Combining markers of nephrotoxicity and hepatotoxicity for improved monitoring and detection of chronic alcohol abuse.

BACKGROUND: Most of the commonly used markers of chronic alcohol abuse reflect alcohol hepatotoxicity; however, such abuse is deleterious to the kidneys as well. Combined use of serum markers of liver origin and urinary markers of kidney origin may be of diagnostic advantage. METHODS: The study was performed in 73 male alcoholics undergoing detoxification and 36 male alcoholics who had maintained abstinence for > or =6 weeks. Factor analysis, discriminant analysis and receiver operating characteristic (ROC) analysis were used to assess the discriminative power of two urinary markers of alcohol nephrotoxicity, namely beta-N-acetylhexosaminidase (Hex, EC 3.2.1.52) and alanine aminopeptidase (EC 3.4.11.2), and of three serum markers of alcohol hepatotoxicity, namely aspartate aminotransferase (EC 2.6.1.1), alanine aminotransferase (EC 2.6.1.2) and gamma-glutamyltransferase (GGT, EC 2.3.2.2), and of their quantitative combinations. RESULTS: The discriminative power of the urinary markers matched that of the serum markers. Hex and GGT appeared to be the best for discriminating the study groups. Their combination given by the equation G&H=0.62 x ln(GGT)+0.72 x ln(Hex) showed excellent discriminative ability (ROC area under the curve 0.92) that was significantly higher than that of any single marker in this report, indicating superior diagnostic accuracy of the compound marker. CONCLUSIONS: Kidney-derived urinary markers, particularly Hex, can complement or replace, if necessary, serum markers of chronic alcohol abuse that relate to alcohol hepatotoxicity. The compound marker proposed seems a promising tool for the monitoring and perhaps detection of chronic alcohol abuse and warrants further studies.

Anxiolytic-like effects of mGlu1 and mGlu5 receptor antagonists in rats.

The purpose of the present study was to compare anxiolytic activity of the metabotropic glutamate receptor 1 (mGlu) antagonist, EMQMCM ((3-ethyl-2-methyl-quinolin-6-yl)-(4-methoxy-cyclohexyl)-methanone methanesulfonate) and the mGlu5 receptor antagonist MTEP ([(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine) and MPEP (2-methyl-6-(phenylethynyl)pyridine) in animal models of anxiety. In the elevated plus maze, diazepam (1 mg/kg), but not the mGlu1 or mGlu5 receptor antagonists induced anxiolytic-like effects. Meanwhile, MTEP (2.5 and 5 mg/kg), EMQMCM (5 mg/kg), and diazepam (2 mg/kg) all significantly inhibited fear potentiated startle. In the contextual fear conditioning test, MTEP (1.25 and 2.5 but not 5 mg/kg) and EMQMCM (0.6 to 5 mg/kg) attenuated freezing responding. In the Geller-Seifter conflict test, MPEP (1 and 3 mg/kg), MTEP (3 mg/kg), chlordiazepoxide (10 and 20 mg/kg) and midazolam (1 mg/kg) all facilitated punished responding, while ECMQCM failed to produce any significant effects up to 3 mg/kg dose. To summarise, the present data further support a significant anxiolytic potential of group I mGlu receptor antagonists, while suggesting the effects of mGlu1 receptor antagonists may depend on the experimental procedure and may be qualitatively different from those of mGlu5 receptor antagonists.

The effect of fluoxetine in a model of chemically induced seizures--behavioral and immunocytochemical study.

The aim of this study was to examine the effects of acute fluoxetine treatment on pentylenetetrazol-induced convulsions in order to shape a model of seizures associated with treatment with antidepressants in rats. Moreover, the putative role of the hippocampal formation in this respect was investigated with the help of c-fos immuncytochemistry to mark local neuronal activity. It was found that fluoxetine (10.0 mg/kg, i.p.) enhanced the proconvulsive effect of pentylenetetrazol (50.0 mg/kg, i.p.), and simultaneously inhibited pentylenetetrazol-stimulated c-Fos expression in some areas of the hippocampus. Fluoxetine pretreatment did not alter pentylenetetrazol brain concentration indicating that this phenomenon was not related to the pharmacokinetic interaction. It is suggested that inhibition by fluoxetine of some neuronal populations contributing to the local feedback mechanism controlling excessive epileptiform discharges within the hippocampus might lead to an increase in epileptic activity. The reported in the present paper fluoxetine versus pentylenetetrazol interaction may, therefore, serve as a model of seizures associated with treatment with antidepressants.

