Quantification of cell-free circulating mitochondrial DNA copy number variation in hepatocellular carcinoma.

OBJECTIVE: Hepatocellular carcinoma is the most common primary malignant liver tumor. Mitochondrial DNA copy number has been shown to be associated with various malignancies. However, there has not been any study on the absolute quantification of mtDNA copy number in hepatocellular carcinoma. The aim of this study was to develop a new method for absolute quantification of mtDNA copy number and to relatively quantify the variations in the mtDNA copy number in hepatocellular carcinoma patients in comparison with healthy individuals. METHODS: Venous blood samples were collected from both hepatocellular carcinoma patients (34) and healthy individuals (34). Circulating cell-free DNAs were isolated and the relative quantification of mtDNA copy number variation was determined using quantitative polymerase chain reaction and digital polymerase chain reaction. RESULTS: It was found that the relative mtDNA copy number was significantly decreased in hepatocellular carcinoma patients in comparison with the control group (p<0.05). The median (range) and average of relative mtDNA/ß-actin gene of the patients were determined as 42.8 cp/µL (11.1-88.5) and 45.1 cp/µL, respectively, while the median (range) and average relative mtDNA/ß-actin gene of the control group were determined as 102.8 cp/µL (55.1-291.8) and 138.7 cp/µL, respectively (p<0.05). When quantitative polymerase chain reaction and digital polymerase chain reaction were compared, mtDNA/ß-actin gene copy number ratio of digital polymerase chain reaction results was found to be 1.76-fold more than that of quantitative polymerase chain reaction results. CONCLUSION: Circulating mtDNA copy number was decreased in hepatocellular carcinoma patients in comparison with healthy individuals, and we suggest that it can be used as a noninvasive biomarker for hepatocellular carcinoma diagnosis in the future.

Characteristics of patients with hepatocellular carcinoma: A multicenter study.

Background and Aim: The aim of the present study was to examine the etiology of hepatocellular carcinoma (HCC) by underlying cause and determine the characteristics and clinical features of patients with HCC. Materials and Methods: The study comprised 1802 HCC patients diagnosed and followed up by Liver Diseases Outpatient Clinics in 14 tertiary centers in Turkey between 2001 and 2020. Results: The mean age was 62.3±10.7 years, and 78% of them were males. Of the patients, 82% had cirrhosis. Hepatitis B virus (HBV) infection was the most common etiology (54%), followed by hepatitis C virus (HCV) infection (19%) and nonalcoholic fatty liver disease (NAFLD) (10%). Of the patients, 56% had a single lesion. Macrovascular invasion and extrahepatic spread were present in 15% and 12% of the patients, respectively. The median serum alpha-fetoprotein level was 25.4 ng/mL. In total, 39% of the patients fulfilled the Milan Criteria. When we compared the characteristics of patients diagnosed before and after January 2016, the proportion of NAFLD-related HCC cases increased after 2016, from 6.6% to 13.4%. Conclusion: Chronic HBV and HCV infections remain the main causes of HCC in Turkey. The importance of NAFLD as a cause of HCC is increasing.

Quantification of cell-free circulating mitochondrial DNA copy number variation in hepatocellular carcinoma

