[Effect of reboxetine on activity of carboxypeptidase E in the nerve tissue of rats].

Depression is one of the most common mental disorders, but its etiology is not completely understood. It is assumed that peptidergic system components are involved in the formation of this pathology. Neuropeptides play an important role in the regulation of mental and emotional states. Сarboxypeptidase E is a key enzyme of peptide processing; it regulates neuropeptide levels in the various structures of the nervous system. We have studied effects of a single dose of reboxetine on the activity of carboxypeptidase E in various brain regions and the adrenal glands of rats. The reboxetine injection decreased carboxypeptidase E activity in the pituitary gland (12 h after injection), in the pituitary gland, the quadrigeminal bodies, the medulla oblongata, the hypothalamus, the hippocampus and the amygdala (24 h after injection), in the pituitary gland and striatum (72 h after injection). The enzyme activity in adrenal glands remained basically unchanged. Apparently, the decrease of carboxypeptidase E activity may influence the level of regulatory peptides involved in the pathogenesis of depression.

[Effect of selank on the main carboxypeptidases in the rat nervous tissue].

Effect of single selank administration on activity of carboxypeptidase H and of phenylmethylsulfonylfluoride-inhibited carboxypeptidase - enzymes detaching arginine and lysine from C-terminus of molecules-precursors of biologically active peptides was studied. The preparation has been shown to cause long, preserved for 24 h changes of activities of these carboxypeptidases. It is suggested that the change in activity of the studied enzymes can be one of mechanisms of regulation of level of neuropeptides at action of selank.

Effect of cholinergic drugs on the activity of basic carboxypeptidases in rat nervous tissue.

Effects of a single administration of cholinergic drugs (arecoline, atropine, nicotine, mecamylamine) on the activity of carboxypeptidase H and of phenylmethylsulfonyl fluoride-inhibited carboxypeptidase, which are involved in metabolism of neuropeptides, were studied in brain parts and the adrenal glands of rats. The enzyme activities were determined fluorimetrically using specific inhibitors and substrates. In the majority of cases the enzyme activities decreased, and this decrease was retained for at least 72 h. Changes in the activities of the studied enzymes depended on the type of cholinergic action, the nervous system part, and the time after the injection. The changes in activities of the studied carboxypeptidases are supposed to be a possible mechanism responsible for changes in the levels of neuropeptides under the influence of high doses of the drugs.

[The blood acid-base values of various qualifying groups of sportsmen in physiological condition and in exercise].

The acid-base parameters and buffer systems of blood of different qualifying groups of sportsmen in physiological condition and at physical work were studied. It is shown that initial differences of the acid-basic condition and buffer capacity of integral blood of sportsmen of high qualifying group lead to changes of the pH dynamics at raising concentration of lactate. Dynamics of the blood lactate concentration is analyzed at stepped-raising exercise in sportsmen of different qualification, the possible molecular mechanism of sharp increase in concentration of lactate in the blood is discussed. It is shown that the point of a break on a curve of change of the pH precedes a point of break on a curve of increase in concentration of lactate at physical work. The received results can be used for definition of the moment of approach of an anaerobic threshold with pH-measurement of the blood.

[Proteolytic activity of placenta with EPH-gestosis].

Essential edema-proteinuria-hypertension (EPH) gestosis still represents an important obstetrical problem. We have investigated the activity of carboxypeptidase H (CPH), phenylmethylsulfonyl fluoride inhibited carboxypeptidase (PMSF-CP), carboxypeptidase M (CPM) and angiotensin-converting enzyme (ACE), the main carboxypeptidases in human placenta under normal conditions and mild EPH-gestosis. Gestosis was accompanied by the decrease in activity of the enzymes involved into metabolism of regulatory peptides (ACE, CPH, PMSF-CP, CPM) compared with their activity in placenta under physiological pregnancy. Correlation analysis revealed positive correlation between placental CPH and CPM (r = 0.2735*) in EPH-gestosis. These findings suggest involvement of placental proteases into formation of compensatory-adaptive reactions in the fetoplacental complex at EPH-gestosis; the data obtained may be also employed for the development of methods of prophylaxis and corrections of metabolic impairments in pathology of pregnancy.

