Multicolor flow cytometry immunophenotyping and characterization of aneuploidy in pediatric B-cell precursor acute lymphoblastic leukemia.

The aim of this study was to assess the incidence of DNA aneuploidy in Polish children with B-cell precursor acute lymphoblastic leukemia (BCP-ALL) and the relationship between aneuploidy and immunological phenotype, age, leukocyte count, S-phase fraction (SPF) and early response to induction chemotherapy assessed by the percentage of residual blast cells in bone marrow aspirates. The study group consisted of 267 patients. DNA content and immunophenotype were assessed in the bone marrow before treatment using multicolor flow cytometry (FC). DNA aneuploidy was detected in 50/267 (19%) patients. High hyperdiploidy was found to be associated with lower leukocyte count (p = 0.006) and common ALL immunophenotype. Flow cytometry analysis revealed that high hyperdiploid BCP-ALL patients showed significantly higher expression of CD9, CD20, CD22, CD58, CD66c, CD86 and CD123 antigens as compared to other groups of ploidy. In contrast, CD45 showed decreased expression. The percentage of leukemic blasts at diagnosis was lower in high hyperdiploid BCP-ALL cases than in diploid (79% vs. 85.7%, p = 0.001). The difference in minimal residual disease (MRD) levels on day 15 and 33 of induction therapy between analyzed groups was not significant. This study showed that high hyperdiploidy is associated with lower WBC count and specific immunological phenotype. Flow cytometric evaluation of expression of selected antigens can be used for fast identification of markers of aneuploidy in pediatric BCP-ALL, before genetic tests results are available. Understanding the biological significance of aneuploidy in leukemia can potentially be exploited therapeutically using targeted therapies against specific blast cell subclones.

Novel interleukin-33 and its soluble ST2 receptor as potential serum biomarkers in parotid gland tumors.

An increasing number of patients with parotid gland tumors have been observed in recent years. The relationship between the immune system and tumor formation is thoroughly investigated. However, newly discovered molecules offer a new insight into the pathophysiology of malignancies. It would be ideal to find an easily determinable biomarker of tumor existence, its malignant potential or a biomarker suggesting the probability of disease recurrence. Our study is the first to examine serum concentrations of IL-33 and its sST2 receptor in patients with various types of parotid gland tumors. Serum IL33, sST2, IL-4 and IL-10 concentrations were determined in patients with benign and malignant parotid gland tumors (pleomorphic adenoma, Warthin's tumor, myoepithelioma and acinic cell carcinoma). We observed for the first time that serum IL-33 level was significantly elevated in patients with various types of parotid gland tumors and sST2 levels were significantly higher in pleomorphic adenoma and acinic cell carcinoma patients compared to the controls. Our results demonstrate for the first time that serum IL-33 and its sST2 receptor may be important factors in the pathology of parotid gland tumors. Although our results are promising, further investigations are required to detect if serum concentrations of those molecules may be a biomarker in parotid gland tumors. Impact statement Parotid gland tumors seem to be an increasingly important medical challenge, mostly due to a noticeable increase in the incidence. It would be crucial to find an easily determinable biomarker of tumor existence, its recurrence or malignant potential. We observed for the first time that serum IL-33 level was significantly elevated in patients with various types of parotid gland tumors and its sST2 receptor levels were significantly higher in pleomorphic adenoma and acinic cell carcinoma patients compared to the controls. We believe that our study helps to understand the biology of the tumors and a potential role of a relatively newly identified cytokine IL-33 in the pathophysiology of the parotid gland tumors.

ATP4A autoimmunity in pediatric patients with type 1 diabetes and its relationship to blood count, iron metabolism, and vitamin B12.

