receptor in the human astrocytoma cell line D384 and how this influences D1-dopamine receptor stimulated cyclic AMP accumulation. Addition of bradykinin to D384 cells resulted in a concentration-dependent (10(-11)-10(-6) M) increase in the accumulation of [3H]inositol phosphates and a similar concentration-dependent transient increase in specific [3H]beta-phorbol-12,13-dibutyrate binding which is indicative of translocation of protein kinase C from the cytosol to the membrane. Changes in intracellular Ca2+ of single cells, measured using the fluorescent indicator dye fura-2, indicated that bradykinin produced a rapid, but transient, increase in intracellular calcium. The Ca2+ response was largely independent of extracellular Ca2+ supporting the idea that receptor activation leads to mobilization of Ca2+ from intracellular stores. However, extracellular Ca2+ was required for a response to a rechallenge with bradykinin. The bradykinin B2-receptor agonist kallidin increased cytosolic Ca2+ in a similar manner to bradykinin. The Ca2+ response to bradykinin could be partially reduced in the presence of the B2-receptor antagonist [D-Arg0-Hyp,D-Phe7,beta-(2-Thienyl)-Ala5,8]-bradykinin, whereas the B1-receptor agonists (Des-Arg9]-bradykinin and [Des-Arg10]-kallidin were ineffective. Bradykinin was also found to attenuate dopamine stimulated cyclic AMP accumulation in D384 cells, at similar concentrations previously observed to stimulate the phospholipase C signal transduction pathway, in the presence of the phosphodiesterase inhibitor, rolipram. In contrast, no attenuation was observed in the presence of the phosphodiesterase inhibitor 1-isobutyl 3-methylxanthine, although the level of dopamine stimulated cyclic AMP observed was lower than in the presence of rolipram.(ABSTRACT TRUNCATED AT 250 WORDS)


Enantioselectivity of asocainol studied at different conditions: a novel approach to check the feasibility of molecular models of antiarrhythmic drug action. In terms of the "guarded receptor" hypothesis, changes in potency of Na+ channel blocking drugs reflect alterations in drug access to and/or egress from a compartment facing a binding site with constant affinity. Potency is therefore assumed to be determined by changes in drug diffusion, its mobility in the electric field, protonation etc. Hence, the potencies of enantiomers, i.e. compounds with identical physicochemical properties, should be influenced in a parallel manner by the condition. To test this prediction, actions of the enantiomers of the stereoselective antiarrhythmic drug asocainol were compared at various membrane potentials and stimulus frequencies. Several experimental models indicative of Na+ channel block were used: the elevation of the rectangular pulse stimulation threshold (RPT) and the suppression of alternating-current induced arrhythmia (ACT) were studied in guinea-pig atria. The reduction of the upstroke velocity of action potentials was measured in guinea-pig papillary muscles. The inhibition of whole-cell Na+ currents was investigated in isolated guinea-pig ventricular myocytes. In all these assays, (+)-asocainol was more potent than the (-)-enantiomer. Lowering the membrane potential and/or increasing the stimulus frequency enhanced the effects of both enantiomers. However, over a certain range of conditions, the potency of (+)-asocainol was more markedly affected than that of (-)-asocainol, indicating that the eudismic ratio between potencies of the two drugs is not constant. Accordingly, these findings are inconsistent with the guarded receptor hypothesis.


[Increased incidence of breast cancer in southeast Netherlands between 1960 and 1989]. Trends in incidence, stage distribution and mortality of breast cancer were determined in the Southeastern Netherlands in 1960-1989. First and second primary breast cancers were analyzed separately. The incidence and mortality rates were age-standardised according to the European Standard Population. The incidence rate of first primary invasive breast cancer increased from 50 per 100,000 women per year in 1960-61 to 96 in 1988-89, and of second primary invasive breast cancer from 3.2 in 1965-66 to 7.8 in 1988-89. The incidence rate of first primary ductal carcinoma in situ (DCIS) increased from 0.3 per 100,000 women per year in 1975-76 to 2.8 in 1988-89 and of second primary DCIS from 0.06 in 1975-79 to 0.4 per 100,000 in 1985-89. The increase in first invasive primaries was observed in all age groups, but mostly at age 75 and over. The percentage with stage I tumours of the patients with a first primary invasive breast cancer increased from 7% in the sixties to 27% in the eighties, whereas the percentage of stage III and IV tumours combined, decreased from 39% in the sixties to 26% in the eighties. While age-adjusted incidence of first primaries almost doubled in the past thirty years, breast cancer mortality rose only slightly. However, breast cancer mortality showed a proportionate increase of total mortality from 4.5% in 1970-71 to 6.3% in 1988-89.(ABSTRACT TRUNCATED AT 250 WORDS)


[Chest pain due to sumatriptan].

[Chest pain due to sumatriptan]. Since the registration of sumatriptan (Imigran) in May 1991, the Netherlands Centre for Monitoring of Adverse Reactions to Drugs received 13 reports of substernal chest tightness or pain attributed to its use. These concerned 11 women and two men with an average age of 41 years, who developed angina-like symptoms mostly within one hour after oral or subcutaneous administration. Electrocardiography (in three patients) and echocardiography (in one patient), performed after resolution of the symptoms, were normal. With the exception of one patient, none had experienced such symptoms before. Although sumatriptan is an effective anti-migrainous drug, cautious use is strongly advised. Sumatriptan is contraindicated in patients with coronary artery disease or variant angina pectoris.


[The course of a clinically treated depression in the elderly]. A study was performed of the course of major depressions (DSM-III) of elderly people who had been admitted to a psychogeriatric ward of a general psychiatric hospital. Patients were selected by searching the files. Patients with another diagnosis on axis I or a serious physical illness were excluded. The selected patients were traced and asked to participate in a follow-up investigation. From the files 38 patients were selected, of whom three had died in hospital. The remaining 35 patients included 28 women and seven men, with an average age of 78 years. After discharge seven of them died a natural death and six patients refused to participate. The remaining 22 patients were visited. From the files it appeared that 54% had completely recovered at discharge. Patients with a delusional depression had been hospitalised significantly longer, had been treated with more medicines and had less often been completely recovered at discharge. At the time of the follow up 64% had completely recovered but 32% had had a relapse. Patients with a delusional depression less often recovered completely. A remarkably low percentage of the population studied were able to live on their own. The results of this study are compared with foreign studies.


[ACTH-producing small-cell lung carcinoma]. Small-cell lung cancer may be accompanied by hypercortisolism due to secretion of adrenal stimulating peptides. This occurs almost exclusively in case of extensive disease. The neuroendocrine cell, from which these tumours may originate, plays an important part in the production of these peptides. Four patients are described with small-cell lung cancer and clinical presentation of hypercortisolism, particularly manifesting itself with hypokalaemia.


[Human papillomavirus and intraepithelial cervical cancer after renal transplantation. Apropos of 2 observations]. Therapeutic immunosuppression applies to an increasing number of patients, exceeding largely the only transplant recipients. Among its detrimental carcinological complications are the uterine cervix intraepithelial neoplasia whose strong relationship with the lesions due to human papillomavirus is well known. They need an early and regular follow-up of the concerned women, by cervical smear tests, colposcopic assessment and histological examinations, to undertake in time the adequate treatment, without forgetting to give some preventive advice.


Muscimol and flunitrazepam binding sites in a subcellular fraction enriched in rat cerebellar glomeruli. In the internal granular layer of the cerebellar cortex the polysynaptic complexes called glomeruli consist mainly of homogeneous populations of glutamatergic and GABAergic synapses, both located on granule cell dendrites. A subcellular fraction enriched in glomeruli was prepared from rat cerebellum, and the distribution of GABAA and of benzodiazepine binding sites between membranes derived from this fraction (fraction G) and from a total cerebellar homogenate (fraction T) was studied. The benzodiazepine and GABA binding sites were measured by the binding of agonists [3H]flunitrazepam and [3H]muscimol, respectively. The results indicate that both binding sites are present, but only slightly enriched, in the glomerular synapses. We found a muscimol/flunitrazepam binding site ratio of two, which is consistent with the enrichment of muscimol binding sites in the granular layer shown by both autoradiographic with radioactive glutamatergic ligands and in situ hybridization experiments respectively.


[3H]muscimol and [3H]flunitrazepam binding sites in the developing cerebellum of mice treated with methylazoxymethanol at different postnatal ages. Two models of perturbed cerebellar ontogenesis were obtained by a single administration of methylazoxymethanol (MAM), a potent antimitotic agent, to mouse pups either on the day of birth (MAM0 mice) or at postnatal day 5 (MAM5 mice). The alterations of the cerebellar GABAergic system were studied by measuring glutamic acid decarboxylase activity, [3H]muscimol binding sites, which are known to be concentrated in the GABAA receptors in the internal granular layer, and [3H]flunitrazepam binding sites, which are more abundant in the molecular layer. The primary target of the antimitotic agent are the precursors of the glutamatergic and GABAceptive granule cells. In both models GABAergic structures, as revealed by GAD activity measurements, appear to be relatively spared, and recovery of granule cell numbers occurs during development in MAM5 mice. In MAM treated mice the number of [3H]muscimol binding sites (on a per cerebellum basis) decrease as the number of granule cells decrease, although some recovery occurred in MAM5 mice, but not in MAM0 mice. In MAM5 mice, [3H]flunitrazepam binding sites (on a per cerebellum basis) were relatively unaffected, while they were decreased significantly, but to a lesser extent than [3H]muscimol binding sites, in MAM0 animals. The more significant reduction of granule cell numbers and the cytoarchitectural disruption resultant from the more precocious application of the antimitotic appear responsible for the significant alteration and lack of recovery in MAM0 mice.


Increase in nucleoside diphosphatase in rat brain striatum lesioned with kainic acid. The activity of ammoniagenesis from guanine nucleotides was found to increase significantly in rat brain after infusion of kainic acid into the striatum. Among the enzymes involved in degrading guanine nucleotides, nucleoside diphosphatase was markedly increased in the lesioned striatum. The enzyme activity began to increase 2 days after the infusion, and reached the maximum on the 13th day, the level being 4 times as high as that of the intact contralateral region. The increased activity was due to Type L enzyme, judging from its substrate specificity. Puromycin and cycloheximide inhibited this increase, indicating that the increased activity resulted from an increase in the net synthesis of the enzyme. These findings suggest that Type L NDPase might play some important roles in gliosis after neuronal lesion.


