Quantitative evaluation of RASSF1A methylation in the non-lesional, regenerative and neoplastic liver

Background  

Epigenetic changes during ageing and their relationship with cancer are under the focus of intense research. RASSF1A and NORE1A are novel genes acting in concert in the proapoptotic pathway of the RAS signalling. While NORE1A has not been previously investigated in the human liver, recent reports have suggested that RASSF1A is frequently epigenetically methylated not only in HCC but also in the cirrhotic liver.     

Is apoptosis a diagnostic marker of acute myocardial infarction?

OBJECTIVE: To investigate and quantify the presence of apoptosis in early myocardial ischemia in humans. METHODS: Histologic sections from the left and right ventricles of 16 hearts with impending myocardial infarction were stained with terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) method and with antibodies to p53, bcl-2, cpp32, FAS, FAS-L, and bax. DNA electrophoretic analysis was also performed. RESULTS: According to the inclusion criteria, all 16 cases showed morphologic changes consistent with ischemia and/or reperfusion. TUNEL results were positive in 14 of the 16 ischemic areas. Unexpectedly, they were also positive in "remote from ischemia" myocardium of both the left and right ventricles. DNA electrophoretic analysis confirmed the results of TUNEL. Immunohistochemistry was uniformly negative, probably because of autolysis phenomena. CONCLUSIONS: We showed that apoptosis precedes necrosis in humans, but the detection of apoptosis cannot be used as a diagnostic tool, since it can also be triggered by nonischemic events.     

A new mathematical formula for predicting long bone length in early pregnancy.

OBJECTIVE: To propose new mathematical formulae to estimate fetal long bone biometry in early pregnancy and to establish their efficacy in comparison to previously constructed mathematical formulae. METHODS: A study population of 1960 singleton euploid fetuses was referred for transvaginal ultrasound examinations between 71 and 112 days of gestation prior to genetic amniocentesis. To determine the relationship between the biparietal diameter and long bone length, a sample group of 400 randomly chosen normal fetuses was evaluated. Regression equations were derived, then tested in the remaining 1560 control fetuses and compared with previously reported mathematical formulae by other authors. Mean absolute error, mean absolute percentage error and mean systematic error with their standard deviations were calculated. RESULTS: The relationships between femur or humerus length vs. biparietal diameter (BPD) and gestational age (GA) were, respectively: expected femur length = -16.92108 + 0.4569402 x BPD + 0.171617 x GA (P < 0.001) and expected humerus length = -16.28531 + 0.4283019 x BPD + 0.1696017 x GA (P < 0.001). The confidence intervals of the predicted values for different values of biparietal diameter and gestational age and confidence intervals for the regression coefficients, such as the distribution of the residuals, are given. All previous formulae obtained by transabdominal ultrasound demonstrated an overestimation of expected long bones measurements; this was reduced using different formulae obtained in early pregnancy. Using our mathematical formulae, the mean absolute percentage error and the mean systematic error in estimating femur and humerus length were very low (11.15% and -2.02%; 10.59% and -1.74%, respectively). CONCLUSIONS: The new ultrasonographic morphometric models derived from transvaginal measurements in early pregnancy show a good reliability in estimating fetal long bone length.     

Immunomodulatory effects of the fungicide Mancozeb in agricultural workers

Available data suggest that ethylenebisdithiocarbamates (EBDCs) may have immunomodulatory effects. This study aimed to investigate the immunological profile of farmers exposed to Mancozeb, an EBDC fungicide, through the determination of several serum, cellular, and functional immune parameters. Twenty-six healthy subjects entered the study, 13 vineyards exposed to Mancozeb and 13 unexposed controls. Exposure was assessed through the determination of ethylentiourea (ETU) in urine. Complete and differential blood count, serum immunoglobulins, complement fractions, autoantibodies, lymphocyte subpopulations, proliferative response to mitogens, natural killer (NK) activity, and cytokine production were measured. Post-exposure samples showed ETU urine concentration significantly higher than pre-exposure and control groups. A significant increase in CD19+ cells, both percentage and absolute number, and a significant decrease in the percentage of CD25+ cells were found in post-exposure samples compared to controls. A statistically significant increase in the proliferative response to phorbol myristate acetate plus ionomycin (PMA + ionomycin) was observed in the post-exposure group compared to controls and baseline, while a significant reduction in LPS-induced TNF-alpha release in post-exposure samples was observed. Overall, our results suggest that low-level exposure to Mancozeb has slight immunomodulatory effects, and point out a method adequate to reveal immune-modifications in workers occupationally exposed to potential immunotoxic compounds, based on a whole blood assay.     

Bile duct injury during laparoscopic cholecystectomy: results of an Italian national survey on 56 591 cholecystectomies

HYPOTHESIS: Bile duct injury (BDI) remains the most serious complication of cholecystectomy. With laparoscopic cholecystectomy (LC), the incidence has become more frequent. This study verifies the current incidence, mechanism, presentation, and treatment of BDI occurring during LC in general surgical practice. DESIGN: Anonymous retrospective multicenter survey. SETTING: Department of surgery at a university referral center, collecting data from general surgical units. PATIENTS: Data from 56 591 patients who underwent LC between January 1, 1998, and December 31, 2000, in 184 hospitals in Italy were analyzed. MAIN OUTCOME MEASURES: Current incidence, mechanism, presentation, and treatment of BDI occurring during LC in general surgical practice. RESULTS: Two hundred thirty-five BDIs were reported, with an overall incidence of 0.42%. There were no risk factors in 80.0% of the patients. Poor identification of the anatomical features of the hepatic pedicle was the most frequently reported cause (36.8%), and technical problems accounted for 27.0% of causes. The incidence of BDI was higher during cholecystitis (P<.001) and decreased with increasing number of LCs performed by the surgical teams (P<.01). There was no difference in incidence according to technique (French or US) or to routine or selective intraoperative cholangiography. One hundred eight BDIs (46.0%) were recognized intraoperatively and immediately repaired in 89.8% of patients. One hundred twenty-seven BDIs (54.0%) were diagnosed postoperatively, the dominant manifestation being biliary fistula (44.1%). CONCLUSIONS: This study confirms a higher incidence of BDI during LC. It highlights the relevance of the number of previously performed LCs and of the correct surgical technique to avoid BDI. The need for correct procedures, adequate expertise of the repairing surgeon in BDI repairs, and a multidisciplinary approach in the management of BDI is emphasized.     

Effect of inhibitors of choline acetylation on acetylcholine output and motility in response to anticholinesterases and to distension of the lumen of isolated guinea-pig ileum

Some pharmacological effects of acyl-aryl compounds, which inhibit the enzymatic acetylation of choline, have been studied on the isolated guinea-pig ileum. Diphenylylbutylacetic acid (M.G. 1763) greatly reduced the output of acetylcholine from intestine. The intestinal contractions produced by anticholinesterase drugs were inhibited, while the response to acetylcholine itself was little affected. The contraction of longitudinal muscle elicited by raising intra-luminal pressure was also decreased, while the emptying phase of the peristaltic reflex was scarcely affected. There was a correlation between inhibitory activity and chemical structure.