ORAL PRESENTATIONS Development of a Rapid and Cost-Effective Method Suitable for Quantification of Global LINE-! Repetitive Element Methylation. Presenting Author- Sherry Taylor, Horizon Health Network Other Authors Will King , Queen’s University Stephen Pang, Queen’s University Nora Zwingerman, Queen’s University Janet Ashbury, Queen’s University Yat Tse, Queen’s University Background: Epigenetics refers to the study of heritable cellular changes that leave the DNA nucleotide sequence intact. Increasingly it is recognized that epigenetic mechanisms play an important role in cancer aetiology through alterations in gene activation, chromosomal stability, and genomic imprinting. The methylation of DNA is recognized as a key epigenetic mechanism in the regulation of genomic stability and cell growth, and evidence for its role in the development of a wide variety of cancers is accumulating. Many studies have demonstrated that the methylation status of the dispersed repetitive element LINE-1 closely parallels overall global methylation levels, indicating that LINE-1 methylation levels represent a valid surrogate measure of genome-wide methylation. Many of the methods used to measure LINE-1 methylation are technically challenging and labour intensive. The objective of the current investigation was to design a rapid, reproducible and cost-effective method to estimate overall LINE-1 methylation that is suitable for a large population-based study. Methods and Results: Using high resolution melt (HRM) curve analysis technology, a single intube assay was developed that is both linear (r > 0.9986) and has a high amplification efficiency (90-105%), capable of discriminating between patient samples with small differences in methylation, and suitable for quantifying a wide range of LINE-1 methylation levels (0 – 100%) including the biologically relevant range of 50 – 90% expected in human DNA. We have optimized this procedure to perform using 2 μg of starting DNA and 2 ng of bisulfite-converted DNA for each PCR reaction. Intra- and inter-assay coefficients of variation were 1.44% and 0.49%, respectively, supporting the high reproducibility and precision of this approach. Conclusions: In summary, we have developed a completely linear, quantitative HRM PCR method developed for the measurement of LINE-1 methylation. This cost-efficient, refined and reproducible assay can be performed using minimal amounts of starting DNA. These features make our assay suitable for automation and rapid high-throughput analysis of multiple samples from large population-based studies. HRM technology provides a cost- effective and flexible means to survey the methylation status of single or multiple sites of interest, including repetitive elements, to identify potential markers of risk in studies of cancer prediction, or as a companion to other genomic, proteomic or metabolomic strategies to further understand the role of methylation in the normal functioning of the genome or in disease processes. Saint John Hypertension Clinic: Efficacy and Barriers to Achieving Controlled Blood Pressure Presenting Author- Paul Bonnar, Resident, Horizon Health Network Other Authors Dr. Martin MacKinnon, Horizon Health Network Anita Fenn, Horizon Health Network Background: The importance of hypertension (HTN) is well established as a risk factor for many serious diseases such as coronary disease, ischemic stroke, intracerebral hemorrhage, heart failure, atrial fibrillation, and kidney disease. In 2007-2008, 29% of adults in the United States had HTN with control achieved in only 50%. Consequently, there must be barriers to improving the control of blood pressure (BP). HTN clinics provide an excellent population to study the outcomes of interventions and barriers to controlled BP. There were two objectives of this study: 1) determine the efficacy of the Saint John Regional Hospital (SJRH) HTN clinic and 2) identify factors that make HTN difficult to control by comparing patient characteristics among controlled and uncontrolled patients. Methods: This study was a retrospective analysis using a database compiled from chart reviews and questionnaires. The proportion of patients controlled at the initial visit was compared to the proportion at discharge. Patient characteristics were compared among patients who achieved controlled BP with those who did not achieve control at time of discharge. The characteristics included age, gender, renal disease or diabetes, initial number of medications, and number of visits. Results: The proportion of controlled patients significantly increased from the initial visit to discharge (initial = 44%; discharge = 73%; p = 0.002). At discharge, 15 patients had uncontrolled (unc) BP and 40 patients had controlled (con) BP. The systolic BP at the initial assessment did not differ between these two groups (unc = 155 ± 27 mmHg; con = 141 ± 25 mmHg; p = 0.068). By definition, the systolic BP at discharge was significantly lower in the controlled