OBJECTIVE: We aimed to assess the prevalence of autoantibodies against the 4A subunit of the gastric proton pump (ATP4A) in pediatric type 1 diabetes (T1D) patients and explore the relationship between ATP4A positivity and blood cell count, iron turnover, and vitamin B12 concentration. SUBJECTS: The study included 94 (59% female) T1D children (aged 12.5 ± 4.1 years, T1D duration 4.2 ± 3.6 years, HbA1c 7.3 ± 1.5% (57 ± 12.6 mmol/mol) with no other autoimmune diseases. METHODS: ATP4A antibodies were measured in T1D patients using a radioimmunoprecipitation assay. Blood cell count, iron concentration, total iron binding capacity, ferritin, transferrin, hepcidin, and vitamin B12 concentration were measured in all the study participants. RESULTS: A total of 16 (17%) children were ATP4A positive. Serum concentrations of ferritin were significantly lower in ATP4A positive than in antibody negative subjects (P = .034). Overall the levels of ATP4A antibodies (ATP4A Index) correlated positively with the age at T1D diagnosis (r = 0.228, P = .026) and negatively with ferritin levels (r = -0.215, P = .037). In ATP4A positive patients, the ATP4A Index correlated positively with age at diagnosis (r = 0.544, P = .032) and negatively with vitamin B12 levels (r = -0.685, P = .004). CONCLUSIONS: ATP4A antibodies were present in a significant proportion of children with T1D. Higher ATP4A levels in T1D children are associated with lower, yet still fitting within the normal range, levels of vitamin B12, and ferritin. Routine screening of T1D children for gastric autoimmunity (ATP4A) should be considered with follow-up of those positive for vitamin B12 and iron deficiency.

The inhibitory effect of flavonoids on interleukin-8 release by human gastric adenocarcinoma (AGS) cells infected with cag PAI (+) Helicobacter pylori.

INTRODUCTION: It is well known that the presence of Helicobacter pylori in the stomach induces gastritis and causes an immune response. Exposure of gastric epithelial cell lines to this germ induces the secretion of interleukin-8 (IL-8), which is a potent PMN-activating chemotactic cytokine. Interleukin-8 is usually elevated in gastric biopsy samples of patients with H. pylori-associated gastritis and significantly increases in the supernatant of in vitro cultivated biopsy samples of gastric mucosa with active H. pylori gastritis. Interleukin-8 is an activating factor for leucocytes and other pro-inflammatory factors, free radicals, and proteolytic enzymes. That is why natural compounds potentially useful in therapy are still investigated - among them flavonoids. They reveal anti-oxidative and anti-inflammatory activities and significantly inhibit the gastric mucosa damage. THE AIM OF THE STUDY: Was the estimation of the anti-inflammatory effects of flavonoids on H. pylori-induced activation of human gastric adenocarcinoma cells (AGS). After infection of AGS cells by cag PAI (+) H. pylori in vitro, secretion of IL-8, effects of flavonoids on viability of AGS cells, and effects of flavonoids on increase of H. pylori were determined. Such flavones as chrysin, quercetin, kaemferide, flavanone, galangin, and kaempferol were examined. RESULTS: This study has shown an inhibitory effect of flavonoids on the release of IL-8 through infected AGS cells (except chrysin), and no toxic effects to AGS cells were observed. Galangin revealed antibacterial effects against H. pylori. Flavonoids limit the inflammatory process through the inhibition of IL-8 release in infected AGS cells with H. pylori. The strongest inhibitor of IL-8 was galangin.

Assessment of selected B cells populations in the workers of X-ray departments.

OBJECTIVES: Workers of X-ray departments are occupationally exposed to long-term low levels of ionizing radiation (LLIR), which may affect their humoral immunity. The aim of the study was to assess the influence of LLIR on the number and proportion of B cells (CD19+), B1 cells (CD5+CD19+) and memory B cells (CD27+CD19+) in peripheral blood of such workers. MATERIALS AND METHODS: In the study group of 47 X-ray departments workers and the control group consisting of 38 persons, the number and percentage of CD19+, CD5+CD19+, CD27+CD19+ cells as well as CD5+CD19+/CD19+ and CD27+CD19+/CD19+ cell ratios were assessed using flow cytometry. Additionally, the study group was divided into 2 groups by the length of employment below and over 15 years and analysis adjusted for age and smoking habit was performed. RESULTS: The total number of CD19+ cells showed significant increase in the group of workers in comparison with the persons from the control group, whereas the percentage of CD5+CD19+ cells as well as CD27+CD19+/CD19+ and CD5+CD19+/CD19+ cell ratios were lower. Percentage, number of CD5+CD19+ cells and CD5+CD19+/CD19+ cell ratio were significantly lower in the workers with length of employment longer than 15 years in comparison with those employed below 15 years. Moreover, we found positive associations between the number of CD19+ cells and employment as well as smoking habit, whereas the number of CD5+CD19+ cells was positively associated with cigarette smoking alone. Percentage of CD5+CD19+ cells as well as CD5+CD19+/CD19+ and CD27+CD19+/CD19+ cell ratios were negatively correlated with employment. CONCLUSIONS: The study suggests association between the suppressive influence of low level ionizing radiation on circulating in peripheral blood, especially of B1 cells as well as of memory B cells, in workers of X-ray units, which is adverse in relation to microbiological threat.

