Cyclic AMP upregulates mRNA and surface expression of IL-2R alpha p55(Tac) in normal human NK cell clones. The effects of cAMP upon cell proliferation, cytotoxic activity, and regulation of IL-2R expression was investigated in normal human, IL-2-dependent natural killer (NK) cell clones. We report here that addition to the cultures of Bt2cAMP, a cell permeant analogue of cAMP, results in inhibition of IL-2-dependent proliferation, as assessed by [3H]Thymidine incorporation, in both NK and T cell clones. In addition, Bt2cAMP was shown to block the cytotoxic activity of NK cell clones at the level of the lytic phase. Contrasting with these inhibitory effects, cAMP induces an upregulation of the membrane expression of the IL-2R alpha chain (p55, Tac) in normal NK cell clones, which correlates with an accumulation of Tac mRNA. This is clearly at variance with T cell clones in which no such effect of cAMP alone is observed. In both cell types however, cAMP appears to synergize with IL-2 to increase IL-2R alpha mRNA expression. In addition, we demonstrate, using a cDNA probe to the IL-2R beta, that expression of this second component of the high affinity IL-2R, does not appear to be co-regulated together with IL-2R alpha in response to cAMP or/and IL-2 in cultured NK cells. Thus the effects of cAMP on human NK cell clones are complex. cAMP is inhibitory of proliferation and cytolytic function, whereas it is stimulatory of IL-2R alpha expression in these cells.


Cerebrospinal fluid antibodies to coronavirus in patients with Parkinson's disease. The etiology of Parkinson's disease remains unknown, and a search for environmental agents continues. In 1985, Fishman induced infection of the basal ganglia by a coronavirus in mice. Although coronavirus is recognized primarily as a respiratory pathogen in humans, its affinity for the basal ganglia led us to investigate its possible role in human Parkinson's disease. The cerebrospinal fluid of normal controls (CTL) (n = 18), and patients with Parkinson's disease (PD (n = 20) and other neurological disease (OND) (n = 29) was analyzed in a blinded manner by enzyme-linked immunosorbent assay [measurements in optical density (OD) units] for antibody response to four coronavirus antigens: mouse hepatitis virus JHM (J) and A59 (A), and human coronavirus 229E (E) and OC43 (O). When compared with CTL, PD patients had an elevated (p less than 0.05) mean OD response to J (0.0856 vs. 0.0207) and A (0.1722 vs. 0.0636). Response (p greater than 0.05) to O (0.0839 vs. 0.0071) was greater than that to E (0.1261 vs. 0.0743). When compared to OND, PD patients had an elevated mean OD response to J (0.0856 vs. 0.0267, p less than 0.05). Responses (p greater than 0.05) to A (0.1722 vs. 0.0929) and O (0.0839 vs. 0.0446) were greater than that to E (0.1261 vs. 0.0946). These results suggest that there may be an association between coronavirus and PD.


AAEM minimonograph #38: neuropathies in connective tissue disease. Neuropathies are common in patients with known or suspected connective tissue disease. A vasculitic mononeuropathy multiplex is often seen in patients initially presenting with polyarteritis nodosa or developing arteritis as a complication of rheumatoid arthritis. However, vasculitic neuropathy may become confluent and present as as distal symmetrical polyneuropathy or occur without systemic necrotizing vasculitis. Distal symmetrical polyneuropathies without associated vasculitis are also common in many connective tissue diseases. Compression neuropathies, especially carpal tunnel syndrome, occur with increased frequency in rheumatoid arthritis. Finally, certain neuropathies may be the major presenting feature of particular connective tissue diseases. For example, trigeminal neuropathy often heralds the onset of systemic sclerosis or mixed connective tissue disease, and sensory neuronopathy may be the initial presenting feature of Sjögren's syndrome.


Sural nerve biopsies in Guillain-Barre syndrome: axonal degeneration and macrophage-associated demyelination and absence of cytomegalovirus genome. T-cell infiltration was detected by immunohistochemistry in only 2 of 10 sural nerve biopsies from patients with Guillain-Barré syndrome (GBS). The number of endoneurial macrophages, identified by the monoclonal antibody MAC 387, was increased, compared with the number in 10 cases of axonal neuropathy. Macrophage-associated demyelination was identified in 7 and axonal degeneration in 8 cases. Cytomegalovirus (CMV) genome was not detected with the polymerase chain reaction.


Determining neuromuscular jitter using a monopolar electrode. Neuromuscular jitter was determined in the extensor digitorum communis (EDC) of 41 healthy control subjects and 8 patients with myasthenia gravis (MG). Standard single-fiber electromyographic (SFEMG) techniques were used, except that a monopolar electrode (MPE) was substituted for a single-fiber electrode (SFE). In normals, mean jitter for 20 pairs was 22.4 +/- 2.8 microseconds and the mean jitter for an individual pair was 22 +/- 5.6 microseconds. In the age range tested (18 to 49 years), jitter did not change with increasing age. When MPE jitter studies were repeated in 16 normals, mean jitter usually varied by less than 10% (mean 7.4% +/- 6.2%) and remained normal. Jitter was significantly increased in MG compared to normals 73 +/- 56 microseconds versus 22 +/- 5.6 microseconds (F = 149.4, P less than 0.0001). Using an analog pain scale, the discomfort during electromyography using MPE and SFE was compared between the left and right arms in 35 normals. MPEs caused less discomfort than SFEs (P = 0.0031), and were preferred (71% versus 20%) to SFEs (chi 2 = 9.21, P = 0.01). Based on these results, we conclude that MPE determined jitter studies are reproducible, can distinguish between normals and MG, and are associated with less discomfort than SFE.


[Viscerosomatic convergence on the lumbar interneurons of the dorsal horn of the spinal cord in cats and rats]. It is shown in experiments on spinalized cats under electrical stimulation of the pelvic nerve branch innervating the rectum as well as of the fibular nerve that 12 of 30 studied neurons to which the effect of these sources was converged were activated both by A- and by C-fibers. Reactions on stimulation of the fibular nerve were, apparently, mediated by the fibers with the conduction velocity not less than 2.2 m/sec. It is detected in experiments on spinalized rats that tension of the lower sections of the large intestine may evoke activation of neurons of the IV-V layers and inhibition of cells from the deeper layers. The inhibition evoked by the visceral stimulation in seven cases from 18 was provided by the effect directly on the postsynaptic membrane of these cells, in 11 cases it was localized in presynaptic (relative to them) structures. Naloxone, strychnine and atropine did not eliminate this inhibition which testifies against possible participation of opioids, glycines and acetylcholine in its creation. phaclofen, a blocker of the GABAB-receptors was also uneffective while bicuculline suppressed this inhibition in three cases from 12 which testifies to a probability of its participation in creation of GABAA-receptors.


[Relationship between the crystal lattice structure and the biological action of some agonists of amino acid receptors]. The crystal structures of glycine, taurine, GABA, beta-alanine were compared. The quantity and the accuracy of distances coincidence between nitrogen and oxygen atoms were used as a criterion of similarity of the crystalline structures. The conclusion is made about a correlation between crystalline structure of agonists and their effect on amino acid receptors. It is assumed that in case of a cooperative effect of agonist on the receptor a mutual arrangement of molecules on the receptor surface is similar to their arrangement in the agonist crystal.


[Mechanisms of calcium current decay acceleration induced by cyclic AMP]. In isolated snail neurones the level of cyclic AMP was increased either by intracellular injection of cAMP or by extracellular application of dibutyryl-cAMP and the change of high-threshold calcium current (ICa) decay was investigated. In 20 from 38 neurones it was obtained that both fast and slow phases of ICa decay were accelerated 2-2.5 times as affected by cAMP. The effect did not depend on the test-pulse potential and was displayed on the IBa. In the double-pulse experiments it was shown that cAMP enhanced the influence of depolarized prepulses (Vc) on the ICa tested (It). Analysis of the It-Vc curve showed that cAMP enhanced both Ca(2+)-dependent and voltage-dependent inactivation of ICa. The experiments where the intervals between Vc and Vt varied have shown that cAMP slowed down the rate of Ca(2+)-channels recovery from inactivation. The results suggest that cAMP increases the affinity of the Ca(2+)-channel inactivating substrate for Ca(2+)-ions.


Increased Na-K transport in glomerular mesangial cell membrane from spontaneously hypertensive rats. To investigate the differences in the Na-K transport of the mesangial cell (MC) membrane in hypertension versus normotension, the activity of the Na-K pump and the passive cation permeability were measured in serially passaged cultured MC obtained from both spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. When Na-K pump was active, Na-K pump activity, described as ouabain-sensitive 86Rb uptake, was significantly greater in the cultured MC from SHR than WKY rats. The outward Na-K cotransport, described as the washout rate constant of bumetanide-sensitive 86Rb washout, was also greater in SHR MC than in WKY rat MC. When Na-K pump was inhibited by 1 mM ouabain, overall intracellular Na uptake was significantly greater in SHR MC. A greater 5-(N,N-hexamethylene)amiloride-sensitive Na uptake in SHR MC accounted for this difference. There was no difference in the intracellular concentration of Na and K in the cultured MC from the 2 strains when Na-K pump was active. It is concluded that there is an increased activity of Na-K pump in the cultured MC from SHR, and that this abnormality may be innate to SHR cells. It is also suggested that an increase in Na-K cotransport and Na-H antiport may explain this difference, and that these abnormalities observed in the SHR kidney may be involved in the pathogenesis of hypertension in this model.


