[Identification of Shiga toxin in patients with acute intestinal infections in the presence of mono- and mixed-O-antigens of pathogens]. / Vyiavlenie Shiga-toksina u bol'nykh ostrymi kishechnymi infektsiiami v prisutstvii mono- i mikst-O-antigenov vozbuditelei.

AIM: To investigate the time course of changes in the detection rates and levels of Shiga toxin antigen (STA) in their stool and middle-molecule circulating immune complexes (CICs) containing IgG (IgG CIC) in patients with acute intestinal infections (AIIs) in the presence of the body's circulation of mono- and mixed-LPS/O-antigens of intestinal pathogens. SUBJECTS AND METHODS: A total of 147 patients aged 15 to 55 years who had been hospitalized with AIIs were examined. The diagnosis was bacteriologically verified in 19% of the patients; in the others, it was confirmed by the detection of LPS/O-antigens of Shigella, Salmonella, Yersinia, and Campylobacter in their stool by means of the reaction of coagglutination (RCA) on glass slides. Plates for RCA displayed STA in the fecal and IgG CIC samples. RESULTS: Mono- and mixed infections were detected in 32 and 68%, respectively. The RCA plates exhibited STA in 25.2% of the fecal samples and in 90.5% of the IgG CIC ones from patients with AIIs and did not in those from donors. In monoinfection, the detection rates and levels of STA in the feces became lower in the course of the disease and remained unchanged in IgG CIC and the levels of STA also decreased in the feces, but increased in IgG CIC in mixed infection. CONCLUSION: In 25.2% of the patients with early AIIs, their stools show free STA; its detection rate and levels are significantly higher in mixed infections than those in monoinfection. The level of STA in serum IgG CIC was significantly higher in mixed infection, suggesting an active immune response to the pathogen. Given that the Shiga toxin-producing strains are present in patients with AIIs, caution should be exercised in the choice of an antibacterial drug to prevent horizontal gene transfer and to enhance toxin production and the body's intoxication. One of the advantages of RCA is the possibility of rapidly changing the spectrum of test systems, depending on the region of their application and the epidemiological situation.

[Markers of intestinal infection agents in patients with acute viral hepatitis with cholestatic syndrome].

AIM: To study the pathogenetic role of agents of the most common intestinal infections in the development of cholestatic syndrome in patients with acute viral hepatitis. MATERIAL AND METHODS: We investigated 97 patients with acute viral hepatitis (AVH), 112 blood donors and 165 patients with acute bacterial intestinal infections (ABII). O-antigens of the major causative agents of infections in feces and serum (CIC) and the presence of antishigatoxic CIC in the serum were found. RESULTS: It is first shown a high incidence of O-antigens of Salmonella, Yersinia, Helicobacter, Shigella and Campylobacter in 81.4% of patients with AVH, more frequent and prolonged their detection in patients with the syndrome of cholestasis, higher total O-antigenic load, the frequent identification of mixed-antigens, high levels of LII in comparison with patients without AVH cholestasis and with acute intestinal infections patients. One-third of patients with AVH revealed antishigatoxic CIC in blood serum, which is lower than in patients with ABII. CONCLUSION: The findings suggest the importance of intestinal infections agents in the development and maintenance of inflammation in the liver, the prognostic value of identifying markers in patients with AVH, especially with the syndrome of cholestasis.

[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.

[Postinfection irritable bowel syndrome].

AIM: To characterize pathogenesis, clinicolaboratory criteria and treatment of postinfection irritable bowel syndrome (PICS). MATERIAL AND METHODS: The examination including histological study of the small and large intestine mucosa, polymerase chain reaction (PCR), coagglutination reaction using shigella, salmonella, yersinia, campilobacter jejuni diagnosticums, indirect hemagglutination reaction for identification of antibodies to these agents in the blood serum was conducted in 750 patients with PICS. Fecal seeding on selective media was made as well as the respiratory test for bacterial growth in the small intestine. Immune status was studied with laser cytometry, chemiluminescence, immunodiffusion, immunofluorescence, flow laser cytofluorometry. Personality profile was assessed by MMPI. RESULTS: PICS was diagnosed in 599 (79.9%) of 750 patients. Most of them had diarrhea, abnormal fecal microflora, antigens of acute intestinal infection agents in circulating immune complexes of the serum and coprofiltrates. Immune system was characterized by low phagocytic activity, attenuation of cell and humoral immunity. Etiotropic and pathogenetic treatment including intestinal antiseptics, probiotics and immunomodulators produced persistent remission during a year in 79.3% PICS patients. CONCLUSION: PICS is described which differs from ICS by registration of markers of acute intestinal infections in biological media, bacterial overgrowth in the small intestine and dysbiosis in the large intestine, immunodeficiency. A positive response was observed to treatment with intestinal antiseptic and enterosorbent drugs, probiotics and immunomodulators.

