Low serum vitamin D levels are associated with a low percentage of TREM-2+ monocytes in low-grade gliomas and poorer overall survival in patients with high-grade gliomas.

INTRODUCTION: Anti-inflammatory effect of vitamin D (VD) could be beneficial in improving the survival of glioma patients. The aim of our study was to analyse the serum levels of vitamin D in glioma patients and to find an association with the prognosis of glioma patients and other investigated parameters. MATERIAL AND METHODS: The study included 63 patients with gliomas. Percentage of CD14+ monocytes, TREM-1+ and TREM-2+ monocytes were determined by flow cytometry, serum levels of 25(OH)D were evaluated by electrochemiluminescent binding test. RESULTS: Six patients out of 63 had normal levels of VD. A significant difference in the overall survival (OS) in the patients with severe VD deficiency, VD deficiency and insufficiency in grade IV was found. In grade II and III, the levels of vitamin D positively correlated with the percentage of TREM-2+ monocytes, and in grade II also a negative correlation of VD with TREM-1/TREM-2 ratio was observed. CONCLUSION: Levels of VD could influence the prognosis of patients with high-grade gliomas. Serum level of 25(OH)D in low-grade gliomas positively correlated with the percentage of anti-inflammatory acting TREM-2+ monocytes and negatively with TREM-1/TREM-2 ratio. This could be protective against the progression to high-grade glioma, because TREM-2 is associated with protective functions such as: tissue repair, control of local inflammation, or phagocytosis (Tab. 4, Fig. 4, Ref. 79).

Biofilm-producing potential of urinary pathogens isolated from chronic and recurrent urinary tract infections and impact of biofilm on gentamicin and colistin in vitro efficacy.

AIM: The presented study was to compare in vitro biofilm production by bacterial strains from chronic/recurrent and from acute non-complicated UTIs. The activity of gentamicin and colistin on biofilm form of these strains has also been detected, with goal to predict the gentamicin and colistin therapeutic efficacy in the antimicrobial treatment of patients with a suspected presence of biofilm in urinary tract. MATERIAL AND METHODS: The group of 40 bacterial strains repeatedly isolated from patients with chronic or recurrent UTIs was compared with the group of 40 strains from acute UTIs. Both groups contained comparable number of strains of Escherichia coli, Klebsiella spp., Proteus mirabilis and Pseudomonas aeruginosa. Biofilm production was assessed by method in polystyrene microtiter plate. The MIC and MBC values of gentamicin and colistin were detected by broth microdilution assay. The minimal biofilm inhibitory (MBIC) and biofilm eradication concentrations (MBEC) were tested by microdilution method. Non-inactivated biofilm-associated bacteria were detected after overnight incubation in broth medium free of antimicrobials. The statistical analysis of results was performed by Fisher's exact test and by Student's t-test. RESULTS: Biofilm was produced by 90% strains from chronic UTIs, but only by 52% of strains from acute UTIs (p = 0.0004). In the biofilm producing strains, the MBIC values of gentamicin reached from four to 256 mg/L, the MBIC levels of colistin from two to 64 mg/L. The minimal biofilm eradicating concentrations were even higher: for gentamicin from eight to > 512 mg/L, and for colistin from 32 to > 512 mg/L. The differences between MIC and MBIC/MBEC levels were statistically highly significant (p < 0.0001). Presumably, the therapeutic success of parenterally applied gentamicin or colistin on biofilm-related urinary tract infections would be, without respect to the high concentration of gentamicin or colistin achievable in urine during parenteral application, rather unpredictable. Local intravesical instillation would allow for achieving higher gentamicin and colistin concentrations; however, there is need for interpretation criteria for MBEC values concerning therapy, as well as for clinical studies allowing for application of those values to predict clinical success of therapy. CONCLUSIONS: Laboratory detection of biofilm production and evaluation of the MBIC/MBEC values of antimicrobials for strains producing biofilm might be a valuable complement to the microbiologic diagnostics of chronic and recurrent UTIs. It might provide valuable information for more reliable individualised therapy and so decrease the risk of emergence and selection of multiresistant strains during repeated and non-eradicating therapy of chronic and recurrent UTIs.

Decreased plasma levels of 25(OH)D in multiple sclerosis patients. Correlation with disease severity expressed by EDSS, MSSS, progression index and Herbert´s scale severity grade.

