Cytokeratin-18 Fragments Research Articles

Hepatocellular damage and inflammation in various forms of alcoholic liver disease

The aim of the study was to evaluate hepatocellular damage and immune inflammation in various forms of alcoholic liver disease (ALD). 104 patients with ALD were examined: 15 (14.4%) with liver steatosis (LS), 19 (18.3%) with steatohepatitis and 70 (67.3%) with liver cirrhosis (LC); men 50 (48.1%), women 54 (51.9%); age 45.78.4 years. Traditional clinical, laboratory, instrumental studies were performed, the levels of fragments of cytokeratin-18 (FCK-18), cytokines IL-1, TNF-, IL-4, IL-6, IL-8 were determined by ELISA. The control group consisted of 39 healthy individuals: men 20 (51.2%), women 19 (48.7%), age 48.58.3 years. In LS, an increase in the level of FCK-18 was noted with normal aminotransferase activity, the content of TNF-, IL-6, IL-1, IL-8 increased and the level of IL-4 decreased compared to those in healthy individuals. In steatohepatitis, a triple increase in aminotransferases and FCK-18 was observed compared with LS, as well as an increase in the level of inflammatory mediators, to a greater extent IL-6, to a lesser extent IL-8, TNF-, a decrease in IL-4, IL-1 remained at the same level. In LC, there was a further increase in FCK-18, significantly more pronounced than an increase in AST, and the increase in cytokines continued to the same extent, the levels of IL-6 and IL-8, to a lesser extent IL-1 and TNF-, and the level of IL-4. With the progression of ALD from LS to steatohepatitis, hepatic cell damage was carried out by equally pronounced processes of hepatocyte necrosis and apoptosis, with the development of cirrhosis of the liver, parenchyma damage occurred mainly due to hepatocyte apoptosis. The immuno-inflammatory process progressively increased from the stage of LS to LC with IL-6 and IL-8 undergoing the greatest dynamics. FCK-18 can serve as a non-invasive marker of hepatic cell damage, and IL-6 and IL-8 markers of immune inflammation in ALD.

Pleiotropic effects of ursodeoxycholic acid in patients with non-alcoholic steatohepatitis with impaired glycemic control

The goal was to determine the effect of ursodeoxycholic acid (UDCA) in non-alcoholic steatohepatitis (NASH) with impaired glycemic control.Materials and methods. 67 patients NASH were examined: prediabetes — 38 (56.7%), type 2 diabetes mellitus 29 (43.3%), men — 39 (58.2%), women — 28 (41.8%), age — 45.1 ± 10.2 years. The UDCA dose was 9.4 ± 2.0 mg / kg / day during 59.7 ± 77.6 weeks. Fragments of cytokeratin-18 (FCK-18) (TPS ELISA, Biotech, Sweden), TNF-α (Human TNFα Platinum ELISA, eBioscience, Austria), IL-6 (“Interleukin-6-IFA-Best”, Vector-Best, Russia), insulin (“Insulin TEST System”, Monobind Inc., USA), HOMA-IR were determined.Results. There was a decrease in the levels of FCK-18 — from 238.1 ± 93.7 to 170.7 ± 79.2 U / l (p <0.05), ALT — 61.3 ± 19.0 to 38.9 ± 19.1 U / l (p <0.05), glucose 5.9 ± 1.3 to 5.5 ± 0.7 mmol / l (p <0.05), insulin 21.9 ± 18.2 to 13.7 ± 9.7 MkU / l, HOMA-IR — 5.8 ± 2.2 to 3.1 ± 0.8 (p <0.05), cholesterol — 6.2 ± 0.9 to 5.3 ± 0.3 mmol / l, LDL — 3.9 ± 0.9 to 3.2 ± 0.6 mmol / l (p <0.05), TNF-α 6.3 ± 1.5 to 5.4 ± 2.1 pg / ml (p <0.05), IL-6–7.1 ± 3.4 to 4, 1 ± 3.2 pg / ml (p <0.05).Conclusion. UDCA had pleiotropic effects in NASH with impaired glycemic control, reducing cellular apoptosis, necrosis, inflammation, improving insulin sensitivity and lipid homeostasis.

