Andrzej Gabriel

Chemerin, vaspin and insulin resistance in chronic hepatitis C

Summary.  Adipocytokine profile seems to play a distinct role in the pathogenesis of chronic hepatitis C (CHC). Chemerin and vaspin are recently described adipocytokines with various suggested functions and potential to modulate inflammatory response and insulin resistance (IR). We assessed chemerin, vaspin and leptin serum concentration and studied their association with IR laboratory and morphological features in patients with hepatitis C. The study included 40 patients with hepatitis C and 20 healthy volunteers, similar in age and body mass index (43.6 ± 11.6 vs 40.9 ± 11.8 years and 25.0 ± 4.1 vs 23.9 ± 3.3 kg/m2, respectively). Patients had to have a normal lipid profile, and diabetes was an exclusion criteria. Serum chemerin and leptin levels and IR were significantly higher in patients with hepatitis C when compared to the controls (P = 0.02, P = 0.02 and P = 0.02, respectively), whereas vaspin level was significantly decreased (P = 0.01). Serum chemerin was negatively associated with necro‐inflammatory grade (r = (–0.49), P = 0.01). The lowest levels of serum chemerin were found in patients with moderate/severe inflammation (P = 0.03). Serum leptin tended to be up‐regulated in patients with minimal inflammatory activity. Serum vaspin was higher, although not significantly, when fibrosis was more advanced. IR was positively associated with fibrosis stage (r = 0.33, P = 0.03). Serum chemerin and leptin were related to each other (r = 0.45, P = 0.02).Our findings support a complex interaction between the analysed adipokines and pathogenesis of inflammatory process in CHC. The role of chemerin and vaspin in pathogenesis of inflammatory response should be further investigated.

Breast cancer progression and expression of blood group-related tumor-associated antigens.

There is sufficient evidence that blood group related Lewis antigens are tumor-associated molecules. We have conducted immunohistochemical analysis of the expression of Lewis antigens in breast cancer tissue as an indicator of the degree of malignancy and as a prognostic factor. The studies were performed by examining 43 female patients diagnosed with invasive ductal carcinoma of the breast. Postoperative specimens were stained immunohistochemically using a panel of monoclonal antibodies (MAbs) specific for tumor-associated antigens: sialosyl LewisA, LewisA, LewisB, Lewisx, and LewisY. The aims of the study were to compare the appearance of metastases, degree of cancer stage (pTNM), and its histologic differentiation with the expression of Lewis phenotype. The evaluation of antigen expression was performed quantitatively and independently by two pathologists. Statistical significance was defined by Mann-Whitney test. The presented analysis of Lewis antigens showed higher expression of LeB and LeA (p = 0.03) in patients in stage N2 than in stage N1. The expression of LeB and LeY was higher in patients in stage T4 than in stage T1 (p = 0.02). No differences were observed for histologic differentiation. These data suggest that the expression of sialosyl-LeA and LeB antigens in breast cancer may predict metastases to lymph nodes.

Angiogenesis in chronic hepatitis C is associated with inflammatory activity grade and fibrosis stage.

Data regarding the assessment of angiogenesis in liver tissue in chronic hepatitis C (CHC) are rare. The study was performed to explain the association between the histopathological features and the number of new blood vessels in lobules and portal tracts in CHC. The second aim of the study was to define the localization of sprouting and pattern of formation of new vessels by estimating CD 34 antigen expression in the liver. The study involved 74 patients with CHC, infected with viral genotype 1b before antiviral therapy. The number of new-formatted blood vessels was positively associated with fibrosis stage and inflammatory activity grade in the liver biopsy from CHC patients. The relationship was evident in the portal tract, fibrous septa and periportal zones of lobules. The results suggest that inflammatory hepatocyte injury may promote neo-angiogenesis.

Visfatin serum levels in chronic hepatitis C patients

Summary.  Visfatin is a new adipokine involved in several processes. The data concerning visfatin in chronic hepatitis C (CHC) is small. To assess visfatin serum concentration and to study its association with biochemical and morphological features in CHC. Seventy nonobese patients with CHC (Group 1) confirmed by the presence of serum hepatitis C virus (HCV)‐RNA and 20 healthy volunteers (Group 2), similar in age and BMI with normal fasting glucose and lipid profile were included. Visfatin was significantly increased in Group 1 compared with Group 2 (55.6 ± 23.1 vs 23.7 ± 3.8 ng/mL; P < 0.001). Visfatin was negatively associated with necro‐inflammatory activity grade (r = −0.36; P = 0.007). The lowest levels were found in patients with the most advanced inflammation: grades 3–4 – 46.8 ± 17.1, grade 2 – 52.6 ± 18.4 and grade 1 – 75.2 ± 27.6 ng/mL; P = 0.017. A significant difference was also shown comparing patients with minimal inflammatory activity to the rest of the cohort (P = 0.009). Visfatin receiver operating characteristic curve analysis for different necro‐inflammatory activity – grade 1 vs grades 3–4 with area under the curve 0.81 indicated a good discriminant power for differentiation of moderate/severe inflammation, with the cut‐off set at 57.6 ng/mL (sensitivity 75%, specificity 90%, positive predictive value 0.90, negative predictive value 0.75). Serum visfatin concentration increases significantly in CHC patients. These findings suggest that visfatin is important in the pathogenesis of the inflammatory process in CHC. Visfatin may play a dual role as a pro‐inflammatory or/and protective factor. The measurement of visfatin serum concentration may serve as an additional tool in distinguishing more advanced grades of the necro‐inflammatory activity.

