[New aspects in the diagnosis, pathogenesis and therapy of schizophrenic negative symptoms]. The treatment of negative symptoms requires a thorough diagnostic assessment. Their lack of homogeneity has important clinical implications. Negative symptoms are unspecific for schizophrenia, differ in cause and manifestation, and may appear at all stages of the illness. There is much evidence for the responsivity of negative symptoms to pharmacological and psychosocial treatment. This paper gives a short survey of the historical development of the concept of negative symptoms, the development of scales specifically measuring both negative and positive symptoms, and various recently discussed hypotheses concerning pathogenesis and treatment strategies. Important is a therapeutic concept individually adjusted to the patient's needs, including pharmacological, psychotherapeutic and social measures.


[Pisa syndrome].

[Pisa syndrome]. The Pisa Syndrome is a rare dystonic syndrome, first described by Ekbom et al. 1972 as a side effect of neuroleptic therapy and subsequently by Patel et al. 1991 as a symptom of dementia of the Alzheimer type. It consists of tonic flexion of the trunk to one side, accompanied by a slight rotation. The presentation of three clinical cases, including one without neuroleptic but with thymoleptic medication, is followed by etiological and therapeutical considerations as well as data concerning the prevalence of this rare syndrome. Finally, the question of a possible psychogenesis of this condition is discussed.


[Adult form of metachromatic leukodystrophy with predominantly psychotic manifestations]. This is a report of one of the few cases of an adult form of metachromatic leukodystrophy with almost exclusively psychiatric symptoms. This should point out the importance of a careful organic diagnostic assessment in all patients with the first manifestation of a psychiatric disease. In this case an investigation of the patient's family confirmed the heterozygote genetic origin of the disease.


GABAergic influences on somatostatin secretion from hypothalamic neurons cultured in defined medium. The effects of GABAergic influences intrinsic to the hypothalamus on the secretion of somatostatin were studied using cultured fetal rat hypothalamic neurons. The existence of GABAergic neurons within the cultures was confirmed by immunocytochemistry. These neurons appeared to be actively secreting GABA as antagonism of GABAA receptors with bicuculline and picrotoxinin caused a dose-dependent increase in the release of immunoreactive somatostatin (SRIF), which was Ca(2+)-dependent. Although exogenous GABA inhibited SRIF secretion at concentrations of 10(-6) M and greater, muscimol, a GABAA agonist, inhibited SRIF release at 10(-8) M, whereas baclofen, a GABAB agonist, required concentrations two orders of magnitude greater to produce an effect. Phaclofen, a GABAB antagonist, was inactive (10(-8)-10(-4) M). A GABA uptake inhibitor, SKF 89976A, produced a dose-dependent inhibition of SRIF release. These results, therefore, support a role for intrahypothalamic GABA neurons in the regulation of SRIF secretion in the neonatal rat, predominantly via a type A receptor, and provide further evidence for a neuroendocrine role for GABA in controlling growth hormone secretion.


Dopaminergic regulation of luteinizing hormone-releasing hormone release at the median eminence level: immunocytochemical and physiological evidence in hens. Theoretically, the most effective inhibitory control of hypophysiotropic luteinizing hormone-releasing hormone (LHRH) release might occur through a presynaptic inhibition of LHRH neuronal terminals at the median eminence (ME) level. Since: (a) we have recently reported the existence of synaptic contacts between dopamine- and LHRH-containing processes in the ewe ME, and (b) nutritional deprivation induces an ovulatory failure in both birds and mammals, we have assessed the possibility that the anovulatory state induced by feed withdrawal (FW) in laying hens, might be caused by a dopaminergic inhibition of LHRH release at the ME level. Laying hens at the start (35 weeks old) and end (75 weeks old) of their commercial egg-laying life were killed at 0, 1, 2 and 4 days after FW. Serum luteinizing hormone (LH) and progesterone (P4), in vitro release of LHRH by isolated ME, and LHRH content in ME and preoptic area (POA) were determined by RIA. ME content of dopamine (DA) and its main metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) were assessed by LCED. The distribution of LHRH and tyrosine hydroxylase (TH)-containing processes at the ME level of the hen was determined immunocytochemically. In the hen, LHRH-containing cell bodies are localized in the anterior hypothalamus and medial POA. LHRH-containing axons project toward the ME and infundibulum through the ventral-lateral hypothalamus. TH-containing perikarya are concentrated in the arcuate nucleus and in the adjacent part of the periventricular nucleus, dorsal to the arcuate. TH-containing axons converge toward the ME and descend into the infundibulum. Dense concentrations of TH- and LHRH-containing processes are located in the lateral and mediobasal portions of the external layer of the ME, providing opportunities for synaptic interactions between them. Ovulatory failure and regression of the ovary and reproductive tract occurred 2-3 days after FW at the end, but not at the beginning of the hen's commercial egg-laying life. After FW, hens at the end of their productive life had higher (p less than 0.01) tuberoinfundibular DA turnover, produced less LHRH, and had lower serum LH and P4 than hens undergoing FW at the beginning of their productive life. In addition, in vitro release of HRH from denervated ME tissue of hens undergoing FW at the end of their commercial egg-laying life was higher and was reversed in a dose-dependent fashion by DA, but not by serotonin. Thus, the ovulatory failure associated with FW in laying hens might be caused by a presynaptic inhibition of in vivo LHRH release at the level of ME hypothalamic neuronal terminals.


Dynorphin-processing endopeptidase in the rat anterior pituitary lactotrophic cell line, GH4C1. Several peptide hormones and neurotransmitters are produced by cleavage at the monobasic processing sites. An endoprotease capable of cleaving a dynorphin peptide at the monobasic processing site is secreted from the rat anterior pituitary lactotrophic cell line, GH4C1. When characterized by fast protein liquid chromatography using an ion exchange column, the majority of the endoprotease activity elutes as a single symmetrical peak around 0.3 M NaCl. The protease inhibitor profile suggests that the activity is due to putative thiol protease. These enzymatic properties are similar to a monobasic processing enzyme previously found in bovine pituitary and in the rat brain. The secretory pathway which contains the enzyme activity in GH4C1 cells was characterized by stimulation of secretion by thyrotropin releasing hormone, forskolin, phorbol ester, or potassium chloride. The secretion of the enzyme activity was substantially increased by these compounds suggesting that the GH4C1 cells secrete the activity via the regulated pathway. A hormonal treatment of the GH4C1 cells which has been previously shown to produce a substantial increase in the number of secretory granules and ir-prolactin has been found in this study to elevate this enzyme activity 2-fold. This increase is similar to that seen in the carboxypeptidase E activity, another putative peptide hormone processing enzyme activity. These data suggest that the peptide processing activity is regulated to a small but significant extent and is coordinately regulated with carboxypeptidase E activity.


Release of endogenous amino acids, including homocysteic acid and cysteine sulphinic acid, from rat hippocampal slices evoked by electrical stimulation of Schaffer collateral-commissural fibres. This study examined the release of endogenous amino acids from acute hippocampal slices, upon stimulation of the Schaffer collateral-commissural fibres. One-minute samples of superfusate were collected via a cannula placed over the CA1 stratum radiatum, and were analysed by reversed-phase high performance liquid chromatography. Evoked potentials were recorded to ascertain stimulation efficacy. Four minutes of continuous 50 Hz stimulation produced a tetrodotoxin-sensitive release of aspartate and glycine in the second minute of stimulation, as well as a tetrodotoxin-sensitive release of cysteine sulphinic acid, during stimulation and of homocysteic acid, following stimulation. Such 50 Hz stimulation also produced a tetrodotoxin-insensitive decrease in methionine levels, but no significant changes in any of the other 15 amino acids measured. Four minutes of continuous 1 Hz stimulation produced no changes in the levels of any of the amino acids measured, but four 600-ms trains of 100 Hz stimulation, which, unlike the 1 Hz stimulation, produced long-term potentiation, resulted in significant increases in levels of cysteine sulphinic acid and homocysteic acid, but not of any of the other amino acids measured. These results suggest that aspartate, glycine, homocysteic acid, and cysteine sulphinic acid play a role in synaptic transmission in the Schaffer collateral-commissural fibres, and that cysteine sulphinic acid and homocysteic acid may be released specifically by high-frequency stimulation.


Comparison of neuronal inositol 1,4,5-trisphosphate 3-kinase and receptor mRNA distributions in the adult rat brain using in situ hybridization histochemistry. As a result of its interaction with a specific receptor, inositol 1,4,5-trisphosphate mobilizes intracellular calcium. The metabolism of inositol 1,4,5-trisphosphate is rather complex: inositol 1,4,5-trisphosphate 3-kinase produces inositol 1,3,4,5-tetrakisphosphate, a putative second messenger. In order to elucidate inositol 1,3,4,5-tetrakisphosphate function, a comparative in situ hybridization study of the distributions of inositol 1,4,5-trisphosphate 3-kinase and receptor mRNAs was performed in the adult rat brain using oligonucleotides derived from their cDNA sequences. The neuronal distributions of the mRNA for the receptor were larger than for the kinase. Highest levels of both mRNAs were found in the cerebellar Purkinje cells, where they were enriched in their neuronal perikarya and to a lesser extent in their dendrites. In addition to the cerebellum, mRNAs were mainly detected in the hippocampal pyramidal cells of the CA1 sector of the Ammon's horn and in the granule cells of the dentate gyrus, and also in a majority of the neurons in the cortical layers II-III and V, especially in the frontal cortex and cingulate cortex; caudate-putamen, accumbens, olfactory tubercle and Calleja islets; claustrum; anterior olfactory nucleus; taenia tecta; piriform cortex; dorsolateral septum; bed nucleus stria terminalis; amygdala; hippocampal CA2-4 sectors and subiculum. The inositol 1,4,5-trisphosphate receptor mRNA but not kinase mRNA was found in a majority of the neurons in the thalamus, especially in the parafascicular nucleus; hypothalamus, especially the medial hypothalamus; substantia nigra pars compacta and ventral tegmental area; superior colliculus; lateral interpeduncular nucleus and central gray. Taking into account the limitation in sensitivity of the technique, both mRNAs were not detected in glial cells and in the olfactory bulb; basal nucleus of Meynert, diagonal band nuclei; medial septal nucleus; substantia innominata; globus pallidus; entopeduncular nucleus; substantia nigra pars reticulata; ventral pallidum; subthalamic nucleus; spinal cord and dorsal root ganglia. In conclusion, cerebellum and hippocampus appear to contain almost similar levels of kinase mRNA. This is in contrast to receptor mRNA levels which were at much higher levels in the cerebellum when compared with the hippocampus. For this reason, we have chosen hippocampal CA1 pyramidal cells and dentate gyrus granule cells for studying inositol 1,4,5-trisphosphate 3-kinase function.


