Circulating levels of cytochrome C, gamma-glutamyl transferase, triglycerides and unconjugated bilirubin in overweight/obese patients with non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease, characterized by hepatocyte apoptosis, is distinct in fatty liver and non-alcoholic steatohepatitis, the more severe form. Apoptotic cell death is caspase-dependent and associated with mitochondrial membrane depolarization and cytochrome c release. Adhering to the hypothesis that the exposure of hepatocytes to free fatty acids, resulting in increased ROS production and mitochondrial damage, is balanced by the presence of antioxidant substances, circulating levels of gamma-glutamyl transferase, cytochrome c, triglycerides and unconjugated bilirubin were explored in patients suffering from non-alcoholic fatty liver disease with different severity. One hundred and eighty-six consecutive patients who presented recent ultrasound feature of bright liver without any liver disease of known origin were enrolled, eighty-nine of whom underwent liver biopsy. Forty-five subjects were allocated on the basis of histology in fatty liver group while 44 patients formed the group with non-alcoholic steatohepatitis. A cohort of 27 young, lean, apparently healthy individuals was selected as control group. The levels of gamma-glutamyl transferase were normal or slightly increased, while unconjugated bilirubin concentrations were elevated in all the spectra of non-alcoholic fatty liver disease. Comparing the present results with relevant findings from other studies dealing with diseases characterized by apoptosis, we did not find high circulating levels of cytochrome c in non-alcoholic fatty liver disease. What is more, our patients, categorized as suffering from simple fatty liver or from the more severe non-alcoholic steatohepatitis, had similar levels of cytochrome c and gamma-glutamyl transferase, p=0.19 and 0.11. Serum triglycerides were higher in patients with non-alcoholic fatty liver disease than in the healthy group, p=0.001. These findings likely reflect a balance between oxidative stress and anti-oxidant response rather than a lack of reliability of cytochrome c as a reliable biomarker of mitochondrial damage.

The carbon footprint of dairy production systems through partial life cycle assessment.

Greenhouse gas (GHG) emissions and their potential effect on the environment has become an important national and international issue. Dairy production, along with all other types of animal agriculture, is a recognized source of GHG emissions, but little information exists on the net emissions from dairy farms. Component models for predicting all important sources and sinks of CH(4), N(2)O, and CO(2) from primary and secondary sources in dairy production were integrated in a software tool called the Dairy Greenhouse Gas model, or DairyGHG. This tool calculates the carbon footprint of a dairy production system as the net exchange of all GHG in CO(2) equivalent units per unit of energy-corrected milk produced. Primary emission sources include enteric fermentation, manure, cropland used in feed production, and the combustion of fuel in machinery used to produce feed and handle manure. Secondary emissions are those occurring during the production of resources used on the farm, which can include fuel, electricity, machinery, fertilizer, pesticides, plastic, and purchased replacement animals. A long-term C balance is assumed for the production system, which does not account for potential depletion or sequestration of soil carbon. An evaluation of dairy farms of various sizes and production strategies gave carbon footprints of 0.37 to 0.69kg of CO(2) equivalent units/kg of energy-corrected milk, depending upon milk production level and the feeding and manure handling strategies used. In a comparison with previous studies, DairyGHG predicted C footprints similar to those reported when similar assumptions were made for feeding strategy, milk production, allocation method between milk and animal coproducts, and sources of CO(2) and secondary emissions. DairyGHG provides a relatively simple tool for evaluating management effects on net GHG emissions and the overall carbon footprint of dairy production systems.

Does spleen volume play a role in patients with HCV-related chronic hepatitis?

As the lymphotropism of hepatitis C virus (HCV) has already been ascertained, and in the light of the fact that the immune defense system is an organized network composed of functionally interrelated tissues, this study was carried out to verify the possible involvement of spleen in HCV-related chronic hepatitis. In this cross-sectional study we measured spleen longitudinal diameter by ultrasound, beta2-microglobulin serum levels and splenic artery resistivity index (SARI) by Doppler in 51 patients treated with standard combined (Peg-Interferon plus Ribavirin) antiviral therapy. Thirty-three patients (17 females) completed the regimen and were compared to 31 controls (16 females). The mean basal values of spleen longitudinal diameter were higher in patients with chronic hepatitis than in controls, i.e., 116 mm (9.4) versus 102.7 mm (9.3), P = 0.0001. In the same patients a significant trend towards increased spleen longitudinal diameter was found after antiviral therapy, independently of the stage of HCV-related chronic hepatitis. The median values of the beta2-microglobulin concentrations were not significantly higher in the patients with HCV-related chronic hepatitis compared to controls, i.e., 1.3 (0.5-2.6) versus 1 (0.6-1.4), P = 0.16, although during the course of therapy they were significantly increased. SARI values of HCV-related chronic hepatitis patients were different from those of controls, but were unvaried compared to values at the end of treatment. Neither spleen measurements nor serum beta2-microglobulin levels were able to predict therapeutic response to antiviral therapy. A stimulation/expansion of lymphoid tissue was found in patients with HCV-related chronic hepatitis. Such evidence raises the question whether physicians should continue to prescribe antiviral therapy in non-responders and supports the use of a new scheme (SLD plus beta2-MG) to diagnose this ongoing, apparently reversible, but nevertheless dangerous immunologic process.

