Journal of Molecular Biomarkers & Diagnosis

We hypothesized that leucocytes may be a significant source of physiologically active gases. Furthermore, it is hoped that alterations by disease of these volatile organic compound (VOC) profiles can ultimately be used as noninvasive biomarkers. We recently demonstrated that transformed promyelocitic cells produce distinct gases. (J Transl Med. 2009 7:31). The current study extends this work to neutrophils and peripheral blood mononuclear cells (PBMCs). Cells were isolated from the peripheral blood of healthy donors (n=10, 18-65 yrs old), resuspended in RPMI, and incubated in bioreactors for 24 hrs. The headspace was analyzed using gas chromatography. Acetaldehyde was elevated from neutrophils(median (min, max); 197 (34, 577) ppbv) compared with media (88 (40,116) ppbv, p=0.014). In the presence of alanine, neutrophils emitted more acetaldehyde (>1.5 fold compared to basal level). In contrast, acetaldehyde from PBMCs was 26 (8, 89) ppbv, significantly below media (p=0.004). Adding alanine did not affect acetaldehyde emissions from PBMCs. Also, hexanaldehyde appeared metabolized by neutrophils and PBMCs. This study demonstrates 1) that human primary immune cells produce measurable VOCs in vitro, and 2) the ability to detect basal levels of acetaldehyde from unperturbed cultured neutrophils. Moreover, the data suggest that different leukocyte subtypes have different VOC profiles.

Epithelial ovarian cancer is morphologically heterogeneous being classified as serous, endometrioid, clear cell, or mucinous. Molecular genetic analysis has suggested a role for tumor suppressor genes located at chromosome 10q in epithelial ovarian cancer pathogenesis. Our objective is to evaluate the expression of ANXA7, a novel tumor suppressor gene located on 10q21, in these epithelial ovarian cancer subtypes, and to investigate its correlation with patient survival. ANXA7 is ubiquitously expressed in small amounts in nearly every normal cell and the Anxa7 (+/-) knockout mouse has a cancer prone phenotype. Altered ANXA7 protein levels are associated with prognostically challenging aggressive forms of prostate and breast cancer. So far, information is not available regarding the association of ANXA7 expression in ovarian cancer and patient survival. Therefore, we used human tumor tissue microarray (TMA) technology in order to evaluate the ANXA7 immunoreactivity as possible diagnostic and/or prognostic marker of ovarian cancer by immunoperoxidase assay using ANXA7 monoclonal antibody. Using a 129 case diagnostic human tumor tissue microarray, we report that the expression of ANXA7 is significantly reduced and is associated with disease progression. Furthermore, using a separate 301 case retrospective prognostic tumor tissue microarray, we find that loss of ANXA7 expression is also significantly associated with poor over-all patient survival. We conclude that ANXA7 may be a new prognostic marker or a target for improving the treatment efficiency of patients with ovarian cancers.

Objectives: Previously, we reported that neutrophil gelatinase-associated lipocalin (NGAL) is present in epithelial ovarian cancer cells, but not mesenchymal cancer cells. Furthermore, we reported that epithelial growth factor (EGF) induced epithelio-mesenchymal transition (EMT) in epithelial ovarian cancer cells and concomitantly suppressed NGAL expression. We hypothesised that inhibition of NGAL expression induces EMT in these cells.

Methods: NGAL knockdown was achieved using siRNA in ovarian cancer cell line OVCA 429. NGAL siRNAtreated cells [n=3] were assayed for EMT markers such as migration and invasion assays, and E-cadherin, N-cadherin, Snail, ?-catenin and vimentin expression by Western blot.

Results: There were no significant differences between NGAL siRNA-treated cells and controls for the protein markers. There was an increase in cell migration for NGAL siRNA-treated cells compared to control (96 h post transfection), however there was no change in cell invasiveness between treatments.

Conclusions: We conclude that direct NGAL inhibition by siRNA does not induce a mesenchymal transition of ovarian cancer cells.

CD40 plays a substantial role in inflammation and has been linked to pathogenic processes of chronic inflammatory diseases such as asthma as well as chronic obstructive pulmonary disease (COPD). Aim: The study was to investigate the association of CD40 gene (-1C/T) single nucleotide polymorphism (SNP) with the susceptibility to asthma and COPD in the Egyptian population, and its functional effect on the expression of CD40. Methods: We analyzed -1C/T SNP of the CD40 gene in 40 patients with COPD, 50 patients with asthma, and 60 normal subjects using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). CD40 expression was measured using flow cytometry. Immunoglobulin E was also determined in asthmatics. Results: The CT genotype was prevailing in the COPD patients and control group, while in the asthmatics it was the CC. Independent of the smoking status; being CC homozygous still conferred a 4-fold increase in the risk of asthma and a 2.5-fold increase in the risk of COPD. Carrying T allele showed a significantly lower risk for asthma. Furthermore, in both asthma and COPD, the least expression of CD40 protein was found with the TT genotype, which seems to have a protective effect. Despite the significant upregulation of total serum IgE in asthmatics it was not significantly associated with CD40 genotyping or the protein expression. Conclusion: Our study demonstrated that CD40 ? 1C/T polymorphism significantly contribute to the susceptibility to asthma and COPD in the Egyptian population. Reduced CD40 expression with the TT genotypes might imply that the ? 1C/T polymorphism is linked to inflammation in addition to the initiation and development of both. The genetic predisposition to certain pathways may further help to define the development of either asthma or COPD. This may lead to stratification of patients by their genetic make-up and open new therapeutic prospects.