group (unc = 151 ± 16 mmHg; con = 121 ± 10 mmHg; p < 0.001). There were no significant differences among the characteristics of these two groups: age (unc = 53 ± 21; con = 57 ± 14; p = 0.828), percentage of males (unc = 40%; con = 48%; p = 0.619), percentage of kidney disease or diabetes (unc = 47%; con = 40%; p = 0.655), initial number of medications (unc = 3.4 ± 1.5; con = 3.0 ± 1.3; p = 0.276), and number of visits (unc = 3.1 ± 1.8; con = 3.0 ± 1.2; p=0.936). Discussion: This study shows that the SJRH HTN clinic improves BP control. However, no significant differences in patient characteristics were found among patients who achieved controlled BP and those who did not achieve control. Future research will include analysis of data after a determined number of visits to the clinic. Additionally, other patient characteristics can be included in the analysis such as race, tobacco use, diuretic use, and body mass index. “Doing what comes natural”: Understanding the direct caregivers’ experience of caring for residents with dementia Presenting Author- Sandee Hicks-Moore, UNB Saint John Background: Caring for persons with dementia living in long-term care is fraught with multiple challenges, yet a clear understanding from the unique perspective of the direct hands-on caregivers is missing from the literature. Focusing on the perspectives of the direct caregivers, namely the personal support workers (PSWs) and licensed practical nurses (LPNs), this research explores their understandings of dementia care and the realities that they experience on a day-today basis working in long-term care. Method: The research design is informed by hermeneutic phenomenology and seeks to answer the following two questions: 1) What meanings do direct caregivers -- PSWs and LPNs -- in long term care develop about their experiences of caring for persons with dementia? 2) How may these meanings change, if at all? The stories, insights and experiences of 14 direct caregivers caring for persons with dementia and working in five different long-term care (LTC) facilities located in rural and urban settings in southern New Brunswick will be highlighted. Results: Three themes emerged from the analysis of the data.1) Adaptation refers to the caregivers’ abilities to adjust to the unpredictable and often psychological and physically overwhelming situations they experience in caring for residents with dementia. These caregivers have learned how to rapidly adapt and temper their care to meet the multiple and often competing demands of the institution and the residents. 2) Inclusion refers to how the caregivers seek ways to stabilize themselves and gain acceptance in the workplace. To survive, the caregivers rapidly learn to feel included, but on their own terms. They assess the context of their workplace and establish connections with others in ways that help them maintain psychological stability. Such connections with peers, co-workers, or residents foster a sense of belonging, a sense of purpose and gives meaning to their work. 3) Denied professionalism refers to the lack of credit given to their knowledge and skills by co-workers, supervisors and administrators of higher status. Discussion: This presentation will highlight the findings of this research study in an effort to broaden our understanding of the experiences of the direct caregivers caring for persons with dementia. In honouring the work of the direct caregivers, this research will illuminate their perceptions and challenges. The knowledge gained from this study may be useful in enhancing educational and support programs for PSWs and LPNs. Optimization of the microplate resazurin assay as a screening tool for natural products with anti-tuberculosis activity Presenting Author- Taryn O’Neill, UNB Saint John Other Authors Duncan Webster, Horizon Health Network John A Johnson, UNB Saint John Christopher A Gray, UNB Saint John Background: Natural products chemistry has provided novel bioactive compounds for drug discovery through bioassay-guided fractionation techniques. The microplate resazurin assay (MRA) is used for the isolation of anti-mycobacterial compounds from phytochemical extracts. However, we found that analysis of methanolic extracts using the MRA problematic, providing inconsistent results. In this study we optimize the efficiency of the MRA for both extract screening and bioassay-guided fractionation of phytochemical extracts. Methods: MRA aspects evaluated include: DMSO effect on mycobacterial growth; optimization of resazurin indicator concentration; plate type used; and assay duration. All tests were conducted on avirulent M. tuberculosis strain H37Ra as it is more safe and accessible and has been demonstrated to be an acceptable surrogate strain for H37Rv. Results: The optimal DMSO solvent concentration is 2 % as test samples were fully dissolved with minimal impact on mycobacterial growth. The ideal resazurin indicator concentration was 0.125 mg/mL in 10% Tween 80 (1:1) delivering optimal fluorescence and colourimetric readings. Black plates delivered optimal results when read fluorometrically and clear plates when observing colour change. Traditionally, the MRA is a 7-day assay, however, may be completed in 2 days. The assay was verified using methanolic extracts of Heracleum maximum. Through bioassayguided fractionation using the optimized MRA, the natural product falcarindiol was identified as the primary constituent conferring anti-mycobacterial activity in extracts of H. maximum. Discussion: We have optimized the MRA to deliver consistent results more rapidly than the standard 7-day protocol thus increasing the efficiency of both extract screening and bioassayguided fractionation. A Feasibility Study of a Health Intervention for Women Who Have Experienced Intimate Partner Violence Presenting Author- Judith Wuest, Horizon Health Network Other Authors Marilyn Merritt-Gray HRP, UNB Nursing Faculty Norma Dubé, ADM, Government of New Brunswick, Women’s Issues Marilyn Hodgins, Assoc. Prof, UNB Nursing Faculty Jo Ann Majerovich, Physician, UNB Student Health Center Marilyn Ford-Gilboe Prof, University of Western Ontario, School of Nursing Colleen Varcoe, Prof, University of British Columbia, School of Nursing. Background: Intimate partner violence (IPV) is a major public health problem affecting 1 in 3 Canadian women. IPV has serious negative effects on the physical and mental health of women that persist long after the crisis of leaving. In our previous research, we found that Canadian women separated from abusive partners an average of 20 months have poorer physical and mental health and greater use of health services than Canadian women in general. We estimated annual costs, excluding medication, to the health care system attributable to violence to be almost $5000 per woman. Importantly most of these health problems are amenable to treatment and help is essential to prevent them from becoming worse. Yet, little attention has been given to the development and evaluation of health interventions for abuse survivors after leaving. Based on these findings and our qualitative theory of strengthening capacity to limit intrusion, we developed a 6 month, community-based primary health care intervention called iHEAL (Intervention for Health Enhancement After Leaving) to address the effects of gender-based violence on women’s health and quality of life. For complex community-based interventions, exploratory studies are necessary to determine effectiveness and feasibility in different contexts. Methods: We partnered with New Brunswick Women’s Issues Branch and Department of Health, and with Liberty Lane Inc. to obtain a Partnerships in Health System Improvement grant from CIHR and NBHRF to fund this mixed-method intervention feasibility study consisting of repeated measures (pre-intervention, post intervention at 6 and 12 months) and qualitative description of the study processes. Our main hypothesis was that women’s quality of life (QOL measure) and physical and mental health (SF-12) would improve post-intervention. The iHEAL is being delivered in 12-14 individual sessions by nurses and existing domestic violence outreach workers in Fredericton, Miramichi, Saint John and Sussex. We recruited 52 survivors who had separated from their abusive partners in the past 3 years. Qualitative interviews were conducted with participants and interventionists to garner their experiences with the iHEAL. Process notes were kept of bi-weekly interventionist meetings and training sessions. Chart reviews have provided data on the number and duration of contacts, time spent on each component, referrals, and outcomes. Post-intervention (6 months) data will be complete in February 2012. Results: Based on preliminary analysis, using paired t-tests for 37 women who have completed the 6 month post-intervention data collection, our hypotheses have been partially supported. Quality of life and mental health significantly improved; physical health did not. As well, mastery, and social support significantly improved; symptoms of depression and posttraumatic stress disorder, chronic pain grade and disability from chronic pain significantly declined. No significant differences were found for chronic pain intensity or social conflict. Qualitative analysis is ongoing to provide a contextual assessment of the feasibility and acceptability of the iHEAL. Discussion: We will discuss the preliminary implications of our 6 month post-interventions findings for programs and policy focused on improving the health of women trauma survivors, particularly clinical and cost effectiveness of health and social service interventions. Bioassay screening of traditionally used medicinal plants and the isolation of apoptosis inducing compounds. Presenting Author- Caitlyn Carpenter, Student, UNB Saint John Other Authors Dr. Christopher Gray, UNB Saint John Dr. Gilles Robichaud, University of Moncton Background: An abundance of new chemical