CYTOKERATIN-18 AS A MARKER OF NON-ALCOHOLIC FATTY LIVER DISEASE IN OBESE ADOLESCENTS

In parallel with the obesity epidemic in the world, the prevalence of non-alcoholic fatty liver disease among children and adolescents is growing. Current data suggest that insulin resistance is one of the main factors in the pathogenesis of non-alcoholic fatty liver disease, and the content of fragments of caspase-cleaved cytokeratin-18 in the blood serum may be one of the informative indicators of non-alcoholic fatty liver disease progression. The aim. To determine mechanisms of formation and progression of non-alcoholic fatty liver disease in obese children and adolescents by evaluating the level of cytokeratin-18. Materials and methods. The study involved 46 adolescents with obesity and non-alcoholic fatty liver disease aged 12–17 years: 19 boys (41.3 %) and 27 girls (58.7 %). Clinical (weight, height, waist and hip circumference), laboratory (glucose, immunoreactive insulin, lipid metabolism, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transpeptidase, cytokeratin-18) parameters were studied and instrumental examination (abdominal ultrasound) was performed. To assess insulin resistance the triglyceride-glucose index was calculated. Results. Depending on the presence of insulin resistance patients were divided into two groups: 21 (45.7 %) of adolescents with insulin resistance and 25 (54.3 %) of adolescents without insulin resistance. Blood tests in patients with insulin resistance revealed significantly higher levels of total cholesterol, triglycerides, very low-density lipoprotein cholesterol, fasting immunoreactive insulin, cytokeratin-18 and gamma-glutamyl transpeptidase. All adolescents were divided into 2 groups depending on the level of cytokeratin-18: patients with cytokeratin-18 >233 mIU/ml and <233 mIU/ml (15 (32.6 %) and 31 (67.4 %) respectively). It was found that there were significantly more patients with insulin resistance in the group with the level of cytokeratin-18 >233 mIU/ml. Conclusion. In obese adolescents with non-alcoholic fatty liver disease insulin resistance is associated with more pronounced disorders of lipid and carbohydrate metabolism and higher levels of markers that characterize the state of the liver such as cytokeratin-18 and gamma-glutamyl transpeptidase. Adolescents with obesity and non-alcoholic fatty liver disease with a threshold level of cytokeratin-18, which indicates the transformation of steatosis into steatohepatitis, two times more often have present insulin resistance.

THU108 - Plasma protein glycomics combined with circulating fragments of cytokeratin-18 are reliable biomarkers to diagnose alcoholic hepatitis

THU108 - Plasma protein glycomics combined with circulating fragments of cytokeratin-18 are reliable biomarkers to diagnose alcoholic hepatitis

Diagnostic methods and pharmacological management of non-alcoholic fatty liver disease – There is still room for improvement

Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic steatosis in the absence of excessive alcohol consumption and it may progress to non-alcoholic steatohepatitis, cirrhosis, liver failure, and cancer. Despite the alarming rate of the disease observed in the last decades due to the increase in the prevalence of the metabolic syndrome,there are unmet needs in the diagnosis and the management of the disease. The gold standard for the diagnosis of the disease remains the liver biopsy. The ultrasound, the laboratory tests (serum aminotransferases, gamma-glutamyl transpeptidase, and ferritin levels, cytokeratin-18 fragment and fibrosis growth factor 21), the clinical (Fatty Liver Index [FLI], NAFLD fibrosis score [NFS] and Fibrosis 4 calculator [FIB-4]) and the non-invasive scores (NIS-4, FS3) are being developed and evaluated and they are very promising for both detecting the presence and assessing the severity of the disease. For the management of NASH, no pharmacological treatment has been approved. Vitamin E, thiazolidinediones, sodium-glucose transport protein-2 (SGLT-2) inhibitors, inhibitors of dipeptidyl peptidase 4 (DPP-4), glucagon-like peptide-1 receptor agonists (GLP-1 RAs), statins, and many other agents under investigation may have a role in the management of NAFLD alongside their primary indications.

FICK-3 Score Combining Fibrosis-4, Insulin Resistance and Cytokeratin-18 in Predicting Non-alcoholic Steatohepatitis in NAFLD Egyptian Patients.

