Expression of purinergic P2X7 receptors in subpopulations of peripheral blood mononuclear cells in early-stage of chronic kidney disease.

Chronic kidney disease (CKD) is known as a state of chronic low-grade inflammation, enhancing cardiovascular risk and immunodeficiency. Purinergic signaling has been accepted as a crucial component in the pathogenesis of various diseases, mediating a vast array of biological processes. The P2X7 receptor is one of the important cell surface regulators of several key inflammatory molecules. The aim of the study was to examine the expression of surface P2X7 receptors in subpopulations of peripheral blood mononuclear cells (PBMCs), and to evaluate the promising prognostic markers of inflammation (neutrophil/lymphocyte, Ne/Ly ratio) and cardiovascular risk (monocyte/high density lipoprotein cholesterol, Mo/HDL ratio) in early-stage CKD. The study involved 15 healthy volunteers and 15 non-diabetic patients with CKD stage 2 - 3. PBMCs were isolated from heparinized blood by Ficoll gradient centrifugation. To determine the expression of P2X7 receptors in different subpopulations (CD14+ monocytes, CD3+ T-lymphocytes and CD19+ B-lymphocytes), the cells were stained with FITC-conjugated anti-P2X7. The monocyte, lymphocyte and neutrophil counts were measured in whole blood as a part of routine hemogram. The number of T- and B-lymphocytes was determined by flow cytometry using antibodies anti-CD3-PE and anti-CD19-PE, respectively. The expression of surface P2X7 receptors was 1.4 fold increased in PBMCs of CKD patients compared to healthy volunteers. The expression of P2X7 receptors was 2.1 fold higher in monocytes and 1.5 fold higher in the whole lymphocyte population, with significant increase only in B-cells. The monocyte count, as well as the Ne/Ly and Mo/HDL ratios were also significantly increased. In conclusion, the increased P2X7 receptors expression in monocytes, the monocyte count and the Ne/Ly ratio are manifestations of chronic inflammation already in early stages of CKD. The study also supports recent findings that the Mo/HDL ratio could be used as additional parameter for monitoring cardiovascular risk profile in these patients.

Effects of chronic kidney disease on blood cells membrane properties.

Chronic kidney disease (CKD) is progressive loss of renal function associated among others with increased intracellular calcium concentration. The purpose of this study was to identify the effects of CKD on cell membrane properties such as human red blood cell Ca(2+) ATPase activity, lymphocyte plasma membrane P2X(7) receptor expression and function. This could help us in elucidating the origin of increased calcium concentration in blood cells. We found out Ca(2+) ATPase activity is decreased in early stage CKD patients resulting in altered calcium removal from cytoplasm. By means of flow cytometry we assessed that P2X(7) receptor expression on lymphocyte membrane is 1.5 fold increased for CKD patients. Moreover, we detected an increased uptake of ethidium bromide through this receptor in CKD at basal conditions. It means CKD lymphocyte membranes contain more receptors which are more permeable thus allowing increased calcium influx from extracellular milieu. Finally, we can state alterations in blood cell membranes are closely linked to CKD and may be responsible for intracellular calcium accumulation.

The kinetics of cell surface receptor expression in children perinatally exposed to polychlorinated biphenyls.

Exposure to polychlorinated biphenyls (PCBs) during pre-natal and early life can alter normal immune system development. Blood specimens from newborns, 6-, and 16-month-old infants were collected in the Michalovce and Svidnik/Stropkov districts, areas with, respectively, high and low environmental PCB contamination, and lymphocyte receptor expression was evaluated by multi-color flow cytometry. The results indicate that the percentage of lymphoid dendritic cells (DC) and naïve/resting T-lymphocytes were significantly increased at 6-months in Michalovce as compared to the same cell types in cord blood samples (p < 0.001), whereas natural regulatory T-lymphocytes and suppressor inducer T-lymphocytes were reduced (p < 0.001). Overall, a positive linear correlation of terminally differentiated effector memory (TEM) T-lymphocyte population with age, but a negative linear correlation for myeloid DC from birth to 6-months in both regions were found. Michalovce samples indicated significantly higher expression of memory T-lymphocytes (birth, 6(th), and 16(th) month), TEM T-lymphocytes (birth and 6(th) month), and lymphoid DC (6(th) month) compared to the Svidnik/Stropkov regions. After adjustment for relevant covariates, such as maternal age, parity, season of birth, breastfeeding, birth weight, and gender, the myeloid DC, suppressor inducer T-lymphocytes, truly naïve helper/inducer T-lymphocytes, and TEM T-lymphocytes remained significantly different between districts in cord blood samples. The multivariate analysis models for 6- and 16-month samples showed district differences in all cellular determinants, except for lymphoid DC and macrophage-like cells. This study provides the first evidence that pre-natal and early post-natal exposure to PCBs affects the dynamics of cell surface receptor expression on lymphoid DC and DC-like cells, suggesting impaired immunologic development following pre-natal and early post-natal PCB exposure.