OBJECTIVES: Multiple sclerosis is a chronic inflammatory and autoimmune demyelinating disease of the brain and spinal cord. Vitamin D has anti-inflammatory and anti-Th1, Th17 activities, activates the function of regulatory T cells, shifts the immune response towards Th2, so it might be favorable for downregulation of the disease pathogenesis, and if inflammation and Th1 and Th17 immunity are hyperactivated. The aim of our study was to highlight the role of vitamin D in multiple sclerosis pathogenesis. METHODS: We investigated 178 patients with multiple sclerosis. Plasma levels of 25(OH)D and HMGB1 were investigated. RESULTS: Despite a regular use of VD by patients, the plasma levels of 25(0H)D were significantly decreased in 57% of them, 14.1% had VD deficiency (level of 25(OH)D < 20 ng/mL) and more than 6 % of patients had VD severe deficiency with the plasma level of 25(OH)D < 12 ng/mL. The level of 25(OH)D negatively correlated with the severity of the disease (EDSS, index of progression, duration of the disease) and negative association was found also with Herbert´s six severity grades. HMGB1 levels were higher in patients (p < 0.0001). CONCLUSION: Our result showed that vitamin D deficiency plays a role in multiple sclerosis pathogenesis. We believe that administration of vitamin D to patients at a sufficient dose providing a physiological level of vitamin D could have a positive effect on the course of the disease. However, regular monitoring of vitamin D levels is required, which should be at least within 30-75 ng/mL, and even more, but below the toxicity limit (Tab. 6, Ref. 66).

Expression of nuclear retinoic acid receptor in peripheral blood mononuclear cells (PBMC) of healthy subjects.

In vertebrates, both nuclear all-trans and 9-cis retinoic acid receptors (RAR and RXR) belonging to the steroid/thyroid/retinoid nuclear receptor superfamily play a crucial role in the vitamin A action. Qualitative analysis of all known RAR or RXR subtypes in both pooled and non-pooled peripheral blood mononuclear cells (PBMC) from healthy human subjects has been performed by reverse transcription and polymerase chain reaction (RT-PCR). Our data, based on qualitative RT-PCR analysis has shown that human PBMC are capable to express RAR alpha, RAR gamma, RXR alpha, and RXR beta.

Effect of hyperthermic water bath on parameters of cellular immunity.

Effects of hyperthermic water bath on selected immune parameters (lymphocyte subpopulations, natural killer (NK) cell counts and their activity) were studied in a group of 10 volunteers. Application of hyperthermic water bath (both topical and whole-body) was followed by a significant reduction of relative B lymphocyte counts. Whole-body hyperthermic water bath reduced relative total T lymphocyte counts, increased relative CD8+ T lymphocyte and NK cell counts and increased NK activity. Whole-body hyperthermic bath increased somatotropic hormone (STH) activity in eight out of 10 volunteers; higher relative counts of CD8+ lymphocytes and NK cells were observed compared with the group of volunteers not responding to hyperthermic water bath by STH secretion. In five volunteers STH was released in response to local hyperthermic water bath and the NK activity of lymphocytes also increased but their relative counts did not. The results suggest that these increases in CD8+ lymphocyte and NK cell counts are probably dependent on increased STH production.

Plasma cytokine concentration and the cytokine producing ability of whole blood cell cultures from healthy females with pharmacologically induced hyperprolactinemia.

We investigated the in vitro effect of domperidone-induced hyperprolactinemia on plasma cytokine concentration and blood leukocyte cytokine production in healthy female volunteers. No changes were found in the plasma concentration of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, IL-10, IL-6 and IL-13 during hyperprolactinemia when compared with control values. Using unseparated blood leukocytes, we found that the spontaneous production of IL-6 (4-8 h) and transforming growth factor (TGF)-beta 1 (2-4 h) was significantly decreased and that the in vitro stimulated production of IFN-gamma (2-8 h) and TNF (4 h) was significantly increased compared with control. Our data concerning the increased IFN-gamma and TNF producing capacity of unseparated leukocytes during pharmacologically induced hyperprolactinemia strongly support the possibility that the lymphocyte production of these cytokines can be rapidly amplified by prolactin via a priming mechanism.

[Changes in the capacity of T-lymphocytes for spontaneous recovery of selected differentiation antigens in relation to age]. / Zmena schopnosti spontánnej znovuobnovy vybraných diferenciacných antigénov T-lymfocytov v závislosti od veku.