As liver biopsy had multiple procedure-related complications, the introduction of reliable noninvasive tests for accurate discrimination of NASH and liver fibrosis in patients with NAFLD are mandatory. The aim was to elucidate the diagnostic value of fibrosis-4 index (FIB-4), Homeostasis Model Assessment for Insulin Resistance (HOMA-IR) and Cytokeratin-18 fragments (CK18-Fs) in the prediction of NASH and liver fibrosis. One hundred Egyptian patients with NAFLD selected from outpatient's clinics of Mansoura University Hospitals underwent histological examination through liver biopsy after approval and consent. The FIB-4, HOMA-IR, CK18-Fs (measured using a human ELISA Kit) and their combination in FICK-3 score were investigated for predicting NASH and liver fibrosis. Receiver operating characteristic (ROC) curves and multivariate logistic regression were analyzed. In patients with NAFLD, the areas under the ROC were significantly high (AUC: 0.765, 0.700, 0.803, 0.835 for FIB-4, HOMA-IR, CK18-Fs, FICK-3 score, respectively, p = 0.05) displaying a highly statistically significant predictive ability for NASH. Significantly higher AUCs for these parameters were demonstrated predicting early-or advanced-stage liver fibrosis (AUC >0.7, p<0.01). Also, the combined FICK-3 score (the sum of FIB-4 >1.46, HOMA-IR >2.11 and CK18-Fs >307U L-1) had highly significant predictive values for NASH and liver fibrosis and had the best diagnostic accuracy at a cutoff value of 1(AUC >0.8, p<0.001). Contrasted with other diagnostic scores, FICK-3 had the best diagnostic accuracy for detection of fibrotic NASH (AUC = 0.954, p<0.001) and positively correlated with the histological features of NAFLD. The new combination FICK-3 score was a reliable and significant predictor for NASH and liver fibrosis in NAFLD Egyptian patients.

The effect of total anthocyanin-base standardized (Cornus mas L.) fruit extract on liver function, tumor necrosis factor \u03b1, malondealdehyde, and adiponectin in patients with non-alcoholic fatty liver: a study protocol for a double-blind randomized clinical trial

BackgroundNonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide. Evidence showed that anthocyanins might have effects on NAFLD. Protective effects of Cornelian cherry (Cornus mas L.) extract, as an anthocyanins-rich source, on liver were reported in animal studies. However, very few clinical trials were conducted in this regard. Thus, the aim of this research will be to evaluate the effect of supplementation with total anthocyanin-base standardized cornelian cherry fruit extract on liver function (Serum levels of Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), cytokeratin-18 fragment M30 (CK-18 M30), as well as steatosis and fibrosis of liver), tumor necrosis factor α (TNF-α), malondealdehyde (MDA), and adiponectin in patients with NAFLD.MethodsIn a double-blind randomized clinical trial, 80 NAFLD patients will be studied. The patients will be randomly assigned into two groups. The intervention group will receive the cornelian cherry extract, containing 320 mg.d− 1 anthocyanins, per day for 12 weeks. The control group will also take the placebo daily for 12 weeks. Liver function (Serum levels of AST, ALT and CK-18 M30; steatosis and fibrosis of liver), serum levels of TNF-α, MDA, and adiponectin will be measured at the baseline and the end of trial for both groups and their results will be compared.DiscussionConsidering evidences about the useful impacts of anthocyanins on NAFLD, the effects of supplementation with cornelian cherry extract will be investigated on the important variables related to NAFLD.Trial registrationIranian Registry of Clinical Trials (IRCT20180419039359N1).

Chemerin, retinol binding protein-4, cytokeratin-18 and transgelin-2 presence in sera of patients with non-alcoholic liver fatty disease

Background. Chemerin and retinol binding protein-4 (RBP-4) are adipokines which may play a role in the progression of NAFLD. It has been also suggested that cytokeratin-18 (CK-18) could be a marker of hepatocyte caspase-directed death while transgelin-2 production could reflect stage of liver fibrosis. The aim of this study was to evaluate the level of the above adipokines in sera of patients with NAFLD and determine the relation between the level of transgelin-2 and fibrosis-4 index (FIB-4).Material and methods. Ninety-five subjects included initially to the study were divided into four groups: (I) prediabetics, obese with NAFLD and metabolic syndrome (Ms), (II) lean with NAFLD and without MS, (III) obese without NAFLD and MS, and (IV) healthy individuals. We determined the levels of chemerin, RBP-4, transgelin-2 and CK-18 fragments in sera of patients with NAFLD. Moreover, we examined if the levels of CK-18 fragments and transgelin-2 correlates with FIB4 value. Results. Chemerin and RBP-4 were highly expressed in sera of all NAFLD, especially in obese individuals. Chemerin level was also linked to MS. High level of serum CK-18 fragments and transgelin-2 did not correlate with obesity and MS, but seemed to correlate with progression of NAFLD to liver fibrosis.Conclusions. In conclusion, the production of the two adipokines, chemerin and RBP-4, is strongly associated with obesity in patients with NAFLD. Serum concentrations of CK-18 fragments and transgelin-2 correlate with the severity of NAFLD, but not with obesity.

