[The problems of expert evaluation of the results of forensic chemical investigations of clozapine in the biological material].

The objective of the present review was to analyse the problems of expert evaluation of the results of forensic chemical investigations of clozapine in the biological material. Such an analysis is needed because many topical aspects of the quantitative evaluation of toxic clozapine concentrations remain unclear. The treatment with clozapine is associated with its accumulation in blood in concentrations up to 2 mg/l in the absence of any toxic effect allegedly due to the development of tolerability of this agent. In the tolerant patients the ratio of the main clozapine metabolite, norclozapine, to clozapine itself in the serum amounts to 0.6-0.9. This value falls down to 0.3-0.4 in case of acute intoxication. In the case of identification of other pharmaceutical products narcotic drugs together with clozapine their influence on the activity of enzymes responsible for clozapine biotransformation should be taken into consideration. The concomitant intake of clozapine and alcohol may be dangerous for the clozapine-intolerant subjects. It is concluded that the above observations must be borne in mind in the assessment of the results of forensic chemical analysis.

[Forensic medical diagnostics of acute and chronic alcohol intoxication].

The principal characteristics and results of experimental studies on the problem of acute and chronic alcohol intoxication are presented. The mechanisms of toxic action of ethanol and acetaldehyde are considered with special reference to the comprehensive qualitative estimation of these toxicants and their pathomorphological effects in the target organs. The influence of ethanol-oxidizing enzyme systems in the brain on the development of alcohol tolerance is illustrated. The mechanisms of hormonal regulation via the hypothalamic-pituitary-adrenal axis in the case of exogenous alcoholemia are considered. The dependence of pathological morphological changes in the brain, hypophysis, and adrenal glands on the stage and severity of alcoholic intoxication has been elucidated. Criteria for the morphological and histochemical evaluation of the degree of alcohol-induced lesions and the severity of abstinence syndrome have been developed. The role of alcohol effects in tanatogenesis associated with alcohol-induced diseases is discussed.

Comparison of vasopressin and oxytocin expressions in the hypothalamo-neurohypophysial system of patients with chronic heart failure.

The hypothalamic nonapeptide vasopressin is a known player in the pathogenesis of chronic heart failure. According to the large body of clinical evidence, vasopressin has an impact on salt and water imbalance, hyponatremia, and subsequent renal insufficiency - the most common and destructive co-morbidity of patients afflicted with chronic heart failure. Despite the well-documented elevated levels of vasopressin in the blood of such patients, its expression in the magnocellular hypothalamic nuclei and transport to the posterior pituitary has not yet been investigated. In addition, the literature almost lacks the information on the contribution of another member of nonapeptide family, oxytocin, in the pathogenesis of this disease. Here we present a postmortem analysis of vasopressin and oxytocin-immunoreactive neurons and their terminals in the posterior pituitary of 8 male patients (53.8+/-9.3 years) who had died from CHF and 9 male controls (54.6+/-11.8 years). In line with previous clinical reports, our study on hypothalami of chronic heart failure patients revealed a significant increase in the relative profile density (+29%) of vasopressin-positive neurons in the hypothalamic supraoptic nucleus. Consistently we found a significant increase in the relative optic density of vasopressin-immunoreactivity in the posterior pituitary (+33%) of these patients. In contrast, the similar analysis applied for oxytocin neurons revealed no statistically significant differences to controls. In conclusion, our study provides the morphological evidence for activation of vasopressin (but not oxytocin) expression and vasopressin transport to the posterior pituitary in patients with chronic heart failure.

[Supraoptic nucleus of hypothalamus in subjects with chronic heart failure].

Postmortem paraffin hypothalamic blocks from male subjects with chronic heart failure (CHF) were used for morphometric and immunohistochemical studies of the supraoptical nucleus (SON). Morphometric analysis revealed a significant enlargement of pericarya and nuclei in SON of CHF subjects compared with controls. In addition, eccentrically positioned nucleoli were more frequently found in SON of the subjects with CHF. Moreover, a significant rise in relative entropy of all studied parameters of SON was documented in CHF subjects. The immunohistochemical study revealed a substantial increase of vasopressin immunoreactivity in SON neurons of the CHF subjects in comparison with control ones. Taken together, these results suggest considerable enhancement of synthetic activity of SON neurons in patients with CHF consistent with clinical observations that demonstrate significant elevation of blood vasopressin levels in patients with chronic cardiac insufficiency.

Altered hypothalamic-pituitary-adrenal axis activity in patients with chronic heart failure.

