Evolution of Helicobacter pylori susceptibility to antibiotics during a 10-year period in Lithuania.

The study evaluated the changes in the prevalence of Helicobacter pylori strains with primary resistance to antibiotics during the last 10 years in Lithuania. H. pylori susceptibilities to antibiotics were tested in 89 patients in 1998, in 81 patients in 2001 and in 90 patients in 2007/2008. Susceptibility to metronidazole, clarithromycin, amoxicillin and tetracycline was tested using E-test or agar dilution method. Susceptibility to ciprofloxacin was only tested in 2007/2008. Data about utilization of all authorized and available on market macrolides and clindamycin in Lithuania during 2003-2007 were evaluated using WHO ATC/DDD methodology. A total of 260 H. pylori strains cultured from untreated adult patients were investigated. Primary resistance rates (1998, 2001 and 2007/2008) for metronidazole were 24.7%, 33.3%, and 35.6%, for clarithromycin 1.1%, 3.7%, and 3.3% and for tetracycline 0%, 2.5% and 0% respectively. No cases of amoxicillin resistance have been detected. The resistance rate for ciprofloxacin was 5.6% in 2007/2008. Data of total macrolides and clarithromycin utilization in Lithuania revealed that despite an increase of consumption of these drugs in Lithuania during 2003-2007 in 1.5 times, the total macrolide consumption remains one of the lowest in Europe. We have not observed any significant changes in the susceptibility of H. pylori to the most widely used antibiotics during the recent 10-year period. The low resistance rate to clarithromycin might be related to the policy to avoid use of macrolides as first-line treatment for pulmonary and other infections.

CD4+CD25(high) Treg cells in peripheral blood during remission and exacerbation of allergic asthma in children.

AIM: To determine the percentage of CD4+CD25(high) Treg cells in peripheral bloodCD4+ T cells of allergic asthmatic children during disease remission and exacerbation. METHODS: Peripheral blood mononuclear cells (PBMC) and serum samples were collected from 6- to 11-year-old children with mild-to-moderate allergic asthma (n = 34)and from healthy controls (n = 15). CD4+CD25(high) T cells in PBMC were detected by flow cytometry. Total and specific IgE in serum were analysed by enzyme-amplified chemiluminescence, and IL-2 was measured by ELISA. RESULTS: There was no significant difference in CD4+CD25(high) T-cell proportions between asthmatic children in exacerbation and remission as compared with controls.CD4+CD25(high) T-cell percentages were not correlated with total and specific IgE. IL-2 was elevated in both disease remission and exacerbation but did not correlate significantly with CD4+CD25(high) T-cell percentages. CONCLUSION: CD4+CD25(high) T-cell proportion in the peripheral blood of total CD4+T cells is not reduced in children with allergic IgE-mediated asthma and does not differ between disease remission and exacerbation.

Helicobacter pylori genotypes in Lithuanian patients with chronic gastritis and duodenal ulcer.

OBJECTIVE: Clinical outcome of Helicobacter pylori (H. pylori) infection might be associated with specific virulence-associated bacterial genotypes. The distribution of different bacterial genotypes varies geographically. The aim of this study was to assess the relationship between cagPAI, vacA, and iceA status and severity of the disease in patients from Lithuania, infected by H. pylori. MATERIAL AND METHODS: H. pylori from 81 patients (37 with duodenal ulcer and 44 with chronic gastritis) was isolated from gastric biopsy specimens and cultured. Bacterial genotypes cagPAI, vacA (s and m subtypes) and iceA were analyzed by polymerase chain reaction using specific primers. RESULTS: The cagPAI was identified in 59.3% of Lithuanian H. pylori strains investigated. H. pylori strains cultured from duodenal ulcer (DU) patients more frequently (P<0.01) contained cagPAI and vacA s1 genotypes (75.7% and 75.7%, respectively) in comparison to isolates from chronic gastritis (CG) patients (45.5% and 40.9%, respectively). Evaluation of nucleotide sequence of the vacA middle-region revealed that vacA s2/m2 genotype was more frequent in CG than in DU patients (56.8% and 24.3%, respectively; P<0.05). We have not found any differences in the frequency of iceA1 genotype between the DU and CG patients (46.0% and 40.9%, respectively; P>0.05). CONCLUSION: Our study suggests that cagPAI and vacA s1 genotypes are associated with peptic ulceration in Lithuanian patients infected by H. pylori.