SUMMARY OBJECTIVE: Hepatocellular carcinoma is the most common primary malignant liver tumor. Mitochondrial DNA copy number has been shown to be associated with various malignancies. However, there has not been any study on the absolute quantification of mtDNA copy number in hepatocellular carcinoma. The aim of this study was to develop a new method for absolute quantification of mtDNA copy number and to relatively quantify the variations in the mtDNA copy number in hepatocellular carcinoma patients in comparison with healthy individuals. METHODS: Venous blood samples were collected from both hepatocellular carcinoma patients (34) and healthy individuals (34). Circulating cell-free DNAs were isolated and the relative quantification of mtDNA copy number variation was determined using quantitative polymerase chain reaction and digital polymerase chain reaction. RESULTS: It was found that the relative mtDNA copy number was significantly decreased in hepatocellular carcinoma patients in comparison with the control group (p<0.05). The median (range) and average of relative mtDNA/β-actin gene of the patients were determined as 42.8 cp/μL (11.1-88.5) and 45.1 cp/μL, respectively, while the median (range) and average relative mtDNA/β-actin gene of the control group were determined as 102.8 cp/μL (55.1-291.8) and 138.7 cp/μL, respectively (p<0.05). When quantitative polymerase chain reaction and digital polymerase chain reaction were compared, mtDNA/β-actin gene copy number ratio of digital polymerase chain reaction results was found to be 1.76-fold more than that of quantitative polymerase chain reaction results. CONCLUSION: Circulating mtDNA copy number was decreased in hepatocellular carcinoma patients in comparison with healthy individuals, and we suggest that it can be used as a noninvasive biomarker for hepatocellular carcinoma diagnosis in the future.

Validation of the PAGE-B score to predict hepatocellular carcinoma risk in caucasian chronic hepatitis B patients on treatment.

BACKGROUND: Several risk scores have been developed to predict hepatocellular carcinoma (HCC) risk in chronic hepatitis B (CHB) patients. The majority of risk scores are based on pretreatment variables that are no longer considered risk factors for HCC development due to the suppression of hepatitis B virus replication early in the course of potent antiviral treatment in most patients. The PAGE-B score, which is based on platelet levels, age and sex, has been shown to accurately predict HCC risk in CHB patients on antiviral treatment in various populations. AIM: We aimed to evaluate the PAGE-B score in predicting HCC risk in Turkish CHB patients on antiviral treatment. METHODS: In this study, we recruited 742 CHB patients who had been treated with tenofovir disoproxil fumarate or entecavir for ≥ 1 year. Risk groups were determined according to the PAGE-B scores as follows: ≤ 9, low; 10-17, moderate and ≥ 18, high. The cumulative HCC incidences in each risk group were computed using Kaplan-Meier analysis and were compared using the log-rank test. The accuracy of the PAGE-B score in predicting HCC risk was evaluated using a time-dependent area under the receiver operating characteristic (AUROC) curve at all study time points. Univariate and multivariate logistic regression analyses were used to assess the risk factors for HCC development. RESULTS: The mean follow-up time was 54.7 ± 1.2 mo. HCC was diagnosed in 26 patients (3.5%). The cumulative HCC incidences at 1, 3, 5 and 10 years were 0%, 0%, 0% and 0.4% in the PAGE-B low-risk group; 0%, 1.2%, 1.5% and 2.1% in the PAGE-B moderate-risk group; and 5%, 11.7%, 12.5%, and 15% in the PAGE-B high-risk group, respectively (log-rank P < 0.001). The AUROCs of the PAGE-B score in the prediction of HCC development at 1, 3, 5 and 10 years were 0.977, 0.903, 0.903 and 0.865, respectively. In the multivariable analysis, older age, male sex, lower platelet levels, presence of cirrhosis, and absence of alanine aminotransferase normalization at month 6 were associated with HCC development (all P < 0.05). CONCLUSION: The PAGE-B score is a practical tool to predict HCC risk in Turkish patients with CHB and may be helpful to improve surveillance strategies.

Comparison of the Efficacy of Entecavir and Tenofovir in Reducing Hepatocellular Carcinoma Risk in Chronic Hepatitis B Patients: A Real-Life Study in Turkey.