Neuroendocrine factors play an important role in the pathogenesis of chronic heart failure. Despite numerous clinical and experimental studies, the role of the hypothalamic-pituitary-adrenal axis and glucocorticoid hormones is not fully characterised. Here we present a study of plasma cortisol concentration in 74 chronic heart failure patients, divided into four groups based on NYHA functional classes I-IV, and in 17 control subjects. In parallel, we performed morphological analysis of the hypothalamic-pituitary-adrenal axis components from 8 male patients who had died from chronic heart failure, and 9 male controls. In our study we applied immunohistochemical method and quantitative analysis to investigate an expression of hypothalamic neurohormones (corticotropin-releasing hormone, vasopressin) and adrenocorticotropin hormone in the pituitary, as well as performed general histological examination of the adrenal cortex. Measurement of morning cortisol concentration in plasma of chronic heart failure patients revealed neither difference compared to controls nor with the severity of the disease. Despite this, a two-fold increase in the density of corticotropin-releasing hormone-immunoreactive neurons as well as a two-fold increase in the number of corticotropin-releasing hormone neurons co-expressing vasopressin in the hypothalamic paraventricular nucleus were found. In the anterior pituitary the density of adrenocorticotropin hormone-immunoreactive cells was significantly increased. General histological analysis of the adrenal cortex revealed a drastic thinning of the zona fasciculata and dystrophic changes in corticocytes. Structural changes, observed in the adrenal cortex, suggest a relative glucocorticoid deficiency, which may contribute to corticotropin-releasing hormone and adrenocorticotropin hormone upregulation in hypothalamus and pituitary of chronic heart failure patients.

Effects of chronic alcoholic disease on magnocellular and parvocellular hypothalamic neurons in men.

Although numerous data showing severe morphological impairment of magnocellular and parvocellular hypothalamic neurons due to chronic alcoholic consumption have been gathered from animal experiments, only one study (Harding et al., 1996) was performed on POST MORTEM human brain. This study showed a reduction in the number of vasopressin (VP)-immunoreactive neurons in the supraoptic (SON) and paraventricular (PVN) nuclei, but did not provide any data regarding the effect of chronic alcohol intake on human parvocellular neurons. In order to assess whether the changes observed in the animal model also occur in humans and provide a structural basis for the results of clinical tests, we performed immunohistochemical and morphometric analysis of magnocellular (VP and oxytocin, OT) and parvocellular (corticotropin-releasing hormone, CRH) neurons in post-mortem brains of patients afflicted with chronic alcoholic disease. We analyzed 26-male alcoholics and 22 age-matched controls divided into two age groups--"young" (< 40 yr) and "old" (> 40 yr). Hypothalamic sections were stained for OT, VP, and CRH. The analysis revealed: 1) decrease in VP-immunoreactivity in the SON and PVN as well as OT-immunoreactivity in the SON in alcoholic patients; 2) increase in OT-immunoreactivity in the PVN; 3) increase in CRH-immunoreactivity in parvocellular neurons in the PVN. Furthermore, the proportion of cells containing CRH and VP was increased in alcoholics. These findings indicate that chronic alcohol consumption does indeed impair the morphology of magnocellular neurons. The enhancement of CRH-immunoreactivity and increased co-production of CRH and VP in parvocellular neurons may be due to a decline in glucocorticoid production, implied by the hypoplasic impairment of adrenal cortex we observed in alcoholics during the course of this study.

[Diagnostic role of organ-specific isoforms of lactate dehydrogenase in blood plasma].

Changes in plasm isoenzymic spectrum of lactate dehydrogenase (LDG), a glycolytic enzyme, were studied in death of cardiovascular diseases, ethanol poisoning and a mechanical injury. The tests used electrophoresis in 1% solution of agarose. Isoenzymic composition differed between the groups with acute coronary failure, chronic coronary heart disease, alcohol cardiomyopathy and a mechanical injury. Ethanol poisoning caused two types of characteristic shifts in LDG isoenzymic spectrum. Nomograms were designed for practical application in forensic medical expertise of sudden death.

[The activity of ethanol- and aldehyde-oxidizing enzymes of the brain in ethanol intoxication]. / Aktivnost' étanol- i al'degidokisliaiushchikh fermetov golovnogo mozga pri otravlenii étilovym spirtom.

The distributed activity of alcohol dehydrogenase (ADG) and aldehyde dehydrogenase (AldDG) was histochemically studied in neurons of locus coeruleus, nuclei of raphe, substantia nigra and in cerebellar cortex in death or intoxication of ethanol (IE). The enzymatic activity was found to be changing depending on whether there was IE or not. Both higher ADG and lower AldDG were registered in the examined brain sections, which differ by mediatory structure of the neuronic centers. The detected regularities can be used within the forensic medical diagnosis of IE.

[Determination of the severity of endogenous intoxication at the level of average molecular peptides]. / Opredelenie tiazhesti éndogennoi intoksikatsii po urovniu srednemolekuliarnykh peptidov.

A possibility was demonstrated of using the method related with determination of the level of average-weight molecules or average molecules (AM) for the diagnosis of sudden death in forensic medical expertise of cadaver. Blood plasma was investigated according to the method of M.I. Gabriekyan modified by Yu. V. Pervushin, which involves deproteinization of samples with subsequent spectrophotometry at 254 and 280 nm. The AM levels were studied in different pathologies including cardiovascular diseases, alcoholic and narcotic intoxications and pneumonia, caused by a variety of agents, as well as the syndrome of sudden children's death. An analysis of the obtained results denotes a possibility of using the postmortem determination of the AM level concurrently with determination of an endogenous intoxication degree in the forensic medical practice for establishing the cause and mechanisms of nonviolent death.

