We report on the results of high-resolution spec-troscopic studies on the 4-hydroxystyrene-CO(2) cluster. We show that these clusters are generated upon heating of para-coumaric acid, the chromophore of the photoactive yellow protein (PYP), as the result of a thermal decarboxylation process. Since the mass of the cluster and the starting material are the same, standard mass-resolved multiphoton ionization spectroscopic methods do not suffice to distinguish these clusters from para-coumaric acid. Instead, more advanced methods that include various UV and IR depletion methods need to be applied. These methods, in combination with quantum chemical calculations, enable us to unravel the structural and spectroscopic properties of 4-hydroxystyrene-CO(2) as well as of its hydrate, 4-hydroxystyrene-CO(2)-H(2)O.

A dual isotype (IgG, IgA) enzyme-linked immunosorbent assay (ELISA) designed to provide enhanced detection of primary biliary cirrhosis (PBC)-specific autoantibodies against both major mitochondrial and nuclear antigens has been developed and recently become commercially available. The assay (PBC Screen) simultaneously detects IgG and IgA autoantibodies to the immunodominant portions of the 3 major mitochondrial (MIT3) and nuclear (gp210, and sp100) antigens. The aim of this study was to compare the performance of the PBC Screen to the combined performance obtained with individual IgG ELISAs to MIT3, gp210, and sp100 on a large group of selected patients from multiple centers. A total of 1175 patients with PBC and 1232 subjects without PBC were evaluated. Non-PBC groups included healthy controls (624) as well as individuals with autoimmune hepatitis (281), primary sclerosing cholangitis (77), viral hepatitis (91 hepatitis B and 98 hepatitis C), other liver diseases (31), and other infectious or autoimmune diseases (30). The PBC Screen at the receiver operator characteristic optimized cutoff of 27.8 units, had an overall sensitivity of 83.8%, specificity of 94.7% and area under curve of 0.9212. This was similar to the specificity of 96.1% obtained by the combined results of individual MIT3, sp100, and gp210 IgG ELISAs (kappa index at 0.898). Of the 253 PBC patients without AMA detectable by immunofluorescence, 113 (44.7%) were interpreted as positive for PBC-specific autoantibodies. In conclusion, the PBC Screen is an appropriate first-line test for the diagnosis of PBC, including for patients negative for markers assessed using conventional methods.

Background. Risk-adjusted mortality rates are used to compare quality of care of different hospitals. We evaluated the EuroSCORE (European System for Cardiac Operative Risk Evaluation) in patients undergoing isolated coronary artery bypass grafting (CABG).Patients and method. Data of all CABG patients from January 2004 until December 2008 were analysed. Receiver-operating characteristics (ROC) curves for the additive and logistic EuroSCOREs and the areas under the ROC curve were calculated. Predicted probability of hospital mortality was calculated using logistic regression analyses and compared with the EuroSCORE. Cumulative sum (CUSUM) analyses were performed for the EuroSCORE and the actual hospital mortality.Results. 5249 patients underwent CABG of which 89 (1.7%) died. The mean additive EuroSCORE was 3.5+/-2.5 (0-17)(median 3.0) and the mean logistic EuroSCORE was 4.0+/-5.5 (0-73)(median 2.4). The area under the ROC curve was 0.80+/-0.02 (95% confidence interval (CI) 0.76 to 0.84) for the additive and 0.81+/-0.02 (0.77 to 0.85) for the logistic EuroSCORE. The predicted probability (hazard ratio) was different from the additive and logistic EuroSCOREs. The hospital mortality was half of the EuroSCOREs, resulting in positive variable life-adjusted display curves. Conclusions. Both the additive and logistic EuroSCOREs are overestimating the in-hospital mortality risk in low-risk CABG patients. The logistic EuroSCORE is more accurate in high-risk patients compared with the additive EuroSCORE. Until a more accurate risk scoring system is available, we suggest being careful when comparing the quality of care of different centres based on risk-adjusted mortality rates.(Neth Heart J 2010;18:355-9.).

HASH(0x1ce60850)

Liver cancer, especially hepatocellular carcinoma (HCC), is one of the most common tumors worldwide. The majority of people with HCC will die within 1 year of its detection. The high case-fatality rate can, in part, be attributed to a lack of diagnostic methods that enable the early detection of liver cancer. Hence, there is a need for further understanding of tumor biology and host response mechanisms so that new diagnostic and therapeutic tools can be developed. There has been a growing interest in using serum anti-tumor-associated antigen (TAA) antibodies as serological cancer biomarkers. This interest stems from the notion that these anti-TAA antibodies are 'sensors' or 'reporters' of molecular events associated with tumorigenesis. The persistence and stability of autoantibodies in the serum of cancer patients is an advantage over other potential markers, including the TAAs themselves, some of which are released by tumors but rapidly degrade or are cleared after circulating in the serum for a limited time. Furthermore, the widespread availability of methods and reagents to detect serum autoantibodies facilitates their characterization in cancer patients and assay development. The hypothesis is that 'customized' TAA arrays constitute promising and powerful tools for enhancing the serological anti-TAA antibody detection of cancer. The present review will focus on the recent advances in our studies primarily associated with the idea and possibility that autoantibodies to TAAs can be used as biomarkers in immunodiagnosis of HCC and other solid tumors.