Acute effect of passive Heymann nephritis on renal blood flow and glomerular filtration rate in the rat: role of the anaphylatoxin C5a and the alpha-adrenergic nervous system. In earlier studies, we have shown that induction of passive Heymann nephritis (PHN) by intrarenal infusion of anti-Fx1A antibodies provokes an immediate fall in renal blood flow (RBF) and glomerular filtration rate (GFR). This was probably mediated via the complement system, as infusion of the F(ab')2 fraction of anti-Fx1A did not reduce RBF and GFR. In the present study, the effects of alpha-adrenergic blockade upon the acute hemodynamic changes during induction of PHN and of C5a infusion were studied. Group 1 was infused with anti-Fx1A antibodies during blockade of the sympathetic nervous system with the alpha-blocker phentolamine; control animals were treated similarly, but infused with normal rat IgG. Group 2 was infused with the anaphylatoxin C5a, normally produced during complement activation, and compared with control animals infused with saline. In group 1, RBF did not differ from control animals after the infusion of anti-Fx1A antibodies (6.6 +/- 0.5 compared to 7.3 +/- 1.0 ml/min/g in the controls). GFR in the left, antibody-infused kidney fell compared to controls, and was 0.25 +/- 0.08 ml/min/g at the end of the experiment compared to 0.60 +/- 0.13 ml/min/g (p less than 0.05 with Student's t test, p = 0.07 with two-way analysis of variance (ANOVA). GFR in the right kidney remained unchanged compared to controls. In group 2, C5a induced a significant fall in RBF (from 7.9 +/- 0.9 to 3.1 +/- 0.4 ml/min/g kidney weight), significantly different from control animals where it fell from 8.1 +/- 0.5 to 6.8 +/- 0.7 ml/min/g (p less than 0.0001 with two-way ANOVA, p less than 0.001 with t test).(ABSTRACT TRUNCATED AT 250 WORDS)


Pineal muscarinic phosphoinositide responses: age-associated sensitization, agonist-induced desensitization and increase in melatonin release from cultured pineal glands. Regulation of phosphoinositide (PI) signaling through the muscarinic cholinergic receptors (mAChRs) and their possible role were explored in the rat pineal gland. A sensitization of the PI signaling pathway was seen with advancing age. Binding of the mAChR ligand [N-methyl-3H]scopolamine to pineal sections, as detected by autoradiography, significantly decreased with advancing age and thus negatively correlated with the gland's ability to respond to cholinergic stimulus. The cholinergic agonist carbachol induced a time-dependent desensitization of the muscarinic PI signaling after 2 h of pretreatment in vitro (43 and 61% dampening of the PI response after 2 and 11 h pretreatment, respectively). This homologous desensitization was not mimicked by forskolin or phorbol esters, suggesting that proteins kinases A and C were not involved. Carbachol stimulation of the pineal glands in vitro increased melatonin release 2-fold, an effect quantitatively similar to that seen after adenylyl cyclase activation. Carbachol failed, however, to affect pineal cAMP levels. These results suggest that the PI signaling through pineal mAChRs is desensitized in young rats, possibly due to higher exposure to endogenous acetylcholine. Thus acetylcholine might play a prominent role in the developing gland. Moreover, acetylcholine could modulate melatonin release from the adult pineal gland in vivo.


Localization of 2-[125I]iodomelatonin binding sites in the brain of the Atlantic salmon, Salmo salar L. The photosensory pineal organ of teleost fish shows a circadian rhythm in melatonin synthesis, and melatonin is known to influence a number of physiological functions. However, the target sites for melatonin are not known. We have investigated the distribution of melatonin binding sites in the brain of the salmon, Salmo salar. Brains were collected for receptor binding assay and autoradiography at each of three time points: just after lights on, just before lights off, and in the dark at midnight (photoperiod light-dark 12:12, lights on at 08.00 h, lights off at 20.00 h). Specific binding of 2-[125I]iodomelatonin was observed in several brain areas. High densities were associated with (1) the optic tectum, (2) the preoptic area, (3) an area encompassing the magnocellular superficial pretectal nucleus ('nucleus rotundus') and the glomerular complex, (4) the inferior lobes of the hypothalamus, (5) the lateral mesencephalic tegmentum including the torus semicircularis, and (6) the molecular layer of the cerebellum. No binding was observed in the pineal organ or in the pituitary. We observed no differences in labeling between brains collected at different time points, except in the preoptic area where binding was high at 20.00 and 24.00 h, but low at 08.00 h, and in the corpus cerebelli, where labeling in the molecular layer was higher at 24.00 and 08.00 h than at 20.00 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Differential inhibition of dopamine and bromocriptine on induced prolactin release: multiple sites for the inhibition of dopamine. Effects of dopamine and bromocriptine on TRH- or dibutyryladenosine 3',5'-cyclic monophosphate (dbcAMP)-induced prolactin release from primary cultured rat pituitary cells were studied using a perifusion system. TRH (100 nmol/l) stimulated prolactin release from basal concentrations of 33.8 +/- 0.5 to 151.2 +/- 28.0 ng/ml (net increase) or 447% increase. Dopamine inhibited the basal release of prolactin throughout the experiment, but TRH (100 nmol/l) was still able to stimulate prolactin release under the influence of dopamine. The increment in prolactin release was inversely proportional to the dopamine concentration. When TRH (100 nmol/l) was introduced during a perifusion period with bromocriptine 1 nmol/l, the prolactin concentration was increased to 110.9% of basal levels. The stimulatory effect of TRH under the influence of bromocriptine (1 nmol/l) was significantly lower than that without bromocriptine (control), although the higher concentrations of bromocriptine (10 and 100 nmol/l) did not further reduce the peak concentration of TRH-induced prolactin release. During a perifusion period with a low concentration of dopamine (1 nmol/l plus 0.1 mmol/l ascorbic acid), introduction of dbcAMP (3 mmol/l) stimulated prolactin release to 48% of basal concentration. A higher concentration of dopamine further reduced the stimulatory effect of prolactin release. Bromocriptine impeded the stimulatory effect of dbcAMP (3 mmol/l) on prolactin release in a similar manner as dopamine. Since a higher concentration of bromocriptine (10 and 100 nmol/l) did not further inhibit the TRH-induced prolactin release whereas a higher concentration of dopamine did, it is concluded that dopamine acts through additional mechanism(s) other than the D2 receptor transduction system.


Phase I study of intracarotid administration of carboplatin. Fifteen patients were treated in a Phase I study of intracarotid carboplatin (200-400 mg/m2) in 5% dextrose and water infused over 15 to 30 minutes through a transfemoral catheter with a 0.2-micron inline filter. This study was done because intravenous carboplatin has less neurotoxicity than cisplatin and is active against brain tumors. Eleven men and four women ranging in age from 37 to 72 years (median, 59 years) were treated. The Eastern Cooperative Oncology Group performance status was 1 in 3, 2 in 4, and 3-4 in 8 patients. Eight patients had one to three previous chemotherapy regimens; previous radiotherapy had failed in 13 patients. The response of patients in the Phase I study follows: glioblastoma, 6 failed; not evaluated because of early death from pulmonary embolus, 1; recurrent Grade II and III glioma, 1 stable (minor response with neurologic improvement) and 2 failed; malignant oligodendroglioma, 1 failed; brain metastases from nonsmall cell lung cancer, 1 partial remission, 1 stable (minor response), and 1 failed; brain metastases from unknown primary, 1 stable (minor response with neurological improvement). Median survival was 9 weeks. Nausea was mild to moderate. One patient had granulocytopenia, and 2 had thrombocytopenia (mild). At 200 mg/m2 (2 patients), 1 had a focal seizure. At 300 mg/m2 (9 patients), 2 with abnormally small arteries had severe pain early in the treatment and posttreatment ipsilateral conjunctival edema, decreased vision, and cerebral edema (with partially reversible increased hemiparesis); 1 other had mild decrease in ipsilateral vision and 1 had transient aphasia on removal of the catheter (possibly the result of a vascular spasm).(ABSTRACT TRUNCATED AT 250 WORDS)


Local cerebral blood flow mapping before and after removal of acute subdural hematoma in the rat. The cause of hemispheric swelling and high intracranial pressure after acute subdural hematoma is unknown, yet this is a major cause of death related to this condition. To investigate whether vascular engorgement is the cause of this form of hemisphere swelling, we have autoradiographically mapped regional cerebral blood flow before and after removal of acute subdural hematoma in a rat model. Hyperemia was patchy and infrequent. The major cause of the significant hemisphere swelling seen after hematoma removal was enlargement of the zone of focal tissue ischemia and edema under the hematoma.


Effects of intracellular calcium chelation on voltage-dependent and calcium-dependent currents in cat neocortical neurons. Large neurons from layer V in a slice preparation of cat sensorimotor cortex were impaled with microelectrodes containing KCl plus different concentrations of the Ca2+ chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid (BAPTA) or two of its derivatives. Impalement with electrodes containing high BAPTA (200 mM) quickly abolished Ca(2+)-dependent afterhyperpolarizations. Spike parameters were normal, but the usual time- and voltage-dependent rectification of subthreshold membrane potential was absent. Normally, this rectification results from activation of two voltage-gated currents, the persistent sodium current (INaP) and the hyperpolarizing inward rectifier current (Ih). Both of these currents were absent during voltage clamp with high BAPTA microelectrodes. Impalement with electrodes containing low BAPTA (2 mM) or derivatives caused a different effect. Injection of a 1-s current pulse evoked phasic firing instead of the tonic firing seen normally. Both the amplitude and the duration of the Ca(2+)-dependent afterhypolarization that followed repetitive firing were much greater than normal. The effectiveness of BAPTA derivatives in altering afterhyperpolarizations and firing properties were similar to their effectiveness in chelating Ca2+. It is assumed that the BAPTA effects result from reduction of intracellular Ca2+ concentration. Results with high BAPTA suggest that (i) both INaP and Ih require a minimal intracellular calcium concentration for normal expression, and that (ii) these voltage-gated currents may be modulated by changes in intracellular calcium concentration. Results with low BAPTA suggest that a small reduction of intracellular calcium concentration preferentially enhances a slow, Ca(2+)-dependent K+ current which then dominates the firing properties of the cell. The transformed firing properties resemble those of hippocampal pyramidal neurons.(ABSTRACT TRUNCATED AT 250 WORDS)


Developmental and regional differences in the vulnerability of rat hippocampal slices to lack of glucose. Field excitatory postsynaptic potentials were recorded in stratum radiatum of CA1 and CA3 in submerged hippocampal slices from adult or newborn (postnatal days 5-25) Wistar rats. In adult slices, excitatory postsynaptic potentials were depressed by glucose removal ("aglycemia") more rapidly and to a greater extent in CA1 than in CA3 [respective mean times to 50% reduction in peak amplitude were 7.5 +/- 0.83 (standard error) min and 12.5 +/- 0.27 (standard error) min]. Subsequent recovery of excitatory postsynaptic potentials in normoglycemic medium was correspondingly quicker in CA3 than in CA1. Transmission failure at the synapses was indicated by the preservation of the afferent volley, and sharp depression of synaptic input-output plots. In the early postnatal period, CA1 excitatory postsynaptic potentials were much more resistant to aglycemia, substantially persisting for as long as 75 min, with full subsequent recovery in normoglycemic medium. The higher resistance of slices from newborn rats progressively disappeared over the first two postnatal weeks.


Simultaneous measurement of oxygen and dopamine: coupling of oxygen consumption and neurotransmission. Fast-scan cyclic voltammetry was used to simultaneously measure increases in dopamine concentration and decreases in O2 concentration evoked by brief electrical stimulation (two pulses at 10 Hz) in slices of rat caudate nucleus. Dopamine concentration began increasing immediately after the first pulse and reached a maximum within 200 ms of stimulation. The O2 concentration began to decrease 300-700 ms after onset of stimulus. Responses for both dopamine and O2 were dependent on external Ca2+ and were Cd2+ and tetrodotoxin sensitive. Only the O2 response was sensitive to CN- (0.15 mM). At short times after exposure to 50 microM ouabain, electrically stimulated dopamine overflow was increased by 150% and electrically stimulated changes in O2 concentration were unaffected. Maximum dopamine concentration was increased 28% by sulpiride (2 microM), 78% by L-DOPA (60 microM), 105% by nomifensine (10 microM) and unaffected by nialamide (10 microM). Maximum decrease in O2 concentration was increased by 25% by sulpiride and unaffected by nialamide, L-DOPA, or nomifensine. The decreases in O2 concentration are indicative of increased O2 consumption and are a measure of oxidative energy production evoked by electrical stimulation. The increase in dopamine is due to the release of dopamine balanced by uptake and serves as an indication of neurotransmitter activity. The results indicate that increases in oxidative energy production following electrical stimulation are dependent on external Ca2+ entry through Cd(2+)-sensitive channels. Possible mechanisms for this coupling are discussed.