N-type calcium channels are involved in the dopamine releasing effect of nicotine. Mouse striatum was incubated with [3H]dopamine ([3H]DA) and superfused with and the tritium efflux induced by nicotine, electrical stimulation, or simultaneous nicotine and electrical stimulation was measured, to characterize the role of different Ca2+ channels in the transmitter release. Nicotine stimulation and electrical stimulation exerted additive effects on tritium efflux. Separation of the released radioactivity on alumina columns indicated that nicotine or electrical stimulation increases the release of [3H]DA and that the outflow of 3H-labeled metabolites was similar with the two different stimulation procedures. Removal of Ca2+ from the superfusate resulted in a marked reduction in the tritium release evoked by nicotine, whereas the electrical stimulation-evoked tritium release was completely dependent on external Ca2+. The L- and N-type calcium channel blockers omega-conotoxin GVIA and Cd2+ inhibited the tritium release from the striatum evoked by either nicotine or electrical stimulation, whereas the L-type and T-type channel blockers diltiazem and Ni2+ did not alter release of [3H]DA. We conclude that N-type voltage-sensitive Ca2+ channels participate in striatal dopamine release, and we speculate that nicotinic receptor-operated ion channels permeable to cations such as Ca2+ and N-type voltage-sensitive calcium channels may simultaneously open up, and they additively increase free intracellular Ca2+ concentration.


Differential time course activation of the brain stem catecholaminergic groups following chronic adrenalectomy. The activity of the brain stem catecholaminergic (CA) cell groups of the ventrolateral (A1C1) and dorsomedial (A2C2) medulla that are known to contain primarily nonadrenergic neurones (A1 and A2) and a smaller proportion of adrenergic cells (C1 and C2) as well as the noradrenergic group locus ceruleus (LC) in the dorsal pons was determined at various times up to 16 days following surgical adrenalectomy. The activity of the CA cell groups was estimated by the rate of tyrosine hydroxylation in vivo that was assessed by measuring the 3.4-dihydroxyphenylalanine (DOPA) accumulated 20 min following administration of DOPA decarboxylase inhibitor NSD 1015. In the medullary nuclei noradrenaline content was found around 40- up to 70-fold the adrenaline content. This result was taken as evidence that the noradrenergic cells are likely to provide the main contribution to the tyrosine hydroxylation rate that we measured. Endogenous DOPA content represented between 2 and 10% of the noradrenaline content. NSD 1015 induced an accumulation of DOPA that was linear for at least 20 min and reached at this time more than 10-fold the endogenous level. While no modification of the in vivo tyrosine hydroxylation rate was observed in the LC, a significant increase was found in both medullary groups following adrenalectomy. In the A1C1 group it was detected 8 days after surgery and was then maintained with a maximum that represented up to a 60% increase over the basal value. In the A2C2 group the activation was slightly delayed and less marked. Increase in ACTH level occurred much earlier: it was about 70% of the maximal level already 4 days following adrenalectomy.(ABSTRACT TRUNCATED AT 250 WORDS)


Modulation of pulsatile LH secretion by baclofen, a selective GABAB receptor agonist, in ovariectomized rats. The effect of activation of central gamma-aminobutyric acidB (GABAB) receptors on pulsatile luteinizing hormone (LH) secretion was examined in unanesthetized ovariectomized rats. Intraventricular injections of 2 or 10 micrograms baclofen, a selective GABAB receptor agonist, usually arrested the existing pulsatile LH secretion for a short period, then LH pulses resumed with reduced frequencies and increased amplitudes. The increase in the pulse amplitude was not necessarily accompanied by a reduction in the pulse frequency, but was observed even after nonelongated interpulse intervals in animals receiving the GABA agonist. Mean LH levels showed complex bi- and triphasic fluctuations after the central administration of high and low doses of baclofen, respectively. It was concluded that central GABAB receptors are involved in the modulatory mechanism affecting the LH pulse frequency and amplitude in the ovariectomized rat.


Participation of alpha 1-adrenergic receptors in the secretion of hypothalamic corticotropin-releasing hormone during stress. The role of alpha 1-adrenergic receptors in the secretion of corticotropin-releasing hormone (CRH) during stress was studied by immunohistochemical analysis of the CRH content of the median eminence (ME) after intracerebroventricular (icv) administration of the alpha 1-adrenergic agonist, methoxamine, or the antagonist, prazosin, in rats pretreated with colchicine. Immunohistochemical staining was performed by the peroxidase technique on 40 microns free-floating sections using a polyclonal antibody specific for CRH. In the first experimental model, rats were implanted with icv cannulae and adapted to the experimental conditions by daily handling and icv injection of artificial CSF. Colchicine (75 micrograms) was administered through the cannulae 6 h before the experiment, conditions in which axonal transport was blocked with little change in basal immunostaining. Two hours after immobilization stress or a single injection of methoxamine (100 micrograms, icv), there was a marked decrease in CRH immunoreactivity throughout the ME, reflecting release of the neuropeptide into the portal circulation. The decrease in CRH immunostaining following immobilization was largely prevented by icv injection of the alpha 1-adrenergic antagonist, prazosin. In the second experimental model, rats were sacrificed 48 h after icv colchicine injection, conditions in which colchicine acts as a stressor and causes marked depletion of irCRH from the ME. This chronic effect of colchicine was also partially prevented by administration of prazosin, 400-ng injection 5 min prior to colchicine, followed by a continuous icv mini-pump infusion of prazosin, indicating that alpha 1-adrenergic stimulation contributes to the action of colchicine.(ABSTRACT TRUNCATED AT 250 WORDS)


Arrested maturation of cerebral neurons, axons and myelin: a new familial syndrome of newborns. A new lethal familial syndrome of unknown etiology is described in two male siblings who died in the newborn period. Both had corneal edema and were hypotonic, requiring assisted ventilation at birth. Neuropathological findings included an immature appearance of neocortical neurons, with cortical architecture similar to that normally seen in an infant of 5 months gestational age. Axons and myelin were absent in the cerebral and cerebellar white matter, and also in descending white matter tracts of brainstem and spinal cord. Subacute inflammation was seen in the anterior horns of the spinal cord in both cases, although there was no evidence of inflammation elsewhere in the nervous system. Electron microscopy of endothelial cells from brain, spinal cord and a number of other tissues of the second sibling showed tubuloreticular inclusions (TRIs). There are no known previous reports of similar neuropathology. Future recognition of this condition will be important for genetic counselling.


Dissociation of nitric oxide generation and kainate-mediated neuronal degeneration in primary cultures of rat cerebellar granule cells. In the presence of physiological concentrations of Mg2+ and in glycine-free buffer, the relationship between KA-mediated generation of NO and neurotoxicity in cultures of cerebellar granule cells of the rat was examined. The neuronal damage elicited by KA was not dependent on the presence of L-arginine, a precursor of NO, since neither the potency nor magnitude of KA-mediated cell death was altered in either the absence or presence of exogenously applied L-arginine. Similarly, with the exception of 4-hydroxy-azobenzene-4'-sulfonic acid, disodium salt dihydrate (HBS), the salt associated with NG-monomethyl-L-arginine (di-(p-hydroxyazobenzene-p'-sulfonate) (MA(HBS)), treatment with several different competitive NO synthetase inhibitors did not provide protection against the toxicity of KA. However, the ability of KA to induce neuronal damage was significantly decreased in cerebellar granule cells treated with either HBS or alpha-tocopherol (VE). On the basis of these results, it is concluded that the generation of free radicals may be involved in the process of KA-elicited neuronal death in cultures of cerebellar granule cells but that this is unrelated to the synthesis of NO. This conclusion agrees with both in vivo and in vitro studies, implicating the involvement of free radicals in non-NMDA mediated neuronal damage.


Effects of d-amphetamine on dopaminergic neurotransmission; a comparison between the substantia nigra and the striatum. The effects of d-amphetamine (d-AMP) on dopaminergic neurotransmission in the cell body/dendritic region of the nigrostriatal pathway, the substantia nigra, have been investigated and compared to the effects obtained in the terminal region of the pathway, the striatum. The rate of synthesis of dopamine (DA) was quantified as accumulation of 3,4-dihydroxyphenylalanine (DOPA), after inhibition of aromatic L-amino acid decarboxylase with 3-hydroxybenzyl-hydrazine (NSD 1015). As measures of the metabolism of DA the concentrations of the metabolites of DA, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 3-methoxytyramine (3-MT) were determined. As indices of release of DA, the accumulation of 3-methoxytyramine (3-MT), after inhibition of monoamine oxidase with pargyline and the disappearance of DA, after inhibition of its synthesis with alpha-methyl-p-tyrosine were assessed. d-Amphetamine insignificantly increased the concentration of DA in the striatum but profoundly decreased it in the substantia nigra (to 60% of controls). Both in the striatum and in the substantia nigra treatment with d-AMP induced clearcut decreases of the concentrations of DOPAC. Also the concentration of HVA was profoundly decreased in the striatum but only marginal effects on HVA were observed in the substantia nigra. In both structures of the brain, d-AMP increased the concentration of 3-MT. Depending on the dose, d-AMP increased or had no effect on the accumulation of DOPA in the striatum but consistently decreased it in the substantia nigra. In the striatum, d-AMP increased the pargyline-induced accumulation of 3-MT, without affecting the concentration of DA. However, in the substantia nigra the concentration of DA was profoundly decreased (to 50% of controls) in combination with unaltered accumulation of 3-MT, unless the rats were pretreated with haloperidol. If so, the effects of d-AMP on the concentration of DA and accumulation of 3-MT were the same in the substantia nigra and in the striatum. The results indicate that d-AMP depleted stores of DA in the substantia nigra, due to its releasing action, in combination with its decreasing effect on the rate of synthesis of DA. The decrease in rate of synthesis of DA is suggested to be due to the d-AMP-induced decrease in the firing rate of dopaminergic neurones.


D1 and D2 dopamine receptors stimulate hypothalamo-pituitary-adrenal activity in rats. A stimulatory role for endogenous dopamine (DA) in the regulation of hypothalamo-pituitary-adrenal activity has previously been demonstrated. In the present study, the roles of D1 and D2 subtypes of DA receptors in the regulation of activity of the hypothalamo-pituitary-adrenal axis were investigated. The intraperitoneal administration of either the D1 agonist, SKF 383393 (1-phenyl-2,3,4,5 tetrahydro-(iH)-benzazepine-7,8diol HCl, 5-20 mg/kg) or the D2 agonist quinpirole (0.05-1 mg/kg) dose-dependently elevated both adrenocorticotropic hormone (ACTH) and corticosterone (CS) in serum. Similarly, administration of either SKF 38393 or quinpirole (1-100 micrograms) into the third ventricle dose-dependently elevated ACTH in serum. The response of ACTH to intraperitoneal SKF 38393 was blocked by pretreatment with the D1 antagonist SCH 23390 (1-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5 tetrahydro-1H-3-benzazepine, 0.25 mg/kg, i.p.) but not by the D2 antagonist sulpiride (50 mg/kg, i.p.). The response of ACTH to intraperitoneal injection of quinpirole was blocked by pretreatment with sulpiride and attenuated slightly by pretreatment with SCH 23390. Further, the co-administration of sub-maximum doses of SKF 38393 and quinpirole caused additive increases in ACTH in serum. These results suggest that both D1 and D2 subtypes of DA receptors contribute to the dopaminergic regulation of function of the hypothalamo-pituitary-adrenal axis and support a role for DA neurons in the hypothalamus in this response. Further, these findings suggest that the D1 and D2 receptors, mediating the response of the hypothalamopituitary-adrenal axis are not tightly coupled.(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of receptor blockers on brain natriuretic peptide-32-induced action on passive avoidance behavior in rats. The effects of several doses of porcine brain natriuretic peptide-32 (pBNP-32) administered into the lateral brain ventricle were tested as regards the consolidation of passive avoidance learning in rats. The peptide was found to increase the passive avoidance latency in a dose-dependent manner. In order to clarify which transmitter systems might be involved in the action of pBNP-32, the experimental animals were pretreated with different receptor blockers in selected doses which did not influence the behavioral paradigm. Four of the receptor blockers (haloperidol, atropine, phenoxybenzamine and propranolol) effectively blocked the action of the peptide on the consolidation of passive avoidance learning. The other three (naloxone, bicuculline and methysergide) were ineffective. The results suggest that dopaminergic, cholinergic and alpha- and beta-adrenergic mediations might be involved in the effects of pBNP-32 on the consolidation of passive avoidance learning in rats.


