M30 Levels Research Articles

Can circulating M30 and M65 levels be beneficial markers in the diagnosis and management of patients with complete hydatidiform mole?

The objective of this study is to evaluate the prognostic value of M30 and M65 levels as markers of apoptotic activity and maternal serum oxidative stress in patients with complete hydatidiform mole (CHM). In total, 68 pregnant women were included in the study. The study group included 34 pregnant with CHM, while 34 healthy pregnant were employed as a control group. Venous blood samples were drawn to assess the maternal serum oxidative stress and M30-M65 levels. In addition, a second blood sample was drawn from patients with CHM on day 8 after dilatation evacuation. Maternal serum oxidative stress and M30-M65 levels were found to be significantly higher in patients with CHM as compared with the control group. It was found that serum β-subunit of human chorionic gonadotropin (β-hCG) level had a significant positive correlation with M30-M65 levels in patients with CHM. In addition, serum M65 level was found to be as effective as β-hCG in the identification of the patients with CHM. Our results indicated that oxidative stress and apoptosis may play significant roles in CHM development. In addition, it seems that serum M30-M65 levels can presumably be an ancillary laboratory test to β-hCG in the diagnosis and follow-up of the patients with CHM.

The M30 assay does not detect apoptosis in epithelial-derived cancer cells expressing low levels of cytokeratin 18

The primary aim of this study was to compare measurement of apoptosis by M30 immunoreactivity (a biomarker for apoptosis) to other apoptosis assays (morphological assessment of nuclei, Annexin-V-FITC staining, DNA fragmentation and PARP cleavage) in vitro. Caspase-cleaved cytokeratin 18 (M30, ccK18) is only produced in epithelial cells and is regarded as a pharmacodynamic biomarker of apoptotic cell death because it is released from cells during apoptosis induced by chemotherapeutic agents. However, we have observed false negative results using this assay in clinical samples. Therefore, we tested its ability to accurately detect apoptosis in a panel of lung cancer cell lines with a range of clinically approved chemotherapeutic drugs. Three different non-small cell lung cancer (NSCLC) cell lines (A549, H1299, PC3) were used to correlate M30 levels with alternate apoptosis assays. Following successful induction of apoptosis, the A549 cell line showed an increase in M30 levels along with other well-known features of apoptosis, whilst H1299 and PC3 cell lines did not show an increase in M30 levels, even when apoptosis was detected by other means. Further analysis showed that H1299 and PC3 cell lines expressed much lower levels of cytokeratin 18 protein compared to the A549 cell line. Our results suggest that reliable detection of apoptosis via the M30 assay only works when sufficient levels of cytokeratin 18 are present in the cells. This means that the M30 assay may result in false negative results for apoptosis, and as such, the ELISA should be used in conjunction with other assays.

(247) - Ex-Vivo Lung Perfusate Cell Death Markers May Predict Long Term Outcomes After Transplantation

