2017 CNMSRS COMPENDIUM by Kimberley Ormiston

Table of Contents 1 Skyline from The Forks Credit: Dan Harper

WELCOME

ORAL PRESENTATION CHAIRS AND ORGANIZING COMMITTEE

AGENDA

POSTER POSITIONS AND ORAL PRESENTATION SCHEDULE

DETAILED SCIENTIFIC SCHEDULE

A.M. SESSION: LIGHTNING ORAL PRESENTATIONS

Nathan Corbett – Dalhousie University

Matthew Danker – McGill University

Kayla Furlong – Memorial University

Jeffrey Grab – University of Alberta

Jack heard – University of Manitoba

Weining Yang – University of Toronto

Anna Kovalchuk – University of Calgary

POSTER DISCUSSION GROUP (by order of poster display #)

Emma Ali - Schulich School of Medicine & Dentistry 500

Alexei Berdnikov – University of Manitoba 501

Olivia Cook – University of Ottawa

502

Nathan Corbett – Dalhousie University Medical School 503

Matthew Dankner – McGill University

504

Victor De Broux-Leduc - Université de Montréal 505

Kayla Furlong – Mermorial University of Newfoundland 506

Louis Gervais – Université de Montréal 507

Jeffrey Grab – University of Alberta 508

36 2

Andrew Hanna – Dalhousie University Medical School 509

Irene Harmsen – University of Toronto

510

Jack Heard – University of Manitoba 511

Ryan Henrie – University of Manitoba 512

Nicholas Jette – University of Calgary 513

Cameron Kaye – University of Manitoba 513

Katrice Kazmerik – University of Manitoba 515

Noren Khamis – University of British Columbia 516

Anna Kovalchuk – University of Calgary 517

Alexander Levit – Schulich School of Medicine and Dentistry 518

Donna Liao – University of British Columbia 519

Amber MacLellan – Dalhousie University Medical School 520

Andrew McDermid - University of Manitoba 521

Ilya Mukovozov – University of Toronto 522

Mathieu Nadeau-Vallée - Université de Montréal 523

Alex Nantsios – University of Ottawa 524

Siddharth Nath – McMaster University 525

Vincent Picher-Martel – Laval University 526

Sheharyar Raza – University of Toronto 527

Quinlan Richert – University of Manitoba 528

Nadine Rockwood – Memorial University of Newfoundland 529

Joanna Ryan – Schulich School of Medicine and Dentistry 530

Nicholas Sajko – Dalhousie University Medical School 531

Alyssa Smith – McGill University 532

Naveet Sohi – University of Alberta 533

Xiao Jing (Lily) Sun – University of Alberta 534

Daniyil Svystonyuk – University of Calgary 535

Stephanie Totten – McGill University 536

Karim Wafa – Dalhousie University Medical School 537

Ali (Qinyuan) Xu – University of British Columbia 538

66 3

Weining Yang – University of Toronto 539

Russell Yanofsky – McGill University 540

Albert Yeung – University of Manitoba 541

Nadège Zanré - Université de Montréal 542

Kirill Zaslavsky – University of Toronto 543

Pegah Afshar – University of Manitoba 544

P.M. SESSION: LIGHTNING ORAL PRESENTATIONS

Kirill Zaslavsky – University of Toronto

Vincent Picher-Martel – Laval University

Naveet Sohi – University of Alberta

Daniyil Svystonyuk – University of Calgary

Stephanie Totten – McGill University

Ryan Henrie – University of Manitoba

Alis (Qinyuan) Xu – University of Manitoba

Nadège Zanré - University of Montreal

Ilya Mukovozov - University of Toronto

PARTICIPATING SCHOOLS

PARTICIPANT CONTACT INFORMATION

SPONSORING AGENCIES

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Welcome to the 9th Annual Canadian National Medical Student Research Symposium! The opportunity to indulge your curiosity, make a discovery and thereby make a change in the world is an incredible privilege that few have. Researchers are one group that is very fortunate to do so. At the same time, there is no doubt that learning how to conduct and interpret research, how to develop it into a substantial portion of your career, and how to maintain a successful program is hard work that is fraught with uncertainly, insecurity and the occasional burst of excitement. Over the next several days you have the opportunity to share your experiences, your emerging research projects and your own experimental findings with others. This is an excellent way of helping you determine if research truly is a passion for you. For those individuals who can combine the superb breadth of biological understanding that a medical degree provides with the ability to ask well focused, targeted questions that develops in a graduate degree program, there are a great many professional opportunities. Future options range from leading large multicentre research groups, to running your own laboratory, to being a key collaborator with a sophisticated understanding of the benefits (and limitations) of biomedical research. The trainees this year represent medical schools, and research disciplines, from across Canada. About one third of our 53 attendees are in their first year of hands-on research, one third have intermediate experience and another third are engaged in joint MD PhD programs. As the abstracts demonstrate, your work ranges from studies of individual gene polymorphisms or molecules through to population and health systems research. Because so much ground breaking research is highly multidisciplinary, we have designed the program to increase academic and social interactions between trainees such as yourselves with those enrolled in non-clinical graduate training programs. Make the most of the opportunity! I want to personally thank the individuals and organizations that have enabled us to offer you this special opportunity. Money is never is excess supply and they have gone to great effort to support this initiative. Contributors are listed at the back of the Symposium program. We welcome you and hope you enjoy the novel science, the professional development activities and the pleasures of meeting others with similar interests during your three days here.

Kent T. HayGlass PhD Canada Research Chair in Immune Regulation Director, Advanced Degrees in Medicine University of Manitoba

Session Chairs for Oral Presentations: A.M. Session – Karim Wafa – Dalhousie University Victor De Broux-Leduc - Université de Montréal

P.M. Session – Donna Liao – University of British Columbia Jeffrey Grab - University of Alberta

ORGANIZING COMMITTEE Dr. Kent HayGlass – Director, Advanced Degrees in Medicine, CNMSRS Coordinator Kimberley Ormiston – Program Coordinator, Graduate and Advanced Degrees Education Program Coordinator Cameron Kaye – Committee Member and MD/PhD student Andrew Denisuik – Committee Member and MD/PhD student Shenghua Zhu – Committee Member and MD/PhD student

Detailed Scientific Schedule 0845 – 9:55

A.M. Session – Lightning Oral Presentations Each presentation will be 6 minutes for presentation and 4 minutes for questions; 10 minutes in total. Procurity Lecture Theatre Main Floor – Apotex Centre Chairs: TBA Student

Poster #

School

Abstract Title

Nathan Corbett

503

Dalhousie University

Suicide risk correlates with lithium response in patients with bipolar disorder

Matthew Danker

504

Dalhousie University

Orthotopic Patient Derived Xenografts as Relevant Models of Brain Metastasis

Kayla Furlong

506

Memorial University

Body mass index-for-age and weight-forlength < 2 years are associated with weight and cardiometabolic outcomes midchildhood

Jeffrey Grab

508

University of Alberta

Robotic TMS mapping of motor cortex in the developing brain

Jack Heard

511

University of Manitoba

Gender Dysphoria Assessment and Action for Youth: Review of Healthcare Services and Experiences of Trans Youth in Manitoba

Weining Yang

539

University of Toronto

Alterations in cervical glutamatergic interneurons preserves respiratory function in cervical spondylotic myelopathy (CSM)

Anna Kovalchuk

517

University of Calgary

Chemotherapy, tumours, and the brain.

10:00– 11:45 – Poster Presentations

Coffee Available The Elevator Presentation: 2 minute oral summary of study rationale and a single key finding to start the conversation as people come by. Those who choose to do so, can ask you for details/have a discussion. Student

Poster #

Emma Ali

500

School

Abstract Title

Schulich School of Medicine and Dentistry

Institutional Practices And Predictors Of Survival For Limited-Stage Small Cell Lung Cancer – A Retrospective Analysis

Alexei Berdnikov

501

University of Manitoba

To examine the mechanisms through which adipose-derived stem cells affect healthy and malignant breast epithelial cell proliferation

Olivia Cook

502

University of Ottawa

Application and Usefulness of Outpatient Cardiac Testing Among Emergency Department Patients with Syncope

Nathan Corbett

503

Dalhousie University Medical School

Suicide risk correlates with lithium response in patients with bipolar disorder

Matthew Dankner

504

McGill University

Orthotopic Patient Derived Xenografts as Relevant Models of Brain Metastasis

Victor De BrouxLeduc

505

Université de Montréal

Espace Transition au Musée – Deconstructing stigma one painting at time

Kayla Furlong

506

Memorial University of Newfoundland

Body mass index-for-age and weight-forlength < 2 years are associated with weight and cardiometabolic outcomes midchildhood

Louis Gervais

507

Université de Montréal

Survival of ventral midbrain dopaminergic neurons during aging is not regulated by netrin receptor UNC5A

Jeffrey Grab

508

University of Alberta

Robotic TMS mapping of motor cortex in the developing brain

Andrew Hannah

509

Dalhousie University Medical School

Association between physical activity and cardiac risk factors in disease prevention within cardiac rehabilitation populations.

Irene Harmsen

510

University of Toronto

Use of magnetoencephalography to examine cortical responses to deep brain stimulation

University of Manitoba

Gender Dysphoria Assessment and Action for Youth: Review of Healthcare Services and Experiences of Trans Youth in Manitoba

Jack Heard

511

Ryan Henrie

512

University of Manitoba

Neuregulin-1 Promotes Myelin Repair Following Spinal Cord Injury

Nicholas Jette

513

University of Calgary

ATM deficiency and PARP inhibitors

Cameron Kaye

514

University of Manitoba

Two-stage reconstruction of complex dielectric permittivity and magnetic permeability for biomedical microwave imaging employing magnetic contrast agents

Katrice Kazmerik

515

University of Manitoba

Contrasting Drill Technique in Cadaveric and Printed Temporal Bone Simulation

Noren Khamis

516

University of British Columbia

Evaluating medical students’ interest and readiness for learning and practicing highvalue care: Preparing for change

Anna Kovalchuk

517

University of Calgary

Chemotherapy, tumours, and the brain.

Alexander Levit

518

Schulich School of Medicine & Dentistry

Novel analyses of cognitive flexibility in a transgenic hAPP rat model with comorbid striatal ischemic stroke

Donna Liao

519

University of British Columbia

Effectiveness of Interactive Medical Language Workshops for Helping Chinesespeaking Patients with Limited English Proficiency

Amber MacLellan

520

Dalhousie University Medical School

Fibre Intake Following Exclusive Enteral Nutrition (EEN) Therapy Alters Microbiome in Pediatric Crohn’s Disease Patients

Andrew McDermid

521

University of Manitoba

Genetic Diversity and Recombination in Circulating Serotypes from the Enterovirus B Species

University of Toronto

Slit2 Decreases the Size of Atherosclerotic Lesions in mice by Preventing Macrophage Oxldl Uptake and Foam Cell Formation

Ilya Mukovozov

522

Mathieu NadeauVallée

523

Université de Montréal

Antenatal suppression of interleukin-1 protects against inflammation-induced fetal injury and improves neonatal and developmental outcomes in mice

Alex Nantsios

524

University of Ottawa

Validation of Microfluidic Encapsulation of Cardiac Stem Cells for Myocardial Repair.

Siddharth Nath

525

McMaster University

Non-CAG triplet repeat expansion mutations in ataxin-7 and top1mt lead to ataxia

Vincent PicherMartel

526

Laval University

Mutation in Ubiquilin-2 exacerbates ALS/FTD features in a TDP-43 mouse model

Sheharyar Raza

527

University of Toronto

Life-Threatening Motor Vehicle Crashes Under Bright Sunlight

Quinlan Richert

528

University of Manitoba

Systemic inflammation before and after antiretroviral therapy initiation as a predictor of immune response among HIVinfected individuals in Manitoba

Nadine Rockwood

529

Memorial University of Newfoundland

Infertility and womenâ&#x20AC;&#x2122;s mental health: The relationship of female-factor infertility and depression and anxiety

Joanna Ryan

530

Schulich School of Medicine & Dentistry

The role of EZH2 in KRASG12D-mediated acinar-to-ductal metaplasia

Dalhousie University Medical School

Investigating ischemia-reperfusion (I/R) injury: Identification of a novel inflammatory axis induced by hypoxia in primary endothelial cells.

McGill University

Reflex testing for germline BRCA1, BRCA2, PALB2 and ATM mutations in pancreatic cancer

Nicholas Sajko

Alyssa Smith

531

532

Naveet Sohi

533

University of Alberta

MRI 3-Dimensional Texture Changes in the Motor Cortex Parallel Phenotypic Heterogeneity in Amyotrophic Lateral Sclerosis

Xiao Jing (Lily) Sun

534

University of Alberta

Total Arsenic and Speciation of Arsenic in Alberta Freshwater Fish

Daniyil Svystonyuk

535

University of Calgary

Acellular Matrix Biomaterial Promotes Cardiac Repair through an Active BioInductive Mechanism

Stephanie Totten

536

McGill University

The Yin-Yang of interferon gamma signaling in breast cancer immune surveillance and simultaneous metastatic progression.

Karim Wafa

537

Dalhousie University Medical School

Cellular physiology of iron in inflammation and infection processes: novel Fe chelator responses in experimental model systems

University of British Columbia

Common Pathogenic Mechanisms in Agerelated Macular Degeneration, Alzheimerâ&#x20AC;&#x2122;s Disease, Atherosclerosis and Glomerulonephritis

Alis (Qinyuan) Xu

538

Weining Yang

539

University of Toronto

Alterations in cervical glutamatergic interneurons preserves respiratory function in cervical spondylotic myelopathy (CSM)

Russell Yanofsky

540

McGill University

Sex differences in resistin, chemerin, and chemerin’s receptor in association with carotid atherosclerotic plaque instability

Albert Yeung

541

University of Manitoba

Contribution of Pannexin-1 activation to amyloid-beta induced synaptotoxicity and neurotoxicity

Université de Montréal

Discrepancy between cervical diagnostic biopsy and treatment histology: clinical and viral determinants

University of Toronto

Hyperconnectivity in stem cell-derived neurons of autistic children with SHANK2 haploinsufficiency.

University of Manitoba

Doxorubicin and Hypoxia Induced Mitochondrial Injury and Necrotic Cell Death of Ventricular Myocytes is Suppressed by Ellagic Acid

Nadège Zanré

Kirill Zaslavsky

Pegah Afshar

542

543

544

2:15 – 3:45 P.M. Session – Lightning Oral Presentations Procurity Lecture Theatre Main Floor – Apotex Centre Each presentation will be 6 minute for presentation and 4 minutes for questions. Chairs: TBA Student

Poster #

School

Abstract Title

Kirill Zaslavsky

543

University of Toronto

Hyperconnectivity in stem cell-derived neurons of autistic children with SHANK2 haploinsufficiency.

Vincent PickerMartel

526

Laval University

Mutation in Ubiquilin-2 exacerbates ALS/FTD features in a TDP-43 mouse model

University of Alberta

MRI 3-Dimensional Texture Changes in the Motor Cortex Parallel Phenotypic Heterogeneity in Amyotrophic Lateral Sclerosis

Naveet Sohi

533

Daniyil Svystonyuk

535

University of Calgary

Acellular Matrix Biomaterial Promotes Cardiac Repair through an Active BioInductive Mechanism

Stephanie Totten

536

McGill University

The Yin-Yang of interferon gamma signaling in breast cancer immune surveillance and simultaneous metastatic progression.

