Serum M30 Research Articles

Serum Cytokeratin-18 levels as a prognostic biomarker in advanced liver disease: a comprehensive meta-analysis

Cytokeratin-18 (CK-18) is a marker of hepatic cell death. Serum CK-18 could serve as a prognostic marker for patients with advanced liver disease (ALD). This meta-analysis aims to explore the association between total CK-18 (M65) and caspase-cleaved CK-18 (M30) levels with the prognosis of ALD patients. Relevant longitudinal observational studies were identified through comprehensive searches of the Medline, Web of Science, and Embase databases. A random-effects model was utilized to synthesize the findings, accommodating heterogeneity among studies. The analysis included 14 datasets from 11 studies. Elevated serum CK-18 levels at admission were linked to a higher risk of death or liver transplantation during follow-up. This association was consistent for both M65 (risk ratio [RR] 1.99, 95% confidence interval [CI] 1.65 to 2.40, p < 0.001; I2 = 43%) and M30 (RR 1.94, 95% CI 1.57 to 2.40, p < 0.001; I2 = 46%). Subgroup analysis revealed that the relationship between serum M65 levels and adverse outcomes was attenuated in studies using multivariate analysis compared to those using univariate analysis (RR 1.78 vs. 2.80, p for subgroup difference = 0.03). Further subgroup analyses indicated that the prognostic significance of CK-18 for ALD patients was not significantly influenced by study design, methods of determining CK-18 cutoff values, or follow-up durations. Elevated serum CK-18 levels at admission indicate a poor prognosis in patients with ALD. This finding holds for both M65 and M30.

Clinical significance of serum Ck18-M65 and M30 levels in patients with chronic hepatitis B combined with nonalcoholic steatohepatitis and liver fibrosis

In this study, we aimed to explore the clinical significance of serum CK18-M65 and CK18-M30 levels in patients with chronic hepatitis B (CHB) complicated by nonalcoholic steatohepatitis (NASH) and liver fibrosis. The observation and control groups comprised 133 patients with CHB complicated by NASH and 50 healthy patients from our hospital, respectively. Liver function indices, including alanine aminotransferase, glutamic aminotransferase, γ-glutamyltransferase, total bilirubin, total protein, and total cholesterol, were determined using an automatic biochemical analyzer. Hyaluronic acid, type III procollagen, type IV collagen, laminin, and CK18-M65 and M30 levels were detected using ELISA. Serum CK18-M65 and M30 levels in patients with CHB complicated by NASH were positively correlated with the liver fibrosis stage (P &lt; .05). While serum CK18-M65 demonstrated a low diagnostic value for liver fibrosis in the S0-1 stage, it exhibited good diagnostic value for S2-3 stage liver fibrosis. Serum CK18-M30 displayed good diagnostic value for S0-1 and S2-3 hepatic fibrosis, particularly for S2-3 hepatic fibrosis. Elevated serum CK18-M65 and CK18-M30 levels in patients with CHB complicated with NASH suggest their potential utility in evaluating the progression of liver fibrosis in this population. In particular, CK18-M30 exhibits superior diagnostic efficiency.

Can serum M30 levels be utilized as an activation marker in patients with ulcerative colitis?

Ascertainment of disease activation is an important component of therapeutic decisions in ulcerative colitis patients and may present certain clinical challenges. The objective of this study was to determine serum levels of the M30 fragment of cytokeratin 18 and its utility as an activation marker in patients with ulcerative colitis, who are known to have increased apoptosis. A total of 60 ulcerative colitis (30 active and 30 remission) patients aged over 18 years and 29 healthy individuals as controls were included in the study. M30, C-reactive protein, and mean platelet volume were evaluated in all participants and compared between ulcerative colitis patients and controls, as well as between those with active disease or remission. Although ulcerative colitis patients with active disease had higher M30 levels than those in remission, the difference was not statistically significant (p=0.085). The mean M30 levels tended to increase with increasing extent of involvement, although the differences were not significant (p=0.065). The comparison of C-reactive protein and mean platelet volume according to the site of involvement, however, showed significant differences (p=0.02 and 0.004, respectively). M30 did not show significant correlations with C-reactive protein, mean platelet volume, and Mayo Score (p=0.0834, 0.768, and 0.401, respectively). Our results suggest that, in contrast to C-reactive protein and mean platelet volume, M30 levels do not have a significant role as an activation marker in ulcerative colitis patients. Thus, we believe that M30 may not represent an appropriate marker to be used for this purpose.