Analysis of c-erbB-2 protein expression in conjunction with DNA content using multiparameter flow cytometry.

Breast cancer is a leading cause of cancer death among women. Factors useful for determining the prognosis of breast cancer include axillary lymph node involvement, tumor size, hormonal receptor status, nuclear grade, and relative DNA content. The c-erbB-2 protooncogene is amplified in 10-40% of primary breast tumors, as well as in breast cancer cell lines; where it is amplified there is increased expression of its product. We have investigated the DNA content and c-erbB-1 protein expression in tumor cell lines and in breast cancer patient specimens by multiparameter flow cytometry. The study was enabled by the discovery that both cellular integrity and c-erbB-2 antigen reactivity were preserved in cells and tissues following fixation in 70% ethanol. We demonstrate that flow cytometric analysis of c-erbB-2 expression in populations of ethanol-fixed tumor cells is a reliable and sensitive quantitative method that correlates well with previously documented semiquantitative techniques. This is a feasible method for analyzing archived clinical samples, and further allows correlations between c-erbB-2 levels and other cellular parameters. Additionally, this method detects abnormal populations not identified by DNA content analysis alone. Further studies utilizing this approach are necessary to evaluate the prognostic value of this oncoprotein in human breast cancer.

Cell- and tissue-specific expression of a 34,000-molecular-weight peptide growth factor in humans.

A 34,000-dalton peptide growth factor that we originally identified in human placental trophoblasts and in certain carcinomas was shown to be expressed in several normal human tissues. A highly specific antibody to the trophoblast-derived growth factor was used in an immunoperoxidase staining technique to identify the immunoreactive peptide in tissue sections. Immunoreactivity was seen in the adrenal cortex, Leydig cells of the testes, and follicular cells of the thyroid. In addition, strong staining was seen in the ducts and terminal ductules of the pancreas, in glandular epithelium of the prostate, in the chief cells of the stomach, and in the columnar epithelium of the trachea and bronchus of the lung. Certain tissues were negative for the peptide, including the adrenal medulla, liver, esophagus, small intestine, colon, bladder, lymph node, spleen, bone marrow, and thymus. Thus, the expression of the peptide depends on cell lineage and the state of differentiation; tissues of hematopoietic lineage are devoid of the 34,000-dalton peptide, whereas some of the major hormone-secreting tissues that are under pituitary control show the highest immunoreactivity.

A specific antibody to a new peptide growth factor from human placenta: immunocytochemical studies on its location and biosynthesis.

Recently, we isolated a new peptide growth factor of Mr 34 000 from synctial membranes of human placenta. In its polypeptide molecular weight and receptor binding specificity it is unlike several known growth factors. In this paper we described immunocytochemical studies on its cellular location and biosynthesis. A rabbit antiserum was raised against a homogeneous preparation of the placental peptide. The specificity of the antibody was established by immunoprecipitation and immunoblot analyses. The antibody recognized both the native and denatured 34-kilodalton (kDa) peptide but showed no binding to a variety of other growth factors and hormones tested. The antibody was used to investigate the genesis and location of the 34-kDa membranous mitogen. Immunoperoxidase staining of placental tissue slices revealed a restricted localization of the antigen in the cytoplasmic organelles of cytotrophoblasts and in the brush border membranes of syncytiotrophoblasts. No other placental structures contained the antigen. A developmentally regulated appearance of the mitogen was suggested by the fact that first trimester placenta consistently stained far more strongly than term placenta. These studies show that the 34-kDa mitogenic protein originates in placenta from embryo-derived cellular structures and suggest that in its strategic location it may influence trophoblastic growth in an autocrine manner. In other studies we investigated the presence and biosynthesis of the 34-kDa peptide in the A431 vulval carcinoma cell line, which was shown earlier to contain a membrane-associated 34-kDa growth factor. The studies demonstrate that this cell line, as well as some other human carcinomas of breast and bladder origin, actively expresses this peptide.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential diagnosis between mesotheliomas and metastatic adenocarcinomas using monoclonal antibodies against gastrointestinal carcinoma antigen and stage-specific embryonic antigen.

Monoclonal antibodies made against gastrointestinal carcinoma antigen (GICA) and stage specific embryonic antigen (SSEA) were evaluated for their ability to distinguish normal mesothelial cells present in pleural and peritoneal fluids from adenocarcinoma cells in tissue and cytology specimens. The presence of GICA was documented in a high percentage of adenocarcinomas from the gastrointestinal tract (75/98) and in 52% of pulmonary (15/29) and 29% of ovarian (6/21) adenocarcinomas. GICA was found infrequently in breast carcinoma (1/18) and not in mesotheliomas (0/16). A similar pattern of GICA expression was seen in malignant effusions from adenocarcinomas (18/47) and mesotheliomas (0/6). SSEA was found in a high percentage of adenocarcinomas derived from the gastrointestinal tract (47/56) and the lung (26/29). SSEA was detected in breast carcinoma (8/15) more often than GICA. SSEA was detected rarely in mesotheliomas (1/16). Reactivities for epithelial membrane antigen, keratin, carcinoembryonic antigen, GICA and SSEA in adenocarcinoma and mesotheliomas were compared.