Chalcones and dihydrochalcones augment TRAIL-mediated apoptosis in prostate cancer cells.

Chalcones and dihydrochalcones exhibit chemopreventive and antitumor activity. TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a natural endogenous anticancer agent. We examined the cytotoxic and apoptotic effect of chalcones and dihydrochalcones on TRAIL-mediated apoptosis in LNCaP prostate cancer cells. The cytotoxicity was evaluated by the MTT and LDH assays. The apoptosis was detected using annexin V-FITC by flow cytometry and fluorescence microscopy. The DeltaPsim was evaluated using DePsipher staining by fluorescence microscopy. Our study showed that two tested chalcones (chalcone and 2',6'dihydroxy-4'-methoxychalcone) and three dihydrochalcones (2',6'-dihydroxy-4'4-dimethoxydihydrochalcone, 2',6'-dihydroxy-4'-methoxydihydro- chalcone, and 2',4',6'-trihydroxydihydrochalcone, called phloretin) markedly augmented TRAIL-induced apoptosis and cytotoxicity in LNCaP cells and confirmed the significant role of chalcones in chemoprevention of prostate cancer.

Cytokine production by peripheral blood CD4+ and CD8+ T cells in atopic childhood asthma.

There are conflicting studies on T cell cytokine production in childhood asthma. In this study intracellular cytokine expression of IL-2, IL-4, IL-10, IL-13, IFN-γ, and TNF-α in CD4(+) and CD8(+) T cells in children with atopic asthma were measured by flow cytometry. Results. A significant increase in the percentage of CD4(+) and CD8(+) T cells producing IL-4 and IL-13 and decrease in the percentage of CD4(+) producing IFN-γ in asthmatic children was found. The percentage of CD4(+)/IL-13(+) was significantly higher in severe asthma than in children with intermittent disease symptoms. Severity of asthma was associated with increased both serum IgE and frequencies of CD4(+)/IL-13(+) T cells, as well as duration of disease. Moreover, a decrease in FEV(1), FEV(1)/FVC was observed in relation to the severity of asthma. Changes in cytokine profile in CD8(+) subpopulation didn't depend on the severity of the disease. Conclusions. Increased production of IL-4 and IL-13 in both CD4(+) and CD8(+) T cells accompanied by decreased IFN-γ expression in CD4(+) T cells may be evidence that both lymphocyte subpopulations are implicated in the pathogenesis of asthma. Relationship of CD4(+)/IL-13(+) T cells with disease activity suggests that this lymphocyte subset may have a prominent role in childhood asthma.

Ethanolic extract of propolis (EEP) enhances the apoptosis- inducing potential of TRAIL in cancer cells.

Ethanolic extract of propolis (EEP) is one of the richest sources of phenolic acids and flavonoids. EEP and its phenolic compounds have been known for various biological activities including immunopotentiation, chemopreventive and antitumor effects. Tumor necrosis factor related apoptosis inducing ligand (TRAIL) is a naturally occurring anticancer agent that preferentially induces apoptosis in cancer cells and is not toxic toward normal cells. We examined the cytotoxic and apoptotic effect of EEP and phenolic compounds identified in propolis in combination with TRAIL on HeLa cancer cells. HeLa cells were resistant to TRAIL-induced apoptosis. Our study demonstrated that EEP and its components significantly sensitize to TRAIL induced death in cancer cells. The percentage of the apoptotic cell after exposure to 50 microg/mL EEP and 100 ng/mL TRAIL increased to 71.10 +/- 1.16%. The strongest cytotoxic effect in combination with TRAIL on HeLa cells exhibited apigenin and CAPE at the concentration of 50 microM (58.87 +/- 0.75% and 49.59 +/- 0.39%, respectively). In this report, we show for the first time that EEP markedly augmented TRAIL mediated apoptosis in cancer cells and confirmed the importance of propolis in chemoprevention of malignant tumors.

