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Despite many therapeutic regimens introduced recently, chronic lymphocytic leukemia (CLL) is still an incurable disorder. Thus, there is an urgent need to discover novel, less toxic and more effective drugs for CLL patients. In this study, we attempted to assess simvastatin, widely used as a cholesterol-lowering drug, both as a single agent and in combination with purine analogs-fludarabine and cladribine-in terms of its effect on apoptosis and DNA damage of CLL cells. The experiments were done in ex vivo short-term cell cultures of blood and bone marrow cells from newly diagnosed untreated patients. We analyzed expression of active caspase-3 and the BCL-2/BAX ratio as markers of apoptosis and the expression of phosphorylated histone H2AX (named γH2AX) and activated ATM kinase (ataxia telangiectasia mutated kinase), reporters of DNA damage. Results of our study revealed that simvastatin induced apoptosis of CLL cells concurrently with lowering of BCL-2/BAX ratio, and its pro-apoptotic effect is tumor-specific, not affecting normal lymphocytes. We observed that combinations of simvastatin+fludarabine and simvastatin+cladribine had a synergic effect in inducing apoptosis. Interestingly, the rate of apoptosis caused by simvastatin alone and in combination was independent of markers of disease progression like ZAP-70 and CD38 expression or clinical stage according to Rai classification. We have also seen an increase in γH2AX expression in parallel with activation of ATM in most of the analyzed samples. The results suggest that simvastatin can be used in the treatment of CLL patients as a single agent as well as in combination with purine analogs, being equally effective both in high-risk and good-prognosis patients. One of the mechanisms of simvastatin action is inducing DNA damage that ultimately leads to apoptosis.

Background: Sarcoidosis is a systemic granulomatous disease of unknown aetiology, in which genetic factors, especially the genes of the highly polymorphic MHC region, seem to play an important role in the disease predisposition and course. The aim of this study was to evaluate the role of TNF genes polymorphism in sarcoidosis and to estimate possible association between these polymorphisms and susceptibility and prognosis of sarcoidosis. The analysis of -308G > A TNF-alpha gene (TNFA*1 and TNFA*2 alleles) and 252A > G TNF-beta gene polymorphisms (TNFB*1 and TNFB*2 alleles) were performed., Methods: The study comprised of 130 sarcoidosis patients (75 subjects in the radiological stage I, and 55 in the stages II/III). Löfgren syndrome (LS) was manifested in 38 patients. After at least 3-years observation, 69 patients had remission, 24 subjects manifested persistent disease and 25 patients had progression. The control group consisted of 84 healthy subjects. The genotypes were determined using PCR-RFLP assay., Results: The variant allele TNFA*2 was observed significantly more frequent in patients with Löfgren syndrome when compared to control group (OR = 2.301, C.I. = [1.23-4.32], chi2 = 6.91, p > 0.01), as well as to non-LS patients (OR = 2.167, C.I. = [1.17-4.01], chi2 = 6.22, p < 0.05). Moreover, the variant allele TNFA*2 was also observed significantly more frequent in patients with disease resolution than in patients with persistent disease and progression (OR = 3.53, C.I. = [1.66-7.50], chi2 = 11.65, p < 0.001). The variant allele TNF*2 was also overrepresented in patients with disease resolution after exclusion the patients with Löfgren syndrome (OR = 2.4, C.I. = [1-5.772], chi2 = 3.98, p < 0.05). There was no significant difference in TNF-A allele distribution between the control group and whole sarcoidosis group. The variant allele TNFB*1 was observed significantly more frequent in patients with disease resolution than in patients with persistent disease and progression. This difference was caused only by overrepresentation of TNFB*1 variant allele in Löfgren group. The significant differences in the distribution of TNFB*1 allele between the sarcoidosis an the control group was also noted (OR = 1,607, C.I. = [1,033-2,5], chi2 = 4.46, p < 0.05), but it was limited only to patients displaying Löfgren syndrome., Conclusion: Two alleles TNFB*1 and TNFA*2 of TNF gene are overrepresented in polish patients with Löfgren syndrome. The TNFA*2 allele is related with mild course of sarcoidosis in patients without LS.