Serum keratin-18 detects hepatic inflammation and predicts progression in compensated alcohol-associated liver disease.

Alcohol-associated liver fibrosis accumulates over decades, driven by hepatic inflammation and cell death. We investigated the diagnostic accuracy of keratin-18 degradation, measured using serum M30 and M65 levels, and the ActiTest for hepatic inflammatory activity in patients with compensated alcohol-associated liver disease (ALD). Furthermore, we evaluated the prognostic accuracy of markers for liver-related events and all-cause mortality. All findings were compared with routine liver function tests: Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and gamma-glutamyltransferase. Our prospective, biopsy-controlled, single-center study included 265 patients with ongoing or prior excessive alcohol intake, representing the full spectrum of compensated ALD. We defined hepatic inflammatory activity as a combined score of lobular inflammation and ballooning. For severe hepatic inflammatory activity (n=40), we found excellent diagnostic accuracy for M30 (area under the receiver operating characteristics curve [AUROC]=0.90), M65 (AUROC=0.86), and AST (AUROC=0.86). Elevated M30 (M30 > 240 U/L) had the highest positive predictive value (PPV) and specificity, significantly higher than M65, ActiTest and ALT, but not AST (M30: sensitivity=83%, specificity=82%, positive predictive value=45%, negative predictive value=95%). Patients were followed up for 1445 patient-years. All markers, except for ALT, significantly predicted liver-related events and all-cause mortality. After adjusting for advanced fibrosis, drinking behavior and body mass index, M30 and M65 remained significant predictors of liver-related events, whereas M30 and AST were significant predictors of all-cause mortality. Conclusion: M30 and AST accurately detect severe hepatic inflammatory activity in patients with compensated ALD. M30 was the only significant predictor of both liver-related events and all-cause mortality after adjusting for advanced fibrosis, body mass index, and drinking behavior at inclusion.

Clinical usefulness and acceleratory effect of macrophage inhibitory cytokine-1 on biliary tract cancer: an experimental biomarker analysis

BackgroundBiliary tract cancer (BTC) has a poor prognosis; therefore, useful biomarkers and treatments are needed. Serum levels of macrophage inhibitory cytokine-1 (MIC-1), a member of the TGF-β superfamily, are elevated in patients with pancreaticobiliary cancers. However, the effect of MIC-1 on BTC is unknown. Therefore, we investigated the effect of MIC-1 on BTC and assessed whether MIC-1 is a biomarker of or therapeutic target for BTC.MethodsMIC-1 expression in BTC cells was determined by performing histological immunostaining, tissue microarray (TMA), western blotting, and reverse transcription PCR (RT–PCR). Cell culture experiments were performed to investigate the effect of MIC-1 on BTC cell lines (HuCCT-1 and TFK-1). The relationships between serum MIC-1 levels and either the disease state or the serum level of the apoptosis marker M30 were retrospectively verified in 118 patients with pancreaticobiliary disease (individuals with benign disease served as a control group, n = 62; BTC, n = 56). The most efficient diagnostic marker for BTC was also investigated.ResultsMIC-1 expression was confirmed in BTC tissue specimens and was higher in BTC cells than in normal bile duct epithelial cells, as determined using TMA, western blotting and RT–PCR. In cell culture experiments, MIC-1 increased BTC cell proliferation and invasion by preventing apoptosis and inhibited the effect of gemcitabine. In serum analyses, serum MIC-1 levels showed a positive correlation with BTC progression and serum M30 levels. The ability to diagnose BTC at an early stage or at all stages was improved using the combination of MIC-1 and M30. The overall survival was significantly longer in BTC patients with serum MIC-1 < the median than in BTC patients with serum MIC-1 ≥ the median.ConclusionsMIC-1 is a useful diagnostic and prognostic biomarker and might be a potential therapeutic target for BTC.

Non-Invasive Detection of Fibrotic NASH in NAFLD Patients with Low or Intermediate FIB-4.