Ryan Henrie

512

University of Manitoba

Neuregulin-1 Promotes Myelin Repair Following Spinal Cord Injury

Alis (Qinyuan) Xu

538

University of British Columbia

Common Pathogenic Mechanisms in Agerelated Macular Degeneration, Alzheimer’s Disease, Atherosclerosis and Glomerulonephritis

Nadège Zanré

542

Université de Montréal

Discrepancy between cervical diagnostic biopsy and treatment histology: clinical and viral determinants

University of Toronto

Slit2 Decreases the Size of Atherosclerotic Lesions in mice by Preventing Macrophage Oxldl Uptake and Foam Cell Formation

Ilya Mukovozov

522

Lightning Oral Abstracts A.M. Session Procurity Lecture Theatre, Apotex Building Main Floor, Room 164 8:45 â&#x20AC;&#x201C; 9:55 a.m. Wednesday, June 7th, 2017

Abstract Title: Suicide risk correlates with lithium response in patients with bipolar disorder Authors: Nathan Corbett, Dr. Martin Alda Department: Dalhousie Psychiatry Abstract: Rationale: While lithium treatment is protective against suicide and suicide attempts (SA) in persons with mood disorders, it is not known whether the ability to respond to lithium, something that is thought to be biologically based, is predictive of SA risk. Objectives: To determine whether the quality of lithium response correlates with SA probability. Methods: Subjects participated in the NS and affiliated bipolar disorder (BPD) cohorts. Data for 102 individuals with a known SA were compared with 474 who did not by generalized linear modeling in R. Lithium response was quantified using a validated scale of clinical improvement, the Alda scale. Patients in the cohort were followed for a median of 22.79 years of observed illness. Results: Lithium response, as measured by the Alda scale, is correlated strongly with absence of SA (p=1.6e-04, z=-3.778), corrected for covariates. However, association of lithium response with total number of recorded SA was not significant (p=0.38). Age of first recorded SA did not significantly correlate with lithium response (p= 0.284). The covariates social anxiety, obsessive compulsive disorder, sleep disorder, family history of suicidality, generalized anxiety disorder (GAD), and panic disorder were all significantly correlated with SA. Conclusions: Risk of SA in long-term follow up of a bipolar disorders cohort correlates with observed response to lithium treatment, which responders having a lower chance of having one or more SA. Additionally, this study confirms prior association of anxiety disorders with SA in BPD. Funding: Faculty of Medicine Gladys Osman Summer Studentship 2015

Title: Orthotopic Patient Derived Xenografts as Relevant Models of Brain Metastasis Authors: Matthew Dankner1, Paul Savage1, April A.N. Rose1, Marie-Christine Guiot2, Morag Park1, Roberto J. Diaz3, Kevin Petrecca3, Peter M. Siegel1 1

Goodman Cancer Research Centre, 1160 Pine Avenue West, Montreal, Quebec, Canada, H3A 1A3. McGill University, MontrĂŠal, QuĂŠbec, Canada. 2

Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada; Division of Neuropathology, Department of Pathology, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada. 3

Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada Rationale: Brain metastasis occurs in 10-20% of cancer patients and results in median survival times of less than 1 year. In order to begin accurately developing novel treatment strategies, there is an urgent need to establish valid animal models of brain metastasis. Objectives: We sought to establish a biobank of orthotopic patient derived xenografts (PDX) models of brain metastasis from diverse solid cancers to be used to understand biological characteristics of brain metastasis and to test novel drug candidates in their ability to treat brain metastases. Methods: Surgical specimens of brain metastases from patients with diverse solid cancers were enzymatically dissociated and used to perform intra-cranial injections into NOD/scidgamma(c)null mice. At clinical endpoint, animals were sacrificed and brains were harvested for the following material: 1) formalin fixed paraffin embedded tissue, 2) viable frozen tissue, 3) snap frozen tissue and 4) material for dissociation and subsequent short-term tumorsphere culture and reimplantation. Cultured brain-metastatic cells were labeled with a ZsGreen/Luc dual reporter that permits longitudinal optical imaging following subsequent re-injection into mice. Results: 21 brain metastasis samples (7 lung primary, 8 breast, 4 melanoma, 1 colorectal, 1 gastric) have been implanted intra-cranially into mice. 13/21 (6 lung, 3 breast, 3 melanoma, 1 gastric) have successfully established tumours in the brains of mice as determined by hematoxylin and eosin (H&E) staining. Of these 13 metastases, 11 have successfully formed second-passage lesions in new recipient mice. The remaining 2 PDXs (2 lung primary) have not yet reached experimental endpoint. Analysis of H&E stained specimens derived from the surgical specimens and the PDX material reveals greater similarity between the patient sample and the orthotopic cranial PDX compared to subcutaneous (lung, melanoma, colorectal, gastric) or mammary fat pad (breast) xenografts. In order to apply these models to further develop biological understanding and treatment of brain metastasis, studies are underway to refine treatment for non-canonical BRAF mutant melanoma, as well as to characterize novel chemotherapeutic agents. Conclusions: PDXs of brain metastasis represent important models that can be employed to test novel therapeutics and to improve our understanding of the molecular mechanisms engaged by brain metastases. 21

Body mass index-for-age and weight-for-length < 2 years are associated with weight and cardiometabolic outcomes mid-childhood Kayla R. Furlong,a,b Laura N. Anderson,b,c David Dai,d Gerald LeBovic,d,e Patricia C. Parkin,b,e,f,g Jonathon L. Maguire,d,e,f,g,h,i Jeff Critch,a Catherine S. Birkenb,e,f,g, TARGet Kids! Collaboration AFFLIATIONS: a Department of Pediatrics, Memorial University, St. John’s, Newfoundland, Canada; b Child Health and Evaluative Sciences, Research Institute, and fPaediatric Outcomes Research Team, Division of Paediatric Medicine, Department of Paediatrics, The Hospital for Sick Children, Toronto, Ontario, Canada; c Department of Clinical Epidemiology and Biostatistics, Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada; e Institute of Health Policy, Management, and Evaluation, and Departments of g Paediatrics and hNutritional Sciences, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada; and dThe Applied Health Research Centre of the Li Ka Shing Knowledge Institute, and i Department of Pediatrics, St. Michael’s Hospital, Toronto, Ontario, Canada BACKGROUND: Weight status in children < 2 years has been associated with weight and cardiometabolic outcomes later in life; however, the most appropriate anthropometric tool to determine weight status in this age group is debated. It is recommended to plot weight-forlength for children <2 years of age and transition to BMI-for-age for children ≥ 2 years of age. Both tools perform similarly in classifying weight status for early screening, although it is unclear which measure is most appropriate to predict future outcomes. OBJECTIVES: The primary objective was to evaluate the association between BMI-for-age and, separately, weight-for-length in children < 2 years and BMI-for-age in mid-childhood (5 to < 9 years). The secondary objective was to evaluate these associations for cardiometabolic risk in mid-childhood. METHODS: We studied 2075 healthy, term-born children recruited from nine primary health practices in the TARGet Kids! research network in Toronto, Canada (July 2008 – July 2016). Length/height (cm) and weight (kg) measurements were obtained. For visits < 2 years, BMI-forage and weight-for-length was calculated. At mid-childhood, BMI-for-age was calculated. Blood samples were available for 483 children at mid-childhood. A continuous cardiometabolic risk score was calculated using age- and sex-standardized z-scores that included: waist circumference, high-density lipoprotein, blood glucose, systolic blood pressure, and triglycerides. Linear regression was used to examine the relation between exposure and outcome variables. R-squared values were calculated to estimate the proportion of variation explained by the models. RESULTS: BMI-for-age (r = 0.42, 95% CI: 0.36 – 0.44) and weight-for-length (r = 0.44, 95% CI: 0.40 – 0.48) at < 2 years were moderately associated with BMI-for-age at mid-childhood. Rsquared values were similar between BMI-for-age (R2 = 0.17) and weight-for-length (R2 = 0.20). BMI-for-age (r = 0.13, 95% CI: 0.04 – 0.21) and weight-for-length (r = 0.16, 95% CI: 0.07 – 0.24) at < 2 years were weakly associated with cardiometabolic risk at mid-childhood. R-squared values were similar between BMI-for-age (R2 = 0.02) and weight-for-length (R2 = 0.02). CONCLUSION: BMI-for-age and weight-for-length in children < 2 years are moderately associated with BMI-for-age and weakly associated with cardiometabolic risk in mid-childhood. Results suggest the use of either tool to estimate future weight and cardiometabolic outcomes. FUNDING: CIHR, SickKids Foundation, St. Michael’s Hospital Foundation.EMAIL: krf154@mun.ca 22

Robotic TMS mapping of motor cortex in the developing brain Grab JG, Zewdie E, Kuo HC, Giuffre A, Carlson H, and Kirton A University of Alberta Special Training in Research Program, Calgary Pediatric Stroke Program, Alberta Children’s Hospital; Alberta Children’s Hospital Research Institute (ACHRI), Calgary, AB; Introduction: Mapping motor cortex with transcranial magnetic stimulation (TMS) can inform brain plasticity and neurophysiology. TMS is safe and well tolerated in children but mapping is technically challenging and prone to human error. Emerging TMS robot technology has not been applied to motor mapping or children. We aimed to determine the feasibility and validity of robotic TMS motor mapping in children. Methods: Twelve typically developing, right-handed participants aged 9-18 years provided informed consent/assent. All underwent standardized MRI at 3T with T1 anatomical images uploaded to a neuronavigation system (Brainsight 2, Rogue). The participant, MRI, and TMS were co-registered within a TMS robotic system (Axilum). The motor hotspot and rest motor threshold for contralateral first dorsal interossei were mapped with surface EMG. An individualized grid (10X10, 7mm intervals) was centred over the hotspot. Responsive points were defined by having ≥2/4 suprathreshold single pulses produce an MEP amplitude >50µV. Outcomes included motor map graphs, area, volume, and centre of gravity. Bilateral motor function was determined using Perdue Pegboard Task (PPT). Safety and tolerability was evaluated. Results: Procedures were well tolerated with no adverse events. Two young subjects had high rest motor thresholds that precluded mapping. In the remaining 10, high resolution motor cortex maps were generated (see figure). Once thresholding was complete, motor mapping took 15-20 minutes per subject. Neuronavigation combined with near-real-time motion correction by the robotic arm provided high precision. Typical outcomes included motor map areas of 14.11±2.01 cm2 (range=8.33-29.89) and volumes of 0.81± 0.29 cm2V (range=0.28-3.34). No effects of age or gender were suggested. PPT score was possibly correlated with motor map volume (r=0.48, p= 0.16). Conclusion: Robotic TMS motor mapping is feasible, efficient, accurate and tolerated in children. The ability to quickly generate detailed, individualized maps suggests utility as a neurophysiological biomarker of motor cortex dysfunction and plasticity. Funding: Alberta Innovates Health Solutions (AIHS) jgrab@ualberta.ca

Gender Dysphoria Assessment and Action for Youth: Review of Healthcare Services and Experiences of Trans Youth in Manitoba Jack Heard BSc, Amanda Morris MD FRCSC, Nicole Kirouac RN BN, Jennifer Ducharme PhD C. Psych, Simon Trepel MD FRCPC, Brandy Wicklow MD MSc FRCPC

OBJECTIVES: To describe the pediatric transgender population accessing healthcare through the Manitoba Gender Dysphoria Assessment and Action for Youth (GDAAY) program, and report youthâ&#x20AC;&#x2122;s experiences accessing healthcare in Manitoba. METHODS: Demographic, medical, surgical, and mental health information was extracted from the medical records of youth referred to the GDAAY program (n=174). A 77-item online survey was conducted with a subset of those youth (n=25) to identify common healthcare experiences and perceptions of trans youth in Manitoba. RESULTS: Chart review of 122 natal females and 52 natal males, ranging in age from 4.7 to 17.8 years (mean 13.9 years), found sixty-six patients (46.8%) with a preexisting or current mental health diagnosis, of which anxiety and depression were the most common (n=43, 30.5%). Qualitative self-reports revealed all patients had negative interactions with healthcare providers at some point, many having experienced lack of engagement with the medical system due to reported lack of knowledge by the provider on trans related health services. CONCLUSION: Transgender youth in Manitoba seeking GDAAY services have high rates of anxiety and depression. These youth face adversity in healthcare settings and are distressed over long wait times for mental health services. Recommendations to improve care include increasing general health care providersâ&#x20AC;&#x2122; education on gender affirmative care, providing gender sensitivity training for healthcare providers, gathering preferred names and pronouns during triage, increasing visibility of support for LGBT+ persons in clinics, increasing resource allocation to this field, and creating policies so all healthcare settings are safe places for trans youth.

Alterations in cervical glutamatergic interneurons preserves respiratory function in cervical spondylotic myelopathy (CSM) W Yang1, K Satkunendrarajah2, SK Karadimas2,3, M Khazaei2, MG Fehlings1,2,3 1

Faculty of Medicine, University of Toronto, Toronto, ON, Canada Genetics and Development, Krembil Neuroscience Centre, Toronto, ON, Canada 3 Department of Neurosurgery, University of Toronto, Toronto, ON, Canada 2

Rationale: In contrast to traumatic spinal cord injury (SCI), chronic compressive disorders such as cervical spondylotic myelopathy (CSM) lead to a relative preservation of respiratory function. Using a murine model of CSM, we sought to investigate which components of the spinal respiratory networks contributed to maintaining respiration following graduated spinal cord compression. Objectives: To describe the spinal respiratory network alterations in CSM which allow the preservation of respiratory function Methods: To model CSM, mice were implanted with an aromatic polyether compression material underneath the C5-6 laminae. Sham animals were subject to C2 hemisection. Cholera toxin B (CTB) and PRV-152 were used, respectively, to label phrenic motoneurons (PMNs) and prephrenic interneurons. VGluT2-Cre//ROSA26 mice were used to identify excitatory glutamatergic spinal neurons. Diaphragmatic activity in both groups was monitored by electrophysiology. Spinal cord sections were then subject to immunofluorescence and unbiased stereological quantification. Results: Electrophysiological evaluation of C2 hemisection animals revealed complete paralysis of the ipsilateral hemidiaphragm, whereas CSM animals had preserved respiratory function at 8 weeks. Both groups showed a loss of Cholera Toxin B-labeled phrenic motoneurons (PMNs). Notably, CSM animals showed an increase in PRV-152-positive cervical prephrenic interneurons, as well as an increase in dendritic arborization of these interneurons. Using VGluT2-Cre//ROSA26 mice, we further found that most of these PRV-152-positive prephrenic interneurons were double labeled with ROSA26-tdTomato and could thus be identified as glutamatergic in nature. Conclusion: Relative to C2 hemisection, CSM induction in mice was followed by compensatory prephrenic glutamatergic input onto PMNs, which contributed to a relative preservation of respiratory function. Understanding the alterations in spinal respiratory networks that preserve respiration in chronic compressive disorders will guide treatment for SCI patients facing respiratory insufficiency.

Funding: PVA, NREF, CREMS Research Scholar Program

Email: weining.yang@mail.utoronto.ca

Chemotherapy, tumours, and the brain. Anna Kovalchuk1, Yaroslav Ilnytskyy2, Rocio Rodriguez-Juarez2, Svitlana Shpyleva3, Stepan Melnyk3, Amanda Katz4, David Sidransky4, Olga Kovalchuk2 and Bryan Kolb1 1

Department of Neuroscience, 2Department of Biological Sciences, University of Lethbridge. Lethbridge, Canada. 3 Division of Biochemical Toxicology, National Center for Toxicological Research, FDA. Jefferson, USA. 4Department of Oncology, Champions Oncology. Baltimore, USA

Rationale: Many central nervous system complications arise and persist because of chemotherapy treatment. This condition is known as chemo brain and affects over half of cancer survivors. Based on literature and our preliminary data, we propose a new epigenetic theory of chemo brain, whereby the mechanisms underlying chemotherapyâ&#x20AC;&#x2122;s neurotoxic side effects are epigenetic and associated with aberrant global gene expression. Objectives: To determine the effects of chemotherapy drugs on the brains of chemotherapytreated tumour-bearing mice, as compared to their untreated tumour-bearing and intact control counterparts. Methods: We looked at the prefrontal cortex (PFC) because of its role in various executive functions and published data showing it is sensitive to chemotherapy. We used mouse TumorGraft TM models of triple negative (TRN) and progesterone receptor positive (PR+) breast cancer. We profiled the epigenetic, transcriptome, and small RNAome changes in chemotherapy-treated tumor-bearing mice, as compared to untreated tumor-bearing mice and to intact control mice. Results: Tumor presence alone affected molecular networks and caused oxidative stress, manifesting as elevated levels of 8-oxodG. This was paralleled by global DNA methylation loss and an increase in DNA hydroxymethylation. Tumor growth also led to transcriptome changes, affecting several pathways. Chemotherapy treatment further intensified tumor-induced changes. Both tumor presence and chemotherapy treatment affected the small RNAome, of which miRNA changes were the most pronounced. Several miRNA families were deregulated in both tumorbearing and chemotherapy-treated animals and miRNA deregulation was associated with altered BDNF (brain-derived neurotrophic factor) levels and other important neural proteins. Conclusions: In sum, our results show that the presence of the tumor itself profoundly influenced molecular processes and networks in the brain, and chemotherapy exposure further exacerbated these changes. Upon completion, this project will delineate the molecular and cellular mechanisms of tumor brain and chemo brain that may underlie neuroanatomical changes and behavioral outcomes, and define chemo brain and tumor brain biomarkers. The outcomes of our study may be used to design strategies to mitigate tumor brain and chemo brain side effects. Funding: Supported by the CIHR grant. A.K. is a recipient of the Canada Vanier Graduate Scholarship and the AI-HS Graduate Scholarship. Email: anna.kovalchuk@uleth.ca

Poster Discussion Brodie Centre Mezzanine 10:00 â&#x20AC;&#x201C; 11:45 A.M.

INSTITUTIONAL PRACTICES AND PREDICTORS OF SURVIVAL FOR LIMITED-STAGE SMALL CELL LUNG CANCER â&#x20AC;&#x201C; A RETROSPECTIVE ANALYSIS Emma Ali1, BSc, Hanbo Chen2, MD, Andrew Warner2, MSc, George B. Rodrigues2, MD, Alexander V. Louie2, MD 1

Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada 2 Department of Radiation Oncology, London Regional Cancer Program, London Health Sciences Centre, London, Ontario, Canada Rationale: Three of ten patients diagnosed with small cell lung cancer (SCLC) present with limited-stage disease (LS-SCLC). Mortality rates are high due to aggressive recurrence and increased risk of metastases. A retrospective consecutive database for LS-SCLC patients was constructed at a Canadian tertiary care institution, and herein we report our institutional practices and outcomes. Objectives: We hypothesize that baseline patient and disease characteristics are inherently prognostic and predict treatment outcomes. Methods: Following Institutional Review Board approval, SCLC patient charts between January 1, 2000 and December 31, 2013 were retrospectively reviewed. Demographics, baseline performance status, disease presentation, therapy, toxicities, and outcome-related data were extracted for LS-SCLC patients and entered into an Access database (Microsoft, Redmond, WA). Summary statistics, exploratory univariate Cox proportional hazard and univariate logistic regression modeling were performed using SAS software (v9.4) with two-sided statistical testing at the 0.05 significance level. Results: Of the 1,131 charts reviewed, 398 patients (35%) were diagnosed with LS-SCLC. Median follow-up was 8.2 years and mean age at registration was 67 (range 40-87). Chemotherapy was utilized for 94.2% of patients, and radiotherapy for 81.7%. Patients received 4.9 + 1.63 (mean + SD) cycles of first-line chemotherapy and 47.2 + 12.6 Gy of radiotherapy. Of patients receiving both treatments, 81.2% had concurrent chemoradiation. Nineteen (4.8%) patients were initially treated with surgery. Prophylactic cranial irradiation (PCI) was provided to 45.2% of patients. The cohortâ&#x20AC;&#x2122;s 5-year overall survival (OS) and progression-free survival (PFS) was 19.7% and 14.1%, respectively. Median OS and PFS were 15.4 and 9.3 months, respectively. On univariate analysis, baseline characteristics such as age, performance status, forced expiratory volume, carbon monoxide diffusing capacity, T-stage, M-stage, and presence of pleural effusions were significantly associated with OS. Conclusions: Our institutional practice for this LS-SCLC cohort of chemotherapy with concurrent, conventionally-fractionated radiotherapy followed by PCI resulted in a 5-year OS comparable to previously-reported results. Multiple patient and disease-related factors were associated with survival on univariate analysis. Further analysis of relationships within the data is presently underway with the goal of creating a LS-SCLC-specific prognostic system.