The identification and characterization of tumor-associated antigens (TAAs) and their use in antigen mini-arrays for cancer immunodiagnosis has been of interest recently as an approach to cancer detection. In this study, autoantibodies in sera from a patient with HCC were used as probes to immunoscreen a HepG2 cDNA expression library for the identification of TAAs involved in malignant liver transformation. Recombinant proteins from two genes identified in this manner, Sui1 and RalA were expressed, purified and used as antigens in immunoassays to detect the presence of antibodies in sera from 77 patients with HCC, 30 with chronic hepatitis (CH), 30 with liver cirrhosis (LC) and 82 normal human sera (NHS). The prevalence of antibody to Sui1 and RalA in HCC were 11.7%(9/77) and 19.5%(15/77), respectively, which were significantly higher than prevalence in liver cirrhosis (3.3% and 3.3%), chronic hepatitis (0% and 0%) and normal human sera (0% and 0%). When Sui1 and RalA were added to a panel of eight other TAAs used in a previous study, the final cumulative prevalence of anti-TAA antibodies in HCC to the 10 TAA array was raised to 66.2%(51/77). The specificity for HCC compared with LC, CH and NHS, was 66.7%, 80.0%, and 87.8%, respectively. When anti-TAA was added to abnormal serum AFP as combined diagnostic markers, it raised the diagnostic sensitivity from 66.2% to 88.7%. AFP and anti-TAA were independent markers and the simultaneous use of these two markers significantly resulted in the increased sensitivity of HCC detection.

Many studies demonstrated that cancer sera contain antibodies which react with autologous cellular antigens generally known as tumor-associated antigens (TAAs). In our laboratories, the approach used in the identification of TAAs has involved initially examining the sera of cancer patients using extracts of tissue culture cells as source of antigens in Western blotting and by indirect immunofluorescence on whole cells. With these two techniques, we identify sera which have high-titer fluorescent staining or strong signals to cell extracts on Western blotting and subsequently use these sera as probes in immunoscreening cDNA expression libraries, and also in proteomic approaches to isolate and identify targeted antigens which might potentially be involved in malignant transformation. In this manner, several novel TAAs including HCC1, p62, p90, and others have been identified. In extension of these studies, we evaluate the sensitivity and specificity of different antigen-antibody systems as markers in cancer in order to develop "tumor-associated antigen array" systems for cancer diagnosis, cancer prediction, and for following the response of patients to treatment.

Although autoantibodies have been recognized as participants in pathogenesis of tissue injury, the collateral role of autoantibodies as reporters from the immune system identifying cellular participants in tumorigenesis has not been fully appreciated. The immune system appears to be capable of sensing aberrant structure, distribution, and function of certain cellular components involved in tumorigenesis and making autoantibody responses to the tumor-associated antigens (TAAs). Autoantibodies to TAAs can report malignant transformation before standard clinical studies and may be useful as early detection biomarkers. The autoantibody response also provides insights into factors related to how cellular components may be rendered immunogenic. As diagnostic biomarkers, specific TAA miniarrays for identifying autoantibody profiles could have sufficient sensitivity in differentiating between types of tumors. Such anti-TAA profiles could also be used to monitor response to therapy. The immune system of cancer patients reveals the immune interactive sites or the autoepitopes of participants in tumorigenesis, and this information should be used in the design of immunotherapy.

BACKGROUND/AIMS: Previous studies have demonstrated that during transition from chronic liver disease to hepatocellular carcinoma (HCC), autoantibodies can appear which are not detected in the prior pre-malignant conditions. These antibody responses may be stimulated by cellular proteins involved in carcinogenesis. This study determines the prevalence of antibodies to a selected panel of eight tumor-associated antigens (TAAs) in sera from patients with chronic hepatitis, liver cirrhosis and HCC, and considers the possibility and usefulness of antibodies to such a panel of TAAs in differentiating between these conditions. The panel of eight TAAs includes Imp1, p62, Koc, p53, c-myc, cyclin B1, survivin and p16 full-length recombinant proteins. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against eight selected TAAs in 30 sera from chronic hepatitis, 30 from liver cirrhosis, and 142 from HCC. Positive results were also confirmed by slot blot, Western blotting and immunoprecipitation assay. RESULTS: Antibody frequency to any individual TAA in HCC varied from 9.9% to 21.8%. With the successive addition of TAAs to a final total of eight antigens, there was a stepwise increase of positive antibody reactions reaching a frequency of 59.8% with whole cohort of HCC patients. This was significantly higher than the frequency of antibodies in chronic hepatitis (20%), liver cirrhosis (30%) and normal individuals (12.2%). CONCLUSIONS: This study demonstrates that malignant transition to HCC is associated with increased autoantibody responses to certain cellular proteins which might have a role in tumorigenesis, and shows that a mini-array of eight TAAs enhanced antibody detection for diagnosis of HCC. More studies in patients with HCC and precursor conditions such as chronic hepatitis, alcoholic hepatitis and liver cirrhosis using enlarged TAA mini-array panels might further improve the sensitivity and specificity of this mode of cancer immunodiagnosis. Its additional usefulness might be in the early detection of cancer in some patients with predisposing conditions.