Epithelial cell death markers in bronchoalveolar lavage correlate with chronic lung allograft dysfunction subtypes and survival in lung transplant recipients-a single-center retrospective cohort study.

Chronic lung allograft dysfunction (CLAD) remains the leading cause of late death after lung transplantation. Epithelial injury is thought to be a key event in the pathogenesis of CLAD. M30 and M65 are fragments of cytokeratin-18 released specifically during epithelial cell apoptosis and total cell death, respectively. We investigated whether M30 and M65 levels in bronchoalveolar lavage (BAL) correlate with CLAD subtypes: restrictive allograft syndrome (RAS) versus bronchiolitis obliterans syndrome (BOS). BALs were obtained from 26 patients with established CLAD (10 RAS, 16 BOS) and 19 long-term CLAD-free controls. Samples with concurrent infection or acute rejection were excluded. Protein levels were measured by ELISA. Variables were compared using Kruskal-Wallis, Mann-Whitney U test and Chi-squared tests. Association of M30 and M65 levels with post-CLAD survival was assessed using a Cox PH models. M65 levels were significantly higher in RAS compared to BOS and long-term CLAD-free controls and correlated with worse post-CLAD survival. Lung epithelial cell death is enhanced in patients with RAS. Detection of BAL M65 may be used to differentiate CLAD subtypes and as a prognostic marker in patients with established CLAD. Understanding the role of epithelial cell death in CLAD pathogenesis may help identify new therapeutic targets to improve outcome.

Treatment of non-alcoholic steatohepatitis patients with vitamin D: a double-blinded, randomized, placebo-controlled pilot study

Background: Non-alcoholic steatohepatitis (NASH) is defined by liver inflammation and consecutive fibrotic damage caused by a deposition of fat in the liver. No licensed medical treatments exist and lifestyle modification is difficult to incorporate into everyday life. We investigated the efficacy and safety of a 48-week treatment with vitamin D3 in NASH patients.Methods: Histologically determined NASH patients with elevated alanine aminotransferase (ALT) and decreased 25-OH vitamin D level at baseline received vitamin D3 or placebo orally over a 48-week period. The primary endpoint of this study was the change in ALT from baseline to the end-of-treatment. Steatohepatitis was categorized according to the Steatosis, Activity and Fibrosis Score and disease activity was assessed using the NAFLD activity score.Results: Serum 25-OH vitamin D levels significantly increased only in the vitamin D3 group over the 48-week treatment phase indicating compliance. In contrast to placebo, patients in the vitamin D group had markedly decreased ALT levels after the end-of-treatment phase. A significant decrease during treatment with vitamin D was also observed for cytokeratin-18 fragments compared with placebo. The study was not powered to detect changes in histological score, hence only descriptive results for histopathological characteristics are available.Conclusions: Treatment with 2100 IE vitamin D q.d. over 48 weeks was well tolerated and led to a significant improvement of serum ALT levels in patients with hypovitaminosis D and histology-proven NASH as the primary endpoint together with a trend toward reduction of hepatic steatosis, which was not significant due to a small number of available biopsy specimens.

The clinical significance of insulin resistance in non-diabetic patients with early forms of non-alcoholic fatty liver disease.

