[Changes in blood CD4+CD25+ regulatory T cells in children with severe purulent meningitis].

OBJECTIVE: To preliminarily study the changes in CD4+CD25+ regulatory T cells (Tregs) in children with severe purulent meningitis at the early stage and its possible implications. METHODS: A retrospective analysis was performed on the clinical data of 39 children with severe purulent meningitis who were admitted to the pediatric intensive care unit from August 2014 to December 2015. According to whether Tregs count was decreased within 12 hours of hospitalization (considering Tregs count <410/mm3 as decreased), they were divided into two groups: decrease group and non-decrease group. The associations between the changes in Tregs cells and the clinical manifestations, laboratory marker levels, and prognosis were analyzed. RESULTS: Of the 39 cases, 13 (33%) showed a decrease in the proportion of Tregs cells (<31%) and 18 (46%) showed a decrease in the absolute Tregs cell count (<410/mm3). Four deaths were all in the Tregs decrease group. Compared with the non-decrease group, the decrease group showed a significantly higher proportion of children with a peripheral blood leukocyte count lower than the normal range and a significantly greater increase in the level of serum procalcitonin (P<0.05). CONCLUSIONS: Tregs might be suppressed in children with severe purulent meningitis at the early stage. And its suppression could be related to the severer inflammation reaction and higher mortality in those patients.

[CLINIC-EXPERIMENTAL SUBSTANTIATION OF OZONOTHERAPY APPLICATION IN THE TREATMENT OF PURULENT CHOLANGITIS AND OBTURATION JAUNDICE].

Efficacy of the ozonotherapy application as an important component of complex treatment in purulent cholangitis (PCH) was studied. In choledocholithiasis (without infectioning of bile) ozonotherapy may be prescribed as additional component at complex treatment. In PCH ozonotherapy application have promoted the improvement of laboratory indices in 6.6 times, comparing with such, occurring after basic therapy.

Entamoeba histolytica antigenic protein detected in pus aspirates from patients with amoebic liver abscess.

Entamoeba histolytica is a causative agent of amoebic liver abscess (ALA) and is endemic in many underdeveloped countries. We investigated antigenic E. histolytica proteins in liver abscess aspirates using proteomics approach. Pus samples were first tested by real-time PCR to confirm the presence of E. histolytica DNA and the corresponding serum samples tested for E. histolytica-specific IgG by a commercial ELISA. Proteins were extracted from three and one pool(s) of pus samples from ALA and PLA (pyogenic liver abscess) patients respectively, followed by analysis using isoelectric focussing, SDS-PAGE and Western blot. Unpurified pooled serum samples from infected hamsters and pooled human amoebic-specific IgG were used as primary antibodies. The antigenic protein band was excised from the gel, digested and analysed by MALDI-TOF/TOF and LC-MS/MS. The results using both primary antibodies showed an antigenic protein band of ∼14kDa. Based on the mass spectrum analysis, putative tyrosine kinase is the most probable identification of the antigenic band.

[The immune activity of cytoflavine in the treatment of the diffuse septic peritonitis].

Influence of citoflavin on dynamics of functional condition of mitogen-induced immunocompetent blood cells at patients with widespread purulent peritonitis is studied. It is established that investigated immunocompetent cells have regulating effect on migration of neutrophil leukocytes at widespread purulent peritonitis during all postoperative period. PHA-activated mononuclear cells don't cause inhibition of migration of neutrophil granulocytes, observed in norm. At the same time the LPS-activated monocytes lead to migration strengthening of neutrophil granulocytes, in comparison with the data received in norm. Usage of citoflavin at patients with widespread purulent peritonitis renders corrective action on immunocompetent cells, changing their properties of regulation of migratory activity of neutrophil granulocytes. It is revealed that such action of citoflavin is connected with function restoration of mononuclear cells to inhibit migration of neutrophil leukocytes and with restoration of stimulation of migration of neutrophil leukocytes under the influence of the LPS-activated monocytes to the values received in norm. The revealed effects at use of citoflavin are most expressed by the 5th day of the postoperative period.