Intravenous administration of tumor necrosis factor-alpha stimulates corticotropin releasing hormone secretion in the push-pull cannulated median eminence of freely moving rats. Utilizing push-pull perfusion, we examined the effects of intravenous (iv) administration of human recombinant tumor necrosis factor (TNF)-alpha on the levels of plasma adrenocorticotropin (ACTH) and corticotropin releasing hormone (CRH) in the median eminence (ME) of freely moving male rats. The ME was perfused with artificial cerebrospinal fluid between 11:00 and 14:00 h, and perfusates and blood samples were collected every 20 min. TNF-alpha (1.0 microgram), but not vehicle only, given as an iv bolus at 12:00 h significantly stimulated both plasma ACTH and ME-CRH. The increase in ME-CRH clearly preceded that of plasma ACTH. This is the first to characterize the temporal profile of CRH secretion in the ME after iv administration of TNF-alpha to freely moving rats. These in vivo data strongly suggest that TNF-alpha stimulates ACTH secretion, at least in part, by triggering hypothalamic CRH release. In addition, combined with our previous data obtained by iv administration of human recombinant interleukin-1 under the same experimental condition, the present study also suggests that iv injected TNF-alpha and interleukin-1 may share a common site of action in the brain, such as the ME, to stimulate CRH secretion.


Localization of vasoactive intestinal polypeptide-receptor-immunoreactivity in human salivary glands. A monoclonal VIP-receptor antibody derived from a human adenocarcinoma cell line (HT 29) was used in combination with immunogold silver staining for the immunohistochemical demonstration of VIP-receptor (rec) immunoreactivity (IR) in paraffin-embedded human salivary glands (parotid, palatal, labial glands). VIP-rec-IR was localized to mucous endpieces of labial and--to a lesser extent--palatal glands, intercalated ducts of the parotid gland, and excretory ducts of all glands investigated. The findings correlate well with known effects of VIP on mucous release and electrolyte transport in salivary glands. Lack of VIP-rec-IR at serious acini may point to immunologically different receptor subtypes in these glands.


Affinity of R 396, an NK-2 tachykinin receptor antagonist, for NK-2 receptors in preparations from different species. We have assessed the affinity of R 396 (Ac. Leu-Asp-Gln-Trp-Phe-Gly NH2) in a number of NK-2 tachykinin receptor bearing-tissues from several species. The cyclic analog of R 396, (MEN 10354) was less potent and selective than the linear hexapeptide at NK-2 tachykinin receptors subtypes in the rabbit pulmonary artery and hamster trachea. The affinity of R 396, as measured by a smooth muscle contraction assay and a radioligand binding assay, was higher (about 10 fold) for NK-2 receptors expressed in hamster tissues (urinary bladder, stomach and trachea) than in rat tissues (urinary bladder, vas deferens, colon and stomach) and a further drop in affinity was observed in bovine tissues (urinary bladder and stomach) or rabbit bronchus. The results are discussed in relation to the proposed existence of NK-2 receptor subtypes and raise the question of the existence of species-related differences as compared to the existence of true receptor subtypes.


Interaction of amyloid beta protein (25-35) with tachykinin receptors. Amyloid beta protein (25-35) failed to significantly interact with tachykinin NK1 (rat forebrain, guinea-pig ileum) NK2 (rabbit pulmonary artery, hamster trachea) or NK-3 (guinea-pig cortex) receptors, as determined by radioligand binding and functional assays. A weak interaction (Ki 14.8 microM) was detected with NK2 receptors in rat small intestine. It appears unlikely that direct interaction with tachykinin receptors may account for the reported ability of amyloid beta protein (25-35) to affect neuronal survival.


Structural requirements for the effects of neuropeptide Y on the hypothalamic-pituitary-adrenal axis in the dog. Neuropeptide Y (NPY), administered into the third cerebral ventricle of the dog stimulates plasma ACTH and cortisol secretion. To further investigate the structure-activity relationships of this action, we examined the effect of COOH-terminal fragment, NPY 19-36 and its analogues, NPY-(1-36)-OH (deamidated NPY) and avian pancreatic polypeptide (APP) on ACTH-cortisol secretion following intracerebroventricular (i.c.v.) injection in the dog. NPY (1.19 nmol) evoked a significant increase in the secretion of both plasma ACTH and cortisol. However, NPY 19-36 and NPY-(1-36)-OH each failed to increase plasma ACTH and cortisol secretion at doses of 1.19-11.9 nmol injected i.c.v. APP was less active than NPY. These data demonstrate that the entire NPY molecule is required for the full expression of the stimulatory effect of NPY on the secretion of ACTH and cortisol.


Intracerebral malignant fibrous histiocytoma: case report and review of the literature. A 50-year-old woman developed an intracerebral malignant fibrous histiocytoma at the site of a previously clipped aneurysm in the right temporal lobe. This tumor rarely originates within the brain. Review of the literature suggests that postoperative and/or postirradiation effects may contribute to the growth of this particular tumor.


Autoradiographic distribution of receptors to FLFQPQRFamide, a morphine-modulating peptide, in rat central nervous system. The neuropeptide FLFQPQRFamide is a structure related to FMRFamide which is able to inhibit the effects of both endogenous and exogenous opiates. This morphine-modulating activity is mediated via the stimulation of specific FLFQPQRFamide receptors, different from opiate receptors. In vitro quantitative receptor autoradiography was performed on frozen sections of rat central nervous system to characterize binding properties and visualize FLFQPQRFamide receptors using the specific ligand [125I]YLFQPQRFamide, a radio-iodinated analogue of FLFQPQRFamide. [125I]YLFQPQRFamide appeared to interact reversibly with a single class of binding sites (KD = 0.2 nM). The specific binding represented 80% of the total binding at 0.05 nM, the FLFQPQRFamide concentration used in this mapping study. Sites labelled with [125I]YLFQPQRFamide were distributed heterogeneously within the brain and spinal cord. A high density of FLFQPQRFamide binding sites was detected in the most external layers of the dorsal horn of spinal cord and various nuclei of pons and medulla including trigeminal, dorsal tegmental and reticular nuclei. Nucleus of solitary tract, parabrachial, ambiguous and facial nuclei are also intensively labelled. Some structures of mesencephalon and diencephalon exhibited a high density of FLFQPQRFamide binding sites: central gray, raphe nuclei and thalamic nuclei such as parafascicular, laterodorsal, central median, paratenial and paraventricular nuclei. Suprachiasmatic and mammillary nuclei, lateral, posterior and anterior areas of hypothalamus and medial preoptic area exhibited high labelling. FLFQPQRFamide binding sites were also seen in some structures of the dopaminergic meso-cortico-limbic system including ventral tegmental area, cingulate cortex, lateral septum and the head of the caudate-putamen. Dense labelling appeared in the presubiculum of hippocampus. The dissimilar mapping of FLFQPQRFamide and opiate brain receptors confirms our previous pharmacological findings in FLFQPQRFamide binding studies on rat spinal cord membranes, showing that FLFQPQRFamide receptors are different from opiate receptors. There was a good correspondence between localization of binding sites and that of the putative endogenous peptide. Both occur in brain areas previously associated with analgesic action of opiates. However, the mapping of FLFQPQRFamide receptors in the central nervous system suggests that the FLFQPQRFamide system could be implicated in other physiological functions.


Delta- and kappa-opioid agonists inhibit plasma extravasation induced by bradykinin in the knee joint of the rat. We used an experimental model of neurogenic inflammation, plasma extravasation induced by bradykinin or capsaicin, to study the effect of receptor-selective opioid agonists on plasma extravasation. Plasma extravasation was induced in the knee joint of the rat by continuous perfusion of either bradykinin (160 ng/ml), an inflammatory mediator produced at sites of tissue injury, that produces plasma extravasation significantly dependent on the sympathetic postganglionic neuron, or capsaicin (5 mg/ml), a C-fiber excitotoxin, that induces plasma extravasation that is dependent on both primary afferents and sympathetic post-ganglionic neurons. When selective delta-((d-Pen2,5)-enkephalin) or kappa-(trans-3,4-dichloro-N-methyl-N[2-(- pyrolidinyl)cyclohexyl]benzeneacetamide; U50,488H) opioid agonists were perfused with bradykinin, plasma extravasation was significantly attenuated. Co-perfusion of the non-selective opioid antagonist naloxone (1 microM), reversed this opioid-induced inhibition of bradykinin-induced plasma extravasation. In contrast, co-perfusion of a selective mu-opioid agonist (Tyr-d-Ala-Gly-NMe-Phe-Gly-ol) did not reduce bradykinin-induced plasma extravasation. Tyr-d-Ala-Gly-NMe-Phe-Gly-ol was, however, able to completely inhibit the plasma extravasation produced by capsaicin. These results suggest that delta- and kappa-, but not mu-selective opioids inhibit bradykinin-stimulated plasma extravasation, while a mu-selective opioid inhibits primary afferent-dependent plasma extravasation. Therefore, inhibition of neurogenic plasma extravasation by receptor-selective opioids may depend on the relative contribution to plasma extravasation of unmyelinated afferent and sympathetic postganglionic neuron terminals. Our findings can also explain, in part, the variation in anti-inflammatory effects of receptor-selective opioids reported in different inflammatory conditions.