Purpose: Ex-Vivo Lung Perfusion (EVLP) is an emerging technology to reas- sess marginal donor lungs for transplantation. We have previously reported that cell death related perfusate bio-markers in EVLP are potentially predic- tive for primary graft dysfunction (PGD). Our objective was to investigate whether EVLP perfusate cell death markers could be used to predict post- transplant long term survival. Methods: Caspase-cleaved cytokeratin 18 (M30) and HMGB-1 levels in the perfusate of EVLP of marginal donor lungs were measured by quantitative enzyme linked immunosorbent assay, and were correlated with short and long term clinical outcome. Results: From 100 sequential clinical EVLP cases, 79 lungs were selected to be transplanted. Recipients that were bridged with pre-transplant extra- corporeal life support system were excluded from this analysis. 11 patients (PGD group) developed PGD 3 at any time within 72 h after transplant following bilateral lung transplant (BLTx). 34 patients (Control group) did not develop PGD grade 3 after BLTx. Levels of M30 were signifi- cantly higher in PGD group compared to Control group at 1 h (Control 53.9±15.9 vs. PGD 73.3± 24.9 U/L, p= 0.004) and 4 h (Control 72.4±40.0 vs. PGD 137.0±146.6 U/L; p= 0.04) of EVLP . The increase of HMGB-1 from 1 h to 4 h of EVLP (delta HMGB-1) in PGD group was signifi- cantly higher than Control (Control 7.2±16.8 vs. PGD 37.0±25.4 ng/ml; p= 0.0002). Uni variate Cox regression analysis revealed that higher M30 at 1 h (HR= 1 .03 (95%CI 1.004-1.058) per 1 U/L; p= 0 .025) and 4 h (HR= 1.009 (95%CI 1.002-1.016) per 1 U/L; p= 0.009), and higher delta HMGB-1 (HR= 1.033 (95%CI 1.006-1.061) per 1 ng/ml; p= 0.016) are sig- nificantly predictive for survival. After adjustment for PGD using bivariate cox regression analyses, M30 at 4h remained significant (p= 0.04) and the other markers were not statistically significant (M30 at 1 h; p= 0.13; delta HMGB-1; p= 0 .10). The significance of M30 at 4 h seen after adjustment for PGD may suggest that the marker provides additional predictive power for survival above that predicted by PGD status alone. Conclusion: We have demonstrated a correlation of cell death markers in the EVLP perfusate with PGD outcome and long term survival. Further research and validation is required to determine which of these biomarkers will be important in repertoire for real time predictive evaluation of the ex vivo perfused donor lung.

P1296 : Blocking NMDA receptors prolongs survival in rats with acute liver failure by dual protective mechanisms in kidney and brain

were 1094.67±239.37mIU/ml. The difference in values between these groups was statistically significant (p = 0.004). There was a statistically significant difference in total leucocyte count, serum creatinine, INR and cleaved CK18 M30 levels among those that survived vs expired. Multivariate analysis found that all these independently predict mortality. A receiver operating characteristic curve was drawn for the new score. The score found was compared with the King’s College criteria and the MELD score, the new scoring system was found to have higher sensitivity and specificity for mortality compared with other standard prognostic markers in ALF, and has a better diagnostic accuracy (Table 1).

Maternal and fetal serum levels of caspase-cleaved fragments of cytokeratin-18 in intrahepatic cholestasis of pregnancy

Objective: We aimed to investigate the relationship between intrahepatic cholestasis of pregnancy (ICP) and caspase-cleaved fragments of cytokeratin-18, also referred to as M30, a marker of apoptosis. Methods: In this case–control study, maternal and umbilical cord blood venous samples were obtained from 21 pregnant women with ICP and 22 healthy pregnant women as a control group. M30 levels were compared among the groups. Results: Maternal serum M30 levels were significantly higher in the severe ICP group than in the control (p < 0.001) and mild ICP groups (p = 0.006). The values were comparable between the mild ICP and the control groups. The umbilical cord serum M30 levels were also significantly greater in the severe ICP group than in the control group (p = 0.001). Conclusions: Changes in M30 levels, as an apoptosis marker, may shed light on the pathogenesis of ICP. Explaining the mechanisms of bile acid (BA)-induced hepatocyte injury may contribute further therapeutic strategies for the treatment of human cholestatic diseases.

Utility of M30, an apoptotic serum marker, in liver diseases.

The aim of this paper is to evaluate the role of apoptosis in some common liver diseases, and the utility of M30, an apoptotic serum marker, in the diagnosis of the severity of underlying hepatic injury. As is widely known, apoptosis is programmed cell death, and its deregulation results in an uncontrolled inflammatory process leading to upregulation of liver fibrogenesis. Both extrinsic and intrinsic pathways are crucial in apoptosis, and caspase cleavage of cytokeratin proteins occurs in both. Therefore, the measurement of caspase- cleaved cytokeratin fragments could be a novel method to assess the intensity of apoptotic cell numbers in epithelial tissue damage. M30 levels were found to increase not only in acute liver disorders, but also in some chronic liver injuries. We tried to summarize the recent studies focused on the role of apoptotic processes in liver diseases, mainly those that investigated the use of M30 in determining the severity of, or in predicting, ongoing liver injury.