To assess the presence of insulin resistance (IR) in non-diabetic patients with early forms of non-alcoholic fatty liver disease (NAFLD) - liver steatosis (LS) and steatohepatitis (SH) of mild activity and the influence of IR on the clinical course of these diseases. 134 patients with NAFLD were examined: 54 with LS and 80 with SH. The control group consisted of 37 healthy donors. Anthropometric parameters (body mass index (BMI), waist circumference (WC)), clinical and biochemical blood indices, including the blood level of cytokeratin-18 fragments (CK-18), TNF-α and IL-6 cytokines, insulin were evaluated. The HOMA index and the fibrosis index (NAFLD FS) were calculated. Patients were divided into groups: I - with the absence of IR (HOMA-index &lt;2.7), II - with the presence of IR (HOMA-index&gt; 2.7). Indicators of hepatic injury, inflammation, cholestasis, fibrosis and atherogenic dyslipidemia are higher in patients with LS of group II (with IR) than in group I patients (without IR). BMI, WC, γ-glutamil transpeptidase, CK-18 and fibrosis index are significantly higher in group II patients with SH compared with group I, there is no significant difference in the level of cytolysis, inflammation and dyslipidemia indices. A high incidence of IR in non-diabetic patients with LS (37.0%) and SH (55.0%) was found and the effect of IR on the clinical course of these diseases was revealed. Insulin resistance in non-diabetic patients with NAFLD was detected in SH (55.0%) with higher frequency than in LS (37.0%). In LS, IR is associated with impaired hepatic cell damage, intrahepatic cholestasis, atherogenic dyslipidemia and fibrosis. In SH, IR is combined with reliable growth in indicators of hepatocyte apoptosis, cytokine proinflammatory status and fibrosis. IR determines the progressing course of NAFLD, promoting the transformation of steatosis into steatohepatitis and steatohepatitis into fibrosis and liver cirrhosis.

Changes in serum levels of cytokeratin-18 fragments in patients with chronic hepatitis C under antiviral therapy

Background and Aims: Cytokeratin-18 is the known substrate for caspases, which are encountered during hepatic and pancreatic acinar apoptosis. Studies performed in recent years have indicated that the cleavage level of serum cytokeratin-18 (M30 antigen) is correlated with hepatic fibrosis and disease severity in both chronic hepatitis C and non-alcoholic steatohepatitis. It was shown that antiviral therapy in chronic viral hepatitis C patients significantly reduced hepatocellular apoptosis and cytokeratin-18 is accepted as a reliable marker of hepatocyte apoptosis. Our aim was to determine the correlation between the cytokeratin-18 level and treatment response in patients with chronic viral hepatitis C. Materials and Methods: Sixty patients with chronic viral hepatitis C were included in the study. A 48-week course of peginterferon- ribavirin therapy was given to appropriate patients. Hepatitis C virus RNA was measured at 0, 12, and 24 weeks at the end of therapy and 72 weeks. In addition, cytokeratin-18 levels were measured at 0, 24, and 72 weeks. Results: The mean age of 60 patients was 52±10.9 years. While 31 (51.6%) of patients were in the sustained viral response group, 29 (8.4%) of patients were in the non-sustained viral response group. It was determined that while the cytokeratin-18 level at week 0 in the sustained viral response group was 243±21, the cytokeratin-18 level at week 24 was 115±12 U/L and the difference between the level of cytokeratin-18 at weeks 0 and 24 were 127±209 U/L (p: .014). While the cytokeratin-18 level at week 0 in the non- sustained viral response group was 270±14; at week 24, the cytokeratin-18 level was 136±19 U/L and the difference between cytokeratin-18 levels at weeks 0 and 24 was 136±156 U/L (p &amp;gt; .5). At week 72, the cytokeratin-18 level in the sustained viral response group was 109±38 and the difference between weeks 0 and 72 was 134±215 (p &amp;lt; .002). Conclusion: In chronic viral hepatitis C patients, there was a correlation between sustained viral response and cytokeratin-18, which is a marker of apoptosis. During treatment, it was found that there was a relationship between sustained viral response and the decrease in cytokeratin-18 levels. Thisfinding indicates that cytokeratin-18 level monitoring may be used as a predictive marker of sustained viral response.

Intrahepatic cholestasis in nonalcoholic fatty liver disease.