Dynamics of lymphocyte subsets in children living in an area polluted by polychlorinated biphenyls.

Immune system development, particularly in the pre-natal and early post-natal periods, has far-reaching health consequences during childhood, as well as throughout life. Exposure to poly-chlorinated biphenyls (PCBs) during pre-natal and early life has been previously associated with changes in the incidence of infectious and allergic diseases in children, and humoral immunity alterations. Lymphocyte immunophenotyping is an important tool in the diagnosis of immunologic and hematologic disorders. This study used a lysed whole blood method for analysis of lymphocyte sub-populations in samples from children born and living in two districts: a highly-contaminated area (Michalovce) and one (Svidnik/Stropkov) with ≈ 2-fold lower environmental PCB levels. The percentages of B-lymphocytes (CD19(+)), activated HLADR(+)CD19(+) cells, and CD8(+) T-lymphocytes significantly increased at 6- and 16-months-of-age in both selected regions as compared to in cord blood values (p < 0.001). Levels of CD3(+) cells increased significantly (from 61 to 65%) in samples from Michalovce (p < 0.01). Levels of CD4(+) T-lymphocytes declined 10% among 16-month-olds in both regions (Michalovce at p < 0.001 and Svidnik/Stropkov at p < 0.01). Natural killer (NK) cell levels decreased 50% in Michalovce 6- and 16-month-old children and 42% among 6-month-olds in Svidnik/Stropkov (p < 0.001). Compared with the less-contaminated region, Michalovce samples showed significantly higher expression of CD3(+) T-lymphocytes, B-lymphocytes, and activated B-lymphocytes, whereas NK cells were less expressed. Even after adjustment for selected covariates, e.g., maternal cigarette smoking, age, parity, ethnicity, birth weight, and gender of infant, the levels of CD19(+), HLADR(+)CD19(+), and CD3(-)CD(16 + 56)(+) cells were seen to remain significantly different between the districts. These results showed that early-life environmental PCB exposure was associated with fluctuations in major lymphocyte subsets in children, suggesting that there is a post-natal immune system response to PCB exposures.

Dose reduction using bismuth shielding during paediatric CT examinations in Slovakia.

Considering the massive increase of computer tomography (CT) examinations in Slovakia during the last 10 y, it can be expected that a higher radiation load may be observed in the Slovak population. Since child population is more sensitive to radiation than adult population, a monitoring has started to see how high the radiation dose is for paediatric patients during CT examinations in chosen departments in Slovakia. The CT examination of the head is one of the most frequently done examinations in Slovakian departments and that is why measurements were done to clarify how usage of bismuth shields for eyes and thyroid can affect the eye and thyroid doses. For simulation, 215 thermoluminescent dosimeters were exposed on anthropomorphic phantom of a child with and without usage of bismuth shields. The result was that only two of the three chosen departments confirmed a reduction. On the other hand, one of the departments confirmed that the reduction can be up to 56-65 %, which is significant.

The relationship between cell surface markers, cytokines, ageing, and cigarette smoking.