[Effect of arecholine and atropine administration on the carboxypeptidase H and phenylmethylsulfonyl fluoride-inhibited carboxypeptidase activities in the rat nervous tissue].

The effect of a single dose administration of arecholine and atropine on the activities of carboxypeptidase H and phenylmethylsulfonyl fluoride-inhibited carboxypeptidase involved into the final stage of formation of biologically active neuropeptides from precursors has been studied. Changes in the enzyme activities were evident during at least 72 h after administration of these drugs. These results suggest that one of possible mechanisms of reduction of neuropeptide levels by arecholine and atropine consists in suppression of activity of enzymes taking part in there metabolism--carboxypeptidase H and phenylmethylsulfonyl fluoride-inhibited carboxypeptidase.

[Activity of peptidyl-dipeptidase A and carboxypeptidase N in the serum of patients with Alzheimer disease].

The activity of peptidyl-dipeptidase A and carboxypeptidase N taking part in peptide metabolism in the serum of patients with Alzheimer disease were studied. The role of these enzymes in the metabolism of neuropeptides and beta-amyloid at the Alzheimer disease was discussed.

[The effect of prenatal stress on the carboxypeptidase H activity in the hypothalamo-pituitary-adrenal-gonadal system of the rat].

Effects of dams stress on the carboxypeptidase H: the neuropeptide exchange enzyme in the hypothalamo-pituitary-adrenall-gonadal system, was studied in the litter of different age (0, 14, 28, 45, and 120 day after birth) was studied. The sex differences in dynamic of enzyme activity in intact animals are substantiated. The effect of prenatal stress on carboxypeptidase H activity dependes on age and sex of animals. Prenatal stress is altering during the age dynamics of enzyme activity. This in the pituitary gland and hypothalamus of prenatal stressed female rats was a control for the male rats. In the adrenal and gonadal gland of prenatal stressed males, this was a control for the female rats female. The role of carboxypeptidase H in pubescence and mechanisms of effect of prenatal stress on sex system functional are discussed.

[The effect of ethanol consumption by dams on the offspring locomotion in the open field test and carboxypeptidase activities in the rat brain and adrenal medulla].

Consumption of dams ethanol increased the posterity locomotion activity in open field test. The increase in female rats was higher then in male ones. Differences in the carboxypeptidase H and PMSF-inhibited carboxypeptidase activities between the brain regions and adrenal medulla of prenatally exposed to ethanol and intact rats were found. The changing of enzyme activities in female rats was higher then in male ones. It is possible that dams ethanol consumption induced profound changes in locomotion mediated, at least partially, by changes in the rate of proteolytic processing of neuropeptide precursors.

[Atropine effect on activity of angiotensin-converting enzyme and carboxypeptidase n in the serum of rats].

The effect of a single atropine administration on the activities of angiotensin-converting enzyme and carboxypeptidase N taking part in peptides metabolism was studied. Changes in the enzymes activities after administration were evident during at least 72 h. The role of these enzymes in the metabolism of neuropeptides under the effect of atropine was discussed.

[Effect of a single emotional-pain stress on the basic carboxypeptidases activity in the rat brain regions and adrenals]. / Vliianie odnokratnogo émotsional'no-bolevogo stressa na aktivnost' osnovntkh karboksipeptidaz v otdelakh mozga i nadpochechnikakh krys.

Effect of a single emotional-algesic stress on the carboxypeptidase H and PMSF-inhibited carboxypeptidase activities (phenylmethylsulphonilfluoride-inhibited carboxypeptidase) taking part in the final stage of formation of biologically active neuropeptides from precursors, was studied. Activity of the enzymes depended on duration of the stress and time after the stress in pituitary and adrenal glands. Differences in changes of carboxypeptidase H and PMSF-inhibited carboxypeptidase activities was found, especially in adrenals. The role of enzymes in emotional-algesic stress development and in metabolism of regulatory peptides by the stress was discussed.

[Activity of the basic carboxypeptidases in female rat tissues at different stages of estrus cycle]. / Aktivnost' osnovnykh karboksipeptidaz v tkaniakh samok krys na raznykh stadiiakh éstral'nogo tsikla.

