[The detection of Leiden mutation using technique of enzymatic extension of allele-specific primer with double bioluminescent detection (PED-Biolum)].

The article deals with comparing technique of detection of Leiden mutation on the basis of PEXT-reaction with subsequent bioluminescent microanalysis of products with technique based on RT-PCR. The sampling for testing comprised 83 specimen of genome DNA including 35 specimens with known Leiden heterozygote mutation. The commercial kit "SNP-express-PB" (Litex) was used as a comparison test. It is demonstrated that proposed approach is a simple in its application, effective and relatively inexpensive technique of detection of Leiden one-nucleotide polymorphism in gene V of blood coagulation factor. The technique "PED-Biolum" has no differences in comparison with commercial technique RT-PCR concerning ability to detect mutant allele and matches it in parameters of economic effectiveness.

[The phenotype IFNy+cd69- of T-lymphocytes of umbilical blood associated with epidemiologically verified risk factors of atopy formation].

The article deals with the determination of possible role of imbalance of lymphocytes TH1/TH2 at birth in pathogenesis of atopy development in children. The subpopulation compound of umbilical blood lymphocytes was examined on sampling of 153 newborns. The following prospective monitoring was applied during first year of life. It is established that the low content ofnonactivated mononuclears in umbilical blood producing gamma-interferon (IFNy+/CD69-) is associated with atopic family anamnesis (p = 0.006), residing of pregnant woman in town (in comparison with rural area, p = 0.02) and male gender of newborn in case of repeated labor (p = 0.008). This situation matches the epidemiologic verified risk factors of atopy formation in children. The contact of pregnant woman with domestic pets results in increasing of content of activated lymphocytes producing gamma-interferon (IFNy+/CD69-), p = 0.03. The decrease of content of nonactivated cells producing gamma-interferon (IFNy+/CD69-) is a strong and independent predictor of formation of atopic dermatitis in children of infancy age. The relative risk of formation of atopic dermatitis consisted 3.4 (1.19-9.69).

[Value of enzyme-linked immunosorbent assay for detection of p24 antigen of human immunodeficiency virus in confirmation of HIV-infection].

AIM: To evaluate diagnostic value of p24 antigen detection for algorithm of confirmatory diagnostics of HIV-infection. MATERIALS AND METHODS: Concurrently with Western blot assay (WB, "New Lav Blot1", Bio-Rad), tests for detection of p24 antigen of HIV (Genetic Systems HIV-1 Ag EIA", "VectoHIV-1 p24-antigen confirming test", and "DS-EIA-HIV-AG-SCREEN") were used for confirmation of first-positive result of immuno-enzyme assay. RESULTS: p24 HIV antigen was detected in serum samples in 8.4% of patients with equivocal result of WB and in 4.2% of patients with negative and positive results of WB. Presence of p24 was correlated with high viral load, and, in patients with confirmed diagnosis, with low CD4 cells count (<500 cells/ml). p24 was detected in more than 30% of persons with confirmed seroconversion after primary testing. CONCLUSION: In groups of persons with negative and equivocal results of WB assay, detection of HIV p24 antigen points to the presence of infection and could be the reason for the final diagnosis. Detection of p24 in patients with positive result of WB assay allows to consider them as probable candidates for highly active antiretroviral therapy.

[The clinical value of immunological markers of EBV infection in children with infectious mononucleosis].

A hundred and sixteen patients aged 1 to 14 years who had infectious mononucleosis (IM) caused by the Epstein-Barr virus (EBV) were followed up in its acute phase and 3, 6, and 12 months after the disease experienced by them. The clinical significance of the markers of EBV infection was assessed in children with IM. The high diagnostic capacity of detection of EBV DNA in the lymphocytes was established in patients with IM. It was shown that it is necessary to use the serological markers of EBV infection to monitor the course of this disease in children.

[Evaluation of the metabolic state and functional activity of immunocompetent cells in children with relapsing herpetic viral infection]. / Otsenka metabolicheskogo sostoianiia i funktsional'noi aktivnosti immunokompetentnykh kletok u detei s retsidiviruiushchei gerpesvirusnoi infektsiei.

Specific features of the immune response were studied (during the clinical remission period) in 40 children (aged 3 to 6) with a relapsing herpes viral infection (RHVI). The following was established: an increased relative and absolute quantity of lymphocytes, a reduced percentage content of CD+ and an increased absolute count of CD+ in the peripheral blood. Changes in the immunity humoral chain manifested themselves by an increased content of class G immunoglobulins and of circulating immune complexes (CIC) in the blood serum. A reduced quantity of phagocyting blood neutrophils was registered with the absorbing ability being preserved. Changes in the metabolic status of immune-competent cells were established; they were described by a high activity of the plastic exchange reaction, a tensed energetic potential and a reduced glutathione-recovery reaction modulating the proliferative activity of lymphocytes. A reduced reserve potential of oxygen-dependent biocenosis of neutrophyl granulocytes was detected in the peripheral blood. Obviously, the structural and metabolic changes in the blood lymphocytes as well as the abnormalities in the functional activity of neutrophyl granulocytes, observed during the clinical remission period in the RHVI period, are a cause predetermining the relapses of the disease and can be regarded as a criterion for applying the immune rehabilitation.

[Diagnostic significance of chemiluminescent response of neutrophil granulocyte in children with lymphadenopathy]. / Diagnosticheskoe znachenie pokazatelei otveta neitrofil'nykh granulotsitov u detei s limfadenopatiiami.