The purpose of this study was to investigate the modulation of selected cell surface markers and proinflammatory cytokines production in relation to ageing, and cigarette smoking. The analysis of cell surface receptors was performed by the flow cytometry and cytokines levels were evaluated by the sandwich enzyme immunoassays. We found a decreased expression of CD69, CD28, CD11b, CD95 markers in old population compared to young people (p<0.05; p<0.001). The memory CD45RO lymphocytes were markedly expanded in older population in comparison to young donors (12.93+/-5.92 %, p<0.001) and the selectin CD62L was significantly increased on granulocytes in aged people (p<0.05). Our findings demonstrated an augmented level of CD3 and CD28 on lymphocytes in smokers (p<0.05; p<0.005). The significant depression of CD16+56 molecule was recorded in smokers (10.86+/-0.80%) when compared to non-smokers (14.44+/-0.46; p<0.05). Our results showed a significantly diminished levels of interleukin (IL)-1beta (1.93+/-0.48 pg/ml), and increased levels of IL-6 and tumor necrosis factor (TNF)-alpha in elderly population compared to young people (p<0.05; p<0.001). The present data support previous suggestions that senescence and cigarette smoking may contribute to changes in the immune system activity, resulting in altered cell surface marker expression and cytokine levels (Tab. 1, Fig. 3, Ref. 81). Full Text (Free, PDF) www.bmj.sk.

Simultaneous flow cytometric evaluation of phagocytosis and oxidative burst in human polymorphonuclear cells.

Phagocytosis and oxidative burst (OXIBURST) activity of human polymorphonuclear cells (PMNs) has been simultaneously measured directly in whole blood samples. The ingestion of yeast was assessed by the phagocytosis activity (FA) and phagocytosis index (FI), and the respiratory burst of PMNs was determined as dihydroethidine (DHE) oxidation. We received comparable results in the ingestion of yeast cells by PMNs using either light microscopy (77.31+/-7.56) or flow cytometry detection method (78.26+/-5.14). The significant differences (p<0.05) in FI and OXIBURST activity were find in the patients (2.29+/-0.29 and 14.67+/-3.99, respectively) when compared to healthy donors (1.64+/-0.21 and 32.38+/-14.94, respectively). The two-color flow cytometric procedure permits measurement of two different functions of neutrophils in one step. This flow cytometric procedure is simple, rapid and has the potential to be an alternative assay to test leukocyte function. (Fig. 3, Ref: 30.)

Implementation of QA and QC standards in radiology in Slovakia.

The Slovak government organizes the radiation protection policy through its regulatory authorities within the Ministry of Health, on a central level. The health protection regulations are compatible with international standards and recommendations of the ICRP and the EC. The general requirements on quality assurance (QA) and quality control (QC) (acceptance, constancy and routine tests), the guidance levels for various types of radiological examinations and the instructions for optimisation procedures are supported by Slovak technical standards, compatible with European standards. But the QA/QC process, as well as the training of the staff, needs improvement. The Slovak Medical University participates in the QA implementation through organising and managing national audits for control of the QA programme. In this paper, we present results of patient dose measurement studies carried out in the Slovak Republic, in the framework of activities of the Slovak Commission of QA established by the Slovak Ministry of Health.

Autofluorescence (safe) bronchoscopy and p21/ki-67 immunostaining related to carcinogenesis.

Archival biopsy materials from 20 randomly selected asymptomatic volunteers from the Czech uranium miners (CZ UM) risk group (n=98) were examined for p21 and ki-67 immunostatning. There were 16 areas with normal respiratory epithelium and 22 areas with bronchial intra-epithelial neoplasia (IEN). Normal and IEN areas were identified by autofluorescence (System Autofluorescence Endoscopy, SAFE-1000) and monitored during 1998-2002. The majority of specimens from areas with normal autofluorescence intensity with ciliated columnar bronchial epithelium showed strong predominantly cytoplasmic p21 positivity. The SAFE monitoring divided areas of decreased autofluorescence intensity with early stage IEN lesions into two groups. Persistent lesions (P)-showing a spectrum of p21 cytoplasmic staining ranging from negative or isolated negativity to weak or moderate positivity combined with higher proliferative capacity proved by ki-67 nuclear staining. Disappearing lesions (D)-showing strong cytoplasmic p21 positivity and negative ki-67 staining. The IEN lesions were classified into three groups based on p21/ki-67 immunostaining: proliferative lesions at risk (R) with low or without p21 plasma immunostaining combined with high ki-67 nuclear reactivity; ambiguous lesions (A) including cases combining strong p21 cytoplasmic positivity with high ki-67 nuclear reactivity or p21 cytoplasmic negativity with ki-67 negativity staining patterns; the quiescent lesion group (Q) was characterized by strong p21 cytoplasmic positivity and negative ki-67 immunostaining.