structures with diverse biological activities can be found through natural products research (Harvey et al. 2010). Approximately half of the drugs that are in clinical use today can be traced back to either a natural product source or are based on a natural product template (Paterson and Anderson 2005). Ethnopharmacology is a useful alternative to random screening as it uses traditional knowledge to focuses on plants that have been used for their medicinal properties and are therefore more likely to possess biological activity (Gertsch 2009). The link between ethnopharmacology and anti-cancer drug discovery has been confirmed through many different studies (Heinrich and Bremner 2006). Such research has validated that many medicinal plants do have healing properties and that ethnopharmacology plays an extremely important role in discovering new natural products that possess the potential to become new drugs. As such, one of the main goals of the UNB Natural Products Research Group is the isolation and identification of new cancer drug candidates from traditionally used medicinal plants. Methods: Twelve plants were identified through their previous use as traditional medicines by the indigenous peoples of Canada, and their availability in New Brunswick. Extracts of the plants were tested for bioactivity against breast cancer cell lines (e.g. MDA231) to evaluate their anti-proliferation and apoptosis activity. The compounds responsible for the activity were then isolated by bioassay guided fractionation and identified from their spectroscopic and spectrometric data. Results: The preliminary screening of the twelve plants indicated Juniperus communis was a good candidate species for further investigation. The apoptosis inducing constituents of the juniper extract were isolated using various separation techniques and identified as isocupressic acid and deoxypodophyllotoxin through detailed analysis of 1D and 2D nuclear magnetic resonance spectra and mass spectrometric data. Discussion: Our research has provided further validation of the potential of traditionally used medicinal plants to be a source of new drug candidates. Current and future work includes: further testing on isocupressic acid to better understand the mode of action the apoptosis induction caused by this compound; investigating whether the bioactive compounds are produced by Juniper communis de novo, or rather by an endophyte found within the plant tissues. Therapeutic Commitment and Care of Persons with Mental Illness: A Survey of Nurse Practitioners’ Role Perceptions Presenting Author- Anne Marie Creamer, Horizon Health Network Other Authors- Dr. Wendy Austin RN PhD,University of Alberta, Faculty of Nursing and the Dossetor Health Ethics Centre Dr. Judy Mill RN PhD,University of Alberta, Director, PAHO/ WHO Collaborating Centre for Nursing & Mental Health, Faculty of Nursing Dr. Beverley O’Brien RN DNSc, University of Alberta, Faculty of Nursing Background: Despite tremendous potential for Nurse Practitioners (NPs) to address gaps in health care for individuals with mental illness, little is known of how competent, committed and supported they feel while working with this population. Objectives: The purpose of this research study was to determine how NPs rate their levels of therapeutic commitment, role competency and role support when working with persons with mental illness, including mental health problems. Method: The therapeutic commitment model was the theoretical framework for this study. A sample of 680 Canadian NPs who were accessed through two territorial and nine nursing associations completed a cross-sectional postal survey which was comprised of the Mental Health Problems Perceptions Questionnaire (MHPPQ), a demographic section and 3 open-ended questions. Measures of central tendency, Pearson’s correlational coefficient, and ANOVA were calculated. The results of the short answer questions were collated and common themes were identified Results: On a Likert – like scale of 1 (strongly disagree) to 7 (strongly agree), NPs reported mean scores of 5.05 (SD 0.83), 5.02 (SD 0.88) and 4.86 (SD 1.27) on the Therapeutic Commitment (TC), Role Competency (RC) and Role Support (RS) subscales respectively. The three subscales were correlated, with RC and TC being the most strongly associated (r = .754, p <.001). Scores for a composite variable, Confidence to Manage, which examined NPs’ confidence to manage 7 mental health disorders and suicide ideation were correlated with TC, RC and RS scores. Feeling ill-equipped to work with this population, knowledge of community mental health services, ratings of relevant theoretical and clinical NP education, previous mental health and NP work experience were all correlated with the 3 subscales. Differences in categories for population size of the community the NP worked in, frequency of collaboration for psychiatric reasons, time since the NP accessed mental health education and level of nursing education were