Different postnatal development of cells expressing mRNA encoding neurotensin receptor. In situ hybridization histochemistry revealed three different ontogenetic patterns of localized expression of the high-affinity type of neurotensin receptor mRNA in the developing rat brain: one comprises sites which showed transient expression of neurotensin receptor mRNA during the first postnatal week, the expression greatly decreasing thereafter (type I); another comprises sites at which there is a gradual increase in neurotensin receptor mRNA after birth, as there is in cell number and intensity, with advancing age, followed by a plateau (type II); the third comprises sites at which there is much expression of neurotensin receptor mRNA already at birth, and a slight decrease thereafter (type III). The cerebral cortex, except retrosplenial and entorhinal cortices, and the anterior dorsal thalamic nucleus exhibit the type I pattern, while the horizontal and vertical limbs of the diagonal band of Broca, magnocellular preoptic nucleus, substantia innominata, ventral part of the suprachiasmatic nucleus, medial habenular nucleus, ventral tegmental area and substantia nigra pars compacta exhibit the type II pattern. The tenia tecta, retrosplenial and entorhinal cortices exhibit the type III pattern. One of the most striking findings in this study was that the entire neocortex and most of the limbic cortex exhibit the type I pattern, i.e. neurotensin receptor mRNA is expressed transiently long before a neuronal network is established there. This suggests that neurotensin plays an important role in cortical development, other than its reported transmitter-like role in the adult.


Effect of the GABAB antagonist, phaclofen, on baclofen-induced inhibition of micturition reflex in urethane-anesthetized rats. The effect of intrathecal or intracerebroventricular administration of the GABAB receptor agonist, baclofen, on rhythmic contractions induced by distension of the urinary bladder (micturition reflex) was evaluated in urethane-anesthetized rats. Baclofen inhibited bladder motility acting at central nervous system sites (spinal and supraspinal) with a comparable potency. The inhibitory effect of i.t. baclofen (0.1-10 nmol) was blocked by i.t. phaclofen (200 nmol) while i.c.v. phaclofen did not affect i.c.v. baclofen (0.1-1 nmol). The inhibition of the micturition reflex induced by bladder distension observed after i.t. administration of baclofen was unaffected by systemic capsaicin pretreatment (50 mg/kg s.c., four days before). On the other hand, i.t. baclofen suppressed, in a phaclofen-sensitive manner, the reflex bladder contraction evoked by chemical stimulation (topical capsaicin) of capsaicin-sensitive bladder afferents. Intrathecal baclofen did not affect the hexamethonium-resistant tonic contraction produced by topical application of capsaicin on to the urinary bladder, which is ascribable to local peptide release from sensory nerves. Bladder motility inhibition by i.t. or i.c.v. baclofen (1 nmol) was unchanged by previous administration of p-chlorophenylalanine, indicating that the serotonergic pathways do not play a role in its action. Baclofen (100 microM) suppressed the release of calcitonin gene-related peptide-like immunoreactivity evoked by electrical field stimulation from the dorsal half of the rat spinal cord. This response was also abolished by in vitro capsaicin desensitization or tetrodotoxin, indicating that baclofen suppresses transmitter release from central endings of capsaicin-sensitive primary afferents. The present findings indicate that baclofen acts at both spinal and supraspinal sites to inhibit, with different mechanisms, the micturition reflex.(ABSTRACT TRUNCATED AT 250 WORDS)


Involvement of intrathalamic GABAB neurotransmission in the control of absence seizures in the rat. The role of intrathalamic GABAB neurotransmission in the control of absence seizures was investigated. In rats with genetic absence epilepsy, bilateral injections of R-baclofen (50, 100 and 200 ng/side), a selective GABAB receptor agonist, into the specific relay nuclei and the reticular nuclei of the thalamus increased spontaneous spike and wave discharges in a dose-dependent fashion, whereas injections of a GABAB antagonist CGP 35,348 (1, 2.5 and 5 micrograms/side) into the same sites decreased these seizures dose-dependently. The effect of R-baclofen (200 ng/side) on spike and wave discharges could be blocked by a subsequent injection of CGP 35,348 (1 microgram/side) at the same site. Injections of R-baclofen (200 ng) or CGP 35,348 (5 micrograms) into the midline thalamus had no effect on these seizures. In non-epileptic rats, bilateral injections of R-baclofen (1 microgram/side) into the specific relay nuclei induced synchronized rhythmic oscillations on the cortical electroencephalogram. The results suggest that GABAB receptors in the ventrolateral thalamus and in the reticular nuclei are involved in an oscillatory activity which underlies the rhythmic spike and wave discharges recorded during spontaneous generalized non-convulsive seizures.


Grading of brain tumours. The British experience. In order to ascertain how widely the Kernohan and WHO numerical grading systems for brain tumours are used in Britain and how uniformly they are applied, a questionnaire was circulated to named neuropathologists in 19 Regional Neuropathology Centres in Britain. Eighty four per cent replied within 10 days. Their replies showed considerable variation in the use of grading systems and revealed that numerical grading was required mainly by clinicians and was not necessarily favoured by pathologists. Although some advantages of a grading system were advanced, it does seem that numerical grading of brain tumours is ill-defined and controversial. Furthermore, the use of historical grading systems may inhibit the incorporation of advances in pathological investigation into the clinical management of patients with brain tumours.


[Beta interferon therapy of HPV infections of the female genital tract]. The aim of the study was to evaluate the incidence of cervical and vaginal lesions due to Papilloma virus before and after beta-interferon therapy. Clinical symptoms and the therapeutic approach are described, whereas drug efficacy was assessed on a long-term basis. The study of 19 cases, brought to the Author's attention over an eight-year period (1984-91) confirms previously published reports: the use of interferon shows a good level of tolerability without complications and undeniable efficacy following surgical treatment.


[Efficacy of natural alpha interferon from normal human leukocytes in symptomatic vestibular papillomatosis]. The paper discusses the genesis of vestibular papillomatosis: some researchers affirm that it is epithelio-paraphysiological whereas others maintain that it is a subclinical stage of HPV infection. In some cases "true" symptomatic papillomatosis may be remedied by human leucocyte alpha-IFN treatment.


Role of vascular congestion in cisplatin-induced acute renal failure in the rat. The significance of vascular congestion in the pathogenesis of cisplatin acute renal failure (ARF) was studied in rats given pentoxifylline. Rats were administered single intraperitoneal doses of 1.0, 2.5, 5.0, and 10.0 mg/kg of cisplatin with 45 mg/kg of pentoxifylline or saline every 12 h for 3 days. Cisplatin caused dose-dependent declines in the mean inulin clearance values in the rat that were not attenuated with pharmacological doses of pentoxifylline. There were no differences in histology, urinary electrolyte excretion rates, and serum creatinine values between cisplatin toxicity groups treated with saline or pentoxifylline. Erythrocyte congestion was studied with 51Cr-labelled erythrocytes in rats given single doses of 5 mg/kg of cisplatin with and without pentoxifylline in an attempt to define the role of capillary sludging. The papilla, medulla, and cortex of kidneys of cisplatin-treated rats were markedly congested with 51Cr-erythrocytes; however, pentoxifylline treatment did not significantly reduce the congestion. These data suggest that although erythrocyte trapping is involved in cisplatin ARF, treatment with pharmacological doses of vascular decongestants does not fully attenuate the functional defect.


Permeability of cellulosic and non-cellulosic membranes to endotoxin subunits and cytokine production during in-vitro haemodialysis. The possibility of endotoxin transfer across haemodialysis membranes remains a controversial issue. Additional concern has arisen because of the recent introduction in clinical practice of highly permeable, synthetic dialysis membranes and of bacteria-contaminated bicarbonate concentrate with potential short-term and long-term hazards for haemodialysis (HD) patients. Therefore, we performed experiments in an in-vitro dialysis recirculation system using three different types of HD membranes, namely standard regenerated cellulose (Cuprophan, CU), polyacrylonitrile AN-69 (PAN), and polysulphone F-60 (PS). When radiolabelled lipopolysaccharide (125I M-LPS) from E. coli, together with 10 micrograms/ml unlabelled LPS, was added to the recirculating solution in the dialysis compartment, radioactivity could be detected in the blood compartment after 15 min and increased progressively with time up to respectively 6.7% (CU), 10.3% (PAN), and 10.3% (PS) of initial activity on the dialysate side. The addition of albumin to the solution on the blood side led to a decreased permeability of radioactivity (7.3% vs 10.3%), compared to the absence of albumin (tested only for PS membrane). Furthermore, 73% of 125I M-LPS transferred across the PS membrane in the presence of albumin was TCA-precipitable. In contrast, free iodine (Na 125I) incubated in an albumin-containing solution did not precipitate with albumin after the addition of TCA (precipitation of only 0.6%). Moreover, kinetics of transmembranous transfer of Na-125I were strikingly different from that of 125I M-LPS. Analysis by the method of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of the blood side solution, after LPS addition in the dialysis solution and 30 min of back-filtration, revealed the presence of several silver-stainable and autoradiographic bands of low-molecular-weight range, probably LPS fragments. Finally, the presence of LPS in the dialysate compartment led to a moderate increase in interleukin 1 (IL-1) and tumour necrosis factor alpha (TNF) concentrations in plasma as well as in monocyte culture supernatants after isolation from recirculating normal human whole blood exposed to CU, PAN, or PS membrane. In conclusion, our study provides evidence for the permeation of low-molecular-weight LPS subunits across cellulosic and non-cellulosic HD membranes. The clinical significance, if any, of such a transfer has, however, still to be demonstrated.


Blood flow, ultrafiltration and solute transport rate in continuous arteriovenous haemodiafiltration: the AN-69 flat-plate haemofilter. We measured blood flow, ultrafiltration rate and uraemic solute clearance at different dialysate flow rates during CAVHD using the AN-69 0.43 m2 flat plate haemofilter. As filter performance depends on clinical conditions and operational characteristics, data were analysed in terms of resistance to blood flow, membrane index of ultrafiltration, and diffusive mass transfer coefficients. An attempt was made to construct nomograms that may be used both to predict filter performance and to compare different haemofilters with each other.