To determine the frequency of intrahepatic cholestasis and its impact on the clinical features of the different forms of non-alcoholic fatty liver disease (NAFLD). The study involved 163 patients with NAFLD: 92 (56.4%) with hepatic steatosis (HS), 56 (34.4%) with steatohep- atitis (SH) and 15 (9.2%) with liver cirrhosis (LC). Diagnosis is based on clinical, laboratory, ultrasound and histological data. Insulin, tu- mor necrosis factor α (TNF-α), fragments of cytokeratin-18 (FCK-18) were determined by ELISA. The index of insulin resistance (HOMA- IR) was calculated. NAFLD fibrosis score (NAFLD-FS) was determined, taking into account the patient's age, body mass index, presence or absence of carbohydrate metabolism disturbances, levels of ASAT, ALAT, albumin and blood platelets. Cholestatic syndrome was detected in 49 (30.1%) NAFLD patients: in 23 (25%) with HS, in 19 (33.9%) with SH and in 7 (46.7%) with LC. Patients with HS, SH and LC with signs of cholestasis as compared to patients with the same forms of NAFLD without cholestasis had significantly higher levels of the following indicators: aminotransferases, triglycerides, HOMA-IR, TNF-α, FCK-18, NAFLD-FS, - the number of platelets is reduced, indirectly confirming the more rapid development of fibrosis in cholestasis. These findings were consistent with published data on the violation in cholestasis regulatory functions of bile acids, which are ligands of hepatocyte nuclear receptor, re- sponsible for normal homeostasis. In all forms of NAFLD with cholestasis were detected more pronounced liver cell inflammation, hepatocyte necrosis and apoptosis, fibrosis, disturbance of carbohydrate and lipid metabolism, which contributed to a progressive course of NAFLD and confirmed the need for medical correction of cholestasis, starting with the earliest form of NAFLD - hepatosteatosis.

Immunomonitoring of MSC-Treated GvHD Patients Reveals Only Moderate Potential for Response Prediction but Indicates Treatment Safety

Mesenchymal stromal cells (MSCs) are used as salvage therapy to treat steroid-refractory acute graft-versus-host disease (aGvHD). We studied the immunological response to MSC treatment in 16 aGvHD patients by assessing lymphocyte profiles and three proposed aGvHD serum markers during the MSC treatment. Surprisingly, there were no obvious differences in the lymphocyte profiles between the responders and non-responders. The numbers of T, B, and NK cells were below the normal reference interval in all patients. CD4+ T helper (Th) cell levels remained particularly low throughout the follow-up period. The relative proportion of Th1 cells decreased, while regulatory T cells remained unaltered, and only very few Th2 and Th17 cells could be detected. Serum concentrations of regenerating islet-derived protein 3-alpha, cytokeratin-18 fragments (CK18F), and elafin were significantly elevated in patient samples compared with healthy controls, but only CK18F showed any potential in the prediction of patients’ response to MSCs. No obvious markers for MSC therapy response were revealed in this study, but the results suggest that allogeneic MSCs do not provoke overt T cell-mediated immune responses at least in immunosuppressed aGvHD patients. The results advocate for the safety of MSC therapy and bring new insights in MSC immunomodulation mechanisms.

Serum levels of a cell death biomarker predict the development of cirrhosis-related conditions in primary biliary cholangitis.

Non-invasive predictors for the development of cirrhosis-related conditions are needed for patients with primary biliary cholangitis (PBC). We investigated the association between cytokeratin-18 fragments (M30 and M65) and liver histology, treatment response and the development of cirrhosis-related conditions in patients with PBC. We retrospectively reviewed the clinical data of 111 individuals with biopsy-proven PBC. Serum M30 and M65 levels were measured using stored sera. M30 were significantly decreased after treatment, but there was no significant change in the M65 levels. M65 was significantly higher in non-responders according to the Paris-I and Paris-II definitions. In the multivariate analysis, high levels of M65 were significantly associated with advanced Scheuer stage (odds ratio 5.86; 95% confidence interval 0.55-22.2; P = 0.009) and with the development of cirrhosis-related conditions (hazard ratio 3.94; 95% confidence interval: 1.06-14.5, P = 0.039). Among PBC patients without cirrhosis, those with high serum M65 levels at baseline were at higher risk of developing cirrhosis-related conditions (log-rank test; P = 0.001). High levels of serum M65 may be a non-invasive and early predictor of the development of cirrhosis-related conditions in PBC patients. Our findings may help initiate therapies earlier for those at risk for cirrhosis.

