Vitamin D analogs decrease in vitro secretion of RANTES and enhance the effect of budesonide.

PURPOSE: Eosinophils appear to be central inflammatory cells in the pathogenesis of rhinosinusitis with nasal polyps (NP). One of the most predominantly recognized eosinophil chemoattractants is RANTES. The aim of this study was to assess the influence of vitamin D (VD) derivates on RANTES expression in the culture of nasal polyp fibroblasts. MATERIAL AND METHODS: NP fibroblast cell cultures derived from 16 patients with NP were first stimulated with bacterial LPS and than incubated in increasing concentrations (from 10(-7)M to 10(-4)M) of calcitriol, tacalcitol or budesonide and in combination with one of VD derivate with budesonide in 1:1, 1:3 and 3:1 ratios. Quantitative analysis of RANTES level was conducted in culture supernatants using an ELISA method. RESULTS: The highest calcitriol concentration (10(-4)M) as well as tacalcitol at 10(-5)M and 10(-4)M reduced RANTES production significantly compared to the control (201.1pg/ml, 338.7pg/ml, 211.3pg/ml v 571.78pg/ml; p<0.05). Budesonide and calcitriol administered in 1:3 ratio and budesonide and tacalcitol in 1:1 and 1:3 reduced RANTES concentration significantly better than each of the drug used in monotherapy (p<0.05). Budesonide and tacalcitol in 1:1 and 1:3 ratios suppressed RANTES production to the lowest level (171.8±97.6pg/ml and 178.7±105.22pg/ml, respectively). CONCLUSION: Active VD compounds via downregulation of RANTES production exert a potential role as a complementary element in the therapy of chronic rhinosinusitis with NP. Compounds consisting of budesonide and VD derivate have an advantage over both drugs used in monotherapy.

Vitamin D derivatives: calcitriol and tacalcitol inhibits interleukin-6 and interleukin-8 expression in human nasal polyp fibroblast cultures.

PURPOSE: Biologically active vitamin D3 (VD) derivatives possess modulatory activities on immunological and inflammatory responses which can be reflected by altered levels of pro-inflammatory chemokines. Nasal polyposis (NP), defined as a chronic inflammatory process of upper respiratory system, could be influenced by VD derivatives. The purpose of this study was to investigate the influence of 1alpha,25-dihydroxyvitamin D3 (calcitriol) and 1alpha,24(R)-dihydroxyvitamin D3 (tacalcitol) on the secretion of IL-6 and IL-8 by fibroblasts derived from NP. MATERIAL AND METHODS: The study involved 12 fibroblast cultures derived from NP samples obtained from surgically treated patients. Measurements were performed on the polyp cells after the 6-9 passages. Culture stimulation involved treatment with tacalcitol and calcitriol at a defined strength (from 10(-7)M to 10(-4)M). IL-6 and IL-8 concentrations were estimated with ELISA. RESULTS: Treatment with calcitriol or tacalcitol inhibits the synthesis of both IL-6 and IL-8 compared to the control group. The dose dependence of this effect has been confirmed. VD derivatives influence was marked at higher concentrations. Significant interleukin decrease was observed at 10(-5) and 10(-4) for calcitriol and 10-4 in the case of tacalcitol. CONCLUSIONS: The present study demonstrates that calcitriol and tacalcitol are capable of affecting pro-inflammatory cytokine (IL-6 and IL-8) levels in NP cultures. Our data imply a potential therapeutical application of topical VD derivates in NP and warrant further investigation.

[The quantitative analysis of E6 HPV and P53 genes expression by QPCR (TaqMan) in the assessment of surgical treatment range in vulvar carcinoma induced by HTV infection]. / Ilosciowa analiza ekspresji genów E6 HPV 18 i P53 metoda QPCR (TaqMan) w ocenie marginesu operacyjnego w raku sromu indukowanym infekcja HPV.