[Clinical efficacy of immunostimulating drugs in treatment of purulent infections].

81 patients with the acute soft tissue purulent infections were divided in 4 groups and treated with the use of traditional means without immunostimulating drugs; with immunoglobulins; with derinate and combination of both, respectively. The last treatment modality demonstrated maximal clinical efficacy. That is the decrease of the wound contamination period and inflammatory process; acceleration of granulation and epithelization. Clinical recovery was accompanied by marked positive changes of cell and humoral immune status.

Systemic Th17-like cytokine pattern in gangrenous appendicitis but not in phlegmonous appendicitis.

BACKGROUND: Increasing circumstantial evidence suggests that not all patients with appendicitis will progress to perforation and that appendicitis that resolves may be a common event. Based on this theory and on indications of aberrant regulation of inflammation in gangrenous appendicitis, we hypothesized that phlegmonous and gangrenous appendicitis are different entities with divergent immunoregulation. METHODS: Blood samples were collected from patients with gangrenous appendicitis (n = 16), phlegmonous appendicitis (n = 21), and nonspecific abdominal pain (n = 42). Using multiplex bead arrays, we analyzed a range of inflammatory markers, such as interleukin (IL)-1ra, IL-1rbeta, IL-2, IL-6, IL-10, IL-12p70, IL-15, and IL-17; interferon-gamma; tumor necrosis factor; CXCL8; CCL2; CCL3; and matrix metalloproteinase (MMP)-1 MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-12, and MMP-13 in blood. RESULTS: Compared with patients with phlegmonous appendicitis and nonspecific abdominal pain, the patients with gangrenous appendicitis had increased levels of the proinflammatory markers IL-6, CCL2, IL-17, MMP-8, and MMP-9 (P < or = .04 each) accompanied by increased levels of the anti-inflammatory cytokines IL-1ra and IL-10 (P < or = .02). Patients with phlegmonous appendicitis had increased levels of IL-10 only. CONCLUSION: The finding of a pattern of inflammatory markers compatible with the highly inflammatory Th17 subset in sera from patients with gangrenous appendicitis, but not in phlegmonous appendicitis, supports the hypothesis that gangrenous and phlegmonous appendicitis are different entities with divergent immune regulation. Additional studies of the differential immunopathogenesis of phlegmonous and gangrenous appendicitis are warranted, as this may have important implications in the diagnosis and management of patients with suspicion of appendicitis.

[Microflora dynamics in purulent skin wound in rats after ozonized perftorane applications].

In 30 Wistar rats divided in 3 groups with 10 rats in each skin defects were reproduced under anaesthesia and wound surface was fixed with the help of plastic rings. The wounds were contaminated with microbe dredge consisting of St. aureus, Ps. aeruginosa and E. coli in the concentration of 10(9) cells/ml. After 24, 48, 72, 96 and 120 hours the wound was irrigated with 10-15 ml of the following solutions: in the 1st group - commercial perftorane solution, in the 2nd group - ozonized perftorane, in the 3rd group - physiological solution. Hard medium inoculation let determine total number of growing microbe colonies. The material was taken each 3rd day. 9 experimental series were executed with the total length of experiment equal to 30 days. It was established that from 15th day of the experiment in the group with ozonized perftorane irrigations the mean number of microorganisms in the wound was reduced, granulation tissue growth and epithelization were speeded up if compared with the groups of animals irrigated with physiological solution and non-ozonized perftorane. Ozone action was connected not only with its bactericidal properties but in considerable degree with comprehensive local immunity activation.

Immune defense of rats immunized with fennel honey, propolis, and bee venom against induced staphylococcal infection.

