[Antimycotic therapy impact on oral mucosa Candida species composition in HIV-infected patients]. / Vliianie antimikoticheskoi terapii na vidovoi i shtammovyi sostav gribov roda Candida polosti rta bol'nykh VICh-infektsiei.

RESEARCH OBJECTIVES: the analysis of a specific and strains drift of Candida in HIV/AIDS patients with oropharyngeal candidiasis and the analysis of Candida sensitivity dynamics to reference antimycotic drugs. The study comprised 49 HIV-infected patients aged 20-69 years. The study revealed candidiasis treatment provides specific and strains drift of Candida. Eradication of fluconazole sensitive C. albicans leads to growth of more resistant strains (C. glabratae, krusei, tropicalis) thus lowering antimycotic therapy efficacy. The efficacy improvement requires selective approach to candidiasis treatment with azol agents.

[Prevalence of DNA viruses in maxillofacial area of HIV-infected]. / Porazhennost' DNK-virusami cheliustno-litsevoi oblasti bol'nykh s VICh-infektsiei.

DNA viruses have high oncogenic risk viruses; they cause emergence of Kaposi sarcoma, Lymphoma, Squamous cell carcinoma. HIV immunodeficiency promotes increase in frequency of such tumors. Etiotropic therapy of HIV patients considerably reduces prevalence of DNA viruses and a viral malignization.

[Specific features of lymphoid infiltration in liver biopsy in patients with chronic viral hepatitis B and C].

AIM: To study characteristics of leukocytic infiltration of the liver in patients with chronic viral hepatitis B and C (CVHB and CVHC) with consideration of hepatitis activity and fibrosis severity. MATERIAL AND METHODS: The examination of 37 patients with CVHB (n = 13) and CVHC (n = 24) included liver puncture biopsy by Mengini, subsequent histological and morphological investigation of liver biopsy with immunohisto- and immunocytochemistry. RESULTS: In CVHB and CVHC patients leukocytic infiltrates (LI) of the liver present primarily with T-lymphocytes (CD3+), NKT cells (CD3+CD16+CD56+), NK cells (CD16+CD56+), T-regulatory lymphocytes (CD4+CD25+), cytotoxic T-lymphocytes (CD8+). Cytotoxic lymphocytes (CD8+) and NK cells (CD16+CD56+) detected in hepatic LI of patients with chronic viral hepatitis are similar in composition with cells in hepatic tissue infiltrates in autoimmune hepatitis. We are the first to detect the complex of T-regulatory cells (CD4+CD25+) in hepatic parenchyma of these patients. This complex suppresses cellular immune response in virus elimination and damaged tissues and supports development of persistent viral infection with autoimmune component. CONCLUSION: The complex of T-regulatory cells (CD4+CD25+) isolated in hepatic LI evidences for existence of a morphofunctional base for autoimmune manifestations in the presence of persistent viral infection.

[Approaches to improvement of laboratory diagnosis of infectious diseases].

Modern routine and novel techniques for diagnostics of infectious diseases and identification of intoxication syndrome markers are reviewed. Also presented are results of original studies of mixed intestinal infections and methods of their rapid differential diagnostics.

[Lymphocyte subpopulation composition in hepatic tissue and autoimmune manifestations in viral hepatitis].

This work had the aim to study the role of disbalance of inflammatory procytokines, lymphocyte subpopulation composition in peripheral blood, and cell populations of hepatic tissue in pathogenesis of viral hepatitis. A total of 159 patients with chronic hepatitis B and C were examined. They proved to exhibit signs of autoimmune processes (ASMA, ANA, AMA, anti-LKM-1, RF, CG) and cytokine (IL-1b, TNFa) disbalance. The number of lymphocytes expressing HLADR+ was shown to increase and CD+ lymphocyte to decrease in proportion to the severity of periportal necrosis, portal inflammation, intralobular degeneration and fibrosis compared with healthy donors. Hepatic biopsies contained T-regulatory cell complex (CD4+ CD25+). These data give evidence of the morphofunctional basis for autoimmune manifestations in patients with persistent viral hepatitis infection.

[Characteristics of the cytokine profile and peripheral blood lymphocyte subpopulations in patients with chronic viral hepatitis B and C in comparison to severity of morphological changes of the liver].