Neophobia and cortical and subcortical binding of the dopamine D2 receptor antagonist [3H]-raclopride.

The potential role of dopamine system in response to novelty was analysed using the selective dopamine D2 receptor antagonist, raclopride, in behavioral and biochemical assays, in rats (the open field test, and specific binding of [3H]-raclopride, within different brain structures measured with autoradiography). It was found that raclopride at a low dose (50 microg/kg, IP) caused anxiolytic-like effect (increased the anti-thigmotactic index), whereas at a higher dose (500 microg/kg, IP) produced general inhibitory influence, and decreased the anti-thigmotactic index. Analysis of the behavioral and biochemical results of the experiment revealed a significant negative correlation between the ligand binding in the substantia nigra pars reticulata (SNR), and the number of entries into the central sector of the open field (r=-0.48, p<0.05), as well as the positive correlation between time spent in the central sector of the open field and [3H]-raclopride binding within nucleus accumbens septi (r=0.57, p<0.05). Factor analysis revealed a Factor 1 (eigenvalue=3.361) grouping parameters of central entries into the open field and [3H]-raclopride binding in the SNR (factor loadings are 0.814 and 0.703 respectively), indicating that both phenomena are under control of a similar central process. The above data are discussed in relation to the structure dependent dopamine D2 receptor mechanisms in a rat response to novelty.

Age-dependent effects of 5,7-dihydroxytryptamine on serotonin transporter in different brain areas in the rat.

In the present study, we investigated the [(3)H]citalopram binding using a quantitative autoradiography following intracerebroventricular injection of 5,7-dihydroxytryptamine (5,7-DHT) in neonatal and adult male Wistar rats. One group of animals was injected with 5,7-DHT at 3 days after birth while the second group received the neurotoxin at 3 months after birth. Control group was injected with saline. Afterwards, all rats were examined at 4(th) months after birth to determine the serotonin (5-HT) and catecholamines concentrations using the liquid chromatography with electrochemical detection HPLC system and distribution and density of [(3)H]citalopram binding sites in the brain using the quantitative autoradiography. A marked depletion of brain 5-HT was observed in rats lesioned either in postnatal or adult period of life. Rats lesioned in their adult period of life showed dramatic reduction of 5-HT transporter in all investigated brain areas (i.e.the frontal cortex, entorhinal cortex, hippocampus, caudate-putamen, nucleus accumbens and ventral tegmental area). On the other hand, administration of 5,7-DHT to newborn rats failed to reduce 5-HT transporter sites in the ventral tegmental area, and produced only slight or moderate reduction in the nucleus accumbens. Thus, it appears that the mesolimbic ventral tegmental area-nucleus accumbens systems are relatively more resistant to 5,7-DHT neurotoxicity in the early postnatal period.

Sensitivity to pain and c-Fos expression in brain structures in rats.