To examine the mechanisms through which adipose-derived stem cells affect healthy and malignant breast epithelial cell proliferation Berdnikov A1, Chatterjee S2, Buchel E1, Raouf A2, Lee-Wing V2, Safneck J3 1) University of Manitoba Department of Surgery, Section of Plastic Surgery 2) Research Institute in Oncology and Hematology, and 3) Department of Pathology Winnipeg, MB, Canada; University of Manitoba, Winnipeg, MB, Canada

Introduction: Mastectomies are commonly performed procedures to increase the life expectancy of women suffering from breast cancer. Stromal Vascular Fraction (SVF) from patient’s abdominal fat is often utilized to supplement fat grafts used in breast reconstructions. Unfortunately, very little is known about the interaction of SVF with residual cancer cells as well as surrounding breast tissue. This study investigates if secreted factors from the SVF cells affect proliferation of residual breast cancer cells and the surrounding breast cells. Methods: The effects of patient-derived SVF samples on the proliferation of estrogen receptor positive (ER+) MCF-7 breast cancer cells, tissue adjacent to ER+ breast tumours (TAT), and healthy breast cells from reduction mammoplasty samples (HBT) was examined. The secretion profile of 41 different cytokines in the conditioned media (CM) of SVF samples grown in cocultures with the MCF-7, TAT, or HBT cells was compared to the CM of each cell type grown alone. One-way analysis of variance with Tukey’s multiple comparisons test were performed for multiple pair-wise comparisons. Single pair-wise comparisons were performed using the two-tail student t-test. Results: Three TAT, SVF and HBT samples were surgically obtained from three separate consented patients for use during this experiment. In addition, Michigan Cancer Foundation-7 (MCF-7) cells were used to mimic invasive ductal cell carcinoma cells. Placing MCF-7 cells in co-cultures with SVF cells led to a 1.53-fold (p<0.005) increase in their proliferation. Interestingly, CM obtained from the co-cultures of SVF+MCF-7 cells was sufficient to increase MCF-7 cell proliferation by 1.5 fold (p<0.05), indicating that secreted factors in this CM have pro-proliferative properties on cancer cells. The cytokine array identified increased secretion of a number of cytokines that were uniquely elevated in the co-cultures of SVF with MCF, TAT or HBT cells but not in the individual cell cultures. From the cytokine array data, we confirmed that IL-1β, MDC and RANTES cytokines were able to increase MCF-7 cell proliferation independently. Conclusion: Secreted factors from SVF+MCF-7 co-cultures have pro-proliferative effects on MCF-7 cells. This study is a proof of concept that examining secretion profiles of SVF cocultures can lead to the discovery of cytokines which are able to modulate breast cancer cell proliferation.

Application and Usefulness of Outpatient Cardiac Testing among Emergency Department Patients with Syncope Olivia G. Cook,2, Muhammad A. Mukarram2, Soo-Min Kim,2, Kirtana Arcot,2, Monica Taljaard2,3, Marco L.A. Sivilotti4,5, Brian H. Rowe6, Venkatesh Thiruganasambandamoorthy2,3,7 1

Faculty of Medicine, University of Ottawa, Ottawa, Canada Ottawa Hospital Research Institute, Ottawa, Ontario, Canada 3 School of Epidemiology, Public Health and Preventive Medicine, University of Ottawa, Ottawa, Ontario, Canada 4 Department of Emergency Medicine, Queenâ&#x20AC;&#x2122;s University, Kingston, Ontario, Canada 5 Department of Biomedical and Molecular Sciences, Queenâ&#x20AC;&#x2122;s University, Kingston, Ontario, Canada 6 Department of Emergency Medicine and School of Public Health, University of Alberta, Edmonton, Alberta, Canada 7 Department of Emergency Medicine, University of Ottawa, Ottawa, Ontario, Canada 2

Rationale: 2.6% of emergency department (ED) syncope patients will have underlying cardiac serious conditions (e.g. arrhythmia, serious structural heart disease) identified within 30-days of disposition. If those at risk are discharged home, outpatient cardiac testing can detect underlying arrhythmias and structural heart disease, and thereby improve patient safety. Objectives: We describe the frequency of outpatient referrals for cardiac testing and the proportion of cardiac serious adverse events (SAE) among high risk and non-high (low and medium) risk ED syncope patients, as defined by the Canadian Syncope Risk Score (CSRS). Methods: We conducted a multicenter prospective cohort study to enroll adult syncope patients across five large tertiary care EDs. We collected demographics, medical history, disposition, CSRS value, outpatient referrals and testing results (holter, echocardiography), and cardiac SAE. Adjudicated 30-day SAE included death due to unknown cause, myocardial infarction, arrhythmia, and structural heart disease. We used descriptive analysis. Results: Of 4064 enrolled patients, a total of 955 patients (23%) received an outpatient referral (mean age 57.7 years, 52.1% female). Of the 299 patients (7%) hospitalized, 154 received outpatient cardiac testing after discharge. Among the 3765 patients discharged home from the ED, 40% of the non-high risk patients (305/756) and 56% of the high-risk patients (25/45) received outpatient cardiac testing. Of all patients who received outpatient cardiac testing, 4 patients (0.8%) had serious cardiac conditions identified and all were arrhythmias. Among those with no cardiac testing, 5 patients (0.9%) suffered cardiac SAE (80% arrhythmias) outside the hospital. Of the 20 (44%) high risk patients who did not receive outpatient cardiac testing, 2 (10%) patients suffered arrhythmias outside the hospital. While among the 451 non-high risk patients, only 0.8% suffered arrhythmia outside the hospital. Conclusion: Outpatient cardiac testing among ED syncope patients is largely underutilized, especially among high risk patients. Better guidelines for outpatient cardiac testing are needed, as current practice is highly variable and mismatched with patient risk. Email: ocook088@uottawa.ca

Abstract Title: Suicide risk correlates with lithium response in patients with bipolar disorder Authors: Nathan Corbett, Dr. Martin Alda Department: Dalhousie Psychiatry Rationale: While lithium treatment is protective against suicide and suicide attempts (SA) in persons with mood disorders, it is not known whether the ability to respond to lithium, something that is thought to be biologically based, is predictive of SA risk. Objectives: To determine whether the quality of lithium response correlates with SA probability. Methods: Subjects participated in the NS and affiliated bipolar disorder (BPD) cohorts. Data for 102 individuals with a known SA were compared with 474 who did not by generalized linear modeling in R. Lithium response was quantified using a validated scale of clinical improvement, the Alda scale. Patients in the cohort were followed for a median of 22.79 years of observed illness. Results: Lithium response, as measured by the Alda scale, is correlated strongly with absence of SA (p=1.6e-04, z=-3.778), corrected for covariates. However, association of lithium response with total number of recorded SA was not significant (p=0.38). Age of first recorded SA did not significantly correlate with lithium response (p= 0.284). The covariates social anxiety, obsessive compulsive disorder, sleep disorder, family history of suicidality, generalized anxiety disorder (GAD), and panic disorder were all significantly correlated with SA. Conclusions: Risk of SA in long-term follow up of a bipolar disorders cohort correlates with observed response to lithium treatment, which responders having a lower chance of having one or more SA. Additionally, this study confirms prior association of anxiety disorders with SA in BPD. Funding: Faculty of Medicine Gladys Osman Summer Studentship 2015

Goodman Cancer Research Centre, 1160 Pine Avenue West, Montreal, Quebec, Canada, H3A 1A3. McGill University, MontrĂŠal, QuĂŠbec, Canada. 2 Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada; Division of Neuropathology, Department of Pathology, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada. 3 Department of Neurology and Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, 3801 University St., Montreal, QC, H3A 2B4, Canada

Rationale: Brain metastasis occurs in 10-20% of cancer patients and results in median survival times of less than 1 year. In order to begin accurately developing novel treatment strategies, there is an urgent need to establish valid animal models of brain metastasis. Objectives: We sought to establish a biobank of orthotopic patient derived xenografts (PDX) models of brain metastasis from diverse solid cancers to be used to understand biological characteristics of brain metastasis and to test novel drug candidates in their ability to treat brain metastases. Methods: Surgical specimens of brain metastases from patients with diverse solid cancers were enzymatically dissociated and used to perform intra-cranial injections into NOD/scidgamma(c)null mice. At clinical endpoint, animals were sacrificed and brains were harvested for the following material: 1) formalin fixed paraffin embedded tissue, 2) viable frozen tissue, 3) snap frozen tissue and 4) material for dissociation and subsequent short-term tumorsphere culture and reimplantation. Cultured brain-metastatic cells were labeled with a ZsGreen/Luc dual reporter that permits longitudinal optical imaging following subsequent re-injection into mice. Results: 21 brain metastasis samples (7 lung primary, 8 breast, 4 melanoma, 1 colorectal, 1 gastric) have been implanted intra-cranially into mice. 13/21 (6 lung, 3 breast, 3 melanoma, 1 gastric) have successfully established tumours in the brains of mice as determined by hematoxylin and eosin (H&E) staining. Of these 13 metastases, 11 have successfully formed second-passage lesions in new recipient mice. The remaining 2 PDXs (2 lung primary) have not yet reached experimental endpoint. Analysis of H&E stained specimens derived from the surgical specimens and the PDX material reveals greater similarity between the patient sample and the orthotopic cranial PDX compared to subcutaneous (lung, melanoma, colorectal, gastric) or mammary fat pad (breast) xenografts. In order to apply these models to further develop biological understanding and treatment of brain metastasis, studies are underway to refine treatment for non-canonical BRAF mutant melanoma, as well as to characterize novel chemotherapeutic agents. Conclusions: PDXs of brain metastasis represent important models that can be employed to test novel therapeutics and to improve our understanding of the molecular mechanisms engaged by brain metastases.

Espace Transition au Musée – Deconstructing stigma one painting at time Victor De Broux-Leduc1, Kim Archambault2, Patricia Garel 2,3, Sylvie Gauthier2, Elyse PorterVignola3

Medicine Department – University of Montreal, 2Psychiatric Department – Ste-Justine University Hospital, Montreal, 3Psychology Department – University of Quebec in Montreal 1

Background: The psychosocial rehabilitation of youths with mental disorders may be partly achieved through fighting stigmatisation and improving self-perception. This poster reports on the preliminary evaluation of Espace Transition au Musée1 (ETM), an innovative art-based group program that aims to foster the well-being and the psychosocial adjustment of youths with mental disorders. Aims: Objectives were to assess the program effects related to self-perception and stigma. Methods: ETM targeted participants aged 14 to 25 years. Group included youths presenting with psychopathology (2/3) and same-aged peers with no history of mental disorder (1/3). The program took place at the Montreal Museum of Fine Arts once a week during twelve weeks. Sessions were conducted by a non-therapist art educator and included art interpretation and collaborative artmaking workshops. Clinical support was provided on-site by a mental health practitioner. Mixed methods evaluation data were collected before, during and up to three months following program completion from 16 participants, their close relatives (N=12), referring clinicians (N=12), and program staff (N=2) using standardized questionnaires, semi-structured qualitative interviews, and direct observations. Results: Preliminary quantitative results suggest significant reduction in negative self-view and perceived stigma following program participation (all p<0.05). Conclusions: This evaluation suggests that ETM represents a promising strategy to support rehabilitation of youths with mental disorders through improvement of self-perception and reduction of stigmatisation. Further analysis, including qualitative, will help identifying the specific program components and mechanisms that may be responsible for such benefits. Funding: CHU Sainte-Justine Foundation

E-mail: victor.de.broux-leduc@umontreal.ca

Original French name; could be translated as: Transition Space at the Museum 33

of Pediatrics, Memorial University, St. John’s, Newfoundland, Canada; Health and Evaluative Sciences, Research Institute, and fPaediatric Outcomes Research Team, Division of Paediatric Medicine, Department of Paediatrics, The Hospital for Sick Children, Toronto, Ontario, Canada; cDepartment of Clinical Epidemiology and Biostatistics, Faculty of Health Sciences, McMaster University, Hamilton, Ontario, Canada; eInstitute of Health Policy, Management, and Evaluation, and Departments of gPaediatrics and hNutritional Sciences, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada; and dThe Applied Health Research Centre of the Li Ka Shing Knowledge Institute, and iDepartment of Pediatrics, St. Michael’s Hospital, Toronto, Ontario, Canada bChild

BACKGROUND: Weight status in children < 2 years has been associated with weight and cardiometabolic outcomes later in life; however, the most appropriate anthropometric tool to determine weight status in this age group is debated. It is recommended to plot weight-forlength for children <2 years of age and transition to BMI-for-age for children ≥ 2 years of age. Both tools perform similarly in classifying weight status for early screening, although it is unclear which measure is most appropriate to predict future outcomes. OBJECTIVES: The primary objective was to evaluate the association between BMI-for-age and, separately, weight-for-length in children < 2 years and BMI-for-age in mid-childhood (5 to < 9 years). The secondary objective was to evaluate these associations for cardiometabolic risk in mid-childhood. METHODS: We studied 2075 healthy, term-born children recruited from nine primary health practices in the TARGet Kids! research network in Toronto, Canada (July 2008 – July 2016). Length/height (cm) and weight (kg) measurements were obtained. For visits < 2 years, BMI-forage and weight-for-length was calculated. At mid-childhood, BMI-for-age was calculated. Blood samples were available for 483 children at mid-childhood. A continuous cardiometabolic risk score was calculated using age- and sex-standardized z-scores that included: waist circumference, high-density lipoprotein, blood glucose, systolic blood pressure, and triglycerides. Linear regression was used to examine the relation between exposure and outcome variables. R-squared values were calculated to estimate the proportion of variation explained by the models. RESULTS: BMI-for-age (r = 0.42, 95% CI: 0.36 – 0.44) and weight-for-length (r = 0.44, 95% CI: 0.40 – 0.48) at < 2 years were moderately associated with BMI-for-age at mid-childhood. Rsquared values were similar between BMI-for-age (R2 = 0.17) and weight-for-length (R2 = 0.20). BMI-for-age (r = 0.13, 95% CI: 0.04 – 0.21) and weight-for-length (r = 0.16, 95% CI: 0.07 – 0.24) at < 2 years were weakly associated with cardiometabolic risk at mid-childhood. R-squared values were similar between BMI-for-age (R2 = 0.02) and weight-for-length (R2 = 0.02). CONCLUSION: BMI-for-age and weight-for-length in children < 2 years are moderately associated with BMI-for-age and weakly associated with cardiometabolic risk in mid-childhood. Results suggest the use of either tool to estimate future weight and cardiometabolic outcomes. FUNDING: CIHR, SickKids Foundation, St. Michael’s Hospital Foundation EMAIL: krf154@mun.ca 34

Survival of ventral midbrain dopaminergic neurons during aging is not regulated by netrin receptor UNC5A Louis Gervais1, Timothy E Kennedy2, Abbas F Sadikot2. 1

Groupe de recherche sur le système nerveux central, Université de Montréal. Montreal, Canada. 2

Department of Neurology and Neurosurgery, McGill University. Montreal, Canada.