The objective of this work was to evaluate the potency of bee product-immunized rats to overcome an induced Staphylococcus aureus infection. Forty rats were divided to eight groups: T1, T3, and T5 received, respectively, fennel honey, ethanol, and aqueous propolis extracts orally, and T2, T4, and T6 were administered the respective materials intraperitoneally; T7 received bee venom by the bee sting technique; and T8 was the control group. All groups were challenged by a bovine clinical mastitis isolate of S. aureus. Each rat received 2 mL of broth inoculated with 1 x 10(5) colony-forming units/mL intraperitoneally. Two weeks post-induced infection all rats were sacrificed and eviscerated for postmortem inspection and histopathological study. Three rats from T8 and one rat from T7 died before sacrifice. Another two rats, one each in T4 and T5, had morbidity manifestations. The remaining experimental animals showed apparently healthy conditions until time of sacrifice. Postmortem inspection revealed that all T8 rats showed different degrees of skeletal muscle and internal organ paleness with scattered focal pus nodules mainly on lungs and livers. All rats of the treated groups showed normal postmortem features except three rats. A dead rat in group T7 showed focal pus nodules on the lung surface only, whereas the affected two rats in groups T4 and T5 appeared normal except with some pus nodules, but much smaller than in the control, scattered on the hepatic surface and mesentery. Histopathological studies revealed that T8 rats had typical suppurative bronchopneumonia and or severe degenerative and necrobiotic changes in hepatic tissues. Three affected rats of the treated groups showed slight bronchopneumonia or degenerative hepatic changes only. The other animals of the treated groups showed completely normal parenchymatous organs with stimulated lymphatic tissues. It was concluded that all tested previously bee product-immunized rats could significantly challenge the induced S. aureus infection (P < .01). The effects were more pronounced in rats that had received fennel honey solution.

Suppurative inflammation with microabscess and pseudocyst formation is a characteristic histologic manifestation of cutaneous infections with rapid-growing Mycobacterium species.

Mycobacterial infections of the skin classically cause a granulomatous tissue reaction. We have observed a suppurative pattern of inflammation associated with infections by rapid-growing Mycobacterium species in immunocompromised patients. We report 6 cases in skin and soft tissue with an unusual but consistent lack of a predominance of granulomatous inflammation. Of the 6 cases, 4 had predominantly (approximately 75%) suppurative inflammation, 1 case predominantly demonstrated (approximately 75%) a mix of acute and chronic inflammation, and 1 case showed an approximately equal contribution of suppurative and granulomatous inflammation. All 6 cases showed abscess formation and numerous acid-fast bacilli (AFB) on AFB stain and were confirmed by tissue culture. Of these 6 cases, 2 had microabscesses with central pseudocysts harboring microorganisms. Five patients were taking oral prednisone, and 1 had an uncharacterized immunodeficiency. These cases highlight the need for awareness of this unusual manifestation of infection with rapid-growing Mycobacterium species, particularly in immunocompromised patients.

Staphylococcus and the healing power of pus.

Staphylococcus aureus infections induce formation of neutrophil-rich abscesses filled with debris from dead phagocytes. Corbin and colleagues report that this pus has antimicrobial powers through the activity of calprotectin. Calprotectin, a member of the S100 family of proinflammatory proteins, acts through chelation of manganese. As manganese is an essential cofactor for several enzymes in S. aureus, this impacts bacterial growth and the bacterium's ability to withstand oxidative stress.

Detection of excretory Entamoeba histolytica DNA in the urine, and detection of E. histolytica DNA and lectin antigen in the liver abscess pus for the diagnosis of amoebic liver abscess.