Neurotensin increases tyrosine hydroxylase messenger RNA-positive neurons in substantia nigra after retrograde axonal transport. In previous studies we have shown that labelled neurotensin injected into the rat striatum was found to be transported retrogradely in dopaminergic neurons through a process which was receptor and microtubule dependent. Now, we show, by in situ hybridization, the consequences of the striatal injection of neurotensin on the gene expression of tyrosine hydroxylase in the substantia nigra. Rats were injected with neurotensin or its fragments in the striatum of one side and with saline or the inactive fragment on the other. The number of nigral cells expressing tyrosine hydroxylase mRNA was found to increase by 40% after injection of neurotensin or its active fragment (neurotensin 8-13). In the same experimental conditions, the inactive fragment (neurotensin 1-8) was without effect. Time-course experiments revealed that the tyrosine hydroxylase mRNA was increased 4 h after neurotensin injection but not at 1 or 16 h. The fact that the increase of mRNA parallels the appearance of labelled neurotensin in the substantia nigra indicates that the changes in the gene expression of tyrosine hydroxylase might be the consequence of the retrograde axonal transport of neurotensin. These results represent the first evidence for the existence of a long-distance retrograde signalling process in which the neuropeptide and presumably its receptor may serve as information molecule between synapses and the cell body.


Physiological effects of chronic and acute application of N-methyl-D-aspartate and 5-amino-phosphonovaleric acid to the optic tectum of Rana pipiens frogs. Visually elicited activity contributes to the formation of orderly connections in the optic tectum of frogs. Glutamate receptors of the N-methyl-D-aspartate class participate in this process. Blocking those receptors interferes with activity-dependent refinement of maps in normal frogs and of ocular dominance bands in surgically produced animals with three eyes. Chronic application of N-methyl-D-aspartate sharpens the bands. The possibility that 5-amino-phosphonovaleric acid depresses tectal responsiveness was motivation for studying the effects of 5-amino-phosphonovaleric acid and N-methyl-D-aspartate applied both chronically and acutely. We evaluated tectal responsiveness to visual input by presenting flashes of light to one eye and recording responses in the ipsilateral tectal lobe. This method reveals the output of the tectal cells contralateral to the stimulated eye. These cells project via the nucleus isthmi to the opposite tectal lobe. We also mapped the receptive field dimensions of the crossed isthmotectal axons. Our results show that acute topical application of 500 microM or 1 mM N-methyl-D-aspartate dramatically increases spontaneous activity, while 100 microM N-methyl-D-aspartate causes little change. Chronic treatment with N-methyl-D-aspartate at a low dose (estimated to be in the micromolar range) shown to influence retinotectal mapping, reduces response latencies but produces no statistically significant changes in tectal cell firing rates or receptive field size. Acute application of 5-amino-phosphonovaleric acid produces complex results: 10 microM produces no changes in firing, 100 microM 5-amino-phosphonovaleric acid decreases firing, and doses of 500-100 microM increase the firing.(ABSTRACT TRUNCATED AT 250 WORDS)


Peroxide effects on [3H]L-glutamate release by synaptosomes isolated from the cerebral cortex. Basal (non-depolarized) and high K(+)-stimulated [3H]L-glutamate release in the presence and absence of Ca2+ were assessed using presynaptic nerve terminals (synaptosomes) isolated from the cerebral cortex of the guinea pig. Basal glutamate release was found to be Ca(2+)-independent and was significantly increased following treatment with hydrogen peroxide (H2O2). On the other hand, depolarization-induced release had both a Ca(2+)-dependent and Ca(2+)-independent component. Both components of stimulated release were suppressed by H2O2. In fact, Ca(2+)-dependent evoked release was virtually eliminated by H2O2 pretreatment. The data suggest that H2O2 exerts a differential effect on the neurochemical mechanisms involved in basal and stimulated glutamate release at the presynaptic nerve terminal.


Tau antigenic changes induced by glutamate in rat primary culture model: a biochemical approach. Primary neuronal cultures were treated with glutamate to induce an increase of Tau immunoreactivity similar to that observed in Alzheimer's disease. The Tau profile of neurones in culture before and after exposure to glutamate was analyzed on immunoblots with anti-Tau, anti-paired helical filaments (PHF) and antibody specific for modified Tau. Differences were observed between treated and control cultures: glutamate induced a shift of immunodetection from the lowest to the highest molecular weight Tau isoform and an acidification of Tau proteins. However, these modifications are not exactly those observed in Alzheimer's disease since we were not able to detect 'Alzheimer-type' epitopes on Tau proteins after the glutamate exposure.


Pertussis toxin pretreatment abolishes the inhibitory effect of riluzole and carbachol on D-[3H]aspartate release from cultured cerebellar granule cells. The release of D-[3H]aspartate from cultured cerebellar granule cells evoked by glutamic acid can be inhibited by riluzole and the muscarinic agonist carbachol. The combined application of maximally efficacious concentrations of riluzole and carbachol produces no greater inhibition than that seen with either agent alone, indicating that a common mechanism is involved. The effects of both agents are abolished when the cells have been pretreated with pertussis toxin, which suggests that this mechanism may involve a GTP-binding protein. The effect of pertussis toxin pretreatment is not mimicked by cholera toxin, nor does pertussis toxin pretreatment interfere with the inhibitory effect of the competitive excitatory amino acid receptor antagonist D-alpha-aminoadipic acid.


Characterization of a paired box- and homeobox-containing quail gene (Pax-QNR) expressed in the neuroretina. The retina is an integral part of the central nervous system, and consists of two layers, the outer pigmented layer and the inner sensory layer or neuroretina (NR). The NR layer contains several strata of cells (glial and neuronal) derived from proliferating neuroectodermal precursors that differentiate after terminal mitosis. In vitro, NR cells can differentiate not only into neuronal and glial types, but also into pigment and lens cells. Quail (Coturnix coturnix japonica) NR cells (QNR) infected with MC29 transforming retrovirus become pigmented after several passages in vitro. In order to characterize the genes expressed in these pigmented MC29 QNR, a cDNA library was prepared from these cells. After differential screening we have isolated a cDNA clone which identifies an RNA expressed in NR but not in the pigmented layer of the retina. This cDNA encodes a protein related to that of Drosophila, mouse and zebrafish paired box- and homeobox-containing segmentation genes and is called Pax-QNR. The expression of Pax-QNR in the NR is confined to the ganglionic cell layer and to the lower part of the inner nuclear layer containing the amacrine or correlation neurones.


Transient expression of the Epstein-Barr virus LMP1 gene in human primary B cells induces cellular activation and DNA synthesis. The Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) and Epstein-Barr virus nuclear antigen 2 (EBNA2) are expressed in EBV-immortalized human B cells. It has previously been shown that transfection of the LMP1 and EBNA2 genes into Burkitt's lymphoma cell lines results in the up-regulation of CD23, CD21, ICAM-1 and LFA-1 cell-surface proteins. In the present study, the effects of transient expression of the LMP1 and EBNA2 genes were studied in normal primary human B cells pretreated with UV-inactivated EBV particles. To identify and purify cells which express the transfected DNA we used a gene encoding a surface molecule, CD2, as a co-transfection marker. We show that transient expression of the LMP1 gene, from heterologous promoters, is sufficient to induce cellular enlargement and up-regulation of surface expression of the activation markers CD23, CD21, ICAM-1 and LFA-1 in primary B cells. Most importantly, we show that transient expression of the LMP1 gene is sufficient to induce DNA synthesis in human primary B cells. Transient EBNA2 expression enhanced the effect of transient LMP1 expression on CD21 and CD23 cell-surface expression but, under our experimental conditions, inhibited the induction of DNA synthesis by LMP1. We conclude that activation of primary B cells with inactivated EBV particles, followed by transient expression of only two viral genes, EBNA2 and LMP1, is sufficient to reconstitute some of the early events of B-cell immortalization by EBV.


Transformation mediated by the human HER-2 gene independent of the epidermal growth factor receptor. Amplification of the HER-2 (c-erbB-2) gene and overexpression of the p185HER-2 gene product is found in approximately one-third of primary human breast and ovarian cancers and is associated with a poor clinical outcome of early relapse and death. The HER-2 gene encodes a cell-surface growth factor receptor with intrinsic tyrosine kinase activity. Wild-type human HER-2 has been shown to act as a potent oncogene when over-expressed in mouse fibroblasts. Recent data suggest that the mechanism by which HER-2 mediates transformation requires the interaction of the epidermal growth factor (EGF) receptor. To test whether overexpression of normal human HER-2 can transform cells independently of the EGF receptor, we have introduced multiple copies of HER-2 into the EGF receptor-negative cell line, NR6, and have performed assays for both transformation and tumorigenicity. Engineered NR6 cells that overexpress the HER-2 gene product display a highly transformed and tumorigenic phenotype as compared with control cells. Additionally, a monoclonal antibody to the extracellular domain of the HER-2 receptor is able to inhibit the proliferation of the overexpressing cells in vitro as well as tumor growth in vivo. This study provides clear evidence that HER-2-mediated transformation can be achieved independently of the EGF receptor.


Incomplete stroma formation after allogeneic marrow or autologous blood stem cell transplantation. Long term and semi-solid culture techniques were used to evaluate the quality of stroma produced by bone marrow from 33 normal subjects and 57 patients (46 allogeneic bone marrow and 11 autologous blood stem cell transplant recipients). Bone marrow from transplant recipients was capable of sustained CFU-GM and nucleated cell production in long term culture. However, only 13% of the marrow investigated developed a complete, confluent stromal layer. These stromal abnormalities were observed in spite of complete hematopoietic reconstitution after transplantation and rarely improved with time. Our results suggest that the hematopoietic microenvironment is very fragile and susceptible to long term damage as a result of chemotherapy and the conditioning regimes used prior to transplantation.


