[Development of the quantitative immunochromatography tests for somatic deasease markers detection].

The purpose of work was creation of quantative immunochromatographic tests (ICT) for measuring concentrations of the marker substance associated with somatic diseases: immunoglobulin E (IgE), C-reactive protein (CRP) and fibrin D-dimer in blood serum (plasma), which is carried out with the help of videodigital analyzers of domestic development "Reflecom" and "Zondaj". ICT were designed in sandwich-format, using colloid gold and monoclonal antibodies. It is shown, that calibration curves received with the help of ICTand devices of videodigital registration, are well approximated by exponential dependence Y = Aexp(-x/B)+y0, where Y--device readings, x--analytes concentration, A, B, y0--constants. Samples of blood serum (plasma), under investigation received in conditions of a hospital from patients. Correlation with electrochemiluminescent immunoassay, enzyme linked immunoassay, latex-agglutinations assay and immunochromatographic method was observed. Sensitivity of the developed tests was 7,5 IU/ML for IgE, 5,7 ng/ml for CRP, 500 ng/ml for fibrin D-dimer. The developed analytical complex- videodigital analyzer "Zondaj" and ICT for quantitative measure of concentration IgE, fibrin D-dimer and CRP- can be successfully applied in laboratory practice and clinical laboratory researches alongside with actual immunochemistry methods.

[Indication of extremely dangerous infectious pathogens using immunochromatography and digital video analysis].

The use of immunochromatographic indicatory elements based on antibody conjugates and colloidal gold was suggested to detect cells and the antigens of extremely dangerous infectious pathogens. The specificity and specific activity (sensitivity) of the mentioned elements were studied on vaccinal strains of plague, anthrax, and tularemia pathogens. The researchers studied a possibility to increase the sensitivity of immunochromatographic analysis using computed scanning and Reflecom, a specialized digital video recorder of immunochromatogramms. The study demonstrated that the use of electronic devices to record immunochromatogramms increased the sensitivity to microbial cells and antigens 1.5 to 2 times vs. visual registration and simplified documentation of the results.

Suppression of alcohol consumption in C57Bl/6 mice after intranasal administration of antiserotonin antibodies.

Experiments on C57Bl/6 mice under conditions of free choice between alcohol and water for 10 months showed that intranasal administration with antiserotonin antibodies in doses of 30 and 15 microg/kg for 2 week decreased alcohol consumption over 1 month after treatment.

Immunization of rats with conjugates of dopamine and serotonin with bovine serum albumin prevents the development of experimental MPTP-induced depressive syndrome (electrophysiological parameters).

Electrophysiological experiments on Wistar rats demonstrated that prior immunization of animals with conjugates of dopamine and serotonin with bovine serum albumin, as well as with bovine serum albumin alone, played a partial protective role in relation to the subsequent development in these animals of experimental MPTP-induced depressive syndrome: immunized animals showed no signs of the depressive state such as decreases in the latency of onset of REM sleep and the development of epileptiform activity in the caudate-putamen complex, though the increase in the proportion of REM sleep in the overall structure of sleep persisted. Changes in the spectral characteristics of brain electrical activity and sleep structure during the development of experimental MPTP-induced syndrome in animals immunized with conjugates of dopamine and serotonin with bovine serum albumin and with bovine serum albumin alone were antigen-specific and reflected functional shifts in the activity of those neurotransmitter systems targeted by immunization, as well as others sensitive to changes in the body's immunological status.

Antibodies to dopamine as neuromodulators of behavioral responses of mice of different genotypes.

The possibility of long-term changes in the innate characteristics of the behavior of mice of different genotypes (C57Bl/6 and BALB/c) after active immunization with dopamine-BSA conjugates was studied. These experiments revealed significant differences in the effects of anti-dopamine antibody on behavioral responses in an open field and neurotransmitter contents of brain structures in mice of different lines. Immunization with the dopamine-protein conjugate led to increases in the functional activity of the corresponding brain receptors, to different extents in mice of different lines.

[Immunization of rats with bovine serum albumin conjugate with dopamine or 5-hydroxytryptamine prevents the development of experimental MPTP-induced depressive syndrome. An electrophysiological study]. / Immunizatsiia krys kon"iugatami dofamina i serotonina s bych'im syvorotochnym al'buminov predotvrashchaet razvitie éksperimental'nogo MFTP-indutsirovannogo depressivnogo sindroma (po élektrofiziologicheskim pokazateliam).