Immune system alterations in patients with inflammatory bowel disease during remission.

OBJECTIVE: Perturbed immune homeostasis elicited by misbalanced production of proinflammatory and anti-inflammatory cytokines is characteristic of inflammatory bowel disease. The aim of this study was to evaluate cytokine profile in patients with different forms of inflammatory bowel disease - ulcerative colitis and Crohn's disease - during clinical remission phase. MATERIAL AND METHODS: Production of proinflammatory Th1 cytokines (tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma)) and anti-inflammatory Th2 cytokines (interleukin-10 (IL-10) and interleukin-13 (IL-13)) was analyzed in peripheral blood mononuclear cells of patients with inflammatory bowel disease (9 with ulcerative colitis and 9 with Crohn's disease) and control subjects (n=11) by enzyme-linked immunosorbent assay (two-site ELISA). RESULTS: The results of the study revealed that the level of TNF-alpha after stimulation with phytohemagglutinin in patients with Crohn's disease was significantly higher in comparison to both patients with ulcerative colitis and controls (P<0.001 and P<0.01, respectively). The secretion of IFN-gamma both in patients with Crohn's disease and ulcerative colitis was lower than that in controls (P=0.05 and P<0.01, respectively), but it normalized after stimulation with phytohemagglutinin. The levels of IL-10 and IL-13 were significantly (P<0.01) higher in patients with Crohn's disease than in patients with ulcerative colitis and control group before and after stimulation with phytohemagglutinin. CONCLUSIONS: The results of our study provide evidence that in patients with inflammatory bowel disease, the imbalance between production of proinflammatory Th1 and anti-inflammatory Th2 cytokines persists even during remission of the disease, and disturbances of immune homeostasis are significantly more expressed in patients with Crohn's disease than in patients with ulcerative colitis.

Soluble HLA-G in the peritoneal fluid of women with endometriosis.

The aim of this study was to determine the level of soluble HLA-G molecules in the peritoneal fluid of endometriosis patients. The findings demonstrate that a soluble HLA-G level in the peritoneal fluid of women with endometriosis is similar to that of the control group.

Regulation of epsilon germline transcription and switch region mutations by IgH locus 3' enhancers in transgenic mice.

Germline (GL) transcription is regulated by specific promoters and immunoglobulin heavy chain (IgH) 3' locus enhancers and is necessary for Ig class-switch recombination (CSR). We have generated different transgenic lines containing the GL epsilon promoter, switch (S) epsilon region, and constant (C) epsilon region with or without the DNase I-sensitive regions (HS) 3A-HS1,2 or HS3B-HS4 3' IgH enhancer pairs. The enhancerless construct was expressed in B cells activated by interleukin (IL)-4 and CD40, thus resembling regulation of the endogenous gene. Both enhancer-containing transgenes efficiently increased expression in B cells and were strongly up-regulated by stimuli. In addition, Sepsilon regions of the transgene containing HS3B-HS4 were mutated in activated, sorted B cells. Such mutations are known to precede CSR and are dependent on activation-induced cytidine deaminase (AID). Our findings show that all elements necessary for recruitment of the recombination machinery are present in the transgene containing HS3 and HS4. These enhancers probably provide something more specific than mere increased accessibility of switch regions. We propose that transcription factors binding the enhancers help to target the recombination machinery to the switch regions.

Aberrant expression of CD95 and CD69 molecules among CD56 cells in women with endometriosis.