Mini-Laparoscopy Guided Liver Biopsy Increases Diagnostic Accuracy in Acute Liver Failure

Background/Aims: For diagnosis, prognosis, and treatment of acute liver failure (ALF), macroscopic evaluation and histological assessment of the liver are important. Due to impaired coagulation in ALF, the risk of bleeding is high after a percutaneous liver biopsy. Our aims were to assess (i) safety and benefit of mini laparoscopy (ML) in patients with ALF and (ii) the potential utility of histological markers in ALF prognosis. Methods: ML was performed in 39 patients with ALF to assess liver surface and to obtain a liver biopsy. Serological markers of liver injury and immunohistochemical detection of cell death and proliferation were compared to a non-ALF group (n = 10). Results: Liver biopsies were successfully performed in all patients with no significant complications. All patients had markedly elevated M30 and M65 levels in the serum. In the liver, M30 and Ki67 immune-reactive cells were more abundant in those with ALF. Importantly, there were significantly more Ki67-positive cells but fewer M30-positive cells in livers of ALF patients who recovered spontaneously. Conclusion: ML with liver biopsy in patients with ALF and severe coagulopathy is safe. Immunohistochemical detection of liver cell death and regeneration may identify individuals who would recover spontaneously or who would need a liver transplant.

Effects of off-pump versus on-pump coronary artery bypass grafting: apoptosis, inflammation, and oxidative stress.

It has been suggested that off-pump coronary artery bypass grafting (CABG) surgery reduces myocardial ischemia-reperfusion injury, postoperative systemic inflammatory response, and oxidative stress. The aim of this study was to measure serum malondialdehyde (MDA), high-sensitivity C-reactive protein (hs-CRP), M30, and M65 levels and to investigate the relationship between M30 levels and oxidative stress and inflammation in patients undergoing on-and off-pump CABG surgery. Fifty patients were randomly assigned to on-pump or off-pump CABG surgery (25 patients off-pump and 25 on-pump CABG surgery), and blood samples were collected prior to surgery, and 30 minutes, 60 minutes, 6 hours, and 24 hours after CABG surgery. Compared to the on-pump group, serum MDA levels at 30 minutes, 60 minutes, 6 hours, and 24 hours after the CABG surgery were significantly lower in the off-pump group (P=.001, P=.001, P=.001, and P=.001, respectively). Serum M30 levels were found to be elevated in both groups, returning to baseline at 24 hours. When compared to baseline, the hs-CRP level reached its peak at 24 hours at 13.28±5.32 mg/dL in the on-pump group, and 15.44±4.02 mg/dL in the off-pump group. CABG surgery is associated with an increase in inflammatory markers and serum M30 levels, indicating epithelial/endothelial apoptosis in the early period.

Effective inhibition of cardiomyocyte apoptosis through the combination of trimetazidine and N-acetylcysteine in a rat model of myocardial ischemia and reperfusion injury