Differential effects of peripheral damage on vibrissa-related patterns in trigeminal nucleus principalis, subnucleus interpolaris, and subnucleus caudalis. Histochemistry for cytochrome oxidase reveals a vibrissa-related pattern in trigeminal nucleus principalis, subnucleus interpolaris, and the magnocellular portion of subnucleus caudalis. This pattern is apparent in late fetal animals and is disrupted by transection of the infraorbital nerve on the day of birth. We recently reported results suggesting that the cytochrome oxidase pattern reflects primary afferent-induced clustering of second order neurons in all of these nuclei. If this conclusion is correct, it should follow that primary afferent lesions made after the cytochrome oxidase pattern became established in the brainstem might have little effect upon it. Accordingly, we transected the infraorbital nerve (the trigeminal branch that supplies the vibrissae) on postnatal days 0-10 and evaluated the vibrissa-related pattern in the brainstem with cytochrome oxidase histochemistry at varying intervals after these lesions. If the infraorbital nerve was sectioned on postnatal days 0-2, the vibrissa-related pattern was absent in trigeminal nucleus principalis, and both subnucleus interpolaris and caudalis. If such lesions were made after postnatal day 9, there was no appreciable effect upon the cytochrome oxidase pattern in any portion of the trigeminal brainstem complex. However, if lesions were made between postnatal days 3 and 8, the density and clarity of the cytochrome oxidase staining pattern were reduced in interpolaris and caudalis, but not in principalis. This difference was not due to differential transganglionic degeneration in these nuclei. Tracing with horseradish peroxidase demonstrated qualitatively equivalent primary afferent losses in principalis, interpolaris, and caudalis. Immunocytochemistry with a monoclonal antibody directed against parvalbumin also demonstrated a vibrissa-related pattern of cell bodies in principalis and interpolaris in rats killed on postnatal day 9 or later ages. The combination of retrograde tracing and immunocytochemistry revealed that the parvalbumin-immunoreactive neurons in principalis projected to thalamus while those in interpolaris were not labelled by tracer injections into the thalamus, midbrain, cerebellum or spinal cord. Infraorbital nerve transections made as late as postnatal day 8 resulted in a sharp decrease in the staining of parvalbumin-positive neurons in interpolaris, but not in principalis. Lesions made on postnatal day 10 had no qualitative effect upon parvalbumin-positive neurons in any portion of the trigeminal brainstem complex. The results of this study support the conclusion that the vibrissa-related cytochrome oxidase pattern in principalis becomes independent of primary afferent input at a very short interval after its initial appearance. In contrast, the patterns in more caudal portions of the trigeminal brainstem complex require maintenance of primary afferent input for a much longer postnatal period.(ABSTRACT TRUNCATED AT 400 WORDS)


Characterization of L-glutamate and kainate binding sites in the brain of a freshwater fish, Telapilia monsanbica. [3H]Kainate and L-[3H]glutamate binding sites in a rich source of kainate binding sites, fish brain, have been thoroughly analysed here for the purpose of studying the correlation between kainate binding sites and L-glutamate receptors in vertebrate CNS. The brain of a freshwater fish, Telapilia monsanbica, was found to contain three types of kainate binding sites: Type 1 sites (Kd = 1050 +/- 380 microM, Bmax = 4 +/- 4 pmol/mg), Type 2 sites (Kd = 133 +/- 20 nM, Bmax = 190 +/- 20 pmol/mg), and Type 3 sites (Kd = 23 +/- 15 nM, Bmax = 28 +/- 19 pmol/mg). The dissociation constants of L-glutamate to Type 1, 2 and 3 sites were, respectively, 0.28 +/- 0.04, 5.5 +/- 0.2 and 137 +/- 28 microM. Pharmacological characterization of these binding sites showed that Type 1 and 2 sites, respectively, corresponded to N-methyl-D-aspartate-subtype L-glutamate receptors and non-N-methyl-D-aspartate L-glutamate receptors. Autoradiographic studies showed that Type 1 and 2 sites were distributed widely in fish brain, indicating the involvement of L-glutamate receptors in various brain functions. Type 3 sites, on the other hand, were relatively insensitive to most endogenous amino acids and were only found in the molecular layer of cerebellum and torus longitudinalis. Type 3 sites possibly representing a distinctive class of receptor has been suggested by the results.


Influence of small cardioactive peptide (b) on the efficiency of synaptic transmission and the excitability of command neurons of the defensive behavior of the edible snail. The application of a small cardioactive peptide (SCPb) (mCPb) in a concentration of 5 x 10(-8) mole/l to a physiological solution washing the isolated central nervous system of the edible snail leads to an increase in the amplitude of the summated EPSP evoked by rhythmic stimulation of the intestinal nerve in the command neurons of defensive behavior by 55 +/- 20%. In addition to the change in the magnitude of the synaptic input, SCPb induces an increase in the excitability of the same neurons by 92 +/- 57%, tested intracellularly. These influences may lead to a total decrease in the threshold of the reaction of the neuronal network of the defensive behavior in response to stimulation. The possibility of the integration of integral behavior with the aid of peptides which are effective in very small concentrations is discussed.


Localization of AMPA receptors in the hippocampus and cerebellum of the rat using an anti-receptor monoclonal antibody. The primary amino acid sequences of the kainate binding proteins from the amphibian and avian central nervous systems are homologous with the functional alpha-amino-3-hydroxyl-5-methyl-isoxazole-4-propionate receptors that have been cloned from rat brain. In this study, we have analysed the anatomical and subcellular distribution of the alpha-amino-3-hydroxyl-5-methyl-isoxazole-4-propionate receptors in the rat hippocampus and cerebellum, using a monoclonal antibody that was raised against a kainate binding protein purified from frog brain. Immunoblots of rat hippocampus and cerebellum, and membranes from COS cells transfected with rat brain alpha-amino-3-hydroxyl-5-methyl-isoxazole-4-propionate receptor cDNAs (GluR1, GluR2, or GluR3) showed a major immunoreactive band migrating at a relative molecular weight of 107,000. In the cerebellum, an additional immunoreactive protein of approximately 128,000 mol. wt was also seen on immunoblots probed with the antibody. The distribution of this protein is apparently restricted to the cerebellum since the 128,000 mol. wt band was not present in other brain areas examined. The identity of the 128,000 mol. wt cerebellar protein is not known. Immunocytochemical analyses of the hippocampus demonstrated that alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate receptor subunits are present in the cell bodies and dendrites of pyramidal cells. The granule cells were also immunostained. All of the pyramidal cell subfields were heavily labeled. In the pyramidal cell bodies, a high level of immunoreactivity was observed throughout the cytoplasm. In the cerebellum, the Purkinje cell bodies and dendrites also displayed very high levels of immunoreactivity. In addition to the Purkinje neurons, the Bergmann glia and some Golgi neurons were clearly immunostained. Subcellular fractionation and lesioning experiments using the excitotoxin domoic acid indicated that the alpha-amino-3-hydroxyl-5-methyl-isoxazole-4-propionate receptor subunits were associated with postsynaptic membranes. Direct visualization of the immunoreactivity using electron microscopy confirmed the postsynaptic localization of the staining in the dendritic areas in both the hippocampus and the cerebellum. Thus, unlike the kainate binding proteins, which are found primarily extrasynaptically in the frog and on glial cells in the chicken cerebellum, the GluR1, GluR2, and GluR3 receptor subunits are localized to the postsynaptic membrane in the dendrites of neurons in the rat central nervous system.


Hippocampal excitatory amino acid receptors in elderly, normal individuals and those with Alzheimer's disease: non-N-methyl-D-aspartate receptors. Quantitative receptor autoradiography was used to examine the density and distribution of [3H]kainic acid and [3H]alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) binding sites in the hippocampal formation and parahippocampal gyrus obtained at autopsy from 10 Alzheimer's disease and eight normal control individuals. In control and Alzheimer's disease individuals, [3H]kainic acid saturation binding analysis in the outer molecular layer of the dentate gyrus fitted a single-site model. Added calcium ions did not alter the density of [3H]kainic acid binding in the human tissues. These results suggest that calcium-sensitive high-affinity kainic acid binding sites are not present in the human brain in contrast to kainic acid receptors in the rat brain. [3H]AMPA binding was also slightly different in the human brain as compared to the rat, being greatest in the inner third as compared to the outer two-thirds of the dentate gyrus molecular layer. In both control and Alzheimer's disease individuals, [3H]kainic acid and [3H]AMPA binding densities were similar at anterior and posterior levels of the hippocampal formation. In Alzheimer's disease patients, there was a significant increase in [3H]AMPA binding in the infragranular layer. In some, but not all Alzheimer's disease patients, there was an increase in [3H]kainic acid binding densities in the outer half of the dentate gyrus molecular layer. The same individuals which exhibited an increase in [3H]kainic acid binding in the outer molecular layer also displayed increased [3H]AMPA binding in the hilar region. Similar alterations in [3H]kainic acid binding have been observed in rats which had received fimbria-fornix lesions, a model of chronic epilepsy and in individuals with temporal lobe epilepsy. Advanced Alzheimer's disease patients are at risk of developing seizures. The results suggest that several factors including cortical and subcortical pathology and seizure activity may contribute to the alterations in [3H]kainic acid and [3H]AMPA binding observed in the hippocampal formation in Alzheimer's disease.


Pseudo-Bartter's syndrome from surreptitious diuretic intake: differential diagnosis with true Bartter's syndrome. Five patients with pseudo-Bartter's syndrome from surreptitious diuretic abuse were compared with six patients with true Bartter's syndrome, diagnosed as a normotensive, hyperreninaemic, hypokalaemic metabolic alkalosis with normal urine chloride excretion, low CH2O/(CH2O+CCl) ratio during maximal water diuresis and negative urine screen for diuretics. The latter was positive for frusemide in four and for hydrochlorothiazide in the remaining pseudo-Bartter's patients. The two groups of patients did not differ as for plasma Na+, Cl-, K+, HCO3-, renin, and aldosterone, while uric acid and Mg2+ were greater in pseudo-Bartter's patients. Daily and fasting urine Na+, Cl- and K+ excretion were less in pseudo-Bartter's patients; however, there was substantial overlap of values between the two groups. Fractional distal solute reabsorption during maximal water diuresis was low in the six patients with Bartter's syndrome and in two pseudo-Bartter's patients; thus, this parameter could not be taken as a specific diagnostic marker of Bartter's syndrome. Frusemide administration, 40 mg i.v., induced a brisk increase of urine flow (11.7-21.8 ml/min), UOsm (148-186 mOsm/kg H2O) and FENa (14.6-24%) in Bartter's syndrome, but not pseudo-Bartter's patients; in all pseudo-Bartter's patients frusemide-induced changes of UOsm (13-97) and FENa (-0.5 to 10.2) were markedly less than in Bartter's syndrome patients. Frusemide resistance in pseudo-Bartter's patients was most probably related to diuretic-induced ECF volume contraction and increased proximal tubule solute reabsorption; in fact fractional lithium clearance (FELi, a marker of post-proximal solute delivery) was low in pseudo-Bartter's, but not in Bartter's syndrome patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantification of fetal antigen 2 (FA2) in supernatants of cultured osteoblasts, normal human serum, and serum from patients with chronic renal failure. Fetal antigen 2 (FA2) was found within the cytoplasm of osteoblasts and in osteoid material, in a bone biopsy with morphological changes of renal osteodystrophy. An ELISA technique for FA2 quantification was developed, and the specificity confirmed by comparison with electroimmunoassay. The intra- and interassay coefficient of variations (%) were 8.3 and 9.7 respectively, and the detection limit 0.0004 arbitrary units FA2/1 using second-trimester human amniotic fluid as reference (1 AU FA2/1). FA2 was detected in serum-free supernatants from osteoblast cultures. Following size chromatography, the FA2 distribution (two peaks eluted corresponding to Mw 30 kDa and 100 kDa) in serum from a patient with chronic renal failure complicated with secondary hyperparathyroidism and in human amniotic fluid were identical. Probably due to the detection limit, only one peak fraction (30 kDa) was seen in normal human serum. Significantly greater FA2 concentrations were found in sera (n = 14) from patients with chronic renal failure (median: 11.9 mAU FA2/1; range: 5.2-49.0 mAU FA2/1) compared to normal healthy individuals (n = 23) (median: 4.1 mAU FA2/1; range 2.4-9.4 mAU FA2/1) (P less than 0.00001). A close correlation was found between serum FA2 and alkaline phosphatase (R(s) = 0.761; P = 0.006), c-terminal fraction of PTH (R(s) = 0.872; P = 0.003) and intact PTH(1-84) (R(s) = 0.904; P = 0.011) in the haemodialysis patients. These data indicate that FA2 is synthesized by osteoblasts and may represent a new marker for metabolic bone changes.