Chronic lymphocytic leukemia (CLL) is a heterogeneous disease with variable clinical course and prognosis. Therefore, the role of prognostic factors is very important, especially for identifying the group of patients who require intensive treatment. The aim of this study was to assess whether the rate of apoptosis caused by purine analogues differs between patients with better or worse prognostic factors. The experiments were preformed in cultures of blood and bone marrow obtained from CLL patients. The cultures were supplemented with cladribine and fludarabine. We determined the percentage of caspase-3-positive cells and the BCL-2/BAX ratio, and subsequently these apoptosis markers were correlated with the expression of ZAP-70 and CD38, lymphocyte counts, lactate dehydrogenase and beta(2)-microglobulin levels and clinical stage according to the Rai classification. The results showed that bone marrow cells are more sensitive to apoptosis caused by purine analogues than cells derived from blood, supporting the idea that these two compartments have different proliferative statuses. The cells from ZAP-70-positive patients seem to enter apoptosis more readily than those from ZAP-70-negative patients; thus, ZAP-70-positive patients are more likely to benefit from treatment with purine analogues.

B-cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are involved in normal B cell survival and differentiation. We analyzed BAFF and APRIL plasma levels in patients with B-cell chronic lymphocytic leukemia (B-CLL). We also tested intracellular BAFF and APRIL expression in B-CLL and evaluated their prognostic relevance. The percentage of leukemic B cells with intracellular APRIL or BAFF expression correlated significantly with adverse prognostic factors such as ZAP-70 and CD38. Moreover, we found a close relationship between LPL protein expression or LPL/ADAM29 MFI ratio and proportion of B-CLL cells with intracellular BAFF or APRIL expression. Furthermore, patients with a low percentage of leukemic cells with intracellular BAFF or APRIL expression had a significantly longer overall survival than those with a high proportion of APRIL or BAFF positive leukemic cells.

Introduction: Sarcoidosis is a multisystemic disease of unknown etiology characterized by the formation of immune granulomas in involved organs. The cytokine profile in inflamed lesions of sarcoidosis is mainly determined by T helper 1 (Th1) cells. Interleukin 18 (IL-18) is primarily a monocyte/macrophage-derived cytokine. IL-18 has been recently identified as an IFNgamma-inducing factor. The cytokine plays an important role in the induction of Th1 response and it may be responsible for sarcoidosis progression. The aim of the study was to assess the usefulness of IL-18 estimation in the sarcoidosis diagnosis and the disease course prognosis., Material and Methods: The diagnosis of sarcoidosis was established in 88 patients (the mean age of 38.1+/-10.8 years). We measured IL-18 level in plasma and bronchoalveolar lavage fluid (BALF) cell culture supernatant (CCS) using the enzyme-linked immunoassay technique (ELISA). We also performed the flow cytometric analysis of BALF lymphocyte phenotype. Statistica 5.0 and non-parametric tests: the Mann-Whitney U-test and the Spearman correlation test, were used for statistical analysis., Results: The patient group consisted of 55 subjects without acute symptoms of sarcoidosis, 14 patients with acute Löfgren syndrome and 19 subjects with Löfgren syndrome in the past. Lung hilar lymphadenopathy was diagnosed in 49 patients and lung interstitial changes in 39 subjects. After 6-month-observation, 49 patients were in remission, 20 subjects manifested persistent disease and 19 patients had sarcoidosis progression. Plasma IL-18 level was significantly (P<0.0001) higher in sarcoidosis patients (383+/-250pg/ml) than in control subjects (146+/-72pg/ml). Plasma IL-18 level was similar both in subjects with Löfgren syndrome and in other patients. However, IL-18 level in BALF CCS was significantly (P<0.05) lower in Löfgren syndrome patients than in subjects without acute manifestation of the disease. The highest IL-18 level in plasma was found in patients with disease progression, in subjects with lung interstitial changes and in patients with extrapulmonary manifestation of the disease. We observed a positive correlation between plasma IL-18 level and the percentage of BALF lymphocytes (R=0.202, P=0.06) as well as the percentage of activated HLA DR+T cells (R=0.23, P<0.05). There was a negative correlation between the IL-18 level in BALF CCS and the percentage of BALF CD3-positive and CD4-positive lymphocytes (R=-0.27, -0.23, P<0.05)., Conclusion: IL-18 may play a significant role in the prolongation of sarcoidosis course. Its estimation may become a good prognostic factor, which should be analyzed together with other factors useful in sarcoidosis monitoring.