Rationale: Netrin-1 and its receptor DCC are essential for the development and organization of dopaminergic nuclei in the ventral midbrain. The netrin receptor Unc5A regulates cell and axon guidance cue during neural development, and has also been hypothesized to act as a dependence receptor, regulating neuronal survival. Polymorphisms in the human Unc5A gene are associated with changes in the onset and severity of Parkinson’s disease, suggesting a possible role in the development and survival of dopaminergic neurons. Objectives: Determine the contribution of Unc5A to the survival of ventral midbrain dopaminergic neurons during development and aging. Methods: Using unbiased stereology, the number of tyrosine hydroxylase-positive dopaminergic cells in the ventral midbrain nuclei was counted in Unc5A null, heterozygous and wild-type mice, at 3 months and 12 months of age. Behaviour was evaluated using open-field and rotarod tests. Dopaminergic neuronal morphology and distribution was also evaluated. Results: No significant differences were detected in the morphology, distribution or number of dopaminergic cells in the ventral midbrain of Unc5A knockout mice at 3 and 12 months of age. Behavioural studies revealed no differences between genotypes in performance on the rotarod, while heterozygous mice exhibited modest hypoactivity in open field experiments. Conclusions: Unc5A is not essential for the development or survival of ventral midbrain dopaminergic neurons. Funding: CIHR, Parkinson Society Canada

Association between physical activity and cardiac risk factors in disease prevention within cardiac rehabilitation populations. Andrew Hannaa, Dr. Chris Blanchardb Dalhousie Medical Schoola, Dalhousie University. Halifax, Nova Scotia, Canada. Department of Medicineb, Dalhousie University. Halifax, Nova Scotia, Canada. Rationale: Although physical activity (PA) is a known factor in preventing secondary cardiovascular disease (CVD)1, the impact of PA (as defined by steps/day) on CVD risk factors is uncertain. Additionally, Ayabe et al. have proposed step count recommendations specifically for the secondary prevention and regression of CVD within cardiac rehabilitation (CR) patients2. However, no studies have assessed these step count recommendations and their association with CVD risk factors in the CR population. Objectives: The purposes of this study are to: a) examine the association between Ayabe et al. step count recommendations and CVD risk factors2, and b) to assess the continuous relationship between steps/day and CVD risk factors, all in the CR population. Methods: A total of 216 subjects participated in the study. Demographic and clinical data were collected through chart review. Physical activity was defined as steps/day and was measured by a pedometer. The CVD risk factors explored were waist circumference, body mass index, blood pressure, glucose, lipid profile, anxiety, and depression. Results: After adjustment for confounders, statistical significance was seen between the steps/day categories and: waist circumference (p=0.014), BMI (p=0.023), fasting glucose (p=0.017), triglycerides (p=0.024), HDL-C (p=0.026), and cholesterol/HDL-C ratio (p=0.005). Furthermore, a linear association was seen between continuous steps/day and waist circumference, glucose, cholesterol, triglycerides, HDL-C, cholesterol/HDL ratio, and depression. Conclusions: Both a step count category approach and a continuous steps/day approach showed evidence of reducing CVD risk factors. Physicians and CR programs can recommend either approach as appropriate. Funding: Department of Medicine University Internal Medicine Research Foundation (UIMRF) Studentship; Dalhousie Medicine Travel Fund. Email: andrew.hanna@dal.ca References: 1. Taylor, Rod S., et al. "Mortality reductions in patients receiving exercise-based cardiac

rehabilitation: how much can be attributed to cardiovascular risk factor improvements?." European Journal of Cardiovascular Prevention & Rehabilitation 13.3 (2006): 369-374. 2. Ayabe, Makoto, et al. "Target step count for the secondary prevention of cardiovascular disease." Circulation Journal 72.2 (2008): 299-303.

Use of magnetoencephalography to examine cortical responses to deep brain stimulation. Irene E. Harmsena, Nathan C. Rowlandb, Luis G. Dominguezc and Andres M. Lozanoa a

Division of Neurosurgery, Department of Surgery, University of Toronto, Krembil Neuroscience Centre, University Health Network, Toronto Western Hospital, Toronto, Ontario, Canada. b Department of Neurosurgery, Medical University of South Carolina, Charleston, South Carolina, USA. cDivision of Neurology, Department of Medicine, University of Toronto, Krembil Neuroscience Centre, University Health Network, Toronto Western Hospital, Toronto, Ontario, Canada. Rationale: Deep brain stimulation (DBS) is being applied to motor circuit disorders such as Parkinson’s disease and dystonia, to mood circuits to treat depression, and to cognitive circuits to treat Alzheimer’s disease. The mechanism by which DBS works is unknown, resulting in DBS programming to remain an empirical exercise requiring frequent patient visits that lead to suboptimal therapeutic outcomes. Since brain regions that are activated by the focal delivery of electrical stimulation in the brain are not well understood, this project examines the relationship between implant location, stimulation dosage, current frequency and brain activation. Objectives: We assessed the feasibility of collecting neurophysiological data in patients with active DBS systems during both resting state and task-based conditions. To optimize DBS programming we determined the relationship between cortical physiology, DBS parameter changes and clinical responses. Methods: Magnetoencephalography (MEG) is a non-invasive technique for mapping brain activity by recording magnetic fields produced by electrical currents in the brain. MEG was used in DBS patients to obtain both spatial and temporal measures of neuronal activity in response to various stimuli. Results: DBS patients have been recorded from a wide spectrum of neurological and psychiatric disorders including Parkinson’s disease, essential tremor, treatment-resistant depression, obsessive-compulsive disorder, anorexia, chronic pain, and Alzheimer’s disease. The data provides evidence that MEG in DBS patients is safe, feasible and is able to demonstrate specific activation of brain circuits. Conclusions: This is an extremely novel project since very few centres in the world are performing DBS surgery in such a diverse set of patients involving a wide variety of disorders and brain targets. This project provides valuable insight into the effects of DBS by comparing brain activation across various pathological states and determining the consequences of stimulating different neuronal circuits. These research findings help to optimize individual DBS settings based on patients’ neural activity and their clinical response, resulting in fewer clinical visits to adjust DBS parameters and an improvement in patient quality of life. Funding: CIHR. Email: irene.harmsen@mail.utoronto.ca 38

Neuregulin-1 Promotes Myelin Repair Following Spinal Cord Injury. Ryan Henrie, Arsalan Alizadeh, Hardeep Kataria, Kallivalappil T. Santhosh and Soheila KarimiAbdolrezaee Introduction: Spinal Cord Injury (SCI) is a devastating event which causes lifelong disability and results in significant emotional and financial burden for patients, their families, and the healthcare system. There are currently no therapies which can effectively improve the outcomes of SCI in patients. At the tissue level, SCI causes loss of oligodendrocytes, demyelination and concomitant axon degeneration. Moreover, activation of astrocytes leads to the formation of glial scar. Our lab has shown for the first time that a growth factor named neuregulin-1 (Nrg-1) is severely downregulated following SCI. These pathologic changes are progressive and permanent, and cause neurological disability with little or no functional recovery. Methods: A well-established and clinically relevant model of incomplete compressive SCI at mid-thoracic level was induced in adult female Sprague Dawley rats. Rats were assigned to receive either vehicle or Nrg-1 administered to the subarachnoid space using indwelling catheter. 10 weeks post-injury, spinal cords were sectioned and analyzed for confocal immunohistochemistry and semi-thin analysis of myelin indexing. Oligodendrocyte precursor cells isolated from mixed glial cultures from postnatal rat cortex (1-3 days) were used for in vitro experiments. T-tests and two-way ANOVA were used for statistical comparisons where appropriate. Results: Our Nrg-1 therapy significantly restored of the deficient levels of Nrg-1 in the injured spinal cord. Nrg-1 treatment improved myelin thickness around the injured axons at the injury epicenter and preserved mature oligodendrocytes in the area surrounding the injury site, and caused maturation of oligodendrocytes in vitro. Furthermore, Nrg-1 reduced axon degeneration, preserved neurons and reduced glial scar in the spinal cord lesion. Conclusion: For the first time, we have demonstrated positive modulatory effects of Nrg-1 on the post-SCI microenvironment at the chronic stage of injury. These results suggest the potential of Nrg-1 therapy to promote tissue repair following SCI, and its promise as a part of combinatorial approaches in regenerative medicine strategies including stem cell therapies. Funding: CIHR, University of Manitoba, Canadian Paraplegic Association, American Academy of Neurology, Childrenâ&#x20AC;&#x2122;s Hospital Research Foundation of Manitoba. Email: Soheila.Karimi@umanitoba.ca

Understanding PARP inhibitor sensitivity in ATM deficient cancer cells Nicholas Jette1,2,5, Chen Wang1,2,5, Arjumand Wani1,2,5, Danylo Moussienko1,2,5, Siddartha Goutam1,2,5, Elias Saba1,2, Paul Gordon3, Gwynn Bebb1,4,5, and Susan Lees-Miller1,2,5. (1) Robson DNA Science Center, University of Calgary (2) Department of Biochemistry and Molecular Biology, University of Calgary (3) Alberta Childrenâ&#x20AC;&#x2122;s Hospital Research Institute, University of Calgary (4) Department of Oncology, Tom Baker Cancer Center, Calgary, AB (5) Arnie Charbonneau Cancer Institute, University of Calgary. Background: Ataxia Telangiectasia Mutated (ATM) is an apex signaling kinase that plays a major role in the cellular response to DNA double strand breaks (DSBs). The Lees-Miller lab has previously shown that catalytic inhibition of ATM by small molecule inhibitor or depletion of ATM by shRNA, sensitizes cancer cells to the Poly ADP-Ribose Polymerase (PARP) inhibitor olaparib in vitro and in vivo in animal models. While it is clear that ATM deficient cells are sensitive to PARP inhibitors, the reason for sensitivity remains elusive. Results: Colorectal cancer cells with depletion of ATM are sensitive to olaparib. shATM HCT116 colorectal cancer cells accumulate in G2 following multiple rounds of replication but do not apoptose when treated with olaparib for up to 120 hours. Olaparib acts as a temporary antiproliferative agent in shATM HCT116 cells while WT HCT116 cells are unaffected by olaparib. Conclusion: Understanding how ATM deficient cancer cells respond to PARP inhibitor treatment has the potential to influence the more than 100 current clinical trials involving PARP inhibitors. This research could influence future PARP inhibitor regimens given to cancer patients and improve outcomes.

Two-stage reconstruction of complex dielectric permittivity and magnetic permeability for biomedical microwave imaging employing magnetic contrast agents Cameron Kayea, Ian Jeffreya and Joe LoVetria a Department of Electrical and Computer Engineering, University of Manitoba, Winnipeg, MB, Canada Background: Microwave imaging (MWI) has seen steady advancement towards specialized biomedical applications for several years, particularly in the field of diagnostic breast imaging. In addition to imaging tissuesâ&#x20AC;&#x2122; gigahertz-range complex permittivity profiles using conventional MWI techniques, recent studies have shown that magnetic contrast media such as magnetic nanoparticles (MNP) can augment the complex magnetic permeability of bodily tissues in which they accumulate, and that this quantitative property can also be successfully imaged using appropriately modified contrast-enhanced MWI algorithms. The combination of both properties would theoretically enrich the diagnostic utility of MWI as a future clinical modality. Methods: An implementation of the Contrast Source Inversion algorithm employing the discontinuous Galerkin method (DGM-CSI) has been modified to produce quantitative 2D and 3D images of both the dielectric permittivity and magnetic permeability of contrast-enhanced synthetic breast models and experimental targets. A reconstruction of the targetâ&#x20AC;&#x2122;s permittivity profile is obtained first and used as an inhomogeneous background for a second-stage reconstruction of the magnetic permeability only, based on differential data obtained through the use of a static magnetic field, or a simulation thereof. Numerical validation of the algorithm is carried out using finite-element breast models containing MNP-embedded tumours with simulated magnetic properties. Simple 2D polyethylene targets containing suspensions of magnetite MNPs at concentrations from 10 to 50 mg/mL were used for experimental validation as well, using data collected with a dipole antenna array. More realistic breast phantoms were also imaged in a newer 3D faceted resonant imaging chamber employing arrays of low-profile magnetic field probes. Results: Previous studies have shown that reconstructions of 2D synthetic data from models containing inclusions of MNPs at concentrations as low as 10 mg/mL can reliably detect and localize masses as small as 1.0 cm in diameter. Inversions of experimental data collected from crude 2D polyethylene targets containing suspensions of magnetite MNPs have allowed detection of inclusions as small as 1.27 cm in diameter, but their correct localization has been inconsistent, likely due to the poor sensitivity of the dipole antennas. Simulations and experiments involving full 3D breast models and phantoms are ongoing. Conclusions: Simulated and experimental imaging of dielectric permittivity and magnetic permeability continue to demonstrate the plausibility of incorporating MNPs in MWI and are helping to delineate a future role for microwave imaging in clinical breast health assessment.

Funding: Vanier CIHR, NSERC.

Contrasting Drill Technique in Cadaveric and Printed Temporal Bone Simulation Katrice Kazmerik1, Jordan Hochman2,5, Justyn Pisa,2,5, Lorenzo Gentile3, Bertram Unger4,5 1

Faculty of Health Sciences, 2Department of Otolaryngology, 3Faculty of Computer Science, Department of Internal Medicine, University of Manitoba, 5Laboratory for Surgical Modeling Simulation and Robotics 4

Hypothesis: Resident surgeon drill motion patterns during dissection of a printed and cadaveric temporal bone model are anticipated to be dissimilar owing to material properties. Background: Virtual haptic and physical printed temporal bone simulations are commonly used to augment cadaveric training. Assessment of these tools is ongoing with a strong trainee preference for physical simulations. Trainees using virtual haptic models illustrate disparate drill motion patterns compared to cadaveric opportunities, which could result in maladaptive skill development. Methods: Resident surgeons dissected printed bones generated from microCT data and cadaveric specimens. Skill assessment was clustered into cortical mastoidectomy, thinning procedures (sigmoid sinus, dural plate, posterior canal wall) and development of a posterior tympanotomy. A magnetic position tracking system (TrakSTAR, Ascension) captured drill position and orientation at 200Hz. Dissection was performed by 8 trainees (n=5 < PGY3 > n=3) using kcos-metrics to analyze drill strokes within position recordings. Results: t-tests between models showed no significant difference in drill stroke frequency (cadaveric=1.36/s, printed =1.50/s p=0.420) but demonstrate significantly shorter duration (cadaveric=0.37s, printed =0.16s p<0.05) and a higher percentage of curved strokes (cadaveric= 31, printed =67 p<0.05) used in printed dissection procedures. Junior staff used a higher number of short strokes (junior = 0.54, senior =0.38, p<0.05) and higher percent of curved strokes (junior = 35%, senior =21%, p<0.05). Conclusion: Significant differences in hand motions were present between the cadaveric specimens and printed simulations, questioning the employ of printed simulations as viable teaching instruments. Junior staff appear to adopt a more cautious approach to dissection. Funding: Edward Ross Pressman Memorial Award, Department of Otolaryngology, H.T Thorlakson Foundation, HSC Research Foundation, College of Medicine Research Manitoba

Evaluating medical studentsâ&#x20AC;&#x2122; interest and readiness for learning and practicing high-value care: Preparing for change Noren Khamis1, Ahmad Sidiqi1, Malcolm Maclure1, Geoffrey Blair1 1

The University of British Columbia, Vancouver, British Columbia

Key words: high-value care, resource stewardship, medical education Rationale: The CanMEDS framework was recently updated to include high-value care and resource stewardship as a core competency. This principle, which encourages appropriate and judicious use of diagnostic and therapeutic actions, promotes patient safety and fosters financial sustainability of the health care system. We posit that if physicians learn the concepts of resource stewardship and high-value care early in their training, they will be empowered to reduce wasteful low-yield tests and potentially harmful treatments. However, whether current undergraduate and residency training programs effectively educate trainees on high-value care remains unknown. Objectives: We aim to test: 1) medical studentsâ&#x20AC;&#x2122; current knowledge around resource stewardship; 2) the importance students place on high-value care education; 3) the ideal format to train students on this subject. Results will guide appropriate curriculum adjustments. Methods: A validated and research ethics-approved 21-item online survey was piloted on 43 medical students and subsequently distributed to all medical students at the University of British Columbia via an electronic mailing list. The survey was voluntary. Student perspectives were analysed using descriptive statistics. Results: There were 88 survey responses. There were at least 15 responses from each cohort of medical students (years 1-4). The majority of students (93%) agreed or strongly agreed on the importance of resource stewardship in clinical decision-making; however, all respondents felt that their training has inadequately prepared them on this topic, and only 28% felt comfortable discussing costs of care and overtreatment with patients. Furthermore, most respondents (85%) believed that training in resource stewardship and appropriate care needs to begin in the first two years of medical school. Conclusions: UBC medical students value resource stewardship education, but their lack of adequate training on the subject prevents them from advocating for appropriate patient care and resource allocation. Increased emphasis on resource stewardship in the undergraduate medical curriculum, especially in the first two years of medical school, may help trainees subsequently make safe and cost-effective health care decisions. Further studies are needed to assess the effect of curricular change on helping trainees competently implement concepts of resource stewardship into their clinical practice.

Email: nkhamis@alumni.ubc.ca

Novel analyses of cognitive flexibility in a transgenic hAPP rat model with comorbid striatal ischemic stroke Aaron Regis1, Alexander Levit1, Vladimir Hachinski2, Brian Allman1, Shawn Whitehead1 1 Department of Anatomy & Cell Biology, Schulich School of Medicine & Dentistry, Western University, London, ON 2 Department of Clinical Neurological Sciences, University Hospital, Western University, London, ON Background: Early Alzheimer disease (AD) and striatal stroke are both thought to impair cognitive flexibility and a comorbidity is hypothesized to have synergistic effects. Cognitive inflexibility can be modeled in rodents, and we present new analyses of cognitive flexibility in a comorbid rat model. Animal Model & Methods: Transgenic Fischer 344 rats overexpress pathogenic human amyloid precursor protein (hAPP). Ischemic strokes were induced with unilateral endothelin (ET-1) injections in the striatum of TgF344 and wildtype control rats. Using an operant chamber, rats were challenged to shift from a formerly rewarding (visual cue) strategy to a new (lever location) strategy, a task that requires cognitive flexibility. Results: Current standards for analysis indicate that the TgF344 and the ET-1 injected rats committed more regressive errors (p<0.01, p<0.005); these effects were additive but not synergistic. The TgF344 rats also committed more never-reinforced errors (p<0.01). By also evaluating the rate at which the rats shift strategies, it was found that ET-1 injection significantly lengthened the transition period between old and new strategies (p<0.005), particularly in TgF344 rats (p<0.05). Conclusions: Trial dependent analyses offer added insight into rat cognitive flexibility during setshifting. Striatal stroke and pathogenic hAPP interact to produce synergistically worse cognitive outcomes.