Outcome of patients with secondary amyloidosis in dialysis treatment. Between the years 1974 and 1987, 37 patients with secondary amyloidosis entered dialysis treatment at our department. All had amyloidosis secondary to chronic arthritic disease (35) or to other chronic inflammatory causes (2). Only two patients were maintained on CAPD throughout follow-up; 12 patients (32%) received a kidney transplant. Survival in dialysis at 1 year was 82%, at 2 years 46%, and at 3 years 37%. Survival of amyloidosis patients transplanted at the Finnish transplant centre within the same period was worse at 1 year, but better at 2 and 3 years, 70%, 62%, and 62% respectively, but the difference was not significant. Populations are not compatible, since patients were selected for transplantation. Infection was a common cause of death, 7/18 (39%) deaths; cardiac deaths were less common, only two myocardial infarctions and one cardiac arrhythmia (17%). Symptoms of cardiac amyloid infiltration indicated a poor prognosis, although it did not necessarily predict death of a cardiac cause. Cardiac infiltration of amyloid was more common in autopsy than previously reported (10 of 13 patients), probably indicating longer duration of amyloidosis in patients treated with renal replacement therapy. Patients who died within follow-up had a shorter interval between the start of primary disease and the development of amyloidosis than those who survived, 11.8 versus 17.7 years (P = 0.041), and also a slightly shorter period between diagnosis of amyloidosis and start of dialysis, 3.0 versus 4.4 years (P = 0.129). This indicates that the rate of progression of amyloidosis determines the development of disease-associated complications, and fast progression may predict serious outcome.


[Uterine sarcoma. Clinical and pathological study of 25 cases]. The study reports on the follow-up in 25 cases of uterine sarcoma from 1979 to 1988. 56% of them are leiomyosarcoma, 28% mixed mullerian tumors and 16% sarcoma of the endometrial stroma. The women affected are mostly 40 to 70 years old; 80% of them are pluriparae, in menopause for 64% of them at the time of the diagnosis. Metrorrhagia is the initial symptom in 72% of the cases, 76% of which diagnosed after surgical intervention. The majority of the surgical interventions are integrated with chemo and/or radiotherapy. After three years 66.6% are surviving at the first stage, 33.4% at the second stage, none at the third stage.


Blood pressure during the interdialytic period in haemodialysis patients: estimation of representative blood pressure values. The estimation of representative blood pressure (BP) levels is difficult in haemodialysis (HD) patients as it is not known whether pre- or postdialytic blood pressure are predictive for the average interdialytic BP. Furthermore, the day-night BP rhythm can be disturbed in HD patients. Therefore, in this study, BP was measured during the interdialytic period using non-invasive ambulatory BP measurements in four hypotensive, six normotensive, and 12 hypertensive HD patients. It was assessed whether pre- or postdialytic BP was representative for the average interdialytic BP. Furthermore, the nocturnal BP reduction was compared between HD patients, seven normotensive controls and eight treated subjects with essential hypertension. Postdialytic BP was superior to predialytic BP in predicting the average BP during the interdialytic period. BP did not differ significantly between day 1 and day 2 of the interdialytic period but increased rapidly in the hours before dialysis. Weight gain (corrected for actual body-weight) did not correlate significantly with the increment in systolic BP (r = 0.21; P = 0.2) or diastolic BP (r = -0.02; P = 0.5) during the interdialytic period. The nocturnal decline in systolic BP was significantly attenuated (P less than 0.001) in hypertensive HD patients compared with normotensive controls. The nocturnal reduction in diastolic BP was significantly less in hypotensive (P less than 0.001) and normotensive (P less than 0.001) HD patients compared with normotensive controls and in hypertensive HD patients compared with normotensive (P less than 0.001) and hypertensive (P less than 0.001) controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Failure of a daily haemofiltration programme using a highly permeable membrane to return beta 2-microglobulin concentrations to normal in haemodialysis patients. In an attempt to return to normal serum beta 2-microglobulin levels in a group of seven ESRD patients, a programme of daily HF with highly permeable AN69 membranes was undertaken. Pre-HF beta 2-M serum levels stabilized after 4 days at 20 mg/l, only 40% lower than the initial concentration. A total of 985 +/- 20 mg beta 2-M was removed over the week. The beta 2-M release rate averaged 97 micrograms/min with a broad range of values (63-128 micrograms/min). beta 2-M release peaked at 602 micrograms/min 1 h after the end of the HF session before returning to baseline by 12 h post-HF. We conclude that a return to normal blood beta 2-M concentrations in ESRD patients seems quite unrealistic despite a highly intensive extracorporeal therapy. Therefore other therapeutic alternatives have to be designed to prevent or cure beta 2-M amyloidosis.


Pharmacokinetics of aluminoxamine and ferrioxamine and dose finding of desferrioxamine in haemodialysis patients. We investigated the pharmacokinetics of desferrioxamine and its chelated compounds aluminoxamine and ferrioxamine in normal volunteers and haemodialysis patients with and without iron overload. Desferrioxamine was administered in a single dose of 30 mg per kg body-weight was a 30-min infusion to five healthy volunteers and to 20 haemodialysis patients (five patients without haemosiderosis and 15 patients with haemosiderosis). The interdialytic half-life of ferrioxamine was 2.2 h in normal volunteers, 13.3 h in dialysis patients without haemosiderosis, and 24.6 h in patients with haemosiderosis. There was no interdialytic elimination of aluminoxamine. In a second study, seven dialysis patients received 5, 10, and 20 mg per kg body-weight desferrioxamine in a random order with a time interval of 2 weeks. The peak serum concentrations after these doses were 4.1 +/- 2.9, 6.4 +/- 2.9, and 10.7 +/- 7.1 mumol/l for ferrioxamine and 2.8 +/- 1.5, 3.1 +/- 1.5, and 4.2 +/- 1.7 mumol/l for aluminoxamine. Thus, a 4-fold increase in desferrioxamine dosage resulted in a 2.7-fold increase in peak ferrioxamine levels and in only a 1.5-fold increase in peak aluminoxamine levels. We conclude that dialysis patients, especially those with haemosiderosis, are exposed to persistently elevated ferrioxamine levels. Weekly doses of 5-10 mg/kg of desferrioxamine would be sufficient for aluminium chelation therapy.


Aluminium overload and response to recombinant human erythropoietin in patients under chronic haemodialysis. In this work, of 51 patients treated by rHuEpo, 25 were selected for study. The selection criteria were absence of clinically evident causes of anaemia other than end-stage renal failure, such as chronic infection, active systemic disease, bleeding sites, and vitamin B12 or iron deficiencies. Serum aluminum was assessed before dialysis and the presence of aluminium overload was confirmed by a DFO test. rHuEpo was given in a dose of 50 U/kg body-weight after each dialysis session three times weekly and the response to treatment was evaluated monthly for 8 months. Our data showed significant correlation between serum aluminum and the response to rHuEpo. The response was significantly greater in those with lower serum aluminium. We conclude that the aluminium load in chronic haemodialysis patients may have an effect on the response to rHuEpo.


Immunoadsorption of anti-HLA antibodies for highly sensitized patients awaiting renal transplantation. Fifteen end-stage renal disease patients with high titres of panel reactive (PRA) antibodies were treated with immunoadsorption (IA) on sepharose-bound protein A columns in order to remove anti-HLA antibodies and facilitate transplantation. Infectious complications were not observed after IA and transplantation, and the procedure was well tolerated. In spite of the use of adjunctive immunosuppressive treatment with cyclophosphamide and prednisolone, this method produced only variable effects in lowering panel reaction antibodies, and was hampered by high de novo resynthesis of anti-HLA antibodies. Patients whose pre-IA antibody titre was greater than or equal to 1:64 clearly did not benefit from the procedure, but other immunological criteria were not predictive of efficacy. Twelve patients were transplanted on the basis of a negative cross-match with current serum, historical sera being retrospectively tested. Surprisingly, seven patients received a well-matched graft with both pre- and post-IA negative cross-matching. Graft survival was 86% in this group. Conversely, in the group of five transplants which were performed in recipients having a positive historical cross-match with the donor, graft survival was only 40%. One patient died with a functional graft, and two grafts failed due to hyperacute humoral rejection. Humoral rejection in a third patient was successfully treated by a second IA course and administration of polyclonal IgG. We conclude that IA is a safe procedure for managing hyperimmunized transplant candidates. However, its efficacy remains variable, and a better definition of patients who should benefit from IA needs to be found.


Pigeons and peritonitis?

Pigeons and peritonitis? We report an outbreak of fungal peritonitis due to Candida parapsilosis in 12 patients undergoing chronic ambulatory peritoneal dialysis (CAPD). All 12 patients were treated by removal of the CAPD catheter together with systemic antifungal therapy. There were no peritonitis-related deaths. Four patients were successfully returned to CAPD at a later date. Microbiological investigation during the outbreak demonstrated colonization of various areas of the CAPD Unit and medical ward with the organism. C. parapsilosis was also isolated from pigeon guano obtained from window-sills. The number of cases of peritonitis due to this organism decreased markedly after bird-proof netting was installed. We believe that this is the first report of an outbreak of CAPD peritonitis due to faecal carriage of C. parapsilosis by pigeons.