It is revealed, that the activity of neuropeptide metabolism enzymes (carboxypeptidase H, phenylmethylsulfonilfluorid-inhibited carboxypeptidase) in the female rat tissues depends upon the stage of estrus cycle. The carboxypeptidase H activity in the pituitary gland is the highest in proestrus; it is almost 3 times higher in comparison with diestrus; it is a little bit higher in striatum on the stage of estrus, than in diestrus and proestrus, in adrenals on the stage of proestrus and estrus it is a little bit lower, than in diestrus; in the ovaries on the stage of proestrus it is much higher, than in estrus and diestrus. The activity of PMSF-inhibited carboxypeptidase in ovaries on the stage of proestrus and diestrus is 1.7-1.8 times higher, than at the stage of diestrus. The activity of carboxypeptidase M in adrenal tissue at the stage of proestrus is 35-40% of that at the stage of diestrus and estrus. The activity of carboxypeptidase M in the ovaries at the stage of diestrus is 45-50% of that at the stage of diestrus and estrus. The role of the investigated enzymes in cyclic changes of a level of biologically active peptides and in regulation of estrus cycle is discussed.

Isolation, partial purification, characterization, and tissue distribution of phenylmethylsulfonyl fluoride-inhibited feline carboxypeptidase.

Phenylmethylsulfonyl fluoride (PMSF)-inhibited carboxypeptidase from cat liver was purified 148-fold by chromatography on CM- and DEAE-cellulose with 27.3% yield. Molecular weight of the enzyme is 100-110 kD as determined by gel filtration on Sephadex G-150. The enzyme has maximum activity at pH 5.50-5.75; its activity is completely inhibited by PMSF or p-chloromercuribenzoate and partially inhibited by iodoacetamide. EDTA, 2-mercaptoethanol, N-ethylmaleimide, Co(2+) and Ca2+, basic carboxypeptidase inhibitor guanidinoethylmercaptosuccinic acid, and angiotensin-converting enzyme inhibitor captopril do not influence its activity. The enzyme cleaves arginine from enkephalin-Leu5-Arg6 and dansyl-Phe-Leu-Arg to form enkephalin-Leu5 and dansyl-Phe-Leu, respectively, and very slowly cleaves leucine from carbobenzoxy-Gly-Leu. Further cleavage of either enkephalin-Leu5 or dansyl-Phe-Leu was not detected. The highest activity of this enzyme was found in adrenal glands and testicles; this activity was 30% lower in hypophysis, and still lower in liver and kidney. The PMSF-inhibited carboxypeptidase activity in brain was about 6-16 times lower than that in adrenal gland. In brain regions, the highest activity was detected in gray matter of cerebral hemispheres and cerebellum, and slightly lower activity was found in thalamus/hypothalamus, striatum, and hippocampus. The lowest activity was found in quadrigeminal bodies, medulla oblongata, and white matter of cerebral hemispheres. The enzyme exists mainly in soluble form; the activity of membrane-associated enzyme is 7-25% of soluble enzyme activity depending on tissue type. We consider here a possible involvement of PMSF-inhibited carboxypeptidase in the metabolism of biologically active peptides.

[Effect of testosterone and progesterone on carboxypeptidase H activity during stress in mice]. / Vliianie testosterona i progesterona na aktivnost' karboksipeptidazy H pri stresse u myshei.

It was found that testosterone propionate (3 mg/kg of body weight) and progesterone (1 mg/kg of body weight) partially prevent an augmentation of the carboxypeptidase H activity in the mouse pituitary gland under stress caused by a single intraperitoneal administration of olive oil. In testicles, testosterone prevented augmentation of the enzyme activity within 0.5 hours but increased it through in 4 and 24 hours after treatment. Progesterone in testicles augmented the enzyme activity within 0.5 hours, but not in 4 and 24 hours. Sex steroids were not affected by stressinduced carboxypeptidase H activity in hypothalamus and adrenal medulla. It was concluded that carboxypeptidase H did not take part in integration of hypothalamus-pituitary-adrenal and hypothalamus-pituitary-gonadal axis under the stress.

[Effect of chloroform and ether narcosis on the activity of basic carboxypeptidases in the hypothalamic-pituitary-adrenal axis in rats]. / Vliianie khloroformnogo i éfirnogo narkoza na aktivnost' osnovnykh karboksipeptidaz v gipotalamo-gipofizarno-nadpochechnikovoi sisteme krys.

