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Biomedical subjects

U Demkow

Publications and source records attributed to U Demkow.

At least 19 recordsLinked to original sources
PubMed2006

Humoral immune response against mycobacterial antigens in children with tuberculosis.

Different clinical outcomes of tuberculosis (TB) are related to the balance between cell-mediated and humoral immunity and may depend on environmental and individual factors including age and sex. The purpose of this study was to analyze the humoral immune response to recombinant and native mycobacterial antigens in relation to clinical presentations of pulmonary TB in children. We examined 224 serum samples including 81 primary and 31 postprimary TB cases, 30 cases of latent TB infection, and 82 nontuberculosis controls. Commercially available ELISA assays detecting IgG, IgA, and IgM against antigens: 38 kDa, 16 kDa, 38 kDa, lipoarabinomannan (LAM), and A-60 were used. The results indicate that IgG production was very low in primary compared with postprimary TB (P<0.0001). IgM levels did not differ between the examined groups. Antibody levels strongly depended on the child's age. In infants aged below 1 there was no difference in the antibody level between the TB and control cases. Most positive cases were observed in children aged above 10. The influence of BCG vaccination on the antibody level was not seen. In all subgroups, person-to-person heterogeneity of antigen recognition was observed. We conclude that humoral immune response is associated with the phase of TB and is stronger in more advanced TB forms. IgG and IgA production against mycobacterial antigens is very low in young children.

Adolescent↗
PubMed2006

Kinetics of calcium ion concentration accompanying signal transduction in neutrophils from children with increased susceptibility to infections.

Increased susceptibility to infections can be a consequence of altered function of immune cells including neutrophils. The goal of the study was to evaluate the process of neutrophil activation via Ca(2+)-mediated signal. The study was performed on isolated peripheral blood neutrophils obtained from 41 children with recurrent infections (21 girls, 20 boys) 3-10 years old with more than five episodes of respiratory tract infection (RI) per year and from a control group of 30 healthy children age and sex matched, free from allergic, immune and hematological disorders. Neutrophils were activated by bacterial peptide fMLP, opsonized zymosan (OZ), and phorbol myristat acetate (PMA). The kinetics of the intracellular Ca(2+) concentration i[Ca(2+) ] was assessed by flow cytometry (Coulter Epics XL) with the use of Fluo3 and Fura Red fluorescent dyes. Data were collected in histograms displaying Fluo3 fluorescence vs. time and Fura Red fluorescence vs. time and the mean channels of fluorescence intensity were used for calculations. fMLP and OZ-induced Ca(2+) mobilization lasted shorter in the RI group (P<0.05). The peak influx of free Ca(2+) and i[Ca(2+) ] in the resting state after stimulation with fMLP were lower in the patients (P<0.05). In the RI group stimulation with OZ was delayed compared with that in the control group (P<0.01). In response to PMA, i[Ca(2+) ] decreased faster. The kinetic slopes of i[Ca(2+) ] in both groups examined differed statistically at all points measured. A decrease in i[Ca(2+) ] after PMA stimulation was greater (P<0.01) and lasted longer in the RI group. We conclude that increased sensitivity to infections in RI children may be related to the disturbance in neutrophil activation that is mediated by changes in i[Ca(2+) ] with the subsequent production of free oxygen radicals. Such a disturbance may be inheritable or secondary to infection and antibiotic therapy.

Adolescent↗
PubMed2006

Differences in responses upon corticosteroid therapy between smoking and non-smoking patients with COPD.

