Screening for nonalcoholic fatty liver disease in patients with type 2 diabetes mellitus using transient elastography - a prospective, cross sectional study.

AIM: To investigate the prevalence and severity of nonalcoholic fatty liver disease (NAFLD) in patients with diabetes mellitus type 2 (T2DM), based on increased controlled attenuation parameter (CAP) and liver stiffness measurements obtained by transient elastography. In addition, we aimed to identify parameters that correlate with increased elastographic parameters of steatosis and fibrosis to provide a better indication when a patient with T2DM should be screened for NAFLD. METHODS: We conducted prospective, cross-sectional study of 679 consecutive adult patients with diagnosed T2DM mean age 65.2±11.6. NAFLD was defined by transient elastography. In 105 patients a percutaneous liver biopsy (LB) was done. RESULTS: The prevalence of NAFLD based on transient elastography was 83.6%. Independent factors associated with increased CAP were higher body mass index, longer T2DM duration, higher serum triglyceride, lower levels of vitamin D, higher C-reactive protein, and higher HOMA-IR. The prevalence of moderate liver fibrosis was 26.9% and advanced liver fibrosis 12.6%. Independent factors associated with moderated fibrosis based on elastography were higher body mass index and higher levels of alanine aminotransferase (ALT), while independent factors associated with advanced fibrosis were female gender, higher body mass index, higher levels of ALT, gama-glutamil transferase and C-reactive protein. Sixty-four (60.9%) of 105 patients with LB had NAFLD activity score ≥5. Regarding the presence and stages of fibrosis based on LB, moderate fibrosis was found in 29.5% of patients, while 29.5% had advanced fibrosis and 6.7% cirrhosis. CONCLUSION: This study supports more aggressive screening for NAFLD and fibrosis in patients with T2DM.

Prospective evaluation of non-alcoholic fatty liver disease by elastographic methods of liver steatosis and fibrosis; controlled attenuation parameter and liver stiffness measurements.

AIMS: To examine the temporal changes of both controlled attenuation parameter (CAP) and liver stiffness measurements (LSM), assessed by Fibroscan, in a large sample of patients with non-alcoholic fatty liver disease (NAFLD). METHODS: In this prospective, observational study, we consecutively enrolled 507 adult individuals with Fibroscan-defined NAFLD who were followed for a mean period of 21.2 ±â€¯11.7 months. RESULTS: During the follow-up period, 84 patients (16.5%) had a progression of CAP of at least 20% with a median time of 39.93 months, while 201 (39.6%) patients had a progression of LSM of at least 20% with median time of 30.46 months. There were significant differences in the proportion of LSM progression across body mass index (BMI) categories, with obese patients having the highest risk of progression over the follow-up (hazard ratio 1.66; 95%CI 1.23-2.25). Multivariable regression analysis showed that BMI and serum creatinine levels were the strongest predictors for CAP progression in the whole population, while HOMA-estimated insulin resistance was an independent predictor of LSM progression over time in the subgroup of obese patients. CONCLUSION: This prospective study shows for the first time that the progression risk of both liver steatosis and fibrosis, detected non-invasively by Fibroscan, is relevant and shares essentially the same metabolic risk factors that are associated with NAFLD progression detected by other invasive methods.

Comparison of Surgisis, Vypro II and TiMesh in contaminated and clean field.

PURPOSE: The study aimed to evaluate the histologic properties and infection resistance of three different mesh materials in a rat model. METHODS: Each mesh, in both infectious (n = 96) and non-infectious groups (n = 270), was positioned both in sublay (preperitoneally) and onlay (subcutaneously) locations. Properties of the biological (Surgisis; Cook Surgical), composite, partially resorbing (Vypro II mesh; Ethicon) and non-resorbing (TiMesh; GFE Medizintechnik GmbH) mesh were evaluated and compared. Animals were killed at 7, 21 and 90 days after implantation. The following parameters were evaluated to assess the host response to the mesh material: inflammation, vascularization, fibrosis, collagen formation, Ki67, and a foreign body reaction by granuloma formation (FBG). RESULTS: Surgisis mesh produced more pronounced inflammation and cell proliferation, and less intense granuloma formation, as well as fibrosis, compared to the other two groups. When the infected materials were examined, we found signs of local infection to be more often present in Surgisis group of animals. CONCLUSIONS: In the presence of bacterial contamination, no benefits were observed in the use of the Surgisis prosthesis over the use of TiMesh and Vypro II.

MAGE-A3/4 and NY-ESO-1 antigens expression in metastatic esophageal squamous cell carcinoma.