Hepatocyte steatosis in HCV patients promotes fibrosis by enhancing TGF‐β liver expression

Aim:  The objective of this study was to examine the relationship between TGF‐β expression in steatotic liver and the stage and yearly progression rate of fibrosis in chronic hepatitis C (CHC) patients.

Intensity of angiogenesis and mast cell infiltration in cervical intraepithelial and invasive lesions - are they correlated?

The data on the association between angiogenesis and mast cell density in cervical tumors and pretumoral conditions are scanty. The aims of the study were as follows: (1) to assess microvessel density and mast cell density in cervical lesions as well as in normal cervix samples and (2) to study the correlation between these variables. A hundred and one cervical samples were submitted to histopathological evaluation. Four study groups were distinguished: normal cervix samples, low-grade squamous intraepithelial lesions, high-grade squamous intraepithelial lesions, and invasive squamous cell carcinomas. The immunohistochemistry was performed using anti-CD34, Anti-Human Mast Cell Tryptase, and Anti-Human Mast Cell Chymase antibodies. The microvessels and mast cells in the corresponding areas of tissue samples were counted by three observers using a multi-headed microscope. Microvessel density and density of mast cells that contain tryptase increased from normal samples through intraepithelial lesions to invasive carcinoma. The density of mast cells containing chymase was significantly higher in invasive carcinomas than in normal samples. In the entire study population, but not in the separated study groups, significant correlations between microvessel density and mast cell density were found. A specific mechanism of this interaction still needs to be evaluated.

The Comparison of Scoring Scales for Liver Biopsy Assessment in Morbidly Obese Patients Undergoing Bariatric Surgery

Background: Many scoring systems have been applied for the grading and staging of non-alcoholic fatty liver disease (NAFLD). There is no consensus according to semiquantitative scales for the assessment of steatosis, inflammatory grading, and fibrosis staging in NAFLD. Methods: We analysed 24 consecutive patients who underwent bariatric surgery. The grading for steatosis was estimated according to the systems proposed by Brunt and by Dixon. Brunts scale and Scheuers scale modified by Gabriel were used for inflammatory activity and fibrosis staging. Additionally, types of NAFLD disease were diagnosed according to Matteonis classification. Results: Steatosis was observed in 88% and steatohepatitis in 54% of patients. We observed portal, periportal and pericellular fibrosis. Neither bridging fibrosis nor cirrhosis were found. Extent of steatosis estimated according to Dixon and Brunts scales was positively associated with appearance of steatohepatitis. The comparison of Dixons and Brunts scales according to grade of steatosis demonstrated a statistically significant difference. Inflammatory activity grades and fibrosis stages assessed according to Scheuer and Brunt scales differ significantly. Inflammatory activity evaluated with the Brunt scale was associated with the extent of steatosis and occurrence of steatohepatitis. Conclusions: Non-advanced forms of liver fibrosis do not appear to be dependent on steatosis and inflammatory grade in NAFLD. It is necessary to find the precise estimation of extent of steatosis especially occupying less than 1/3 or 1/4 of the lobule area. Brunts scale seems to be more useful for the estimation of liver biopsy in NAFLD. It is essential to create a consensus for evaluation of steatosis and necroinflammatory grading and fibrosis staging in NAFLD.

Prognosis of the surgical treatment of patients with non-small cell lung cancer (NSCLC) – relation to DNA ploidy