Synaptic enhancement induced by NMDA and Qp receptors and presynaptic activity. Activity-dependent changes in synaptic efficacy may underlie the acquisition of memory. Much interest has centred on the involvement of NMDA (N-methyl-D-aspartate) receptors in the induction of long-term potentiation (LTP), but since activation of this receptor alone results only in short-term enhancement, other receptors are clearly involved. This paper describes a novel form of lasting synaptic enhancement requiring concomitant activation of both NMDA and metabotropic quisqualate, or Qp receptors for induction. Neither high frequency presynaptic activity, co-activation of many presynaptic axons, postsynaptic depolarization, nor blockade of inhibition were required. However, repetitive, though relatively low frequency activity of the presynaptic axon appeared to be necessary. We suggest, tentatively, that presynaptic activity may be directly involved in priming the recently active terminal to respond to postsynaptic changes.


Influence of Na/Ca exchange and mobilization of intracellular calcium on the time course of the slow afterhyperpolarization current (IAHP) in bullfrog sympathetic ganglion neurons. IAHP is a calcium dependent potassium current that underlies slow afterhyperpolarizations following action potentials in bullfrog sympathetic ganglion neurons. The decay rate of IAHP increases with increasing calcium loads. This effect was found not to be due to mobilization on intracellular calcium from ryanodine and caffeine sensitive stores. The relation is not affected by ryanodine at concentrations that block mobilization in the presence of caffeine, a drug that enhances mobilization of those stores. Nor does the relation seem to be due to a reduction of the driving force of the Na/Ca exchange process. The relation between decay rate and calcium load persists when Na+ is replaced by Li+. Our results suggest that Na/Ca exchange and mobilization of intracellular calcium normally have little influence in determining the time course of IAHP in these neurons.


Development of glycine receptor alpha subunit in cultivated rat spinal neurons: an immunocytochemical study. Ontogenesis of the inhibitory glycine receptor was studied up to 12 days in vitro in spinal neurons placed previously in culture at embryonic day 14. The alpha subunit of the receptor was detected using standard and confocal immunofluorescence and a specific monoclonal antibody. The immunostaining was compared to that of synaptophysin, a synaptic vesicle antigen, which was taken as an index of synaptic maturity. Glycine receptors could be detected intracellularly, and not at the cellular surface in some cells as early as 2-3 days in vitro (DIV) prior to any synaptic contact. At 4-5 DIV, the number of cells which expressed the immunoreactivity and the fluorescence intensity increased. At this stage, spherical fluorescent blobs started to migrate in the neurites. From 6 DIV, the glycine receptor alpha subunit was detected at the neuronal surface and was organized in clusters whose number increased progressively with time. From 7 DIV, the intrasomatic immunoreactivity decreased, and at day 12, the pattern of labelling was similar to that observed in the adult spinal cord. A diffuse presence of the receptor at the surface of neurons could never be visualised, and when detected, the glycine receptors were always clustered. Thus, the increasing expression of clusters of glycine receptors at the neuronal surface was paralleled by that of synaptophysin in neuritic varicosities. These data suggest that transport of glycine receptors to the plasmamembrane and the formation of aggregates occurs simultaneously to synaptogenesis.


Coordination of ATP production and consumption in brain: parallel regulation of cytochrome oxidase and Na+, K(+)-ATPase. Previous studies have shown that most neuronal ATP is produced by oxidative metabolism, and consumed by Na+,K(+)-ATPase. We hypothesized that the distribution of Na+,K(+)-ATPase in brain would correlate with that of the energy-producing enzyme, cytochrome oxidase (CO). We localized these enzymes in monkey hippocampus and striate cortex by histochemistry and immunohistochemistry. Their distributions were generally similar, although some differences were observed. We also studied regulation of enzyme levels, using monocular impulse blockage with tetrodotoxin (TTX) to alter neuronal activity in the visual system. Parallel changes in CO and Na+,K(+)-ATPase activity were induced in striate cortex. These results provide further evidence that neuronal energy demands regulate CO levels and distribution.


The ontogeny of 2-[125I]iodomelatonin binding sites in chicken brain. The characteristics of the binding sites for 2-[125I]iodomelatonin were studied in chicken brain membranes during development. Specific binding, defined using cold melatonin (1 microM), was detected as early as 8-day-old embryos. Scatchard analysis of saturation experiments showed that 2-[125I]iodomelatonin binds to a single class of site at all ages tested (8-day-old embryos to 3-month-old chicks). Binding affinity (Kd) did not change during development (18-31 pM), but the maximal number of binding sites (Bmax) increased until embryonic day 18, and then remained relatively constant until 30 days of age. A further increase in Bmax was seen at 3 months of age. Guanosine 5'-triphosphate (GTP, 1 mM) inhibited 2-[125I]iodomelatonin binding at all ages suggesting that the melatonin binding site is coupled to a guanine nucleotide binding protein at a very early stage of development. Competition experiments with a number of melatonin analogues indicated that the binding site detected in the brain at embryonic day 8 was pharmacologically identical to that observed 15 days after hatching.


Origin of brain 2',3'-cyclic-nucleotide 3'-phosphodiesterase doublet. The present study established that 2',3'-cyclic-nucleotide 3'-phosphodiesterase doublet common to mammalian brain originates from an alternative splicing. Peptides specific to the predicted larger translation product were synthesized and antisera against these peptides were prepared. Immunostaining of SDS/PAGE blots showed that the antisera react with the larger protein, but not with the smaller protein, of 2',3'-cyclic-nucleotide 3'-phosphodiesterase doublet in all mammals studied.


Comparison of carbon monoxide and nitrogen induced effects on synaptic transmission in the rat hippocampal slice. A comparison has been made of the effects of carbon monoxide (CO) or nitrogen (N2) exposure on synaptic transmission in the hippocampal slice. CA1 field potentials, evoked by Schaffer collateral stimulation, were unaffected by superfusion of slices with artificial cerebral spinal fluid (ACSF) equilibrated with either 15% CO or 15% N2 for 120 min. However, superfusion with hypoxic ACSF equilibrated with either 85% CO or 85% N2 caused a rapid depression of synaptic transmission. Reperfusion with control ACSF following 30 min hypoxia led to recovery of evoked responses and a slight hyperexcitability. In the hippocampal slice, synaptic transmission, as assessed by input/output curves, was not different during or following hypoxia induced by exposure to CO or N2. In the short term, CO is not toxic.


A third type of calcium current in cultured human skeletal muscle cells. A third type of calcium current could be recorded on a non-negligible number of human skeletal muscle cells (normal and Duchenne dystrophic (DMD)) in primary culture. This transient current exhibited a maximum at 0 mV, a time-to-peak around 30 ms, an inactivation time constant around 70 ms and was insensitive to nifedipine. On these basis, it differentiates from T- and L-type previously described and looks like the neuronal N-type. However, this third type of current was not sensitive to omega-Cgtx, a specific N-type blocker. The occurrence and the possible role of this current are briefly discussed.


Role of NK-1 receptor in central cardiovascular regulation in rats: studies on a novel non-peptide antagonist, CP-96,345, of substance P NK-1 receptor. CP-96,345[(2S,3S)-cis-2-(diphenylmethyl)-N-[(2-methoxyphenyl)- methyl]-1-azabicyclo [2.2.2] octan-3-amine] was recently discovered to be a nonpeptide substance P (SP) antagonist. We examined the effects of CP-96,345 on the central cardiovascular responses to tachykinin peptides in anesthetized rats. CP-96,345 (200 nmol, i.c.v.) inhibited the pressor responses of the NK-1 receptor-selective agonist GR 73632 (0.5 nmol, i.c.v.) and SP (7 nmol, i.c.v.). It also inhibited the increase in blood pressure elicited by neurokinin A (7 nmol, i.c.v.). However, it had no effect on the earlier pressor response induced by neuropeptide gamma (l nmol, i.c.v.) or by a selective NK-3 agonist senktide (1 nmol, i.c.v.). These findings suggest that SP (i.c.v.) induces pressor responses via the NK-1 receptor, and that the pressor response to neurokinin A may also be mediated by the NK-1 receptor in the brain.


Post-inhibitory excitation of adenosine on neurotransmission in guinea pig hippocampal slices. The postsynaptic field potential (population spike potential; PS) was recorded from the granule cell layer of guinea pig hippocampal slices. Adenosine at low concentrations ranging from 10 nM to 1 microM enhanced the amplitude of PS, whereas at concentrations over 10 microM it inhibited the neurotransmission. There appeared to be a rebound phenomenon after the removal of adenosine at inhibitory concentrations and the amplitude of the PS overshot the initial amplitude (we called this post-inhibitory excitation; PIE). Neither depressants such as gamma-aminobutyric acid (GABA; 1 mM) nor sodium pentobarbital (100 microM) by itself induced PIE. After application of GABA or sodium pentobarbital together with adenosine (0.1 microM), however, removal of all agents could induce the PIE. PIE as well as the excitatory effect of adenosine at low concentrations was counteracted by application of H-7 (100 microM), melittin or polymyxin B, potent protein kinase C (PKC) inhibitors, suggesting that the excitatory effect of adenosine is mediated by a metabolic process involving PKC. These results indicate that PIE induced by adenosine at high concentrations is due to a mechanism similar to the excitatory effect induced by adenosine at low concentrations, and that during application of adenosine at high concentrations the excitation is masked by its potent inhibitory effect.


Ubiquitin profile and amyloid enhancing factor activity in Alzheimer and 'normal' human brain extracts. Tris-HCl or Laemmli sample buffer extracted frontal lobe and hippocampal samples from normal aged and Alzheimer's disease (AD) subjects were used to determine total ubiquitin (Ub), distribution of monomeric Ub and Ub-protein conjugates and amyloid enhancing factor (AEF) activity using the dot-blot, Western blot and mouse AEF bioassay techniques, respectively. The AD samples, as compared to the normals, demonstrated a 1.7-fold increase in total Ub, elevated levels of Ub-protein conjugates and an appreciably enhanced AEF activity. Many of the hippocampal Ub-protein conjugates were found to be soluble only in the Laemmli sample buffer. The possible roles of elevated Ub levels and of the association of AEF activity with Ub are discussed in regard to pathogenesis of brain amyloidosis.


Marked increase in cerebrospinal fluid ubiquitin in Creutzfeldt-Jakob disease. We have established the radioimmunoassay for ubiquitin in the cerebrospinal fluid (CSF) and measured the ubiquitin concentration in CSF from 4 cases of neuropathologically verified Creutzfeldt-Jakob disease (CJD), 10 cases of multi-infarct dementia (MID), 7 cases of senile dementia of Alzheimer type (SDAT), and 18 controls. The normal values were determined to range from 7.3 to 21.0 ng/ml, 14.3 +/- 1.1 ng/ml in the mean +/- S.E.M. The CSF ubiquitin levels in the cases of MID and SDAT were 16.6 +/- 6.4 ng/ml and 21.3 +/- 6.1 ng/ml, respectively. In the cases of CJD, the CSF ubiquitin was markedly increased at the early and middle stages of the disease (230.6 ng/ml in Case 1, 107.6 ng/ml in Case 2, 212.5 ng/ml in Case 3, and 377.0 ng/ml in Case 4) and these gradually decreased as the disease progressed. The measurement of CSF ubiquitin seems useful to make an early diagnosis of CJD.


