Bogdan Mazur

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.

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.

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 ΔΨm 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.

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.

Staphylococcus aureus skin colonization in atopic dermatitis children is associated with decreased IFN-γ production by peripheral blood CD4+ and CD8+ T cells

Atopic dermatitis (AD) is a chronic inflammatory skin disorder, which is associated with an increased expression of Th2 cytokines with concomitant decrease in IFN‐γ production by circulating CD4+ and CD8+ T cells. The skin of patients with AD is often colonized by Staphylococcus aureus, which may reflect in changes in immunological parameters. The aim of the study was flow cytometric measurement of some peripheral blood lymphocyte subsets expressing naive/memory marker (RA/RO) and activation marker (CD25) as well as intracellular production of IFN‐γ by peripheral blood CD4+ and CD8+ T cells from varied severity AD children and determine the impact of S. aureus skin colonization on cytokines profiles. There was 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+ and CD8+ T cells producing IFN‐γ upon in vitro stimulation with phorbol 12‐myristate 13‐acetate and ionomycin in children with AD compared to healthy ones. The absolute number of CD4+ and CD8+ T cells expressing memory marker CD45RO was elevated as compared with controls. The severity of AD was positively correlated with the percentage of lymphocyte subsets: CD45RO+, CD4+CD45RO+, and the percentage of CD3+ and CD4+ expressing CD25 as well as the number of S. aureus on the skin. In conclusion, both CD4+ and CD8+ memory T cells are involved in the immunopathogenesis of AD. S. aureus skin colonization is related with disease severity and changes in expression of CD45RO and CD25 on T cells. A decrease in the percentage of CD4+ and CD8+ T cells producing IFN‐γ in AD children may explain propensity for skin infection.

Analysis of procalcitonin and CRP concentrations in serum of patients with chronic spontaneous urticaria

BackgroundOur previous findings showed the importance of analysing the peripheral markers of acute phase response (APR) activation, C-reactive protein (CRP) and IL-6 in the context of urticaria activity and severity. However, these biomarkers do not reliably differentiate between APR to infectious and the disease severity.AimIn order to investigate a possible association between the immune-inflammatory activation markers CRP and procalcitonin (PCT).MethodsSerum PCT and CRP concentrations were measured in patients with CU of varying severity as well as in healthy subjects.ResultsSerum PCT and CRP concentrations were significantly increased in more severe CU patients when compared to healthy controls and mild CU, and within the CU population there was a significant correlation between concentrations of PCT and CRP. Serum PCT concentrations remained within normal ranges in most CU patients and were only slightly elevated in some severe CU cases.ConclusionsPCT serum concentration may be only slightly elevated in some cases of severe CU. Upregulation of PCT synthesis accompanied by parallel changes in CRP concentration reflects a low-grade systemic inflammatory response in CU. PCT should be considered as a better marker than CRP to distinguish between APR to infection and an active non-specific urticarial inflammation.

The Immunophenotypes of Blast Cells in B-Cell Precursor Acute Lymphoblastic Leukemia: How Different Are They from Their Normal Counterparts?

Currently, there are three major maturational stages of CD19 antigen expressing B‐cell precursors (hematogones). In B‐cell precursor acute lymphoblastic leukemia (BCP‐ALL), the malignant counterpart of hematogones, the leukemic blasts share common phenotypic features. The aim of the study was to enumerate the actual differences between the leukemic blasts in the CD10+ and CD10− subgroups of BCP‐ALL and hematogones by assessing the expression of the antigens: TdT, CD34, CD45, CD10, CD38, CD20 and CD22.

Increased serum complement C3 and C4 concentrations and their relation to severity of chronic spontaneous urticaria and CRP concentration

Chronic spontaneous urticaria (CU) is associated with activation of the acute phase response (APR). Nevertheless, APR-associated proteins have not been well characterized as potential biomarkers of the disease severity. To assess the pattern of complement proteins C3 and C4 – the acute phase reactants in patients with CU. C3, C4 and CRP concentrations were measured in serum of 70 patients showing different degrees of urticarial severity as well as in 33 healthy subjects. Serum C3 and C4 concentrations were significantly increased in CU patients as compared with the healthy subjects and exceed the normal lab range by about 5% and 10%, respectively. Significant differences were found between patients with mild and increased CU severity. In addition, significant correlations were observed between C3, C4 and CRP concentrations. More severe CU is characterized by higher production of C3 and C4 complements accompanied by parallel changes in CRP concentration.

Clinical Features and Immunological Markers of Atopic Dermatitis in Elderly Patients

Background: Atopic dermatitis (AD) is increasing among the elderly. Staphylococcus aureus colonization is one of the most important aggravating factors in AD. Objective: This study analyses the clinical features of AD in elderly patients and determines the pathogenic roles of staphylococcal superantigens in AD with low and high total IgE levels. Methods:S. aureus enterotoxin genes were evaluated using PCR. Additionally, S. aureus-specific IgE levels and peripheral blood lymphocyte profiles were assessed. Results: A total of 44 women and 77 men diagnosed with AD with a mean age of 68.92 ± 6.51 years were evaluated. In 17 (68%) patients with AD and low levels of total IgE, there was a positive correlation between the positive results for enterotoxin B, S. aureus-specific IgE antibodies and Th1 cytokine profiles (Spearman’s rank correlation test, r = 0.89, p < 0.05). Conclusion: Our results indicate that AD patients with low total IgE levels differ in immunopathogenesis from AD patients with high circulating levels of total IgE AD.

LYMPHOCYTE SUBPOPULATIONS IN FULL-TERM SEPTIC NEONATES

Abstract Purpose: To estimate the absolute leukocyte and lymphocyte counts and relative and absolute sizes of CD19+ B lymphocytes, CD3+, CD4+, CD8+ and CD3+/HLA–DR+ T lymphocytes in full‐term septic neonates and the influence of some perinatal risk factors on these lymphocyte subsets.