The induction of c-Fos protein--a product of the c-fos gene, a marker of changes in neuronal activity, was studied in brain structures of animals differing in their sensitivity to the acute painful stimulation, a foot-shock (MS--more sensitive rats; LS--less sensitive rats, according to the arbitrary criterion in the flinch-jump pretest). After the pretest the animals were dived into the control group, exposed on retest 10 days later to the testing cage only (C1 group), and aversively stimulated animals (MS and LS groups, given five mild footshocks 1.5 h before immunocytochemical part of the experiment). Additional control group of naive, intact animals, was studied in parallel (C group). It was shown that animals subjected to the flinch-jump test retained a strong emotional reaction on re-exposure to the shock cage on retest (a conditioned fear) 10 days later, as revealed by the widespread expression of c-Fos protein in the examined brain structures, as compared with the control, naive rats not exposed to the testing cage. In the lateral habenular nucleus (LHAB) a similar effect has been found in the control animals re-exposed to the testing cage only (C1 group), and in the MS group, suggesting that this brain area participates predominantly in processing of emotional-cognitive component of a painful stimulation. In the periaqueductal gray and basolateral nucleus of amygdala the most pronounced, but significantly higher in comparison with C group only, expression of c-Fos was detected in MS rats. Interestingly, a strong and uniform enhancement of c-Fos expression appeared in all other brain structures examined, including cortical areas, indicating their sensitivity to non-direct (conditioned) aversive stimuli. The only significant difference in c-Fos expression between LS and MS rats found in LHAB points to this brain structure as selectively engaged in processing of the emotional-cognitive component of a painful stimulation. The reactivity of LHAB may be responsible for the genetically determined differences in sensitivity to pain.

Hippocampal mGluR1 and consolidation of contextual fear conditioning.

The effects of post-training intra-hippocampal injections of group I mGluR agonists and antagonists, were examined in the contextual fear test, in rats. It was found that (S)-3,5-dihydroxyphenylglycine (DHPG) (a mGluR1-5 agonist) decreased, and (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA) (a mGluR1 antagonist) increased fear conditioning (a freezing reaction), examined 24h after conditioning session. (RS)-2-Chloro-5-hydroxyphenylglycine (CHPG) (a mGluR5 agonist), and 2-methyl-6-(phenylethynyl)pyridine hydrochloride (MPEP) (a mGluR5 antagonist) did not cause any effect. In the immunocytochemical study, the post-conditioning administration of AIDA decreased the c-Fos induction in the dentate gyrus and CA1 layer of the hippocampus proper, 2h after exposure of animals to the aversive context, and 24h after conditioning session. It is suggested that overactivation of glutamatergic transmission in the critical for memory trace formation structure and period of time, may result in an attenuation of memory consolidation. On the other hand, reduction of an exaggerated glutamatergic tone can facilitate learning and memory processes. The immunocytochemical study and factor analysis of experimental data revealed that hippocampal mGlu1 receptors significantly influence the memory consolidation in a way dependant on the level of glutamatergic activity. Furthermore, they indicate that changes of glutamatergic activity within brain limbic structures can affect the threshold for the induction of the long-term neuronal plasticity, involved in some forms of learning and memory.

The role of neurosteroids in the anxiolytic,antidepressive- and anticonvulsive effects of selective serotonin reuptake inhibitors.

The introduction of SSRIs to the clinic was a turning point in the treatment of depression and related mental disorders. Nowadays, it is becoming more and more evident that SSRIs are equally effective in anxiety states and some types of epilepsy. However, the mechanism of their central action is not fully understood. They not only block the serotonin transporter, but also bind to different types of monoaminergic receptors (serotonergic, dopaminergic) and interact with the synthesis of neurosteroids, in this way modifying the excitability of the central nervous system. The aim of this review is to update information on the preclinical and clinical effects of SSRI, with special emphasis put on the probable role of neurosteroids and the GABAA receptor complex in the mechanism of their action.

[Neurodevelopmental theories of schizophrenia--preclinical studies]. / Neurorozwojowe koncepcje schizofrenii--badania przedkliniczne.

Schizophrenia is a complex disorder of unknown origin, characterised by abnormalities in the realms of perception, thinking and the experience of emotions that onset is restricted to young adulthood. Many techniques that range from neuropathology to neuroimaging identified subtle brain abnormalities particularly in frontal, temporal cortex, hippocampus, basal ganglia and cerebellum. Neurodevelopmental models of schizophrenia test hypotheses that this disease is caused by a defect in cerebral development which results in altered neural connectivity, brain neurochemistry and aberrant behaviour observed in adult life. Recent evidence indicates that neonatal hippocampal damage may affect prefrontal neuronal integrity. The developmental lesion model appears to have predictive validity because treatment with antipsychotic drugs normalises some abnormal behaviour changes. Therefore it will be a useful paradigm in the work on new therapies and in providing new insights about pathophysiology and etiology of schizophrenia.