Effectiveness of Interactive Medical Language Workshops for Helping Chinese-speaking Patients with Limited English Proficiency David Denga, Rayleigh Chana, Bo Gonga, Jeffrey Chana, Donna Liaoa, Kendall Hob a MD Undergraduate Program, University of British Columbia b Department of Emergency Medicine, Faculty of Medicine, University of British Columbia Rationale: Individuals of Chinese ancestry make up a significant portion of Canadaâ&#x20AC;&#x2122;s visible minority. Although research into the importance of language as a healthcare barrier remains limited in Canada, studies have shown that lack of language proficiency can result in worse patient health outcomes. Objective: To explore the healthcare experiences of Chinese-speaking patients with limited English proficiency living in Canada and to assesse the effectiveness of patient education for this clinical population through the delivery of customized, interactive medical English workshops. Methods: Native Chinese-speaking patients were recruited through the interCultural Online health Network, a University of British Columbia Department of Emergency Medicine initiative to support Chinese patients in chronic disease management through health education. A sample of respondents was recruited to participate in a focus group. Two moderators then explored specific language-related communication issues faced by these patients in accessing healthcare in Vancouver. The findings were used to inform the Medical English Workshop curriculum. The workshops were then delivered to native Chinese-speaking patients, and assessed through standardized surveys. Results: A cohort of 18 native Chinese-speakers (9 males, 9 females) participated in our workshops. The average age of participants was 67. Prior to the workshop, 39% and 53% of participants noted lack of English proficiency was a frequent or occasional barrier of care, respectively. Interestingly, all participants currently had family physicians who shared their native language. However, many expressed difficulties interacting with specialists, who tend not to speak Chinese. Approximately 56% of participants recalled adverse medical experiences, in part due to ineffective communication with healthcare providers. Post-workshop surveys revealed that 56% of participants found the session to be extremely helpful and all participants indicated that they would recommend similar workshops to others. Conclusion: This study validated that Chinese patients in Vancouver experience similar barriers of care due to language as reported in the literature, and such barriers seemed higher when accessing specialty care. Language workshops designed for patients to overcome such barriers were favourably received. Future studies should be done to examine if this approach can be a cost-effective way of empowering patients with limited English proficiency. Funding: None Email: Donna.l@alumni.ubc.ca

Fibre Intake Following Exclusive Enteral Nutrition (EEN) Therapy Alters Microbiome in Pediatric Crohn’s Disease Patients Amber L. MacLellana, Jessica Moore-Connorsb, Brad MacIntyreb, Gavin Douglasc, Katherine A. Dunnd, Joseph P. Bielawskid, Angela Nobleb, Gamal Mahdib, Mohsin Rashidb, Anthony Otleyb, Leah E. Cahilla,e, Morgan G.I. Langillec, and Johan Van Limbergena,b. a

Department of Medicine, Dalhousie University, Halifax, Canada Division of Pediatric Gastroenterology & Nutrition, Department of Pediatrics, IWK Health Centre, Dalhousie University Halifax, Canada c Department of Pharmacology, Dalhousie University, Halifax, Canada d Department of Biology, Dalhousie University, Halifax, Canada e Department of Community Health and Epidemiology, Dalhousie University, Halifax, Canada b

Rationale: Changes in gut microbiome community structure are associated with the development of Crohn’s Disease (CD) and can be altered by environmental factors such as diet. Exclusive enteral nutrition (EEN) can induce disease remission in more than 80% of pediatric CD patients. However, the mechanism of EEN effectiveness remains unclear and evidence-based dietary recommendations for remission maintenance following EEN cessation do not exist. Objectives: We aimed to assess the influence of dietary fibre intake on microbiome composition and sustained remission (SR) in pediatric CD patients who return to their regular diet post-EEN. Methods: A 79-item validated food frequency questionnaire (FFQ) was used in cross-sectional analysis of dietary fibre intake for 11 pediatric participants (11-17 years) in an ongoing study of EENinduced gut microbiome changes. Participants had returned to a regular ad libitum diet for >3 months following 12-week EEN remission induction therapy. QIIME was used to analyze 16S rRNA sequences from participant stool samples, collected at 12-week intervals, and assess corresponding changes in microbiome diversity. Results: Due to low fibre intake, participants were classified as ‘higher fibre’ if they consumed at least half of the daily age and sex-matched fibre recommendations, and ‘lower fibre’ if they did not. Microbial beta diversity was significantly lower in patients consuming lower fibre vs. higher fibre (p = 0.04, ANOSIM). Microbiome diversity was not significantly different for patients consuming < 600g of oral solid food (n = 3) vs. those consuming >600g. Chao-1 alpha diversity analysis showed lowest gut microbial richness in patients who achieved remission with EEN but experienced disease flare by 6-month follow-up with regular diet (wPCDAI >12.5, n = 3), compared to EEN primary non-responders (n = 2), and those in SR (n = 4). Conclusion: Lower fibre intake is associated with decreased gut microbial diversity in pediatric CD patients following induction treatment with EEN. Persistence of decreased gut microbiome diversity with return to regular diet post-EEN may indicate increased risk of disease flare. Further longitudinal analysis of microbiome changes associated with return to oral diet after EEN are required to inform dietary therapy recommendations for remission maintenance in pediatric CD. Funding: NASPHGAN, CIHR, NSHRF, DMRF, Crohn’s & Colitis FoA, FoM Webster Studentship Email: amber.maclellan@dal.ca 48

Genetic Diversity and Recombination in Circulating Serotypes from the Enterovirus B Species Andrew McDermid (1,2), Elsie Grudeski (2), Michelle Gusdal (2), Russell Mandes (2) and Tim Booth (1,2) 1. University of Manitoba, Department of Medical Microbiology 2. National Microbiology Laboratory, Winnipeg, Manitoba Introduction: Enteroviruses are the leading cause of viral meningitis. Other notable diseases associated with Enteroviruses include hand, foot and mouth disease, poliomyelitis, type I diabetes, myocarditis, and encephalitis. Enteroviruses infecting humans can be classified into 7 species, and further into hundreds of serotypes. Genetic variation in Enteroviruses can occur by recombination when two viruses within the same species coinfect a cell and exchange genetic material or by mutation, though rates and limits to this variation are not completely understood. RNA Dependant RNA Polymerase lacking proofreading leading to very high mutation rates. Treatment and prevention of Enterovirus disease is a challenge due to the high genetic variation. The Enterovirus B species is the most prevalent species isolated by Canadian surveillance. Enterovirus B most notably includes all Echoviruses and many Coxsackieviruses. Here we examine recent Enterovirus B strains that represent the whole species in order to better understand genetic variation. Methods: Two Canadian Enterovirus Surveillance systems, the National Centre for Enteroviruses (NCEV) and the Canadian Virus Report (CVR), were used to determine the most frequently circulating Enterovirus serotypes from 1991-2014. Representative isolates from selected serotypes were selected from the NCEV repository for full genome sequencing. More than 160 isolates from 17 serotypes representing 93.4% of the surveyed Canadian Enterovirus B gene pool and more than 20 years of surveillance were fully sequenced. Recombination events were identified using nucleotide identities in defined gene regions of the samples. Recombination events were analysed within serotypes. Results: Surveillance data show dozens of serotype-specific outbreaks in Canada over the past two decades. Recombination events were often closely associated with these outbreaks. Evidence suggests evolution of different portions of the genome is independent. Conclusions: Identification of conserved Enterovirus sites exclusively in outbreak strains can provide better understanding of circulating Enteroviruses and viral evolution. We anticipate this study will be valuable in identifying and defining limits to genetic niches within Enterovirus species. Such limits could be exploited when choosing targets for therapeutics.

Slit2 Decreases the Size of Atherosclerotic Lesions in mice by Preventing Macrophage Oxldl Uptake and Foam Cell Formation Ilya Mukovozov1,2, YW Huang1, GY Liu1, S Patel1, HS Wong1,2, and LA Robinson1,2 1 Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, ON 2 Institute of Medical Science, Faculty of Medicine, University of Toronto, Toronto, ON Background: Atherosclerosis is a maladaptive response initiated by the retention of cholesterol-rich low-density lipoproteins, which are rendered pro-inflammatory upon oxidation. While the clearance of oxidized low-density lipoprotein (oxLDL) by macrophages is initially beneficial, excessive accumulation results in cholesterol-laden foam cells that drive disease pathogenesis resulting in the growth of atherosclerotic lesions. The uptake of oxLDL is mediated predominantly by the scavenger receptor CD36 and requires dynamic actin reorganization driven by Rac GTPases. However, external signals that regulate this process remain elusive. The Slit family of secreted proteins, together with their transmembrane receptor Robo, inhibit the migration of a variety of cell types, in part by inactivating Rac and blocking dynamic actin rearrangement. Thus, it is conceivable that Slit-Robo signaling may be harnessed to prevent the progression of atherosclerosis by limiting the uptake of oxLDL and macrophage foam cell formation. Purpose: To investigate the role of Slit2 in atherosclerotic lesion development in ldlr−/− mice and in oxLDL-mediated foam cell formation. Hypothesis: We hypothesize that Slit2 will decrease the size of atherosclerotic lesions and blunt oxLDL-mediated foam cell formation in macrophages by blocking Rac dependent uptake of oxLDL by CD36. Methods: To measure atherogenesis in vivo, ldlr−/− mice were placed on a cholesterol rich diet (CRD) for 6 weeks, then aortas were harvested and mounted en face following perfusion fixation. In parallel experiments, isolated aortas were enzymatically digested and cells analyzed by flow cytometry. Peripheral blood mononuclear cells were isolated by density centrifugation and monocytes polarized for 7 days to macrophages. Foam cell formation was investigated using a neutral lipid probe, BODIPY 497/503, and quantified by immunofluorescence microscopy and flow cytometry. The binding and uptake of oxLDL was investigated using fluorescently (DiI)-labeled oxLDL, and quantified by microscopy and flow cytometry. Results: Exogenous Slit2 decreased atherosclerotic lesion size in ldlr−/− mice placed on a 6 wk CRD. Human and murine macrophages express the Slit2 receptor, Robo-1. Treatment with Slit2 inhibited foam cell formation in human and murine macrophages (P<0.01). Slit2 blocked macrophage binding and uptake of oxLDL (P<0.01) and inhibited oxLDL-mediated activation of Rac1. Conclusions: Slit2 decreases the size of atherosclerotic lesions in ldlr−/− mice and inhibits oxLDL-mediated foam cell formation in both human and murine macrophages. Funding: CIHR MD/PhD Email: ilya.mukovozov@utoronto.ca

Antenatal suppression of interleukin-1 protects against inflammation-induced fetal injury and improves neonatal and developmental outcomes in mice Mathieu Nadeau-Vallée1, Peck-Yin Chin2, Lydia Belarbi1, Marie-Ève Brien3, Sheetal Pundir1, Alexandra Beaudry-Richard1, David J Sharkey2, Xin Hou1, Christiane Quiniou1, Jean-Sébastien Joyal1, William D. Lubell4, David M. Olson5, Sarah A. Robertson2, Sylvie Girard3 and Sylvain Chemtob1. 1

Departments of Pediatrics, Ophtalmology and Pharmacology, CHU Sainte-Justine Research Center, Montreal, Qc, Canada; 2Department of Obstetrics and Gynecology, Adelaide Medical School and Robinson Research Institute, University of Adelaide, Adelaide, SA, Australia; 3 Departments of Obstetrics and Gynecology, and Microbiology, Infectiology and Immunology, CHU Sainte-Justine Research Center, Montreal, Qc, Canada; 4Department of Chemistry, Université de Montréal, Montréal, Qc, Canada and 5Departments of Obstetrics and Gynecology, Pediatrics and Physiology, University of Alberta, Edmonton, Ab, Canada. Rationale: Preterm birth (PTB) is commonly accompanied by in utero fetal inflammation, and existing tocolytic drugs do not target fetal inflammatory injury. Of the candidate proinflammatory mediators, interleukin-1 (IL-1) appears central and sufficient to trigger fetal loss. Methods: Therefore, we elucidated the effects of antenatal IL-1 exposure on post-natal development, and investigated two IL-1 receptor antagonists, the competitive inhibitor Kineret (anakinra) and a potent noncompetitive inhibitor 101.10, for efficacy in blocking IL-1 actions. Results: Antenatal exposure to IL-1β induced Tnfa, Il6, Ccl2 and Pghs2 expression in placenta and fetal membranes, and elevated amniotic fluid IL-1β, IL-6, IL-8 and PGF2α, resulting in PTB and marked neonatal mortality. Surviving neonates had increased Il1b, Il6, Il8, Il10, Pghs2 and Tnfa expression in white blood cells with elevated plasma IL-1β, IL-6 and IL-8, increased IL-1β, IL-6 and IL-8 in fetal lung, intestine and brain, and morphological abnormalities including disrupted lung alveolarization, atrophy of intestinal villus and colon-resident lymphoid follicle, and brain microvascular degeneration and atrophy with visual evoked potential anomalies. Late gestation treatment with 101.10 abolished these adverse outcomes, whereas Kineret exerted only modest effects and no benefit for gestation length, neonatal mortality or placental inflammation. In an LPS-induced model of infection-associated PTB, 101.10 prevented PTB, neonatal mortality and fetal brain inflammation. Conclusions: The results implicate IL-1 as an important driver of neonatal morbidity in PTB and identify 101.10 as an effective candidate therapeutic to counteract its harmful actions. Funding: CIHR, Global Alliance for the Prevention of Prematurity and Stillbirth, SickKids Foundation Email: mathieu.nadeau-vallee@umontreal.ca

Validation of Microfluidic Encapsulation of Cardiac Stem Cells for Myocardial Repair. Alex Nantsiosa, A Benaventeb, P Kandaa, and Dr. DR Davisa a b

The University of Ottawa Heart Institute, Ottawa, ON, CANADA The University of Ottawa, Department of Physics, Ottawa, ON, CANADA

Rationale: Ischemic cardiomyopathy, the most common form of heart failure, affects over 600,000 Canadians. Despite pharmacologic innovations in cardiac care, the 1-year mortality remains high at 25%. To address this challenge, we have developed techniques to grow cardiac stem cells (CSCs) from small myocardial biopsies for delivery to areas of injury, where they engraft and repair myocardium. CSCs possess capacity to regenerate heart tissue, previously shown to be capable of differentiation into all cardiac cell lineages, secretion of cardioprotective cytokines, and to improve cardiac function in mice post-MI. Due to their low rate of engraftment, we have developed a clinically translatable technique to encapsulate the CSCs before injection, into an agarose cocoon supplemented with fibrinogen and fibronectin, using vortex based emulsions. However, this vortex-based-technique offers little control over capsule size and uniformity. We have developed a microfluidics-based approach, which encapsulates cells in a supportive cocoon at a high-throughput rate, offering control over size and occupancy, while limiting the mechanical stress. Objectives: To evaluate the influence of microfluidic encapsulation on the viability, extracapsular migration and paracrine secretion of human CSCs. Methods: Humans CSCs were cultured from atrial appendages from patients undergoing cardiac surgery. CSCs were mixed with low melt agarose, and encapsulated in two ways i) standard vortex based encapsulation, ii) microfluidic encapsulation. Viability and proliferation of cells were assessed with CCK- 8 assay and confirmed with LIVE-DEAD staining. Extracapsular migration was measured by simple cell counts. Results: Microfluidic encapsulation provides more uniform cell capsule sizes, a more constant occupancy, reducing batch to batch variation. Preliminary results indicated that microfluidic encapsulation reduces viability of the CSCs compared to standard encapsulation. After pre-coating the device with BSA, however, there was no significant difference in proliferation of the CSC encapsulated with either method and improved viability was observed in the microfluidic method compared to standard method (p < 0.001). Pre-coating the device, drastically reduces clogging and improves yield Conclusions: Cardiac stem cells have potential to regenerate the myocardium and improve ejection fraction post-MI, especially when encapsulated. Although requiring optimization, early data shows that microfluidic encapsulation has potential to be just as effective method of delivery of these cells into the heart, compared to the standard vortex-based method. Funding: Heart & Stroke Foundation Email: anant019@uottawa.ca

Non-CAG triplet repeat expansion mutations in ataxin-7 and top1mt lead to ataxia Siddharth Natha, Nicholas Caronb, Mark Tarnopolskyc, and Ray Truanta a