Objective: Apoptosis is the early and predominant form of cell death in infarcted myocardia. The aim of the study was to investigate the effects of trimetazidine (TMZ) and N-acetylcysteine (NAC), used alone or in combination, on oxidative stress, infarct size, and ischemia-reperfusion (IR)-induced cardiomyocyte apoptosis in a rat model of myocardial IR. Methods and results: Myocardial IR was established by ligating an area under the left main coronary artery for 30 min followed by 3 h of reperfusion. Saline (1 ml/kg), NAC (50, 150 mg/kg), or TMZ (3, 5 mg/kg) was intravenously injected during the middle of the ischemic period. At the end of the reperfusion, blood samples were collected from the animals to measure serum M30 and M65 levels, which are markers of cell death, the S100b level, which is a marker of inflammation, and the malondialdehyde (MDA) level, which is a marker of oxidative stress. The infarct size was evaluated as the ratio of the infarct area to the risk area. Apoptotic activation was assessed by caspase-3 immunostaining and a TUNEL assay. TMZ and NAC, either alone or in combination, significantly reduced serum MDA levels, infarct area and apoptotic activity compared to those observed in saline group. Interestingly, the infarct area was more smaller in TMZ (3 and 5 mg/kg) injected groups (9.72 ± 1.3% and 9.96 ± 2.3%) than those observed in NAC (50 and 150 mg/kg) (16.1 ± 2.5% and 19.1 ± 2.14%) or TMZ (5 mg/kg)- NAC (150 mg/kg) combination groups (16.9 ± 1.6%). However, the apoptotic activity was reduced more significantly in the combination of TMZ (5 mg/kg)-NAC (50 mg/kg) compared to TMZ-only group. Neither TMZ or NAC treatments nor the combination of the drugs significantly affected serum M30, M65 and S100B levels. Conclusion: Intravenous NAC and TMZ administration decreased oxidative stress, infarct area and apoptotic activity in a rat model of IR. Although the combination treatment was more effective in reducing the apoptotic activity than either treatment groups alone, TMZ treatment was more successful in reducing the infarct area than NAC or combination treatments. Present results suggest that, in addition to mechanical attempts to secure myocardial reperfusion, the use of TMZ and NAC may help to reduce IR injury.

Clinical significance of serum M30 and M65 levels in patients with breast cancer

M30 and M65 are relatively new assays that detect different circulating forms of the epithelial cell structural protein cytokeratin18. The objective of this study was to determine the clinical significance of the serum levels of M30 and M65 in patients with breast cancer. A total of 80 patients with a pathologically confirmed diagnosis of breast cancer were enrolled into the study. Serum M30 and M65 concentrations were determined by the solid-phase sandwich ELISA method. Serum samples were obtained on first admission before any type of treatment. The median age at diagnosis was 52 years, range 30 to 81 years. The baseline serum M30 and M65 levels in patients with metastatic disease were significantly higher than those in the non-metastatic patients (P=0.017 and P=0.003, respectively). Moreover, serum M65 level was also elevated in patients with large tumor size (P=0.02). No correlation was found between these serum assay levels and response to chemotherapy (P>0.05). However, the significant relationship was found between the serum levels of M30 and M65 (rs=0.96, P<0.001). Neither serum M30 nor serum M65 had significantly effect on survival (P=0.50, and P=0.52, respectively). In conclusion, although both serum M30 and M65 levels are elevated in metastatic disease, no predictive and prognostic roles on survival were found in patients with breast cancer.

Circulating biomarkers in hepatocellular carcinoma

Our aims are to determine levels of circulating cellular and protein biomarkers in hepatocellular carcinoma (HCC) patients and to analyse any relationships with clinical parameters. Fifty-four consenting patients were recruited. Circulating tumour cells (CTCs) were enumerated (by CellSearch) and characterised via filtration [by isolation by size of epithelial tumour cells (ISET)] with downstream immunohistochemistry (IHC). Glypican-3 (GPC3) expression in tumour biopsies and CTCs (by IHC) was compared, and levels of circulating caspase-cleaved and full-length cytokeratin 18 (CK18, measured using M30 and M65 ELISAs) were examined as a putative prognostic factor and marker of tumour burden. CTCs were identified in 14 out of 50 (28%) patients by CellSearch and in 19 out of 19 (100%) patients by ISET. The presence of GPC3-positive CTCs by ISET was 100% concordant with the presence of GPC3-positive cells in the original tumour (n = 5). No statistically significant correlations were observed between CTC number and clinical characteristics, although trends were noted between CTC subtypes, Child-Pugh score and tumour node metastasis stage. Serum M30 and M65 levels (as continuous variables) significantly correlated with overall survival (OS) in a univariate analysis (p = 0.003 and p < 0.001, respectively); M65 levels remained statistically significant in a multivariate analysis (p = 0.029). This is the first study to detect GPC3-positive CTCs in HCC, important for drug development with this target. The significant association of circulating CK18 with OS in HCC further exemplifies the utility of circulating biomarkers in cancer.