The facilitating and suppressing effects of delta 9-tetrahydrocannabinol on the rise in intrasynaptosomal Ca2+ concentration in rats. The effects of delta 9-tetrahydrocannabinol (delta 9-THC) on the rise in intracellular Ca2+ concentrations ([Ca2+]i) after stimulation with 15 mM or 29 mM K+ in rat whole brain synaptosomes were examined. A fluorescent chelating agent, Rhod-2, was employed to monitor any alterations of K(+)-evoked [Ca2+]i. Pretreatment with 10(-10) M delta 9-THC for 3 min enhanced K(+)-evoked [Ca2+]i significantly, while 10(-9), 10(-8) or 5 x 10(-8) M delta 9-THC significantly inhibited the K(+)-evoked [Ca2+]i. These results suggest that delta 9-THC had a biphasic effect on the K(+)-evoked Ca2+ response in rat brain synaptosomes.


Identification of a unique nuclear receptor for 9-cis retinoic acid. Two families of nuclear receptors (RAR and RXR) for retinoic acid have previously been identified, cloned, and shown to be intimately involved with receptor-mediated regulation of gene transcription. All-trans-retinoic acid (t-RA) has been known to be a high-affinity ligand for RAR but only a weak ligand for RXR, while the endogenous ligand for RXR was unknown. Now two laboratories have isolated and chemically characterized 9-cis-retinoic acid (c-RA) and have shown that it is a naturally occurring ligand for RXR. These results suggest that isomerization of t-RA into c-RA may play an important role in retinoic acid biology.


Sexually transmitted diseases: perspectives on this growing epidemic. Despite advances with antibiotic therapies and increased public attention to acquired immunodeficiency syndrome, sexually transmitted diseases remain a major public health problem of epidemic proportions in the United States. The emergence of new pathogens and clinical syndromes has increased the complexity of the situation. Incurable conditions and asymptomatic disease states during an era of constrained public resources and deficient public awareness further heighten and compound the devastating consequences of sexually transmitted disease. The provision of comprehensive disease prevention, education and clinical services is an important and complicated dimension. Sexually transmitted diseases are responsible for considerable morbidity and substantial mortality, particularly among young women. This article provides an overview of the epidemic in financial and human terms, with a discussion of eight major pathogens from an epidemiological, clinical and educational-counseling perspective.


A review of toxic shock syndrome: the need for education still exists. Toxic shock syndrome is a multisystem disease that is believed to be caused by a toxin produced by Staphylococcus aureus. The syndrome can occur in persons of any gender, age or race and should be considered as a possible diagnosis in any patient displaying symptoms compatible with the illness. Most reported cases of toxic shock syndrome have occurred in young menstruating women and have been associated with the use of tampons. Toxic shock syndrome cases reported to the Centers for Disease Control have decreased substantially since May of 1980. The decreases in diagnosis and reporting may be attributed to diminished public and professional attention to the syndrome. In addition, it is likely that cases of the syndrome that do not meet the strict Centers for Disease Control case definition are not being diagnosed. Until more is understood about toxic shock syndrome, particularly its relationship to menstruation and tampon use, patient education remains a primary key to prevention. Women should be aware of the symptoms of toxic shock syndrome and, if symptoms occur, seek medical attention immediately.


Bone marrow scintigraphy in lung carcinomas using nanosized colloids: when is it useful and how useful is it? Bone marrow scintigrams (MS) combined with single photon emission computed tomographic investigation of the liver (liver SPECT) were obtained using 99Tcm-labelled human serum albumin nanosized colloids in 52 patients with histologically proven lung carcinomas (adenocarcinomas = 17, squamous cell = 16, small cell = 14, large cell = 5, 31 generalized cases at the time of the first MS investigation among whom 13 patients had proven skeletal metastases). They were compared with conventional bone scintigrams (BS) as well as clinical, biological, radiological and follow-up data obtained for the same patients. In the present series, MS appeared as sensitive as BS in diagnosing skeletal metastases (77%) if all abnormal MS and BS presentations are considered as diagnostic, but more sensitive (77% versus 54%) if more restrictive analytical criteria are applied. The two investigations yielded the same specificities whatever the analytical criteria applied. These preliminary conclusions have, however, to be confirmed on larger populations than the present series. The most striking differences between BS and MS were observed in the case of small-cell lung carcinomas, with more lesions detected by MS than by BS. Liver SPECT also made it possible to diagnose seven and to suspect one out of the ten situations of hepatic metastases. The combination of liver spect and MS in the framework of a single injection of 99Tcm-labelled nanosized colloids thus allowed us to diagnose 80% of the patients with osseous and/or hepatic metastasis or 40% of all generalized cases.


Anti-beta HCG abdominopelvic immunoscintigraphy in patients with resistant gestational trophoblastic disease. The clinical utility of abdominopelvic immunoscintigraphy (IS) using anti-beta HCG-labelled monoclonal antibodies was evaluated in six patients with resistant gestational trophoblastic disease and no common pathological site. Five patients with abnormal retrovesical uptake had persistence of uterine trophoblastic disease confirmed on hysterectomy. Four of these patients are now in remission. It is concluded that IS provides a useful indication for surgery when a single abnormal uptake site is found.


Clinical outcomes of patients with suspected pulmonary embolism using 99Tcm-Technegas as a ventilatory agent for lung scanning. Clinical outcome analysis was carried out in 175 of 206 consecutive patients referred for a lung scan with clinical suspicion of pulmonary embolism (PE). The follow-up time period ranged from 4 to 18 months. High-quality ventilation images corresponding to the six standard perfusion images were obtained using Technegas as a ventilatory agent. Lung scan reports showed that 22% of the patients had a high, 14% indeterminate, 18% low and 9% very low probability for PE and 37% were normal lung scans. These reports usually supported the referring clinicians' provisional diagnosis and were confirmed by the clinical outcome analysis of these patients giving an apparent sensitivity for the lung scan of 96% and specificity 93%. A strategy for lung scanning in PE is proposed.


Mixed mesodermal tumor of the ovary: analysis of prognostic factors in 31 cases. To determine significant prognostic factors in patients with mixed mesodermal tumors of the ovary. Thirty-one cases of mixed mesodermal tumor of the ovary treated at Memorial Sloan-Kettering Cancer Center between 1977-1990 were reviewed retrospectively. The mean patient age was 61 years. Distribution by stage was as follows: I, seven (23%); II, one (3%); III, 15 (48%); and IV, eight (26%). The median follow-up for survivors was 62 months. Following primary surgery, chemotherapy included cisplatin (four), doxorubicin (seven), or both (ten); six patients received various other treatments. The median survival for the entire group was 10.6 months. In 19 cases (61%), heterologous sarcomatous elements were present in the primary tumor, whereas 12 (39%) contained homologous elements only. There was a trend toward improved survival in patients whose primary tumors had only homologous stromal elements (P = .06). The overall survival was significantly better for the eight patients with early-stage (I, II) disease than for the 23 patients with advanced-stage (III, IV) disease (P = .01). The size of residual disease after cytoreductive surgery was not a significant prognostic factor. There was no difference in survival between the ten women whose metastatic disease contained only epithelial elements and the 16 whose metastases contained mesenchymal elements as well (P = .23). This study confirms previous observations that mixed mesodermal tumors of the ovary are a highly malignant group of tumors that respond poorly to chemotherapy. In addition, we demonstrated that prognosis is independent of the presence or absence of sarcomatous elements in the metastases.


Invasive cervical cancer following cryotherapy for cervical intraepithelial neoplasia or human papillomavirus infection. To determine the following: 1) the causes for the failure of cervical cryotherapy to prevent cervical cancer, and 2) whether cervical cryotherapy is associated with the development of cervical adenocarcinoma rather than squamous carcinoma. We reviewed the medical charts of 327 women with cervical cancer. One hundred forty-seven for whom pertinent history was missing were contacted by telephone or at clinic visits. History obtained verbally was confirmed by outside medical records. Cervical biopsies (N = 16) and endocervical curettages (ECCs) (N = 15) performed before cryotherapy and biopsies at the diagnosis of cancer (N = 21) were reviewed. Twenty-one women with cervical cancer had a history of cryotherapy for cervical intraepithelial neoplasia (CIN) or human papillomavirus infection (HPV). The interval between cryotherapy and cancer was more than 2 years in 19 and more than 5 years in ten. Several categories of pre-treatment errors were identified. Evaluation before cryotherapy was appropriate in only nine cases. Interpretive errors were noted in three of 16 cervical biopsies and ten of 15 ECCs. After cryotherapy, 12% of women had appropriate follow-up. Of the invasive cancers that developed, 24% in the cryotherapy group and 21% in the non-cryotherapy group were adenocarcinomas. Careful evaluation before cryotherapy, accurate pathology reports, and consistent long-term follow-up are necessary if cryotherapy is to be used to treat CIN or HPV. We found no evidence that cryotherapy is associated with the development of cervical adenocarcinoma.


Etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine for the treatment of metastatic, high-risk gestational trophoblastic disease. To evaluate the efficacy and toxicity of a regimen of etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine in patients with metastatic, high-risk gestational trophoblastic tumors. Twelve women with metastatic gestational choriocarcinoma received 64 treatment cycles. All met the National Cancer Institute criteria for high-risk gestational trophoblastic tumors. Response was evaluated by monitoring serial serum beta-hCG levels. Toxicity was recorded using standard World Health Organization criteria. There was no life-threatening toxicity. Neutropenia necessitating a 1-week delay of treatment occurred with only eight treatment cycles (12.5%) and deferral of vincristine and cyclophosphamide with three cycles. Anemia requiring transfusion complicated only two cycles. Peripheral neuropathy in two patients was treated by discontinuing vincristine. Other toxicities included nausea and vomiting, diarrhea, stomatitis, alopecia, conjunctivitis, thrombocytopenia, and fever. Ten of the 12 subjects experienced a complete response. Two had partial responses and one with an initial complete response had relapse 4 months after completing therapy; all three were successfully salvaged with cisplatin-based chemotherapy. Overall survival was 100%, and all 12 patients are disease-free with a median follow-up of 26 months. Chemotherapy with etoposide, methotrexate, actinomycin D, cyclophosphamide, and vincristine is well tolerated and highly effective for metastatic, high-risk gestational trophoblastic disease.