In the present study we analyzed immunohistochemical expression of MAGE-A 3/4 and NY-ESO-1 in 55 samples of esophageal squamous cell carcinomas (ESCC) and their respective lymph node metastases. To our knowledge this is the first study to assess and compare the expression of these antigens in ESCC lymph node metastases. Fifty (90.9%) primary ESCC were positive for MAGE-A 3/4 and 53 (96.6%) were positive for NY-ESO-1. MAGE-A 3/4 was expressed in all lymph node metastases and the intensity of expression was high in a majority of cases. NY-ESO-1 was negative in 2 (7.1%) lymph nodes metastases, while the reaction was predominantly moderate in the positive group. In primary tumors MAGE-A 3/4 showed a significantly higher intensity of expression compared to NY-ESO-1 (P=0.047), while in lymph node metastases the intensity of expression was not significantly different (P=0.387). Primary tumors with and without lymph node metastases showed no significant differences in MAGE-A 3/4 (P=0.672) and NY-ESO-1 (P=0.444) expression. Intensity of MAGE-A 3/4 (P=0.461) and NY-ESO-1 (P=0.414) expression in primary tumors was not significantly different compared to the expression in their respective lymph nodes metastases. Expression of MAGE-A 3/4 in primary tumors showed significant positive correlation with primary tumor expression of NY-ESO-1 (P=0.021) but no significant correlation with the expression of MAGE-A 3/4 in lymph node metastases (P=0.056). Expression of NY-ESO-1 in primary tumors showed significant positive correlation with the expression of NY-ESO-1 in lymph node metastases (P=0.001) and significant negative correlation with patients’ age (P<0.001). Expression of MAGE-A 3/4 and NY-ESO-1 in primary tumors and lymph node metastases showed no significant correlation with prognostic parameters such as tumor grade and TNM stage (P>0.05). We have shown different levels of MAGE-A 3/4 and NY-ESO-1 expression in almost all specimens of primary tumor and lymph node metastases, suggesting that ESCC may be possible target of immunotherapy and anti-tumor vaccination. High levels of expression in lymph node metastases indicate possible clinical benefit of postoperative vaccine with MAGE-A3 and NY-ESO-1 in advanced stage of disease.

Laparoscopic cholecystectomy in an ambulatory setting--initial experience in Croatia.

BACKGROUND/AIMS: The purpose of this study was to discover if ambulatory laparoscopic cholecystectomy is feasible in our environment and to determine the factors which will predict its success. METHODOLOGY: From November 12th 2002, to February 25th 2005 one hundred and fifty patients were admitted to our surgical ward of the General Hospital in Bjelovar due to cholelithiasis or biliary colic. Patients were unselected. Six to eight hours after the surgery we rated the ability of the patients to be discharged from the hospital (although they weren't discharged). RESULTS: The research has shown that 61% of the patients were eligible for ambulatory procedure. Among ultrasonographic attributes the most significant independent predictive factor of potential complications, prolongation of the procedure and ineligibility of the patient for the ambulatory laparoscopic cholecystectomy in our study was the thickened wall of the gallbladder. Among non-ultrasound preoperative attributes, the most important are the anamnestic data of prior cholecystitis and ASA classification. Pain was the most common reason why the patients were classified as non-candidates (36.6%). In the second place (25.4%) was the negative subjective patient's judgement to be discharged when all objective parameters were good. CONCLUSIONS: LC is also feasible in our environment but the patient's own motivation is the key factor in our situation.

Laparoscopic cholecystectomy and perioperative ERCP.

BACKGROUND/AIMS: Study of acceptance of simultaneous laparoscopic cholecystectomy and endoscopic retrograde cholangiopancreatography for treatment of cholelithiasis with choledocholithiasis. METHODOLOGY: There were 25 patients. Ten patients had acute pancreatitis of biliary etiology, accompanied by transitory icterus. In 15 patients, choledocholithiasis was suspected preoperatively both on ultrasonography and i.v. cholangiography. In all patients laparoscopic cholecystectomy with perioperative endoscopic retrograde cholangiopancreatography and sphincterotomy were performed for the treatment of cholelithiasis and choledocholithiasis. RESULTS: Simultaneous laparoscopic cholecystectomy and endoscopic retrograde cholangiopancreatography was successfully done in all patients. The patients were discharged home on the 4th day after the surgery. Concerning early complications, there where 3 early complications, e.g., prolonged hemorrhage after papillotomy in a patient with choledocholithiasis with stenotic papillitis. Conservative therapy (fresh frozen plasma, local hemostats) was used in this patient. In 4 patients with choledocholithiasis, transitory hyperamylasemia was observed, with no clinical symptoms of pancreatitis. The symptoms disappeared with conservative therapy 3 days after the beginning of treatment. CONCLUSIONS: Simultaneous laparoscopic cholecystectomy and endoscopic retrograde cholangiopancreatography for treatment of cholelithiasis and choledocholithiasis is a safe and acceptable treatment.