BACKGROUND: It is controversial whether entecavir or tenofovir differs in reducing hepatocellular carcinoma (HCC) risk. We aimed to compare the efficacy of entecavir and tenofovir in reducing HCC risk in chronic hepatitis B (CHB) patients. METHODS: This retrospective study included 607 nucleos(t)ide naive CHB patients who had received entecavir or tenofovir. Patients who developed HCC during the first 12 months of therapy were excluded. Cumulative HCC incidences at years 2, 3, 4, 5 and 10 were compared between entecavir and tenofovir groups. Factors associated with HCC were determined by univariate and multivariate analyses. RESULTS: Nineteen (3.1%) patients developed HCC, 12 (4.8%) in entecavir group and 7 (1.9%) in tenofovir group (P = .045). In the entire cohort, cumulative HCC incidences at years 2, 3, 4, 5 and 10 were 1.8%, 2.9%, 4.4%, 5.2% and 9.9% in entecavir group, and 0.6%, 2.4%, 2.4%, 2.4% and 3.7% in tenofovir group, respectively (log-rank P = .130). In multivariate analysis, age ≥50 years, cirrhosis, decompensated cirrhosis, high GGT and low platelet levels were associated with HCC in the entire cohort. In advanced fibrosis/cirrhosis cohort, cumulative HCC incidences at years 2, 3, 4, 5 and 10 were 4.6%, 7.1%, 8.6%, 12.1% and 15.5% in entecavir group, and 1.8%, 5.6%, 5.6%, 5.6% and 8.5% in tenofovir group, respectively (log-rank P = .267). In multivariate analysis, age ≥50 years, decompensated cirrhosis, high GGT and low platelet levels were associated with HCC in the advanced fibrosis/cirrhosis cohort. CONCLUSION: Entecavir and tenofovir are similarly effective in reducing HCC risk in CHB patients.

Validation of the HCC-RESCUE score to predict hepatocellular carcinoma risk in Caucasian chronic hepatitis B patients under entecavir or tenofovir therapy.

The HCC-RESCUE score was developed to predict hepatocellular carcinoma (HCC) risk in Korean chronic hepatitis B (CHB) patients under entecavir therapy. We aimed to validate the HCC-RESCUE score to predict HCC risk in Caucasian CHB patients under entecavir or tenofovir therapy and to compare the predictive performance of the HCC-RESCUE score with those of the CAMD, PAGE-B and modified PAGE-B (mPAGE-B) scores. The study included 647 nucleos(t)ide analogue-naive noncirrhotic and compensated/decompensated cirrhotic patients who had received entecavir or tenofovir for ≥6 months and did not develop HCC during the first 6 months of therapy. Patients with HCC-RESCUE scores ≤64, 65-84 and ≥85 points were classified into low-, intermediate- and high-risk groups, respectively. The AUROCs of the HCC-RESCUE, CAMD, PAGE-B and mPAGE-B scores to predict HCC risk at 5 years were 0.875, 0.870, 0.866 and 0.880, and those at 10 years were 0.862, 0.845, 0.841 and 0.862, respectively (both p > .05). Cumulative HCC incidences at 5 years were 0.0%, 10.5% and 15.8%, and those at 10 years were 1.4%, 15.5% and 24.9%, respectively, in the low-, intermediate- and high-risk groups based on the HCC-RESCUE score (both log rank p < .001). In the entecavir versus tenofovir cohorts, the AUROCs of the HCC-RESCUE score to predict HCC risk at 5 and 10 years were 0.831 versus 0.898 and 0.803 versus 0.910, respectively (both p > .05). The HCC-RESCUE score accurately predicted HCC risk in Caucasian CHB patients under entecavir or tenofovir therapy. A substantial proportion of patients can be dropped from HCC surveillance by using the HCC-RESCUE score.

Real-world efficacy and safety of Ledipasvir + Sofosbuvir and Ombitasvir/Paritaprevir/Ritonavir ± Dasabuvir combination therapies for chronic hepatitis C: A Turkish experience.