PTH-082 Do Serum Markers Of Cell Injury And Death Have Potential To Become Mechanistic Markers In Non-alcoholic Fatty Liver Disease (nafld)?

IntroductionDegree of hepatocellular injury, necrosis/apoptosis and inflammation may be assessed by the serum markers that reflect the pathogenic process. We investigated the correlation of circulating miR-122, High Mobility Group Box-1...

P485 THE RATIO OF ASCITES CALPROTECTIN TO TOTAL PROTEIN IS A DIAGNOSTIC AND PROGNOSTIC MARKER FOR SPONTANEOUS BACTERIAL PERITONITIS IN LIVER CIRRHOSIS

inclusion. The median follow up time was 283 days (range 1– 1382 days). M30 levels differed between Child Pugh stages and correlated with the MELD score (model of end stage liver disease). When patients were grouped according to M30 levels in individuals with low ( 3000U/l) M30 levels, M30 could differentiate overall survival of patients according to the log-rank test (P < 0.001). Furthermore, M30 level was an independent risk factor for mortality in addition to age, gender, Child Pugh stage and the MELD score in a multivariate Cox regression model. Conclusions: The M30 serum level correlates with the stage of cirrhosis and is predictive for survival in patients with liver cirrhosis.

M30 Does Not Predict the Severity of Hepatosteatosis, Whereas Adiponectin Level Declined With Increase of ALT and the Severity of Hepatic Steatosis

Nonalcoholic fatty liver disease (NAFLD) is an emerging problem all over the world. Because NAFLD and polycystic ovary syndrome (PCOS) are both closely related with insulin resistance, it would be necessary to determine the rate of presence of NAFLD in PCOS patients. So, this study aimed to investigate the utility of M30 in PCOS patients for the diagnosis of hepatic injury. Eighty patients with PCOS were included in the study. Ultrasonographic examination for the presence of hepatic steatosis, M30 serum level for determining the severity of ongoing apoptotic cell death in liver, and BARD index for defining the hepatic injury were performed during the study. 25-OH vitamin D and adiponectin level in sera were studied using ELISA (Enzyme-Linked ImmunoSorbent Assay). M30 and vitamin D levels did not change significantly with the severity of hepatic steatosis. On the other hand, M30 levels showed a positive correlation with ALT and AST levels, and M30 level suddenly increased with the presence of hepatic steatosis from 159.7 to 170 U/l, however stabilized with the increasing severity of hepatic setatosis. Adiponectin levels decreased with the increasing severity of hepatic steatosis and significantly varied between ALT greater than 40 U/l and less than 40 U/l. M30 level in serum increased with the appearance of hepatic steatosis and had a positive correlation with a noninvasive hepatic injury test, BARD (BMI, aspartate aminotransferase [AST]/alanine aminotransferase [ALT] ratio [AAR], diabetes mellitus [DM]) index. Adiponectin level decreased with the increasing ALT level and severity of hepatic steatosis.

Soluble intracellular adhesion molecule, M30 and M65 as serum markers of disease activity and prognosis in cholestatic liver diseases

Hepatic apoptosis is involved in the pathogenesis of immune-mediated liver diseases such as autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). The aim of our study was to quantify distinct markers of apoptosis in sera of patients with AIH, PBC and PSC, and to evaluate correlation with markers of disease activity and prognosis. Sera of patients with AIH, PBC and PSC, and of healthy controls were collected and distinct cell death markers were quantified using a bead-based multiplex enzyme linked immunosorbent assay (soluble intracellular adhesion molecule [sICAM], macrophage migration inhibitory factor [MIF], soluble Fas [sFas], plasminogen activator inhibitor 1 [PAI-1]) or single enzyme-linked immunosorbent assays (DNAse, M30, M65). In comparison with healthy controls, the apoptotic markers sFas, sICAM (only in PSC patients), M30 and the cell death marker M65 were substantially elevated in sera of patients with immune-mediated liver diseases, whereas DNAse activity was reduced. Interestingly, patients with advanced PSC presented with higher levels of sICAM, M30 and M65 than patients with mild PSC. Regression analysis revealed correlations between serum levels of sICAM, M30 and M65 with the Mayo Risk Score for PSC, and of M65 with the Mayo Risk Score for PBC. Concentrations of the serum markers of apoptosis sFas and M30 and of the marker of total cell death M65 are elevated in patients with immune-mediated liver diseases, whereas activity of DNAse is reduced. In patients with PSC, sICAM, M30 and M65 may serve as indicators for disease activity and prognosis.