BACKGROUND: Amoebic liver abscess (ALA) and pyogenic liver abscesses (PLA) appear identical by ultrasound and other imaging techniques. Collection of blood or liver abscess pus for diagnosis of liver abscesses is an invasive procedure, and the procedure requires technical expertise and disposable syringes. Collection of urine is a noninvasive procedure. Therefore, there has been much interest shown towards the use of urine as an alternative clinical specimen for the diagnosis of some parasitic infections. Here, we report for the first time the detection of E. histolytica DNA excreted in the urine for diagnosis of the cases of ALA. RESULTS: E. histolytica DNA was detected in liver abscess pus specimen of 80.4% of ALA patients by a nested multiplex polymerase chain reaction (PCR) targeting 16S-like r RNA gene. The nested PCR detected E. histolytica DNA in all 37 (100%) liver abscess pus specimens collected prior to metronidazole treatment, but were detected in only 53 of 75 (70.6%) pus specimens collected after therapy with metronidazole. Similarly, the PCR detected E. histolytica DNA in 21 of 53 (39.6%) urine specimens of ALA patients. The test detected E. histolytica DNA in only 4 of 23 (17.4%) urine specimens collected prior to metronidazole treatment, but were detected in 17 of 30 (56.7%) urine specimens collected after treatment with metronidazole. The enzyme-linked immunosorbent assay (ELISA) for the detection of lectin E. histolytica antigen in the liver abscess pus showed a sensitivity of 50% and the indirect haemagglutination (IHA) test for detection of amoebic antibodies in the serum showed a sensitivity of 76.8% for the diagnosis of the ALA. CONCLUSION: The present study for the first time shows that the kidney barrier in ALA patients is permeable to E. histolytica DNA molecule resulting in excretion of E. histolytica DNA in urine which can be detected by PCR. The study also shows that the PCR for detection of E. histolytica DNA in urine of patients with ALA can also be used as a prognostic marker to assess the course of the diseases following therapy by metronidazole. The detection of E. histolytica DNA in urine specimen of ALA patients provides a new approach for the diagnosis of ALA.

[The role of ozone therapy in prevention of pyoinflammatory complications after transurethral resection of prostatic adenoma].

The study of efficacy of ozone therapy (OT) in preparation of patients with prostatic adenoma (PA) and cystostoma for transurethral resection (TUR) of PA included 20 PA patients with cystostomic drainage who had undergone PA TUR and preoperative preparation with OT sessions. The control group consisted of similar patients but without ozone pretreatment. OT efficacy was assessed by the rate of pyoinflammatory complications (PIC), results of immunological examination, positive changes in prostatic secretion, urine analysis, total blood count, degree ofbacteriuria. In the study group PIC (acute urethritis) developed in 1 patient, in the control--in 6 patients (3 cases of acute urethritis, 2 cases of acute prostatitis and 1 case of acute epididymitis). OT led to lowering of mean values of leukocyturia from 18.1 +/- 0.6 to 14.3 +/- 0.7 on the day of operation, to 10.9 +/- 0.7 after 4 days and to 8.7 +/- 0.6 on postoperative day 8 versus from 18.8 +/- 0.8 to 15.4 +/- 0.7, to 15.8 +/- 0.6 and 13.5 +/- 0.6, respectively. Mean control count of leukocytes in prostatic secretion fell in both groups. OT reduced bacteriuria. Number of bacteria to the day of operation decreased in both groups, but complete elimination of the agent from urine on day 8 was not achieved in the controls. The study group exhibited a rise in the absolute count of blood leukocytes, lymphocytes and ESR diminishing. OT raised significantly the phagocytic count and activity, concentration of mature T-lymphocytes (CD3), T-helpers (CD4), cytotoxic T-lymphocytes (CD8), B-lymphocytes (CD20), T-NK-cells (CD16), T-lymphocyte activation markers (CD3+, CD16+, CD56+, CD3+, CD25+, HLADR+, CD3+. The concentration of IgG, IgM, IgA remained high. Thus, the results of ozone application before PA TUR appeared promising in prevention of postoperative PIC.

[Immunocorrectors in the complex treatment of postoperative suppurative-inflammatory complications in surgical patients and monitoring of immunological parameters]. / Immunokorrektory v kompleksnom lechenii posleoperatsionnykh gnoino-vospalitel'nykh oslozhnenii u khirurgicheskikh bol'nykh i monitoring immunologicheskikh pokazatelei.

The results of monitoring of immune parameters in different categories of surgical patients with postoperative purulent-inflammatory complications are described in the paper. The combined type of secondary immunodeficiency and degree second immunodeficiency according to A.M. Zemskov were detected in all patients. Immunocorrecting drugs, like polyoxidonium, imunofan and likopid, were added to the complex treatment of patients with the above complications. Their administration normalized the dynamic clinical course of the purulent-inflammatory processes and restored the immunological parameters.

Effect of ovarian hormones on periodical changes in immune resistance associated with estrous cycle in the beagle bitch.