Carboxyl-terminal deletion and point mutations decrease the transforming potential of the activated rat neu oncogene product. The rat neu oncogene encodes a constitutively activated growth factor receptor/transmembrane tyrosine kinase, p185Tneu, that is structurally similar to yet distinct from the epidermal growth factor receptor. To explore the role of the carboxyl-terminal region and of putative autophosphorylation sites in regulating the activity of the rat p185Tneu (T, transforming) protein, we used site-directed mutagenesis to generate a p185Tneu mutant in which a putative tyrosine autophosphorylation site (residue 1253) at the extreme carboxyl terminus was replaced by a phenylalanine residue and a mutant in which the carboxyl-terminal 122 amino acids were deleted. These proteins were expressed in NIH 3T3 cells at comparable levels and exhibited similar autophosphorylation activity, exogenous substrate phosphorylation ability, oligomerization levels, and responsiveness to a partially purified neu-activating factor. However, the mutant p185Tneu proteins displayed a decreased transforming capacity both in vitro and in vivo. This analysis demonstrated that the carboxyl-terminal domain and at least one putative tyrosine autophosphorylation site of p185Tneu play a role in positively regulating the cell growth-regulating properties of the neu protein.


Prolactin release from perifused human decidual explants; effects of decidual prolactin-releasing factor (PRL-RF) and prolactin release-inhibitory factor (PRL-IF). The dynamics of prolactin release from human decidual explants were studied under basal conditions, in response to decidual prolactin-releasing factor (PRL-RF), and in response to PRL-RF in the presence of decidual prolactin release-inhibitory factor (PRL-IF) or other factors known to inhibit prolactin release in static cultures. Explants were perifused with medium at a rate of 6 ml/h, and the medium was collected at 5 min intervals. The explants released prolactin for up to 20 h without evidence of cell necrosis, with the rate of prolactin decreasing gradually from 3.9 +/- 0.1 ng/5 min during the first 2 h to 2.2 +/- 0.1 ng/5 min during the last 2 h of exposure. PRL-RF, a 23.5 KMr protein released by the placenta, stimulated a dose-dependent increase in prolactin release from the perifused explants that occurred within the first 5 min of exposure and persisted until the exposure to the releasing factor was discontinued. PRL-IF, a 35-45 K Mr protein released by the decidua, caused a dose-dependent inhibition of PRL-RF-mediated prolactin release. Dibutyryl cAMP, cholera toxin, sn-1, 2-dioctonylglycerol, PMA, and arachidonic acid, which inhibit basal prolactin release from static decidual cultures, also caused a dose-dependent inhibition of prolactin release in response to PRL-RF. In each instance, the maximal dose of the agents tested inhibited PRL-RF-mediated prolactin release by greater than 84 per cent. These results indicate that the stimulation of prolactin by PRL-RF is inhibited by PRL-IF and pharmacologic agents that inhibit basal prolactin release.(ABSTRACT TRUNCATED AT 250 WORDS)


Segmental determination in Drosophila conferred by hunchback (hb), a repressor of the homeotic gene Ultrabithorax (Ubx). The activity of homeotic genes in Drosophila cells determines segment-specific morphogenesis. Here, we provide evidence that the product of hunchback (hb), a segmentation gene, acts as a direct repressor or "silencer" of the homeotic gene Ultrabithorax (Ubx) and thus prevents ectopic activity of this gene: we show, by stable integration of reporter gene constructs, that hb protein binding sites are capable of repressing at a distance the activity of an embryonic Ubx enhancer outside the Ubx expression domain. This silencing activity is observed at advanced embryonic stages, at a time when the hb gene product is no longer detectable or required, and is dependent on the function of Polycomb (Pc). We propose a working hypothesis as to how hb protein in a "hit-and-run" fashion may effect stable and heritable silencing of the Ubx gene throughout advanced stages of development, thus mediating repression of this homeotic gene outside its realm of function.


Multiple kinetic states underlying macroscopic M-currents in bullfrog sympathetic neurons. M-current is a time- and voltage-dependent potassium current which is suppressible by muscarinic receptor activation. We have used curve fitting and noise analysis to determine if macroscopic M-currents deviate from a previously predicted simple two-state kinetic scheme. The M-current was best described by three kinetically distinct components: 'fast' (tau 0), 'intermediate' (tau 1) and 'slow' (tau 2) time constants. The 'fast' (tau 0) and 'intermediate' (tau 1) components were identified from the spectra of M-current noise at potentials positive to the cells' resting membrane potential. The 'intermediate' (tau 1) and 'slow' (tau 2) components were seen by curve fitting M-current deactivation currents. The 'intermediate' (tau 1) time constant was voltage dependent (decreasing e-fold in 23 mV), but voltage dependence of the 'fast' (tau 0) and 'slow' (tau 2) components was not obvious. All kinetic components were sensitive to muscarine, with the 'intermediate' (tau 1) and 'slow' (tau 2) being equally so. These data suggest that all components may derive from the same channel population, and that the M-channel may have at least four kinetic states.


Regional mapping of prion proteins in brain. Scrapie is characterized by the accumulation of a protease-resistant isoform of the prion protein PrPSc. Limited proteolysis and chaotropes were used to map the distribution of PrPSc in cryostat sections of scrapie-infected brain blotted onto nitrocellulose membranes, designated histoblots. Proteolysis was omitted in order to map the cellular isoform of the prion protein (PrPC) in uninfected brains. Compared with immunohistochemistry, histoblots increased the sensitivity for PrPSc detection and showed different patterns of PrPSc accumulation. In Syrian hamsters with Sc237 scrapie, the most intense PrPSc signals occurred in sites with relatively little PrPC, suggesting that aberrant localization of prion protein may be an important feature in the pathogenesis of prion diseases. Immunostaining of PrPSc in white-matter tracts suggested that prions spread along neuroanatomical pathways. PrPSc immunostaining in histoblots was quantitated by densitometry, permitting assessment of the extent of PrPSc accumulation within specific structures. Histoblots were also useful in localizing PrPCJD and beta/A4-amyloid peptide in the brains of patients with Creutzfeldt-Jakob disease and Alzheimer disease, respectively.


Putative immunolocalization of the mechanoelectrical transduction channels in mammalian cochlear hair cells. Hair cells bear an apical bundle of stereocilia arranged in serried rows. Deflection of the bundle controls the opening and closing of mechanoelectrical transduction channels, thereby altering the conductance across the apical plasma membrane. Two locations for these channels have been proposed in the bundle, either near the bases of the stereocilia or towards their tips. One hypothesis that is consistent with the latter possibility suggests that fine extracellular filaments, which run between the tips of the shorter stereocilia and the sides of the taller stereocilia behind, operate the channels. Determining the precise position of the channels is essential to test this hypothesis. We have therefore attempted to localize them immunocytochemically. Because hair-cell transduction is amiloride sensitive, the channels may have an amiloride-binding site associated with them. We have therefore used a polyclonal antibody raised against another amiloride-sensitive ion channel to hunt for them. This antibody recognizes a 62-64 kDa band in immunoblots of cochlear tissue, and produces discrete labelling in the hair bundle. This is most concentrated just below the tips of the shorter stereocilia, coinciding with a region of specialization in the closely apposed membranes of the short and tall stereocilia but not with either end of the tip link.


Motor practice and neurophysiological adaptation in the cerebellum: a positron tomography study. We have used positron tomography (PET) to demonstrate that some parts of the motor system exhibit physiological adaptation during the repeated performance of a simple motor task, but others do not. In contrast to the primary sensori-motor cortex, the cerebellum exhibits a decrease in physiological activation (increases in regional blood flow during performance) with practice. A new application of factorial experimental design to PET activation studies was used to make these measurements in four normal males. This design allowed adaptation to be examined by testing for an interaction between regional cerebral blood flow (rCBF) increases brought about by a motor task and the number of trials (time). These findings are interpreted as the neurophysiological correlates of synaptic changes in the cerebellum associated with motor learning in man.


Error-prone signalling.

Error-prone signalling. The handicap principle of Zahavi is potentially of great importance to the study of biological communication. Existing models of the handicap principle, however, make the unrealistic assumption that communication is error free. It seems possible, therefore, that Zahavi's arguments do not apply to real signalling systems, in which some degree of error is inevitable. Here, we present a general evolutionarily stable strategy (ESS) model of the handicap principle which incorporates perceptual error. We show that, for a wide range of error functions, error-prone signalling systems must be honest at equilibrium. Perceptual error is thus unlikely to threaten the validity of the handicap principle. Our model represents a step towards greater realism, and also opens up new possibilities for biological signalling theory. Concurrent displays, direct perception of quality, and the evolution of 'amplifiers' and 'attenuators' are all probable features of real signalling systems, yet handicap models based on the assumption of error-free communication cannot accommodate these possibilities.


Response to fibronectin-integrin interaction in leukaemia cells: delayed enhancing of a K+ current. In murine erythroleukaemia cells, the response of ion channels was followed before and after contact with fibronectin-coated latex microspheres. Patch-clamp experiments in 'whole-cell' and in 'cell-attached' configurations showed that cell adhesion to fibronectin promoted plasma membrane hyperpolarization mediated by activation of potassium channels that were indistinguishable from calcium-dependent potassium channels K(Ca) in these cells. K+ current increase began in 5-6 min and was completed about 10 min after the first contact. The timecourse of this process recorded from 'whole-cell' was very similar to that followed in intact cells by observing the increase of single channel currents. The open probability of single channels in the patch increased after contact, revealing that this activation is propagated at distance from the adhesion site. The slow onset of the effect suggests the presence of a complex regulatory pathway between fibronectin-integrin binding and activation of potassium channels. Decreasing cytoplasmic free Ca2+ concentration to pCa 9 diminished, but did not inhibit, the response. The current induced by fibronectin was not blocked by apamin, alpha-charybdotoxin or glibenclamide, but was abolished by high concentrations of tetraethylammonium (TEA). These data suggest for the first time the existence of a specific regulative connection between integrin receptors and ionic channels.


Interaction of avidin with the lipoyl domains in the pyruvate dehydrogenase multienzyme complex: three-dimensional location and similarity to biotinyl domains in carboxylases. Avidin can form intermolecular cross-links between particles of the pyruvate dehydrogenase multienzyme complex from various sources. Avidin does this by binding to lipoic acid-containing regions of the dihydrolipoamide acetyltransferase polypeptide chains that comprise the structural core of the complex. It is inferred that the lipoyl domains of the acetyltransferase chain extend outwards from the interior of the enzyme particle, interdigitating between the subunits of the other two enzymes bound peripherally in the assembled structure, with the lipoyl-lysine residues capable of reaching to within at least 1-2 nm of the outer surface of the enzyme complex (diameter ca. 37 nm). The distribution of enzymic activities between different domains of the dihydrolipoamide acetyltransferase chain implies that considerable movement of the lipoyl domains is a feature of the catalytic activity of the enzyme complex. There is evidence that the lipoyl domain of the 2-oxo acid dehydrogenase complexes is similar in structure to a domain that binds the cofactor biotin, also in amide linkage with a specific lysine residue, in the biotin-dependent class of carboxylases.