Inhaled corticosteroids have a high level of topical anti-inflammatory activity. However, in patients with COPD these drugs have been reported to exert limited effects. A reduction in histone deacetylase (HDAC) activity is suggested to prevent the anti-inflammatory action of corticosteroids. Cigarette smoke is known to reduce HDAC expression. The aim of this study is to compare the outcome of corticosteroid therapy in both smoking and non-smoking COPD patients. Twenty-three smoking patients and 18 ex-smoking patients with COPD were treated with inhaled corticosteroids for a period of 2 months. Blood and induced sputum samples were collected before and after treatment. Values of FEV(1) %-predicted did not change upon the therapy, but there was a trend to improve in the ex-smokers (63.1 -> 64.8%-pred.), compared with a decrease in the smokers (63.3 -> 61.6%-pred.). The levels of the pro-inflammatory cytokine IL-8 increased in the group of smokers from 379 +/-78 to 526 +/-118 ng/ml. Although not significant, a slight decrease from 382 +/-70 to 342 +/-62 ng/ml was observed in the group of ex-smokers. The neutrophil related elastase activity showed similar effects after steroid treatment, it went up from 36.4 +/-12.0 to 113.5 +/-9.7 nmol/l in smokers, and decreased from 346.2 +/-72.1 to 131.1 +/-6.5 nmol/l in ex-smokers with COPD. These results support the evidence that inhaled corticosteroids have no anti-inflammatory effects in COPD patients, but only when these patients are still smoking. Smoking cessation seems the best therapy for COPD patients.

Adrenal Cortex Hormones↗
PubMed2006

Influence of N-acetylcysteine on ICAM-1 expression and IL-8 release from endothelial and epithelial cells.

Chronic obstructive pulmonary disease (COPD) is characterized by chronic airway inflammation. The initial step in the inflammatory process is overexpression of adhesion molecules, which leads to excessive transmigration of neutrophils. One of these adhesion molecules is ICAM-1 which is elevated in COPD patients. In this study we evaluated the influence of N-acetylcysteine (NAC) (0.01 mM-30 mM) on the cytokine-induced (TNF-alpha/IL-1 beta) expression of the ICAM-1 adhesion molecule and on IL-8 release in endothelial (ECV-304) and bronchial epithelial (H292) cell lines. The methodology used consisted of immunochemistry for the assessment of surface ICAM-1 and ELISA method for that of soluble ICAM-1 and IL-8. NAC inhibited the TNF-alpha/IL-1 beta-stimulated ICAM-1 expression and IL-8 release from both cell lines in a concentration dependent manner. The most effective concentrations were 30 mM and 20 mM (99 and 90% inhibition respectively, P<0.01). We conclude that NAC is an effective inhibitor of TNF-alpha/IL-1 beta- stimulated ICAM-1 and IL-8 release in endothelial and epithelial cells. This fact highlights the anti-inflammatory potential of NAC in COPD.

Acetylcysteine↗
PubMed2006

The Gln223Arg polymorphism of the leptin receptor gene and peripheral blood/bone marrow leptin level in leukemic children.

Leptin is an adipocyte-derived hormone regulating energy homeostasis and body weight. Leptin also plays a role in hematopoiesis, cell cycle regulation, and in oncogenesis. The leptin receptor is a single transmembrane protein belonging to the superfamily of cytokine receptors, structurally related to the hemopoietin receptor family. The aim of the study was to evaluate bone marrow and peripheral blood leptin level and frequency of distribution of leptin receptor gene polymorphism Gln223Arg in children with acute leukemia. The examined group included 92 children with acute leukemia (83 ALL and 9 AML) and 39 non-leukemic control children. Leptin level was measured by ELISA method at the day of leukemia diagnosis. Genomic DNA was isolated with the use of a column method and the genotyping of DNA sequence variation was carried out by the restriction enzyme analysis of PCR - amplified DNA. The samples were then electrophoresed on 2.5% agarose gel. Leptin level in leukemic children was lower than in healthy children. Bone marrow leptin level was significantly lower than that in the blood in leukemic children with ALL-T and AML. An analysis of frequency distribution of the Gln233Arg polymorphism in the leptin receptor gene in leukemic children showed lack of differences between the patients and controls. There was no difference in the genotype frequencies between the leukemic AML and ALL groups either. The results indicate a possible relation between the leptin level and leukemia development in children. The effectory effect of the hormone seems not related to Gln223Arg polymorphism of its receptor.

Adolescent↗
PubMed2005

Humoral immune response against mycobacterial antigens in bronchoalveolar fluid from tuberculosis patients.