Use of monitoring levels of soluble forms of cytokeratin 18 in the urine of patients with superficial bladder cancer following intravesical Ad-IFNα/Syn3 treatment in a phase l study

A phase l study using intravesical Ad-IFNα/Syn3 for patients with BCG resistant superficial bladder cancer showed a complete remission (CR) of 43% at 90 days after treatment with high levels of IFNα being produced. Ad-IFNα kills bladder cancer cells by two apoptotic and one necrotic mechanism that can be measured by soluble forms of cytokeratin 18 (CK 18) using M30 and M65 ELISAs, assays for caspase –cleaved (apoptotic) and uncleaved (necrotic) cell death, respectively. Therefore we determined whether M30 and M65 levels in the urine after treatment could document all three mechanisms of cancer cell kill and also predict having a CR. High levels of both M30 and M65 were found in all patients within 24 hours after treatment with all three types of cancer cell death occuring. Moreover, the return of both M30 and M65 levels in the urine to normal levels within 5 days or more after treatment was strongly associated with obtaining a CR (p=0.003). This is the first time that such assays have been used to study response to therapy in the urine of patients with bladder cancer and in the future may prove valuable in predicting clinical outcome.

Pure anti-tumor effect of zoledronic acid in naïve bone-only metastatic and locally advanced breast cancer: proof from the “biological window therapy”

The study investigated the anti-tumour effect of zoledronic acid (ZA) administered alone in a biological window therapy in naïve bone-only metastatic and locally advanced breast cancer (LABC) patients. 33 patients with LABC (Group 1) and 20 patients with a first diagnosis of bone metastasis only (Group 2) received 4 mg single dose of ZA, 14 days (biological window) before starting any treatment. In Group 1, Ki67, CD34, p53/bcl-2 and caspase 3 expression along with the adenosine triphosphate (ATP) levels and RNA disruption index were evaluated as markers of tumor growth in tumour specimens obtained before and after ZA administration (basal, day 14). In Group 2, the total enumeration of circulating tumour cells (CTCs), and of M30+ve CTCs along with the soluble marker of cell death (M30/M65) were carried-out as markers of tumor dissemination at baseline, at 48 h and day 14th. In Group 1, there was a significant reduction in Ki67, CD34, bcl-2 expression after 14 days ZA based-treatment (p = 0.0032; p = 0.0001, p < 0.00001 respectively). ZA showed a significant increase of RNA disruption (p < 0.0076). In Group 2, we observed a significant reduction of CTCs number after 48 h (p = 0.0012), followed by a significant rebound at 14 days (p = 0.012). The apoptotic CTCs/M30+ve and M65 levels significantly increased under treatment (p = 0.018 and p = 0.039 respectively) after drug administration when compared to the baseline. These results are the first prospective in vivo data showing the direct pure anti-tumour effect (either on the tumour cell or on CTCs) of ZA.