Introduction: The release of tumor necrosis factor (TNF-alpha) is increased in sarcoidosis patients. TNF-alpha exerts its effect by binding to specific cell surface receptors. There are only fragmentary data concerning the expression of tumor necrosis factor receptors (TNFRs) on bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) lymphocytes. The aim of the study was to evaluate TNFRI (CD120a) and TNFRII (CD120b) expression on T cells and the level of soluble TNFRs in specimens of patients with different clinical manifestation and clinical outcome of sarcoidosis., Material and Methods: We examined 49 patients with newly diagnosed pulmonary sarcoidosis. TNFRI and TNFRII density on CD4+ and CD8+ BALF and PB cells surface was estimated using monoclonal antibodies and a flow cytometry technique. The level of TNFRs in PB serum and BALF cell culture supernatant (CCS) was measured using ELISA. Immunological analyses were also performed on PB samples collected from 10 healthy volunteers., Results: The level of soluble TNFRI (sTNFRI) in PB serum was similar in sarcoidosis patients and healthy subjects, whereas the concentration of sTNFRII in serum was significantly higher in the sarcoidosis group (P<0.001). Patients without acute symptoms of sarcoidosis, patients with radiological stage II/III as well as patients with further disease progression showed a tendency to higher levels of sTNFRs in PB serum and lower levels of sTNFRs in BALF CCS compared to Löfgren syndrome and radiological stage I subjects, and patients with spontaneous resolution of sarcoidosis. More than 80% of BALF and PB lymphocytes of sarcoidosis patients expressed both CD120a and CD120b antigens. The percentage of double-positive CD4+CD120a+ and CD4+CD120b+ cells in PB was significantly higher (P<0.005) in sarcoidosis patients than in healthy subjects. The highest percentage of CD4+CD120a+ and CD4+CD120b+ lymphocytes in BALF was determined in patients with acute disease, and in PB of patients with further spontaneous improvement., Conclusion: The evaluation of sTNFRs and TNFRs expression on T-helper cells may be useful in the estimation of sarcoidosis activity.

We describe a familial interstitial deletion of 7.7-Mb involving Xp22.2-22.3. The deletion was transmitted from an asymptomatic mother to her two children with severe developmental delay, no speech development and autistic behavior. Assessment of the deletion boundaries by FISH and PCR analyses indicated that the deletions encompasses 27 genes. Several of these genes are associated with known disorders, like KAL1 (Kallmann syndrome), steroid sulfatase (STS) (X-linked ichtyosis), and arylsulfatase E (ARSE) (chondrodysplasia punctata). The deletion also includes all four VCX genes (VCX-A, VCX-B1, VCX-B, and VCX-C) and the neuroligin 4 (NLGN4) gene. VCX-A deficiency has been shown previously to be associated with mental retardation and NLGN4 mutations lead to mental retardation in conjunction with autism. Functional deficiency of both MRX genes, VCX-A and NLGN4, are most likely associated with the impaired cognitive development of the patients described here. The phenotype associated with the Xp deletion was highly variable in female carriers and might be attributed to unfavorable X inactivation. However, all the 27 genes included in the deleted interval escape X inactivation and are expressed at variable levels from the normal X chromosome. Thus, the overall X inactivation pattern and inter-individual expression variability of the genes in distal Xp might be determinants of the phenotype associated with the deletion., (2006 Wiley-Liss, Inc.)