BACKGROUND/AIMS: This study aimed to evaluate the real-life efficacy and tolerability of direct-acting antiviral treatments for patients with chronic hepatitis C (CHC) with/without cirrhosis in the Turkish population. MATERIAL AND METHODS: A total of 4,352 patients with CHC from 36 different institutions in Turkey were enrolled. They received ledipasvir (LDV) and sofosbuvir (SOF)±ribavirin (RBV) orombitasvir/paritaprevir/ritonavir±dasabuvir (PrOD)±RBV for 12 or 24 weeks. Sustained virologic response (SVR) rates, factors affecting SVR, safety profile, and hepatocellular cancer (HCC) occurrence were analyzed. RESULTS: SVR12 was achieved in 92.8% of the patients (4,040/4,352) according to intention-to-treat and in 98.3% of the patients (4,040/4,108) according to per-protocol analysis. The SVR12 rates were similar between the treatment regimens (97.2%-100%) and genotypes (95.6%-100%). Patients achieving SVR showed a significant decrease in the mean serum alanine transaminase (ALT) levels (50.90±54.60 U/L to 17.00±14.50 U/L) and model for end-stage liver disease (MELD) scores (7.51±4.54 to 7.32±3.40) (p<0.05). Of the patients, 2 were diagnosed with HCC during the treatment and 14 were diagnosed with HCC 37.0±16.0 weeks post-treatment. Higher initial MELD score (odds ratio [OR]: 1.92, 95% confidence interval [CI]: 1.22-2.38; p=0.023]), higher hepatitis C virus (HCV) RNA levels (OR: 1.44, 95% CI: 1.31-2.28; p=0.038), and higher serum ALT levels (OR: 1.38, 95% CI: 1.21-1.83; p=0.042) were associated with poor SVR12. The most common adverse events were fatigue (12.6%), pruritis (7.3%), increased serum ALT (4.7%) and bilirubin (3.8%) levels, and anemia (3.1%). CONCLUSION: LDV/SOF or PrOD±RBV were effective and tolerable treatments for patients with CHC and with or without advanced liver disease before and after liver transplantation. Although HCV eradication improves the liver function, there is a risk of developing HCC.

AST-platelet ratio index in the prediction of significant fibrosis and cirrhosis in patients with chronic hepatitis B.

BACKGROUND/AIMS: We aimed to evaluate the diagnostic accuracy of AST-platelet ratio index in the prediction of significant fibrosis and cirrhosis in chronic hepatitis B patients by comparison with liver biopsy. MATERIALS AND METHODS: We retrospectively reviewed our computerized data of chronic hepatitis B patients who attended the Gastroenterology Clinic from 2004-2009. Treatment-naive chronic hepatitis B patients who had undergone liver biopsy were included in this study. The degree of fibrosis was scored according to the Ishak staging system. Significant fibrosis was defined as F3-6 and cirrhosis as F5-6. AST-platelet ratio index was calculated based on the original studies. Tests results were compared between the groups F0-2 versus F3-6 and F0-4 versus F5-6. RESULTS: Two hundred and fifty consecutive patients with chronic hepatitis B were included in this study. The area under the ROC curves of AST-platelet ratio index to predict significant fibrosis and cirrhosis were 0.779 and 0.781, respectively. Using cut-off values ≤0.5 and >1.5, significant fibrosis was excluded with a negative predictive value of 91.30% and sensitivity of 87.69% and predicted with a positive predictive value of 59.52% and specificity of 90.81% in 53.60% of patients. Using cut-off values ≤1 and >2, cirrhosis was excluded with a negative predictive value of 92.09% and sensitivity of 64.10% and predicted with a positive predictive value of 33.33% and specificity of 91.47% in 81.60% of patients. CONCLUSIONS: AST-platelet ratio index may be a useful noninvasive marker in the exclusion of both significant fibrosis and cirrhosis in patients with chronic hepatitis B. However, it is not accurate in the prediction of either significant fibrosis or cirrhosis.

Comparison of the efficacy of entecavir and tenofovir in chronic hepatitis B.