Background: Non-alcoholic steatohepatitis (NASH) and fibrosis are the main prognostic factors in non-alcoholic fatty liver disease (NAFLD). The FIB-4 score has been suggested as an initial test for the exclusion of progressed fibrosis. However, increasing evidence suggests that also NASH patients with earlier fibrosis stages are at risk of disease progression, emphasizing the need for improved non-invasive risk stratification. Methods: We evaluated whether the apoptosis biomarker M30 can identify patients with fibrotic NASH despite low or intermediate FIB-4 values. Serum M30 levels were assessed by ELISA, and FIB-4 was calculated in an exploration (n = 103) and validation (n = 100) cohort of patients with histologically confirmed NAFLD. Results: The majority of patients with low FIB-4 (cut-off value < 1.3) in the exploration cohort revealed increased M30 levels (>200 U/L) and more than 80% of them had NASH, mostly with fibrosis. NASH was also detected in all patients with intermediate FIB-4 (1.3 to 2.67) and elevated M30, from which ~80% showed fibrosis. Importantly, in the absence of elevated M30, most patients with FIB-4 < 1.3 and NASH showed also no fibrosis. Similar results were obtained in the validation cohort. Conclusions: The combination of FIB-4 with M30 enables a more reliable identification of patients at risk for progressed NAFLD and might, therefore, improve patient stratification.

The relationship between oxidative stress and autophagy and apoptosis in patients with paroxysmal atrial fibrillation

Although the pathophysiology of paroxysmal atrial fibrillation (PAF) is not fully known, oxidative stress (OS) and atrial remodeling seem to be important triggers. Autophagy and apoptosis which are the types of cell death are fundamental processes in the human body. Although they investigated in many diseases, no study evaluated these parameters in PAF patients. We aimed to investigate autophagy and apoptosis which may be associated with atrial remodeling, and to show whether these factors are associated with OS in PAF patients. In this study, 44 PAF patients admitted to our clinic and 44 healthy volunteers were included. Serum total oxidative stress (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and ATG5 for autophagy and serum M30 for apoptosis were studied. Serum TOS, OSI, ATG5, M30 and left atrium (LA) diameter were higher, while TAS was lower in PAF group than the control group (p < 0.001, for all). ATG5 was positively correlated with TOS, OSI and LA, whereas negatively correlated with TAS. Also, M30 was positively correlated with TOS and OSI, whereas negatively correlated with TAS. Logistic regression analysis showed that TOS (P = 0.002), ATG5 (p = 0.013) and M30 (p = 0.006) were independent predictors of the PAF. It also found that ATG5 was the only independent predictor of LA enlargement in linear regression analysis. Our study showed that ATG5 and M30 were increased, and they were correlated with OS in patients with PAF. Therefore, we suggest that autophagy and apoptosis may play an important role in the PAF process.

Serum gasdermin D levels are associated with the chest computed tomography findings and severity of COVID-19.

BackgroundThe role of programmed cell death, especially pyroptosis and apoptosis, in unfavorable immune responses in COVID-19 remains to be elucidated.MethodsWe conducted a cross-sectional analysis to investigate the association between the serum gasdermin D (GSDMD) levels, a pyroptotic marker, and caspase-cleaved cytokeratin 18 fragment (M30), an apoptotic marker, and the clinical status and abnormal chest computed tomography (CT) findings in patients with COVID-19.ResultsIn this study, 46 patients diagnosed with COVID-19 were divided into the following three groups according to the disease severity: mild to moderate group (n = 10), severe group (n = 14), and critical group (n = 22). The serum GSDMD levels were higher in the critical group than in the mild to moderate group (P = 0.016). In contrast, serum M30 levels were lower in the critical group than in the severe group (P = 0.048). Patients who required mechanical ventilation or died had higher serum GSDMD levels than those who did not (P = 0.007). Area of consolidation only and of ground glass opacity plus consolidation positively correlated with serum GSDMD levels (r = 0.56, P < 0.001 and r = 0.53, P < 0.001, respectively).ConclusionHigher serum GSDMD levels are associated with critical respiratory status and the consolidation area on chest CT in patients with COVID-19, suggesting that excessive activation of pyroptosis may affect the clinical manifestations in patients with COVID-19.

Serum Cytokeratin 18 as a Metastatic and Therapeutic Marker for Extramammary Paget's Disease.

Extramammary Paget's disease (EMPD) is a rare cutaneous adenocarcinoma with unfavourable prognosis once it becomes invasive. A tumour marker that reflects disease progression is required for adequate management of EMPD. Cytokeratin 18 is highly expressed in many types of cancer and its soluble forms are detected by M30 (for caspase-cleaved form) and M65 (for both caspase-cleaved and intact forms) assays. We report here that tumour cells of EMPD in both lesional skin and lymph node metastasis are immunohistochemically positive for CK18, and the baseline serum M30 and M65 levels in patients with metastatic EMPD are significantly higher than those in non-metastatic patients. In addition, serial serum M30 and M65 levels might reflect recurrence of EMPD and response to chemotherapy. These results suggest that serum CK18 levels may be a useful tumour marker for advanced EMPD.