AIM: To study cytokine concentration in blood serum in spontaneous and NDV- and PGA stimulated production by peripheral blood mononuclear cells (PBMC) and contents of peripheral blood lymphocyte subpopulations when compared with severity of morphological changes of the liver in patients with chronic viral hepatitis B and C (CHB and CHC). MATERIAL AND METHODS: A total of 37 patients entered the trial (13 with CHB and 24 with CHC). Concentrations of IFN-alpha, IFN-gamma, TNF-alpha, IL-1beta, IL-4, IL-6 in blood serum in spontaneous and NDV and PGA stimulated PBMC production, content of CD3, CD4, CD8, CD16, CD25, HLADR, CD20, CD38 subpopulations of peripheral blood lymphocytes were examined. Also, puncture biopsy of the liver by Mengini was made with subsequent estimation of the histological activity index by R. Knodell adapted to hepatitis B and C by K. Ishak. RESULTS: Patients with CHB and CHC in minimal activity of the process and insignificant morphological changes in the liver demonstrated a significant fall of NDV induced IFN-alpha production. Patients with highly active hepatitis and maximal morphological changes in the liver had significant lowering of NDV induced production of IFN-alpha, IFN-gamma, TNF-alpha, IL-6 and PGA induced production of TNF-alpha. CONCLUSION: CHB and CHC patients should be examined not only with estimation of serum cytokines concentration but also NDVand PGA stimulated production of PBMC should be taken into consideration with calculation of immunoregulatory index for prognosis of the disease and choice of adequate therapy.

[T-cell responsiveness to specific group A streptococcus antigens in patients with recurrent erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in 51 patients with recurrent erysipelas versus 63 patients with primary erysipelas. To reveal LMA, the authors employed in vitro a screening cell migration test as an indicator of the cooperation of T and B lymphocytes and macrophages, by stimulating with polysaccharide A, surface proteins, L-antigen, hyaluronidase, streptolysine-O, and a complete set of Grasse S. pyogenes. In patients with recurrent erysipelas, undulating LMA changes were found in the course of the disease in response to the stimulation with partial specific S. pyogenes antigens. There were differences in the time course of LMA changes, when stimulated with specific surface streptococcus antigens and with components of streptococcus with toxic activity. Significant LMA differences were found in relation to the pattern of a local process: active LMA changes from acceleration to suppression in the erythematous-hemorrhagic form and hyperergic reactions of LMA acceleration in the bullous-hemorrhagic one.

[Detection of clostridia toxin markers in different types of the course of acute intestinal infections].

The purpose of the investigation was to study the detection rates of markers and the level of C. diffcile A and B toxins and C. perfringens type A enterotoxin in patients with acute intestinal infections (AII). Two hundred and seventy-three patients with AII of varying etiology were followed up. According to the clinical syndrome, the patients were divided into 3 groups: (1) patients with the gastroenteritic (GE) type; (2) those with the gastroenterocolitic (GEC) type; (3) those with enterocolitic (EC) type. The circulation of markers of C. difficile A and B toxins and C. perfringens type A enterotoxin was studied, by employing the immunological test systems in the coagglutination test using the plates. The higher levels of antigens of all toxins were identified in the acute period of the disease in the GE and EC types than in the GEC type. There was a short increase in the levels of antigens of the test toxins in the GEC type and a gradual decrease in the GE and EC types. By discharge from hospital, the markers of toxins (more commonly of C. diffcile A) were preserved in 16.4% of the patients mainly in the GEC type.

[Pathogenetic aspects of intoxication syndrome in the clinical picture of infectious diseases].

Pathogenetic mechanisms of intoxication syndrome in patients with infectious diseases are considered in the context of current general pathologic concepts. All major components of pathogenesis are discussed in terms of the authors' original observations and literature data. Bacterial endo- and exotoxines are believed to be the main inductors of intoxication. Results of original studies suggest an important contribution of Shiga toxins to specific clinical manifestations of infectious diseases. A novel interpretation of the role of intoxication syndrome in the development of infectious diseases is proposed.

[Differential sensitivity of peripheral blood granulocytes, monocytes, and lymphocytes to endotoxin-induced apoptosis in patients who have sustained Salmonella infection].