found to impact levels of TC, RS, RC and Confidence to Manage. A significant number of NPs reported feeling inadequately prepared to manage the care of persons with mental illness and recommended either adding or increasing mental health education in their NP programs. Most NPs saw the care of this population as part of their role and wrote about the need for increased support for persons with mental health issues. Discussion: The results of this study found positive correlational relationships among the subscales of the therapeutic commitment model. Factors which impact TC, RC and RS were identified and provide guidance for policy and educational development. More study of factors which impact feelings of being supported in the role is needed. The model posits that higher levels of TC lead to improved patient outcomes; further research is needed to look at this. Percutaneous Thoracolumbar Stabilization For Trauma: Surgical Morbidity, Clinical Outcomes and revision surgery Presenting Author -Dr. Neil Manson, Canada East Spine Center, Horizon Health Network OTHER AUTHORS Renee Elliott, Canada East Spine Center Edward Abraham, Canada East Spine Center, Horizon Health Network Background: Percutaneous pedicle screw-rod instrumentation (PercStab) without direct decompression or fusion is a surgical option to manage thoracolumbar trauma. The current standard of care includes instrumentation removal following osseoligamentous healing. It is hypothesized that instrumentation removal is not required following PercStab. The purpose of the present study is to evaluate the utility of PercStab in the following areas: 1) patient satisfaction, 2) return of function 3) need for repeat surgery including instrumentation removal or revision decompression and/or fusion procedures. Methods: A Retrospective review of a prospective database identified patients receiving PercStab for trauma from January 2007 to August 2011. These fractures fulfilled the Spine Trauma Study Group criteria to be classified as unstable. Validated clinical outcome measures, patient demographics, perioperative data, and the need for further surgery were assessed via clinic follow up, chart review, and telephone interview. Medians and ranges reported. Outcome measures intra-op consisted of OR time / level, estimated blood loss / level and intraoperative complications. Post-op complications and length of surgery were analyzed in addition to VAS back and leg pain scores, Oswestry Disability Index (ODI), a Likert-type questionnaire and return to work (RTW) and normal activities. Results: Twenty-six trauma patients with a median Injury Severity Score 10 (9 – 41) received PercStab to treat spinal instability over 2 levels (range 1-5) and were followed for 22 months (2 months - 4.5 years). Minimal surgical morbidity was incurred: OR time: 36.5 (25 – 63.5) minutes/level, blood loss/level: 40 (12.5 – 250) cc, time post op to hospital discharge: 6 (1 – 37) days. Patients reported satisfaction: VAS back: 2 (0 – 8), VAS leg: 1 (0 – 7), ODI: 16 (0 – 54), 3 (2 weeks – 6) months to return to work or regular activities. Patients scored a median outcome satisfaction of 5 (3.75 – 5) out of 5 on a likert-type questionnaire. Only four patients required instrumentation removal: two for screw loosening causing back pain and two thin patients for screw prominence causing discomfort with direct pressure. The remaining 22 / 26 (85%) of patients had no symptoms or complications related to the implants and there was no need for revision surgery for post-traumatic deformity, pseudarthrosis or infection. Discussion: This surgical option provides rapid mobilization and discharge from hospital, medium-term satisfaction, with minimal surgical morbidity. The majority pf patients returned to work and normal activities on average three months post-op. Instrumentation removal can be considered on an individual basis. Further research is required to quantify the utility of this technique in comparison to traditional surgical options. St Stephen High School Healthy Student Body Initiative Presenting Author- Sohrab Lutchmedial, NB Heart Centre, Horizon Health Network Other Authors Yvonne Bartlett, Charlotte County Hospital, Horizon Health Network Amy Watling, NB Heart Centre Research Initiative Denise LeBlanc-Duchin, Research Services, Horizon Health Network Don Walker, St Stephen High School Dr. Ansar Hassan, NB Heart Centre, Horizon Health Network Background: An alarming number of youth today are faced with the early stages of chronic health issues such as obesity, hypertension, and high cholesterol levels. Given the link between health behaviours initiated in childhood and chronic health disease risk later in life, St Stephen High School personnel and local health care providers expressed concern about activity levels, risky lifestyle choices, and nutritional habits of their student population. The St Stephen High School Healthy Body Initiative started in November 2010 with 145 grade 10 students