Insulin-like growth factor I: a modulator of erythropoiesis in uraemic patients? Anaemia is a feature almost invariably complicating chronic renal failure. Its pathophysiology is multifactorial but the most important cause is erythropoietin (Epo) deficiency. However, either no relation or even a weakly positive relation generally exists between serum immunoreactive (i) Epo and haematocrit values in uraemic anaemia, whereas in anaemias of non-renal origin the correlation is most often strongly negative. Recent evidence indicates that growth hormone also stimulates erythropoiesis. Moreover, late erythroid progenitor cells (CFU-E) require insulin and/or insulin-like growth factor I (IGF-I) for development in vitro. IGF-I has been shown to have a synergistic action with Epo. We have measured serum iEpo and IGF-I levels in 17 haemodialysis patients with severe hyperparathyroidism (mean +/- SEM serum iPTH, 988 +/- 88 pg/ml). Mean age and duration of dialysis treatment were 46.1 +/- 3.4 and 8.8 +/- 1.0 years respectively. Mean haematocrit and haemoglobin values wer 28.1 +/- 1.7% and 9.39 +/- 0.54 g/dl respectively. Mean serum iEpo and IGF-I levels were 20.3 +/- 4.7 mU/ml and 320 +/- 20 ng/ml respectively (normal values for serum iEpo and IGF-I, 17.9 +/- 6 mU/ml and 91 +/- 23 ng/ml respectively). We found that serum IGF-I concentrations were well correlated with haematocrit values (r = 0.68, n = 15, P less than 0.004) whereas serum iEpo values were not (r = 0.41, n = 12, P = 0.18). IGF-I could therefore be an important factor regulating erythropoiesis in uraemic patients, at least when associated with severe hyperparathyroidism.


Effect of recombinant human erythropoietin therapy on ambulatory blood pressure and heart rate in chronic haemodialysis patients. Systemic hypertension as assessed by causal blood pressure measurements is a frequently reported side-effect of recombinant human erythropoietin (rHuEpo) treatment. We investigated the effect of rHuEpo treatment on the 24-h ambulatory blood pressure and heart rate profiles of 13 chronic haemodialysis patients. After 3-4 months of rHuEpo therapy it was found that the mean haematocrit had increased from 24.5 +/- 1.0% to 32.0 +/- 1.1% (P less than 0.005), while body-weight and control of uraemia as assessed by routine laboratory data remained unchanged. Despite gradual and incomplete correction of anaemia by use of low doses of rHuEpo, increases in the ambulatory systolic and diastolic blood pressure were found. The greatest increases affected day-time systolic blood pressure and night-time diastolic blood pressure, and these increases were significant (P less than 0.05). As a result, pulse pressure increased during day-time (P less than 0.05) while the night-time decline in diastolic blood pressure disappeared. An increase in peripheral resistance after partial correction of renal anaemia might explain these observations. rHuEpo therapy increased the percentage of abnormal ambulatory blood pressure measurements (defined as systolic blood pressure greater than 140 mmHg and/or diastolic blood pressure greater than 90 mmHg) from 33% to 52% (P less than 0.05) while in contrast, mean casual prehaemodialytic and posthaemodialytic blood pressure values remained unchanged. We conclude that changes in 24-h blood pressure profiles should be carefully assessed by ambulatory blood pressure monitoring in haemodialysis patients treated with rHuEpo, since these changes are likely to be missed when only causal blood pressures are measured.


Aqueous solute concentrations and evaluation of mass transport coefficients in peritoneal dialysis. The quantitative description of diffusive and convective mass transport of small solutes in peritoneal dialysis is dependent on accurate determination and appropriate expression of the concentration of investigated substances. For small solutes which easily equilibrate between dialysate and plasma the solute concentration in plasma should be expressed per volume of plasma water (aqueous concentration) and not per volume of whole plasma. Furthermore, the Donnan effect should be taken into account for electrolytes if measured by flame photometry. The common practice of expressing solute concentration per volume of whole plasma (plasma concentration) may result in substantial errors in calculated values of peritoneal transport parameters. To quantify these errors we compared plasma versus aqueous dialysate to plasma ratios (D/P), diffusive mass transport coefficients (KBD), and sieving coefficients (S) for 28 6-h single-dwell studies using glucose 3.86% dialysis fluid. For all substances except glucose non-corrected plasma D/P overestimated corrected aqueous D/P at 360 min by 2% (potassium and sodium) to 8% (creatinine) and, as assessed by the Pyle-Popovich model, non-corrected KBD overestimated true KBD by 12% (potassium) to 41% (urea). Similar results were also obtained for KBD estimation using the Garred model and KBD estimated during dialysate isovolaemia. The use of aqueous instead of plasma concentrations resulted in a substantial change of S for urea but not for the other investigated solutes. These results emphasise the importance of expressing small solute concentrations per volume of plasma water and not per volume of whole plasma in calculations of D/P ratios and mass transport coefficients.


Transfer of autologous haemoglobin from the peritoneal cavity during peritoneal dialysis. Transfer of autologous haemoglobin from the peritoneal cavity was evaluated retrospectively in 14 patients who received this marker intraperitoneally (group 1) during routine continuous ambulatory peritoneal dialysis (CAPD). Five additional patients were studied during acute peritonitis (group 2). A model for balance of both dialysate volume and amount of haemoglobin is developed to assess movement of the compound into lymph or adjacent tissues. Under the conditions of the study the transfer was slow (8 +/- 10 ml/h) in patients without peritonitis (group 1), and significantly faster (25 +/- 22 ml/h) in those with peritonitis (group 2). These clearance values are the upper limits for lymph flow.


Dialysate calcium reduction in CAPD patients treated with calcium carbonate and alfacalcidol. The use of oral calcium carbonate as a phosphate binder is often complicated by hypercalcaemia, particularly with concomitant use of vitamin D analogues. We previously found that stepwise reduction of dialysate calcium effectively countered this complication in haemodialysis patients, and have now assessed the strategy in CAPD patients. Seventeen patients underwent conversion from aluminium hydroxide to calcium carbonate and were followed for 5 months, with subsequent addition of alfacalcidol for a further 5 months. Standard CAPD dialysate (1.75 mM calcium) was used, reducing to 1.45 mM and, if necessary, to 1.00 mM in patients who became hypercalcaemic. While receiving calcium carbonate alone, 12 of the 17 patients became hypercalcaemic, this responding in four to dialysate calcium reduction to 1.45 mM. In the remaining eight patients, further reduction to 1.00 mM was required and in two patients even this failed to control hypercalcaemia adequately, necessitating reversion to aluminium hydroxide. Phosphate control remained unchanged, as did calcium x phosphorus product. There were transient increases of blood ionised calcium, and decreases of parathyroid hormone, with progressive reduction of serum aluminium and alkaline phosphatase. The addition of alfacalcidol (0.25 microgram/day) led to hypercalcaemia in six subjects, successfully countered by dialysate calcium reduction in four. The results show that standard CAPD dialysate calcium at 1.75 mM is too high for the majority of calcium carbonate treated patients and that substantial reductions of the dialysate calcium concentration are required if calcium carbonate is to be used effectively.


Effect of felodipine on renal haemodynamics and tubular sodium handling in cyclosporin-treated renal transplant recipients. The effect of a single oral dose of 10 mg of the calcium antagonist felodipine or placebo was investigated in 10 cyclosporin-treated renal transplant recipients before, during, and after an acute intravenous infusion of cyclosporin in a randomised, single-blind cross-over study. Renal plasma flow (RPF), glomerular filtration rate (GFR), renal tubular sodium and water handling as judged by the lithium clearance technique, and plasma concentrations of angiotensin II (AngII), aldosterone (Aldo), atrial natriuretic factor (ANF), and arginine vasopressin (AVP) were measured. Both RPF and GFR increased after felodipine (mean increase: RPF, 38.7%; GFR, 16.2%; P less than 0.01 for both) in spite of a significant decrease in both systolic and diastolic blood pressure (mean decrease 10.6% and 16.0% respectively, P less than 0.02 for both). Estimated by the lithium clearance technique felodipine induced a decrease in fractional reabsorption in the proximal tubules (mean 72.0% vs 63.0%, P less than 0.01), an increase in proximal output of fluid (mean 11.0 ml/min vs 16.0 ml/min, P less than 0.01), and a decrease in distal fractional reabsorption of sodium (mean 90.5% vs 83.9%, P less than 0.05) resulting in a significant natriuresis and diuresis. Ang II, Aldo, ANF, or AVP did not change. Intravenous infusion of cyclosporin per se did not influence any of the parameters. It is concluded that a single dose of felodipine in cyclosporin treated renal transplant recipients has beneficial effects on blood pressure, renal haemodynamics, and renal tubular sodium and water handling, which seems to compensate for some of the adverse effects of cyclosporin. It is suggested that these effects result from a direct vasodilatation and an effect on proximal tubular function.


Frusemide therapy and intact parathyroid hormone plasma concentrations in chronic renal insufficiency. It has been suggested that frusemide affects plasma parathyroid hormone (PTH) concentrations. To further investigate this issue we analysed plasma intact PTH in 77 patients with chronic renal failure (CCr 8.0-89.8 ml/min per 1.73 m2) as a function of frusemide therapy. The rate of increase of plasma PTH observed with progression of renal failure was faster in patients who received frusemide as compared to patients who did not receive the drug. The slope of the regression line of PTH on CCr was steeper (P less than 0.02) for patients with frusemide (n = 40, slope -0.34) than without frusemide (n = 37, slope -0.20). This effect was specific for frusemide therapy since therapy with other antihypertensive drugs (including thiazides and beta-blockers) was not correlated with PTH plasma concentrations. Frusemide therapy was also associated with a significantly greater urinary calcium excretion in uraemic patients but did not influence other parameters of calcium metabolism. To clarify mechanisms involved in the effect of frusemide on plasma PTH values, seven normal subjects were studied for 24 h before and for 24 h after oral administration of 80 mg frusemide. The main findings were: (1) Median PTH values were higher than on a control day (P less than 0.05) 3 h after frusemide (3.9 pmol/l vs 1.8) and 6 h after frusemide (4.0 vs 2.6); (2) ionised plasma calcium did not change significantly, whereas mean calcium/creatinine ratio increased from 0.20 to 0.46 after frusemide treatment through an increase in absolute calcium excretion; (3) plasma 1 alpha,25-dihydroxyvitamin D3, catecholamines, and magnesium concentrations did not change significantly after frusemide.(ABSTRACT TRUNCATED AT 250 WORDS)


Okadaic acid inhibits the release of TSH in response to TRH and K+ from rat anterior pituitaries. The effects of okadaic acid, a non-phorbol-12-tetradecanoate-13-acetate (non-TPA)-type tumor promoter and a potent inhibitor of protein phosphatases, on thyroid-stimulating hormone (TSH) secretion from the rat anterior pituitary were examined. Preincubation of anterior pituitaries with okadaic acid caused a time- and concentration-related decrease in a subsequent thyrotropin-releasing hormone (TRH)-stimulated TSH secretion, whereas it did not cause any changes in basal secretion of TSH. In addition, okadaic acid inhibited a subsequent high K(+)-induced TSH secretion. In contrast, ionomycin-induced TSH secretion was not inhibited by pretreatment with okadaic acid. The present results suggest that okadaic acid may block the release of TSH by inhibition of Ca2+ influx through voltage-sensitive and/or receptor-operated Ca2+ channels.