The differential sensitivity of peripheral blood granulocytes, monocytes, and lymphocytes to endotoxin-induced apoptosis was studied in convalescents with Salmonella infection. The number of early (Tunel+PI-) and late (Tunel+PI+) apoptotic cells and the rate of apoptosis (the relative content of DNA apoptotic fragments in the solitary cell) were determined on a FACSCalibur cytofluorometer, by applying a package of the CellQuest programs (Becton Dickinson). A higher threshold of sensitivity of granulocytes, monocytes, and lymphocytes to the apoptotic effect of low, suboptimal doses (100 ng), as well as suppression of early stages of apoptosis by the optimal doses (1000 ng) of S. enteritidis lypopolysaccharides (LPS) were observed in patients who had sustained salmonellosis at a stage of reconvalescence. The hierarchy of LPS-induced apoptosis (granulocyte--monocyte--lymphocyte), observed in healthy donors, was also retained in the group of patients receiving the suboptimal doses of LPS, which seems to be associated with the repeated action of toxin on sensitized cell populations.

[T-cell responsiveness to specific group A streptococcus antigens in patients with primary erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in 63 patients with primary erysipelas. To reveal LMA, a screening cell migration test (SCMT) was used as an indicator of the cooperation of T- and B-lymphocytes and macrophages in the stimulation with polysaccharide A, surface proteins, L-antigen, hyaluronidase, streptolysin O, a complete S. pyogenes antigen complex after Grasse. The prognostic value of MAL parameters was established at week 1 after the onset of erysipelas. A rapid transition of LMA from the phase of acceleration to that of inhibition was shown to characterize the formation of an adequate response, to correspond to the good course of the disease, and to be followed by the low likelihood of recurrences. The probability of a subsequent recurrence is much higher than that when LMA tends to transit from suppression to acceleration and when LMA parameters are constant in the phase of suppression or acceleration. No transition of LMA to the phase of suppression in early convalescence suggests that the formation of an immune response to streptococcus is delayed.

[Lipopolysaccharide-induced apoptosis of peripheral granulocytes in patients with prior salmonella infection].

The in vitro induction of granulocytes in the presence of high and low concentrations of lipopolysaccharides (LPS) allowed the authors to study a dose-dependent apoptotic response in the population of intact cells and in that of primed cells in the period of convalescence in patients with Salmonella infection. An experimental system was used to study a respiratory burst and granulocytic apoptosis in the presence of hypothalamic neuropeptides. The clinical manifestations of the diseases were studied in 32 inpatients. It was shown that there was an increase in the sensitivity of granulocytes to the apoptotic action of low, suboptimal doses (100 ng) of S. enteritidis LPS in patients with salmonellosis at convalescence. This correlates with the severity of a clinical course of the experienced disease and with the suppressed sensitivity of apoptosis to the optimal doses (1000 ng) LPS.

[Time course changes in cytokines content and gas composition in blood of patients with salmonellosis and acute shigellosis].

AIM: To study changes in serum concentrations of interleukines (IL) and tumor necrosis factor alpha (TNFa) in the course of acute moderate and severe shigellosis and salmonellosis; to elicit their correlation with disorders of gas and electrolyte blood composition and acid-base balance (ABB). MATERIAL AND METHODS: A total of 39 patients with salmonellosis and 32 patients with acute shigellosis admitted to infectious hospital N 2 entered the study. The following parameters were assessed: serum concentrations of IL-1, IL-IO and TNFa; ABB, gas and electrolyte blood composition; leucocytic intoxication index (LII). The tests were made on the disease day 2-3 and 6-7 (in severe salmonellosis on day 10-11). RESULTS: In moderate salmonellesis and acute shigellosis the level of proinflammatory cytokines (IL-1 and TNFa) diminished while in severe acute intestinal infection their concentration was high reflecting imbalance of immune response. Content of IL-10 depends on etiology and severity of the course of infectious process - the highest IL-10 concentrations were found in patients with severe salmonellosis on the disease day 10-11. A direct correlation was confirmed between IL-1, TNFa and LII in the disease onset in all patients with acute intestinal infections. CONCLUSION: An important role of IL-1, IL-10 and TNFa in pathogenesis of bacterial intestinal infections is confirmed. A correlation exists between blood gas composition and concentration of cytokines.

[Clinical features and current laboratory diagnosis of leptospirosis of grippotyphosa serogroup].