TRAIL-induced apoptosis and expression of death receptor TRAIL-R1 and TRAIL-R2 in bladder cancer cells.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) is a member of TNF superfamily able to induce programmed death in cancer cells with no toxicity against normal tissues. TRAIL mediate apoptosis follows binding to the two death receptors, TRAIL-R1 (DR4) and/or TRAIL-R2 (DR5). In this study we investigated the cytotoxic and apoptotic effect of TRAIL on bladder cancer cells and the expression of death receptor TRAIL-R1 and TRAIL-R2 on the surface of these cancer cells. Three human bladder transitional cancer cell (TCC) lines - SW780, 647V and T24 were tested for TRAIL sensitivity. The bladder cancer cells were incubated with human soluble recombinant TRAIL. Cytotoxicity was measured by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-dimethyltetrazolium bromide) and LDH (lactate dyhydrogenase) assays. Apoptosis was detected by flow cytometry with annexin V-FITC/propidium iodide and by fluorescence microscopy with Hoechst 33342/annexin V-FITC/Ethidium Homodimer. The cell surface expression of TRAIL death receptors on bladder cancer were determined using flow cytometry with phycoerythrin-conjugated monoclonal anti-human TRAIL-R1 and TRAIL-R2. Our investigations confirmed that SW780 cells were sensitive to TRAIL, and two other bladder cancer cell lines, 647V and T24, were resistant to TRAIL induced apoptosis. We therefore examined the expression of TRAIL death receptors on bladder cancer cell surfaces. We showed decreased expression of TRAIL-R2 receptor in TRAIL-resistant bladder cancer cells and increased expression of this death receptor in TRAIL-sensitive SW780 cells. The expression of TRAILR1 receptor was similar in all bladder cancer cell lines. TRAIL is one of the promising candidates for cancer therapeutics. However, some cancer cells are resistant to TRAIL-mediated apoptosis. It is therefore important to overcome this resistance for the clinical use of TRAIL in cancer therapy. TRAIL death receptors are attractive therapeutic targets in cancer treatment. The cytotoxic agents capable of up-regulating the expression of TRAIL-R1 and TRAIL-R2 can sensitize cancer cells to TRAIL induced apoptosis.

Chalcones enhance TRAIL-induced apoptosis in prostate cancer cells.

Chalcones exhibit chemopreventive and antitumor effects. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a naturally occurring anticancer agent that induces apoptosis in cancer cells and is not toxic to normal cells. We examined the cytotoxic and apoptotic effect of five chalcones in combination with TRAIL on prostate cancer cells. The cytotoxicity was evaluated by the MTT and LDH assays. The apoptosis was determined using flow cytometry with annexin V-FITC. Our study showed that all five tested chalcones: chalcone, licochalcone-A, isobavachalcone, xanthohumol, butein markedly augmented TRAIL-mediated apoptosis and cytotoxicity in prostate cancer cells and confirmed the significant role of chalcones in chemoprevention of prostate cancer.

Erythrocyte antioxidant parameters in workers occupationally exposed to low levels of ionizing radiation.

It has been postulated that ionizing radiation generates reactive oxygen species (ROS). ROS are annihilated by an intracellular enzymatic system composed mainly of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Workers of X-ray departments are occupationally exposed to long-term low levels of ionizing radiation, which may affect their antioxidant status. Erythrocyte activities of SOD, CAT and GPx were measured in 45 workers of X-ray departments and 30 persons who constituted the control group. Subgroups with respect to sex and cigarette smoking were selected. Colorimetric method was used for determination erythrocyte activities of SOD, CAT and GPx. A significant decrease of GPx, SOD and CAT activity in workers as compared to controls was observed. Lower activity of SOD and GPx in female and GPx in male subgroup was found. SOD was significantly more elevated in smoking workers than in the non-smoking staff. Moreover non-smoking employees showed lower SOD and GPx activity in comparison to the non-smoking control. GPx decrease was found in smoking workers in comparison to the smoking control. Additionally, smoking workers showed lower activity of GPx and CAT compared to non-smoking control.

[Comparative analysis of blast immunophenotype between the diagnosis and relapse of childhood acute lymphoblastic leukaemia - implications for monitoring of minimal residual disease]. / Analiza zmian immunofenotypu komorek bialaczkowych przy diagnozie oraz wznowie w dzieciecej ostrej bialaczce limfoblastycznej oraz ich wplywu na wykrywanie minimalnej choroby resztkowej.