Oncogenes, anti-oncogenes and the immune response to cancer: a mathematical model. We develop a mathematical model for the initial growth of a tumour after a mutation in which either an oncogene is expressed or an anti-oncogene (i.e. tumour suppressor gene) is lost. Our model incorporates mitotic control by several biochemicals, with quite different regulatory characteristics, and we consider mutations affecting the cellular response to these control mechanisms. Our mathematical representation of these mutations reflects the current understanding of the roles of oncogenes and anti-oncogenes in controlling cell proliferation. Numerical solutions of our model, for biologically relevant parameter values, show that the different types of mutations have quite different effects. Mutations affecting the cell response to chemical regulators, or resulting in autonomy from such regulators, cause an advancing wave of tumour cells and a receding wave of normal cells. By contrast, mutations affecting the production of a mitotic regulator cause a slow localized increase in the numbers of both normal and mutant cells. We extend our model to investigate the possible effects of an immune response to cancer by including a first order removal of mutant cells. When this removal rate exceeds a critical value, the immune system can suppress tumour growth; we derive an expression for this critical value as a function of the parameters characterizing the mutation. Our results suggest that the effectiveness of the immune response after an oncogenic mutation depends crucially on the way in which the mutation affects the biochemical control of cell division.


Amplification and rearrangement of a prokaryotic metallothionein locus smt in Synechococcus PCC 6301 selected for tolerance to cadmium. Metal-tolerant cyanobacteria have been isolated from metal-polluted aquatic environments and also selected in culture, but no genes which confer metal tolerance have been described. To investigate the possibility that amplification of a prokaryotic metallothionein gene (smtA), or rearrangement of the smt locus, could be involved in the development of Cd tolerance in Synechococcus PCC 6301, Cd-tolerant lines were selected by stepwise adaptation of a Synechococcus culture. An increase in smtA gene copy number and the appearance of unique additional smtA restriction fragments (both larger and smaller) were detected in these tolerant lines (tolerant to 0.8 microM Cd, 1.3 microM Cd and 1.7 microM Cd). Stepwise adaptation was repeated by using a culture of Synechococcus PCC 6301 inoculated from a single plated colony to obtain four new lines (tolerant to 1.4 microM Cd, 1.8 microM Cd, 2.6 microM Cd and 3.2 microM Cd). Amplification of the smtA gene and development of unique smtA restriction fragments (larger and smaller) were once again detected in these tolerant lines. Amplification and rearrangement of the smt locus were only detected in the seven Cd-tolerant lines, with no evidence of amplification or rearrangement in the non-tolerant lines from which they were derived. As a control, another gene, psaE, was also monitored in these cell lines. There was no evidence of amplification or rearrangement of psaE in the non-tolerant or any of the Cd-tolerant lines.


Isolation of deacetoxycephalosporin C from fermentation broths of Penicillium chrysogenum transformants: construction of a new fungal biosynthetic pathway. Deacetoxycephalosporin C (DAOC), a precursor of cephalosporins excreted by Cephalosporium and Streptomyces species, has been produced in Penicillium chrysogenum transformed with DNA containing a hybrid penicillin N expandase gene (cefEh) and a hybrid isopenicillin N epimerase gene (cefDh). DAOC from a P. chrysogenum transformant was identified by ultraviolet light (UV), high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) and mass spectrum analyses. P. chrysogenum transformed with DNA containing cefEh without cefDh did not produce DAOC. Untransformed P. chrysogenum produced penicillin V (phenoxymethylpenicillin) but not DAOC. Transformants also produced penicillin V but, in general, less than untransformed P. chrysogenum. The cefEh and cefDh genes were constructed by replacing the open reading frame (ORF) of cloned P. chrysogenum pcbC and penDE genes with the ORF of the Streptomyces clavuligerus expandase gene, cefE, and the ORF of the Streptomyces lipmanii epimerase gene, cefD, respectively. Analyses of representative transformants suggested that production of DAOC occurred via cefEh and cefDh genes stably integrated in the P. chrysogenum genome. DNA from untransformed P. chrysogenum did not hybridize to cefE or cefD gene probes.


Dichromats detect colour-camouflaged objects that are not detected by trichromats. To explain the surprisingly high frequency of congenital red-green colour blindness, the suggestion has been made that dichromats might be at an advantage in breaking certain kinds of colour camouflage. We have compared the performance of dichromats and normal observers in a task in which texture is camouflaged by colour. The texture elements in a target area differed in either orientation or size from the background elements. In one condition, the texture elements were all of the same colour; in the camouflage condition they were randomly coloured red or green. For trichromats, it proved to be more difficult to detect the target region in the camouflage condition, even though colour was completely irrelevant to the task. Dichromats (n = 7) did not show this effect, and indeed performed better than trichromats in the camouflage condition. We conclude that colour can interfere with segregation based upon texture, and that dichromats are less susceptible to such interference.


Auditory processing of complex sounds: an overview. The past 30 years has seen a remarkable development in our understanding of how the auditory system--particularly the peripheral system--processes complex sounds. Perhaps the most significant has been our understanding of the mechanisms underlying auditory frequency selectivity and their importance for normal and impaired auditory processing. Physiologically vulnerable cochlear filtering can account for many aspects of our normal and impaired psychophysical frequency selectivity with important consequences for the perception of complex sounds. For normal hearing, remarkable mechanisms in the organ of Corti, involving enhancement of mechanical tuning (in mammals probably by feedback of electro-mechanically generated energy from the hair cells), produce exquisite tuning, reflected in the tuning properties of cochlear nerve fibres. Recent comparisons of physiological (cochlear nerve) and psychophysical frequency selectivity in the same species indicate that the ear's overall frequency selectivity can be accounted for by this cochlear filtering, at least in bandwidth terms. Because this cochlear filtering is physiologically vulnerable, it deteriorates in deleterious conditions of the cochlea--hypoxia, disease, drugs, noise overexposure, mechanical disturbance--and is reflected in impaired psychophysical frequency selectivity. This is a fundamental feature of sensorineural hearing loss of cochlear origin, and is of diagnostic value. This cochlear filtering, particularly as reflected in the temporal patterns of cochlear fibres to complex sounds, is remarkably robust over a wide range of stimulus levels. Furthermore, cochlear filtering properties are a prime determinant of the 'place' and 'time' coding of frequency at the cochlear nerve level, both of which appear to be involved in pitch perception. The problem of how the place and time coding of complex sounds is effected over the ear's remarkably wide dynamic range is briefly addressed. In the auditory brainstem, particularly the dorsal cochlear nucleus, are inhibitory mechanisms responsible for enhancing the spectral and temporal contrasts in complex sounds. These mechanisms are now being dissected neuropharmacologically. At the cortical level, mechanisms are evident that are capable of abstracting biologically relevant features of complex sounds. Fundamental studies of how the auditory system encodes and processes complex sounds are vital to promising recent applications in the diagnosis and rehabilitation of the hearing impaired.


Basilar membrane responses to two-tone and broadband stimuli. The responses to sound of mammalian cochlear neurons exhibit many nonlinearities, some of which (such as two-tone rate suppression and intermodulation distortion) are highly frequency specific, being strongly tuned to the characteristic frequency (CF) of the neuron. With the goal of establishing the cochlear origin of these auditory-nerve nonlinearities, mechanical responses to clicks and to pairs of tones were studied in relatively healthy chinchilla cochleae at a basal site of the basilar membrane with CF of 8-10 kHz. Responses were also obtained in cochleae in which hair cell receptor potentials were reduced by systemic furosemide injection. Vibrations were recorded using either the Mössbauer technique or laser Doppler-shift velocimetry. Responses to tone pairs contained intermodulation distortion products whose magnitudes as a function of stimulus frequency and intensity were comparable to those of distortion products in cochlear afferent responses. Responses to CF tones could be selectively suppressed by tones with frequency either higher or lower than CF; in most respects, mechanical two-tone suppression resembled rate suppression in cochlear afferents. Responses to clicks displayed a CF-specific compressive nonlinearity, similar to that present in responses to single tones, which could be profoundly and selectively reduced by furosemide. The present findings firmly support the hypothesis that all CF-specific nonlinearities present in the auditory nerve originate in analogous phenomena of basilar membrane vibration. However, because of their lability, it is almost certain that the mechanical nonlinearities themselves originate in outer hair cells.


Sensory transduction and frequency selectivity in the basal turn of the guinea-pig cochlea. Receptor potentials recorded from outer hair cells (OHC) and inner hair cells (IHC) in the basal high-frequency turn were compared. The DC component of the IHC receptor potential is maximized to ensure that IHCS can signal a voltage response to high-frequency tones. The OHC DC component is minimized so that OHCS transduce in the most sensitive region of their operating range. The phase and magnitude of OHC receptor potentials were recorded as an indicator of the magnitude and phase of the energy which is fed back to the basilar membrane to provide the basis for the sharp tuning and fine sensitivity of the cochlea to tones. IHC receptor potentials were recorded to assess the net effect of the feedback on the mechanics of the cochlea. It was concluded that OHCS generate feedback which enhances the IHC responses only at the best frequency. At frequencies below CF, IHC DC responses are elicited only when the OHC AC responses begin to saturate.


Evidence that adaptation of suppression cannot account for auditory enhancement or enhanced forward masking. Delaying the onset of a signal relative to the onset of a simultaneous notched masker often improves the ability of listeners to 'hear out' the signal at both threshold and suprathreshold levels. Viemeister & Bacon (J. acoust. Soc. Am., 71, 1502-1507 (1982)) suggested that such auditory enhancement effects could be accounted for if the suppression produced by the masker on the signal frequency adapted, thereby releasing the signal from suppression. In support of their hypothesis, Viemeister & Bacon reported that a masker preceded by an enhancer having no component at the signal frequency produced more forward masking than did the masker by itself. Here evidence is provided from five new experiments showing that adaptation of psychophysical two-tone suppression is inadequate to account either for auditory enhancement effects or for the enhanced forward masking demonstrated by Viemeister & Bacon.


Masking release for gap detection.