AIM: To characterize persistence of the causative agent in the blood of patients with leptospirosis, to evaluate significance of the results of polymerase chain reaction (PCR) with isolation of ribosomal RNA (rRNA) and microagglutination reaction (MAR) in diagnosis of leptospirosis at different stages of the disease. MATERIAL AND METHODS: The study included 94 patients with leptospirosis: 51 (54.2%) males, 43 (45.8%) females, 74 (78. 7%) urban and 20 (21.3%) rural citizens in acute leptospirosis, early and late convalescence. Leptospirosis was diagnosed on the basis of clinico-epidemiological, laboratory-device data. All the patients were divided into 2 groups: with leptospiral jaundice (n = 33, 35.1%) and without leptospiral jaundice (n = 61, 64.9%). The disease of moderate severity was in 64 (68%) patients. Verification of the diagnosis was based on determination of specific antibodies titer in MAR using standard set of diagnostic cultures. PCR determined rRNA of leptospira in the blood on the disease day 5, 10 and 15, 1, 2, 3, 6 and 12 months after the disease onset. The results were processed with methods of parametric and non-parametric statistics. CONCLUSION: PCR demonstrated diagnostic value in early leptospirosis (sensitivity of the method in the disease week 1 was 86%). Diagnostic significance of MAR reached 81.8% later - in 3 weeks. Detection of leptospiral rRNA by PCR in the autopsy material allows one to recommend this method for additional verification of the diagnosis including as an alternative to bacterioscopic test.

[A clinical and pathothysiological study of protein C in patients with generalized meningococcosis]. / Kliniko-patofiziologicheskoe issledovanie proteina C u bol'nykh s generalizovannoi formoi meningokokkovoi infektsii.

The clinical-and-pathogenetic significance of protein C was dynamically investigated within the infection processes in 23 patients (mean age 44 years) with meningococcemia. A reliably lower concentration of protein C in blood plasma (mean 37.8%) was registered versus the normal parameters (N = 70-130%) at exacerbation irrespective of a disease outcome. Leyden mutation was detected in 30.5% of patients. The presence of the above genetic defect denoted a predisposition to a severe disseminated intravascular coagulation (DIC) syndrome in patients of the studied group. A lower concentration of protein C was detected in blood plasma of patients with meningococcemia; it correlated with severity of the infection process and with the development of organic malfunction. The study of the concentration of protein C enabled an evaluation of the functional condition of anticoagulation mechanisms in the DIC development and can be regarded as an important extra criterion for the evaluation of thrombohemorrhagic syndrome in patients with meningococcemia.

[Humoral and cellular adaptation mechanisms in development of intoxication syndrome in patients with acute viral hepatitis]. / Gumoral'nye i kletochnye adaptatsionnye mekhanizmy pri razvitii intoksikatsionnogo sindroma u bol'nykh ostrymi virusnymi gepatitami.

AIM: To study endotoxin-binding function of blood in patients with acute viral hepatitis (AVH) in the course of the disease. MATERIAL AND METHODS: A total of 335 patients with AVH were examined. The titers of IgG antiendotoxin antibodies to glycolipid of Re-chemotype were estimated in enzyme immunoassay (Diaendotox-2 kit), endotoxin-binding activity (EBA) of high density lipoproteins (HDLP) in EIA with the kit Liprobind (Microecos). The number of polymorphonuclear leukocytes (PL) binding lipopolysaccharides were counted by M. Yu. Yakovlev et al. method. For detection of PL binding endotoxin of gram-negative bacteria, kits Endim (Microecos) were used, beta2-microglobulin was estimated by a radioimmunoassay using kits made in Belarus Institute of Bioorganic Chemistry. Registration of chemoluminescence was conducted on chemiluminometer LBK 1251 (Finland) by M. Wilson (1985) method. RESULTS: In moderate AVH at the height of intoxication a low Re-antibodies level was recorded irrespective of the disease etiology. To convalescence this level went up. PL reserves in AVH patients were low except patients with AVH C who had unaffected neutrophil reverves in relation to LPS binding. At the height of AVH, HDLP EBA was very high in all the groups with normalization to convalescence. In VHC high HDLP EBA persisted as long as the disease course. A significant rise of beta2-microglobulin was seen in all the groups of AVH patients, its severity correlating with the degree of the disease severity. CONCLUSION: One of the leading factors in development of indoxication in AVH patients is a lipopolysaccharide complex of gram-negative intestinal bacteria actively coming into blood flow due to hepatocyte cytolysis. In response to LPS action there appeared a stereotypic adaptation complex of reactions in the form of activation of cellular and humoral immune response.