BACKGROUND/AIMS: This study aimed to compare the efficacy of entecavir and tenofovir in nucleos(t)ide-naive chronic hepatitis B patients after 48 weeks of therapy. METHODOLOGY: We retrospectively reviewed our data of chronic hepatitis B patients. Nucleos(t)ide-naive patients who had received entecavir or tenofovir for at least 48 weeks were included. We compared entecavir and tenofovir after 48 weeks of therapy with respect to virological, biochemical, serological and histological responses. RESULTS: Of the 44 patients, 24 received entecavir and 20 received tenofovir. Pretreatment characteristics of the patients were similar. After 48 weeks, serum HBV DNA levels decreased by 6.93±1.54log copy/ mL in the entecavir group and 6.89±1.22log copy/mL in the tenofovir group (p=0.65). A similar proportion of patients in entecavir and tenofovir groups achieved undetectable serum HBV DNA (87.5% vs. 95%, p=0.39) and serum ALT normalization (79.2% vs. 85%, p=0.62). The mean histological activity index score improved by 3.83±3.51 points in the entecavir group and 2.20±1.91 points in the tenofovir group (p=0.07), and the mean fibrosis scores improved by 0.38±1.61 points in the entecavir group and 0.70±1.17 points in the tenofovir group after 48 weeks (p=0.44). CONCLUSIONS: Entecavir and tenofovir are similarly effective in nucleos(t)ide-naive chronic hepatitis B patients with high viral load and/or high fibrosis scores after 48 weeks of therapy.

Efficacy of vitamins supplementation to therapy on Helicobacter pylori eradication in patients with low antioxidant capacity.

BACKGROUND AND OBJECTIVES: It was shown that supplementation of vitamins C and E to therapy increased Helicobacter pylori eradication rate. In the present study, we aimed to evaluate whether supplementation of antioxidant vitamins to therapy increases H. pylori eradication rates in patients with chronic stress and low antioxidant capacity. METHODS: This study included 120 patients who presented to gastroenterology outpatient clinic with H. pylori-positive nonulcer dyspepsia and low total antioxidant capacity. Patients in group A (n=80) were given lansoprazole (30 mg, BID), amoxicillin (1000 mg, BID), and clarithromycin (500 mg, BID) for 14 days, as well as vitamin C (500 mg, BID) and vitamin E (200 IU, BID) for 30 days. Patients in group B (n=40) were given lansoprazole (30 mg, BID), amoxicillin (1000 mg, BID), and clarithromycin (500 mg, BID) for 14 days. RESULTS: Total antioxidant capacity were lower than normal levels in all patients. One hundred and fifteen patients (77 in group A, 38 in group B) were analyzed with per protocol analysis. In group A, H. pylori eradication was achieved in 63.8% of the patients included in the intention to treat analysis and in 66.2% of the patients included in the per protocol analysis. In group B, H. pylori eradication was achieved in 42.5% of the patients included in the intention to treat analysis and in 44.7% of the patients included in the per protocol analysis. Eradication rates were significantly higher in group A than in group B (P<0.005). CONCLUSIONS: Supplementation with vitamins C and E increased H. pylori eradication rate of standard triple therapy.

AST-platelet ratio index, Forns index and FIB-4 in the prediction of significant fibrosis and cirrhosis in patients with chronic hepatitis C.

BACKGROUND/AIMS: The aim of this study was to evaluate the diagnostic accuracy of aspartate aminotransferase-platelet ratio index, the Forns index and FIB-4 for the assessment of hepatic fibrosis in chronic hepatitis C patients by comparison with liver biopsy. METHODS: We retrospectively reviewed our computerized data of chronic hepatitis C patients who admitted to the Gastroenterology Clinic between 2004 and 2008. Treatment-naive chronic hepatitis C patients who had undergone liver biopsy and had laboratory test results allowing the calculation of aspartate aminotransferase-platelet ratio index, the Forns index and FIB-4 were included in this study. The degree of fibrosis was scored according to the METAVIR staging system. Significant fibrosis was defined as F2-4 and cirrhosis as F4. Aspartate aminotransferase-platelet ratio index, the Forns index and FIB-4 were calculated based on the original studies. Tests results were compared between groups F0-1 (no or mild fibrosis) versus F2-4 (significant fibrosis) and F03 (no cirrhosis) versus F4 (cirrhosis). RESULTS: One hundred and fifty patients with chronic hepatitis C were included in this study. The areas under the ROC curves of the Forns index, aspartate aminotransferase-platelet ratio index and FIB-4 to predict significant fibrosis were 0.795, 0.774 and 0.764, respectively. The area under the ROC curves of the Forns index, aspartate aminotransferase-platelet ratio index and FIB-4 to predict cirrhosis were 0.879, 0.839 and 0.874, respectively. CONCLUSIONS: The Forns index, aspartate aminotransferase-platelet ratio index and FIB-4 were accurate noninvasive blood tests to predict the presence or absence of significant fibrosis and cirrhosis in half of the chronic hepatitis C patients. The Forns index was slightly better than the aspartate aminotransferase-platelet ratio index and FIB-4 in the prediction of significant fibrosis and cirrhosis.