Resistance to tuberculosis (TB) is cell-mediated but a humoral response is common and may be correlated with the lack of effective local cellular defense mechanisms. The goal of the study was to evaluate IgG, IgA, and IgM-mediated humoral immune response against 38-kDa+16-kDa and 38-kDa+lipoarabinomannan (LAM) mycobacterial antigens in bronchoalveolar fluid (BALF) from patients with pulmonary TB. Non-tuberculosis (NTB) patients were used as control. 179 BALF samples (56 TB and 123 NTB) were examined. Commercially available ELISA-based assays against proteins 38-kDa and 16-kDa or 38-kDa plus LAM were used. Three different dilutions of BALF: 1:1; 1:10, and 1:50 (100) were tested. Only the results obtained with the 1:10 dilution allowed distinguishing TB and NTB groups. The mean IgG level for 38-Da+LAM was significantly higher in the TB than that in the NTB group (P<0.0001). The mean IgA level for 38-kDa+LAM also was higher in the TB group (P<0.05). No difference was observed between TB and NTB groups in the titer of IgM antibodies. These findings indicate that TB is associated with the presence of detectable levels of antibodies in BALF. The antibody response is highly heterogeneous. This phenomenon results from the balance between pathogen and host immune system. The tests examined for detection of IgG in BALF can be used in combination with other diagnostic methods to increase diagnostic accuracy of pulmonary TB.

Antibodies, Bacterial↗
PubMed2005

New developments in the treatment of COPD: comparing the effects of inhaled corticosteroids and N-acetylcysteine.

Inhaled corticosteroids (ICS) are widely used for the treatment of COPD despite of controversial statements concerning their efficacy. The use of N-acetylcysteine (NAC), a mucolytic drug with antioxidant properties, is less clear, but it may counteract the oxidant-antioxidant imbalance in COPD. The aim of this study was to evaluate whether treatment of COPD patients with ICS or NAC is able to improve inflammatory indices and to enhance lung function. ICS treatment enhanced protective markers for oxidative stress such as glutathione peroxidase (GPx) (51.2 +/-5.8 vs. 62.2 +/-8.6 U/g Hb, P<0.02) and trolox-equivalent antioxidant capacity (TEAC) (1.44 +/-0.05 vs. 1.52 +/-0.06 mM, P<0.05). NAC decreased sputum eosinophil cationic protein (318 +/-73 vs. 163 +/-30 ng/ml, P<0.01) and sputum IL-8 (429 +/-80 vs. 347 +/-70 ng/ml, P<0.05). The increased antioxidant capacity prevented an up-regulation of adhesion molecules, since the levels of intracellular adhesion molecule 1 (ICAM-1) correlated negatively with GPx (P<0.0001) and TEAC (P<0.0001). On the other hand, expression of adhesion molecules was promoted by inflammation, reflected by a positive correlation between the levels of IL-8 and ICAM-1 (P<0.0001). The effects of treatment on lung function were only reflected in the FEV(1) values. The absolute value of FEV(1), both before and after salbutamol inhalation, increased from 1690 +/-98 to 1764 +/-110 ml, and 1818 +/-106 to 1906 +/-116 ml, respectively, after ICS (P<0.05) . Ten weeks after treatment, FEV(1) values dropped to 1716 +/-120 ml post-salbutamol (P<0.05). When followed by treatment with NAC, these values decreased even further to 1666 +/-84 ml. These results suggest that ICS improved lung function in COPD patients with moderate airflow obstruction, beside a minor improvement in the oxidant-antioxidant imbalance leading to a lesser expression of ICAM-1. Treatment with NAC decreased some inflammatory parameters and had indirectly an inhibitory effect on the expression of adhesion molecules.

Acetylcysteine↗
PubMed2005

Effects of homogenization of induced sputum by dithiothreitol on polymorphonuclear cells.