Relationship between hyperglycemia, insulin resistance and serum apoptosis marker m30- antigen in patients with type 2 diabetes mellitus

Objective: The amount of evidence suggests that the apoptosis marker M30-antigen (an antibody that recognizes the cytokeratin-18 fragment) has an association with hyperglycemia.&#x0D; Material and Methods: In this study, serum M30 levels of 145 patients diagnosed with prediabetes (n = 28) and type 2 diabetes, which was divided into four groups according to their HbA1c levels, were measured. The health control group (n = 24) was composed of healthy individuals. Serum concentrations of M30 antigen were measured using an enzyme-linked immunosorbent assay system and expressed as mean ± SD. HbA1c concentration was determined by boronate affinity technology according to NGSP standards.&#x0D; Results: M30 levels were comparable in the healthy control group (64,39±3,9 U/L) and prediabetes group (82,07±13,7). Type 2 diabetes Groups A, B, C, and D had levels of 109,38±16 U/L, 117,46±14,3 U/L, 173,69±48,1 U/L, and 163,40±37,3 U/L, respectively. The analysis of the data has shown that serum levels of M30 in the control and prediabetes groups were significantly lower than Type 2 diabetes Groups C and D (p=0.043). When all groups were taken into consideration, a significant relationship was found between HbA1c and serum M30 levels (r=0.231, p=0.002).&#x0D; Conclusion: Apparently, the increase in glycemia is followed by a rise in the serum levels of the, suggesting that apoptosis occurs as a secondary effect immediately after hyperglycemia.

Prognostic biomarkers in lung cancer patients in terms of long-term survival

Introduction:&#x0D; We aimed to investigate the predictive effect of serum M30 and M65 antigens on long-term prognosis in patients with advanced stage lung cancer before and after the first dose of chemotherapy.&#x0D; Methods:&#x0D; Fourty eight patients with advanced stage lung cancer were included in the study. Demographic data and histopathological characteristics of the patients were recorded. Serum levels of M30 and M65 were studied in 48 patients before chemotherapy, and in 43 patients both before and 48 hours after chemotherapy. Long-term survival was evaluated using Kaplan-Meier curves. The effect of high or low M30, M65 levels and M30/M65 ratio on long-term survival was investigated.&#x0D; Results: The mean age of the patients at the time of diagnosis was 57.52 ± 9.38 years. Fourty six of the 48 patients were men. While M30 value before chemotherapy was 163.23 ± 112.30 U/l; It was measured as 249.74 ± 266.67 U/l 48 hours after chemotherapy (p

Oxidative stress, DNA damage and apoptosis levels in those who use borderline high level fluoride content drinking water

Aim: Fluoride is necessary for tooth and bone development, but when exposed to excessive levels can cause oxidative stress, DNA damage, apoptosis, fluorosis and cancer. The aim of this study is to reveal the underlying mechanism of fluoride toxicity and to clarify, in part, the uncertainty of the fluoride level in the reference value ranges of drinking water. Material and Methods: Two groups were included in the study as exposure and control groups. Serum Total Oxidant Status and Total Antioxidant Status were measured with colorimetric; Urine 8-OHdG (8-hydroxy-2-deoxyguanosine) (CUSABIO) levels as DNA damage biomarkers and serum M30 and M65 levels (PEVIVA) as apoptosis biomarkers were studied by ELISA method. In addition, all participants underwent a dental examination by the dentist. Results: Serum total antioxidant status (TAS) were lower in the exposure group compared to the control group (p &amp;lt;0.001); serum total oxidant status (TOS) (p &amp;lt;0.001), OSI (p &amp;lt;0.001), M30 (p &amp;lt;0.001) and M65 (p &amp;lt;0.001) levels and urine 8-OHdG (p = 0.011) levels were high. However, the M30 / M60 ratio was not statistically different between the two groups (p = 0.371). Dental fluorosis was detected in all participants in the exposure group. Conclusion: This study showed increased levels of oxidative stress, DNA damage and apoptosis biomarkers in drinking water users with borderline high level fluoride. Therefore, instead of the World Health Organization's reference value range (0.5-1.5 mg/L) for fluoride levels in drinking water, the US Public Health Service’s (0.7 mg/L) reference value range seems to be more appropriate to the precaution.

