Background: Blood coagulation is an essential defense mechanism to prevent bleeding. Disorders in the coagulation system can be severe and blood tests measuring the blood’s ability to coagulate are important. Activated partial thromboplastin time (APTT) is a blood test that measures blood coagulation time. An abnormal prolonged APTT can both be associated with a bleeding tendency or a risk of thrombosis. Additional blood tests are needed to discover the cause of a prolonged APTT. One potential test is the APTT mixing study, which can separate samples with and without inhibitors. The aim of this project is to investigate how the cause of a prolonged APTT is evaluated today and to examine if it is possible to indicate the cause of a prolonged APTT using the APTT mixing study performed on routine samples. The goal is to be able to indicate the cause of a prolonged APTT immediately when is it first discovered. This will save time and help guide the physicians in their work with the patient. Methods: Retrospective data is used to examine how the cause of a prolonged APTT is evaluated today. Samples with known cause of prolonged APTT are used to establish a cut-off value for the APTT mixing study to indicate the cause of a prolonged APTT. The cut-off values are then tested using routine APTT samples. Pre-analytical variables relevant to APTT are also investigated. Results: Today, specialized departments request most special coagulation blood tests. The APTT mixing study can separate samples with and without inhibitors with 90% specificity and sensitivity using index of circulating anticoagulants cut-off value of 16.0. In regard to pre-analytical variables, the centrifugation force affects the plasma platelet count but not APTT and sample storage has an affect on APTT. Conclusion: The APTT mixing study can be implemented as an additional test to indicate the cause of a prolonged APTT on routine samples.

No / Lymphocyte responses from 208 individuals: 20 with melanoma, 34 with colon cancer, and 4 with lung cancer (58), 18 with suspected melanoma, 28 with polyposis, and 10 with COPD (56), and 94 healthy volunteers were examined. The natural logarithm of the Olive tail moment (OTM) was plotted for exposure to UVA through 5 different agar depths (100 cell measurements/depth) and analyzed using a repeated measures regression model. Responses of patients with cancer plateaued after treatment with different UVA intensities, but returned toward control values for healthy volunteers. For precancerous conditions and suspected cancers, intermediate responses occurred. ROC analysis of mean log OTMs, for cancers plus precancerous/suspect conditions vs. controls, cancer vs. precancerous/suspect conditions plus controls, and cancer vs. controls, gave areas under the curve of 0.87, 0.89, and 0.93, respectively (P<0.001). Optimization allowed test sensitivity or specificity to approach 100% with acceptable complementary measures. This modified comet assay could represent a stand-alone test or an adjunct to other investigative procedures for detecting cancer.

Equine gastric glandular mucosal ulceration can have a prevalence of 58%, yet its etiology is poorly understood. In man Helicobacter pylori is the most common cause of gastritis and peptic ulcer disease. Helicobacter is uniquely able to colonize the stomach, via the action of cytoplasmic urease. Different Helicobacter species have been isolated from many mammals but none has yet been cultured from the horse. Three tests used to identify human Helicobacter infection were applied to the horse. Test 1: PCR amplification of Helicobacter specific DNA, n=12. Test 2: the Pyloritek™ rapid urease test (RUT), n=15. Test 3: the 13C-urea blood test, n=8. Gastroscopy and antral biopsy was performed in all horses. All horses demonstrated the presence of Helicobacter specific gene material by PCR. Biopsy specimens from 7/15 horses were urease positive by RUT. Significant 13C enrichment of the body CO2 pool was found in all horses after intragastric administration 13C-urea (p<0.05). As Helicobacter is currently the only known gastric urease positive microorganism, the demonstration of this activity in horses positive by PCR strongly supports the presence of an equine gastric Helicobacter species. Variations of 13C-urea blood test were further examined and a single protocol was found to be most applicable. As the horse is a hind gut fermenter, the effect of cecal urease on the test was examined by laparoscopic intracecal administration of 13C-urea. Significant cecal urease activity was demonstrated however the timing of peak 13C enrichment may limit any effect on the gastric test to 90 minutes onwards. / Master of Science

This thesis examines the benefits and costs of screening for colorectal cancer in the context of an organised population screening programme. It uses microsimulation modelling to derive an optimally cost-effective screening protocol for various combinations of the available screening tests. ¶ First a mathematical model for the natural history of colorectal cancer is derived, based on analyses of Australian population and hospital-based cancer registries combined with data from published studies. Then a model for population based screening is derived based mainly on data from published screening studies, including the four major published randomised controlled trials of faecal occult blood test (FOBT) screening. These two models are used to simulate the application of a screening programme to the Australian population. The simulations are applied to a period of 40 years following 1990 (the study’s base year), with both costs and benefits discounted back to the base year at an annual rate of 3%.¶ The models are applied to simulating a population screening programme based on FOBT with a colonoscopy follow up of positive tests. This simulation suggests that the optimal application of such a programme would be to offer annual screening to people aged 50 to 84 years. Such a programme would lead to a cumulative fall in years of life lost to colorectal cancer (YLL) of 28.5% at a cost per year of life saved (YLS) of $8,987. These costs and benefits are consistent with those arising from other currently funded health interventions. They are also consistent with the cost per YLS which Australian governments appear willing to pay for health interventions when justified on the basis of cost-effectiveness. The fall in colorectal cancer deaths from this screening programme should be first detectable by a national monitoring system after around three years of screening. However the full benefits from screening would not be realised before around 30 years of screening.¶ These simulations are based on the standard guaiac FOBT, but the results suggest that significant cost-effective gains could be made by using the newer immunochemical FOBT. Further cost-effect gains could be made by offering sigmoidoscopy every five years in addition to annual FOBT.¶ The models are then applied to simulating population screening programmes using colonoscopy and sigmoidoscopy as primary screening tools. Offering colonoscopy every ten years to all people aged from 45 to 85 leads to an overall fall in cumulative YLL of 37.6%, at a cost of $15,585 per YLS. Offering sigmoidoscopy every three years to all people aged 40 to 85 leads to an overall fall in cumulative YLL of 29.1%, at a cost of $4,862 per YLS. Both of these cost and benefit results are also consistent with the cost per YLS which Australian governments appear willing to pay. The fall in deaths with colonoscopy screening would also be detectable after three years of screening but the fall with sigmoidoscopy screening would not be detectable until after six years of screening. Sigmoidoscopy would need around 35 years of screening to reach its potential gains while colonoscopy screening would not reach its full potential during the 40 year screening period.¶ Finally the models are applied to targeting people at higher risk of cancer. The results show that offering colonoscopy every five years to people at higher risk because of a family history of colorectal cancer is a cost-effective addition to the annual FOBT screening programme.¶ An earlier version of chapter two of this thesis has been published as Stevenson CE 1995. Statistical models for cancer screening. Statistical Methods in Medical Research; 4: 19–23.¶ An expanded version of chapter two, along with parts of chapter one, has been published as Stevenson CE 1998. Models of screening. In: Encyclopedia of Biostatistics. Armitage P, Colton T, eds. John Wiley and Sons Ltd, pp 3999–4022.