Masking release for gap detection. In random noise, masking is influenced almost entirely by noise components in a narrow band around the signal frequency. However, when the noise is not random, but has a modulation pattern which is coherent across frequency, noise components relatively remote from the signal frequency can actually produce a release from masking. This masking release has been called comodulation masking release (CMR). The present research investigated whether a similar release from masking occurs in the analysis of a suprathreshold signal. Specifically, the ability to detect the presence of a temporal gap was investigated in conditions which do and do not result in CMR for detection threshold. Similar conditions were investigated for the masking level difference (a binaural masking release phenomenon). The results indicated that suprathreshold masking release for gap detection occurred for both the masking-level difference (MLD) and for CMR. However, masking release for gap detection was generally smaller than that obtained for detection threshold. The largest gap detection masking release effects obtained corresponded to relatively low levels of stimulation, where gap detection was relatively poor.


Modulation discrimination interference and auditory grouping. The detection of a change in the modulation pattern of a (target) carrier frequency, fc (for example a change in the depth of amplitude or frequency modulation, AM or FM) can be adversely affected by the presence of other modulated sounds (maskers) at frequencies remote from fc, an effect called modulation discrimination interference (MDI). MDI cannot be explained in terms of interaction of the sounds in the peripheral auditory system. It may result partly from a tendency for sounds which are modulated in a similar way to be perceptually 'grouped', i.e. heard as a single sound. To test this idea, MDI for the detection of a change in AM depth was measured as a function of stimulus variables known to affect perceptual grouping, namely overall duration and onset and offset asynchrony between the masking and target sounds. In parallel experiments, subjects were presented with a series of pairs of sounds, the target alone and the target with maskers, and were asked to rate how clearly the modulation of the target could be heard in the complex mixture. The results suggest that two factors contribute to MDI. One factor is difficulty in hearing a pitch corresponding to the target frequency. This factor appears to be strongly affected by perceptual grouping. Its effects can be reduced or abolished by asynchronous gating of the target and masker. The second factor is a specific difficulty in hearing the modulation of the target, or in distinguishing that modulation from the modulation of other sounds that are present. This factor has effects even under conditions promoting perceptual segregation of the target and masker.


The psychophysics of concurrent sound segregation. To perceptually separate concurrent complex sounds, normally hearing listeners simultaneously combine information across a wide range of frequency components. Three psychoacoustical experiments are described which investigate different forms of this across-frequency processing. The first two experiments investigate the role of coherence of frequency modulation (FM) between widely separated frequency components of a complex sound. The first experiment bolsters existing evidence that, for harmonic sounds, listeners can discriminate coherent from incoherent FM, but only by detecting the mistuning that arises from incoherent FM. The second demonstrates that, for inharmonic sounds, coherence of FM has no effect on the phenomenon of modulation detection interference (see Moore & Shailer, this symposium) once within-channel cues (combination tones and beating) are masked by background noise. It is concluded that there is not an across-frequency mechanism specific to the detection of FM incoherence. The third experiment investigates the extent to which the detection of mistuning of one component of a harmonic complex is impaired by an interfering sound (the 'interferer') with a frequency spectrum similar to that of the mistuned component. When the interferer is gated on and off with the harmonic complex, it has only a small effect provided that its level is more than 3 dB below that of the target. However, when the interferer starts before and ends after the complex, thresholds are elevated more, and this elevation occurs even for low-level interferers. Explanations of this effect in terms of adaptation and of auditory streaming are discussed.


Auditory segregation of competing voices: absence of effects of FM or AM coherence. Four experiments sought evidence that listeners can use coherent changes in the frequency or amplitude of harmonics to segregate concurrent vowels. Segregation was not helped by giving the harmonics of competing vowels different patterns of frequency or amplitude modulation. However, modulating the frequencies of the components of one vowel was beneficial when the other vowel was not modulated, provided that both vowels were composed of components placed randomly in frequency. In addition, staggering the onsets of the two vowels, so that the amplitude of one vowel increased abruptly while the amplitude of the other was stationary, was also beneficial. Thus, the results demonstrate that listeners can group changing harmonics and can segregate them from stationary harmonics, but cannot use coherence of change to separate two sets of changing harmonics.


Temporal information in speech: acoustic, auditory and linguistic aspects. The temporal properties of speech appear to play a more important role in linguistic contrasts than has hitherto been appreciated. Therefore, a new framework for describing the acoustic structure of speech based purely on temporal aspects has been developed. From this point of view, speech can be said to be comprised of three main temporal features, based on dominant fluctuation rates: envelope, periodicity, and fine-structure. Each feature has distinct acoustic manifestations, auditory and perceptual correlates, and roles in linguistic contrasts. The applicability of this three-featured temporal system is discussed in relation to hearing-impaired and normal listeners.


Pitch related to spectral edges of broadband signals. A complex tone often evokes a pitch sensation associated with its extreme spectral components, besides the holistic pitch associated with its fundamental frequency. We studied the edge pitch created at the upper spectral edge of complexes with a low-pass spectrum by asking subjects to adjust the frequency of a sinusoidal comparison tone to the perceived pitch. Measurements were performed for different values of the fundamental frequency and of the upper frequency of the complex as well as for three different phase relations of the harmonic components. For a wide range of these parameters the subjects could adjust the comparison tone with a high accuracy, measured as the standard deviation of repeated adjustments, to a frequency close to the nominal edge frequency. The detailed dependence of the matching accuracy on temporal parameters of the harmonic complexes suggests that the perception of the edge pitch in harmonic signals is related to the temporal resolution of the hearing system. This resolution depends primarily on the time constants of basilar-membrane filters and on additional limitations due to neuronal processes.


Perception of timbral analogies.

Perception of timbral analogies. Recent studies have investigated the structure of perceptual relations among musical instrument timbres by multidimensional scaling (MDS) techniques. These studies have employed both acoustically produced tones and digitally synthesized imitations and hybrids of acoustic instrument tones. The analyses of dissimilarity ratings for all pairs of a set of tones are usually represented as geometrical structures in a two- or three-dimensional Euclidean space in which the shared 'perceptual' axes are shown to have a qualitative correspondence to acoustic properties such as spectral energy distribution, onset characteristics and degree of change in spectral distribution over the duration of the tone. The present study took as a point of departure a MDS analysis for complex, synthetic tones with the aim of testing whether musician and non-musician listeners used the relations defined by the perceptual space to perform an analogies task of the sort: timbre A is to timbre B as timbre C is to which of two possible timbres, D or D'? A parallelogram model was used to select the D timbres: if the relation between A and B is represented as a vector with both magnitude and direction components, then the appropriate D should form a vector with C having similar magnitude and direction in the timbre space. Aside from conceptual difficulties with the task for both non-musicians and composers, choices for both groups provide support for the parallelogram model indicating a capacity in listeners to perceive abstract relations among the timbres of complex sounds without specific training in such a task.


Some new pitch paradoxes and their implications. This paper explores two new paradoxical sound patterns. The tones used to produce these patterns consist of six octave-related harmonics, whose amplitudes are scaled by a bell-shaped spectral envelope; these tones are clearly defined in terms of pitch class (C, C#, D, and so on) but are poorly defined in terms of height. One pattern consists of two tones that are separated by a half-octave. It is heard as ascending when played in one key, yet as descending when played in a different key. Further, when the pattern is played in any one key it is heard as ascending by some listeners but as descending by others (the tritone paradox). Another pattern that consists of simultaneous pairs of tones displays related properties (the semitone paradox). It is shown that the way the tritone paradox is perceived correlates with the speech characteristics of the listener, including his or her linguistic dialect. The findings suggest that the same, culturally acquired representation of pitch classes influences both speech production and also perception of this musical pattern.


Coding of envelope modulation in the auditory nerve and anteroventral cochlear nucleus. We have investigated responses of the auditory nerve fibres (ANFS) and anteroventral cochlear nucleus (AVCN) units to narrowband 'single-formant' stimuli (SFSS). We found that low and medium spontaneous rate (SR) ANFS maintain greater amplitude modulation (AM) in their responses at high sound levels than do high SR units when sound level is considered in dB SPL. However, this partitioning of high and low SR units disappears if sound level is considered in dB relative to unit threshold. Stimuli with carrier frequencies away from unit best frequency (BF) were found to generate higher AM in responses at high sound levels than that observed even in most low and medium SR units for stimuli with carrier frequencies near BF. AVCN units were shown to have increased modulation depth in their responses when compared with high SR ANFS with similar BFS and to have increased or comparable modulation depth when compared with low SR ANFS. At sound levels where AM almost completely disappears in high SR ANFS, most AVCN units we studied still show significant AM in their responses. Using a dendritic model, we investigated possible mechanisms of enhanced AM in AVCN units, including the convergence of inputs from different SR groups of ANFS and a postsynaptic threshold mechanism in the soma.


Modelling the sensitivity of cells in the anteroventral cochlear nucleus to spatiotemporal discharge patterns. This study investigates a potential mechanism for the processing of acoustic information that is encoded in the spatiotemporal discharge patterns of auditory nerve (AN) fibres. Recent physiological evidence has demonstrated that some low-frequency cells in the anteroventral cochlear nucleus (AVCN) are sensitive to manipulations of the phase spectrum of complex sounds (Carney 1990b). These manipulations result in systematic changes in the spatiotemporal discharge patterns across groups of low-frequency AN fibres having different characteristic frequencies (CFS). One interpretation of these results is that these neurons in the AVCN receive convergent inputs from AN fibres with different CFS, and that the cells perform a coincidence detection or cross-correlation upon their inputs. This report presents a model that was developed to test this interpretation.


Neural organization and responses to complex stimuli in the dorsal cochlear nucleus. The dorsal division of the cochlear nucleus (DCN) is the most complex of its subdivisions in terms of both anatomical organization and physiological response types. Hypotheses about the functional role of the DCN in hearing are as yet primitive, in part because the organizational complexity of the DCN has made development of a comprehensive and predictive model of its input-output processing difficult. The responses of DCN cells to complex stimuli, especially filtered noise, are interesting because they demonstrate properties that cannot be predicted, without further assumptions, from responses to narrow band stimuli, such as tones. In this paper, we discuss the functional organization of the DCN, i.e. the morphological organization of synaptic connections within the nucleus and the nature of synaptic interactions between its cells. We then discuss the responses of DCN principal cells to filtered noise stimuli that model the spectral sound localization cues produced by the pinna. These data imply that the DCN plays a role in interpreting sound localization cues; supporting evidence for such a role is discussed.


Binaural masking and sensitivity to interaural delay in the inferior colliculus. The binaural masking level difference (BMLD) is a psychophysical effect whereby signals masked by a noise at one ear become unmasked by sounds reaching the other. BMLD effects are largest at low frequencies where they depend on signal phase, suggesting that part of the physiological mechanism responsible for the BMLD resides in cells that are sensitive to interaural time disparities. We have investigated a physiological basis for unmasking in the responses of delay-sensitive cells in the central nucleus of the inferior colliculus in anaesthetized guinea pigs. The masking effects of a binaurally presented noise, as a function of the masker delay, were quantified by measuring the number of discharges synchronized to the signal, and by measuring the masked threshold. The noise level for masking was lowest at the best delay for the noise. The mean magnitude of the unmasking across our neural population was similar to the human psychophysical BMLD under the same signal and masker conditions.