Rat behavior in two models of anxiety and brain [3H]muscimol binding: pharmacological, correlation, and multifactor analysis.

The contribution of GABAergic mechanisms to rat emotional behavior in two animal models of anxiety (open field test of neophobia and aversively conditioned freezing reaction), was confirmed by pharmacological analysis, using anxiolytic (midazolam) and anxiogenic (picrotoxin) compounds. Both substances are known to modulate GABA(A) receptors' activity in a positive or negative manner, respectively. It seemed, therefore, worthwhile to check whether the behavioral parameters measured in these animal models of anxiety correlate with [3H]muscimol binding (a highly selective GABA(A) receptor ligand) in different brain structures of nai;ve rats, with a view to establish the role of genetically determined expression of local GABA(A) receptors in the organization of rat emotional and motor behavior. Correlation analysis revealed no links between individually determined expression of GABA(A) receptors (quantitative receptor autoradiography) in the brain structures, and the emotional behavior of nai;ve, drug-free animals, in both tests. Factor analysis confirmed that animal behavior in both tests was under control of different central processes. Moreover, none of the behavioral and ligand binding parameters loaded on the same factor, confirming the negative results of the correlation study. The present results indicate that the origin of emotions is a complex phenomenon, probably involving the interaction between GABA-ergic innervation of many brain structures.

Lack of changes in cortical [3H]-muscimol binding in rats sensitized to nicotine-induced enhancement of dopamine metabolism.

It was proposed that chronic nicotine treatment may induce adaptive changes in GABAA receptors, thus leading to the attenuation of a GABAergic inhibition of dopaminergic neurons. This putative mechanism might underlie the sensitization to nicotine-induced increase in locomotor activity and dopamine metabolism; i.e. phenomena highly significant to the dependence-producing effects of this psychostimulant. To test this hypothesis, in the present study we have analyzed the influence of acute and repeated treatment of rats with nicotine on the binding of a highly selective and competitive GABAA receptor agonist, [3H]-muscimol. The binding was investigated by autoradiography in different brain cortical structures. It was found that nicotine given at the dose stimulating locomotor activity (0.6 mg/kg, sc), markedly increased striatal HVA concentration in the group of animals chronically pretreated (for 6 days) with this psychostimulant. Neither acute nor repeated nicotine administration changed in a significant way the [3H]-muscimol binding to brain cortical structures. Thus, the hypothesis about the role of adaptive changes in GABAA receptors in the enhancement of the biochemical and behavioral effects of nicotine was not confirmed.

Effects of pentylenetetrazol-induced kindling of seizures on rat emotional behavior and brain monoaminergic systems.

The influence of pentylenetetrazol (PTZ)-induced kindling of seizures on the rat emotional behavior, the brain monoamine turnover rate measured in vitro, and correlation between behavioral and biochemical parameters, were examined in rats. The repeated administration of PTZ (35 mg/kg, ip) evoked kindled seizures in rats (Stage 4 or 5 of clonic-tonic convulsions-maximum). PTZ kindling caused selective changes in the rat emotional behavior, present in some models of anxiety only (a decreased freezing time in the conditioned freezing test and a decreased spontaneous and aversively conditioned ultrasonic vocalization). Simultaneously, PTZ kindling decreased the concentration of homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) in the prefrontal cortex, decreased the DA (HVA/DA ratio) turnover rate in the striatum, and inhibited the serotonin (5-HT) metabolism (5-HIAA/5-HT ratio) in the hippocampus and the prefrontal cortex. Correlations between dopamine (DA) or 5-HT regional metabolic rates in brain structures and animal behavior were either abolished or reversed in PTZ-kindled animals. It is concluded that both DA and 5-HT systems contribute to the emotional effects of PTZ-induced kindling of seizures. The hypothesis is put forward that PTZ kindling-induced inhibition of the serotonergic innervation may lead to the compensatory increase in 5-HT(1A) receptors in the dentate gyrus of the hippocampus, thus evoking the anxiolytic-like changes in animal behavior.