In bitches, the onset of pyometra, an infection of the uterus, characteristically occurs in the first half of the diestrous stage in the estrous cycle, in which the blood concentration of progesterone peaks and that of estradiol-17beta is lowest. To investigate the immunological mechanisms governing stage-specific onset of pyometra, peripheral blood mononuclear cells (PBMNCs) were collected from beagle bitches during different stages of the estrous cycle and examined using various immunological assays. When we examined the proliferative response of PBMNCs to PYO-252, that is a clone of Escherichia coli isolated from the uterus of a dog afflicted with pyometra, the response of PBMNCs significantly decreased in the first half (day 10) of diestrus, but increased in proestrus/estrus. No significant differences were observed in the responses to concanavaline A between stages of the cycle. Throughout the estrous cycle, canine PBMNCs did not respond to lipopolysaccharide derived from E. coli. The response of PBMNCs collected in anestrus to PYO-252 was significantly enhanced upon the addition of estradiol-17beta to the culture. In contrast, these responses were significantly suppressed in the presence of progesterone. Progesterone progenitor or metabolite molecules, which have a low affinity for the progesterone receptor, did not affect proliferative responses. Expression of gamma interferon (IFNgamma) in response to PYO-252 was also significantly enhanced by estradiol-17beta, but suppressed by progesterone. This evidence suggests that in the first half of the diestrous stage, suppressed activity of cellular immunity results from increasing progesterone concentration and minimal estrogen release. This marked decrease of immune resistance allows the expansion of E. coli, which enter the uterine cavity through the loosened cervical canal during estrus, leading to pyometra onset.

Differences in host susceptibility to disease progression in the human challenge model of Haemophilus ducreyi infection.

With human volunteers inoculated at two sites with Haemophilus ducreyi, outcomes for a subject were not independent. In a reinfection trial, 2 of 11 previous pustule formers and 6 of 10 previous resolvers resolved all sites of infection. There was no correlation between serum bactericidal or phagocytic activity and outcome in the trial. These data indicate that different hosts are differentially susceptible to disease progression versus resolution in the model.

[Diagnosis and differentiated treatment of secondary immunodeficiencies]. / Diagnostika i differentsirovannoe lechenie vtorichnykh immunodefitsitov.

AIM: To evaluate alterations in the immune system (IS) in patients with different forms of secondary immunodeficiency and design of differentiated programs of reestablishment of defective functions depending on pathogenetically important type of deficiency of immunocompetent cells. MATERIALS AND METHODS: Clinicoimmunological examination was made in 678 patients with complicated course of infectious-inflammatory diseases. Immunotropic medicines and physicochemical impacts were used in accordance with types of disorders in the system of immune homeostasis. RESULTS: There was a pathogenetic heterogeneity of IS disorders in complicated course of infectious-inflammatory diseases: generalized forms of infection (bacterial shock, sepsis) are in 75% of cases associated with deficiency of effector functions of peripheral blood polymorphonuclear leukocytes of the second-third degree, progressive fall in production of IgG immunoglobulins (42%), cellular-humoral immunodeficiency (92%). In lingering acute inflammatory diseases activation of phagocytosis occurred in 30%, IgG and/or IgM rise was in 50%, phagocytic function deficiency occurred in 48%, low production of immunoglobulins in 24%, humoral-cellular immunodeficiency in 62%. Purulent infection is associated with secondary cellular-humoral immunodeficiency, lowering of the immunoregulatory index (47%), phagocytic function deficiency (up to 35%), hyperproduction of IgM. Recurrent bacterial-viral diseases form in immunocompromised patients with T-lymphocytopenia (56%) and cellular-humoral immunodeficiency (30%). CONCLUSION: Protracted chronic inflammatory diseases are characterized by variability of changes in the immune systems. Combined types of disorders were found in 52% of the examinees. Pathogenetic heterogeneity of the disorders are determined by concomitant and previous diseases, occupational hazards and intoxication, environmental conditions, etc. CONCLUSION: Immunocorrective therapy in secondary immunodeficiency is conducted with allowances for pathogenetically essential types of disorders in the system of immune homeostasis, clinical variant of complication of inflammatory process under control of immunogram.