Background: Sarcoidosis is a multi-organ granulomatous disorder of undetermined aetiology with 90% of patients exhibiting some degree of pulmonary involvement. Transbronchial biopsy and endobronchial biopsy (EBB) with a fiber-optic bronchoscope, which demonstrate giant or epithelial cells of granuloma formation, have been established as standard methods for histological confirmation of sarcoidosis., Objectives: The analysis of EBB and bronchoalveolar lavage (BAL) specimens was used to assess the frequency of positive EBB findings in patients with pulmonary sarcoidosis. The influence of spirometric abnormalities and BAL findings on EBB-based diagnosis was also evaluated., Methods: The examined group of patients comprised 60 subjects mean age 39.4 +/- 9.65 years). The results of pulmonary function tests, EBB and BAL were analysed., Results: EBB was performed on 52 patients. Positive histological results were found in 40% of subjects; 12 of them were affected by spirometry impairment. All patients with restriction and almost half of the subjects with obstruction disorders had positive EBB findings. In the BAL fluid, the percentage of CD3+ and CD4+ cells was significantly lower and the percentage of CD19+ and CD8+ lymphocytes was significantly higher in patients with positive biopsies in comparison with the subjects with negative EBB findings., Conclusions: Positive biopsies are related to the intensity of abnormalities in the pulmonary function tests and BAL, and may reflect an advanced stage of sarcoidosis., (Copyright 2004 S. Karger AG, Basel)

HER gene family (HER1-HER4) encodes structurally similar transmembrane proteins (EGFR, HER2, ErB-3, and ErB-4) with tyrosine kinase activity. Dimerised on binding with a number of ligands, including epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha), these proteins stimulate epithelial cell proliferation. HER2 and EGFR overexpression is detected in the cells of many tumours, mainly in breast, lung and oral cancer and may be connected with HER2 gene amplification or point mutations as well as with the presence of overactive polymorphic forms of HER1 gene. The first medication of a proved efficacy in breast cancer treatment was trastuzumab (Herceptin)--monoclonal antibody against HER2 protein. Trastuzumab was effective only in the case of patients with high HER2 expression evaluated by immunohistochemical methods and with gene amplification ascertained by fluorescence in situ hybridisation assays. In non-small-cell lung cancer (NSCLC), HER2 overexpression was detected only in a few cases. Therefore, trastuzumab treatment seems to be problematic in NSCLC patients. A small molecule quinazoline (erlotinib, Tarceva) is a promising therapeutic agent selectively blocking EGFR. Phase III Tarceva clinical trail in NSCLC patients showed that their survival is prolonged and that the medication acts together with other chemotherapeutic agents like cisplatin and gemcitabine.

Dendritic cells play a specific regulatory role in the immune system. In this paper, the significance of myeloid and lymphoid dendritic cells in sarcoidosis and extrinsic allergic alveolitis (EAA) was evaluated. Myeloid dendritic cells are connected with Th1 type of immunological response, whereas lymphoid ones--with Th2 type. The latest findings indicate that both diseases are characterized by serious disturbances of Th1/Th2 response to Th1 dominance. Our studies seem to confirm these suggestions. In the peripheral blood of patients with sarcoidosis as well as with EAA, myeloid dendritic cells outnumbered lymphoid ones.

The objective of the research was to evaluate immunological and clinical parameters in bronchial asthma patients from rural and urban areas in the east part of Poland. The study comprised 55 subjects reporting from November 2001 to January 2002 at the Pulmonary Department in order to have the disease diagnosed or to continue the treatment. The type of symptoms was determined on the basis of the standardized questionnaire. The patients were subjected to spirometry and skin prick tests. Blood eosinophil number as well as total and specific IgE levels were also evaluated. Significant (p<0.05) disturbances in respiratory parameters were found in 23% of patients living in the city and in 50% of subjects from the rural area. The level of IgE against grass pollens was significantly (p<0.01) higher in urban residents. The level of IgE against house dust was similar in both groups. Our study shows important differences between clinical manifestation of asthma in rural and inner-city patients and indicates that different aetiology may play an important role in asthma development.