Pregnancy alters the density of opioid binding sites in the supraoptic nucleus and posterior pituitary gland of rats. Opioid receptor binding was measured in cryostat sections of supraoptic nucleus (SON) and posterior pituitary of virgin and pregnant rats by quantitative receptor autoradiography after in vitro incubation with [3H]etorphine or [3H](-)-bremazocine in the presence of unlabelled sub-type-selective agonists. Mu-selective [3H]etorphine-binding in the SON was reduced on the last day (21) of pregnancy vs. virgin controls (9.9 +/- 2.2 vs. 31.7 +/- 6.5 fmol/mg). Kappa-selective [3H](-)-bremazocine binding to the SON was not altered by pregnancy. Kappa-selective [3H](-)-bremazocine-binding to the posterior pituitary was less on day 16 of pregnancy vs. virgin females (19.1 +2- 5.2 vs. 74.4 +/- 16.2 fmol/mg). The results suggest mechanisms for the changes in actions of opioids on oxytocin neurones in pregnancy.


Sources and targets of nitric oxide in rat cerebellum. Nitric oxide (NO) mediates cell-cell signalling in the brain and stimulates cyclic GMP (cGMP) production in target cells. We have used NADPH-diaphorase (reduced nicotinamide adenine dinucleotide phosphate-diaphorase) histochemistry to identify NO-producing neurones and cGMP immunohistochemistry to locate the targets of NO in rat cerebellum. NADPH-diaphorase staining was prominent in granule cells and in the molecular layer. cGMP immunostaining in cerebellar slices stimulated with the NO donors, nitroprusside and SIN-1, was found in granule cells, glomeruli, fibres, Bergmann glia and in other astrocytes. The results provide visible evidence that NO mediates neuron-neuron and neuron-glia communication.


Investigation of beta-adrenergic modulation of synaptic transmission and postsynaptic induction of associative LTP in layer V neurones in slices of rat sensorimotor cortex. Long-term potentiation (LTP) of synaptic transmission is considered to be a neuronal model of learning. Recently, the probability of induction of associative LTP in layer V cells in sensorimotor neocortex was shown to be much higher in the awake cat than in the slice preparation. We hypothesised that the loss of extrinsic noradrenergic activity in the slice might account for this difference, particularly since a beta-adrenergic enhancement of field potentials has been seen in this preparation. We therefore bath-applied noradrenaline (NA) or the beta 1-adrenergic agonist, isoprenaline (ISO) to elucidate the cellular basis of the enhancement of field potentials, and to see if the drugs increased the probability of induction of associative LTP in slices. We found that NA and ISO produced a dose-dependent, reversible reduction of spike accommodation and an increase in excitability but had no effect on the depolarizing slope or peak amplitude of sub-threshold EPSPs, and that drug application did not increase the probability of induction of LTP. We conclude that: (1) the enhancement of field potentials and late components of EPSPs (7) can be explained by the known actions of beta-adrenergic drugs on membrane currents in layer V cells, and (2) the lower probability of induction of associative LTP in slices cf. the awake cat cannot be due solely to the loss of noradrenergic activity.


Inhibitory effect of insulin and cytoplasmic factor(s) on brain (Na(+) + K+) ATPase. (Na+ + K+)ATPase activity in cerebral cortex was modulated by insulin action depending on the Mg2+ concentration. Thus, in homogenates in the presence of 1-3 mM Mg2+, insulin stimulated the enzyme, whereas in the presence of 4-6 mM Mg2+ inhibition was observed. Exposure of synaptosomal membranes to the soluble fraction resulted in inhibition of ATPase activity in a dose-dependent manner. The inhibitory effect of insulin was regulated by a cytoplasmic factor in a dose-dependent manner. Similar variations to those obtained with a crude synaptosomal fraction were obtained by using a partially purified ATPase. These results indicated the importance of soluble factors in the modulation of ATPase by insulin and add more evidence in support for a role of insulin as a neuromodulator.


Resetting of the rat circadian clock after a shift in the light/dark cycle depends on the photoperiod. Adjustment of the circadian clock to shifts in the light/dark (LD) cycle was assessed from the rat pineal N-acetyltransferase (NAT) rhythm which is controlled by a pacemaker in the suprachiasmatic nucleus of the hypothalamus. Re-entrainment to an 8-h delay in the LD cycle took more than 3 days in rats maintained under a regime with 18 h of light and 6 h of darkness per day (LD 18:6) whereas it was completed within 3 days in those maintained under LD 12:12. Re-entrainment to an advance in the LD cycle proceeded through a transient diminution or almost disappearance of the NAT rhythm amplitude following a 5-h, 3-h and even a mere 2-h advance shift under LD 18:6, whereas no such diminution occurred under LD 12:12 even after a 5-h advance shift. Altogether, the data indicate that resetting of the circadian clock after shifts in the LD cycle depends on the photoperiod.


Immunocytochemical studies on light-induced changes in phosphatidylinositol 4,5-bisphosphate immunoreactivity in the visual system of normal and norpA mutant of Drosophila. The distribution of phosphatidylinositol 4,5-bisphosphate (PIP2) in the visual system of Drosophila was studied by indirect immunofluorescence staining using a monoclonal antibody against PIP2. The retina of the compound eye and the cortical regions of the optic lobe were heavily stained by the antibody. In the retina, photoreceptor cells were stained with the antibody, but non-neuronal cells such as pigment cells and cone cells were not stained. The staining intensity in the light-adapted photoreceptor cells was lower than that in dark-adapted cells in normal flies, whereas no such difference in immunoreactivity was observed in the norpAEE5 mutant, whose photoreceptor cells are deficient in phospholipase C. These results suggest that the PIP2 in the photoreceptor cells is hydrolyzed by phospholipase C coded by the norpA gene upon light stimulation.


Conservative surgery plus adjuvant therapy for vulvovaginal rhabdomyosarcoma, diethylstilbestrol clear cell adenocarcinoma of the vagina, and unilateral germ cell tumors of the ovary. Significant progress has been made in the 1980s in early-stage vulvovaginal rhabdomyosarcoma, diethylstilbestrol (DES) clear cell adenocarcinoma of the vagina, and unilateral germ cell tumors of the ovary. In an early state of vulvovaginal rhabdomyosarcoma, systemic vincristine, dactinomycin, and cyclophosphamide (VAC) chemotherapy followed by local excision or local radiation results in a high cure rate with retention of future fertility. Similarly, early-stage DES-related adenocarcinoma of the vagina treated by wide local excision and localized vaginal radiation also results in retention of fertility and a high cure rate. Finally, significant progress has been made in unilateral germ cell tumors of the ovary in which surgical treatment by unilateral salpingo-oophorectomy followed by cisplatin, etoposide, and bleomycin results in not only high cure rates and retention of fertility but will probably be standard therapy for all germ cell tumors of the ovary, including dysgerminoma, a disease most frequently treated in the past by radiation therapy with loss of subsequent fertility.


Effect of dietary fiber on the disposition and excretion of a food carcinogen (2-14C-labeled MeIQx) in rats. We studied to what extent dietary fiber may affect uptake, retention, and excretion of a food carcinogen (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, MeIQx) occurring in fried meat. Four diets--one fiber-free control and three containing either insoluble dietary fiber isolated from sorghum (100 g/kg) and wheat bran (100 g/kg) or the highly soluble pectin (50 g/kg)--were investigated. The fiber diets were given in amounts of 10 g/day to rats. Thus, each rat received 1 or 0.5 g fiber and 100 micrograms 2-14C-labeled MeIQx uniformly mixed in its daily diet. A 4-day adaptation period with unlabeled MeIQx was followed by a 5-day experimental period with 14C-labeled MeIQx, during which urine and feces were collected separately for analysis of radioactivity and mutagenicity. Furthermore the composition and the fermentability of the dietary fiber were determined. The present study shows that a diet containing fiber, especially fiber isolated from sorghum and wheat bran, affects the excretion pattern of the food carcinogen MeIQx in a manner suggesting a lower uptake and a decreased transit time through the gastrointestinal tract in a more diluted form than a nonfiber diet. Furthermore, less radioactivity was retained in the kidneys with sorghum and wheat bran than with the other two diets. On the other hand, none of these types of dietary fiber affected the retention of the hepatocarcinogen MeIQx in the liver 24 hours after the last oral intake. DNA adducts were formed to a higher extent in the kidney than in the liver. The highest levels were found in animals given the wheat bran diet.


[Chronic heart failure: biological bases of myocardial function]. This review paper deals with recent data concerning the physiopathology of chronic heart failure. Broadly speaking, the cause of chronic heart failure is arterial hypertension and/or coronary disease. Myocardial hypertrophy is only one example of biological adjustment to the environment, and chronic heart failure, an adaptation disease, indicates its limits. Diastolic dysfunction includes 3 elements. Relaxation is slowed down by a decrease in density of Ca(2+)-ATPase in the sarcoplasmic reticulum and by a fall in Na+/Ca2+ exchange activity. The decline of tissue compliance is mainly explained by probably hormonal changes in collagen. Changes in isomyosins account for abnormalities of atrial contraction. At the beginning of mechanical overload, the fall in systolic function enables the heart to produce a normal tension. The determinant element is slowing down of intracellular calcium movements. Enlarged hearts generate arrhythmias. The origin is probably the presence of unstable calcium homeostasis.


[Procollagen III-peptide and angiotensin converting enzyme in bronchoalveolar lavage in sarcoidosis]. In 39 cases with active and 36 cases with inactive sarcoidosis the angiotensin converting enzyme (ACE) and in 56 of them (21 active/35 inactive sarcoidosis) procollagen-III-peptide (P III P) content as well as the total protein content of the broncho-alveolar lavage (BAL) and of the serum were determined. These results were compared with those of a control group of 53 patients (ACE group) resp. 25 patients (P III P-group). Both the relative ACE content (ACE related to 1 g of protein) and the relative P III P content (P III P related to 1 mg of protein) showed significant increase in cases of acute sarcoidosis compared to the other two groups of patients. However, none of the three groups showed statistically significant differences in the other parameters. ACE in the BAL can therefore be regarded as a good indication of the activity of sarcoidosis. This is also true of P III P in the BAL as an indication of fibrogenesis.