receiving health report cards. AstraZeneca Canada provided funding to continue the initiative for the new cohort of grade 10 students in November 2011. The initiative is aimed at improving the health and overall well-being of students. Methods: Parents received an information letter and consent form to permit their children to participate in the screening day. 140 Grade 10 students were participated in the screening in November 2011. Height, weight and waist circumference were recorded by nurse volunteers from the community. Blood pressures were recorded by manual cuff measurements using appropriately sized equipment. LDL, HDL, triglycerides and blood glucose were measured using a finger prick point of care device supplied by AstraZeneca. Body Mass Indexes (BMIs) were calculated from the height and weight obtained. Students also completed a Healthy Lifestyle Questionnaire, which assessed their nutrition, lifestyle, and physical activity habits, and a SelfEsteem Questionnaire. All information was provided to students and parents on a health report card. Results: 68 male and 72 female students participated in the 2011 Health Screening Day. Mean BMI for 2011 was 24.14, which decreased from 2010 (24.94). Self-esteem scores ranged from 3 to 30, with a mean of 19.34 in 2011. Mean self-esteem of the 2010 students was 20.04. Only 15% of students stated they are getting the 60 minutes of exercise per day that is recommended for youth, but 20% stated they are playing 4 or more hours of video games per day. Further data analysis will be completed in the coming months. Discussions: At-risk students (defined as students with three or more risk factors for heart disease) will meet with a health care provider to discuss the results of their report card and determine what plan of care they would like to follow. They will be given the option of attending one-on-one or group educational sessions with a certified dietician, and working with a personal trainer for six to eight weeks in the spring of 2012. According to the Public Health Agency of Canada (2008): “research consistently demonstrates that health and education are inextricably linked – and the most effective way to address issues such as maintaining a healthy weight, is through a comprehensive school health approach” (2). Ultimately, the adoption of a school based healthy lifestyle program is intended to have a positive effect on student health, well-being and educational outcomes. Anti-TB natural products from Aralia nudicaulis Presenting Author- Li Haoxin, UNB Saint John Other Authors Dr. Duncan Webster, Horizon Heath Network Dr. Chris Gray, UNB Saint John Dr. John Johnson, UNB Saint John Background: Aralia nudicaulis L., wild sarsaparilla, is a member of the Araliaceae or ginseng family that is indigenous to North America and was used extensively as a traditional herbal medicine by the First Nation communities of Canada. The Mi’kmaq and Maliseet First Nation peoples use Aralia rhizomes to prepare medicines for the treatment of wounds and respiratory ailments such as coughs. Results of a previous study showed that the aqueous extract of A. nudicaulis rhizomes possessed moderate anti-mycobacterial activity. The objective of the present research was to isolate and identify the anti-mycobacterial constituents of A. nudicaulis. Methods: The micro plate resazurin assay was used to assess the anti-mycobacterial activity of methanolic extracts of A. nudicaulis against Mycobacterium tuberculosis (H37Ra) and facilitated the isolation of active compounds through bioassay guided fractionation. Extracts of A. nudicaulis were fractionated by liquid-liquid partitions, column chromatography and high performance liquid chromatography (HPLC). The structures of the active compounds were elucidated through analysis of one dimensional and two dimensional nuclear magnetic resonance (NMR) data, mass spectrometry and polarimetry. Results: Two C17 diynes were found to be responsible for the anti-mycobacterial activity of A. nudicaulis rhizomes. The diynes were identified as falcarinol and panaxydol with MICs of 25.6 µM and 36.0µM and IC50 of 15.3 µM and 23.5µM against M. tuberculosis (H37Ra), respectively. Discussion: The isolation of two natural products with significant anti-mycobacterial activity from the rhizomes of A. nudicaulis has validated one of the traditional medicinal uses of this plant by Canadian First Nations. This was also the first report of anti-TB activity of the C17 diyne panaxydol. Evaluation of the low-threshold/high-tolerance methadone clinic in Saint John, New Brunswick, Canada - August 2009 to Present. Presenting Author- Tim Christie, Director, Ethics Services, Horizon Health Network Objective: To determine whether the low-threshold and high-tolerance (LTHT) aspects of this methadone maintenance treatment clinic (MMT) have had negative repercussions on patient outcomes by reporting the one-year retention rate, the prevalence of illicit opioid and cocaine use, and the