Human eosinophils as potent inflammatory cells and their apoptosis.

A significant association was established between the eosinophil and a number of disease conditions, including helminthiasis, allergy, asthma, drug hypersensitivity, certain neoplasias, and graft rejection. Activation of eosinophils and release of proinflammatory lipid mediators, cytokines, free oxygen radicals, highly-charged cationic proteins contribute to the onset and maintenance of tissue inflammation. Eosinophil accumulation in blood and tissues has been related to a defect in their apoptotic death. Decisive events during the apoptotic process involve mitochondrial permeabilization and caspase activation. Clearance of apoptotic cells depends on the ability of phagocytes to recognize their cell targets and, subsequently, to engulf them. (Tab. 2, Ref. 32.)

Antiasthmatic effects of nedocromil sodium.

Accumulating data indicate that bronchial asthma is a chronic inflammatory disease. Airway inflammation and it's control became a principal focus in asthma treatment. Nedocromil sodium is chemically nonsteroidal anti-inflammatory agent for the treatment of mild to moderate asthma. The aim of the study was to determine the effects of NS on bronchial hyperresponsivness and eosinophil activation markers isolated from peripheral blood of asthmatics with mild intermittent asthma. Twenty nine patients of both sexes (17 women, 12 men) with average age of 34 years were recruited into the clinical open study. Bronchial responsivness was assessed by metacholine challenge test prior to starting therapy with NS (preparation Tilade mint aer) and 3rd week and 9th week of follow up. Baseline lung function tests were performed at intervals before treatment and at 3rd and 9th week, respectively. Eosinophil activation markers were determined before and after 3rd and 6th week. Assessement was done by flow cytometry using standard monoclonal antibodies. Bronchial responsivness decreased significantly at 3rd and 9th week of follow up (provocation dose--PD20 increased significantly, p < 0.05, p < 0.02, respectively). Improvements of baseline lung function tests were observed in majority of parameters: FVC (p < 0.01), FEV1 (p < 0.01), FEV1/FVC (p < 0.01), MEF 25 (p < 0.03), MEF 50 (p < 0.01), MEF 25-75 (p < 0.01), PEF (p < 0.01) after 3rd week, however the enhancement of improvement was seen in majority of parameters after 9th week of the study. Significant reduction of eosinophil activation markers expression was noticed: CD69 (p < 0.05, p < 0.01) and HLA DR (p < 0.05, p < 0.05) after 3rd and 6th week, respectively and CD66 (p < 0.05) after 3rd week and CD81 (p < 0.05) after 6th week of follow up. NS possessed complex antiasthmatic effects resulting in decrease of bronchial responsivness and reduction of eosinophil activation markers in mild asthmatics. The tolerance of the drug was good and no adverse effects have been reported. NS is effective prophylactic drug recommended for use in both adults and children in long-term management of mild asthma. (Tab. 2, Fig. 1, Ref. 27).

Immunophenotyping of leukocytes in peripheral blood of BALB/c mice infected with mouse herpesvirus isolate 72.

We characterized leukocytes in peripheral blood of BALB/c mice infected with mouse herpesvirus isolate 72 (MHV-72) representing an isolate of mouse herpesvirus strain 68 (MHV-68, species Murid herpesvirus 4, genus Rhadinovirus, subfamily Gammaherpesvirinae, family Herpesviridae) (van Regenmortel et al., 2000). In acute infection (up to day 30 post infection (p.i.)) the number of CD8+ T cells increased, reaching a maximum at day 11 p.i. This increase correlated with that of CD4+ T, activated CD 19+ B and natural killer (NK) cells. At day 30 p.i. the numbers of CD4+, CD8+, CD14+ and CD19+ cells decreased to normal values. A similar increase in the number of these cells was observed at day 730 p. i. In the course of persistent infection (after day 30 p.i.) some of the mice developed a leukemia-like syndrome characterized by an increase in the number of leukocytes and appearance of atypical, blastic immature forms of leukocytes. The latter forms of leukocytes were characteristic by an increased amount of argyrophilic proteins. These results show further similarities between MHV-72 (another isolate of MHV-68) and EBV infections and justify the use of MHV-68 or MHV-72 as an appropriate mouse model for the study of EBV infection of humans.

