Disease Markers - Volume 29, issue 5

Authors: Kruszewski, Wiesław Janusz | Rzepko, Robert | Ciesielski, Maciej | Szefel, Jarosław | Zieliński, Jacek | Szajewski, Mariusz | Jasiński, Wojciech | Kawecki, Krzysztof | Wojtacki, Janusz

Article Type: Research Article

Abstract: Estimation of HER2 membranous expression is routinely used in breast and gastric cancers, as both a prognostic and a predictive factor. To date there is no evidence for similar application of HER2 expression in colorectal cancer (CRC) cells. In CRC, HER2 is sometimes overexpressed in the cell membrane and very often in the cytoplasm. This study was conducted to determine possible correlations between both membranous and cytoplasmatic expression of HER2 in CRC cells and the outcome …of the disease. The prognostic significance of combined staining intensity in the cell membrane and cytoplasm in the entire CRC cell was also investigated. HER2 expression in resectable colorectal adenocarcinoma cells was evaluated by immunohistochemistry in specimens taken from 202 patients. The percentage of cancer cells with membranous or cytoplasmatic reactions and the staining intensity of the reaction in the whole cell were recorded. A membranous reaction was present in 27% of cases, and cytoplasmatic reaction in 66% of cases. The total staining intensity in the entire cell was evaluated as moderate (2+) in 32% of cases and strong (3+) staining in 15%. There was no correlation found between either membranous or cytoplasmatic HER2 expression and survival. Furthermore combined staining intensity did not provide any prognostic information. We conclude that HER2 expression in CRC does not correlate with prognosis. Show more

Keywords: HER2, colorectal cancer, prognosis

DOI: 10.3233/DMA-2010-0742

Citation: Disease Markers, vol. 29, no. 5, pp. 207-212, 2010

Authors: Sandhya, Narasimhan | Gokulakrishnan, Kuppan | Ravikumar, Radhakrishnan | Mohan, Viswanathan | Balasubramanyam, Muthuswamy

Article Type: Research Article

Abstract: Objective: The aim of this study is to measure the extent of oxidative stress and to see whether it has any correlation to changes in adiponectin levels in NAFLD subjects with and without Type 2 diabetes. Methods: Subjects recruited from the Chennai Urban Rural Epidemiology Study comprise of 1: Normal Glucose Tolerance (NGT) subjects without NAFLD; 2: NGT with NAFLD; 3: Type 2 Diabetic patients [T2DM] without NAFLD and 4: T2DM with NAFLD. Thiobarbituric acid reactive substances (TBARS), …protein carbonyl (PCO), glutathione and adiponectin levels were measured by standard methods. Ultrasound of the liver was used to diagnose NAFLD. Results: T2DM subjects with NAFLD had significantly (p< 0.001) higher levels of thiobarbituric acid reactive substances (TBARS) and protein carbonyls (PCO) but lower (p< 0.001) GSH/GSSG ratio and adiponectin levels compared to other three groups. The association of hypoadiponectinemia with NAFLD/Type 2 diabetes was significant even after adjusting for age, gender and BMI, but lost when adjusted for parameters of oxidative stress. While palmitate significantly reduced GSH/GSSG ratio in hepatocytes, addition of exogenous recombinant adiponectin restored the GSH/GSSG ratio comparable to those of untreated cells. Conclusion: There exists an association of hypoglutathionemia and hypoadiponectinemia in subjects with NAFLD and/or T2DM. In addition to the known beneficial effects, out study also exposes the antioxidant nature of adiponectin. Show more

Keywords: Type 2 diabetes, NAFLD, glutathione, adiponectin, oxidative stress

DOI: 10.3233/DMA-2010-0743

Citation: Disease Markers, vol. 29, no. 5, pp. 213-221, 2010

Authors: Joshi, Gunjan | Pradhan, Sunil | Mittal, Balraj

Article Type: Research Article

Abstract: We aimed to find out if the serotonin receptor (HT102T>C) and serotonin transporter (STin 2) polymorphisms play any role in genetic susceptibility of migraine. For the study, 217 migraine patients and 217 healthy controls (HC) were recruited and genotyping was carried out using the Polymerase Chain Reaction and Restriction Fragment Length polymorphism (PCR-RFLP) method. All results were Bonferroni corrected. We could not find any significant differences in the genotype or allele frequencies …in case of HT 102 T>C polymorphism between migraine patients and healthy controls (P value=0.224). No significant association was seen at allele and carrier levels. Sub-grouping the patients on the basis of gender or on basis of migraine type i.e. with or without aura also did not show any association. Similarly, no difference in genotype (P value=0.236), allele (P value=0.550) or carrier frequency (P value=0.771) in STin 2 VNTR polymorphism was observed between migraine patients. However, HT 102 TC genotype was observed to interact significantly with the STin 2.10/10 genotype in enhancing risk of migraine, both with and without aura. In conclusion, the HT102 T>C receptor and the STin 2 VNTR transporter polymorphisms, did not individually confer any significant risk of migraine or its clinical subtypes but the two polymorphisms appear to synergistically influence susceptibility to migraine. Serotonin transporter (STin2 VNTR) and receptor (HT 102T>C) polymorphisms; Migraine with aura (MA); Migraine without aura (MO); Genetic susceptibility Show more