Estimation of genome damage after exposure to ionizing radiation and ultrasound used in industry.

The application of ionizing radiation in industry for nondestructive testing entails a specific framework of working conditions that include field work, facilities with different radioactive sources, maintenance thereof without halting production, use of nonionizing radiation, and exposure to chemical agents. The present study gives an estimation of recent genome damage in two groups of subjects using chromosome aberration assay and micronucleus assay. The first group was exposed to (192)Ir and the second was simultaneously exposed to (192)Ir and ultrasound. The results show that both groups had higher values of chromosome aberrations and micronucleus frequency than controls. The group of examinees exposed both to (192)Ir and ultrasound had significantly more chromatid breaks, acentric fragments, and dicentric chromosomes, and had a significantly higher frequency of micronuclei than subjects exposed to (192)Ir only. The study suggests that the detected differences in the genome damage may be attributed to the action of ultrasound. This study confirms the dosimetry data for ionizing radiation, which indicate that the methods used in industrial radiography and the usage of nonionizing radiation entail an increased health risk. In the absence of personal dosimeters for nonionizing radiation and chemical agents, biomonitoring provides reliable parameters for estimation of genome damage and may lead to improvements in working conditions and radiation safety programs.

Safety of simultaneous resections of colorectal cancer and liver metastases.

Liver resection is the only potentially curative method for patients with colorectal cancer metastases and 5-year survival rates are 20%-40%. Simultaneous resection of colorectal cancer and synchronous liver metastases has been recommended if minor hepatectomy is indicated. The purpose of this paper is to analyze the treatment of hepatic colorectal secondaries and to assess the safety of simultaneous and delayed liver resections and relations of morbidity to the extensiveness of hepatectomy and perioperative factors. Analyzed were 21 patients with liver metastases from colorectal cancer operated between 1997 and 1999 in the Clinical Hospital "Sestre milosrdnice". Operating time for simultaneous colorectal and liver resections was not significantly longer compared to liver resections alone. No significant difference in complication rate was found after simultaneous procedures and liver resection alone (38% vs. 31%). Complication rate after major liver resections was not significantly greater than after minor resections (38% vs. 31%). No statistically significant differences were found in operation time and blood replacement between patients who developed postoperative complications and those who did not. In conclusion, simultaneous resections of primary colorectal cancer and liver metastases may be considered safe. Morbidity rates are not significantly different from those after liver resections alone, nor depend significantly upon the extensiveness of liver resection, providing that the operation time and blood loss are within the range observed in this study.

[In vitro and in vivo micronucleus tests in genotoxicity research]. / In vitro i in vivo mikronukleus metode u genotoksikoloskim istrazivanjima.

Working and living environment today is an unpredictable mixture of different chemical substances. Some of these chemicals are mutagenic carcinogens and others are aneugenic carcinogens. An additional burden to the human genome is the exposure to several kinds of radiations. Among a dozen genotoxicological methods in use in vitro and in vivo, the micronucleus assay has found application in the basic research, clinical research, and pharmacological studies. The advantage of in vivo and in vitro micronuclues assays is in the fact that they provide data on both clastogenic and aneugenic action of agents. Although several years behind the in vitro technique, the in vivo micronucleus assay has swiftly developed into a significant source of data in acute and chronic genotoxic investigations, as it does not require cell culture.

Cytogenetic consequences after occupational exposure to antineoplastic drugs.

Cytogenetic monitoring was carried out on a group of 38 nurses who reconstitute antineoplastic drugs in order to determine the extent of chromosomal damage. Genotoxic activities of antineoplastic drugs are studied by chromosome aberration assay, micronucleus assay, sister chromatid exchange (SCE) frequency high frequency cells (HFC) analysis, and mitotic activity of peripheral lymphocytes. Results confirmed that occupational exposure to a mixture of antineoplastic drugs may cause genome damages. The results of this study show that biomonitoring after exposure to a mixture of antineoplastic drugs which express clastogenic and aneugenic activity should involve a battery of cytogenetic methods.

Apoptosis in human gastric polyps and adenocarcinomas: a stereological analysis.