Philosophy and stimulus design for neuroethology of complex-sound processing. In research on the neural mechanisms for the processing of biologically important sounds such as species-specific sounds and sounds produced by prey and predators, it is necessary to study responses of central auditory neurons to biologically important sounds, information-bearing elements (IBEs) in them, and tone bursts. The tone bursts or constant-frequency (CF) components can be an IBE in many species of animals. Information-bearing parameters characterizing these sounds must be systematically varied, and tuning of neurons to individual parameters must be studied. The measurement of a tuning curve must be performed not only for excitatory responses, but also for inhibitory and facilitative responses, if any. The selectivity of a neuron to a particular type of sound must be tested for whether it is level-tolerant. Responses to complex sounds can probably be explained on the basis of those to IBEs and tone bursts, so that the use of the tone bursts, even though they are not IBEs, is as essential as that of the biologically important sounds.


Effect of ammonium ions on synaptic transmission in the mammalian central nervous system. It is not surprising that a compound with such unique properties as NH3/NH4+, should have a large variety of biochemical and neurological effects and to find itself implicated in many pathological conditions. Its undissociated (NH3) or dissociated (NH4+) forms, having different physicochemical properties, enter neurons and other cells through differing pathways. These two forms then change internal pH in opposite directions, and initiate a variety of regulatory processes that attempt to overcome these pH changes. In addition, ammonia has a central role in normal intermediary metabolism, and when present in excess, it can disturb reversible reactions in which it participates. The challenge in interpreting these various observations lies in the difficulty in assigning to them a role in the generation of symptoms seen in experimental and clinical hyperammonemias. In this review we have attempted to summarize information available on the effects of ammonium ions on synaptic transmission, a central process in nervous system function. Evidence has been presented to show that ammonium ions, in pathologically relevant concentrations, interfere with glutamatergic excitatory transmission, not by decreasing the release of glutamate, but by preventing its action on post-synaptic AMPA receptors. Furthermore, NH4+ depolarizes neurons to a variable degree, without consistently changing membrane resistance, probably by reducing [K+]i. A decrease in EK+ may also be responsible for decreasing the effectiveness of the outward chloride pump, thus explaining the well known inhibitory effect of NH4+ on the hyperpolarizing IPSP. There is a consensus of opinion that chronic hyperammonemia increases 5HT turnover and this may be responsible for altered sleep patterns seen in hepatic encephalopathy. There does not seem to be a consistent effect on catecholaminergic transmission in hyperammonemias. However, chronic hyperammonemia causes pathological changes in perineuronal astrocytes, which may lead to a reduced uptake of released glutamate and a decreased detoxification of ammonia by the brain. Chronic moderate increase in extracellular glutamate results in a down-regulation of NMDA receptors, while the decreased detoxification of ammonia makes the central nervous system more vulnerable to a sudden hyperammonemia, due, for instance, to an increased dietary intake of proteins or to gastrointestinal bleeding in patients with liver disease. Clearly, data summarized in this review represent only the beginning in the elucidation of the mechanism of ammonia neurotoxicity. It should help, we hope, to direct future investigations towards some of the questions that need to be answered.


Non-MHC-restricted T-cell interaction with B cells: role of the T-cell receptor. Non-specific antibody production usually accompanies the T-cell-regulated B-cell response. In this paper the mechanisms involved in non-MHC-restricted T-B-cell interaction were studied. As previously shown for NK cells, activated B cells induce IFN-gamma and TNF alpha production in non-MHC-restricted cytotoxic T lymphocytes (NrCTL). Using an in vitro model system, we demonstrate that direct cell-cell interactions are required to induce these cytokines in NrCTL. Receptor ligand systems involved are leucocyte function antigen-1/intercellular adhesion molecule-1 (LFA-1/ICAM-1)(CD11a, CD18/CD54), T11/LFA-3 (CD2/CD58), and the clonotypic T-cell receptor (TCR) structure NKTa of JT9/JT10 with its non-MHC-related target antigen TNKtar (4F2). Cytokine production can be induced by activating monoclonal antibodies against CD2R. Antibodies against the clonotypic TCR (NKTa) or CD3 had no cytokine-inducing effect on NrCTL cultured alone, but were able to retrieve the effect of blocking the target antigen on co-cultured B cells. We could further demonstrate that the inhibition of the TCR/target antigen interaction could be overcome by close cell-cell contact culture conditions. From these findings it is concluded that the role of the TCR in non-MHC-restricted cell-cell interaction is to facilitate LFA-1/ICAM-1-mediated effector target adhesion in a specific way rather than to mediate direct activating signals upon lymphokine production or cytotoxicity.


Control of Trypanosoma cruzi infection in mice deprived of T-cell help. The role of CD4+ T lymphocytes in the resistance of BALB/c mice to Trypanosoma cruzi was examined by in vivo depletion using monoclonal anti-CD4 antibodies (MoAbs). When the administration of MoAbs was initiated 2 days before, or 5 to 12 days after the infection (dpi) with 50 bloodstream-form trypomastigotes of the Tulahuén strain, mice showed an enhanced susceptibility to the parasite. Specific IgG, but not IgM responses, were inhibited in anti-CD4-treated and infected mice. However, when anti-CD4 treatment of mice was delayed until the 8th week of infection, neither a reactivation of the infection as determined by mortality or parasitaemia, nor a modulation of the titre of anti-T. cruzi IgG antibodies was detected. Furthermore, mice chronically infected with T. cruzi and deprived of CD4+ T cells resisted the challenge with 50,000 trypomastigotes (approximately 1000 LD50). Secondary antibody responses against parasite antigens were inhibited after in vitro depletion of CD4+ cells in chronically infected mice before boosting with T. cruzi antigens. However, recipients of CD4 or T-cell-depleted spleen cells from mice chronically infected with T. cruzi were protected when challenged with the parasite. The possibility that the parasite control is maintained by long-lived B cells capable of rapid differentiation into IgG-secreting plasma cells in the absence of T helper cells is discussed considering the present data.


Treatment of rheumatoid arthritis with single dose or weekly pulses of chimaeric anti-CD4 monoclonal antibody. The aetiology of rheumatoid arthritis is unknown but CD4+ T cells are known to be involved in its pathogenesis. Because of this, anti-CD4 monoclonal antibody has been used in open studies with clinical benefit in up to 60% of patients. We have used a chimaeric anti-CD4 monoclonal antibody (cM-T412, Centocor) in a randomized, double-blinded, placebo controlled trial as treatment for rheumatoid arthritis. Nine patients with active rheumatoid arthritis resistant to traditional disease-modifying drugs were recruited. Four received an intravenous 50 mg bolus of antibody, and three received 50 mg weekly for four consecutive weeks. Two patients received placebo. Despite a marked reduction (P less than 0.001) in peripheral blood CD4+ lymphocytes, there was no significant clinical improvement in any of these patients. The decrease in CD4+ lymphocyte number lasted one week after a single 50 mg dose of cM-T412 but was more prolonged in the patients who received four infusions. CD8+ T cells, CD16+ cytotoxic cells and CD14+ monocytes showed only a transient reduction. It may be concluded that the therapeutic efficacy of anti-CD4 therapy is not directly related to CD4+ T-cell lymphopenia.


Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Effective methods are needed to identify and isolate those genes that are differentially expressed in various cells or under altered conditions. This report describes a method to separate and clone individual messenger RNAs (mRNAs) by means of the polymerase chain reaction. The key element is to use a set of oligonucleotide primers, one being anchored to the polyadenylate tail of a subset of mRNAs, the other being short and arbitrary in sequence so that it anneals at different positions relative to the first primer. The mRNA subpopulations defined by these primer pairs were amplified after reverse transcription and resolved on a DNA sequencing gel. When multiple primer sets were used, reproducible patterns of amplified complementary DNA fragments were obtained that showed strong dependence on sequence specificity of either primer.


A point mutation of the alpha 2-adrenoceptor that blocks coupling to potassium but not calcium currents. The alpha 2A-adrenergic receptor (adrenoceptor) was stably expressed in AtT20 mouse pituitary tumor cells; adrenoceptor agonists inhibited adenylyl cyclase, inhibited voltage-dependent calcium currents, and increased inwardly rectifying potassium currents. An aspartic acid residue (Asp79) highly conserved among guanine nucleotide-binding protein (G protein)-coupled receptors was mutated to asparagine; in cells transfected with the mutant alpha 2-receptor, agonists inhibited adenylyl cyclase and calcium currents but did not increase potassium currents. Because distinct G proteins appear to couple adrenoceptors to potassium and calcium currents, the present findings suggest that the mutant alpha 2-adrenoceptor cannot achieve the conformation necessary to activate G proteins that mediate potassium channel activation.