Incidence of breast cancer in a 22-year study of women receiving estrogen-progestin replacement therapy. To review the incidence of breast cancer in a continuous 22-year study of conjugated estrogen-medroxyprogesterone acetate hormone replacement therapy. Eighty-four pairs of continuously hospitalized postmenopausal women who were matched for age, smoking history, and medical diagnosis were treated with estrogen-progestin hormone replacement therapy or placebo in a prospective and double-blind manner for 10 years. In the subsequent 12 years, the women were offered the choice of starting, stopping, or continuing hormone replacement therapy. After the initial 10 years, the incidence of breast cancer in the placebo group was 4.8%, whereas no cancers were found in the hormone replacement therapy group (P = .12). After an additional 12 years of follow-up, the overall incidence of breast cancer in the women who had never taken hormone replacement therapy was 11.5%, whereas no breast cancers had developed in the women who had ever taken hormone replacement therapy (P < .01). These data suggest that the 22-year administration of estrogen-progestin hormone replacement therapy did not increase the incidence of breast cancer in a small group of continuously hospitalized postmenopausal women.


A submicron emulsion as ocular vehicle for delta-8-tetrahydrocannabinol: effect on intraocular pressure in rabbits. delta 8-Tetrahydrocannabinol (delta 8-THC), a known antiglaucoma lipophilic drug, was incorporated in a submicron emulsion for ocular administration. The mean droplet size of the emulsion was 130 +/- 41 nm, and no droplet was larger than 400 nm. No change in pH, particle size distribution or zeta potential was noted after sterilization by steam autoclaving or long-term storage over 9 months. An intense and long-lasting intraocular pressure (IOP)-depressant effect was observed after ocular application (50 microliters) of the THC emulsion, 0.4% (w/w), to rabbits with ocular hypertension (chymotrypsin model). Lesser effects were observed in normotensive rabbits. No irritation effect of either the emulsion vehicle or THC emulsion on the rabbit eyes was detected. These results underline the promising properties of submicron emulsions as vehicles for lipophilic ophthalmic drugs. The mechanism by which the emulsion induced the marked delta 8-THC antiglaucoma effect remains unclear. However, the possible involvement of delta 8-THC systemic absorption in the hypotensive effect induced by the emulsion cannot be excluded and will be the subject of further investigation.


Atypical healing of cytomegalovirus retinitis. Significance of persistent border opacification. To analyze a phenomenon seen in patients with acquired immune deficiency syndrome (AIDS) with cytomegalovirus (CMV) retinitis undergoing systemic antiviral treatment: a persistent white border opacification on the edge of healed CMV retinitis. The authors prospectively evaluated a population of 137 patients with AIDS and CMV retinitis during a 44-month period. Eleven patients (12 eyes) who were undergoing maintenance antiviral treatment were identified with an atypical healing response--the persistence of a white flat border opacification that did not advance for many weeks to months. Patient records and photographs were reviewed. Results of one autopsy were analyzed with histopathology and special stains. The persistent white edge maintained (without advancement or smoldering) for an average of 11.6 weeks (range, 4 to 41 weeks). This border opacification was not affected by reinduction treatment in the six patients to whom reinduction was given. Results from histopathologic examination of one patient with a persistent white border are presented: these results show that dead cytomegalic cells formed stable structures within the retina, causing white opacification that could be confused with active lesions. Immunoperoxidase stains identified CMV antigens. This persistent white border opacification, which does not advance or smolder, represents an important clinical entity that should be recognized during antiviral treatment for CMV retinitis. It can often be observed. If it is not recognized as a stable configuration, patients may undergo unnecessary reinductions with potentially toxic doses of antiviral medications.


Visual field testing in the management of cytomegalovirus retinitis. Sequential visual field testing is an extremely helpful adjunct to ophthalmoscopy and fundus photography in the management of cytomegalovirus (CMV) retinitis with the antiviral agents ganciclovir or foscarnet in patients with the acquired immune deficiency syndrome (AIDS). The authors studied the visual field defects found in a series of 110 patients with AIDS and CMV retinitis. Ophthalmoscopy and fundus photography were performed on all patients. Visual field analysis was performed with either tangent screen, Goldmann kinetic, or Humphrey automated static perimetry. Of 166 eyes in 110 patients with CMV retinitis, visual field defects were present initially in 92 (55%) eyes of 78 (70%) patients, and ultimately in 97 (53%) eyes of 90 patients in whom follow-up was available. Stabilization of visual field defects was indicative of controlled retinitis. Sequential visual field testing will confirm ophthalmoscopic evidence of successful antiviral treatment of CMV retinitis and will corroborate very early progression of previously controlled retinitis.


Comparison of immunocytology to tissue culture for diagnosis of presumed herpesvirus dendritic epithelial keratitis. The objective of this study is to prospectively compare the sensitivity and specificity of immunodetection of herpes simplex virus (HSV) in impression cytology specimens obtained directly from presumed herpesvirus dendritic epithelial keratitis with virus isolation by tissue culture of cells scraped from the same lesion. Corneal impression cytology and tissue culture were performed on 29 consecutive patients presenting with presumed herpesvirus dendritic epithelial keratitis during a 6-month period. Impression cytology of dendritic epithelial keratitis lesions with Millipore Biopore membranes were evaluated for the presence of antigens specific to HSV type I (HSV-1), HSV-2, and varicella-zoster virus (VZV) using monoclonal antibodies specific to these herpesviruses and immunofluorescent staining techniques. Tissue culture was positive for HSV-1 in 52% (13 of 25) of dendritic epithelial keratitis patients without skin lesions, and was negative for VZV in 4 patients with dendritic epithelial keratitis and skin lesions in the distribution of the first division of the trigeminal nerve. The remaining 12 tissue cultures showed no cytopathic effect. Compared with tissue culture, impression cytology was 100% sensitive (13 of 13) and 92% specific (11 of 12) for the diagnosis of HSV-1 dendritic epithelial keratitis (Kappa coefficient of agreement 0.92). Although our sample size for VZV dendritic epithelial keratitis was small, the impression cytology findings correlated with our clinical diagnosis more often than tissue culture (2 of 4 versus 0 of 4). Impression cytology allows simultaneous debridement of dendritic epithelial keratitis and, when combined with immunocytologic staining procedures, provides a simpler, more rapid, and less expensive alternative to tissue culture for the diagnosis of dendritic epithelial keratitis caused by HSV or VZV.


Incidental oral hairy leukoplakia in immunocompetent persons. A report of two cases. In this brief article we report on two HIV-negative patients with documented oral hairy leukoplakia who had no known risk factors for HIV infection nor any evidence of other forms of immune suppression. Therefore we conclude that in some instances hairy leukoplakia can represent an isolated and innocuous Epstein-Barr virus infection.


The simultaneous occurrence of oral herpes simplex virus, cytomegalovirus, and histoplasmosis in an HIV-infected patient. Numerous publications have reviewed the oral manifestations of HIV infection and AIDS. Although uncommon, herpetic infections and histoplasmosis are among the diseases reported. The case that follows presents the first description of the simultaneous occurrence of oral herpes simplex virus, cytomegalovirus, and histoplasmosis in an HIV-infected person. These infections appeared as extensive oral ulcerations and were the only clinical manifestation of HIV infection. Eleven months after the initial presentation, the patient died of complications of AIDS.


Transformation of NIH3T3 cells by Rous sarcoma virus occurs with high efficiency in the absence of proviral rearrangements or amplification. NIH3T3 cells could be transformed by a mammaltropic strain of Rous sarcoma virus (RSV) with an efficiency 10(3) times greater than that observed in Balb/c 3T3 cells or other mammalian cell lines and almost identical to that of chick embryo fibroblasts. In infected NIH3T3 cells a single, properly integrated, provirus was sufficient to induce focus formation; moreover, kinase activity of pp60v-src and tyrosine phosphorylation of cellular proteins could be detected very soon after infection in the majority of cells. On the other hand, in transformed foci from RSV-infected Balb/c 3T3 cells both rearrangements and amplification of proviral sequences were frequently detected. Accordingly, expression of pp60v-src and ensuing tyrosine phosphorylation of cellular proteins occurred, at high levels, only in a minority of the infected cells. Furthermore, by using a murine retrovirus carrying the v-src oncogene and an independent selectable marker, we found that Balb/c 3T3 cells were transformed with a 100-fold lower efficiency than NIH3T3 cells, yet the majority of infected untransformed Balb/c 3T3 cells expressed active pp60v-src. These findings are consistent with the existence in most mammalian cell lines of a major restriction to v-src-induced transformation, operating at the level of proviral expression, that is apparently absent in NIH3T3 cells.


Autophosphorylation promotes complex formation of recombinant hepatocyte growth factor receptor with cytoplasmic effectors containing SH2 domains. Hepatocyte growth factor (HGF), also known as scatter factor (SF), is a polypeptide which induces motility and/or mitogenesis in epithelial cells. The receptor for HGF/SF, p190MET, is a two-chain transmembrane tyrosine kinase encoded by the MET proto-oncogene. To identify the cytoplasmic effectors involved in signal transduction we have produced the human HGF/SF receptor in insect cells (Sf9) by means of a recombinant baculovirus. Two 170-kDa forms of the receptor were synthesized in Sf9 cells: the uncleaved single-chain precursor (which is by far the more abundant) and the proteolytically processed two-chain molecule. Both receptor species are phosphorylated on tyrosine in vivo and are active kinases in vitro. The recombinant receptor binds and phosphorylates in vitro four known cytoplasmic transducers containing src homology region 2 (SH2) domains: the 85-kDa subunit of phosphatidylinositol 3-kinase (Pl 3-kinase), rasGAP, phospholipase-C gamma (PLC-gamma), and p59Fyn, a tyrosine kinase of the src family. In all cases the association is strictly dependent on tyrosine phosphorylation of the receptor, indicating that it occurs via specific interaction with the SH2 domains. These results show that the HGF/SF receptor has the sequence requirements for binding a spectrum of cytoplasmic transducers whose different combinations in target cells may result in the observed pleiotropic biological response.


Separation of immortalization from tumor induction with polyoma large T mutants that fail to bind the retinoblastoma gene product. The mouse polyomavirus encodes a tumor-suppressor gene inactivator in its large T protein and a proto-oncogene activator in its middle T protein. We have used site-directed mutagenesis to selectively inactivate the former function without affecting the latter. Two mutant viruses were constructed to encode altered large T proteins that fail to bind the retinoblastoma tumor-suppressor gene product pRB, along with normal small and middle T proteins. The pRB-binding mutants proved to be defective in immortalization of primary rat embryo fibroblasts by a variety of tests. Yet they proved capable of transforming both primary and established fibroblasts in culture. Most importantly, the inability of these mutants to bind pRB had little effect on their ability to induce tumors in mice. We conclude that induction of multiple tumor types in this system does not depend on large T-pRB interactions but rather on middle T-dependent pathways. In addition, the ability of this virus to immortalize cells in culture is not essential to its ability to induce tumors in the animal.