Abstract In this paper we demonstrate that matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) mass spectrometry can be used to rapidly detect occult blood in human feces. The water-soluble hemoglobins, which are the most predominant proteins found in blood, were used as protein biomarkers for the presence of occult blood. The proteins were best extracted from feces using a small amount of pure water and then separated from the feces through centrifugation. We found that saturated 4-hydroxy-£\-cyanocinnamic acid (£\-CHC) solution was the best MALDI matrix for hemoglobin analysis; this matrix was mixed with the aqueous solution extracted from the feces at a volumetric ratio of 1:1. Although the MALDI-TOF MS results obtained using real samples were consistent with those provided through conventional chemical analysis, the detection limit of occult blood (0.1¡V1 £gg/mg of feces) in feces when using MALDI MS was approximately 10¡V100 times lower than that of the conventional approach. In addition, materials that commonly interfere with the conventional chemical assaying of occult blood in feces¡Xsuch as foods containing animal blood¡Xdo not interfere with the detection of the hemoglobin biomarkers during MALDI-TOF MS analysis.

Chapter 1. Deciphering the toxicity of Helicobacter pylori clinical isolates from gastric diseases patients using MALDI-TOF/MS and multivariate analysis. Helicobacter pylori (H. pyloyi) infection is associated with gastric diseases such as gastric polyp, chronic gastritis, gastric ulcer, gastric cancer, etc. In fact, most of the people infected not have the symptoms of gastric diseases due to the high degree of variability of gene with H. pyloyi and the specific immune responses of the hosts. In order to investigate the relationship between H.pylori and gastric diseases, the clinical strains of H. pylori isolated from patients from nine gastric diseases were extracted from the optimized extraction and analysis by MALDI-TOF/MS, then the high reproducible spectra were combined with multivariate statistical analysis including Principal Component Analysis (PCA), Hierarchical Cluster Analysis (HCA), Discriminant Analysis (DA) . In the result of PCA, there is no specific potential marker to discriminate the clinical strains to nine gastric diseases. In the result of HCA, the strains from different gastric diseases were clustered together means they have the similarity of the protein and metabolite. In the result of DA, the strains from gastric and non-gastric cancer were discriminanted by the discriminant function composed of thirty-eight discriminant variables in the spectra. This discriminant function would be confirmed by other clinical strains isolated from gastric diseases patients in the future and then would help to predict the the similarity of the protein and metabolite of the strains isolated from the gastric diseases patients whether gastric cancer or not. Chapter 2. Biomarker discovery in the stools of gastric cancer patients using MALDI-TOF/MS. According to the statistics of Republic 100 years from the Department of Health, cancer was the first of the ten lesding to death. With the modern change of eatiog habbits, gastrointestinal cancer has increased steadily. Gastrointestinal cancer accompanied occult gastrointestinal bleeding, and it is commonly detected by the fecal occult blood test (FOB). FOB including Guaiac-based fecal occult-blood test and immunochemical tests. Guaiac-based fecal occult-blood tests make use of the pseudoperoxidase activity of heme, and the reagent turns blue after oxidation by oxidants or peroxidases in the presence of an oxygen donor such as hydrogen peroxide, so it would have the potential of false-positive result. Immunochemical tests, which use antibodies detect against human hemoglobin with great sensitivity, but the tests are limited by loss of hemoglobin antigenicity at room temperature and require processing in a laboratory. In order to decrease the false-positive of detecting heme and decreasing the cost of the detection against hemoglobin in stools, in the study, we ues the distill water to extract the heme (m/z 616) and hemoglobin in stools and analysis with the reflectron and linear mode of MALDI-TOF/MS. In this study, at first, we used the stimulated stomach acid decomposing the hemoglobin to release the heme, to stimulate the gastrointestinal bleeding. Second, we used the distill water to extract the hemoglobin in stools, and detected by the linear mode of MALDI-TOF/MS, and the detection limit of MALDI-TOF/MS against hemoglobin in stool was better than the immunochemical tests. Third, the same strategy was applied to fifty-nine patients (including nineteen esophageal cancer patients, twenty gastric cancer patients and colorectal cancer patients) stools to detect heme and hemoglobin by MALDI-TOF/MS and the results were compared with the fecal occult blood test. In the detection of heme, MALDI-TOF/MS had not detect heme, but the Guaiac-based fecal occult-blood test had detected, it would be that the stools had the oxidants (not heme) to react the reagent. In addition, MALDI-TOF/MS had detected heme, but the Guaiac-based fecal occult-blood test had no results, those cases would be catched up in the future. In the detection of hemoglobin, using immunochemical tests to be the reference index, MALDI-TOF/MS had the false-negative result might come from the complicated matrix effect of stools, so that the hemoglobin could not form the good crystalline with matrix CHCA. The false-positive results of MALDI-TOF/MS might come from the criteria of hemoglobin signal.