Prediction of serum M30 and M65 levels for the short-term prognosis of patients with hepatitis B virus related acute-on-chronic liver failure during artificial liver support system therapy

Objective To analyze the dynamic changes of serum M30 and M65 levels in patients with hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) during artificial liver support system(ALSS) therapy, and to explore their predictive efficiency and clinical values for short-term prognosis of HBV-ACLF. Methods Seventy-six patients with HBV-ACLF who underwent ALSS therapy for the first time from May 2016 to May 2019 in the First Hospital of Jiaxing were selected.The patients were divided into improvement group (38 cases) and non-recovered group (38 cases)according to their prognosis, and 38 healthy persons were selected as control group during the same period.The serum levels of M30 and M65 were detected by enzyme-linked immunosorbent assay(ELISA). The predictive values of M30 and M65 levels for short-term prognosis in patients receiving ALSS were calculated by receiver operating characteristic analysis curve (ROC). M30 and M65 levels before and after ALSS were compared by two-way repeated measures analysis of variance. Results The levels of M30 and M65 in the improvement group, non-recovered group and control group were significantly different before the first ALSS therapy (F=109.36 and 90.42, respectively, both P 0.05). However, after twice ALSS therapy, the levels of M30 and M65 in non-recovered group were significantly higher than those in improvement group (t=30.699 and 64.777, respectively, both P<0.01). Moreover, after the second ALSS therapy, the levels of M30 and M65 were both significantly lower compared to those after the first-time therapy in the improvement group (t=3.350 and 5.932, respectively, both P<0.01). The areas under curve (AUC) of M30, M65 and the combination of M30 and M65 for prognosis prediction were 0.796, 0.844 and 0.906, respectively. The AUC of combination of M30 and M65 was significantly higher than M30 or M65 alone (Z=2.163 and 2.141, respectively, P=0.031 and 0.032, respectively). The cut-off values of M30 and M65 were 591.91 and 924.50 U/L, respectively. The sensitivity and specificity of combined M30 and M65 were 94.7% and 82.5%, respectively. Conclusions Serum M30 and M65 levels can predict the short-term prognosis of HBV-ACLF patients after ALSS therapy.The combination of M30 and M65 is of better diagnostic value. Key words: Liver, artificial; HBV-related acute-on-chronic liver failure; M30; M65

Multicenter Validation Study of a Diagnostic Algorithm to Detect NASH and Fibrosis in NAFLD Patients With Low NAFLD Fibrosis Score or Liver Stiffness

OBJECTIVES:Nonalcoholic steatohepatitis (NASH) and fibrosis play critical roles for the prognosis of patients with nonalcoholic fatty liver disease (NAFLD). Identification of patients at risk of NASH and fibrosis is therefore critical for disease management. NAFLD Fibrosis Score (NFS) and transient elastography (TE) have been suggested to exclude advanced fibrosis. However, there is increasing evidence that also patients with NASH and early fibrosis are at risk of disease progression and complications, emphasizing the need for improved noninvasive risk stratification in NAFLD.METHODS:Because hepatocyte apoptosis plays an early role in NASH pathogenesis, we evaluated whether the apoptosis biomarker M30 might identify NAFLD patients who are at risk of NASH and fibrosis despite low NFS or TE values. Serum M30 levels were assessed by enzyme-linked immunosorbent assay in combination with NFS and/or TE in an exploration (n = 103) and validation (n = 100) cohort of patients with biopsy-proven NAFLD.RESULTS:Most patients with low NFS (cutoff value < −1.455) revealed increased M30 levels (>200 U/L) in the exploration (62%) and validation (67%) cohort, and more than 70% of them had NASH, mostly with histological fibrosis. Vice versa, most patients with NFS < −1.455 but nonelevated M30 levels showed no NASH. NASH was also detected in most patients with indeterminate NFS (−1.455 to 0.676) but elevated M30 levels, from which ∼90% showed fibrosis. Similar results were obtained when using TE instead of NFS.DISCUSSION:The combination of the M30 biomarker with NFS or TE enables a more reliable identification of patients with an increased risk of progressed NAFLD and improves patient stratification.