Membrane properties of identified mesencephalic dopamine neurons in primary dissociated cell culture. Dopamine (DA)-containing neurons in primary dissociated cell cultures derived from the embryonic mouse mesencephalon (day E13) were studied by histochemical and electrophysiological techniques. DA neurons exhibited two distinct morphologies, fusiform and multipolar, tended to reside in groups and organize dendrites into common fascicles. While these neurons expressed the cell-surface marker acetylcholinesterase, the presence of this enzyme could not be used to identify DA neurons unequivocally, since it was also observed in nondopaminergic cells. Neurons were therefore identified as DA by their distinct morphology, and this identification was validated with a double-labeling procedure that entailed the intracellular deposition of a fluorescent dye (Lucifer yellow or ethidium bromide), followed by processing for tyrosine hydroxylase immunocytochemistry. DA neurons identified in this manner were observed to have resting membrane potentials between -50 and -75 mV, input resistances of 50-360 M omega, and membrane time constants of 4.1-14.1 msec. Forty-seven percent of these cells displayed spontaneous activity that was irregular in nature and often contained bursts (burst length was between two and six action potentials). The DA neurons displayed a variety of ionic conductances, including (1) a Na+ conductance (gNa) that underlies the action potential, (2) Ca2+ conductances (gCa) that mediate the nonsomatic low- and high-threshold spikes observed, and (3) at least three K+ conductances (gK). Voltage-clamp analysis revealed several distinct transmembrane ionic currents, including (1) a large, rapidly inactivating tetrodotoxin-sensitive inward Na+ current (INa), (2) a 4-aminopyridine-sensitive, transient early outward K+ current that required a conditioning hyperpolarization of the membrane to be activated by a subsequent depolarization (A-current, IA), (3) a slowly developing inward current that was seen only after a conditioning hyperpolarization of the membrane and that was dependent on the presence of external Ca2+ ions (ICa), and (4) a late-onset, noninactivating K+ current. Between 25% and 54% of the late-onset K+ current was Ca(2+)-dependent and was not affected by tetraethylammonium ions. This current was termed IAHP. The remaining current was not sensitive to changes in the extracellular Ca2+ concentration but was blocked by external tetraethylammonium. This current was termed IK. The direct pressure application of DA (1-200 microM) onto the soma dose-dependently hyperpolarized these neurons; this effect was potentiated by the presence of the catecholamine reuptake blocker cocaine hydrochloride (10-200 microM). Under voltage-clamp conditions, DA was observed to increase IK significantly and had little effect on IAHP.(ABSTRACT TRUNCATED AT 400 WORDS)


Induction of long-term potentiation in the basolateral amygdala does not depend on NMDA receptor activation. Long-term potentiation (LTP) can be induced in the lateral and basolateral amygdala by stimulating synaptic afferents in the external capsule (EC). We examined the sensitivity of amygdaloid LTP to the NMDA receptor antagonist 2-amino-5-phosphonopentanoate (AP5), which is known to block LTP induction in the Schaffer collateral/CA1 synapses in the hippocampus. While relatively high concentrations (100 microM) of DL-AP5 were effective in preventing LTP induction in the lateral and basolateral amygdala in vitro, the same concentrations also significantly depressed synaptic responses to low-frequency stimulation. Furthermore, at 50 microM, a concentration sufficient to block both synaptic responses mediated by NMDA receptors and LTP induction in the hippocampus and neocortex, AP5 did not affect the probability of inducing LTP in the amygdala. Application of 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), which blocks non-NMDA excitatory amino acid receptors, reduced the monosynaptic response to EC stimulation by 85%. The remaining CNQX-insensitive response did not appear to be mediated by NMDA-type receptors, since it was not reduced by 50 or 100 microM AP5, and showed none of the voltage sensitivity characteristic of NMDA responses. These data suggest that while the induction of LTP in the amygdala produced by EC stimulation is blocked by high doses of AP5, plasticity at these synapses probably does not require activation of NMDA receptors.


Microiontophoretic studies of the effects of D-1 and D-2 receptor agonists on type I caudate nucleus neurons: lack of synergistic interaction. Several lines of evidence have suggested there may be a physiologically relevant form of synergistic interaction between D-1 and D-2 dopamine (DA) receptors located on postsynaptic neurons in the forebrain that receive a dopaminergic innervation. Because of the theoretical importance of such an interaction with respect to understanding the normal physiology of dopaminergic systems, we evaluated effects of D-1 and D-2 selective agonists, applied microiontophoretically, on the spontaneous electrical activity of a single, identifiable subpopulation of neurons within the caudate nucleus, the type I striatal neuron, in locally anesthetized, gallamine-paralyzed rats. It was observed that the D-2 receptor agonist quinpirole (QUIN) produced biphasic effects on cell firing rate. Low ejection currents significantly increased firing rate, while higher currents produced an inhibition. Similar effects were observed for the D-1 agonists SKF 38393; however, the overall excitations observed at low ejection currents were far less than those observed for QUIN. When these two agonists were applied concurrently, a simple additive effect (but not synergism) was always observed. The acute reduction of striatal levels of DA, by as much as 84% (with pretreatment with alpha-methyl-p-tyrosine, AMPT), did not alter the responsiveness of type I striatal neurons to the DA receptor agonists applied alone or in combination. These observed effects were not altered either by chloral hydrate anesthesia (in which glutamate-driven activity was studied) or by a more severe depletion of striatal DA levels (98% depletion produced by combined pretreatment with AMPT and reserpine).


Differential modulation by dopamine of responses evoked by excitatory amino acids in human cortex. The responses of human neocortical neurons to iontophoretic application of excitatory amino acids and their modulation by dopamine (DA) were studied in vitro. Brain slices were obtained from children undergoing surgery for intractable epilepsy. Application of N-methyl-D-aspartate (NMDA) to the slices induced slow depolarizations accompanied by decreased input conductances and sustained action potentials in cortical neurons. Glutamate produced rapid depolarizations and firing with few changes in input conductances. Quisqualate also induced depolarization and firing, but input conductances increased during the rising phase of the membrane depolarization. Iontophoretic application of DA alone produced no change in membrane potential or input conductance. However, when DA was applied in conjunction with the excitatory amino acids, it produced contrasting effects. With either bath application of DA or when iontophoresis of DA preceded application of NMDA, the amplitude of the membrane depolarizations and the number of action potentials were increased, whereas the latency of these responses decreased. In contrast, DA decreased the amplitude of the depolarizations and the number of action potentials evoked by glutamate or quisqualate. The fact that DA affects responses to NMDA and glutamate or quisqualate in opposite directions is of considerable importance to the understanding of cellular mechanisms of neuromodulation and the role of DA in cognitive processing and in epilepsy.


Modification of atracurium or vecuronium blockade and their reversal by succinylcholine in the cat. The interaction between succinylcholine (SCC) and non-depolarizers, atracurium or vecuronium was investigated in 36 cats of either sex using the sciatic nerve-anterior tibialis muscle preparation. Additionally, the relation of SCC to pseudocholinesterase activity was examined. The duration of action of vecuronium (6.5 +/- 1.3 to 7.3 +/- 2.2 minutes) in cats pretreated with SCC was greater than those (2.0 +/- 0.6 minutes) in non-pretreated cats. However, SCC had no influence on the duration of atracurium. The serum pseudocholinesterase activity was decreased after the injection of atracurium or neostigmine in contrast to vecuronium. The authors conclude that the prior administration of SCC prolongs the duration of vecuronium but not that of atracurium, and pseudocholinesterase activity is not related to the prolonging effect of SCC.


The origin of brainstem noradrenergic and serotonergic projections to the spinal cord dorsal horn in the rat. Although it has been proposed that the locus coeruleus is the predominant, if not exclusive, brainstem origin of the noradrenergic innervation of the spinal dorsal horn, pharmacological studies argue otherwise. In this study we made localized injections of the retrograde tracer wheatgerm agglutinin conjugated to apo-horseradish peroxidase gold (WGA:apoHRP-Au), in conjunction with immunocytochemical labeling for tyrosine hydroxylase (TH) or serotonin (5-HT), to identify the brainstem source of the noradrenaline (NA) and 5-HT innervation of the dorsal horn of the rat. Our studies were concentrated in the C5 spinal segment. The pattern of labeling was only studied in animals in which the tracer injection was restricted to the dorsal horn. In these rats, TH-immunoreactive neurons in widespread regions of the brainstem, including the locus coeruleus, subcoeruleus, A5, and A7 cell groups, were found to project to the dorsal horn. In terms of absolute numbers of double-labeled cells, no one noradrenergic cell group predominated. As expected, dorsal-horn-projecting 5-HT-immunoreactive neurons were found within the 5-HT populations of the rostroventromedial medulla and caudal pons, including the nucleus raphe magnus, nucleus paragigantocellularis (PGi), and ventral portions of the nucleus gigantocellularis (Gi). The majority of retrogradely labeled 5-HT-immunoreactive cells were, however, located off the midline, in the ipsilateral PGi and ventral Gi. Finally, a large number of retrogradely labeled, non-5-HT cells were found intermingled among the 5-HT cells of this region. Our results provide evidence that the noradrenergic regulation of nociceptive transmission at the spinal cord level arises from direct spinal projections of several brainstem noradrenergic cell groups.


Annulate lamellae in a large cell lung carcinoma cell line with high expression of tyrosine kinase receptor and proto-oncogenes. The morphology, karyotype, in vitro growth properties, and expression of tyrosine kinase receptors and proto-oncogenes are reported for a newly established large cell undifferentiated lung carcinoma cell line (RVH-6849). The results were analyzed concomitantly with those for two well-established cell lines from an adenocarcinoma of the lung (A549) and a squamous cell carcinoma (A431). All three cell lines demonstrated common ultrastructural features of epithelial cells, but only RVH-6849 had frequent aggregates of centrioles and annulate lamellae (AL) and was polyploid, having five to seven copies of chromosome 7 by karyotype analysis. All three cell lines expressed transforming growth factor alpha (TGF-alpha), epidermal growth factor receptor (EGFR), c-erb B-2, and c-met genes. RVH-6849 cells, however, expressed the most messenger RNA (mRNA) for TGF-alpha, c-erb B-2, and c-met. Only EGFR mRNA was expressed more in the other two cell lines, especially in A431 cells. AL represent an exaggerated form of the nuclear membrane-pore complex that is found in actively proliferating cells such as germ and some neoplastic cells. AL are suspected to be involved in the deposition or processing of mRNA: The enhanced coexpression of AL and mRNAs of three tyrosine kinase-containing receptors in RVH-6849 cells may represent such a relationship.


Role of chemotherapy in the future treatment of ovarian cancer. Platinum-based chemotherapy has led to an improvement in complete response rates and duration of median remission, but has only given a modest improvement in overall survival in patients with advanced ovarian cancer. Chemotherapy will in the future focus upon: (1) improving the complete remission rate with new induction regimens; (2) identifying strategies capable of converting partial remission into complete remission; (3) preventing or delaying recurrences in patients who do achieve a complete remission; (4) identifying mechanisms of antineoplastic drug resistance and pharmacologic techniques capable of reversing drug resistance. Among the treatment approaches being utilized are high-dose chemotherapy with autologous bone marrow transplantation, development of new chemotherapeutic regimens which include Taxol and hexamethylmelamine, and intraperitoneal chemotherapy. In addition, our understanding of the mechanisms of antineoplastic drug resistance has led to the development of novel therapeutic approaches. It has been demonstrated that resistance to platinum and alkylating agents is associated with both increased concentrations of cellular glutathione (GSH) as well as an increased capacity of tumor cells to repair damage to DNA. Inhibition of GSH biosynthesis with buthionine sulfoximine (BSO), a synthetic inhibitor of the enzyme gamma glutamyl cysteine synthetase, has led to the potentiation of alkylating agent activity in vitro and in vivo. A phase I trial of BSO plus melphalan is currently in progress and a trial of BSO plus carboplatin is planned. Inhibition of the DNA repair process with aphidicolin potentiates the cytotoxicity of cisplatin in drug-resistant tumor cells. Clinical trials of aphidicolin plus cisplatin await the completion of ongoing phase I trials of aphidicolin.