[Transesophageal echography in staging of bronchial cancers]. The kind of relation of central lung cancer (c) to the walls of the central pulmonary arteries (PA) and the aorta is an important information prior to operative or interventional (laser/afterloading) therapy. As computed tomography (CT) and angiography are often inaccurate in the assessment of PA-infiltration, we assessed the diagnostic value of transesophageal echography (TEE) in the staging of LC. 16 patients (pts.) were investigated using TEE in addition to CT or magnetic resonance imaging (MRI). Eleven pts. had central LC, 3 peripheral LC, 1 anterior mediastinal mass and 1 central pneumonia (cancer excluded). 2 pts. with central LC were unable to swallow the probe. In 9/9 pts. with central LC, 1/3 pts. with peripheral LC and 1 pt. with enlarged anterior mediastinum the tumour mass could be visualized. In the pt. with a centrally located infiltrate on chest radiogram TEE demonstrated enlarged hilar lymph nodes, but excluded a central tumour. Main PA branches could be identified in all 14/14 pts. Central left or right PA were compressed slightly in 3 pts. and severely in 2 pts., with a near total occlusion in one (confirmed by MRI/CT). TEE revealed PA-infiltration in 2 pts. and aortic wall infiltration in 2 other pts. Despite adjacent tumour mass aortic wall infiltration was excluded in 2 pts. Enlarged hilar lymph nodes could be demonstrated in 2/9 pts. with central LC, whereas CT/MRI showed enlarged mediastinal lymph nodes in 7/9 pts. In conclusion, TEE is able to visualize central lung cancer and gives useful additional informations about the kind of relation to central PA and the aorta.(ABSTRACT TRUNCATED AT 250 WORDS)


SNC1, a yeast homolog of the synaptic vesicle-associated membrane protein/synaptobrevin gene family: genetic interactions with the RAS and CAP genes. SNC1, a gene from the yeast Saccharomyces cerevisiae, encodes a homolog of vertebrate synaptic vesicle-associated membrane proteins (VAMPs) or synaptobrevins. SNC1 was isolated by its ability to suppress the loss of CAP function in S. cerevisiae strains possessing an activated allele of RAS2. CAP is a component of the RAS-responsive S. cerevisiae adenylyl cyclase complex. The N-terminal domain of CAP is required for full cellular responsiveness to activated RAS proteins. The C-terminal domain of CAP is required for normal cellular morphology and responsiveness to nutrient extremes. Multicopy plasmids expressing SNC1 suppress only the loss of the C-terminal functions of CAP and only in the presence of activated RAS2.


Urate-null rosy mutants of Drosophila melanogaster are hypersensitive to oxygen stress. It has been proposed that uric acid is an important scavenger of deleterious oxygen radicals in biological systems [Ames, B. N., Cathcart, R., Schwiers, E. & Hochstein, P. (1981) Proc. Natl. Acad. Sci. USA 78, 6858-6852]. We report here an in vivo investigation of the oxygen defense role of uric acid through an analysis of mutants of the rosy (ry) gene of Drosophila melanogaster. The ry gene is the structural gene for the molybdoenzyme, xanthine dehydrogenase; xanthine dehydrogenase-null ry mutants are therefore unable to synthesize urate. The rationale of our approach was to measure the response of urate-null ry mutants to extraordinary oxygen stress as imposed by exposure to radical-generating agents and as conferred by a genetic defect in superoxide dismutase, an established oxygen defense function. We show that urate-null mutants of the ry locus are hypersensitive to paraquat, ionizing radiation, and hyperoxia. Furthermore, compound mutants doubly deficient for uric acid and Cu/Zn-containing superoxide dismutase are synthetic lethals, which are unable to complete metamorphosis under normal growth conditions. These experiments demonstrate unambiguously the importance of urate in oxygen defense in vivo and support our earlier proposal that the molybdoenzyme genetic system plays a critical role in oxygen defense in Drosophila. They also form the basis for our proposal that metamorphosis in Drosophila imposes a crisis of oxygen stress on the developing imago against which uric acid plays an important organ-specific defense. Finally, the results provide a basis for understanding the syndrome of phenotypes, including the hallmark dull brown eye color, which characterizes mutants of this classic genetic system of Drosophila.


Immortal rat hippocampal cell lines exhibit neuronal and glial lineages and neurotrophin gene expression. Clonal cell lines of rat embryonic hippocampal origin have been developed by using retroviral transduction of temperature-sensitive simian virus 40 large tumor antigens. The cell lines undergo morphological differentiation at the nonpermissive temperature and in response to differentiating agents. Immunocytochemical analysis indicates that various lines are derived from progenitors of neuronal, glial, and bipotential lineages. Selected neuronal lines differentiate in response to diffusible factors released by primary glia, and one line of glial lineage supports the maturation of primary neurons in culture. Selected cell lines exhibit different patterns of neurotrophin gene expression that change after differentiation. In some lines, the relative levels of neurotrophin 3 and brain-derived neurotrophic factor message expression may reflect the developmental or regional differential expression seen for these genes in the hippocampus in situ. These hippocampal cell lines, which express markers indicative of commitment to neuronal or glial lineages, are valuable for studies of development and plasticity in these lineages, as well as for studies of the regulation of neural trophic interactions.


Efficiency of binding the retinoblastoma protein correlates with the transforming capacity of the E7 oncoproteins of the human papillomaviruses. The human papillomaviruses (HPVs) associated with genital tract lesions can be classified as either "high risk" or "low risk" based on their association with human anogenital cancer. The E7 proteins of the high-risk and the low-risk viruses are quite similar in their amino acid composition and structural organization yet differ in their transforming potential and in a number of biochemical properties. A series of chimeric proteins consisting of segments of the high-risk HPV-16 and the low-risk HPV-6 E7 proteins were constructed in order to define which domains within the amino-terminal half of E7 were responsible for the different biological and biochemical properties. The E7 oncogenic capacity, which was determined by assaying transformation of baby rat kidney cells in cooperation with an activated ras oncogene, segregated with the retinoblastoma tumor suppressor protein (pRB) binding domain of the HPV-16 E7 protein. A comparison of the pRB binding sites of the sequenced genital tract HPVs revealed a consistent amino acid difference (aspartic acid/glycine) between the high-risk and low-risk viruses. Single amino acid substitution mutations were generated at this position in the HPV-6 and HPV-16 E7 proteins, and this single amino acid residue was shown to be the principal determinant responsible for the differences in the apparent pRB binding affinity and transformation capacity distinguishing the HPV E7 proteins of the high-risk and low-risk HPVs.


Natural human antibodies to gamma interferon interfere with the immunomodulating activity of the lymphokine. Natural antibodies to gamma interferon (IFN-gamma) were found in patients suffering from different viral diseases and, at a lower titer, in healthy individuals. Such antibodies were affinity-purified and studied for their capability to interfere in vitro with the antiviral and immunomodulating activity of IFN-gamma. Data obtained show that these human anti-IFN-gamma antibodies have no inhibitory effect on the antiviral activity of IFN-gamma. On the contrary, they are able to inhibit the expression of Fc receptor sites and HLA-DR antigens induced by IFN-gamma on the U-937 cells, a human monocytoid/macrophage-derived cell line. These antibodies can also interfere in a mixed lymphocyte culture (MLC) with the proliferation of lymphocytes and the generation of cytotoxic lymphocytes. However, they showed only a moderate inhibitory effect on the cytotoxicity generated in MLC to K-562 cells. Human antibodies capable of interfering with the immunomodulating activities of IFN-gamma might open up a new field in clinical therapy for those diseases that carry evidence of activated cell-mediated immunity.


Germ-line mutations of the APC gene in 53 familial adenomatous polyposis patients. We searched for germ-line mutations of the APC gene in 79 unrelated patients with familial adenomatous polyposis using a ribonuclease protection analysis coupled with polymerase chain reaction amplifications of genomic DNA. Mutations were found in 53 patients (67%); 28 of the mutations were small deletions and 2 were 1- to 2-base-pair insertions; 19 were point mutations resulting in stop codons and only 4 were missense point mutations. Thus, 92% of the mutations were predicted to result in truncations of the APC protein. More than two-thirds (68%) of the mutations were clustered in the 5' half of the last exon, and nearly two-fifths of the total mutations occurred at one of five positions. This information has significant implications for understanding the role of APC mutation in inherited forms of colorectal neoplasia and for designing effective methods for genetic counseling and presymptomatic diagnosis.


Molecular diversity of L-type Ca2+ channel transcripts in human fibroblasts. The nucleotide sequence of cDNA encoding the human fibroblast Ca2+ channel of L type (HFCC) has been determined. It is highly homologous to L-type channels previously cloned from rabbit lung and heart as well as from rat brain. At least four sites of molecular diversity were identified in the nucleotide sequence of HFCC. Three of these include regions encoding the transmembrane segments IIS6, IIIS2, and IVS3, which are known to be important for channel gating properties. The positions of these sites correlate with RNA splice sites, indicating that the molecular diversity of the transcripts is a result of alternative splicing. The fourth diversity region is located at the C-terminal region and comprises insertions and deletions. It is suggested that these variations may give rise to multiple subforms of HFCC with altered electrophysiological properties.


Adenovirus E1A, simian virus 40 tumor antigen, and human papillomavirus E7 protein share the capacity to disrupt the interaction between transcription factor E2F and the retinoblastoma gene product. The adenovirus E1A gene product, the simian virus 40 large tumor antigen, and the human papillomavirus E7 protein share a short amino acid sequence that constitutes a domain required for the transforming activity of these proteins. These sequences are also required for these proteins to bind to the retinoblastoma gene product (pRb). Recent experiments have shown that E1A can dissociate complexes containing the transcription factor E2F bound to pRb, dependent on this conserved sequence element. We now show that the E7 protein and the simian virus 40 large tumor antigen can dissociate the E2F-pRb complex, dependent on this conserved sequence element. We also find that the E2F-pRb complex is absent in various human cervical carcinoma cell lines that either express the E7 protein or harbor an RB1 mutation, suggesting that the loss of the E2F-pRb interaction may be an important aspect in human cervical carcinogenesis. We suggest that the ability of E1A, the simian virus 40 large tumor antigen, and E7 to dissociate the E2F-pRb complex may be a common activity of these viral proteins that has evolved to stimulate quiescent cells into a proliferating state so that viral replication can proceed efficiently. In circumstances in which a lytic infection does not proceed, the consequence of this action may be to initiate the oncogenic process in a manner analogous to the mutation of the RB1 gene.


Excision of Tn10 from the donor site during transposition occurs by flush double-strand cleavages at the transposon termini. Tn10 transposition is accomplished without extensive replication of the transposon sequences. Replicative cointegrate formation is precluded by efficient separation of transposon sequences from flanking donor DNA at an early stage in the transposition reaction. We report here that excision of Tn10 from its donor site occurs by a pair of flush double-strand breaks. Breaks occur at each end of the element precisely between the terminal base pair of the element and the first base pair of flanking DNA. This observation provides definitive evidence that cleavage of both strands of the element occurs under the direct control of Tn10 transposase protein. It is highly likely that transposase itself is directly responsible for these cleavages. The implications of this possibility are discussed.