It is discovered that chloroform narcosis does not influence on carboxypeptidase H and phenylmethylsulfonyl fluoride-inhibited carboxypeptidase activity in the rats hypothalamic-pituitary-adrenal axis. Ether narcosis provokes an increase of PMSF-inhibited carboxypeptidase activity in the pituitary body approximately in 8 times and carboxypeptidase H activity in hypothalamus by 29 percents in comparison with the intact animals. It is supposed that at research neuropeptides and their metabolism enzymes and especially the answer to a stress chloroform narcosis would be the better anaesthesia method than ether narcosis.

[Effect of chronic ethanol consumption on basic carboxypeptidase activity in regions of the rat brain]. / Vliianie khronicheskogo potrebleniia étanola na aktivnost' osnovnykh karboksipeptidaz v otdelakh mozga krys.

It is discovered that chronic consumption of ethanol induced decrease of carboxypeptidase H activity in striatum by 27%; increase of carboxypeptidase M activity in hippocampus by 67% and decrease in cerebral hemispheres by 34%; phenylmethylsulfonyl fluoride-inhibited carboxypeptidase activity increase in hypothalamus by 141%, in striatum by 60% and in optic and lamina quadrigemina by 34%. The role of basic carboxypeptidases in mechanisms of ethanol influence on the peptidergic systems are discussed.

[Determination of angiotensin-converting enzyme activity using dansyl-Phe-Ala-Arg as the substrate]. / Opredelenie aktivnosti angiotenzinprevrashchaiushchego fermenta s ispol'zovaniem dansil-Phe-Ala-Arg v kachestve substrata.

Modificated method for the determination of the angiotensin converting enzyme (EC 3.4.15.1) activity in neural tissue is proposed. The methods is based on fluorimetrically determination of released dansyl-Phe from dansyl-Phe-Ala-Arg at pH 7.6. Km is 50 +/- 10 MM. The high specificity of the method is provide by using captopril, the high specific angiotensin converting enzyme inhibitor. The sensitivity of the method is 0.001 nmol/min per mg protein that it is lesser in 15-fold than the lowest enzyme activity in brain region. The distribution of the angiotensin converting enzyme activity that using the method in brain regions and peripherial tissues of rat is presented.

[Effect of testosterone on carboxypeptidase H activity in the murine hypothalamus and hypophysis]. / Vliianie testosterona na aktivnost' karboksipeptidazy H v gipotalamuse i gipofize myshei.

Effects of single intraperitoneal administration of testosterone propionate (3 or 30 mg/kg body weight) on activity of carboxypeptidase H in pituitary body and hypothalamus of female white mouse were studied. It was found that testosterone propionate treatment (3 and 30 mg/kg body weight) increased the carboxypeptidase H activity in pituitary through 0.5 hour and it decreased one in 24 hours after treatment. The carboxypeptidase H activity in hypothalamus was lower as compared with control animals in 24 hours after testosterone propionate treatment in the dose 3 mg/kg body weight. However, the carboxypeptidase H activity in hypothalamus was lower in 0.5 and 24 hours and it was higher in 4 h after testosterone propionate treatment in the dose 30 mg/kg body weight as compared with the control. These data suggest that testosterone affects the carboxypeptidase H activity by changing the level of enzyme gene expression.

[Effect of testosterone and progesterone on carboxypeptidase H activity in the hypothalamo-hypophyseal-gonadal axis and the adrenal gland in mice]. / Vliianie testosterona i progesterona na aktivnost' karboksipeptidazy H v gipotalamo-gipofizarno-gonadal'noi osi i nadpochechnikakh myshei.

Testosterone propionate decreased activity of carboxypeptidase H in the male mongrel mice pituitary gland, activity of the CP H in testicles, and increased these parameters in 4 hours after treatment. Progesterone decreased the CP H activity in pituitary gland and increased it in testicles. Progesterone increased the CP H activity in female pituitary gland and decreased it in adrenal gland and hypothalamus. The role of the CP H in the effects of the sex steroids on the function of peptidergic systems is discussed.