PROBLEM: Activated lymphocytes can be eliminated by Fas/Fas ligand (FasL)-induced cell death. Endometrial cells express FasL. The aim of our study was to determine the expression of CD56+ cells (natural killer and natural killer T cells) Fas antigen CD95 and the early activation molecule CD69 in the peritoneal fluid of women with endometriosis. METHOD: Two-colour flow cytometry was used. RESULTS: In the early stages of endometriosis, more CD56+ cells expressed CD69 and CD95 molecules when compared with the control group. However, in case of severe endometriosis the percentage of CD95+CD56+ cells in peritoneal fluid was similar to that of the control group, but the expression of CD69 molecules remained high. CONCLUSION: Because of Fas/FasL mechanisms, in the initial stages of endometriosis the activated peritoneal fluid CD56+ cells can be intensively eliminated, thus providing conditions for the survival of ectopic endometrial cells and the development of the disease.

Endometrial and peritoneal macrophages: expression of activation and adhesion molecules.

PROBLEM: Macrophages are highly individualized in tissues and their activities are a reflection of systemic and local environmental signals. The expression of activation (CD69, CD71) and adhesion (CD54) molecules on the surface of CD14+ endometrial macrophages at various phases of the menstrual cycle was compared with the cell surface receptors of peritoneal fluid macrophages. METHOD OF STUDY: Two-colour-flow cytometry was used to determine the peritoneal and endometrial macrophage phenotype. RESULTS: Endometrium macrophages expressed a lower level of CD69+ and CD54+ macrophages than peritoneal macrophages. However, CD71 receptors displayed similar expression in both macrophage populations, endometrium and peritoneal, except during the proliferative phase. CONCLUSION: These findings demonstrate the differences between macrophages from endometrium and peritoneal fluid with regard to CD69, CD71 and CD54 expression. In addition, increased numbers of endometrial macrophages in the late secretory phase of the menstrual cycle suggest that they may play a role in menstruation.

Activity of eosinophils and immunoglobulin E concentration in the peritoneal fluid of women with endometriosis.

Autoinflammatory phenomena, including autoantibody production and atopy, have been regarded as associated with endometriosis. The present study investigates the activity of eosinophils and the distribution of immunoglobulin E concentrations in the peritoneal fluid of women with early endometriosis. The study group consisted of 30 patients with laparoscopically diagnosed early endometriosis. The healthy control group consisted of 18 females with no evident changes in the abdominal cavity and no endometrial foci. Concentrations of immunoglobulin E in serum and peritoneal blood were determined by enzyme immunoassay. The activity of eosinophils was estimated according to the expression of the early activation molecule CD69 by the flow cytometry method. The concentrations of immunoglobulin E in the peripheral blood and peritoneal fluid were similar in both groups. However, the count of CD69+ eosinophils was higher in the peritoneal fluid of women with endometriosis. The results indicate that activated eosinophils accumulate in the peritoneal fluid in early endometriosis and can play a significant role in the pathogenesis of the disease.

Differential expression of KIR/NKAT2 and CD94 molecules on decidual and peripheral blood CD56bright and CD56dim natural killer cell subsets.

OBJECTIVE: To investigate killer inhibitory receptor (KIR) expression by natural killer (NK) cells in early pregnancy. DESIGN: Case-control study of immunologic markers. SETTING: University hospital. PATIENT(S): Thirty pregnant women and 22 nonpregnant women. INTERVENTION(S): Peripheral venous blood sampling and decidual tissue collection after elective abortion. MAIN OUTCOME MEASURE(S): Flow cytometry was used to assess expression of KIR by NK cells in the cell samples. RESULT(S): In contrast to CD56(bright) peripheral blood NK cells, CD56(dim) cells express killer cell Ig-like receptor KIR/NKAT2. However, KIR/NKAT2 and lectin-like CD94 are present on both subsets of decidual NK cells. We found no differences between peripheral blood NK cell subsets from pregnant and nonpregnant women. CONCLUSION(S): Our findings demonstrate that NK cell subsets, distributed in accordance with CD56 molecule density on cell surface, express killer inhibitory receptors CD94 and KIR/NKAT2 in a different way. Our data support the view that CD56(bright)KIR/NKAT2+CD94+ decidual NK cells are specialized NK cells that have an important role to play in early pregnancy.