Fatty acids activate a chimera of the clofibric acid-activated receptor and the glucocorticoid receptor. Peroxisome proliferators such as clofibric acid, nafenopin, and WY-14,643 have been shown to activate PPAR (peroxisome proliferator-activated receptor), a member of the steroid nuclear receptor superfamily. We have cloned the cDNA from the rat that is homologous to that from the mouse [Issemann, I. & Green, S. (1990) Nature (London) 347, 645-650], which encodes a 97% similar protein with a particularly well-conserved putative ligand-binding domain. To search for physiologically occurring activators, we established a transcriptional transactivation assay by stably expressing in CHO cells a chimera of rat PPAR and the human glucocorticoid receptor that activates expression of the placental alkaline phosphatase reporter gene under the control of the mouse mammary tumor virus promoter. Testing of compounds related to lipid metabolism or peroxisomal proliferation revealed that 150 microM concentrations of arachidonic or linoleic acid but not of dehydroepiandrosterone, cholesterol, or 25-hydroxy-cholesterol, activate the receptor chimera. In addition, saturated fatty acids induce the reporter gene. Shortening the chain length to n = 6 or introduction of an omega-terminal carboxylic group abolished the activation potential of the fatty acid. In conclusion, the present results indicate that fatty acids can regulate gene expression mediated by a member of the steroid nuclear receptor superfamily.


Estrogen receptors colocalize with low-affinity nerve growth factor receptors in cholinergic neurons of the basal forebrain. The rodent and primate basal forebrain is a target of a family of endogenous peptide signaling molecules, the neurotrophins--nerve growth factor, brain-derived neurotrophic factor, and neurotrophin 3--and of the gonadal steroid hormone estrogen, both of which have been implicated in cholinergic function. To investigate whether or not these ligands may act on the same neurons in the developing and adult rodent basal forebrain, we combined autoradiography with 125I-labeled estrogen and either nonisotopic in situ hybridization histochemistry or immunohistochemistry. We now report colocalization of intranuclear estrogen binding sites with the mRNA and immunoreactive protein for the low-affinity nerve growth factor receptor, which binds all three neurotrophins, and for the cholinergic marker enzyme choline acetyltransferase (acetyl-CoA:choline O-acetyltransferase, EC 2.3.1.6). Colocalization of estrogen and low-affinity nerve growth factor receptors implies that their ligands may act on the same neuron, perhaps synergistically, to regulate the expression of specific genes or gene networks that may influence neuronal survival, differentiation, regeneration, and plasticity. That cholinergic neurons in brain regions subserving cognitive functions may be regulated not only by the neurotrophins but also by estrogen may have considerable relevance for the development and maintenance of neural substrates of cognition. If estrogen-neurotrophin interactions are important for survival of target neurons, then clinical conditions associated with estrogen deficiency could contribute to the atrophy or death of these neurons. These findings have implications for the subsequent decline in those differentiated neural functions associated with aging and Alzheimer disease.


In vitro replication of adeno-associated virus DNA. An in vitro assay for adeno-associated virus (AAV) DNA replication has been developed. The substrate is a plasmid containing the duplex form of AAV DNA in pBR322. The AAV insert is excised or rescued from the plasmid by extracts of uninfected cells. Replication was assayed by production of full-length excised AAV DNA resistant to Dpn I digestion. The following results were obtained. (i) Only extracts of cells coinfected with AAV and adenovirus replicated the excised insert. (ii) Density label experiments showed semiconservative replication. (iii) Only the excised AAV insert was replicated; pBR322 sequences were not. (iv) Replication was dependent on the presence of the AAV terminal repeat. (v) If the terminal 55 bases were deleted from both ends of the AAV insert, no rescue took place: replication occurred and both AAV and pBR322 sequences were replicated. We conclude that the AAV terminal repeat is essential for DNA replication but that under some conditions an initiation mechanism that does not involve hairpin priming may be used.


Finding errors in DNA sequences.

Finding errors in DNA sequences. An algorithm is described that can detect certain errors within coding regions of DNA sequences. The algorithm is based on the idea that an insertion or deletion error within a coding sequence would interrupt the reading frame and cause the correct translation of a DNA sequence to require one or more frameshifts. If the coding sequence shows similarity to a known protein sequence then such errors can be detected by comparing the conceptual translations of DNA sequences in all six reading frames with every sequence in a protein sequence data base. We have incorporated these ideas into a computer program, called DETECT, that can serve as an aid to the experimentalist who is determining new DNA sequences so that obvious errors may be located and corrected. The program has been tested using raw experimental data and against sequences from the European Molecular Biology Laboratory data base, annotated as containing frameshifts. We have also tested it using unidentified open reading frames that flank known, annotated genes in the GenBank data base. Many potential errors are apparent and in some cases functions can be suggested for the "corrected" versions of these reading frames leading to the identification of new genes. As more sequences are determined the power of this method will increase substantially.


Assessment of a cholinergic contribution to chlordiazepoxide-induced deficits of place learning in the Morris water maze. This investigation sought to characterize the interaction between benzodiazepine and cholinergic systems in place learning in the Morris water maze. In the first experiment, rats were treated with scopolamine (1 mg/kg) alone or concomitantly with one of two doses of flumazenil (15 and 30 mg/kg) or with chlordiazepoxide (5 mg/kg) alone or concomitantly with flumazenil (15 mg/kg). Chlordiazepoxide and scopolamine severely impaired place learning but not cue learning. The low dose of flumazenil completely reversed the impairment produced by chlordiazepoxide and both high and low doses of flumazenil attenuated the place learning deficit produced by scopolamine. Neither dose of flumazenil affected place learning when administered alone. In the second experiment, rats were administered chlordiazepoxide (5 mg/kg) or scopolamine (1 mg/kg) alone or concomitantly with one of four doses of physostigmine (0.05, 0.10, 0.25, and 0.5 mg/kg). Once again, both chlordiazepoxide and scopolamine impaired place but not cue learning. Physostigmine reversed the impairment produced by scopolamine in a dose-dependent manner but failed at every dose to attenuate the impairment produced by chlordiazepoxide. The higher doses of physostigmine impaired place learning when administered alone. None of the drug treatments impaired cue learning. Together, these results suggest that the scopolamine-induced impairment of place learning is due to an increase in benzodiazepine/GABA activity, and contradict the notion that benzodiazepines impair memory by cholinergic mechanisms.


Autoradiographic mapping of neurotransmitter system receptors in mammalian brain. Regional localization of neurotransmitter system receptors was visualized in the gerbil grain and in the rat brain using receptor autoradiography. [3H]Quinuclidinyl benzilate (QNB), [3H]cyclohexyladenosine (CHA), [3H]muscimol, [3H]MK-801, [3H]SCH 23390, [3H]PN200-110, [3H]spiperone, and [3H]naloxone were label muscarinic receptors, adenosine A1 receptors, GABAA receptors, N-methyl-D-aspartate (NMDA) receptors, dopamine D1 receptors, L-type calcium channels, spirodecanone receptors, and opioid receptors, respectively. Regional localization of [3H]QNB, [3H]muscimol, [3H]MK-801, [3H]SCH 23390, and [3H]PN200-110 binding sites in the gerbil brain was relatively similar to that in the rat brain. In contrast, the autoradiographic distribution of [3H]spiperone and [3H]naloxone binding sites in the gerbil was quite different from that in the rat. This phenomenon was found especially in the hippocampus and the cerebellum. The results suggest that the gerbil differs from the rat with respect to spirodecanone and opioid binding sites in the hippocampus and the cerebellum. This finding may help to further elucidate the species differences and relationships for brain function and behavioral pharmacology.


Direct evidence for mediation of an anticonflict effect of baclofen by GABAb receptors. The present article reports an experiment on the effects of baclofen (0, 0.5, 1.0, and 2.0 mg/kg) on punished drinking in rats and the modification of these by delta-amino-n-valeric acid (DANVA) (0 and 10.0 mg/kg). Baclofen significantly enhanced punished drinking and this increase was abolished by DANVA, which had no intrinsic anxiogenic activity. It is concluded that GABAb receptors probably mediate this effect of baclofen and that such receptors may be a potential site of anxiolytic drug action.


Neoplastic seeding complicating percutaneous ethanol injection for treatment of hepatocellular carcinoma. The authors describe a case of subcutaneous neoplastic seeding in the abdominal wall in a 67-year-old man with posthepatitic liver cirrhosis complicated by a single nodule of well-differentiated hepatocellular carcinoma. He was treated with percutaneous ethanol injection (PEI) performed under ultrasound guidance. The neoplastic seeding developed along the needle track used to carry out fine-needle biopsy and PEI and was diagnosed 6 months after the beginning of treatment.


Hepatocellular carcinoma: correlation of MR imaging and histopathologic findings. Seventy-two histologically proved nodular hepatocellular carcinomas (HCCs) were studied with magnetic resonance (MR) imaging at 1.5 T. Capsules were present in 56 of the 72 tumors. Thirty-seven capsules were depicted on T1-weighted spin-echo MR images, and 16 were depicted on T2-weighted MR images. Visualization was dependent on thickness and structure of the capsules. Of the 72 tumors, 36 had a mosaic pattern. A mosaic pattern was visualized in 12 of the 36 tumors on T1-weighted images and in 27 of the 36 tumors on T2-weighted images. Six tumors were determined to be histologic grade 1, and all were hyperintense on T1-weighted images, regardless of whether intracellular fat deposits were present. Four of the six grade 1 tumors were isointense on T2-weighted images. In contrast, grades 2 and 3 tumors had various signal intensities on T1-weighted images and most were hyperintense on T2-weighted images. Twenty-one of 32 tumors (66%) with focal areas of increased signal intensity on T2-weighted images had intratumoral dilated sinusoids at histologic examination.


Immunocytochemical characterization of cytomegalovirus (CMV) infected giant cells in perinatal acquired human immunodeficiency virus (HIV) infection. In a pediatric case of necrotizing CMV myelitis after perinatal HIV infection characteristic cytomegalic cells, which could not be attached to a particular cell line by cell morphology, were studied after immunostaining with monoclonal and polyclonal antibodies raised against GFAP, S100 protein, NSE, synaptophysin, factor VIII, vimentin, macrophages, leukocytes, CMV, HSV I + II, toxoplasma, and HIV 1 gp41. Astrocytes, oligodendrocytes, neurons, ependymal and endothelial cells, macrophages, and Schwann cells stained positively with CMV antiserum. With regard to their immunological features the majority of cytomegalic cells ("owl eye cells") was identified as astrocytes, and in decreasing frequency, the remainder was characterized as macrophages, mesenchymal, and endothelial cells. It is concluded that CMV giant cells represent one phase of virus induced cell transformation, not only one single, but numerous cell types are exposed to after CMV infection.