Colorectal cancer poses a serious threat to the health and well-being of individuals, especially those at high risk or over the age of 50. Gone undetected, colorectal cancer is often fatal, however, preventive screening greatly reduces the number of people who may develop this disease. The Centers for Disease Control and Prevention developed the Colorectal Cancer Screening Demonstration Program in 2005 to assess the feasibility of a national colorectal cancer screening program serving low-income and un- or underinsured populations. Qualitative case study data from the Colorectal Cancer Screening Demonstration Program evaluation were analyzed in order to examine the start-up experiences of two of the programs involved. Results from this multiple case study document program models and describe facilitators, challenges, and participant perception of the expected impact on screening behavior. Further research on program implementation is needed to understand how program models perform and impact behavior once screening begins.

A Cochrane review conducted a meta-analysis looking only at FOBT for colorectal cancer screening. This review, based on published and unpublished data from 5 controlled trials, demonstrated that 3-card home FOBT conferred a reduction in colorectal cancer mortality of 16% (relative risk [RR]=0.84; 95% confidence interval [CI], 0.77-0.92) and a number needed to screen of 1173 (95% CI, 741-2807) to prevent 1 death from colon cancer over a 10-year period. If adjusted for adherence to screening, the reduction in mortality increased to 23% (RR=0.77; 95% CI, 0.57-0.89). In addition, long-term follow up of one of the RCTs in the review showed a continued reduction in colorectal cancer mortality of 34% (RR=0.66; 95% CI, 0.54-0.81) in subjects adhering to the FOBT screening protocol over a 13-year interval. Overall mortality did not differ between the screened and unscreened groups. A systematic review performed for the US Preventive Services Task Force (USPSTF) incorporated more recent data on colorectal cancer screening including colonoscopy. This review reached similar conclusions as above. This review also looked at office FOBT performed after digital rectal exam. It is important to note that a single office FOBT has a lower sensitivity than 3-card home FOBT and its effectiveness for reducing colorectal cancer mortality was unknown at the time of the systematic review. A subsequent 2005 Veterans Affairs prospective cohort study found that the sensitivity for detecting advanced neoplasia was only 4.9% for digital FOBT, and negative results did not decrease the likelihood of advanced neoplasia. The USPSTF review did not find any screening trials of colonoscopy but analyzed data from the National Polyp Study and a case-control study to draw its conclusions. The review reported an odds ratio for colorectal cancer mortality for patients who had colonoscopy to be 0.43 (95% CI, 0.30-63). The USPSTF review also looked at the sensitivity and adverse effects of FOBT compared to colonoscopy. One-time 3-card home FOBT had a sensitivity of 30% to 40% for detecting cancer. The sensitivity of one-time colonoscopy was difficult to determine since it was the criterion standard examination, but it was estimated to be greater than 90%, with a risk of perforation of 1/2000. The USPSTF review found both screening strategies cost-effective (<$30,000 per additional life-year gained) compared to no screening. FOBT had a cost per life-year saved of $5691 to $17,805 compared with $9038 to $22,012 for colonoscopy performed every 10 years.

Includes bibliographical references. / xiii, 260 leaves ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Examins Fecal Occult Blood Test (FOBT) screening as a possible means of reducing mortality from colorectal cancer in Australia. Consists of an evaluation of a FOBT screening program in South Australia (in terms of numbers of cancers detected, accuracy of the test used, costs of the program and characteristics of participants) and surveys of the general population and of South Australian general practitioners, providing information on knowledge, attitudes and practices in relation to colorectal cancer and its prevention. / Thesis (Ph.D.)--University of Adelaide, Dept. of Community Medicine, 1995?

Thesis (M.S.)--West Virginia University, 2006. / Title from document title page. Document formatted into pages; contains ix, 98 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 92-98).