[Clinicopharmacology of Arduan].

[Clinicopharmacology of Arduan]. The paper presents clinicopharmacological investigations of Arduan carried out in Hungary and in other countries. The aim of the research work is to find an ideal muscle relaxant without side-effects, which are mainly cardiovascular. Pipecuronium bromide (Arduan) is a compound with steroid structure; it has a long duration of action and, most importantly, it does not affect circulation. The properties of pipecuronium bromide will be discussed and compared to those of pancuronium and vecuronium bromide.


Myocardial potassium balance associated with regional ischaemia in the pig: effects of beta-adrenoceptor blockade, duration of ischaemia and preceding ischaemic periods. The effects of beta-adrenoceptor blockade, duration of ischaemia and preceding ischaemic periods on ischaemia-induced changes in myocardial K+ balance were studied in 12 open-chest pigs. Coronary venous blood was directed through a shunt from the coronary sinus to the right atrium. Continuous recordings of arterial, shunt blood [K+] and shunt flow enabled us to compute myocardial K+ balance during and after consecutive 2-, 2-, 5-, 10- and 2-min periods of regional ischaemia separated by 30 min of reperfusion. beta-adrenoceptor blockade (propranolol 1 mg kg-1 i.v.) given between the first and second ischaemic period did not alter the effects of 2 min ischaemia on myocardial K+ balance. Total K+ losses induced by 2, 5 and 10 min of ischaemia were 67.1 (40.6-93.3), 106.7 (69.4-176.8) and 192.2 (117.7-332.6) mumol 100 g-1, respectively. Thus, the plateau observed in extracellular [K+] between 2 and 10 min of regional ischaemia could, at least partly, be explained by continuous drainage of K+ from ischaemic myocardium into the surrounding normally perfused tissue. The total K+ loss induced by the second and last 2-min ischaemic period were 67.1 (40.6-93.3) and 35.6 (23.1-53.6) mumol 100 g-1 (P less than 0.001), respectively. This reduction shows that ion homeostasis during ischaemia was greatly changed in myocardium which had been 'preconditioned' and 'stunned' by 5 plus 10 min of ischaemia. Total amount, maximal rate and duration of post-ischaemic K+ reuptake increased with the duration of the preceding ischaemia. Moreover, K+ re-uptake after 2 min of ischaemia and the number of sarcolemmal Na/K pumps ([3H]ouabain binding), were normal in stunned myocardium. From these observations we conclude that progressive stimulation of the Na/K-pump occurred when ischaemia was prolonged from 2 to 10 min, and that Na/K-pump function was preserved in stunned myocardium.


Establishment and characterization of an etoposide-resistant human small cell lung cancer cell line. An etoposide-resistant subline, SBC-3/ETP, from a human small cell lung cancer cell line, SBC-3, was developed by continuous exposure to increasing concentrations of etoposide in culture. The SBC-3/ETP was 52.1-fold more resistant to etoposide than the parent cell line. The SBC-3/ETP was highly cross-resistant to teniposide, adriamycin, vinca alkaloids, 4-hydroperoxycyclophosphamide, CPT-11 and mitomycin C, and marginally cross-resistant to cisplatin, while the subline showed a collateral sensitivity to bleomycin. Topoisomerase I activity in the SBC-3/ETP was reduced to an extent of one half and topoisomerase II activity to an extent of one eighth in comparison with those of the SBC-3. Intracellular accumulation of [3H]-etoposide in the SBC-3/ETP was significantly lower in comparison to the SBC-3. An overexpression of MDR1 mRNA, and the presence of its product, P-glycoprotein, were detected in the SBC-3/ETP by Northern blotting and flowcytometry using a monoclonal antibody of the protein, MRK16. These results indicate that a decreased activity of topoisomerase II is the major factor for the development of etoposide resistance, and that an overexpression of the MDR1 gene is responsible, in part, for the development of resistance to the drug and some structurally unrelated compounds such as adriamycin and vinca alkaloids.


Hormonal profiles in Italian late-onset adrenal hyperplasia correlate with HLA class III polymorphisms. To investigate the genetic polymorphisms of the HLA region in late-onset adrenal hyperplasia, 13 Italian patients affected by the disease were analyzed for: (1) HLA-A and -B typing; (2) restriction fragment length polymorphism (RFLP) of DR beta, DQ beta, DQ alpha, 21-hydroxylase A and B genes; (3) fourth complement fraction loci A and B (C4A and C4B), second complement fraction (C2) and properdin B factor (Bf) complement typing; (4) hormonal characteristics associated with some HLA haplotypes. HLA alleles B14 and DR beta 1 were found to be significantly more frequent in patients with respect to controls (relative risk: 8.7 and 7.2, p less than 0.001 and p less than 0.0001, respectively). Also C4B*2, 1 duplication was more frequent in patients than in normal subjects (23% vs. 1.5%, p less than 0.0001). Moreover, patients carrying a duplicated C4B (as well as those having the B14 antigen) showed higher 17-hydroxyprogesterone levels after ACTH stimulation. RFLP analysis of 21-hydroxylase genes with a specific probe revealed a duplication of 21-hydroxylase A gene in 40% of patients. All these individuals carried the C4A*2 B*2,1 phenotype and 75% of them displayed a clearly recognizable duplication at the C4B locus. These data support the hypothesis that in late-onset adrenal hyperplasia the 21-hydroxylase A pseudogene, even if inactive, may play a negative role in the regulation of 21-hydroxylase biosynthesis. Furthermore, we suggest analyzing class III phenotypes to screen the enzymatic defect.


[Effect of 8-hydroxycarteolol in normal human eyes]. The authors studied the ocular hypotensive effect of 8-hydroxycarteolol, a main metabolite of carteolol in normal human eyes, after topical administration in a double masked randomized study. After instillation of 0.01%, 0.1%, 0.5% and 1% ophthalmic solutions of 8-hydroxycarteolol, statistically significant reduction of intraocular pressure was observed 8 and 24 hours after administration (p less than 0.05). It's maximum reduction of intraocular pressure, 1.6 +/- 0.5 mmHg was produced 24 hours after treatment by 0.1% solution. However, diurnal variation of IOP did not show a statistically significant difference.


Phorbol ester-induced hairy cell features on chronic lymphocytic leukemia cells in vitro. Leukemic cells from eight patients with chronic lymphocytic leukemia were isolated and cultured in the continuous presence of 12-O-tetradecanoylphorbol 13-acetate (TPA) at a concentration of 1.6 x 10(-9) M for 4-10 days. Aliquots of cells were then analyzed at intervals of 24-72 hr for changes in morphology, acid phosphatase staining (AP), and expression of two hairy cell-associated surface antigens, HCL1 (CD22, Leu 14) and HCL3 (CD11c, Leu M5). All cases studied showed typical B-CLL phenotype, and only a small proportion of cells expressed CD22 and CD11c (mean 7% and 4.9%, respectively). TPA treatment induced the coexpression of CD22 (mean 49%) and CD11c (mean 48%) and tartrate-resistant acid phosphatase in seven of eight cases. Morphologically, cells in TPA cultures expressed hairy cell features that were evident in light and electron microscopic studies. Collectively these changes indicate that TPA can induce hairy cell features on CLL cells in vitro, suggesting the later maturational stage of HCL compared with CLL.


Tardive dyskinesia in elderly psychiatric patients: a 5-year study. The authors investigated the prevalence of tardive dyskinesia among elderly psychiatric patients who had never received neuroleptic medication before their first hospitalization. The study was performed in the geriatric psychiatry unit of a university-affiliated hospital in Canada and involved all first-admission patients admitted from September 1984 through August 1989 who had never taken neuroleptic drugs. In September and October 1989, the patients who were available for follow-up were examined and given ratings on the Abnormal Involuntary Movement Scale to establish the presence or absence of tardive dyskinesia. The patients' records were reviewed for information on age, diagnosis, duration of hospitalization, neuroleptic treatment received after admission, anticholinergic drugs received, and drug-free periods. Of the 162 patients who were available and whose data were analyzed, a total of 99 had been treated with neuroleptics, and 35 (35.4%) of these were found to have tardive dyskinesia. Two of the 35 also had tardive dystonia. Significantly more patients with major depression than patients with primary degenerative dementia or delusional psychosis had tardive dyskinesia. This study confirms the higher vulnerability of elderly psychiatric patients treated with neuroleptics to the development of tardive dyskinesia. The authors stress that caution is especially necessary when neuroleptics are prescribed for older patients with major affective disorders.


Vector density gradients and the epidemiology of urban malaria in Dakar, Senegal. The dispersion of anopheline mosquitoes from their breeding places and its impact on malaria epidemiology has been investigated in Dakar, Senegal, where malaria is hypoendemic and almost exclusively transmitted by Anopheles arabiensis. Pyrethrum spray collections were carried out along a 910-meter area starting from a district bordering on a permanent marsh and continuing into the center of the city. According to the distance from the marsh, vector density (the number of An. arabiensis per 100 rooms) at 0-160, 160-285, 285-410, 410-535, 535-660, 660-785, and 785-910 meters was 84, 40, 5, 2, 2, 0.4, and 0, respectively, during the dry season, and 414, 229, 110, 84, 99, 69, and 21, respectively, during the rainy season. The proportion of 8-11-year-old children with negative immunofluorescent antibody test results for Plasmodium falciparum was 17%, 28%, 44%, 54%, 50%, 63%, and 73%, respectively, in these different sections. Malaria prevalence in the community was maximum in the area bordering on the marsh where it ranged from 1% to 15% (average 6%) according to age and season of the year. These findings show the epidemiologic importance of vector density gradients in Dakar. The implications for malaria control in urban areas are discussed.


Molecular characterization of Mexican stocks of Trypanosoma cruzi using total DNA. Seventeen Mexican Trypanosoma cruzi stocks and five South American reference strains were analyzed by Hind III restriction fragment length polymorphisms associated with ribosomal RNA gene spacers and by HinfI digestion patterns of total DNA. Our findings demonstrate the occurrence of genetic heterogeneity within these stocks. Hierarchic and non-hierarchic clustering of these molecular characters allowed the formation of groups that correlate with the geographic origin of the stocks. The HinfI digestion pattern permitted the identification of DNA fragments from the kinetoplast maxi-circles, and therefore represents a simple and convenient method for conducting epidemiologic surveys in laboratories in developing countries.