Inhibition of prostaglandin delta 13 reductase activity in rabbit kidney cortex by glutathione disulfide. t-Butyl hydroperoxide and H2O2-Fe(2+)-EDTA-glutathione system which produces hydroxyl radicals did not affect the 15-hydroxy prostaglandin dehydrogenase activity in rabbit kidney cortex. On the other hand, H2O2-Fe(2+)-EDTA-glutathione system inhibited the prostaglandin delta 13 reductase activity. Mannitol, a scavenger of hydroxyl radicals, had no effect on the inhibitory action of this system, indicating that the effect of H2O2-Fe(2+)-EDTA-glutathione system on the prostaglandin delta 13 reductase may not be due to produced hydroxyl radicals. As a result of further investigation, it was shown that glutathione disulfide, which is synthesized concomitantly with hydroxyl radicals from H2O2-Fe(2+)-EDTA-glutathione, inhibited the prostaglandin delta 13 reductase activity. These results suggest that hydroperoxides and hydroxyl radicals may not be likely candidates for the modulator of the catabolism of prostaglandins in the kidney cortex, and that glutathione disulfide has the potential to modulate the prostaglandin catabolism by affecting the prostaglandin delta 13 reductase activity.


External calcium dependence of the uterine contraction induced by prostaglandins E2 and F2 alpha and its antagonism with natural progestins. Prostaglandins (PGs) E2 and F2 alpha are strong inducers of uterine contraction by promoting a Ca2+ increase into the cell through specific receptors coupled with the calcium channels. On the contrary, progesterone and 5 beta-reduced progestins promote smooth muscle relaxation by blocking the ion calcium influx. Thus, this study was designed to emphasize the importance of external calcium in the PGs-induced rat uterus contraction. Likewise, also studied was the antagonism and the interaction between PGs and progestins (progesterone and its 5 alpha and 5 beta-reduced derivatives) in the myometrium. Results showed that uterine contraction induced by PGs depends on external calcium, since verapamil or extracellular calcium depletion abolished the PGs effect. Regarding the PGs-progestins antagonism, it was observed that pregnanedione, pregnanolone and epipregnanolone were quite effective for counteracting of PGs-induced contraction. However, progesterone was effective in a middle range, whereas 5 alpha-reduced progestins (allopregnanedione and allopregnanolone) were almost ineffective. It has been concluded that the participation of PGs and progestins in the modulation of uterine contraction might be achieved through the control of calcium influx by opening (PGs) or blocking (progestins) receptor-operated calcium channels.


Maturation of functional antibody affinity in animals immunised with synthetic foot-and-mouth disease virus. A good correlation exists between specific neutralising antibody titre and protection against challenge with foot-and-mouth disease virus (FMDV) in infected or virus-vaccinated cattle, but not in the case of animals immunised with synthetic FMDV peptides. Therefore, mechanisms other than simple neutralisation are likely to be important in vivo. Antibody affinity may influence the protective capacity of sera from immunised animals and experiments were carried out to measure the functional affinity for synthetic FMDV peptide of sera from guinea pigs and cattle given various synthetic vaccines. In guinea pigs given a single dose of synthetic vaccine, antibody affinity increased with time after immunisation. In cattle, however, administration of a second dose of peptide 21 days after the first markedly retarded the process of affinity maturation. For guinea pig sera of equivalent neutralising activity, those of higher functional affinity had higher protective indices than those of lower functional affinity. Knowledge of the importance of antibody affinity in protection against FMD is important for an improved understanding of the mechanisms of protection and for the design of novel vaccines.


Air sampling procedure for evaluation of viral excretion level by vaccinated pigs infected with Aujeszky's disease (pseudorabies) virus. Five groups of eight fattening pigs were vaccinated and then infected with Aujeszky's disease virus. Viral excretion was evaluated by two means: deep nasal swabbing and air sampling. It appeared that infectious airborne virus could be recovered from day 1 to day 6 after infection in the isolated units where control animals were raised. In vaccinated animals, airborne particles were also detected but the amount and duration varied in relation to their immune status at the day of virulent challenge: viral excretion was significantly lower in pigs presenting a high antibody level (1/16 to 1/64) just before infection. Results obtained with nasal swabs and with air samples were closely related. Despite its low sensitivity, the air sampling procedure could be considered as an efficient tool for reflecting infectious viral pressure in a confined atmosphere.


Effects of dietary molybdenum on nematode and host during Trichostrongylus vitrinus infection in lambs. The addition of molybdenum (0.05 mmol kg-1 dry matter) to the diet of lambs exposed for four weeks to a trickle (2500 third stage larvae per day) infection with Trichostrongylus vitrinus reduced the number and length of adult worms retrieved from the small intestine 11 days later: both effects were particularly marked in female worms from female lambs (P less than 0.01). Worms from lambs given molybdenum contained less proteinase enzyme activity and secreted less proteinases in culture irrespective of the sex of the host. Pathogenicity was not attenuated by molybdenum. Damage to the intestinal mucosa was severe in both dietary groups but infected females given molybdenum developed lower plasma albumin concentrations and lighter dressed carcases than those not given molybdenum. Neither the effects on the parasite nor those on the host could be attributed simply to molybdenum-induced copper depletion, using conventional measures of copper status. Molybdenum may be toxic to T vitrinus but may also facilitate or enhance the inflammatory process limiting larval establishment or increasing parasite rejection.


Transcatheter arterial embolization in hepatocellular carcinoma: a long-term follow-up. Transcatheter arterial embolization (TAE) was performed in 1,058 cases of hepatocellular carcinoma (HCC), and the prognosis after TAE in HCC was then assessed from the long-term follow-up data. The results indicated that the prognosis was better for small tumors, and was significantly better for capsulated than noncapsulated tumors. For this reason, the prognosis of small tumors was not always better than that of large tumors. The prognosis worsened as the severity of portal obstruction increased. The prognosis improved with the use of Lipiodol (iodized oil). It was found that the use of anticancer drugs in combination with TAE, however, did not significantly affect the prognosis as compared with TAE alone. The 1-, 2-, 3-, 4-, and 5-year overall survival rates were 70%, 51%, 29%, 14%, and 5%, respectively.


Enhanced degradation of the ferritin repressor protein during induction of ferritin messenger RNA translation. Induction of ferritin synthesis in cultured cells by heme or iron is accompanied by degradation of the ferritin repressor protein (FRP). Intermediates in the degradative pathway apparently include FRP covalently linked in larger aggregates. The effect of iron on FRP degradation is enhanced by porphyrin precursors but is decreased by inhibitors of porphyrin synthesis, which implies that heme is an active agent. These results suggest that translational induction in this system may be caused by enhanced repressor degradation. While unique among translational regulatory systems, this process is common to a variety of other biosynthetic control mechanisms.


Giant cell carcinoma of the lung exhibiting a large, well-circumscribed photon deficiency in ventilation/perfusion scintigrams. Giant cell carcinoma is a specific clinicopathological entity because of its fulminating clinical behavior and peripheral location in the lung. We present two patients with rapidly fatal giant cell carcinoma; each 99mTcMAA and/or 133Xe ventilation scintigrams showed a large, discrete, photon-deficient area in the lung. This unique scintigraphic finding may result from the peripheral location and rapid grawth of the giant cell tumor. Since giant cell carcinoma is a specific clinicopathological entity and of definite significance in establishing the prognosis of patients with lung cancer, 99mTcMAA and/or 133Xe scintigraphic findings may be useful in revealing such a tumor.


Cholangiocarcinoma developing after simple excision of a type II choledochal cyst. We have reported the fourth case of cholangiocarcinoma associated with a resected diverticulum of the common bile duct (type II choledochal cyst). The malignancy developed 2 years after adenomatous hyperplasia was demonstrated by simple excision of the cyst. We hypothesize that adenomatous hyperplasia is an early phase of malignant transformation in such patients. We believe the finding of adenomatous hyperplasia is of considerable importance in determining the extent of surgical resection.


Association of neurotensin receptors with VIP-containing neurons and serotonin-containing axons in the suprachiasmatic nucleus of the rat. The aim of the present study was to identify cellular elements bearing high-affinity neurotensin (NT) binding sites in the suprachiasmatic nucleus (SCN) of the rat hypothalamus. Because the distribution of these binding sites had previously been reported to conform to that of both vasoactive intestinal peptide (VIP)-containing nerve cell bodies and serotonin (5-HT)-containing axons, the following experimental approaches were used: (1) the overlap between autoradiographically labeled NT binding sites and immunocytochemically labeled VIP neurons was examined in adjacent 5-microns-thick sections taken across the entire rostrocaudal extent of the SCN; and (2) the density of NT binding sites was examined by quantitative autoradiography following cytotoxic lesioning of 5-HT afferents. Double-labeling studies demonstrated precise overlap between 125I-NT binding and VIP immunostaining throughout the SCN. Moreover, at high magnification intensely VIP-immunoreactive neurons were found in direct register with 125I-NT-labeled cells visualized in adjacent sections. Densitometric autoradiographic studies demonstrated a significant reduction in specific 125I-NT binding within the SCN following intracerebroventricular injection of the neurotoxin, 5,7-dihydroxytryptamine. Taken together, these results indicate that within the SCN, NT receptors are present both presynaptically on serotonin axons and postsynaptically on the perikarya and dendrites of VIP-containing neurons.


Detection of mycobacterial DNA in pleural fluid from patients with tuberculous pleurisy by means of the polymerase chain reaction: comparison of two protocols. The detection of mycobacterial DNA in clinical samples on the basis of the polymerase chain reaction is a promising approach for the rapid diagnosis of tuberculous infections. No consensus exists, however, about which protocols are most sensitive, and the usefulness of this approach in the diagnosis of tuberculous effusions has been assessed in few patients. The sensitivity of two protocols was compared for the detection of DNA from Mycobacterium tuberculosis in samples containing known amounts of mycobacterial DNA and in DNA extracted from 15 tuberculous pleural effusions. The results obtained for pleural fluid have been compared with cytological findings and with results obtained by standard microbiological techniques. Mycobacteria could be detected by acid fast staining in none and by culture in three of the 15 pleural fluid samples. A protocol based on the detection of the IS6110 insertion element (which could detect one mycobacterial genome/sample reproducibly) gave a positive result in nine of the 15 tuberculous effusions, though some samples were only intermittently positive (p less than 0.05 compared with culture). In contrast, a protocol based on the detection of the gene coding for the 65 kD mycobacterial antigen (which could detect mycobacterial genomes only if there were at least 10/sample) gave a positive result in three of the 15 tuberculous effusions. Pleural fluid that was always positive with the amplification procedure detecting the IS6110 sequence contained more neutrophils (30% (SD 27%)) than samples that were intermittently positive or always negative (3% (3%)); mycobacterial DNA was never detected in the four samples containing less than 1% neutrophils. The amplification of the IS6110 insertion element represents a rapid and sensitive means of detecting M tuberculosis in tuberculous effusions. The enrichment of cells containing mycobacteria (possibly neutrophils) before DNA extraction may be required to improve the sensitivity of this approach.