[Cell-cooperation integrated test for the detection of immune reactivity to Shiga-toxin in patients with acute intestine infection]. / Integrativnyi test kletochnoi kooperatsii dlia vyiavleniia immunnoi reaktivnosti k shiga-toksinu u bol'nykh ostrymi kishechnymi infektsiiami.

Described are the results of approbation of the cell-migration screening test reflecting the outcome of cooperation between T- and B-cells and macrophages. It was shown, in adults and children with intestine infection, to be a highly-effective tool for the detection of suppressed immune response during early disease stages at stimulation in vitro by Shiga toxin at nano- and picogram concentration; it can also be used for the evaluation of the shaping specific anti-Shiga-toxic immune response. The parameters of the migration activity of peripheral-blood leucocytes at exacerbation and convalescence were demonstrated to correlate with the age of sick children and with the severity of intestine infection as well as with the level of a Shiga-like toxin detected in coprofilters and circulating immune complexes.

[Autoimmune manifestations in acute viral hepatitides]. / Autoimmunnye proiavleniia pri ostrykh virusnykh gepatitakh.

AIM: To study autoimmune reactions in patients with acute viral hepatitis A, B, and C in the dynamics of an infectious process. MATERIALS AND METHODS: 134 patients with acute viral hepatitides of various etiology were examined. Among them there were 36 patients with acute viral hepatitis A, 40 with B, and 58 with viral hepatitis C. Antinuclear antibodies (ANA), antismooth muscle antibodies (SMA), and antimitochondrial antibodies (AMA) were determined by immunofluoresence assay using the test systems ATS COMVI-1, IMMCO diagnostics (USA). Serum rheumatoid factor was detected by the semiquantitative latex-agglutination test on a slide manufactured by the "Human" firm (Germany). Cryoglobulins were identified by the qualitative visual method. RESULTS: SMA, ANA were detected in the diagnostically significant titers (1:40-1:160) in patients with acute viral hepatitis A, B, and C. A distinction of autoimmune manifestations was ascertained in viral hepatitis C. It has been demonstrated that in 71% of the patients cryoglobulins appear in the acute period of the disease and, in their presence, rheumatoid factor was found in 17.2% and 6.9% exhibited anti-LKM-1 and AMA. CONCLUSION: The presented data on the nature and time course of changes in autoimmune reactions in patients with acute viral hepatitis A, B, and C during an infectious process suggest that autoimmune disorders are characteristic of viral hepatic lesions just at the early stages of the disease irrespective of its etiology and play a prominent role in the pathogenesis of these nosological entities.

[Characteristics of cellular immunity in children with acute viral hepatitis A]. / Kharakteristika pokazatelei kletochnogo immuniteta u detei, bol'nykh ostrym virusnym gepatitom A.

AIM: To study the populational composition of lymphocytes and the specific features of production of cytokines in children with acute viral hepatitis A (AVHA). MATERIALS AND METHODS: 128 patients aged 11 to 14 years who had AVHA, moderate AVHA being in 83.5% were examined. In 87.2% of the children, the disease was cyclic. The etiology of the disease was verified by simultaneously detecting anti-HAV IgM (enzyme immunoassay) and by the presence of HAV RNA (polymerase chain reaction) in the blood. Peripheral lymphocytes (CD) were phenotypes in the indirect immunofluorescence test using monoclonal antibodies; cytokines were determined by the enzyme immunoassay; serum beta 2-microglobulin was done by radioimmunoassay. RESULTS: The children with AVHA were found to have elevated levels of tumor necrosis beta-factor, interleukin-1 beta (IL-1 beta), and IL-4 in the icteric period, as well as a decrease in their levels at convalescence. IL-6 was detected in individual patients only in the first 3 days of the icteric period. The peak of AVHA was characterized by relative lymphocytosis, by decreases in the counts of T helper/inductor cells and natural killer cells, by increases in the count of CD25 cells, convalescence, by preserved lymphocytosis, by the increased levels of T lymphocytes that carry the markers CD3, CD4, CD8, CD16, CD25, and CD95, and by higher toxicity of beta 2-microglobulin.