Department of Biochemistry and Biomedical Sciences, Michael G. DeGroote School of Medicine, Faculty of Health Sciences, McMaster University, Hamilton, Canada, bCentre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, Canada, c Division of Neurology, Department of Medicine and Department of Pediatrics, Michael G. DeGroote School of Medicine, Faculty of Health Sciences, McMaster University, Hamilton, Canada Rationale: Ataxin-7 is a multifunctional protein that has been implicated in the pathogenesis of spinocerebellar ataxia type 7 (SCA7), a fatal CAG triplet repeat disorder characterized by progressive cerebellar and retinal degeneration. Pathology in SCA7 results from a CAG triplet repeat expansion in excess of 37 in the first exon of the ATXN7 gene, leading to a polyglutamine tract expansion in its protein product, ataxin-7. Previously, through fluorescencelifetime imaging microscopy to measure Förster resonance energy transfer (FLIM-FRET), our group has shown, in the context of Huntington’s disease, that pathogenic polyglutamine tract expansion leads to a gain of structure, potentially limiting intermolecular and intramolecular interactions. Here, we present a novel SCA variant, manifesting clinically in a pediatric patient as severe ataxia in the absence of retinal degeneration as a consequence of Q35P and N556K mutations within an un-expanded (10 glutamines) ataxin-7, in concert with mutations in the nuclear-encoded mitochondrial topoisomerase, top1mt, as delineated by whole exome sequencing. Objectives: Determine how the novel mutations observed in our patient alter the structure of ataxin-7 and top1mt, and how these affect cellular biology. Methods: Skin fibroblasts were sampled from our patient, his parents, a healthy individual, and an individual with SCA7. Molecular cloning approaches were implemented to generate a FRET sensor, and FLIM-FRET was used to observe changes in protein structure. Immunofluorescence, western blotting, and fluorescence microscopy were used to study cellular biology within sampled cells. Seahorse metabolic analysis was done in order to assess mitochondrial function across cell types. Results: We observe that patient cells have increased nuclear localization of mutant ataxin-7, beyond what is seen in SCA7, as well as altered mitochondrial morphology and function. Furthermore, using FLIM-FRET, we demonstrate that the Q35P mutation within ataxin-7 drastically alters the protein’s structure, more so than a polyglutamine tract expansion. We also observe increased nuclear localization of ataxin-7 in wildtype human cells in response to mitochondrial stress, suggesting a link between ataxin-7 and the mitochondria. Conclusions: Ataxin-7 has established roles as a component of transcriptional machinery in the nucleus, and as a microtubule stabilizer in the cytosol. We propose that there also exist interactions between ataxin-7 and the mitochondria, and that loss of these interactions, may contribute to pathology in our novel SCA variant, and potentially, in SCA7. This work not only expands our understanding of ataxin-7 biology, but also provides insight into the cellular and molecular mechanisms underlying cerebellar neurodegeneration. Funding: CIHR, CFI, CHDI Foundation Inc., Huntington Society of Canada, Janes Family Foundation, Krembil Foundation 53

Mutation in Ubiquilin-2 exacerbates ALS/FTD features in a TDP-43 mouse model Vincent Picher-Martel1, Karine Plourde1, Christine Bareil1 and Jean-Pierre Julien1 1

Research Centre of Institut Universitaire en Santé Mentale de Québec and Department of Psychiatry and Neuroscience, Laval University, 2601 Chemin de la Canardière, Québec, QC, G1J 2G3, Canada Rationale: Mutations in the gene encoding Ubiquilin-2 (UBQLN2) are linked to amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). UBQLN2 plays a central role in ubiquitin proteasome system (UPS) and UBQLN2 mutants can form cytoplasmic aggregates in vitro and in vivo. We have recently demonstrated that UBQLN2 aggregates are dynamic structures that promote cytoplasmic accumulation of TAR DNA-binding protein (TDP-43), a major component of ALS inclusion bodies in neurons (Picher-Martel V. et al., 2015). Objectives: There is an important lack of conclusive animal models in ALS research. Indeed, the aim of the present study is to analyze whereas mutations in both UBQLN2 and TDP-43 genes can increase cytosolic inclusion bodies, lead to important motor and cognitive impairment in mice and mimic human ALS/FTD. Methods: For this purpose, we have generated new transgenic UBQLN2P497H mice by microinjection of flag-tagged UBQLN2 gene under control of NFH promoter in one-cell C57BL6 zygotes. UBQLN2P497H mice were then bred with our well characterized and acknowledged TDP-43G348C mice model previously described in Swarup V. et al., 2011 to generates double transgenic UBQLN2P497H; TDP-43G348C mice. Results: Our double transgenic UBQLN2P497H; TDP-43G348C mice developed important cognitive deficits at passive avoidance test at 7 months of age. They exhibit an increase TDP-43 proteinopathy in spinal cord and brain at the age of 5 months. They also developed muscle atrophy and motor phenotype at a faster rate than the simple transgenic TDP-43G348C mice. Conclusions: Double transgenic UBQLN2P497H; TDP-43G348C mice develop typical features of ALS/FTD and could be exploited as a better animal model to test therapeutic avenue. Funding: CIHR, FRQS Email: Vincent.picher-martel.1@ulaval.ca

Life-Threatening Motor Vehicle Crashes Under Bright Sunlight Sheharyar Raza [1,2] and Donald Redelmeier [1,2,3] [1] Faculty of Medicine, University of Toronto [2] Institute of Clinical Evaluative Sciences (ICES) in Ontario [3] Institute for Health Policy Management and Evaluation Rationale: Bright sunlight may create visual illusions from aerial perspective or other sources of driver error. Objective: We tested whether the risk of a life-threatening motor vehicle crash is increased when driving under bright sunlight. Methods: We conducted a case-only longitudinal analysis of consecutive patients hospitalized in Canada's largest trauma center from January 1, 1995 to December 31, 2014 because of a motor vehicle crash. The prevailing weather at the time and place of the crash was compared to the same hour and location on control days a week earlier and a week later. Results: Overall, the majority of patients (n = 6,962) were injured during daylight hours and bright sunlight was the most common weather condition at the time and place of the crash. The risk of a life-threatening crash was 16% higher during bright sunlight compared to controls (95% confidence interval: 9 to 24, p < 0.001). The increased risk was accentuated in the early afternoon, disappeared at night, extended to patients with different characteristics, involved crashes with diverse features, and not apparent with cloudy weather. The increased risk extended to patients with high crash severity as indicated by ambulance involvement, length of hospital stay, intensive care unit admission, or patient mortality. Conclusions: Bright sunlight is associated with an increased risk of a life- threatening motor vehicle crash. An awareness of this risk might inform behavior change, safety policy, and physician warnings to prevent a life-threatening motor vehicle crash. Funding: Comprehensive Research Experience for Medical Students at the University of Toronto Canada Research Chairs Grant Canadian Institutes of Health Research Email: s.raza@utoronto.ca

Systemic Inflammation during antiretroviral therapy initiation as a predictor of immune response among HIV-infected individuals in Manitoba Quinlan Richert,a Adriana Trajtman,a Luisa Arroyaved, Julia Toews, Marissa Beckerb, Ken Kaspera,b, Zulma Ruedad and Yoav Keynana,b,c,* a. Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada b. Department of Internal Medicine, University of Manitoba, Winnipeg, Canada c. Community Health Sciences, University of Manitoba, Winnipeg, Canada d. Facultad de Medicina, Universidad Pontificia Bolivariana, Medellin, Colombia

Introduction: Despite the life-prolonging effects of Highly Active Antiretroviral Therapy (HAART), persons with HIV are still prone to higher rates of non-AIDS related morbidity (such as heart, kidney, and liver disease) than the general public. This is likely due to chronic immune activation and inflammation that persists in HIV-positive persons despite virological suppression. What remains undetermined, however, is whether a link exists between chronic inflammation/immune activation and suboptimal immune recovery on HAART. The hypothesis of the present study is that higher levels of systemic subclinical inflammation and immune activation are linked with suboptimal immune recovery on HAART. Methods: Fifteen eligible patients from the Manitoba HIV program were enrolled and followed for up to two years; blood samples were drawn at 4 timepoints each, and concentrations of 21 proinflammatory markers were measured. Patients were grouped according to CD4:CD8 recovery at viral suppression, and the inflammatory profiles of the two groups were compared. Results and conclusions: APRIL and BAFF are higher in those with poor recovery at the point of viral suppression, but were also higher in this group at the onset of therapy and through the three additional follow-up visits. TNF-R1, CD163, and Osteopontin, were also in higher concentrations at the outset of therapy and beyond. These five molecules could thus see potential use in the future as biomarkers of likely poor immune recovery. Future work should focus on replicating these findings with larger cohorts.

Infertility and womenâ&#x20AC;&#x2122;s mental health: The relationship of female-factor infertility and depression and anxiety Nadine Rockwooda, Amanda Pendergastb, and Deanna Murphyc a

MD Candidate, Memorial University of Newfoundland, bDepartment of Family Medicine, Memorial University of Newfoundland, cDepartment of Obstetrics and Gynecology, Memorial University of Newfoundland. Newfoundland, Canada Rationale: Infertility, which affects between 11.5% and 15.7% of Canadian couples, may be attributed to female factors if women of reproductive age are unable to conceive within 12 months of unprotected intercourse. Infertility is described as a devastating experience, which can lead to psychological distress. While comprehensive, population based research initiatives exist surrounding infertility and mental health in Europe and Asia, there are no published studies in the Canadian population, including Newfoundlanders and Labradoreans. It is crucial to identify the risk of developing depression/anxiety symptoms, so these women can be identified and provided appropriate resources for their mental health and well-being. Objectives: The objectives of this study were three-fold: to assess the relationship between infertility and development of depression/anxiety symptoms among women referred to the fertility clinic using the Beck Depression Inventory II (BDI) and the Beck Anxiety Inventory (BAI); to compare infertile womenâ&#x20AC;&#x2122;s BDI/BDA to the scores of pregnant infertile and fertile counterparts; to study the influence of demographic and other factors on depression and anxiety scores after receiving an infertility diagnosis. Methods: Female patients attending a fertility clinic and a family practice clinic were asked to participate in a mixed methods survey assessing depression/anxiety symptoms, details of infertility, BDI/BDA scores, and experiences with infertility. The diagnosis of female-factor infertility and depression/ anxiety scores will be examined and compared to depression/anxiety scores of pregnant infertility and non-infertility patients. Results: We hypothesized that women with an infertility diagnosis will exhibit greater BDI/BDA scores compared to women who are currently pregnant or not experiencing fertility issues, with duration of infertility exacerbating levels of depression and anxiety symptoms. Preliminary data will be presented. Conclusions: Based on the findings from this study, conclusions will be drawn regarding the relationship of infertility and depression and anxiety as well as potential demographic variables and risk factors that may influence this experience in a Newfoundland and Labrador population. Identifying at risk individuals is paramount, as protocols and polices can be put in place to provide these women with appropriate resources to improve their mental health and well-being. Email: nmer67@mun.ca

The role of EZH2 in KRASG12D-mediated acinar-to-ductal metaplasia Ryan JR1,2, Johnson CL2, Berger K2, Urrutia, R.4, Lomberk, G.4, and Pin CL1-3 1 Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada Children’s Health Research Institute, London, ON, Canada

2 3

Departments of Paediatrics, Physiology and Pharmacology, and Oncology, University of Western Ontario, London, ON, Canada 4

Mayo Clinic, Rochester, MN, USA

Rationale: Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer-related death in North America. Activating KRAS mutations (e.g. KRASG12D) play a key role in PDAC pathogenesis. Tumor genetic analysis has shown elevated expression of histone methyltransferase Enhancer of Zeste Homologue 2 (EZH2) correlates with poorer prognosis. Further characterization of EZH2’s role in KRASG12D-mediated PDAC initiation and progression is critical for understanding the disease process and the development of novel therapeutic agents in PDAC. Objectives: (1) Investigate EZH2 expression in normal human pancreatic tissue and in the context of mild or severe tissue injury. (2) Determine the effects of oncogenic KRASG12D in acinar cells in the presence and absence of EZH2. Methods: Expression of Ezh2 and acinar and duct cell differentiation markers was assessed by RT-PCR analysis in human pancreatic tissue samples from PDAC patients and in human pancreatic cancer and normal acinar cell lines. Tissue morphology and expression of cellular differentiation markers was also assessed in a mouse model for inducible KRASG12D expression and or deletion of the Ezh2 SET domain (LSL-KRASG12DEzh2flSET). Histologic and qRT-PCR analyses were performed on wild type, Ezh2-/-, KRASG12D, and LSL-KRASG12DEzh2flSET mice. Results: Ezh2 expression positively correlated with the severity of tissue injury and loss of mature acinar cell markers in human tissue. LSL-KRASG12D mice showed loss of mature acinar cell marker Nr5a2 with increased expression of progenitor cell markers Rbpj and Sox9 and ductal cell marker Krt19 in the presence of EZH2. Conclusions: High Ezh2 expression in pancreatic cancer patients and LSL-KRASG12D mice is coupled with the loss of mature acinar cell markers suggests EZH2 mediates the process of acinar to ductal cell metaplasia in the presence of active KRAS. Investigation of the direct gene targets of EZH2 in the context of pancreatic acinar cell injury should be conducted to further characterize the role of EHZ2 in early PDAC pathogenesis. Funding: Children’s Health Research Institute, CIHR, Cancer Research Society, Rob Lutterman Foundation, and Mach-Gnaslsson Foundation.

Investigating ischemia-reperfusion (I/R) injury: Identification of a novel inflammatory axis induced by hypoxia in primary endothelial cells. Nicholas Sajko1, Ian Alwayn1,2, and Jennifer Corcoran1,2 1

Department of Microbiology & Immunology, 2Department of Surgery, Dalhousie University, Halifax, NS, Canada. Rationale: One of the main underlying causes of liver transplantation failure is ischemia/reperfusion (I/R) injury. This biphasic process involves blood and oxygen deprivation, beginning after the graft is removed from its original host blood supply. The associated hypoxic conditions disrupt many life-sustaining processes, particularly affecting the liver sinusoidal endothelial cells (LSECs). Under these conditions, LSECs undergo cell deregulation and cytoskeletal changes leading to poor endothelial cell (EC) barrier function and the formation of an inflammatory phenotype, contributing to subsequent hepatocyte damage. Currently, the precise molecular mechanisms underlying this relationship are poorly understood. Recent studies point to the potential involvement of a stress-responsive MAP kinase signaling cascade, the MK2-RhoA signaling axis, which has been linked to controlling EC pro-inflammatory cytokine expression and barrier function. Objectives: Determine the potential involvement of the MK2-RhoA signaling axis in EC dysregulation and inflammation associated with the hypoxic conditions seen in settings of I/R injury. Methods: Investigations were done using an in-vitro system of I/R associated hypoxia and a Human umbilical vein EC (HUVEC) model. Immunoblotting was performed to detect the activation of various constituents of the MK2-RhoA pathway. Immunofluorescence was used to monitor general cytokine expression via detection of cellular processing bodies (PBs); cytoplasmic ribonucleoprotein mRNA decay granules that function in aggregate to limit expression of pro-inflammatory cytokines and whose disassembly is linked to the MK2-RhoA axis. Results: Both hypoxic (1% O2) and anoxic (0% O2) conditions activate the MK2-RhoA axis, as demonstrated by the increased phosphorylation of MK2 and its downstream target, Hsp27. Additionally, we observed actin stress fiber formation under hypoxic conditions consistent with those observed in MK2-RhoA activation and I/R injury. Preliminary data also shows PB disassembly under hypoxia as compared to no treatment controls, suggesting an increased expression of pro-inflammatory cytokines. Conclusions: The results form a link between hypoxia and the activation of the MK2-RhoA axis in an EC model, and may suggest that deregulated PB homeostasis is a central feature of ECinduced inflammation during I/R injury. Further work aims to elucidate specific changes in proinflammatory cytokine expression in HUVECs exposed to hypoxia, and subsequently move our model into LSECs. Funding: Dalhousie Medical Research Foundation (DMRF); The Dalhousie Surgery SEED Funding Program; The JJ Carroll Travel Fund. Email: nick.sajko@dal.ca 59

Reflex testing for germline BRCA1, BRCA2, PALB2 and ATM mutations in pancreatic cancer Alyssa L. Smitha,b, Cavin Wonga,b, Adeline Cuggiaa,b,c, Ayelet Borgidad, Spring Holterd, Anita Halla.b, George Chonge, Mohammad R. Akbarif, William D. Foulkesa,c, Steven Gallingerd, George Zogopoulosa,b,c a

Cancer Research Program, Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada, bGoodman Cancer Research Centre, McGill University, Montreal, Quebec, Canada, cProgram in Cancer Genetics, Departments of Oncology and Human Genetics, McGill University, Montreal, Quebec, Canada, dLunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital. Toronto, Ontario, Canada, eMolecular Pathology Unit, Department of Pathology, Jewish General Hospital, Montreal, Quebec, Canada, fWomen’s College Research Institute, Women’s College Hospital, University of Toronto, Toronto, Ontario, Canada RATIONAL: Pancreatic adenocarcinoma (PAC) associated with defective homology-directed DNA repair (HDR), frequently attributable to inherited germline mutations in HDR genes, has been shown to have increased therapeutic sensitivities to platinum-based DNA cross-linking agents and poly(ADP-ribose) polymerase inhibitors. Detecting mutation carriers at the time of PAC diagnosis may guide therapeutic decisions and may translate to improved clinical outcomes for patients with this deadly malignancy. OBJECTIVES: We investigated whether reflex testing for germline mutations in 4 HDR predisposition genes should be offered to incident cases with PAC. METHODS: Consecutive patients with PAC and French-Canadian (FC) ancestry were evaluated for 20 FC founder (n=132) and non-founder (n=114) mutations in ATM, BRCA1, BRCA2 and PALB2. Prospective PAC cases (n=236) unselected for ancestry were also assessed by full gene sequencing for mutations in these genes. Overall survival (OS) of carriers versus non-carriers was compared. RESULTS: Eight of 114 (7.0%) FC cases carried mutations in BRCA2 (4.4%), PALB2 (1.8%) or ATM (0.9%). Of the 132 FC cases tested for founder mutations, 6 were carriers, for a FC founder mutation prevalence of 4.5%. Three of 30 (10.0%) cases with Ashkenazi Jewish (AJ) ancestry had founder mutations, while 10 of 206 cases (4.9%) without founder ancestry carried mutations in BRCA1 (0.5%), BRCA2 (2.9%) or ATM (1.5%). Nearly 1 in 6 patients diagnosed ≤50 years of age (P = .03) were carriers. Mutations were also more frequent in patients with 2 first- or seconddegree relatives with pancreas, breast, ovarian or prostate cancer (or 1 relative and a personal history of 2 of these cancers) (P < .0001). BRCA1, BRCA2 and PALB2 carriers with late stage (III/IV) disease had an OS advantage (P=.049), particularly if treated with platinums (P=.030). CONCLUSION: Reflex testing of incident PAC cases for germline mutations in these 4 HDR genes may become justified as the costs of full gene sequencing decline. In the interim, we recommend reflex founder mutation testing of patients with FC or AJ ancestry and full gene sequencing of patients who are ≤50 years or who meet the family history criteria described. Funding: CIHR (MD-PhD funding), Cedars Cancer Institute Fellowship, CRS, CCSRI Email: alyssa.smith@mail.mcgill.ca 60