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.

In Australia, colorectal cancer (CRC) is the most common registrable cancer affecting both men and women, and the third most common cause of cancer deaths. Clinical data from randomised, controlled trials indicate that population-based screening utilising the faecal occult blood test (FOBT) can reduce mortality from this disease. However, high adherence rates with repeated testing are required to secure these outcomes. This study examines repeat adherence with FOBT screening in a rural community two years after a first screening round was conducted. Patients, aged 50 to 74 years, registered with four local general practices were mailed a FOBT kit with a letter of invitation from their general practitioner. Following the intervention, 119 telephone interviews were conducted with adherers and non-adherers to examine knowledge and attitudes related to screening. Compliance with screening was recorded and compared with first round-data. Participation in the screening program was modest. Of the 3,406 participants eligible for both screening rounds, 34.1% and 34.7% participated in rounds 1 and 2, respectively. A majority of participants (56.8%) did not adhere to either screening, a quarter (25.7%) participated in both rounds, and 17.5% participated in one of the two rounds. First-round adherence was the strongest predictor of second-round adherence (OR=16.29; 95% CI: 13.58, 19.53) with 75.2% of first-round adherers completing a FOBT in round 2. Females were also more likely to adhere in both rounds, although the difference between females and males decreased across rounds. Knowledge and attitudes differed between adherers and non-adherers and are discussed within the context of the major findings. Results from this trial indicate that achieving high levels of compliance in a national screening program will be challenging. Strategies to increase repeat adherence are suggested.

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.

Helicobacter pylori (H. pylori) is a Gram-negative bacterium that infects and resides in the gastric mucosa of humans. Without treatment, H. pylori infection may cause chronic inflammation of the gastric mucosa. This inflammation creates progressive damage to the lining of the stomach and can lead to multiple diseases located in the upper gastrointestinal region. Worldwide prevalence of H. pylori infection is estimated to be close to 50%. H. pylori has been identified as the primary cause of peptic ulcer disease, gastric cancer, and mucosa-associated lymphoid tissue lymphoma. The purpose of this study was to identify the prevalence and risk factors associated with H. pylori infection among students, faculty, and staff at the University of Central Florida. A cross-sectional design with a convenience sample was implemented to acquire a study population of 60 participants. The sample was analyzed through the use of a twenty question survey and a rapid blood antibody test kit. The infection rate of the sample was 1.75%. Statistically significant results were found for the relationship between age and upper gastrointestinal symptoms. Trends were also noticed between alcohol consumption, stress levels, and upper gastrointestinal symptoms.

Background: Currently, participation in the NHS Bowel Cancer Screening Programme (NHSBCSP) is poor, with around half of all people invited returning their (FOBT) kits. The research programme aimed to investigate whether a general practitioner’s (GP) letter encouraging participation and a detailed leaflet explaining how to complete the (FOBT) included with the invitation materials would improve uptake. Methods: The research programme was divided into three phases which were designed to sequentially develop and evaluate the two interventions. The initial and second phases developed and refined the two interventions and the trial outcome measures with previous participants and stakeholder representatives. The final phase was a randomised 2x2 factorial trial conducted with people invited to screening in October 2009. Participants were randomised to either a GP’s endorsement letter and/or a detailed procedural leaflet with their FOBT kit. The primary outcome was verified participation in the NHSBCSP. Questionnaires were also used to evaluate participant perceptions of CRC screening and GPs views on involvement with the NHSBCSP. Results: The factorial trial demonstrated both the GP’s endorsement letter and the detailed procedural leaflet increased participation in the NHSBCSP. In the intention-to-treat analysis, participation improved by 6% for the detailed procedural leaflet and 5.8% for the GP endorsement letter 20 weeks after receipt of the FOBT kit. The random effects logistic regression model confirmed that there was no important interaction between the two interventions, and estimated an adjusted rate ratio of 1.11 (P=0.038) for the GP’s letter and 1.12 (P=0.029) for the leaflet. The per protocol analysis indicated that the insertion of an electronic GP’s signature on the endorsement letter was associated with increased participation (P=0.039). Conclusions: Including both an endorsement letter from each patient’s GP and a detailed procedural leaflet could increase participation in the NHSBCSP by around 10%, a relative improvement of 20% on the current participation rate. Both interventions were well-received by participants and there was minimal impact on GP workload.