Chemiluminescent response of the blood neutrophilic granulocytes was evaluated during the diagnostic period in 22 patients with Hodgkin's disease, 35 with nonspecific lymphadenopathies, and 24 patients with tuberculous infection aged 7-14 years. Disorders in spontaneous and induced chemiluminescence of neutrophils were detected in all patients, and were the most pronounced and stable in patients with Hodgkin's disease. Evaluation of neutrophilic chemiluminescent response in these patient groups leads to a more accurate diagnosis of lymphadenopathies of different origin.

[Clinical significance of NADPH-dependent dehydrogenases in lymphocytes from patients with bronchial asthma]. / Klinicheskaia znachimost' izmenenii aktivnosti NADF-zavisimykh degidrogenaz v limfotsitakh u bol'nykh bronkhial'noi astmoi.

The activities of lymphocyte NADP-dependent dehydrogenases were evaluated in 84 children with atopic asthma during relapses and clinical remission. The pattern and severity of changes in the studied enzymes depended on children's age and duration and severity of disease. Changes in the older age group and during relapse were the deepest and most stubborn. During remission the activities of the studied oxidoreductases did not reach the parameters observed in healthy children. Changes in the activities of NADP-dependent dehydrogenases in blood lymphocytes objectively reflect the course of allergic inflammation in patients with atopic asthma and can be clinically used for evaluation of the pathological process at a metabolic level.

[Specific antigenic preparations as inducers of blood leukocyte respiratory burst in chemiluminescent analysis]. / Ispol'zovanie spetsificheskikh antigennykh preparatov v kachestve indukatorov dykhatel'nogo vzryva leikotsitov krovi pri khemiliuminestsentnom analize.

Functional activity of blood leukocytes of normal subjects and patients with herpesvirus infection was studied by the chemiluminescent method in vitro with specific antigenic preparations (tick-borne encephalitis vaccine, ADPT vaccine, and herpetic vaccine). Functional response of blood leukocytes to vaccine preparations is specific and depends on the antigen and metabolic status of cells of recipients. The range of chemiluminescent response of blood leukocytes is wide: no changes in tick-borne encephalitis vaccine, weak response to herpetic vaccine, and pronounced changes in functional activity in response to ADPT vaccine. Testing with different doses of specific inducers helps additionally evaluate the metabolic reserve of blood leukocytes. A pronounced response of leukocytes to specific antigenic preparations in vitro (e.g. to ADPT vaccine) prompts development of methods for predicting the reaction of nonspecific reactivity system to vaccination.

[The antihypoxic and radioprotective properties of selective adenosine receptor agonists]. / Protivogipoksicheskie i radioprotektornye svoistva selektivnykh agonistov adenozinovykh retseptorov.

The selective A1- and A2-receptor agonists L-PIA (ED50 = 0.55-0.78 mmole/kg) and NEGA (D50 = 0.05-0.1 mmole/kg) possess high antihypoxic and radioprotective effects. A2-subtype receptors may be involved in the antihypoxic and radioprotective effects of adenosine analogs.

[Biochemico-pharmacological mechanisms and the relation between the anticalorigenic, hypothermic and antihypoxic effects of adenosine]. / Issledovanie biokhimiko-farmakologicheskikh mekhanizmov i vzaimosviazi antikalorigennogo, gipotermicheskogo i antigipoksicheskogo éffektov adenozina.

Administration of adenosine into mice induced a decrease in oxygen consumption (anticalorigenic effect), body temperature (hypothermal effect) and protected against acute hypoxia (antihypoxic action); all these effects are closely correlated. Isopropyl noradrenaline and theophylline prevented but propranolol and dipiridamol increased these effects of adenosine. Being administered into mice adenosine decreased both cAMP content (in skeletal muscles and kidney) and respiration in kidney homogenates. These adenosine effects appear to be realized via the following chain: A1-receptors----decrease in cellular cAMP content----inhibition of tissue oxygen consumption----development of anticalorigenic reactions----hypothermal effects----antihypoxic actions.

[Effect of emotional stress on the activity of enzymes of glutathione metabolism]. / Vliianie émotsional'nogo stressa na aktivnost' fermentov metabolizma glutationa.

Emotional stress during 20 min induced the stimulation of the glutathione peroxidase activity in heart, liver and kidney tissues and the activation of glutathione-S-transferase in heart and liver tissues. gamma-Glutamyl transferase activity was inhibited in liver and kidney tissues. The alterations were gradually decreased in heart greater than or equal to liver greater than kidney. Both catecholamines (in vivo) and cAMP (in vitro) produced the same effects. Close correlation between the stress and cAMP actions on the glutathione metabolizing enzymes was found. No changes of glutathione reductase activity was observed in all the tissues studied.

[Protective effect of adrenergic alpha 2-receptor agonists in hypoxic hypoxia]. / Zashchitnoe deistvie agonistov alpha 2-adrenoretseptorov pri gipoksicheskoi gipoksii.

The antihypoxic effect of alpha 2-adrenoceptor agonists was studied by two different approaches: reproduction of the effect by a number of alpha 2-agonists and its blockade with selective antagonists. The data obtained suggest that alpha 2-adrenoceptor agonists increase the survival and the lifespan of mice in all the models of acute hypoxic hypoxia under study. A close correlation between antihypoxic action of alpha 2-adrenoceptor agonists and their anticalorigenic effect was established (r = +0.87; P less than 0.01).