Effects of selenium supplementation on expression of adhesion molecules in corticoid-dependent asthmatics.

Selenium (Se) deficiency attenuates the host immune response, thereby increasing the risk of bacterial and viral infections. We have examined the effects of selenium supplementation (SeS) in corticoid-dependent asthmatics (CDAs) with lowered circulatory Se status. Twenty CDAs (10 males and 10 females, average age 54.5 yrs) were enrolled into the study. The average duration of the disease was 10 yrs. The asthmatics were receiving 200 micrograms of Se per day for a period of 6 months, in addition to regular treatment with inhaled corticosteroids and beta-agonists. The expression of adhesion molecules (CD11a, CD11b, CD18, CD49d, CD54, CD62L) on peripheral blood mononuclear cells (PBMCs) of asthmatics and the expression of E- and P-selectins, ICAM-1, VCAM-1 on cultured human umbilical vein endothelial cells (HUVEC) after stimulation with PBMCs from CDAs before and after 3 and 6 months of SeS were assessed by standard monoclonal antibodies and analyzed by flow cytometry. The concentrations of soluble adhesion molecules P-selectin, E-selectin, ICAM-1 and VCAM-1 were determined by ELISA method. The expression of adhesion molecules on PBMCs: After 3- and 6-months of SeS, a decreased expression of molecules CD11a, CD11b and CD62L was observed (p < 0.02, p < 0.005, p < 0.003). No changes were seen in the expression of CD18, CD49d except for the increased expression of CD54 (p < 0.005). Modulation of adhesion molecules expression on HUVEC: We observed a significant increase in VCAM-1, P- and E-selectins expressions in the group of asthmatics without SeS in comparison with the control group (p < 0.05, p < 0.01, p < 0.05). During SeS a significant decrease in molecules VCAM-1, E-selectin (after 3 months) (p < 0.05, p < 0.05) and P-selectins and ICAM-1 (after 6 months) (p < 0.05, p < 0.01) were observed. Soluble adhesion molecules: After 3 months of SeS we noticed a significant decrease in VCAM-1 and P-selectin expressions (p < 0.05, p < 0.05) and after 6 months the level of VCAM-1 decreased (p < 0.01). The effect of Se on the adhesion molecules expression in endothelial cells in vitro experiments: Se blocks the expression of adhesion molecules stimulated by IFN-gamma in a dose-dependent way after addition of Se into a culture of endothelial cells. Concentration of 10 micrograms/ml inhibits the increase in expression of ICAM-1 (p < 0.05) but not that of VCAM-1, E- or P-selectins. The inhibition of expression in Se concentration of 10 micrograms/ml is over 80% (p < 0.01). Our data demonstrate that Se is able to affect the adhesion molecules expressions that are crucial in the inflammatory process. (Fig. 5, Ref. 22.)

The influence of selenium supplementation on the immunity of corticoid-dependent asthmatics.