BACKGROUND: During physiological ageing changes of the immune system take place at several levels. The objective of the submitted work was to compare the ability of spontaneous restoration of selected differentiation antigens on lymphocytes in the peripheral blood stream after previous trypsin treatment in a group of healthy elderly and adult subjects. METHODS AND RESULTS: Twenty-four adults were examined (19-59 years) and 36 elderly subjects (60-90 years). Isolated lymphocytes from the peripheral blood stream were treated with trypsin and then incubated in a cultivation medium. The authors investigated the capacity of restoration of differentiation antigens CD2, CD4, CD8 and CD45RA. Antigen CD2 was not restored in any of the investigated groups to original levels. However the difference between its expression on lymphocytes before trypsin treatment and on lymphocytes after 16-hour incubation was higher in the elderly subjects 16% (p < 0.001) than in the group of adults 7% (p < 0.01). Restoration of antigen CD4 was in both investigated groups almost equal. The number of CD8+ T-lymphocytes was in elderly people lower (p < 0.05), spontaneous restoration of antigen CD8 did not differ among the investigated groups and reached in both instances the baseline value. Antigen CD45RA was restored more slowly in elderly subjects, the difference between groups was at borderline of statistical significance (p < 0.0595). CONCLUSION: From the results ensues that during physiological ageing the ability of spontaneous restoration of antigens CD2 and CD45RA declines but not of antigens CD4 and CD8. So far there is no unequivocal explanation why this change occurs, it is probably conditioned by several factors. Investigation of these changes and an attempt to influence them can help to understand age-conditioned immunological dysregulation, its consequences and the possibility to influence them by treatment.

The hypothalamic-pituitary response in SLE. Regulation of prolactin, growth hormone and cortisol release.

It has been suggested that neuroendocrine regulation plays an important role in the pathogenesis and activation of autoimmune diseases. The aim of this investigation was to clarify the hypothalamic-pituitary response to a well-defined stimulus under standardised conditions in patients with SLE. Plasma concentrations of prolactin (PRL), growth hormone (GH) and cortisol were determined in venous blood drawn through an indwelling cannula during insulin-induced hypoglycaemia (0.1 U/kg b.w., i.v.) in ten patients and in 12 age-, gender- and weight-matched healthy subjects. Basal PRL concentrations were higher in patients vs healthy controls (12 vs 6 ng/ml, P < 0.01), though still within the physiological range. Insulin-induced plasma PRL and GH were significantly increased both in patients and healthy subjects; however, the increments or areas under the curves were not different in the two groups. Plasma cortisol response showed moderate attenuation in patients. Sensitivity of pituitary lactotrothrops to thyrotropin-releasing hormone (TRH) administration (200 microg, i.v.) was the same in patients and control subjects. In SLE patients with low activity of the disease the sensitivity of pituitary PRL release to TRH administration remained unchanged. The hypothalamic response to stress stimulus (hypoglycaemia) was comparable in patients and healthy subjects.

Relationship between endocrine, immune, and clinical variables in patients with systemic lupus erythematosus.

OBJECTIVE: To assess the frequency of increased plasma prolactin (PRL) in patients with systemic lupus erythematosus (SLE) and to evaluate its relationship to other hormonal and immune variables. METHODS: Thirty-five patients with SLE with various levels of disease activity were studied. Plasma PRL, cortisol, growth hormone (GH) were determined by radioimmunoassay and interleukin 6 (IL-6) by ELISA: SLE activity was evaluated using the European Consensus Lupus Activity Measurement (ECLAM). RESULTS: Increased plasma PRL concentration (> 20 ng/ml) was recorded in 11 patients (31%). No correlation was found between plasma PRL and GH, IL-6, cortisol, or C-reactive protein, nor was any significant correlation observed between plasma PRL and the ECLAM score. Patients with hyperprolactinemia were, however, found to have been treated with higher doses of prednisone therapy than patients with normal plasma PRL. Further analysis of the relationship of plasma PRL and therapy showed that patients with SLE selected by the attending physician for prednisone therapy in doses > or = 10 mg/day were more frequently hyperprolactinemic. CONCLUSION: Our findings that patients with SLE with a more active form of the disease and who are less responsive to therapy had increased plasma PRL levels more frequently may be indicative of a potential relationship of hyperprolactinemia to severity of disease.

The immunomodulatory properties of low-molecular tri- and tetrapeptides.

Immunomodulatory effects of six linear tri- or tetrapeptides were studied. These peptides are physiologically inert precursors, which under the action of native proteases split into a C-terminal dipeptide ester, which is subsequently converted by spontaneous intramolecular cyclization to a biologically active compound, i.e., a spirocyclic dipeptide. The following biological activities were evaluated using human lymphocytes: recovery of receptors for sheep red blood cells, test of active E-rosettes, and modulation of T-cell mitogen responses. All tested peptides revealed significant inhibition in some assay, although none of them induced significant inhibition in all assays. The compounds, I, IV, and VI proved to be the best inhibitors in comparison with Alaptide, i.e., cyclo(Ala-Acp).