[Ethanol-oxidizing enzymes of the adrenal glands in acute alcoholic intoxication]. / Etanolokisliaiushchie fermenty nadpochechnikov pri ostroi alkogolnoi intoksikatsii.

Complex morphological-and-histological examinations of the adrenal glands were performed on actual forensic-medical samples in acute alcoholic intoxication (AAI), i.e. 107 death cases of men and women, aged 17 to 60, due to mechanical trauma, lethal ethanol intoxication and somatic complications of chronic alcoholism. A differentiated distribution of ethanol-oxidizing enzymes was defined, i.e. of alcohol-dehydrogenase and acetyl-dehydrogenase in the structural-and-functional zones of the adrenal cortex and medullary substance. A differential activity of enzymes was established that correlated with the level of exogenous alcoholemia. Sets of morphological signs related with examinations of the adrenal glands, which enabled a differential diagnosis of death causes in AAI, were specified.

[Acetaldehyde and some biochemical parameters in alcoholic intoxications]. / Atsetal'degid i nekotorye biokhimicheskie parametry pri alkogol'nykh intoksikatsiiakh.

The need in comprehensive gas chromatography and biochemistry examinations is grounded for cadaver expertise in order to cope with issues related with alcoholic intoxication. Descriptions of 3 examination methods of biological fluids are elucidated, i.e. gas chromatography, electrophoresis and fixing of a degree of endogenous intoxication. The concentration of acetaldehyde in 3 body media (blood, urine and liquor) are analyzed in detail; the isoenzyme spectra of lactate-, alcohol- and aldehyde dehydrogenase as well as the contents of medium molecules in death of alcohol poisonings and due to mechanical trauma are also in the focus of attention.

[Pathomorphological manifestations of various forms of alcohol disease]. / Patomorfologicheskie proiavleniia razlichnykh form alkogol'noi bolezni.

In acute poisoning with ethanol a cardiac variant of tanatogenesis prevails. Signs for ethanol surrogates of DIC syndrome are more characteristic. Alcohol cardiomyopathy and liver cirrhosis are typical as causes of death for chronic forms of alcoholic disease. Histochemical features in the brain are found characteristic for some forms of alcoholic disease.

[Postmortem detection of hemp intoxication]. / Ustanovlenie intoksikatsii preparatami konopli pri sudebno-meditsinskoi ékspertize trupa.

The diagnostic tools applicable to the forensic medical expertise of cadavers in cannabinoid intoxication (CI) were evaluated. Histochemistry with incubation of stable blue "B" (SBB) were used for the detection of cannabinoids in the bronchi and lungs. Hyperemia and capillarostasis in the mucous tunics of the mouth and stomach as well as hyperemia and edema in the lungs and brain were histochemically detected in CI. The histochemical activity of aldehyde dehydrogenase (AlDG) was most essentially decreasing in the adrenal glands and brain of younger persons. The SBB reaction in the bronchial and alveolar epithelia was positive in 85% of cases. A high proof value of the CI diagnostics was pointed out in cadaver expertise.

[The diagnostic value of determining the level of acetaldehyde in blood, urine, and cerebrospinal fluid]. / Diagnosticheskoe znachenie opredeleniia atsetal'degida v krovi, moche i likvore.

The contents of acetaldehyde (AA) in biological fluids obtained from the dead with the confirmed lethality causes, i.e. ischemic heart disease (IHD), alcoholic cardiomyopathy (ACMP) and mechanical traumas (MT), were examined on an actual forensic-medical material (AFMM). 14 death cases of males, aged 18 to 45, were studied. The method of gas-liquid chromatography (GLC), or rather its variation of vaporphase analysis, was used to state the presence and to assess the concentration of acetaldehyde. The results revealed differences between concentrations of acetaldehyde in the examined groups depending on the presence or absence of alcoholemia. Thus, the AA concentrations were found in trace quantities in the MT group free of alcoholic intoxication; while, when it was present in this group, the concentrations went up several-fold. A higher AA content was typical of the ACMP group in all examined subjects both with and without alcoholic intoxication. The final study results are suggestive of that the AA determination in blood, urine and liquor by GLC could be used, within the forensic medical practice, in assessing a severity degree of alcoholic intoxication while establishing the lethal outcome cause due to chronic pathologies and MT.

[Cerebral alcohol dehydrogenase-- a marker of individual ethanol tolerance in ethanol intoxication]. / Alkogol'degidrogenaza mozga--marker individual'noi tolerantnosti k étanolu pri alkogol'noi intoksikatsii

Brain compartments differing by topical chemical location of alcohol dehydrogenases (ADH) were studied on forensic medical material. ADH were found in magnocytes of giant-cell reticular formation, neuronal nuclei in the blue spot, and capillaries, but not in neurons of the ganglionary layer of the cerebral limbic cortex. ADH activity in the studied brain compartments depended on the level of exogenous ethanolemia. Increased ADH activity in the reticular formation magnocytes characterized the individual tolerance in severe ethanol intoxication. Cerebral ADHs are markers of ethanol intoxication.