Induced sputum represents a useful and non-invasive tool to isolate different cells from the airways. Complete homogenization of sputum is important for dispersion of cells and is usually achieved by use of dithiothreitol (DTT). However, it is not known if DTT will influence the viability and functionality of cells obtained by induced sputum. In the present study, induced sputum was processed by DTT or by PBS treatment. The obtained neutrophils were compared with neutrophils obtained from peripheral blood and from bronchoalveolar lavage fluid (BAL). These isolated neutrophils were treated in a similar way as the sputum neutrophils with DTT or PBS. All isolated cells were used for chemiluminescence tests and for the measurement of elastase and myeloperoxidase release after stimulation with fMLP. The results showed that the maximum chemiluminescence response was always significantly lower after DTT treatment: blood, 16.68 +/-1.89 vs. 2.62 +/-0.43 mV, P<0.0001; sputum, 2.96 +/-0.30 vs. 1.09 +/-0.01 mV, P<0.01; BAL, 25.47 +/-0.88 vs. 8.22+/-0.20 mV, P<0.0001. Both spontaneous and fMLP-induced release of elastase and myeloperoxidase (MPO) was in most cases enhanced after DTT-treatment (P-values range from 0.24 to <0.01). We conclude that the use of DTT to homogenize sputum for dispersion of cells is harmful to cell functions and these cells are hampered for the evaluation of their normal functional characteristics.

Bronchoalveolar Lavage Fluid↗
PubMed2005

Altered expression of immune surface markers in children with recurrent infections of respiratory tract.

Infections of the respiratory tract are very common in young children. They may affect the quality of life and have a great economic impact. On the other hand, respiratory tract infections through MALT system play a positive role in the maturation and development of the immune system. The aim of the study was to examine quantitative and qualitative changes of T and B cells and granulocytes surface molecules in 24 children with recurrent (more than 8 episodes per year) infections of the respiratory tract and in 18 healthy children. The expression of CD2, CD3, CD4, CD8 on T cells; HLA-DR, CD19, CD5/CD20 on B cells, and CD11a/CD18, CD11b/CD18, CD62L, CD16 on granulocytes from peripheral blood was evaluated by a flow cytometry method. We observed a significant increase in the CD5+/CD20+ positive cells on B cells. We also observed a decreased expression of CD11c+/CD18+ cells, CD11a, and CD62L on granulocytes. The expression of other structures on lymphocytes and the CD11b/CD18+ on granulocytes remained unchanged. CD5+/CD20+ cells constitute a filogenetically old population responsible for the production of IgM of low specificity and affinity for specific antigens. The prevalence of this fetal phenotype population may be explained as a delayed maturation of the humoral immune system leading to increased susceptibility to infections. A lower percentage of CD11a may be related to the blockade of that molecule by rhinoviruses.

Antibody Formation↗
PubMed2004

Diagnostic value of different serological tests for tuberculosis in Poland.

The aim of the study was to test the diagnostic accuracy of several serological assays for the diagnosis of tuberculosis (TB) in the Polish population. ELISA based assays detecting: 38 kDa+LAM - MycoM, MycoA and MycoG, 38 kDa - Pathozyme TB complex, 38 kDa+16 kDa - Pathozyme TB complex plus were used. The humoral immune response was analyzed in a group of 319 TB patients (289 adults and 30 children) and in a control group consisting of 66 sarcoidosis cases, 16 cases of mycobacterial infections other than tuberculosis, 35 lung cancer patients, and 70 healthy volunteers. Among the TB patients, there were 267 cases of pulmonary TB and 52 cases of extrapulmonary TB. Sensitivity varied between 32% (IgM) and 63% (IgA) and increased in culture positive tuberculosis and in chronic cases. Specificity was the highest for the tests based on recombinant antibodies (98%). Sensitivity of the IgG test in extrapulmonary TB was comparable with that in pulmonary TB. Overall, sensitivity of the examined tests was lower in children than in adults, but it varied depending on the age and phase of the disease. We conclude that the ELISA-based tests may be a useful tool for improving the diagnosis of TB, especially in adults and in those countries where the prevalence of culture positive and chronic cases is high.

Adolescent↗
PubMed2003

Inhibitory effect of Greenland shark liver oil combined with squalen and arctic birch ashes on angiogenesis and L-1 sarcoma growth in Balb/c mice.