Usefulness of serum M30 and M65 levels to predict response to neoadjuvant chemotherapy in patients with breast cancer

ObjectiveM30 and M65 levels reflect tumor cell activity in patients with epithelial cancer. Cytokeratin 18 is one of the cell skeletal elements. M30 is a apoptotic marker of cytokeratin 18. M65 levels are both an apoptosis and a necrosis marker. The aim of our study was to determine the predictive value of M30 and M65 levels in neoadjuvant treatment of breast cancer. Materials and methodsIn this prospective study, 41 patients with breast cancer who underwent neoadjuvant chemotherapy were included. Following 4 cycles of chemotherapy with anthracycline containing regimen, patients received paclitaxel treatment for 12 weeks. Blood was collected from the patients before chemotherapy and on day 21, after the 2nd, 4th, and 8th cycles. M30 and M65 levels were measured with the ELISA method. ResultsWhile there was an increase in M30 and M65 levels at the 4th cycle (P < 0.05), levels were decreased after the 8th cycle. In addition, there was no significant relationship among M30, M65 levels, and prognostic factors such as ER, PR, c-Erb-2, Ki-67, pathologic-T, pathologic-N, and chemotherapy responses. ConclusionM30 and M65 levels are not of predictive values of response to breast cancer patients receiving neoadjuvant chemotherapy. Nevertheless, M30 and M65 levels increased when patients kept receiving anthracycline containing chemotherapy.

Evaluation of serum M30 and M65 activity in patients with stage-I endometrial cancer

We aimed to analyse the prognostic value of serum oxidative stress parameters and apoptotic markers of serum M30/65 levels in endometrial cancer patients. Serum M30/65 levels and oxidative stress parameters were evaluated in 52 women with stage I endometrial cancer (n = 26) and a control group of healthy females (n = 26). The total antioxidant status (p = .002), oxidative stress index (p = .003) and serum M30/65 levels (p < .001) were significantly higher in women with stage-I endometrial cancer in comparison to the control group. Furthermore, serum M30/65 levels were significantly lower on postoperative day 8, compared to preoperative levels (p = .001 and p < .001, respectively), in the endometrial cancer group. Although impaired apoptotic activity plays a crucial role in the aetiopathogenesis of endometrial cancer, oxidative stress may be instrumental in malignant transformation. We concluded that measurement of M30/65 levels would be beneficial in the follow-up of women with endometrial cancer.Impact StatementWhat is already known on this subject: Although M30 has been evaluated as a marker of apoptosis in tissue samples from women with endometrial cancer (EC), no previous studies have simultaneously analysed serum M30 and M65 levels and oxidative stress in patients with stage-I EC.What the results of this study add: Total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI) and serum M30/65 levels were significantly higher in women with stage I EC in comparison to the control group. Furthermore, serum M30/65 levels were significantly lower on postoperative day 8, compared to preoperative levels, in the EC group. The fact that pre-operative M30/M65 levels were higher than the post-operative levels may be very important in early-stage ECWhat the implications are of these findings for clinical practice and/or further research: Although impaired apoptotic activity plays a crucial role in the aetiopathogenesis of EC, oxidative stress may be instrumental in malignant transformation. The fact that serum M30/M65 levels decreased in accordance with the reduction of post-operative tumour burden led us to conclude that measurement of M30/65 levels would be beneficial in the follow-up of women with EC.

Hepatic sonic hedgehog protein expression measured by computer assisted morphometry significantly correlates with features of non-alcoholic steatohepatitis