UNLABELLED: Selenium (Se) is a trace element that is essential for immune functions, and protects the immune system from oxidative damage. AIM: The aim of the pilot clinical study was to assess the influence of selenium supplementation (SeS) on the selected immune parameters analyzed from peripheral blood of corticoid-dependent asthmatics (CDAs). MATERIAL AND METHODS: Seventeen CDAs aged from 30 to 74 years (7 females, 10 males) with suboptimal levels of Se in plasma were enrolled into the study. The follow up of SeS lasted 96 weeks. It is daily dose was 200 micrograms (2 x 2 tbl daily, 1 tbl contained 50 micrograms of Se). Before (-4 weeks) and after the 12th, 48th, 72nd and 96th weeks of SeS, the following parameters were observed: Epitopes EG1, EG2 expressed on intracellular eosinophil (Eo) cationic protein and eosinophil peroxidase, the numbers of CD3, CD4, CD8, CD19 and CD3 HLADR positive T lymphocytes (Ly), lymphocyte blastogenesis test (LTT) with mitogens concanavalin A, (Conc A) phytohemaglutin (PHA), the levels of C3, C4 complement components, activation of complement by classic and alternative pathways (CP50, AP50), the levels of immunoglobulins (Ig) G, A, M and total IgE, circulating immune complexes (CIC). RESULTS: Epitopes EG1 and EG2 in cytoplasma of Eo decreased significantly after 12 weeks of SeS, (p < 0.01) and 96 weeks of follow up. In parameters of T cell mediated immunity the relative number of CD3 HLADR+ T Ly increased after 24, 48 and 96 weeks of SeS (p < 0.0008, p < 0.009, p < 0.07). Proliferative activity of T Ly to mitogenes PHA and ConcA in LTT decreased significantly after 12, 48, 72 and 96 weeks of SeS (p < 0.0005, p < 0.009, p < 0.04, p < 0.02, respectively). In humoral parameters activation of CP50 decreased after 24, 72 and 96 weeks of SeS to the reference range (p < 0.001, p < 0.03, p < 0.02) and AP50 after 96 weeks, respectively (p < 0.02). The levels of IgG elevated after 24 weeks (p < 0.02), IgA after 24, 48 weeks (p < 0.0007, p < 0.02, respectively). The level of total IgE significantly decreased after 96 weeks of SeS (p < 0.003). CONCLUSION: Our pilot clinical study with the CDAs demonstrates the significant changes particularly in functional parameters of both cellular and humoral types of immunity. These results support the immunomodulating effects of SeS. (Tab. 5, Ref. 15.)

Levels of coenzyme Q10 in asthmatics.

BACKGROUND: The contribution of free oxygen radicals in the pathogenesis of bronchial asthma is generally accepted. The modulation of antioxidative defence by supplementation with antioxidants represents additive approach in complex management of the disease. The aim of the study was to assess the levels of coenzyme Q10, alpha-tocopherol, beta-carotene and malondialdehyde (end-stage parameter of lipid peroxidation) in asthmatics (As). METHODS: Fifty six As (15 males and 41 females) aged from 19 to 72 yrs (mean age 46 yrs) were enrolled into the study. The control group comprised of 25 healthy volunteers (16 males, 9 females) aged 25-50 years. RESULTS: Concentrations of CoQ10 and alpha-tocopherol, decresed significantly both in plasma and whole blood, compared with healthy volunteers (p < 0.009, p < 0.004; p < 0.035, p < 0.001, respectively). The level of MDA was elevated, but not statisticaly significantly. No changes were seen in beta-carotene levels. Positive correlation was found between concentrations of CoQ10 and alpha-tocopherol. CONCLUSION: Our results suggest possible contribution of suboptimal concentrations of CoQ10 on antioxidative dysbalance in As and provide rationale for its supplementation with clinical evaluation. (Tab. 2, Fig. 1, Ref. 39.).

Detection of antiendothelial cell antibodies in patients with connective tissue diseases by flow cytometry and their relation to endothelial cell activation.

Antiendothelial cell antibodies (AECA) have been detected by flow cytometry analysis in 23 out of 80 patients with connective tissue diseases. Ten out of 19 serum samples from patients with systemic lupus erythematosus (SLE) were positive. These antibodies were not detectable in healthy donors. We examined the capacity of serum samples to induce endothelial cell activation by modulating cell adhesion molecule expression on human umbilical vein endothelial cells. We found that sera from both AECA-positive and AECA-negative patient groups induced a significantly higher expression of E-selectin compared to healthy controls (P<0.05). There were no differences in the ICAM-1 on VCAM-1 expression. Our data suggest that increased E-selectin expression in activated endothelium in patients with various connective tissue disorders is not related to the production of AECA.

Inflammatory mechanisms involving neutrophils in chronic venous insufficiency of lower limbs.