New scoring system combining the FIB-4 index and cytokeratin-18 fragments for predicting steatohepatitis and liver fibrosis in patients with nonalcoholic fatty liver disease

Purpose: To establish a new scoring system as a noninvasive tool for predicting steatohepatitis and liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD).Methods: A total of 170 patients histologically diagnosed with nonalcoholic steatohepatitis (NASH) (n = 130) or nonalcoholic fatty liver (NAFL) (n = 40) were enrolled. We analyzed receiver operating characteristic (ROC) curves and performed multivariate analysis to predict steatohepatitis and liver fibrosis.Results: Multivariate analysis showed that cytokeratin-18 fragment (CK18-F) levels (≥278 U/L) (odds ratio [OR], 4.46; 95% confidence interval [CI], 1.42–14.00; p = 0.010) and the FIB-4 index (≥1.46) (OR, 4.54; 95% CI, 1.93–29.50; p = 0.004) were independently associated with prediction of NASH. We then established a new scoring system (named the FIC-22 score) for predicting NASH using CK18-F levels and FIB-4 index. The areas under the ROC curve (AUROCs) of the FIC-22 score and NAFIC score were 0.82 (95% CI, 0.75–0.89) and 0.71 (95% CI, 0.62–0.78) (p = 0.044). Additionally, the AUROC of the FIC-22 score for predicting the presence of fibrosis (F ≥ 1) was 0.78 (95% CI, 0.70–0.85).Conclusions: In patients with NAFLD, the FIC-22 score had high predictive accuracy not only for steatohepatitis but also for the presence of liver fibrosis.

Serum cytokeratin-18 is a non-invasive biomarker for evaluating disease severity in patients with liver cirrhosis and hepatocellular carcinoma

Background: To investigate the dynamic changes in cytokeratin-18 (CK18) levels in patients with liver cirrhosis and hepatocellular carcinoma (HCC) undergoing surgery, and to evaluate its diagnostic value for HCC and acute liver injury. Methods: Serum levels of total CK18 and CK18 fragments were measured using specific enzyme-linked immunosorbent assays in patients with liver cirrhosis and HCC, and control patients with liver cavernous hemangioma. Clinical parameters were measured 1 day before and 1, 3, and 5 days after surgery. Results: CK18 levels were markedly elevated in patients with liver cirrhosis and HCC compared with control patients with non-cirrhotic liver disease (total CK18: 262.9±130.0 U/L, control 158.7±44.5 U/L, P=0.038; CK18 fragments: 145.1±69.6 U/L, control 81.6±15.2 U/L, P=0.005), but fell to near preoperative levels at postoperative day (POD) 5. CK18 levels paralleled perioperative changes in alanine transaminase (ALT), reaching peak values on POD 1, and then decreasing gradually to near preoperative levels by POD 5. The correlation coefficients for the relationships between serum CK18 released by hepatocyte necrosis and ALT levels were 0.659 in the cancer group and 0.922 in the control group. Serum ALT levels remained above preoperative levels at POD 5. The levels of total CK18 and CK18 fragment released by hepatocyte necrosis were remarkably higher on POD 1 than the levels before operation. Conclusions: Serum levels of total CK18 and CK18 fragments were higher in HCC patients with chronic hepatitis B-related liver cirrhosis compared with levels in patients with liver cavernous hemangioma. Liver cancer may have a greater impact on serum CK18 levels than cirrhosis. Surgical injury can increase serum CK18 levels, and CK18 may be a more sensitive marker of liver injury than ALT.

The use of cytokeratin-18 fragments in diagnostics of non-alcohol fatty liver disease

Aim. To evaluate the value of cytokeratin-18 fragments (CKF-18) for diagnostics of non-alcohol fatty liver disease (NAFLD): liver steatosis (LS) and steatohepatitis (SH). Material and methods. We examined 148 patients with NAFLD (94 men, 49 women) including 28 (19.6%) with LS and 115 (80.4%) with SH (mean age 48.85±10.36 and 48.6±12.0 years respectively). The diagnosis was verified based on clinical, ultrasonic, and liver biopsy (in 17 (12.0%) patients) data. CKF-18 were measured by the immunoenzyme assay (TPS ELISA,Biotech, Sweden) and TNF-alpha with the use of a Human TNFα Platinum ELISA kit (Bioscience, Austria). Control group consisted of 27 healthy donors. Results. CKF-18 level in SG patients (420.5±426.9 U/l) was significantly higher than in LS patients and healthy subjects (126.13±48.0 and 129.27±59.78 U/l respectively). TNF-alpha levels were 6.28±0.5, 5.15±0.6, and 4.81±0.7 pg/ml respectively (p&lt;0,05). Correlation of CKF-18 levels with the following parameters was documented : ALT (r=+0,45, p&lt;0,05); AST (r=+0,43, p&lt;0,05), triglycerides (r=+0,25, p&lt;0,05), HDLP (r=-0,21, p&lt;0,05), histological activity index (r=+0,46, p&lt;0,05). No correlation was found between CKF-18 and TNF-alpha levels or between CKF-18 and severity of fibrosis. Conclusion. CKF-18 levels in SH patients are significantly higher than in LF patients and correlate with histological signs of SH activity. Measurement of TNF-alpha can be used for differential diagnostics of SH and LF. Significant elevation of aminotransferase activities and CKF-18 levels in SH compared with LF and their close correlation confirms the important role of necrosis and hepatocyte apoptosis in pathogenesis of NAFLD.