BACKGROUND/AIM: The occurrence of apoptotic cells was analyzed in human normal gastric mucosa, polyps and adenocarcinomas. METHODOLOGY: Histological classification was carried out on hematoxylin and eosin stained slides. The tissue was classified as follows: Normal gastric mucosa or adenomatous polyps. Gastric carcinoma specimens were histologically classified according to Lauren's classification into: A: Diffuse adenocarcinoma without metastasis, B: Diffuse adenocarcinoma with metastasis, C: Intestinal adenocarcinoma without metastasis, D: Intestinal adenocarcinoma with metastasis, E: Mixed adenocarcinoma without metastasis and mixed adenocarcinoma with metastasis. The counting of apoptotic cells was performed using the 40X objective with a calibrated eyepiece Weibel's multi-purpose M 42 stereological test system. Each group was evaluated stereologically, determining numeric density of apoptotic cells. RESULTS: The results show the progressive and statistically significant increase of apoptotic numeric densities from normal gastric epithelium to adenomatous polyp and finally to cancer, which contained the highest number of apoptotic cells. Comparing gastric carcinoma with and without metastasis in intestinal and diffuse adenocarcinoma there was statistically significant difference. In these two groups, carcinomas with metastasis contained higher number of apoptotic cells than without metastasis. Gastric cancer according to numeric densities of apoptotic cells can be separated in tree statistically different groups: A: Intestinal type gastric cancer with metastasis (the highest number of apoptotic cells), B: Intestinal type gastric cancer without metastasis and diffuse gastric cancer with metastasis (medium number), C: Diffuse type gastric cancer without metastasis, mixed gastric cancer with and without metastasis (the lowest number of apoptotic cells). CONCLUSION: These results suggest that numeric densities of apoptotic cells are associated with tumor progression in human gastric carcinogenesis and can be used as prognostic mark.

A nine-year follow up study of a population occupationally exposed to vinyl chloride monomer.

The presence of structural chromosome aberrations and sister chromatid exchange frequencies (SCE) were studied in lymphocytes from 28 male subjects occupationally exposed to vinyl chloride monomer for a period of 9 years. A significant increase in chromosomal damages and elevated SCE frequencies was detected during the third and fourth year of the follow-up study. During the last 2 years, all examined parameters are approaching control values. This is a result of decreasing vinyl chloride monomer concentrations in the working environment without interrupting the working process.

Helicobacter pylori in a group of endoscopically examined patients in the county of Medimurje.

The presence of Helicobacter pylori was investigated in 50 patients, mean age 54 years and age range 28-56 years. Gastroduodenoscopy and biopsy of the antral and/or pyloric part of gastric mucous membrane were performed in all study patients. Bioptic tissue was examined by culture and histologic staining, and tested by a rapid urease test. According to overall results, the urease test was most sensitive, i.e. positive in 23 (45%) patients, whereas histological staining was positive in 14 (29) patients. Endoscopic diagnosis revealed the following: duodenal ulcer--histologic stain 3/5 (55%), urease test 9/21 (43%); gastric erosion--histologic stain 4/13 (31%), urease test 7/13 (55%); gastroduodenitis--histologic stain and urease test 2/4 (50%). According to endoscopy, positive pyloriset test (Orion Diagnostica) was found as follows: duodenal ulcer 5/7 (71%), gastric ulcer 8/10 (80%), gastric erosion 10/12 (83%) and gastroduodenitis 5/7 (71%). The authors recommend the diagnosis of Helicobacter pylori as a routine approach in gastroenterological routine.

[Scanning electron microscopy analysis of peroperative injury at the junctions of the vasa vasorum and the collateral veins of venous autografts prepared for myocardial revascularization]. / Scann-elektronskomikroskopska analiza peroperativnih ostecenja utoka vasa vasorum i kolateralnih vena venoznog autotransplantata pripremljenog za revaskularizaciju miokarda.

The authors' objective was to determine the peroperative changes in affluences of collateral veins and vasa vasorum of venous autografts prepared for a coronary bypass by standard and proposed methods. The investigations were carried out on dogs. In control group, the veins were prepared by the standard method where the vein is presented by several minor incisions with "skin bridges" planted in between. In the second group, the veins were prepared by the method developed by the authors, where the vein is presented by a long incision, maintaining perfusion with its own blood throughout surgery. The scanning electron microscope analysis has shown that the standard method of vein preparation causes significant damage in collateral veins and vasa vasorum affluences which are actually part of an internal surface of future venous autograft prepared for myocardium revascularization. The proposed surgical method considerably reduces possible peroperative damage of the vasa vasorum and collateral vein affluence sites forming a portion of the internal surface of the venous autograft. Therefore, based on the obtained results, this method is recommended for clinical application.