PURPOSE: The aim of the present study was to compare the blast immunophenotype of acute lymphoblastic leukaemia (ALL) at diagnosis and at relapse and to define the most frequent shifts in marker expression. PATIENTS AND METHODS: Bone marrow samples from 14 patients were analyzed by flow cytometry both at diagnosis and at relapse - in 12 patients with B-cell precursor (BCP)-ALL and in 2 patients with T-ALL. RESULTS: The conversion in blast immunophenotype was observed in 12 out of 14 patients (86%). Antigen CD34 turned out to be the most unstable antigen - the shift in the signal expression was present in 57% of BCP-ALL and in both T-ALL cases. Regarding B-lineage markers, the shifts most frequently concerned CD20 (shifts present in 41.5% of cases) and CD22 (27%). Among the T-lineage markers: CD3, CD4 and CD8 demonstrated the highest incidence of altered signal expression. On the other hand, the most stable antigen included CD19 and CD10 for the BCP-ALL group and CD1a, CD2, CD5, CD7 for T-ALL patients. Expression of HLA-DR, TdT and CD45 antigens remained unchanged in both BCP-ALL and T-ALL groups. CONCLUSIONS: The results of the present study support the requirement to monitor at least two different leukaemia specific antigen combinations for detection of MRD to prevent a false-negative result and to increase the effectiveness of monitoring minimal residual disease.

CD30 on stimulated CD4+ T lymphocytes in newborns regarding atopic heredity.

CD30 was initially described as Ki-1 Ag on Reed-Sternberg cells of Hodgkin's lymphoma and its and CD30L(+) expression on T cells in placenta were equally frequent in the atopic and non-atopic women. In this article we present a study of CD30 mean fluorescence intensity (MFI) on CB T CD4(+) cells. We tested the hypothesis that in newborns with atopy family history there is a changed CB T cells response after antigen stimulation comparing with those without atopy family history. The study population consisted of 31 newborn babies (29-breastfed, two non-breastfed) and their mothers. Eleven of them had positive and 20 had negative atopy family history. Performed tests included cord blood, which was a subject to flowcytometry analysis and was cultured for 24 h, cytokine production was measured (IFN- gamma, IL-4 and IL-12). Secondly, we measured total maternal and cord blood IgE levels. We studied CD30 MFI as in our studies in larger group of newborns, CD30 expression on CD4(+) T cells appeared to be very low. MFI of CD4(+) CD30(+) after PHA-stimulation (213.55: range: 41.77-434.51) was significantly increased compared to MFI of CD4(+) CD30(+) before PHA-stimulation (43.63: range 28.67-134.67)(p 0.05). After PHA stimulation MFI of CD4(+) CD30(+) in non-atopic (273.05 (range: 42.9-434.51) was significantly increased compared with the atopic newborns to MFI of 87.1 (range: 41.78-241.42) (p = 0.00). We have not found any correlation between MFI of CD4(+) CD30(+) and total maternal and total CB IgE levels. The role of CD30 in immunological response needs further research studies.

Plasma ghrelin levels in children with chronic renal failure on peritoneal dialysis.

OBJECTIVES: Malnutrition and loss of appetite represent a serious problem in children with chronic renal failure. Ghrelin is a newly described hormone involved in control of growth hormone secretion, stimulation of food intake, and regulation of energy balance. METHODS: Plasma ghrelin levels were compared between 12 children on automated peritoneal dialysis (APD) and 9 children on conservative treatment of chronic renal failure. Eight healthy children matched for age and body mass index (BMI) served as a control group. RESULTS: Plasma ghrelin levels were similar in children on APD (698.3 +/- 59.7 pg/mL) and children on conservative treatment (675.4 +/- 41.9 pg/mL) compared to healthy controls (700.1 +/- 24.7 pg/mL). There was no difference in plasma ghrelin levels in children with chronic renal failure regardless of the method of treatment (peritoneal dialysis vs conservative treatment). The plasma ghrelin index was similar in all three investigated groups: APD 40.2 +/- 8.7 vs conservative treatment 39.1 +/- 5.6 vs controls 41.0 +/- 7.8 (pg/mL)/BMI (kg/m2). Plasma ghrelin levels did not correlate with age, duration of dialysis treatment, height, weight, BMI, creatinine and urea levels, adequacy parameters, or nightly glucose load. CONCLUSION: Plasma ghrelin levels in children on APD were not different from levels in children on conservative treatment or healthy controls with comparable BMI. The persistent state of toxic influence of uremic end-products could be responsible for such a lack of correlation with anthropometrical parameters. Further studies on a larger group of children on APD are needed to clarify the effect of ghrelin on nutritional status in children with chronic renal failure.