Ability of Cytokeratin-18 Fragments and FIB-4 Index to Diagnose Overall and Mild Fibrosis Nonalcoholic Steatohepatitis in Japanese Nonalcoholic Fatty Liver Disease Patients

Background: Several laboratory markers used in lieu of liver biopsy are reportedly useful in the diagnosis of nonalcoholic steatohepatitis (NASH). In the present study, we investigated the diagnostic impact of various non-invasive markers for predicting NASH. Methods: A total of 229 nonalcoholic fatty liver disease (NAFLD) patients who underwent liver biopsy were enrolled for the study. The diagnostic ability of various markers to diagnose NASH from NAFLD was investigated. Results: A total of 140 patients were histologically diagnosed with NASH. Of these, 104 had degree 0-2 fibrosis (F0-2), and 36 had degree 3-4 fibrosis (F3-4). Multiple logistic regression analysis identified hyaluronic acid (HA) (OR 1.014; 95% CI 1.002-1.026; p = 0.024), FIB-4 index (OR 2.097; 95% CI 1.177-3.735; p = 0.012), and cytokeratin-18 fragments (CK-18F) (OR 1.002; 95% CI 1.001-1.002; p < 0.001) as factors independently associated with the diagnosis of NASH. The areas under the receiver operating characteristic curves (AUROCs) of HA, FIB-4 index, and CK-18F for the diagnosis of NASH were 0.77, 0.76, and 0.72, respectively. In addition, FIB-4 index (OR 1.907; 95% CI 1.063-3.419; p = 0.03) and CK-18F (OR 1.002; 95% CI 1.001-1.002; p < 0.001) could differentiate between NASH and NAFL, even when NASH patients with advanced fibrosis (F3-4) were excluded. AUROCs of FIB-4 index and CK-18F for the diagnosis of NASH with mild fibrosis (F0-2) from NAFLD were 0.70 and 0.70, respectively. Conclusions: FIB-4 index and CK-18F have good diagnostic abilities not only for NASH overall, but also for NASH with mild fibrosis.

Complementary Role of Fibroblast Growth Factor 21 and Cytokeratin 18 in Monitoring the Different Stages of Nonalcoholic Fatty Liver Disease

Fibroblast growth factor 21 (FGF21) and cytokeratin 18 (CK18) were previously reported to be elevated in nonalcoholic fatty liver disease (NAFLD). We aim to analyze the differential roles of FGF21, cell apoptosis marker CK18 fragment M30 and total cell death marker CK18 M65ED in monitoring the different stages of NAFLD spectrum in a population-based prospective cohort comprising 808 Chinese subjects. Predictive performances for monitoring the different stages of NAFLD were assessed by logistic regression and receiver-operating characteristic (ROC) curves. We found baseline FGF21 but not CK18 level was an independent predictor for the development of simple steatosis. NAFLD patients who had remission during follow-up had significantly lower baseline M30 levels than those who sustained NAFLD (84.74U/L [53.26–135.79] vs. 118.47U/L [87.16–188.89], P = 0.012). M65ED was independently predictive of progressing to suspected non-alcoholic steatohepatitis (NASH) in NAFLD patients. These results suggest that FGF21 can be used for early identification of hepatic steatosis. On the other hand, CK18 including M30 and M65ED, are predictive of the prognosis of NAFLD patients. FGF21 and CK18 might play differential roles and have complementary value in non-invasive identification and monitoring the outcome of NAFLD patients.