BACKGROUND: It is supposed that an inflammatory reaction is one of the major factors responsible for the chronic venous insufficiency (CVI) of lower limbs which cause leg ulcers. OBJECTIVES: The main objective of the present study was to determine the differences in the levels of typical inflammatory mediators and markers produced by neutrophils of patients with CVI and normal control subjects. SUBJECTS AND METHODS: 26 patients with CVI and 39 clinically healthy subjects were included in the study. In peripheral neutrophils of both groups the production of superoxide, total reactive oxygen intermediates and activities of lysosomal enzymes were measured together with the expression of 8 adhesion molecules. RESULTS: Increased formation of superoxide by patient neutrophils and activities of elastase in both neutrophils and serum of patients were demonstrated. On the contrary, activities of myeloperoxidase and beta-D-glucuronidase were decreased in patient neutrophils. Comparing to control group adhesion molecules CD11b, CD18, CD31, CD49d, CD54 and CD62L were increased on the surface of patient neutrophils whereas no differences were observed in the expression of CD11a abd CD15. CONCLUSION: The neutrophils of patients with CVI are primed and/or activated because they are able to release higher amount of superoxide, lysosomal enzymes and express elevated number of adhesion molecules. It may serve as one of the important evidences of an inflammatory mechanism involved in the pathogenesis of chronic venous insufficiency. (Tab. 3, Ref. 27.)

Detection of intracellular cytokines during antioxidant supplementation in corticoid-dependent asthmatics and modulation of adhesion molecule expression on cultured endothelial cells.

In this study, we report on the interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) cytokine responses to phorbol myristate acetate (PMA) + ionomycin-stimulated CD3+ lymphocytes in asthmatic subjects when compared with normal donors. There was a significantly lower production of intracellular IFN-gamma in asthmatic patients. No difference was found for IL-4 production between these two groups. After administration of a multivitamin-mineral supplement containing selenium, zinc, vitamin A, vitamin B6, vitamin C, and vitamin E for 6 mo, a significant increase in the percentage of CD3+/IL-4 positive cells (p < 0.05) was found. The induction of endothelial cell adhesion molecule (CAM) expression in cultured human umbilical vein endothelial cells (HUVEC) and whole-blood mixture was studied using flow cytometry. The ICAM-1 and VCAM-1 expressions were higher in the patients than in control donors (p < 0.05). There is a correlation between the increased percentage of CD3+/IFN-gamma positive cells and reduced endothelial ICAM-1 and VCAM-1 expression after 6 mo of intervention period. No apparent effect of supplementation on CAM expression was found, suggesting that these changes do not arise from an antioxidant mechanism. This newly developed whole-blood technique for the assessment of CAM expression can be of use for monitoring therapy in inflammatory diseases.

[The vascular endothelium in inflammatory processes in vivo and in vitro]. / Cievny endotel v zápalovom procese in vivo a in vitro.

Endothelial injury occurs in atherosclerosis, infectious, rheumatic and vasculitic processes, leading to activation of transcription factors and endothelial expression of various cytokines and adhesion molecules. Endothelial cell cultures represent a valuable tool in research activities, with emphasis on the principal characteristics of angiogenesis, inflammatory response, transduction signals and endothelial functionality. In the laboratory of tissue cultures we prepared primary endothelial cultures by their isolation from the umbilical vein. This model system has been used to investigate the endothelial activation in vitro, adhesion alterations of immunocompetent cells to endothelium, adhesion molecule expression in the disease course monitoring and anti-inflammatory treatment. (Tab. 1, Fig. 5, Ref. 45.)

[The role of adhesion molecules in the immune system]. / Uloha adhezívnych molekúl v imunitnom systéme.

Adhesion molecules play a major role in many biological functions. They are crucial in the development of embryo into formed organism; later they mediate many physiological and pathological functions. From the immunological point of view they are involved in virtually every process of cell interactions, involving thymic selection and antigen priming, antigen recognition and cell activation, cytotoxicity and lymphocyte recirculation. This review focuses mainly on the role of adhesion molecules in close contact between the cells, crucial for the inflammatory and immune responses. (Tab. 3, Fig. 1, Ref. 29.)