Intracellular production of IL-2, IL-4, IFN-gamma, and TNF-alpha by peripheral blood CD3+ and CD4+ T cells in children with atopic dermatitis.

The role of the type-2 T helper (Th2) cell-mediated immune response in the immunopathogenesis of atopic dermatitis (AD) is well documented. Whether polarized immunoresponse is confined to antigen-specific T cells or is distributed among all T cell subsets is still controversial. We investigated frequencies of interleukin-2 (IL-2), IL-4, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) producing CD3(+) and CD4(+) T cells in peripheral blood from children with atopic dermatitis and healthy subjects with and without in vitro stimulation. Children with severe AD had a significantly lower percentage of CD4(+) T cells spontaneously expressing IL-4 compared with healthy controls (p <0.01). Polyclonal stimulation significantly increased cytokine production in both AD patients and healthy individuals. Frequencies of CD3(+) and CD4(+) producing IL-2, IL-4, IFN-gamma, and TNF-alpha after in vitro stimulation with phorbol-12-myristate 13-acetate (PMA) + ionomycin were comparable in the AD and control groups. In response to PMA/ionomycin, children with AD and asthma symptoms had a significantly lower percentage of CD3(+) T cells producing TNF-alpha. We failed to demonstrate evidence of an imbalance with respect to type-2 cytokine productions in children with AD. Comparable induction of Th1 and Th2 cytokines in polyclonally stimulated peripheral CD3(+) and CD4(+)T cells from AD patients and controls puts into question the polarized Th2 immune response as a general characteristic of T cells in children with atopic dermatitis.

Expression of cellular isoform of prion protein on the surface of peripheral blood lymphocytes among women exposed to low doses of ionizing radiation.

Ionizing radiation affects the expression of adhesive and co-stimulatory molecules in lymphocytes. The physiological function of cellular isoform of prion protein (PrPc) is little known. Evidences indicate a link between lymphocytes activation and PrPc expression on their surface; however, no direct effect of radiation on PrPc level in these cells was investigated. The objective of this study was to determinate the effect of low doses of ionizing radiation on the expression of PrPc on the surface peripheral blood lymphocytes in the women operating X-ray equipment. In 36 female workers and 30 persons of the control group the PrPc expression on CD3 (T lymphocytes), CD4 (T helper), CD8 (T cytotoxic) and CD19 (B lymphocytes), as well as the percentage of lymphocytes with PrPc on their surface, were tested. Subgroups with respect to age and length of employment were selected. A significant increase was observed in PrPc expression on CD3 and CD4 with lowered PrPc level on CD8 and percentage of CD8 cells with PrPc in workers compared to control. The PrPc level did not show significant changes in subgroups in relation to age (below and over 40 years old) both in the investigated and control groups, whereas a lower percentage of PrPc expressing CD19 cells showed in employed women below 40 years of age. A significant decrease was found in PrPc expression on the surface of CD3, CD4 and CD8 cells in the subgroup employed for over 10 years than in the subgroup with less than 10 years of employment.

Plasma ghrelin concentrations in patients with polycystic ovary syndrome before and after 6months therapy: correlation with androgen levels.