Serum CK18 as a Predictive Factor of Response to Chemotherapy in Locally Advanced and Metastatic Breast Cancer

Introduction: Breast cancer is the most common cancer in women and the second most frequent cause of cancer death. Several factors affect response to chemotherapy including nodal status, hormonal status and human epidermal growth factor receptor (Her-2). Aim of Study: The study is aiming at evaluating M30 antigen in serum of patients with locally advanced and metastatic breast cancer and establishing the relation between M30 level and response to chemotherapy. Patients and Methods: The study was performed at Al Bairouni University Hospital and the Faculty of Pharmacy (Damascus-Syria). We have included 60 patients with histologic confirmation of invasive ductal carcinoma of the breast treated with the combination (Docetaxel + Doxorubicin) with M30 levels to be evaluated before treatment and 24 hours after the first and third cycle. Results: M30 level increase in serum 24 hours after the 1st cycle correlated with different kinds of response in 39 patients (P value less than 0.03) with better results in those with Estrogen Receptors (ER) positive patients (P value 0.05). There was no correlation between Her-2 status and response (P value 0.3). Conclusion: M30 level in serum is a useful predictor marker of response to chemotherapy in both locally advanced and metastatic breast cancer.

The Significance of Caspase-Cleaved Cytokeratin 18 in Pleural Effusion

BackgroundApoptosis plays a role in the development of pleural effusion. Caspase-cleaved cytokeratin 18, a marker for epithelial cell apoptosis, was evaluated in pleural effusion.MethodsA total of 79 patients with pleural effusion were enrolled. The underlying causes were lung cancer (n=24), parapneumonic effusion (n=15), tuberculous effusion (n=28), and transudates (n=12). The levels of M30, an epitope of caspase-cleaved cytokeratin 18, were measured in blood and pleural fluids using enzyme-linked immunosorbent assay along with routine cellular and biochemical parameters. The expression of M30 was evaluated in the pleural tissues using immunohistochemistry for M30.ResultsThe M30 levels in pleural fluid were significantly higher in patients with tuberculosis (2,632.1±1,467.3 U/mL) than in patients with lung cancer (956.5±618.5 U/mL), parapneumonic effusion (689.9±413.6 U/mL), and transudates (273.6±144.5 U/mL; all p<0.01). The serum levels were not significantly different among the disease groups. Based on receiver operating characteristics analysis, the area under the curve of M30 for differentiating tuberculous pleural effusion from all other effusions was 0.93. In the immunohistochemical analysis of M30, all pathologic types of cancer cells showed moderate to high expression, and the epithelioid cells in granulomas showed high expression in tuberculous pleural tissues.ConclusionCaspase-cleaved cytokeratin 18 was most prominently observed in tuberculous pleural effusion and showed utility as a clinical marker. The main source of M30 was found to be the epithelioid cells of granulomas in tuberculous pleural tissues.

Non-invasive diagnosis of alcoholic liver disease.

Alcoholic liver disease (ALD) is the most common liver disease in the Western world. For many reasons, it is underestimated and underdiagnosed. An early diagnosis is absolutely essential since it (1) helps to identify patients at genetic risk for ALD; (2) can trigger efficient abstinence namely in non-addicted patients; and (3) initiate screening programs to prevent life-threatening complications such as bleeding from varices, spontaneous bacterial peritonitis or hepatocellular cancer. The two major end points of ALD are alcoholic liver cirrhosis and the rare and clinically-defined alcoholic hepatitis (AH). The prediction and early diagnosis of both entities is still insufficiently solved and usually relies on a combination of laboratory, clinical and imaging findings. It is not widely conceived that conventional screening tools for ALD such as ultrasound imaging or routine laboratory testing can easily overlook ca. 40% of manifest alcoholic liver cirrhosis. Non-invasive methods such as transient elastography (Fibroscan), acoustic radiation force impulse imaging or shear wave elastography have significantly improved the early diagnosis of alcoholic cirrhosis. Present algorithms allow either the exclusion or the exact definition of advanced fibrosis stages in ca. 95% of patients. The correct interpretation of liver stiffness requires a timely abdominal ultrasound and actual transaminase levels. Other non-invasive methods such as controlled attenuation parameter, serum levels of M30 or M65, susceptometry or breath tests are under current evaluation to assess the degree of steatosis, apoptosis and iron overload in these patients. Liver biopsy still remains an important option to rule out comorbidities and to confirm the prognosis namely for patients with AH.