MRI 3-Dimensional Texture Changes in the Motor Cortex Parallel Phenotypic Heterogeneity in Amyotrophic Lateral Sclerosis Naveet Sohi1, Yee-Hong Yang2, Sanjay Kalra1-4 1

Faculty of Medicine and Dentistry; 2Department of Computing Science ; 3Neuroscience and Mental Health Institute; 4Department of Medicine, Division of Neurology; University of Alberta, Edmonton, Alberta, Canada Rationale: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of the human motor system, with an average survival of 3-5 years from the date of clinical diagnosis. Difficulties in diagnosis, clinical management and drug trials are, in part, a result of extensive clinical heterogeneity in: the site of onset, rate of progression and lack of an objective measures of upper motor neurones. In pursuit of an objective biomarker, many advanced MRI structural modalities have failed to consistently translate ALS phenotypic heterogeneity and disease progression into expected patterns of focal grey matter pathology. Our, recently developed, MRI 3D texture analysis technique has shown significant sensitivity and specificity to structural, and longitudinal, textural change on MRI in ALS. Objectives: Determine the utility of MRI 3D texture analysis as an imaging biomarker to characterize focal changes in the motor strip between patients with limb and bulbar onset, and identify regions that correlate with the degree of clinical disability. Methods: 24 patients with ALS were recruited, scanned and assessed clinically utilizing the ALS Functional Rating Scale-Revised (ALS-FRS-R). MRI 3D texture analysis was utilized to quantify 5 textural values in the primary motor areas and phenotype specific regions of the motor homunculus. Subgroup, correlation and longitudinal analysis was done using SPM8 software. Results: Patients with bulbar onset weakness revealed significant textural differences (p < 0.05, FWE corrected) in the bulbar segment of the motor homunculus compared to the limb segment; conversely, limb onset patients revealed focal textural changes in the limb segment (p < 0.05, FWE corrected). Textural values in the limb segment correlated positively to functional ALSFRS-R scores (p < 0.001), both subgroups revealed diffuse textural changes throughout the motor strip longitudinally (p < 0.001). Conclusions: Our results suggest that 3D MRI texture analysis is a superior biomarker for evaluating phenotypic differences and monitoring progressive changes in the upper motor neurones of ALS patients, when compared to existing structural imaging modalities. The pathophysiological correlation implicated in textural changes remains under study. Funding: David and Beatrice Reidford Research Scholarship, ALS Society of Canada, ALS Association.

Total Arsenic and Speciation of Arsenic in Alberta Freshwater Fish Xiao Jing (Lily) Sun, Chuxuan Li, Xiufen Lu and X. Chris Le Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada T6G 2G3 Introduction: Different chemical forms of arsenic (As) exist naturally in the environment. The toxicity of As depends on its chemical form. Although much research has focused on arsenic species in seafood, little is known about the concentration of arsenic species in freshwater fish. The primary objective of this research is to determine the concentration of various arsenic species in three main types of fish collected from Alberta lakes. Methods: Arsenic species extracted from fish were separated by high-performance liquid chromatography and the concentrations of arsenic species were detected by inductively coupled plasma mass spectrometry. Results: Arsenobetaine (AsB), arsenate [As(V)], arsenite [As(III)], monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA) were identified and detected in fish samples. Walleye (Sander vitreus) from Lac la Biche (LC), Burnstick Lake (BK), Muskwa Lake (MW) and Skeleton Lake (SK) were found to have AsB (0.0011 to 0.055 µg/g), DMA (<0.00025 to 0.074 µg/g), MMA (<0.00025 to 0.0049 µg/g), As(V) (<0.00025 to 0.091 µg/g), and As(III) (<0.00025 to 0.00028 µg/g). Northern pike (Esox lucius) from LC, BK, MW, SK, Berry Creek Reservior (BY), North Buck Lake (NB) and Orloff Lake (OF) were found to have AsB (0.0012 to 0.080 µg/g), DMA (<0.00025 to 0.15 µg/g), MMA (<0.00025 to 0.0074 µg/g), As(V) (<0.00025 to 0.0091 µg/g), and As(III) (<0.00025 to 0.0074 µg/g). Lake whitefish (Coregonus clupeiformis) from NB and OF were found to have AsB (<0.00025 – 0.022 µg/g), DMA (<0.00025 – 0.00087 µg/g), MMA (<0.00025 µg/g), As(V) (<0.00025 to 0.0082 µg/g), and As(III) (<0.00025 µg/g). Conclusion: The data on the arsenic species can be used for future assessments of arsenic exposure and health risks associated with the consumption of Alberta freshwater fish. Funding: NSERC-USRA Email: xsun2@ualberta.ca

Section of Cardiac Surgery, Department of Cardiac Sciences, Cumming School of Medicine, Libin Cardiovascular Institute of Alberta, University of Calgary, Calgary, Alberta, Canada Rationale: Use of extracellular matrix (ECM) biomaterials is an emerging therapeutic strategy to promote cardiac repair and prevent heart failure. We previously demonstrated that implantation of ECM biomaterials on the epicardial surface of infarcted tissue prevents maladaptive remodeling and improves regional functional recovery that extended beyond the passive biomechanical effects of the material. In this study we explored the specific bioinductive mechanism through which ECM therapy enhances myocardial repair following ischemic injury. Methods/Results: ECM-biomaterial was characterized and shown to contain FGF-2 that was released from the ECM upon hydration. To determine the bio-inductive mechanism of the ECMbiomaterial, human cardiac fibroblasts (CF) were isolated from atrial appendage from patients undergoing cardiac surgery. CF are the predominant cell type in the heart that maintain the tissue microenvironment, thereby regulating surrounding cell behaviour and tissue remodeling processes. CF were seeded onto active or biologically inactive (glutaraldehyde-fixed) ECMbiomaterial. Conditioned media was collected and analyzed for angiogenic protein expression using multiplex analysis. CF cultured on active ECM-biomaterial assumed a pro-angiogenic state, showing significantly increased expression of FGF-2, HGF and VEGF. Exogenous FGF-2 inhibition using 1 ď M PD 173074 attenuated the documented pro-angiogenic response, suggesting a paracrine role of FGF-2 as part of the ECM-biomaterialâ&#x20AC;&#x2122;s bioinductive mechanism. Active ECM-biomaterial was utilized in a functional analysis of angiogenesis using human endothelial cells (HUVECs). Here, active ECM-biomaterial was co-cultured with HUVECs embedded in a growth-factor reduced Matrigel. Relative to biomaterial free conditions, active ECM-biomaterial significantly increased endothelial tube formation and branching relative to biomaterial-free conditions. Using a rat model of myocardial infarction (MI), active ECMbiomaterial was applied surgically over the epicardial surface of the infarct and compared against biologically inactive material and sham-treated animals. 14-weeks post-treatment, animals receiving the active-ECM biomaterial showed enhanced neovascularization adjacent to increased levels of vasculogenic growth factors (FGF-2 and VEGF) in the infarcted myocardium relative to the inactive-ECM biomaterial group. Conclusion: These data suggest that an ECM biomaterial applied to the epicardium after ischemic injury can enhance myocardial repair through an FGF-2-dependent mechanism that stimulates vasculogenesis by host cell paracrine effects. Funding: Killam Doctoral Scholarship, Alberta Innovates MD/PhD Studentship

The Yin-Yang of interferon gamma signaling in breast cancer immune surveillance and simultaneous metastatic progression. Stephanie Totten1,2, Ryuhjin Ahn1,2, Josie Ursini-Siegel, PhD1,2 1

Lady Davis Institute, Jewish General Hospital. 2Department of Experimental Medicine, 3 Department of Oncology, Faculty of Medicine, McGill University, Montreal QC, Canada. Rationale: Breast cancer remains the second cause of cancer mortality in women. Responsible for the majority of these deaths, metastasis to secondary organs occurs through a complex cascade of events. Although tumor intrinsic mechanisms dictate whether cancer cells have the potential to metastasize, tumor-stromal interactions must also be permissive for metastasis to occur. Potent anti-tumor immune mechanisms are known to target cancer cells. Yet, unresolved immune responses can yield chronic inflammation that instead promotes metastasis. Along this line, there is evidence for opposing roles for interferon gamma (IFNγ) in cancer as it has been shown to be critical for anti-tumor immune responses but to also contribute to resistance to immunotherapy. Objectives: To elucidate the role of IFNγ in breast cancer progression and lung metastasis. Methods: We used the transgenic mouse MMTV-MT breast cancer model and bred these with IFNγ-/- mice to yield two experimental groups. To address whether IFNγ acts during the later stages of metastasis, MT cell lines were also injected by tail vein into IFNγ-/- and control FVB mice. At humane end-point, lungs were harvested and paraffin embedded. Lung step sections were analyzed for metastases and characterized by immunohistochemistry. Results: While MT/IFNγ-/- mice had an earlier time to primary tumor onset (5.5 weeks) in comparison to MT control mice (7.5 weeks), both groups had comparable primary tumor volumes and tumor bearing days. Yet, only 74% of MT/IFNγ-/- mice developed metastases, whereas 100% of MT mice had metastases. Furthermore, IFNγ-/- had fewer (22 vs. 51, p=0.046) and significantly smaller lung metastases in comparison to controls (0.70mm2 vs 4.4 mm2 p=0.0003). In contrast, when injected by tail vein, IFNγ-/- mice developed markedly more and larger metastases in comparison to controls. Conclusion: We identified a novel role for IFNγ in promoting breast cancer lung metastasis. The reversal of the phenotype in our tail vein experiments strongly supports the hypothesis that IFNγ is acting during the earlier stages of metastasis. This contributes to our understanding of the complex tumor-stromal interactions that are necessary for metastasis and moreover, may allow us to develop improved therapies for patients with metastatic breast cancer.

Funding: CIHR

Email: stephanie.totten@mail.mcgill.ca

Cellular physiology of iron in inflammation and infection processes: novel Fe chelator responses in experimental model systems Karim Wafabc, Nipun Aroraa and Christian Lehmannabcde a

Department of Microbiology and Immunology, bDepartment of Anesthesia, Pain Management and Perioperative Medicine, cDepartment of Pharmacology, dDepartment of Physiology and Biophysics, eFaculty of Computer Science, Dalhousie University. Halifax, Canada. Background: Uveitis is a clinical condition in which the uveal layer of the eye becomes inflamed. If left untreated, uveitis may lead to temporary or permanent visual loss. Currently, corticosteroids are the first line therapy for the treatment of uveitis, however, prolonged use of topical steroids may lead to the development of cataracts and steroid-induced glaucoma. Intracellular iron has been shown to play a role in both acute and chronic inflammation. Iron is required for the generation of oxygen free radicals by polymorphonuclear leukocytes which are responsible for tissue damage during inflammation. Iron chelators are molecules which bind to and sequester iron. By decreasing the levels of iron available to catalyzed oxygen free radical production, we believe that a decrease in tissue damage and monocyte recruitment in states of inflammation is possible. In this study we assess the effects of the iron chelator DIBI on ocular inflammation in an experimental model of non-infectious uveitis. Methods: BALB/c Mice (n=28) were allocated to three different groups receiving intraocular injections of saline/PBS (negative control), LPS/PBS (positive control) or LPS/DIBI (experimental group). Using intravital microscopy, ocular inflammation was assessed by quantifying leukocyte activation, functional capillary density and dysfunctional capillary density. Results: Mice receiving DIBI injections demonstrated a significant decrease in the number of activated leukocytes, improved functional capillary density and decreased dysfunctional capillary density in the ocular vasculature compared to the positive control (LPS/PBS, p<0.05). Conclusion: These results suggest an anti-inflammatory role of the iron chelator DIBI in noninfectious uveitis. Funding: Nova Scotia Health Research Foundation (NSHRF), Chelation Partners Inc., Foundation for Innovation (CFI) and Dalhousie Faculty of Medicine.

Email: kwafa@dal.ca

Common Pathogenic Mechanisms in Age-related Macular Degeneration, Alzheimer’s Disease, Atherosclerosis and Glomerulonephritis Alis Xua, Sijia Caob, Sanjeeva Rajapaksea, and Joanne Matsubarab a

MD Undergraduate Program, bDepartment of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada Rationale: Age-related macular degeneration (AMD) has become the third leading cause of blindness within developed countries and among the elderly, ranked after cataracts and glaucoma. A multitude of risk factors have been recognized amongst AMD patients including aging, smoking and obesity. Due to its complex etiology, current therapeutic approaches have been insufficient at treating AMD. Previous genome-wide association studies of AMD reveal three systems that are closely involved in the pathogenesis of AMD: lipid metabolism, oxidative stress and the inflammatory process. These systems are also involved in Alzheimer’s disease, atherosclerosis and glomerulonephritis. The current literature reports some shared mechanisms in certain aspects of their pathogenesis. A better understanding of the commonalities of these four diseases may provide insight into the unclear etiology and provide novel venues to search for more effective therapies. Objectives: This study attempts to summarize main commonalities in disease pathogenesis of AMD, Alzheimer’s disease, atherosclerosis and glomerulonephritis, establishing a framework of ideas for future research. Methods: Data sources were articles identified through PubMed, Ovid Medline and Google Scholar databases. We summarized the common findings among studies of different diseases and constructed an overall framework of pathogenesis for all four diseases. Results: The current literature supports that AMD, Alzheimer’s disease, atherosclerosis and glomerulonephritis are related in many aspects. Specifically, oxidation processes, lipid deposition, complement activation, and macrophage recruitment are involved in all four diseases shown by genetic, molecular, animal and human studies. Shared risk factors between the diseases further strengthen their connection. Conclusions: An overall framework of pathogenesis can be constructed with the existing evidence. Macroscopic and microscope oxidative stress cause lipid oxidation. Oxidized lipids accumulate at sites of disease development, which activates the complement system and recruits macrophages leading to an increased inflammatory response. As excessive inflammatory damage builds up, pathologic events occur in the eye, brain, blood vessels and kidneys. The commonalities among the mechanisms associated with these seemingly disparate diseases provide novel insight into their future treatment strategies.

Funding: CIHR, Brain Canada

Email: xu.qinyuan@alumni.ubc.ca 66

Sex differences in resistin, chemerin, and chemerin’s receptor in association with carotid atherosclerotic plaque instability Yanofsky R1, Sancho C2, Gasbarrino K1, Jaunet F3, Murray A4, Zheng H1, Lai C5 Veinot J5, Daskalopoulou SS1 Affiliations: 1McGill University, Montreal, Quebec, Canada, 2Université de Montréal, Montreal, Quebec, Canada, 3Polytech Nice-Sophia, Biot, France, 4Western University, London, Ontario, Canada, 5University of Ottawa Heart Institute, Ottawa, Ontario, Canada BACKGROUND Stroke remains a leading cause of death annually, killing more women than men. Despite established sex differences in plaque composition, sex-specific guidelines for carotid endarterectomy (CEA) are lacking. The degree of carotid stenosis, which is the sole indicator for CEA candidacy, is limited in its assessment of plaque stability. Thus, identifying potential sexspecific markers or mechanisms involved in plaque instability is crucial. Herein, we investigated sex differences in the relationship between carotid plaque instability and the expression of proinflammatory adipokines resistin, chemerin, and its receptor, ChemR23. METHOD Clinical information and blood samples were collected preoperatively from subjects undergoing CEA at McGill-affiliated hospitals. Circulating chemerin and resistin were measured using ELISA. Plaque instability was assessed by vascular pathologists using two gold standard histological classifications. Plaques were stained for resistin, chemerin, and ChemR23 using immunohistochemistry. Digital quantification of marker staining was performed using Image-Pro Premier. The percentage of positively stained macrophages/foam cells and their intensity of staining for resistin, chemerin, and ChemR23 was determined using semi-quantitative scales. mRNA expression was assessed by RT-qPCR. RESULTS Circulating resistin was reduced in subjects (n=250) with unstable vs. stable plaques and correlated with stroke in men only (P=0.015). Plaque resistin expression in terms of percent area stained (P=0.026), total area stained (P=0.050), staining intensity on macrophages/foam cells (P=0.002), and percentage of macrophages/foam cells stained (P<0.001) was greater in unstable plaques. However, these associations remained significant in men only (P<0.05). Similarly, greater plaque instability was associated with increased percent area of chemerin staining (P=0.040), and more intense chemerin (P<0.001) and ChemR23 (P=0.013) staining on macrophages/foam-cells, strictly in men. mRNA expression of resistin was significantly increased in unstable plaques (P=0.028), specifically in women (P=0.040). Unstable plaque features were greater in men including increased plaque haemorrhage (P=0.022), lipid core size (P<0.001), plaque inflammation (P=0.007), cap infiltration (P=0.006), and foam cell presence (P<0.05). CONCLUSION We are the first to identify a direct relationship between carotid plaque instability and plaque expression of resistin, chemerin, and ChemR23. Whether a sex-dependant mechanism regulated by these adipokines is implicated in the progression of atherosclerotic plaque instability remains to be examined in future studies.

