VKORC1 gene (vitamin K epoxide reductase) polymorphisms are associated with cardiovascular disease in chronic kidney disease patients on hemodialysis.

Vitamin K is necessary for the carboxylation of clotting factors and matrix Gla protein (MGP). Vitamin K epoxide reductase (VKOR) is the enzyme responsible for recirculation of Vitamin K increasing its tissue availability. Polymorphisms of VKOR may alter the function of MGP, thereby influencing vascular calcification. We conducted this study to investigate the relationship of VKORC1 gene single nucleotide polymorphisms (SNP's) to vascular calcification and clinically overt cardiovascular disease in chronic kidney disease (CKD) patients on hemodialysis (HD). The study included 54 CKD patients on HD. We excluded those with diabetes or on anticoagulant therapy. Vascular calcifications were measured using computerized tomography scans and roentgenograms. Prevalent clinically overt cardiovascular disease was reported based on the evidence of documented preexisting major cardiovascular events. Genotype detection for the gene VKORC1 C1173T and G-1639A polymorphisms was carried out by polymerase chain reaction. We found a significant association between C1173T polymorphisms and vascular calcification (odds ratio [OR] = 43, P = 0.001). The mutant T allele was also linked with higher odds of vascular calcification (OR = 8.880, 95% confidence interval [CI] = 3.1-25.4, P = 0.001) and clinically overt cardiovascular disease (OR = 4.7, 95% CI = 1.5-14.7, P = 0.005). VKORC1 G-1639A polymorphisms were not associated with vascular calcification and had lower prevalence of clinically overt cardiovascular disease (OR = 0.07, 95% CI = 0.01-0.4, P = 0.001). In patients with CKD on HD, we found that VKORC1 gene polymorphisms did have an association with prevalent cardiovascular calcification and clinically overt cardiovascular disease, C1173T polymorphisms with higher risk for disease, and G-1639A with lower risk.

Increased Free Circulating DNA Integrity Index as a Serum Biomarker in Patients with Colorectal Carcinoma.

BACKGROUND: Cell-free DNA circulating in blood is a candidate biomarker for malignant tumors. Unlike uniformly truncated DNA released from apoptotic non diseased cells, DNA released from necrotic cancer cells varies in size. OBJECTIVES: To measure the DNA integrity index in serum and the absolute DNA concentration to assess their clinical utility as potential serum biomarkers for colorectal carcinoma (CRC) compared to CEA and CA19-9. MATERIALS AND METHODS: Fifty patients with CRC, 10 with benign colonic polyps and 20 healthy sex and age matched volunteers, were investigated by real time PCR of ALU repeats (ALU q-PCR) using two sets of primers (115 and 247 bp) amplifying different lengths of DNA fragments. The DNA integrity index was calculated as the ratio of q-PCR results of ALU 247/ ALU 115bp. RESULTS: Serum DNA integrity was statistically significantly higher in CRC patients compared to the benign and control groups (p<0.001). ROC curves for differentiating CRC patients from normal controls and benign groups had areas under curves of 0.90 and 0.85 respectively. CONCLUSIONS: The DNA integrity index is superior to the absolute DNA concentration as a potential serum biomarker for screening and diagnosis of CRC. It may also serve as an indicator for monitoring the progression of CRC patients. Combining CEA and CA19-9 with either of the genetic markers studied is better than either of them alone.

Evaluation of annexin A2 and as potential biomarkers for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) ranks as the fifth most common malignancy worldwide. Early detection of HCC is difficult due to the lack of reliable markers. We aimed to assess the diagnostic role of annexin A2 (ANXA2) and follistatin as serum markers for HCC patients. This study included 50 patients with confirmed diagnosis of HCC, 30 patients with chronic liver disease, and 20 normal persons. Subjects performed thorough assessment and laboratory investigations. Serum levels of alpha fetoprotein (AFP), annexin A2, and follistatin were measured using ELISA technique. Annexin A2 significantly increased in the sera of HCC patients (median, 69.6 ng/ml) compared to chronic liver disease patients (median, 16.8 ng/ml) and control group (median, 9.5 ng/ml) (p < 0.001). Follistatin was higher in sera of HCC patients (median, 24.4 ng/ml) compared to the control group (median, 4.2 ng/ml) (p = 0.002) while no such significant difference was achieved between HCC and chronic liver disease patients. At a cutoff level 29.3 ng/ml, area under the receiver-operating characteristic curve for ANXA2 was 0.910 (95 % confidence interval (CI) 0.84-0.97). For follistatin, it was 0.631 (95 % confidence interval 0.52-0.74) at cutoff level 15.7 ng/ml. Combining both annexin A2 and AFP increased the diagnostic efficiency (98 % specificity, LR + 41 and 97.6 % PPV). Follistatin combined with AFP provided 92 % specificity while lower sensitivity (50 %) was observed. Serum ANXA2 is a promising biomarker for HCC, certainly when measured with AFP. Follistatin could not differentiate between HCC and chronic liver disease, but its combination with AFP improved the specificity for HCC diagnosis.

Circulating miRNA-122, miRNA-199a, and miRNA-16 as Biomarkers for Early Detection of Hepatocellular Carcinoma in Egyptian Patients with Chronic Hepatitis C Virus Infection.

BACKGROUND: Hepatocellular carcinoma (HCC) is the sixth most common cancer in the world. Having a very poor prognosis, it currently ranks as the third most common cause of cancer-related deaths. MiRNAs are a set of small, single-stranded, non-coding RNA molecules that negatively regulate gene expression at the post-transcriptional level. Several miRNAs were found to be frequently deregulated in HCC. OBJECTIVE: To investigate whether miRNA-122, miRNA-199a, and miRNA-16 are altered in sera of hepatitis C virus (HCV)-induced HCC patients compared with chronic HCV patients without HCC, and to assess their diagnostic value to differentiate between HCC and chronic HCV in order to develop a non-invasive diagnostic and prognostic tool for HCC. METHODS: We analysed the expression of mature miRNA-122, miRNA-199a, and miRNA-16 in serum by a singleplex TaqMan two-step stem loop quantitative real-time reverse-transcription PCR (qRT-PCR) in 40 newly diagnosed HCC patients and 40 chronic HCV liver cirrhosis patients, as well as 20 apparently healthy individuals as a control group, using RNU48 as a normalisation control. RESULTS: Serum miR-16 was significantly lower in HCC than in HCV patients (P = 0.033). The serum level of miR-199a in chronic HCV patients was significantly lower than in healthy controls (P = 0.001). Receiver operating curve (ROC) analysis for serum miRNA-16 for discriminating HCC from HCV patients showed that at the cut-off value of 0.904, the sensitivity and specificity for this marker were 57.5 and 70 %, respectively. The combination of serum miR-16 with serum alpha fetoprotein (AFP) resulted in improved sensitivity to 85% and increased diagnostic accuracy to 87.5 %. Serum miR-199a and miR-16 were significantly associated with several parameters of HCC such as tumour size and number. CONCLUSION: The combination of serum miR-16 and serum AFP is a significant improvement on the current best practice of serum AFP for HCC in HCVpositive patients. Serum miR-199a and miR-16 could be used as potential indicators of the progress of HCC.