BackgroundHepatic expression of Sonic Hedgehog (SHH) is associated with Non-alcoholic fatty liver disease (NAFLD) and development of Non-alcoholic steatohepatitis (NASH). Hepatic SHH detection increases with the diagnosis of NASH. This pilot study was designed to confirm that staining for SHH is useful in NASH diagnosis and determine whether quantification of staining by computer assisted morphometry (CAM) can be used to assess severity of ballooning degeneration.MethodsSHH was detected by immunohistochemistry (IHC) on paraffin-embedded liver sections in subjects (N = 69) with biopsy proven NAFLD and no liver disease (control). Serum samples were also available for these subjects. Post-staining, a digitized image of the section was acquired and an area quantification algorithm was used to quantify the degree of SHH expression. Additionally, circulating M30, M65, and SHH were measured by ELISA.ResultsNotably, hepatic SHH expression correlated with histologic ballooning degeneration (rho = 0.62, p < 0.0001), steatosis grade (rho = 0.554, P < 0.001), Mallory-Denk bodies (rho = 0.54, P < 0.001), pericellular fibrosis (rho = 0.527, P < 0.001), and lymphocytic infiltration (rho = 0.435, P < 0.0002). Additionally, hepatic SHH expression correlated with circulating M65 (rho = 0.588, p < 0.0001), and circulating M30 (rho = 0.375, p = 0.001), as well as AST and ALT (rho = 0.43, p = 0.0004, and rho = 0.27, p = 0.03, respectively). Further, serum M30 was almost twice as high in NASH patients compared to non-NASH (539.1 ± 290.8 U/L vs. 287.6 ± 190.5 U/L; p = 0.0002), while M65 was almost three times higher in NASH patients compared to non-NASH (441.2 ± 464.2 U/L vs. 162.8 ± 353.1 U/L, P = 0.0006). Logistic modeling indicates hepatic SHH expression and presence of type 2 diabetes as independent predictors of advanced fibrosis (defined as portal and pericellular fibrosis > 2: OR = 1.986, p = 0.01, and OR = 3.280, p = 0.03, respectively).ConclusionThus, our findings show quantitation of SHH expression by CAM can provide a tool for quantifying changes in hepatocyte injury and assist in unambiguous staging/grading of NASH. Our study showed minimal interobserver variability using CAM based quantification. Once validated, CAM assessment of hepatic SHH could benefit clinical trials or long term outcomes studies of NASH subjects.

The roles of M30 and M65 in the assessment of treatment response and prognosis in patients with non-small cell lung cancer, who receive neoadjuvant treatment.

Aim of the studyTo investigate the efficacy of evaluating prognosis and response to lung cancer treatment using M30 and M65 antigens, which are indicators of necrosis.Material and methodsForty-eightpatients with lung cancer, who were planned to receive neoadjuvant chemotherapy, and 38 healthy volunteers were enrolled in the study. Using M30 and M65 levels, cytokeratin 18 levels were measured twice: before and 48 hours after the first chemotherapy treatment. Apoptotic and total necrosis levels were determined by measuring the M65 and M30 levels.ResultsThe M30 and M65 antigen levels in the patient group were significantly higher than in the control group (p< 0.001). The M30 and M65 antigen levels were significantly higher 48 hours after the chemotherapy compared with before the chemotherapy (p< 0.001). There were no significant differences in M65 levels between patients who responded to treatment and patients who progressed. The M30 levels increased significantly in patients with disease progression (p= 0.694 and p = 0.024, respectively). No significant differences in serum M30 and M65 antigen levels were found when compared between the surviving and deceased patients (p = 0.126 and p = 0.340, respectively).ConclusionsA significant increase was detected in serum M30 and M65 levels in patients with lung cancer. There was a greater increase in serum M30 levels in patients who did not respond to the chemotherapy. This result gives rise to the thought that evaluating apoptosis and total necrosis through M30 and M65 measurements alone only in patients receiving neoadjuvant chemotherapy would be insufficient for specifying the effectiveness of the treatment.

Serum Caspase-Cleaved Cytokeratin (M30) Indicates Severity of Liver Dysfunction and Predicts Liver Outcome.

BackgroundThe Model for End-Stage Liver Disease (MELD) score is a well-established tool for assessing hepatic failure. The present retrospective study investigated whether serum keratin 18 (M65) and caspase-cleaved cytokeratin (M30) were associated with liver dysfunction and post-transplant graft failure.Material/MethodsA total of 147 patients with liver cirrhosis were categorized into 2 groups according to their baseline MELD score (group I: MELD score <20, n=87, and group II: MELD score ≥20, n=60). Serum M65 and M30 levels were measured by ELISA.ResultsCirrhotic patients had significantly higher serum M65 and M30 levels than healthy controls (p<0.0001). Serum M65 was correlated with the MELD score and serum bilirubin (p≤0.007) and serum M30 was correlated with the MELD score, international normalized ratio, and serum bilirubin (p≤0.001). Group II had significantly higher serum M65 and M30 levels than group I (M65, p=0.025 and M30, p<0.001). Patients who lost the allograft during the first post-transplant year had significantly higher serum M30 levels than patients with a graft survival of >1 year (p=0.004). In the regression analysis, serum M30 was associated with the MELD score (odds ratio [OR]=2.545, p=0.005), serum bilirubin (OR=2.605, p=0.005) and 1-year graft loss (OR=3.61, p=0.006).ConclusionsOur data indicate that serum M30 levels reflect the degree of liver dysfunction and can predict 1-year graft loss.

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.