Sharks have been claimed to be resistant to cancer and oil from their livers have been used in Scandinavian folk medicine as anti-tumor drug. Shark liver oil contains 40% or more of squalene. Fish liver oil is also rich in squalene and polyunsaturated n-3 fatty acids. The aim of this work was to determine the anti-angiogenic and anti-tumor effects of these substances, together with another Scandinavian traditional remedy--arctic birch ashes--in Balb/c mice after transplantation of syngeneic L-1 sarcoma. All substances tested, alone or in combinations, significantly diminished cutaneous angiogenesis induced by tumor cells, and tumor growth.

Animals↗
PubMed2002

Angiogenic activity and IL-8 concentrations in peritoneal fluid and sera in endometriosis.

OBJECTIVES: During menstruation endometrial fragments are transported into the peritoneal cavity where they form endometriotic lesions. Angiogenesis is proposed as one of the mechanisms in endometriosis pathogenesis. The aim of the study was to determine the angiogenic activity and interleukin 8 concentrations in peritoneal fluid and sera in endometriosis. METHODS: Angiogenesis was determined in cutaneous assay in Balb/c mice; IL-8 concentrations were measured by ELISA test in sera and peritoneal fluid of 32 control and 56 endometriosis patients. Wilcoxon and Mann-Whitney tests and Spearman rank correlations were used in statistical analysis. RESULTS: Peritoneal fluid and sera from the examined group had higher angiogenic activity and interleukin 8 concentrations. There was correlation found between AFS and neovascularization induced by sera and PF of patients with peritoneal lesions. CONCLUSIONS: Angiogenesis plays an important role in pathogenesis of endometriosis. Although IL-8 takes part in neovascularization, there are other factors modulating angiogenesis in endometriosis.

Animals↗
PubMed2002

Humoral immune response against 38-kDa and 16-kDa mycobacterial antigens in bone and joint tuberculosis.

SETTING: The diagnosis of bone and joint tuberculosis (BTB) is difficult, and diagnostic delays often occur. A reliable serological test detecting anti-mycobacterial antibodies would thus be of some use in this form of the disease. OBJECTIVE: To evaluate the diagnostic accuracy of an assay detecting IgG against 38-kDa and 16-kDa recombinant mycobacterial antigens in BTB. MATERIALS AND METHODS: In a prospective study, serum samples from 124 subjects were examined: 30 BTB cases, 40 non-specific bone and joint infection patients (NSBI), 30 lung cancer patients (LC), and 24 healthy volunteers (HC). An ELISA-based test (Pathozyme TB complex plus) was used. RESULTS: The cut-off level was established at 150 U/ml according to receiver operating characteristic (ROC) curves. The quantified level of sensitivity of the test detecting BTB was 56%, at a specificity of 99%. The positive and negative predictive values were respectively 94% and 88%. Mean IgG level in the BTB group was 470 +/- 761 U/ml (mean +/- SD), and was significantly higher than the antibody level in the control groups (NSBI 58 +/- 42 U/ml, LC 43 +/- 38 U/ml, HC 40 +/- 29 U/ml). CONCLUSION: The test presents an acceptable level of sensitivity and very good specificity in the diagnosis of BTB, and can be used in combination with other methods to increase diagnostic accuracy in this disease.

Adult↗
PubMed2000

[Influence of isoniazid on selected parameters of immunological response].

Isoniazid (INH), antituberculous drug with immunomodulatory properties, have been described as lupus-like syndrome inducer. The development of autoimmunological phenomena results from immunoregulatory disturbances. The goal of this work was to determine the influence of INH on selected parameters of the immune response in vivo and in vitro. In vivo (in B6AF1 mice) the influence of long-term treatment on primary humoral response and cellular response was evaluated. Drug dose was 25 mg/kg. In vitro (using peripheral blood of volunteers) the influence of INH on mitogen induced proliferation, metabolic activity of granulocytes and production of angiogenic cytokines by diverse subpopulation of mononuclear cells was examined. The concentrations tested were 0.5 mg/ml, 5 mg/ml and 50 mg/ml. No effect of INH could be demonstrated on the production anti-SRBC antibodies nor on the cellular response in mice. In vitro INH added to the cell cultures increased PHA and ConA stimulated proliferation. The chemiluminescence of human granulocytes increased in the presence of INH. Drug enhanced production of angiogenic cytokines by human lymphocytes CD4+ and suppressed angiogenic activity of CD8+ cells. The results suggest that INH has strong immunomodulatory properties which may explain its involvement in pathogenesis of lupus-like disease.