[Prognostic value of leukocytic migratory activity indices in patients with erysipelas].

Peripheral blood leukocytic migratory activity (LMA) was studied in patients with primary or recurrent erysipelas. A screening cell migration test (SCMT) was used in vitro and it established the prognostic value of MAL parameters at week 1 after the onset of erysipelas. It has been shown that a rapid transition of LMA from the phase of acceleration to that of inhibition characterizes the formation of an adequate immune response, corresponds to the good course of the disease, and has a low likelihood of recurrences. The probability of a 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.

[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.

[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.

[Occurrence of Helicobacter pylori specific antigens in diseases of gastrointestinal tract]. / Chastota vstrechaemosti spetsificheskikh antigenov Helicobacter pylori pri zabolevaniiakh zheludochnokishechnogo trakta.

The results of the immunophoretic analysis of H. pylori antigenic structure are presented. Along with H. pylori O-antigen, specific surface antigen of acidic nature, relatively thermolabile with characteristics similar to those of K-antigens, was detected. A diagnosticum based on the coagglutination test for the rapid detection of H. pylori specific antigens in the patients' biological fluids (coprofiltrate, saliva) by the noninvasive method was developed. The circulation mode of H. pylori in human body and its correlation with the clinical symptoms of the disease was shown. H. pylori antigens were very frequently detected (in 65 - 83% of cases) in patients with acute bacterial enteric infections: shigellosis, salmonellosis, yersiniosis, campylobacteriosis. The newly developed method seems to be promising for the rapid diagnosis of H. pylori associated infections, for the prolonged monitoring and evaluation of the effectiveness of antibacterial therapy.

[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.

[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.

[The pathogenetic and clinico-diagnostic significance of changes in leukocyte migration activity in patients with shigellosis and salmonellosis]. / Patogeneticheskoe i kliniko-diagnosticheskoe znachenie izmenenii migratsionnoi aktivnosti leikotsitov u bol'nykh shigellezami i sal'monellezami.

The peripheral blood leukocyte migration activity (LMA) was studied in 134 acute dysentery patients and 129 salmonellosis patients by using the in vitro screening assay of cell migration (SACM) from microcultures, stimulated with LPS antigens from a number of enteric pathogens. At the acute period of the disease (days 1-5) in the greater part of the patients (80-90%) the specific acceleration of LMA, later followed by its inhibition, was registered in the presence of the pronounced intoxication and diarrhea syndromes, as well as the intensive circulation of specific antigens of the infective agents in the body (established by the method of the coagglutination test). The established phase fluctuations of LMA correlated with the severity and character of the infectious process. SACM was found to be highly sensitive and specific, convenient, well reproducible and having good prospects for the early diagnosis of enteric infections, including those of mixed etiology.

[The screening detection of the antigens of pathogenic enterobacteria in the biological fluids of patients with chronic intestinal diseases]. / Skriningovoe vyiavlenie antigenov nekotorykh patogennykh énterobakterii v biologicheskikh zhidkostiakh u bol'nykh s khronicheskimi zabolevaniiami kishechnika.

Screening detection of the antigens of some pathogenic enterobacteria in the coagglutination test with the use of Shigella (S. sonnei, S. flexneri 2a, 1b, 6), Salmonella (groups A, B, C, D, E) and Yersinia (Y. pseudotuberculosis, serovars I and III, Y. enterocolitica 03 and 09) diagnostica in patients with chronic enteric diseases, hospitalized in a noninfectious ward, has demonstrated a high incidence of the above antigens. S. flexneri antigens have shown the highest detection rate. The results thus obtained indicate that these infective agents probably take part in the development of exacerbations of chronic enteric diseases and, therefore, a comprehensive examination of this group of patients is necessary with a view to the detection of bacterial enteric infection.