In electrophysiological experiments in Wistar rats it was shown that preventive immunization of animals with bovine serum albumin conjugated with dopamine or 5-hydroxytryptamine or with bovine serum albumin alone partly protects against the development of experimental MPTP-induced depressive syndrome. Signs of depressive-like state such as decrease in REM-sleep latency and the development of epileptiform activity in caudate putamen were not observed in immunized animals, whereas REM-sleep proportion in the total sleep structure was increased. Changes in the spectral characteristics of brain electric activity and sleep structure during development of experimental MPTP-induced syndrome in immunized rats are antigen-specific and reflect the functional shifts in neurotransmitter systems, both in those which are the target of the immune action and those sensitive to alterations in the immune state of a body.

Immunomodulation of inherent behavior in C57Bl/6 and BALB/c mice with antibodies to glutamate.

Immunization of BALB/c mice with glutamate-BSA conjugate reduced anxiety and improved passive avoidance retention, while in C57Bl/6 mice immunization disturbed passive avoidance retention, but had no effect on anxiety. Interstrain differences in the shuttle box behavior were found between control animals.

Antibodies to glutamate as neuromodulators of behavioral reactions in mice with various genotypes.

We studied the effects of active immunization with glutamate-protein conjugates on behavioral reactions of C57Bl/6 and BALB/c mice in the open-field test and on animal anxiety in the tail suspension test. Antiglutamate antibodies produced different behavioral changes in mice of these strains.

[Dopamine antibodies as neuromodulators of behavioral reactions in mice of different genotypes]. / Antitela k dofaminu kak neiromoduliatory povedencheskikh reaktsii myshei raznykh genotipov.

A possibility of the long-term modification of inborn behavioral features of different mice genotypes (C57B1/6 and BALB/c) by active immunization with dopamine-bovine serum albumin conjugate was investigated. Significant interstrain differences were found in the effects of dopamine antibodies on the open-field behavior and the content of neurotransmitters in the brain cortex and striatum. It was shown that the active immunization of mice to dopamine produces an increase in the functional activity of brain dopamine receptors. The extent, to which this increase is pronounced, is genotype-dependent.

Neuromodulatory effects of serotonin antibodies on behavioral responses, neurotransmitter content in the central nervous system, and ligand-binding activity of central serotonin receptors in C57BL/6 and BALB/c mice.

Effects of active immunization with the serotonin-protein conjugate on open-field behavior, learning and retention of the conditioned passive avoidance response, neurotransmitter contents in brain structures, and functional activity of central serotonin receptors were studied on C57Bl/6 and BALB/c mice. Serotonin antibodies produced opposite effects on the studied parameters in genetically different mouse strains.

[Alteration in the brain electrical activity, diurnal sleep structure, and the rat behavior after immunization with bovine serum albumin conjugated with dopamine]. / Izmeneniia élektricheskoi aktivnosti mozga, struktury dnevnogo sna i povedeniia krys posle immunizatsii zhivotnykh kon''iugatom dofamina s bych'im syvorotochnym al'buminom.

Male Wistar rats (380-430 g) were immunized with bovine serum albumin conjugated with dopamine (2 mg/kg, 0.25 ml) mixed with Freund's adjuvant complete (0.25 ml) or with bovine serum albumin mixed with Freund's adjuvant complete in the same doses. One week after the immunization with bovine serum albumin conjugated with dopamine, irregular spike activity and high-amplitude spindles associated with the state of awake immobility were recorded in the rat neocortex and caudate putamen, the relative power of the electrical activity in the caudate putamen was decreased in the alpha band, while the relative power of the beta 1 in the cortical EEG was increased. In the structure of 4-hour diurnal sleep, a decrease in the mean duration of sleep episodes and a reduction in the REM sleep content were observed. The parameters of the electrical activity and diurnal sleep structure returned to normal during the following 4 weeks. The open-field behavior 2 weeks after the second immunization (without Freund's adjuvant complete) did not differ from that of the control rats immunized only with bovine serum albumin. Titres of antibodies to dopamine after the second antigen injection were 1:32-1:64 in the electrophysiological series and 1:128-1:256 in behavioral experiments.

[Alterations in diurnal sleep structure, electrical activity, and neurochemical parameters of the rat brain induced by systemic injection of complete Freund adjuvant and immunization by bovine serum albumin with complete Freund adjuvant]. / Izmeneniia struktury dnevnogo sna, élektricheskoi aktivnosti i neirokhimicheskikh pokazatelei mozga krys pri sistemnom vvedenii zhivotnym polnogo ad''iuvanta Freinda i pri immunizatsii syvorotochnym al'buminom s polnym ad''iuvantom Freinda.