Animals↗
PubMed2000

[Value of the immunochromatographic assay for detecting IgG antibodies against 38 kDa mycobacterial antigen in diagnosis of tuberculosis].

Despite of a fast development in the techniques of rapid identification of mycobacteria by molecular genetic techniques, serodiagnosis may be of special values as non-expensive, easy to perform method. Several serodiagnostic tests, principally those using immunoenzymatic (ELISA) methodology are available. The goal of our study was to evaluate one step coloured immunochromatographic assay detecting IgG antibodies against antigen 38 kDa (Rapid Test TB). Our material consisted of 278 serum samples--tuberculosis (n = 155), healthy (n = 36), sarcoidosis (n = 50), lung cancer (n = 25) mycobacterial infections other than tuberculosis (n = 12). Tuberculosis group consisted of new culture positive cases (n = 66), new culture negative cases (n = 23), chronic cases (n = 43) and extrapulmonary TB (n = 23). Specificity of 96% and sensitivity of 54% was obtained. In pulmonary TB sensitivity of 50% and in extrapulmonary TB of 74% was obtained. In chronic cases sensitivity of 70% and in new cases of 40% was received. Sensitivity of 44% in new culture positive cases and 30% in new culture negative cases was obtained. We conclude that immunochromatographic test may be a very useful tool improving tuberculosis diagnosis, especially in extrapulmonary tuberculosis. Strip test may be an interesting alternative as it is an extremely simple, rapid, and cheap technique.

Chromatography↗
PubMed1999

[Usefulness of measuring serum IGG antibodies against A60 mycobacterial antigen for diagnosis of tuberculosis].

Measurement of antimycobacterial antibody may be used as potential diagnostic tool in tuberculosis. The aim of the study was to evaluate the diagnostic value of serum IgG level against A60 mycobacterial antigen measured by ELISA method. Material consisted of 144 persons divided into 5 groups (76 tuberculosis patients, 20 sarcoidosis patients, 17 lung cancer patients, 8 patients with mycobacterial infections other than tuberculosis and 23 healthy controls). In the tuberculosis group there were 50 culture positive cases and 26 culture negative ones, 43 new cases and 32 chronic cases. Positive results were obtained in 51% of tuberculosis patients. Sensitivity increased to 62% in culture positive group and 63% in chronic cases. Specificity of the test was 96%. The results indicate that Immunozyme Mycobacterium test is a valuable tool in tuberculosis diagnosis.

Adult↗
PubMed1998

[Diagnostic value of IgG antibody levels against 38 kDa mycobacterial antigen].

Tuberculosis diagnosis bases on clinical and radiological symptoms and identification of mycobacteria. Accuracy of both methods is limited. Therefore reliable serological test would have considerable advantage. The present study was aimed at evaluating IgG-mediated immune response against specific mycobacterial antigens 38 kDa in group of 200 patients and control subjects. Our material consisted of 104 tuberculosis patients, 25 with sarcoidosis, 24 with lung cancer, 13 with bacterial or fungal pulmonary infection, 8 with mycobacterial infections other than tuberculosis and 26 healthy persons. We used commercially available ELISA based kits (Pathozyme TB-complex). Specificity of 100% and sensitivity of 49% was achieved. Sensitivity increased to 59% in chronic cases and to 52% in culture positive cases. Sensitivity decreased to only 14% in group of new culture negative cases. Measurement of IgG serum level against 38 kDa can be helpful in tuberculosis diagnosis. As the test lacks falsely positive results it indicates its high positive predictive value.

Adult↗