DOI: 10.3233/DMA-2010-0752

Citation: Disease Markers, vol. 29, no. 5, pp. 223-229, 2010

Authors: Liang, Seng | Singh, Manjit | Dharmaraj, Saravanan | Gam, Lay-Harn

Article Type: Research Article

Abstract: Breast cancer is a leading cause of mortality in women. In Malaysia, it is the most common cancer to affect women. The most common form of breast cancer is infiltrating ductal carcinoma (IDC). A proteomic approach was undertaken to identify protein profile changes between cancerous and normal breast tissues from 18 patients. Two protein extracts; aqueous soluble and membrane associated protein extracts were studied. Thirty four differentially expressed proteins were identified. The intensities of the proteins …were used as variables in PCA and reduced data of six principal components (PC) were subjected to LDA in order to evaluate the potential of these proteins as collective biomarkers for breast cancer. The protein intensities of SEC13-like 1 (isoform b) and calreticulin contributed the most to the first PC while the protein intensities of fibrinogen beta chain precursor and ATP synthase D chain contributed the most to the second PC. Transthyretin precursor and apolipoprotein A-1 precursor contributed the most to the third PC. The results of LDA indicated good classification of samples into normal and cancerous types when the first 6 PCs were used as the variables. The percentage of correct classification was 91.7% for the originally grouped tissue samples and 88.9% for cross-validated samples. Show more

Keywords: Breast cancer, proteomics, biomarkers, PCA, LDA

DOI: 10.3233/DMA-2010-0753

Citation: Disease Markers, vol. 29, no. 5, pp. 231-242, 2010

Authors: Wee, Line | Vefring, Hege | Jonsson, Grete | Jugessur, Astanand | Lie, Rolv Terje

Article Type: Research Article

Abstract: Preeclampsia is a serious disorder affecting nearly 3% of all in the Western world. It is associated with hypertension and proteinuria, and several lines of evidence suggest that the renin-angiotensin system (RAS) may be involved in the development of hypertension at different stages of a preeclamptic pregnancy. In this study, we developed rapid genotyping assays on the LightCycler® instrument to allow the detection of genetic variants in the renin gene (REN) that may predispose …to preeclampsia. The method is based on real-time PCR and allele-specific hybridization probes, followed by fluorescent melting curve analysis to expose a change in melting temperature (T_{m} ). Ninety-two mother-father-child triads (n=276) from preeclamptic pregnancies were genotyped for three haplotype-tagging single nucleotide polymorphisms (htSNPs) in REN. All three htSNPs (rs5705, rs1464816 and rs3795575) were successfully genotyped. Furthermore, two unexpected nucleotide substitutions (rs11571084 and rs61757041) were identified within the selected hybridization probe area of rs1464816 and rs3795575 due to aberrant melting peaks. In conclusion, genotyping on the LightCycler® instrument proved to be rapid and highly reproducible. The ability to uncover additional nucleotide substitutions is particularly important in that it allows the identification of potentially etiological variants that might otherwise be overlooked by other genotyping methods. Show more

Keywords: Preeclampsia, renin, LightCycler, real-time PCR

DOI: 10.3233/DMA-2010-0754

Citation: Disease Markers, vol. 29, no. 5, pp. 243-249, 2010

Authors: Guo, Hong-Qiang | Huang, Guo-Liang | Guo, Cheng-Cheng | Pu, Xing-Xiang | Lin, Tong-Yu