costs of this clinic. Methods: The one-year retention rate was determined by collecting data on patients between August 04, 2009 and August 04, 2010. The number of patients enrolled in the clinic during this period was compared to the number of patients still in the program after receiving 12-months of treatment. The prevalence of illicit opioid and cocaine use was determined using a randomly selected retrospective cohort of patients. For each patient the results of six consecutive urine tests for the most recent three months were compared to the results of the first six consecutive urine tests after program entry. The costs were determined by delineating the cost of methadone hydrochloride, laboratory services, operational costs and pharmacy costs. Results: The one-year retention rate was 95%, 67% of the cohort achieved complete abstinence from illicit opioids and an additional 13% abstained from cocaine use after stabilizing on methadone. The cost of treating a patient is $5,882.25/year. Conclusion: Neither the low-threshold or high-tolerance aspects have had any negative impacts on patient outcomes. In terms of the “low-threshold” aspect, eliminating the intake procedures and team assessments do not appear to have had any negative effects on outcomes. Regarding the “high-tolerance” aspect there are no observed negative consequences associated with the absence of group or individual counselling, scheduled (as opposed to random) urine tests, and the “noinvoluntary-discharge” policy. The immediate acceptance of this service demonstrates that it could be an acceptable alternative to the current waitlist strategy in New Brunswick. Optimization of a molecular genetic assay for the sensitive detection of KRAS mutations in colorectal carcinoma Presenting Author- Isaac Miao, Student, Dalhousie Medicine New Brunswick Other Authors Dr. Sherry Taylor, Molecular Diagnostics/Laboratory Medicine, Horizon Health Network Background; Activating mutations of codons 12 and 13 of the KRAS gene occur frequently in colorectal carcinomas. Clinical trials have shown patients with metastatic colorectal cancer benefit from treatment using monoclonal antibodies such as Vectibix (panitumumab) to block EGFR overexpression, as long as point mutations in codons 12 or 13 of the KRAS gene are absent. As patients with mutations in their KRAS gene do not respond positively to Vectibix therapy, Health Canada has recently required use of a companion diagnostic kit to maximize health care resources. Since being approved by Health Canada, the DxS KRAS mutation kit has doubled in price to $200-$500 per patient tested, making it less attractive for use by clinical laboratories. Alternatives to the diagnostic kit were pursued in an attempt to minimize costs and turnaround time. Methods: The commercial DxS (Amplification Refractory) kit and a less costly method, SNaPshot (Primer Extension), were compared in terms of cost, turnaround time and KRAS mutation sensitivity using 25 cases of colorectal cancer previously analyzed by an external laboratory. Two extraction methods, Qiagen FFPE and PicoPure were also compared in terms of cost, turnaround time and DNA quality. Results: The Qiagen FFPE kit was consistently found to produce superior results in DxS runs with the same turnaround time of 16 hours despite being twice as costly at $30 per sample. KRAS mutation status was obtained for all samples assayed and were compared with results from the external laboratory. Both DxS and SNaPshot met the required analytic sensitivity of 1% mutant to wildtype sequence. One sample was discordant by both assays with those previously reported due either to the assays or sampling from a different location in the tumour tissue than originally performed. Both methods were also able to detect a mutation in a sample that the external laboratory had considered "insufficient DNA". SNaPshot detected the Gly13Asp variant as an artifact in addition to the correct genotype in four samples. Cost of analysis using SNaPshot was found to be $68 with a turnaround time of 8 hours, significantly lower than the DxS price of $200-500 and the external laboratory turnaround time of 3+ weeks. Discussion: Due to the inefficiencies of the current method of sending low amounts of DNA samples to an external lab, alternatives were sought out with the goal of reducing costs and turnaround time. Methods such as COLD-PCR with High Resolution Melt were deemed to be unacceptable due to the need for a large sample number to be tested in order for the software to establish a baseline The SNaPshot technique was found to be 95% concordant with the external lab results as sensitive as the DxS commercial kit, both of which detected mutations below the 1% level. DNA quality from the fixed tissues as well as non-specificity of primers is being investigated as the cause of the Gly13Asp artifact. Further validation and troubleshooting of the SnapShot method is required before it can be adopted for clinical use.