Effect of domperidone-induced hyperprolactinemia on selected immune parameters in healthy women.

Domperidone, anti-emetic drug, given to healthy female volunteers, induced an elevation of plasma prolactin (PRL) concentration with the peak in 1-4 h. The release of prolactin had a transient stimulating effect on theophylline sensitive T lymphocytes and on concanavalin A induced mitogenic activity, suggesting an enhanced activity of T suppressor lymphocytes. The relative number of CD4+ lymphocytes decreased markedly one hour after domperidone administration and returned to normal values within 2 h (that means 3 h after taking the drug). The number of lymphocytes positive for dipeptidyl peptidase IV exhibited similar transient increase and normalization of activity. No change was observed in the number of CD8+ lymphocytes. The production of interferon by leukocytes treated with Newcastle disease virus was found to be significantly increased 2 h after domperidone administration. The results suggest that prolactin can selectively stimulate some functions of cellular immunity as well as the release of cytokines (IFN). The present study may contribute to the understanding of the role of the immune system in endogenous hyperprolactinemia.

Influence of various forms of dialyzable leukocyte extracts on rat adjuvant arthritis.

Adjuvant-induced arthritis in rats is a chronic inflammatory disease, widely used as an animal model for rheumatoid arthritis. In our study the effect of various fractions of dialyzable leukocyte extract (DLE): DLE I-molecular weight below 10 kDa (commercial preparation), DLE II-molecular weight below 5 kDa (suppressor fraction), DLE III-molecular weight 5-10 kDa on rat adjuvant-induced arthritis was studied. The adjuvant arthritic (AA) rats were treated with DLE fractions i.p. in solutions containing an active substance isolated from 12.5 x 10(6) and 6.25 x 10(6) leukocytes from day 1 (adjuvant injected) through day 18, every second day (total 9 times). Various markers of inflammation, immune function and joint destruction were evaluated: hindpaw volume, serum hyaluronic acid, serum albumin and biopterin in urine. All these markers showed a significant improvement after using fraction DLE II in comparison with AA controls. Fractions DLE I and DLE III influenced only some markers of inflammation and immune function. Our results demonstrated a therapeutical effect of fraction DLE II on rat adjuvant-induced arthritis.

[Determination of NK cell activity on flow cytometry]. / Stanovenie NK bunkovej aktivity na prietokovom cytometri.

The authors assessed the activity of NK cells on a flow cytometer using 5-carboxyfluorescein diacetate as fluorescent substrate. They examined 25 healthy subjects aged 23-52 years. In NK cellular activity the normal distribution applies and the value of NK cellular activity for the whole group as regards the ratio of effector cells to target cells was 25:1, 23.46 +/- 15.84 percentage. The authors did not detect any significant differences in the activity of NK cells in relation to age and sex. They revealed statistically significant correlations of NK activity and the number of CD16+ and CD57+ cells as well as CD57+ CD3- cells. The optimal ratio of effector and target cells for assessment of NK activity was 25:1. Assessment of NK activity on a flow cytometer can be considered an equivalent substitute of the classical method, using radiochromium.

Evidence for immunomodulatory properties of prolactin in selected in vitro and in vivo situations.

The relationship between neuroendocrine regulation and the immune system has recently become the subject of intense investigations. The pituitary secretes both immunostimulatory (growth hormone and prolactin) and immunosuppressive (ACTH) hormones, and is thus involved in the control of immune functions. The present work was aimed at the study of the immunoregulatory properties of prolactin in selected in vitro and in vivo model situations. Prolactin was found to enhance recovery of the receptor for sheep red blood cells (in vitro). Compared with control cells, incubation with prolactin and/or prolactin containing sera significantly enhanced the capacity of trypsin treated lymphocytes from the peripheral blood of healthy volunteers to form E-rosettes. Chlorpromazine stimulated prolactin release in males, and lactation stimulated prolactin release in females raised the number of large granular lymphocytes in peripheral circulation. Sera containing elevated prolactin levels stimulated the metabolic activity of peripheral neutrophilic leukocytes. These results suggest that prolactin may stimulate selective functions of cellular immunity, and that it is involved in interactions between the nervous, the hormonal and the immune systems.