Article Type: Research Article

Abstract: Extra-nodal natural killer T-Cell (NK/T-cell) lymphoma is a progressive cancer with poor prognosis due to the lack of disease specific treatment. To develop specific therapeutic strategies, it is essential to identify tumor markers. Recent studies show that circulating microRNA (miRNA) may serve as diagnostic and/or prognostic markers for some diseases. To explore miRNAs as potential diagnostic and/or prognostic markers of NK/T-cell lymphoma, in our study, we compared circulating miR-221 levels in 79 patients and 37 …normal subjects by real-time PCR amplification directly from plasma samples, and correlated patient's miR-221 levels with their clinic features and treatment outcomes. We observed a significant difference between the patient and control groups (p=0.038), and a correlation of plasma miR-221 level in patient with sex, as well as a reverse correlation with performance status and the overall survival after treatment. Univariate and multivariate analyses further revealed that plasma miR-221 level, age, B symptoms, LDH level and complete response after primary treatment all present prognostic values when judged by overall survival (OS). Together, our results show that it is feasible to perform direct amplification of plasma miRNAs without total RNA extraction, and plasma miR-221 may be a diagnostic and prognostic marker for NK/T-cell lymphoma. Show more

Keywords: microRNA, NK/T-cell lymphoma, prognosis, quantitative real-time PCR

DOI: 10.3233/DMA-2010-0755

Citation: Disease Markers, vol. 29, no. 5, pp. 251-258, 2010

Authors: Galgani, Andrea | Valdes, AnaMaria | Erlich, Henry A. | Mano, Calvin | Cheng, Suzanne | Petrone, Antonio | Sentinelli, Federica | Berni, Andrea | Baroni, Marco G. | Buzzetti, Raffaella

Article Type: Research Article

Abstract: Several studies suggest that the peroxisome proliferator-activated receptor gamma (PPARγ) is involved in atherogenesis. The Pro12Ala polymorphism in the gene encoding PPARγ (PPARγ2 gene) influences the risk for type 2 diabetes. Two population-based studies have shown that the Ala allele is associated with reduced carotid intimal-medial thickness (IMT). However, studies focusing on acute clinical events have yielded conflicting results. Our aim was to evaluate the role of the Pro12Ala PPARγ2 polymorphism on …the risk of coronary artery disease (CAD) in an Italian population with a case-controlled genetic association study in which 478 CAD patients and 218 controls were genotyped for the Pro12Ala polymorphism. CAD was diagnosed by angiography. We found that homozygotes for the Ala12 allele had a significantly reduced risk of CAD after adjusting for diabetes, sex, age, body mass index (BMI), smoking, lipids and hypertension (OR =0.007; 95% C.I. = 0.00–0.32 p< 0.011). In this case-control study, homozygosity for the Ala allele at codon 12 of the PPARγ2 gene resulted in reduced risk of CAD. This is consistent with reports from previous studies focusing on atherosclerosis and myocardial infarction. Show more

Keywords: Coronary artery disease (CAD), Peroxisome Proliferator-Activated Receptor Gamma 2 (PPARγ2), Myocardial infarction (MI), Pro12Ala polymorphism

DOI: 10.3233/DMA-2010-0756

Citation: Disease Markers, vol. 29, no. 5, pp. 259-264, 2010

Authors: Verschuren, J.J.W. | Sampietro, M.L. | Pons, D. | Trompet, S. | Ewing, M.M. | Quax, P.H.A. | de Knijff, P. | Zwinderman, A.H. | de Winter, R.J. | Tio, R.A. | de Maat, M.P. | Doevendans, P.A.F.M. | Jukema, J.W.

Article Type: Research Article

Abstract: Objective: Mixed results have been reported of matrix metalloproteinases (MMP) and their association with restenosis after percutaneous coronary intervention (PCI). The current study examines whether multiple single nucleotide polymorphisms (SNPs), covering the full genomic region of MMP2 and MMP3, were associated with restenosis in the GENDER study population. Methods and results: The GENetic DEterminants of Restenosis (GENDER) study enrolled 3104 consecutive patients after successful PCI. The primary endpoint was clinical …restenosis, defined as target vessel revascularization (TVR), occurring in 9.8% of the patients. From the Hapmap database, 19 polymorphisms of MMP2 and 11 of MMP3 were selected. Furthermore, in a subpopulation, a genome-wide association analysis (GWA) was performed. No significant association was found with any of the investigated SNPs, including the previously reported 5A/6A polymorphism (rs3025058), with regard to TVR using single SNP analysis or haplotype analysis. Conclusion: We found no significant association of MMP2 or MMP3 with TVR with this SNP-broad gene approach. Although we did not test all the known polymorphisms of these genes, using tagging analyses we examined those SNPs covering all known haplotypes of MMP2 and MMP3 to conclude that these genes do not correlate with a genetic risk of coronary restenosis after successful PCI. Show more

Keywords: Matrix metalloproteinases, percutaneous coronary intervention, restenosis, single nucleotide polymorphisms

DOI: 10.3233/DMA-2010-0757

Citation: Disease Markers, vol. 29, no. 5, pp. 265-273, 2010