Investigations of the effects of animal immunization with immunogenesis stimulator Freund's adjuvant complete (alone or in combination with bovine serum albumin often used in control experiments) on brain electrical activity, sleep, and neurochemical parameters were carried out in male Wistar rats. It was shown that both injection of Freund's adjuvant complete alone (0.25 ml) and immunization with bovine serum albumin (2 mg/kg in 0.25 ml of saline) mixed with Freund's adjuvant complete (0.25 ml) led to an increase in the slow-wave and REM sleep. After injection of Freund's adjuvant alone, development of sleepiness was gradual and reached its maximum within 3-5 weeks, while after the combined treatment the alterations in the sleep structure became pronounced already 1 week after the first antigen injection and persisted at least for 5 weeks. Neurochemical analysis revealed no significant changes in the noradrenaline, dopamine, and serotonin content in striatum and frontal neocortex after the injection of Freund's adjuvant. After the combined treatment, the serotonin content in these structures decreased. After the Freund's adjuvant injection, the dynamics of changes in power spectra of the brain electrical activity of different brain structure in the state of quiet wakefulness was complicated. Increase in the slow-wave activity in the delta 1 range (1-2 Hz) in caudate putamen, basomedial nucleus of amygdala, and sensorimotor cortex was observed in the animals immunized with bovine serum albumin mixed with Freund's adjuvant complete 1 week after the antigen injection and later on during the whole observation period. This was probably associated with an adaptive increase in the functional activity of serotoninergic system.

[Modulation of behavior of mice of different genotypes by serotonin antibodies]. / Moduliatsiia antitelami k serotoninu povedencheskikh reaktsii myshei raznykh genotipov.

The effect of active immunization with conjugated serotonin-protein on the "open-field" behaviour and passive avoidance conditioning was studied in genetically different mice strains (C57BL/6 and BALB/c). The obtained immunophysiological effects of serotonin antibodies depended on genetically determined characteristics of animal behaviour. Serotonin antibodies altered rearing and crossings of the "open-field" center and retention of passive avoidance learning in C57BL/6 mice. The active immunization with conjugated serotonin-protein of BALB/c mice resulted in modulation of the "open-field" latency and both training and testing of passive avoidance behaviour.

Effect of antiserotonin antibodies on bioelectrical activity of sensorimotor cortex.

The effect of antiserotonin antibodies on basal electrocorticogram was studied in electrophysiological experiments on rats. Intracortical injection of 10 microg antiserotonin antibodies into the sensorimotor cortex induced epileptiform activity in this area. It is assumed that antiserotonin antibodies modulate activity of cortical neurons due to both binding serotonin molecules and interaction with serotonin receptors.

Formation of morphine tolerance in offspring of morphine-tolerant animals: neurochemical and neuroimmune correlates.

We carried out a complex physiological, neurochemical, and neuroimmunologic study of the formation of tolerance to analgetic effect of morphine and analyzed enkephalinase A activity in different brain structures and serotonin antibodies in the serum. More early development of morphine tolerance and a sharp increase in serum antibody titer was found in the offspring of morphine-tolerant rats. This points to an imbalance in the neurotransmitter system and can serve as a diagnostic marker of endogenous opioid system pathology.

Anti-dopamine antibodies: effects on behavior in an "open field," pain sensitivity, CNS monoamine content, and functional activity of immunocytes in C57Bl/6 mice.

Single i.p. doses of anti-dopamine antibodies were given to C57Bl/6 mice. This resulted in inhibition of motor activity in a large proportion of the animals in the "open field" test, which lasted five days. Hyperalgesia, detected 1.5 h and 1 day after doses of antibody, was replaced by analgesia on day 5. There was a sharp reduction in the levels of dopamine and its metabolites in the cerebral cortex at 1 and 5 days; the serotonin level was increased 1 day after doses of antibody, and was significantly decreased at 5 days. There was no effect on cells of the immune system. The possible mechanisms of the neurotropic action of these antibodies are discussed.

[Effect of immunizing rats with a serotonin-bovine serum albumin conjugate on the structure of diurnal sleep and electrical activity of the brain]. / Vliianie immunizatsii krys kon''iugatom serotonin-bychii syvorotochnyi al'bumin na strukturu dnevnogo sna i élektricheskuiu aktivnost' mozga.