Serum levels of prostate specific antigen are elevated after colonoscopy.

The risks of prostate cancer and colorectal carcinoma increase with age. So, colonoscopy and measurement of serum prostate specific antigen (PSA) may be performed during a short term in a given patient. We aimed to evaluate whether colonoscopy affects serum PSA levels and to evaluate the relationship between prostate volume and elevation in serum PSA levels after colonoscopy. This study included 44 consecutive male patients, who underwent colonoscopy. The mean age of the patients was 56.05±9.27 years. The mean time required for colonoscopy was 30 min. Serum PSA levels were measured 48-72 hours before colonoscopy, immediately after performing laxative enema, and at 24-48th hour, the 7th day, and the 14th day after colonoscopy in each patient. The serum PSA level was elevated after enema and at 24-48th hour and 7th day after colonoscopy from the baseline (p<0.05), and declined to the baseline by 14th day. When the cut off value of 20 cm3 for normal prostate volume was taken into account, the serum PSA levels were significantly higher at the 24-48th hour and the 7th day in patients with larger prostate volume (>20 cm3) than those with normal prostate volume (p=0.013 and p=0.009). These results suggest that PSA is easily released by manipulations from the larger prostate. In conclusion, serum PSA levels were elevated during 7 days after colonoscopy. Before performing invasive procedures, patients with high serum PSA levels should be asked whether colonoscopy was performed prior to the measurement.

Liver biopsy: ultrasonography guidance is not superior to the blind method.

AIM: The aim of this study was to evaluate our experience with adequate liver biopsy samples and compare the complication rates of blind and US-guided biopsies, as well as to compare the histological yield of blind and US-guided biopsy specimens. METHODS: We retrospectively analyzed 205 consecutive patients that underwent liver biopsies during a 12-month period. Liver biopsy was performed via the blind method in 152 patients, and via US-guidance in 53 patients. Biopsy specimens were evaluated according to length, presence of fragmentation, crush artifacts, adequacy for diagnosis, and the number of portal tracts and central veins. We also evaluated the rates of mortality and major life-threatening complications. RESULTS: All the biopsy specimens were adequate for histological evaluation, except in 8 cases, of which 4 were in the blind biopsy group (2.63%) and 4 were in the US-guided biopsy group (7.54%) (P>0.05). There were no statistically significant differences between the two groups in terms of the specimen fragmentation, or number of portal tracts and central veins in each specimen. Mean specimen length in the US-guided liver biopsy group was 12.58+/-5.59 mm, and in the blind biopsy group 16.22+/-9.91 mm (P<0.005) . There was no mortality or major complications in either of the two study groups. CONCLUSION: US-guided biopsy was not superior to blind biopsy, an unexpected result. Gastroenterologists/hepatologists should be encouraged to perform liver biopsies via the blind method.

Glutaraldehyde-induced colitis: three case reports.

Glutaraldehyde (2% solution) is an effective and widely used disinfecting solution for cold sterilization of endoscopic instruments. Direct contact of glutaraldehyde solution with colonic mucosa can cause self-limited colitis. As it rarely occurs as a complication of colonoscopy, glutaraldehyde-induced colitis is generally reported only as case reports in the literature. We report three cases of glutaraldehyde-induced colitis after colonoscopy. All lesions resolved with supportive treatment. We stress the need for thorough rinsing of the surface and channels of the endoscope with water to avoid the occurrence of this complication.