[The detection of the Shiga toxin antigen in connection with other virulence factors--the O and K antigens of enterobacteria]. / Vyiavlenie antigena shigatoksina v sviazi s drugimi faktorami virulentnosi--O- i K-antigenami énterobakterii.

The results of the detection of Shiga toxin antigen in culture filtrates of 73 enterobacterial strains by the methods of countercurrent electrophoresis and coagglutination, as well as their correlation with the presence of other factors of virulence, O- and K-antigens, have been analyzed. Sharply pronounced inverse correlation between the capacity of enterobacteria to synthesize Shiga (Shiga-like) toxins and their capacity to synthesize acidic polysaccharide surface-specific K- (Vi-) antigens has been established.

[The rapid diagnosis of salmonellosis by a determination of the antigen in the biosubstrates of patients using the coagglutination reaction]. / Uskorennaia diagnostika sal'monelleza opredeleniem antigena v biosubstratakh bol'nykh s pomoshch'iu reaktsii koaggliutinatsii.

The diagnostic potential of the coagglutination test was checked with the aim of improving the laboratory diagnosis of Salmonella infections by the detection of Salmonella specific antigen in different biological materials (feces, urine, saliva and immune complexes in blood sera). The study of all specimens resulted in the confirmation of the diagnosis in 78% of patients, often during the first days of the disease. The proportion of nonspecific reactions, as shown in the control groups of healthy donors and patients with dysentery and other acute enteric infections, did not exceed 5%.

[Latent persistence of specific antigens of the causative agent in the composition of circulating immune complexes in typhoid fever]. / Skrytoe persistirovanie spetsificheskikh antigenov vozbuditelia v sostave tsirkuliruiushchikh immunnykh kompleksov pri briushnom tife.

This study revealed the presence of O-antigen of group D salmonellae and Vi-antigen in circulating immune complexes in patients with typhoid fever, bacteriologically confirmed (56 +/- 5.6% and 65 +/- 5.4% of cases, respectively) and not confirmed (15.5 +/- 5% and 39 +/- 7% of cases, respectively), in patients with diarrhea of nontyphoid etiology in the presence of negative results of the coagglutination test and in healthy persons. The level and dynamics of circulating immune complexes were established with respect to the antigens contained in these complexes. Altogether O- and Vi-antigens were detected in circulating immune complexes, respectively, in 92%, 96% and 94% of cases on weeks 1, 2 and 3 from the beginning of the disease, and in all cases on weeks 4 and 5, the antigens being determined together and separately. Thus, the latent persistence of S. typhi antigens as part of circulating immune complexes in the blood serum was established. The determination of such persistence is of great pathogenetic and diagnostic importance, which also applies to the early period of the disease. The use of such specific and sensitive method as the coagglutination test for this purpose accelerates and facilitates the diagnosis of typhoid fever.

[Determination of O and Vi antigens in typhoid fever in the slide coagglutination reaction]. / Opredelenie O- i Vi-antigenov pri briushnom tife v reaktsii koaggliutinatsii na stekle.

Investigations with a view to the development and trial of a slide coagglutination test system for the detection of specific Salmonella typhi antigens have been made. As a result, diagnostic agents with sensitivity to group D Salmonella lipopolysaccharides and Vi-antigen, equal to 1 X 10(-5) to 1 X 10(-6) g/l, have been obtained. Specimens of saliva, urine and fecal filtrates from 61 adult patients with bacteriologically confirmed typhoid fever and 54 practically healthy persons have been studied. The coagglutination test has been positive with specimens from 90 +/- 4% of typhoid fever patients and, within the first 5 days of the disease, with those from 85 +/- 7% of such patients. The slide coagglutination test with saliva specimens has been found to be more informative than that with urine specimens. The data obtained in these investigations indicate that the slide coagglutination test is highly sensitive and specific, which offers good prospects for its use as a simple, economic and demonstrative method for the early tentative diagnosis of typhoid fever.