Rats were immunized with bovine serum albumin conjugated with 5-hydroxytriptamine (5-HT). Two injections with a 2-week interval were carried out. At first, rats were injected subcutaneously (2 mg/kg, 0.25 ml) with Freund's adjuvant complete. The second injections (10 mg/kg intraperitoneally) was made 2 weeks later without the adjuvant. One week after the first antigen injection, there were a decrease in the relative power of the electrical activity of the caudate putamen, neocortex, and amygdala in the delta 2 and theta 2 bands and its increase in the alpha and beta-ranges. The number and duration (total and mean) of REM sleep episodes as well as REM sleep contribution in the total sleep duration decreased. These findings testify to a reduced functional activity of the central serotonergic system. Four-five weeks after the first injection, a tendency to a recovery of the bio-electrical spectral pattern and diurnal sleep structure was revealed. Five weeks after the first antigen injection, the titre of antibodies to 5-HT was 1:32-1:64.

[The effect of active immunization with a conjugate of dopamine and bovine serum albumin on the development of an experimental MPTP-induced depressive syndrome and on the monoamine metabolism in the brain of rats]. / Vliianie aktivnoi immunizatsii kon''iugatom dofamina s bych'im syvorotochnym al'buminom na razvitie éksperimental'nogo MFTP-indutsirovannogo depressivnogo sindroma i obmen monoaminov v golovnom mozge u krys.

Active immunization with dopamine conjugated with bovine serum albumin (DA-BSA) or BSA with complete Freund's adjuvant (CFA) partly suppressed the development of the MPTP-induced depressive syndrome in rats preventing the appearance of "behavioral despair" symptoms: increase in immobility time and higher index of depression in forced-swim test. In DA-BSA-immunized rats the content of DOPA, DA, HVA, NA, and 5-HN in caudate putamen and that of NA in the frontal cortex was increased, while in BSA-immunized rats the content of 5-HT in both brain areas and that of DOPAC in the frontal cortex was decreased both in rats with reduced depressive syndrome and in saline control as compared with intact animals a day after the last drug injection. In DA-BSA-immunized rats with reduced depressive syndrome the increase in DA and 5-HT content in caudate putamen was less expressed and DOPAC content was lower than in saline control. In BSA-immunized depressive rats DA content in the frontal cortex was also reduced as compared to control.

[Effect of dopamine antibodies on neuropathic pain syndrome in rats]. / Vliianie antitel k dopaminu na razvitie neiropaticheskogo bolevogo sindroma u krys.

Dopaminergic brain system plays an important role in regulation of pain sensitivity. However, the data on participation of antidopamine antibodies in the development of neurogenic pain are absent. This work was aimed at the study of the role of antidopamine antibodies in the development of pain syndrome induced by the injury of nn. ischiadic and saphenous in rats. It was shown that after the nerve injury, the behavioral reaction such as autotomy (self-injury) appeared as a feature of the experimental neuropathic pain syndrome. It was originally established that the development of neuropathic pain syndrome induced by the injury of peripheral nerves was accompanied by induction of dopamine autoantibodies. It was also shown that immunization of the animals with conjugated dopamine-protein autigen resulted in aninerease of autidopamine antibody level and an amplification of the experimental neuropathic pain syndrome, i.e., decrease in the latency of the first autotomy, increase in expression of autotomies, and increase in the number of animals with late autotomies.

[Dopamine antibodies: effect on the "open field" behavior, pain sensitivity, monoamine levels in the central nervous system, and functional activity of immunocytes in C57Bl/6 mice]. / Antitela k dofaminu: deistvie na povedenie v "otkrytom pole", bolevuiu chuvstvitel'nost', soderzhanie monoaminov v TsNS i funktsional'nuiu aktivnost' immunotsitov myshei linii C57Bl/6.

Dopamine antibodies (AB) were singularly injected to C57B1/6 mice intraperitoneally. The locomotor activity in the open field was suppressed for 5 days in the majority of the animals. Hyperalgesia revealed 1.5 h and 1 day after the AB injection changed for analgesia on the 5th day. A sharp reduction of the brain level of dopamine and its metabolite was revealed 1 and 5 days after the AB injection, the serotonin content was increased within 1 day and decreased within 5 days after the injection. No action of the AB on cells of the immune system was observed. The possible mechanisms of the AB neurotropic action are discussed.