OBJECTIVES: Human Papilloma Virus (HPV) infections especially 16 and 18 are risk factors for squamous cell vulvar cancer. E6 protein of HPV joins the tumor suppressor protein P53 and promotes its degradation. This is one of the possible mechanisms of viral oncogenes action. DESIGN: In this study the quantitative analysis of mRNA copies E6 HPV 18 and mRNA P53 expression in vulvar cancer tissue was performed. The expression of analysed genes was applied in the assessment of surgical treatment range. MATERIALS AND METHODS: The specimens from a 26 year old woman with vulvar squamous cell cancer stage II FIGO treated surgically modo Way were analysed. The number of DNA and mRNA copies E6 HPV and P53 were examined basing on Q-PCR standard curves for beta-actine by use of Perkin Elmer-kit and the sequence detector ABI PRISM 7700-Taq Man application. RESULTS: The overexpression of mRNA E6 HPV and P53 in analysed specimens was found. The highest number of mRNA copies in cancer tissue was ascertained. In lichen sclerosus and lymphonoduli tissue lower number of analysed copies was found. CONCLUSION: The quantitative analysis of E6 HPV and P53 genes expression can be useful in the assessment of surgical treatment range in vulvar cancer and can also be used as a prognostic marker.

[Expression of VEGF, KDR, p53, E6 HPV16 and HPV18 in vulvar and cervix cancer]. / Ekspresja genów angiogenezy--VEGF i jego receptora KDR, genu supresorowego P53 oraz E6 HPV16 i HPV18 w raku sromu i szyjki macicy.

The aim of the study was a comparison of expression of angiogenesis genes: vascular endothelial growth factor (VEGF), KDR, suppressor gene p53, E6-HPV16 and HPV18, in tissue samples of normal, dystrophic, lymph nodes and malignant cancers of vulva and uterine cervix. The results demonstrate that molecular diagnostics of cancers using gene expression profiling indicates the definitive difference in expression profiles of aforementioned genes in tissues of the same malignancy.

[Profiles of expression of genes coding kininogen and kinin receptors as a marker of tissue pathology in cervical cancer coexisting with HPV infection]. / Ekspresja genów receptorów dla kinin i kininogenu w raku szyjki macicy wspólistniejacym z infekcja HPV.

Kinins are peptides involved in inflammatory processes, vascular permeability, proliferation and mitogenesis of tumor cells. The majority of kinins actions are mediated through an interaction with cell surface bradykinin receptors BR1 and BR2. Kinins precursor is kininogen (kng). The changes in proteins are initiated by changes in the expression of genes coding these proteins, thus can be a valuable diagnostic markers of malignant processes including cervical carcinoma. The paper presents an analysis of kininogen-kinins receptor genes expression in women treated surgically because of a carcinoma of uterine cervix. Among the studied women in 5 cases previously brachyHDR therapy was applied. In all studied cases HPV 18 infection and in 2 cases a co-infection HPV 16/18 by use of Consensus Primers MY09, MY 11 and type specific primers for HPV 16, 18 was ascertained. In RNA extracts the number of the mRNA copies for kiningen, BR1 and BR2 was assessed using QRT-PCR Taq Man. The higher expression of BR1 than BR2 was marked in the tissue with cancer cells. In the patients after brachytherapy higher expression of BR2 than BR1 mRNA was found. The higher BR1 expression was also shown in iliac lymph nodes in patients with active neoplastic process, opposite to the patients after brachytherapy in whom higher BR2 expression was ascertained. The lack of expression of kng mRNA was found only in 3 specimens. The high expression of kinin receptors especially BR1 in infiltrating carcinoma margin can be a marker of pathology intensity: proliferative potential of neoplasms cells or chronic inflammatory state in the presence of invasive carcinoma.

Differences between Desulfovibrio desulfuricans intestinal strains with respect to their ability of sulphasalazine biotransformation.

In presented study, differences among six intestinal Desulfovibrio desulfuricans strains were investigated regarding their ability of sulphasalazine (SAS) biotransformation. Bacterial strains were incubated (24 or 48 h) in liquid medium supplemented with SAS (12.2-150 microM). It has been demonstrated that investigated D. desulfuricans strains converted SAS with various rates, depending on the strain used and on time of drug contact with bacterial cells. Significantly different (50% coefficient of variability) were the maximum rates of SAS biotransformation. It was demonstrated that these rates were strongly dependent on the drug concentration. Increasing SAS concentrations up to about 120 microM caused increase in the drug transformation rate. At higher SAS concentrations this rate remained at the same level or was inconsiderable lower. The strains exposed to SAS for 24 hs transformed this drug with the rate about twice as compared with that observed after 48 h exposition.