OBJECTIVE: Ghrelin is a natural ligand of the growth hormone secretagogue receptor and has been shown to be a potent stimulant of GH secretion. It has also orexigenic effects and regulates energy homeostasis. Recent studies claim that ghrelin influences the androgen level and probably takes part in PCOS pathomechanism. The aim of the study was an assessment of ghrelin level in plasma in women with PCOS before and after the treatment and ghrelin's influence on androgen level change. MATERIAL AND METHODS: The study included 25 women with the diagnosed PCOS (mean age 25.3+/-4.05 yr). The tests were done twice: before the treatment and after 6-month therapy with Diane 35 (cyproterone acetate 2 mg with ethinylestradiol 35 mug). Following hormones were measured: ghrelin, free testosterone, androstendione, dehydroepiandrosterone sulfate, 17-OH-progesterone and estradiol. RESULTS: The received results in both groups were compared with the control group (11 healthy women, mean age 26.0+/-2.6 yr). No statistically significant differences in ghrelin levels before (187.8+/-8.1 fmol/ml) and after the therapy (185.6+/-9.5 fmol/ml) were found. Similar results were received when two groups of women compared with the control (186.5+/-8.7 fmol/ml). No correlations between ghrelin and androgen levels were confirmed. CONCLUSIONS: Final conclusion is that there is no direct impact of ghrelin level on PCOS pathogenesis, however, its role in development of obesity, hyperinsulinemia and insulin resistance co-occurring with metabolic disorders syndrome cannot be excluded.

Decreased numbers of CD4+ T lymphocytes in peripheral blood after treatment of childhood acute lymphoblastic leukemia.

Treatment-related immunosuppression in patients with acute lymphoblastic leukemia (ALL) is associated with increased susceptibility to infectious diseases, also after the treatment. The aim of the present study was the detailed evaluation of T lymphocyte subsets in peripheral blood in children after treatment of ALL. All children were treated according to the BFM 90 protocol. The patients were divided into 5 groups of 30 children in each, depending on the time from cessation of the ALL treatment. A control group consisted of 30 healthy children subjected to elective "1-day" surgery. The children's age ranged from 6 to 18 years. The examinations were performed in FACScan flow cytometer with the use of wide set of monoclonal antibodies: CD3, CD4, CD8, TCRalphabeta, TCRgammadelta, CD19, CD25, CD45RA, CD45RO, CD69, HLA-DR, CD16 and CD56, which particularly allowed detailed analysis of T lymphocytes. The results showed that most parameters in children 1 year after ALL treatment completion were similar to healthy children. However, we observed persistently low CD4+ T cell numbers, both CD45RA+ as well as CD45RO+ subsets as compared to the control group. This might reflect decreased regenerative potential of immunological system in children 1 year after ALL treatment.

Assessment of blood serum immunoglobulin and C-reactive protein concentrations in workers of x-ray diagnostics units.

OBJECTIVES: Workers of x-ray departments are occupationally exposed to long-term low levels of ionizing radiation. The aim of the study was to investigate the influence of occupational exposure of low-level x-ray radiation on immunoglobulin and C-reactive protein (CRP) concentrations in radiology workers. MATERIALS AND METHODS: In the study group of 41 x-ray department workers and the control group composed of 32 persons, immunoglobulins (IgM, IgG, IgA) and CRP concentrations were analyzed. The study group was subdivided by gender and smoking habit. RESULTS: A significant decrease in IgG level was found in the workers and the female subgroup. The same observation was made when smokers and nonsmokers of both groups were compared. Smoking workers showed lower concentrations of IgA than non-smokers. The remaining results of immunoglobulin and CRP concentrations did not show significant differences. CONCLUSIONS: Occupational exposure to low levels of ionizing radiation is associated with suppressive influence on the immunoglobulin production, especially IgG. In addition, smoking decreases the production of IgA in radiology workers.

Concentration of IL-2, IL-6, IL-8, IL-10 and TNF-alpha in children with acute lymphoblastic leukemia after cessation of chemotherapy.

The immunosuppressive effect of cytotoxic drugs, basic therapeutic agents in the treatment of childhood acute leukemias, requires monitoring of the immune system following cessation of therapy. The cytokines are soluble proteins that play a key role in the immunoregulation of the lymphocyte function. The cytokines regulate growth, differentiation and function of various cells in normal conditions. The aim of our study was to estimate serum levels of IL-2, IL-6, IL-8, IL-10 and TNF-alpha in children with acute lymphoblastic leukemia (ALL) after cessation of chemotherapy. The study involved 150 children with ALL. This group consisted of: 30 children 1 month after treatment cessation; 30 children, 3 months later; 30 children 6 months later; 30 children, 9 months later and 30 children, 12 months later. The control group consisted of 30 healthy children. The levels of the cytokines under study were assayed using the immunoassay kits (R&D Systems, USA). During the study significant differences in TNF-alpha, IL-2 and IL-8 serum concentrations were observed among treated children and controls. However there were no differences in IL-6 and IL-10 concentrations.