OBJECTIVE The aim of this study was to evaluate prognostic importance of cell ploidy and proliferation activity in non-small cell lung cancers. Survivals were compared according to the following factors: sex, age, histology, grading, DNA ploidy, tumour size, T factor, N factor and operative procedure. METHODS In a group of 191 patients in whom cytofluorometric examinations had been performed on archival tumour specimens, postoperative recurrences were observed. RESULTS Postoperative recurrence was observed in 64 (64.6%) of 99 patients with aneuploid tumours and in 35 (38.0%) of 92 with diploid tumours (P<0.001). Overall survival (OS) rates for the group of 92 patients operated for diploid non-small cell lung cancer (NSCLC) at 5 and 10 years were 62 and 51.1%, whereas of other 99, operated for aneuploid tumours 33.3 and 25.9%, respectively (P<0.001). In the former group of patients disease-free survival (DFS) rates at 5 and 10 years were 58.7 and 51.4% but in the latter 29.3 and 26%, respectively (P=0.00014). Significant differences dependent on cell ploidy were also observed in OS and DFS rates of patients operated respectively for SCLC (P=0.0029; P=0.00318) and adenocarcinoma (AC; P=0.0241; P=0.02109). In general, the mean percentage of S-phase cells in non-small cell lung cancers was 14.0% (SD=13.1) in patients who survived 5 years, and 22.4% (SD=15.7) in those who had a recurrence or died (P<0.001). CONCLUSIONS In our opinion the most important finding of our work is that determination of cell ploidy in NSCLC provides a valuable supplement to the TNM stage when evaluating late results of the surgical treatment. However, the paper demonstrates that aneuploidy, although unfavourable, is not an independent prognostic factor in the group of patients with NSCLC and in the subgroups - both with squamous cell carcinoma and adenocarcinoma. Our results show also that the percentage of S-phase cells is an independent, unfavourable prognostic factor in patients treated surgically for non-small cell lung cancer and in the subgroup with squamous cell lung carcinoma.

sPECAM-1 and sVCAM-1: role in pathogenesis and diagnosis of chronic hepatitis C and association with response to antiviral therapy:

Aim: To analyze the relationship between pretreatment clinical or histological features and the levels of soluble platelet-endothelial cell adhesion molecule-1 (sPECAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1), to determine their serum concentration in responders and nonresponders, to evaluate the behavior under antiviral therapy, to explain their relationship in response to therapy and to assess the association between these two molecules in chronic hepatitis C (CHC). Methods: The study analyzed 65 CHC patients, including 50 patients (Group 1) with marked fibrosis treated with peginterferon plus ribavirin, 15 patients without fibrosis (Group 2) and 13 healthy volunteers (the control group, Group 3). sPECAM-1 and sVCAM-1 levels were assessed by an immunoenzymatic method (ELISA) before and after therapy. Results: sVCAM-1 and sPECAM-1 serum concentrations increased significantly in CHC patients (p<0.001). sPECAM-1 levels corresponded to inflammatory grade (p = 0.03) and fibrosis stage (p = 0.01). sVCAM-1 increased only in advanced fibrosis. After therapy, sPECAM-1 levels decreased significantly (p<0.001) with no difference between responders and nonresponders. sPECAM-1 correlated positively with inflammatory activity (p = 0.02), fibrosis stage (p<0.001), sVCAM-1 (r = 0.56, p<0.001) and alanine aminotransferase activity (r = 0.30, p = 0.05). Receiver operating characteristic curve analysis showed a good discriminant power of serum sPECAM-1 concentrations for detection of liver fibrosis — stage 0 versus stage 1—3, AUC 0.81; cut-off 221.0 ng/ml and a fair discriminant power for distinguishing bridging fibrosis, AUC 0.78; cut-off 237.1 ng/ml. Conclusions: Hepatitis C virus (HCV) infection results in upregulation of sPECAM-1 and sVCAM-1. sPECAM-1 levels are related to necroinflammatory activity and may also identify patients with advanced fibrosis. The sPECAM-1 value was decreased by therapy but its measurement cannot predict therapy outcome and confirm HCV persistence. sPECAM-1 may influence VCAM-1 expression.

Hepatic Chemerin and Chemokine-Like Receptor 1 Expression in Patients with Chronic Hepatitis C

Introduction. Chemerin seems to be involved in pathogenesis of chronic hepatitis C (CHC). Hepatic expressions of chemerin and its receptor, chemokine receptor-like 1 (CMKLR1), in CHC have not been studied so far. Aim. To evaluate chemerin and CMKLR1 hepatic expression together with serum chemerin concentration in CHC patients and to assess their relationship with metabolic and histopathological abnormalities. Methods. The study included 63 nonobese CHC patients. Transcription of chemerin and CMKLR1 was assessed by quantitative real-time PCR, while serum chemerin was assessed by enzyme-linked immunosorbent assay. Results. Expression of chemerin and CMKLR1 was present in the liver of all CHC patients regardless of sex or age. This expression was not associated with necroinflammatory activity and steatosis grade, fibrosis stage, and metabolic abnormalities. There was a negative association between serum chemerin and chemerin hepatic expression (r = (−0.41), P = 0.006). Conclusion. The study for the first time confirmed a marked expression of chemerin and CMKLR1 in the liver of CHC patients. The study was performed using the homogenates of human liver tissue, so it is not possible to define whether hepatocytes or other cell types which are abundantly represented in the liver constitute the main source of chemerin and CMKLR1 mRNA.