Funding: Mach-Gaensslen Foundation of Canada, the Sir Edward W. Beatty Memorial Scholarship, the Dr. Clarke K. McLeod Memorial Scholarship 68

Contribution of Pannexin-1 activation to amyloid-beta induced synaptotoxicity and neurotoxicity Albert Yeung1,2, Yufeng Xie1,2, Natalie Lavine1,2, Michael F. Jackson1,2 1

Department of Pharmacology and Therapeutics, University of Manitoba, 2Kleysen Institute for Advanced Medicine, Health Sciences Centre, Winnipeg, MB, Canada Introduction: Calcium dysregulation induced by toxic soluble amyloid-beta oligomers (AβOs) is thought to lead to impairment of synaptic function (synaptotoxicity) and eventual cell death (neurotoxicity) in Alzheimer’s disease (AD). Processes which contribute to the loss of calcium homeostasis can therefore exacerbate the disease process. Pannexin 1 (Panx1) is a large-pore non-selective ion channel broadly enriched in the post-synaptic density of cortical and hippocampal neurons. The open state of Panx1 allows the flux of particles up to 1 kDA, including Ca2+, ATP, and IL-1β. Given its ability to mediate inward Ca2+ currents, we hypothesize that pathological activation of Panx1 contributes to calcium dysregulation in AD. Methods: Naturally secreted AβOs collected from the media of 7PA2 CHO cells expressing the variant of human APP mutation V717F was used as a proven treatment method. Acutely prepared hippocampal slices from Panx1 KO and WT mice were pre-treated with 7PA2 conditioned media (7PA2-CM) and CHO control media (CHO-CM). Changes in long-term potentiation (LTP) between groups was recorded by an established LTP protocol. Primary hippocampal neurons from Panx1 KO and WT mice were also chronically treated with 7PA2CM. Cell death was assessed by LDH assay. Changes in neurite morphology and synaptic markers was assessed via MAP2, synaptophysin, PSD-95 and drebrin immunostaining and Western blot. Results: Panx1 WT slices treated with AβOs showed impairment in LTP, as seen by a significant decrease in fEPSP slope 60 minutes post-tetanus. However, Panx1 KO slices showed no significant change in LTP compared to control treatment. In primary hippocampal cultures treated with 7PA2-CM, dendritic degeneration and cell death were observed by 5 days as seen by a 3-fold increase in LDH release. There was a concomitant decrease in protein levels of the synaptic markers PSD-95 and drebrin. Conclusion: AβOs caused deficits in synaptic plasticity in acute hippocampal slices and induced synapse loss and cell death in primary cultured neurons. Preliminary data suggests that Panx1 contributes to the impairments in LTP. Subsequent experiments will evaluate whether cultured neurons from Panx1 KO mice are protected from AβO treatment. The mechanisms and consequences of Panx1 activation in response to AβOs will also be explored.

Email: yeunga3@myumanitoba.ca

Hyperconnectivity in stem cell-derived neurons of autistic children with SHANK2 haploinsufficiency. Zaslavsky K1,2, Zhang W1, Deneault E1 Zhao M1,2, Ross PJ1, Dedeagac A1, Piekna A1, Wei W 1, Thompson T1, Pasceri P1, Salter MW 1,3, Scherer SW1,2, Ellis J1,2 1

Peter Gilgan Center for Research and Learning, Hospital for Sick Children, Toronto, Canada

Department of Molecular Genetics, University of Toronto, Toronto, Canada

Department of Physiology, University of Toronto, Toronto, Canada

De novo heterozygous deletions and nonsense mutations of SHANK2 (SH3-domain and ankyrin repeats) have been implicated in several autism spectrum disorder (ASD) cases. To investigate their effect on human neuronal function, we generated induced pluripotent stem cells (iPSC) from two children with ASD, one with a SHANK2 nonsense mutation (R841X) and the other a deletion, as well as their parental controls. To isolate the effect of SHANK2, we used CRISPR/Cas9 to generate a homozygous SHANK2 knockout from iPSCs of an unrelated control. We then differentiated these iPSCs into neural precursor cells (NPCs) by inhibition of BMP, Activin/Nodal, and Wnt signaling. The NPCs were then either frozen or further differentiated into cortical neurons. To assess neuronal connectivity in a way that controls for line-to-line heterogeneity, we developed a sparse seeding co-culture assay. Briefly, control and mutant cells are differentially fluorescently labeled and, at four weeks of age, sparsely seeded (2% of total neuronal cell number) into a lawn of unlabeled control neurons on mouse astrocytes. Five weeks after re-seeding, electrophysiological and imaging approaches are used in parallel to assess synaptic connectivity and cell morphology. The frequency of spontaneous excitatory postsynaptic currents was increased in neurons of both autistic children and the SHANK2 knockout, suggesting lowered SHANK2 dosage causes an overabundance of functional synaptic connections. Imaging analyses show that SHANK2 R841X neurons have increased synapse number, synapse density, dendrite length and dendrite complexity. We are currently investigating whether SHANK2 deletion and SHANK2 knockout neurons share the same morphological alterations. Taken together, these data show that SHANK2 haploinsufficiency causes over connectivity in stem cell-derived neurons from children with ASD.

Doxorubicin and Hypoxia Induced Mitochondrial Injury and Necrotic Cell Death of Ventricular Myocytes is Suppressed by Ellagic Acid

Introduction: Polyphenolic compound ellagic acid (EA) found in vegetables and fruits such as pomegranate and blue berry extracts has been reported as having strong antioxidant properties. However, the underlying mechanism by which EA suppresses oxidative injury remains cryptic. In this study, we provide evidence that EA suppressed mitochondrial perturbations and cell death of cardiac myocytes mediated by the chemotherapy drug doxorubicin (Dox) or hypoxia (HPX). Methods: Postnatal rat cardiac fibroblast cells and myocytes were isolated and subjected to primary culture. Cells were treated with Dox, or HPX, in the presence or absence of EA. Control experiments were performed to assess the effect of EA on Dox or HPX on cells. Mitochondrial injury including reactive oxygen species (ROS) production, mitochondrial membrane potential, and mitochondrial morphology was assessed using epiflouroscopy microscopy. Cell viability was determined by immunostaining, using vital dyes to visualize and the number of live and dead cells. At least >200 cells were counted using replicates of n=2-3. Data are expressed as mean ÂąSEM percent dead cells from control. Western Blot analysis was performed for protein expression on cell lysates from cardiac myocytes. The Filters were probed with primary antibodies for Bnip3, DRP1, P-DRP and Îą-actin. Results: Cardiomyocytes treated with Dox or subjected to HPX exhibited a marked increase in the inducible death protein Bcl-2 Nineteen Kilo Dalton Interacting Protein 3 (Bnip3). This coincided with mitochondrial perturbations including increased ROS, loss of mitochondrial membrane potential, increased mitochondrial fission and reduced cell viability compared to vehicle treated control cells. Importantly cardiac myocytes treated with Dox in the presence of EA were indistinguishable from vehicle treated control cells displaying normal mitochondrial morphology, reduced ROS, and preserved mitochondrial membrane potential. This was accompanied by a concomitant reduction in cardiac cell death. Conclusion: The present findings provide the first direct evidence that EA is sufficient to suppress oxidative mitochondrial injury and cell death of cardiac myocytes induced by Dox or hypoxia by a mechanism that impinges upon the death protein Bnip3. By suppressing mitochondrial injury induced, EA may prove to have a beneficial in suppressing cardiac dysfunction in cancer patients undergoing DOX treatment or individuals with ischemic heart disease.

Doxorubicin and Hypoxia Induced Mitochondrial Injury and Necrotic cell Death of Ventricular Myocytes is Suppressed by Ellagic Acid Pegah Afshar* and Lorrie A. Kirshenbaum‡ Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba* and Department of Physiology and Pathophysiology, the Institute of Cardiovascular Sciences, St. Boniface Hospital Albrechtsen Research Centre ‡ Introduction: Polyphenolic compound ellagic acid (EA) found in vegetables and fruits such as pomegranate and blue berry extracts has been reported as having strong antioxidant properties. In this study, we provide evidence that EA suppressed mitochondrial perturbations and cell death of cardiac myocytes mediated by the chemotherapy drug doxorubicin (Dox) or hypoxia (HPX). Methods: Postnatal rat cardiac fibroblast cells and myocytes were isolated and subjected to primary culture. Cells were treated with Dox, or HPX, in the presence or absence of EA. Control experiments were performed to assess the effect of EA on Dox or HPX on cells. Mitochondrial injury including reactive oxygen species (ROS) production, mitochondrial membrane potential, and mitochondrial morphology was assessed using epiflouroscopy microscopy. Cell viability was determined by immunostaining, using vital dyes to visualize and the number of live and dead cells. At least >200 cells were counted using replicates of n=2-3. Data are expressed as mean ±SEM percent dead cells from control. Western Blot analysis was performed for protein expression on cell lysates from cardiac myocytes. The Filters were probed with primary antibodies for Bnip3, DRP1, P-DRP and α-actin. Results: Cardiomyocytes treated with Dox or subjected to HPX exhibited a marked increase in the inducible death protein Bcl-2 Nineteen Kilo Dalton Interacting Protein 3 (Bnip3). This coincided with mitochondrial perturbations including increased ROS, loss of mitochondrial membrane potential, increased mitochondrial fission and reduced cell viability compared to vehicle treated control cells. Importantly cardiac myocytes treated with Dox in the presence of EA were indistinguishable from vehicle treated control cells displaying normal mitochondrial morphology, reduced ROS, and preserved mitochondrial membrane potential. This was accompanied by a concomitant reduction in cardiac cell death. Conclusion: The present findings provide the first direct evidence that EA is sufficient to suppress oxidative mitochondrial injury and cell death of cardiac myocytes induced by Dox or hypoxia by a mechanism that impinges upon the death protein Bnip3. By suppressing mitochondrial injury induced, EA may prove to have a beneficial in suppressing cardiac dysfunction in cancer patients undergoing DOX treatment or individuals with ischemic heart disease.

Funding: University of Manitoba, CIHR and Heart and Stroke Foundation of Canada Email: umafshar@myumanitoba.ca

ORAL PRESENTATIONS P.M. Session

Procurity Lecture Theatre, Apotex Building Main Floor, Room 164 14:15 â&#x20AC;&#x201C; 15:45 Wednesday, June 7th, 2017

Peter Gilgan Center for Research and Learning, Hospital for Sick Children, Toronto, Canada Department of Molecular Genetics, University of Toronto, Toronto, Canada 3 Department of Physiology, University of Toronto, Toronto, Canada 2

De novo heterozygous deletions and nonsense mutations of SHANK2 (SH3-domain and ankyrin repeats) have been implicated in several autism spectrum disorder (ASD) cases. To investigate their effect on human neuronal function, we generated induced pluripotent stem cells (iPSC) from two children with ASD, one with a SHANK2 nonsense mutation (R841X) and the other a deletion, as well as their parental controls. To isolate the effect of SHANK2, we used CRISPR/Cas9 to generate a homozygous SHANK2 knockout from iPSCs of an unrelated control. We then differentiated these iPSCs into neural precursor cells (NPCs) by inhibition of BMP, Activin/Nodal, and Wnt signaling. The NPCs were then either frozen or further differentiated into cortical neurons. To assess neuronal connectivity in a way that controls for line-to-line heterogeneity, we developed a sparse seeding co-culture assay. Briefly, control and mutant cells are differentially fluorescently labeled and, at four weeks of age, sparsely seeded (2% of total neuronal cell number) into a lawn of unlabeled control neurons on mouse astrocytes. Five weeks after re-seeding, electrophysiological and imaging approaches are used in parallel to assess synaptic connectivity and cell morphology. The frequency of spontaneous excitatory postsynaptic currents was increased in neurons of both autistic children and the SHANK2 knockout, suggesting lowered SHANK2 dosage causes an overabundance of functional synaptic connections. Imaging analyses show that SHANK2 R841X neurons have increased synapse number, synapse density, dendrite length and dendrite complexity. We are currently investigating whether SHANK2 deletion and SHANK2 knockout neurons share the same morphological alterations. Taken together, these data show that SHANK2 haploinsufficiency causes overconnectivity in stem cell-derived neurons from children with ASD.

Mutation in Ubiquilin-2 exacerbates ALS/FTD features in a TDP-43 mouse model Vincent Picher-Martel1, Karine Plourde1, Christine Bareil1 and Jean-Pierre Julien1 1

MRI 3-Dimensional Texture Changes in the Motor Cortex Parallel Phenotypic Heterogeneity in Amyotrophic Lateral Sclerosis

Naveet Sohi1, Yee-Hong Yang2, Sanjay Kalra1-4

Rationale: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of the human motor system, with an average survival of 3-5 years from the date of clinical diagnosis. Difficulties in diagnosis, clinical management and drug trials are, in part, a result of extensive clinical heterogeneity in: the site of onset, rate of progression and lack of an objective measures of upper motor neurones. In pursuit of an objective biomarker, many advanced MRI structural modalities have failed to consistently translate ALS phenotypic heterogeneity and disease progression into expected patterns of focal grey matter pathology. Our, recently developed, MRI 3D texture analysis technique has shown significant sensitivity and specificity to structural, and longitudinal, textural change on MRI in ALS. Objectives: Determine the utility of MRI 3D texture analysis as an imaging biomarker to characterize focal changes in the motor strip between patients with limb and bulbar onset, and identify regions that correlate with the degree of clinical disability. Methods: 24 patients with ALS were recruited, scanned and assessed clinically utilizing the ALS Functional Rating Scale-Revised (ALS-FRS-R). MRI 3D texture analysis was utilized to quantify 5 textural values in the primary motor areas and phenotype specific regions of the motor homunculus. Subgroup, correlation and longitudinal analysis was done using SPM8 software. Results: Patients with bulbar onset weakness revealed significant textural differences (p < 0.05, FWE corrected) in the bulbar segment of the motor homunculus compared to the limb segment; conversely, limb onset patients revealed focal textural changes in the limb segment (p < 0.05, FWE corrected). Textural values in the limb segment correlated positively to functional ALSFRS-R scores (p < 0.001), both subgroups revealed diffuse textural changes throughout the motor strip longitudinally (p < 0.001). Conclusions: Our results suggest that 3D MRI texture analysis is a superior biomarker for evaluating phenotypic differences and monitoring progressive changes in the upper motor neurones of ALS patients, when compared to existing structural imaging modalities. The pathophysiological correlation implicated in textural changes remains under study. Funding: David and Beatrice Reidford Research Scholarship, ALS Society of Canada, ALS Association.

Acellular Matrix Biomaterial Promotes Cardiac Repair through an Active Bio-Inductive Mechanism Daniyil A Svystonyuka, Holly E Mewhorta, Jeannine D Turnbulla, Guoqi Tenga, Darrell D Belkea, David G Guzzardia, Daniel S Parka, Sean Kanga, Paul WM FedakaaSection of Cardiac Surgery, Department of Cardiac Sciences, Cumming School of Medicine, Libin Cardiovascular Institute of Alberta, University of Calgary, Calgary, Alberta, Canada Rationale: Use of extracellular matrix (ECM) biomaterials is an emerging therapeutic strategy to promote cardiac repair and prevent heart failure. We previously demonstrated that implantation of ECM biomaterials on the epicardial surface of infarcted tissue prevents maladaptive remodeling and improves regional functional recovery that extended beyond the passive biomechanical effects of the material. In this study we explored the specific bioinductive mechanism through which ECM therapy enhances myocardial repair following ischemic injury. Methods/Results: ECM-biomaterial was characterized and shown to contain FGF-2 that was released from the ECM upon hydration. To determine the bio-inductive mechanism of the ECMbiomaterial, human cardiac fibroblasts (CF) were isolated from atrial appendage from patients undergoing cardiac surgery. CF are the predominant cell type in the heart that maintain the tissue microenvironment, thereby regulating surrounding cell behaviour and tissue remodeling processes. CF were seeded onto active or biologically inactive (glutaraldehyde-fixed) ECMbiomaterial. Conditioned media was collected and analyzed for angiogenic protein expression using multiplex analysis. CF cultured on active ECM-biomaterial assumed a pro-angiogenic state, showing significantly increased expression of FGF-2, HGF and VEGF. Exogenous FGF-2 inhibition using 1 ď M PD 173074 attenuated the documented pro-angiogenic response, suggesting a paracrine role of FGF-2 as part of the ECM-biomaterialâ&#x20AC;&#x2122;s bioinductive mechanism. Active ECM-biomaterial was utilized in a functional analysis of angiogenesis using human endothelial cells (HUVECs). Here, active ECM-biomaterial was co-cultured with HUVECs embedded in a growth-factor reduced Matrigel. Relative to biomaterial free conditions, active ECM-biomaterial significantly increased endothelial tube formation and branching relative to biomaterial-free conditions. Using a rat model of myocardial infarction (MI), active ECMbiomaterial was applied surgically over the epicardial surface of the infarct and compared against biologically inactive material and sham-treated animals. 14-weeks post-treatment, animals receiving the active-ECM biomaterial showed enhanced neovascularization adjacent to increased levels of vasculogenic growth factors (FGF-2 and VEGF) in the infarcted myocardium relative to the inactive-ECM biomaterial group. Conclusion: These data suggest that an ECM biomaterial applied to the epicardium after ischemic injury can enhance myocardial repair through an FGF-2-dependent mechanism that stimulates vasculogenesis by host cell paracrine effects. Funding: Killam Doctoral Scholarship, Alberta Innovates MD/PhD Studentship

The Yin-Yang of interferon gamma signaling in breast cancer immune surveillance and simultaneous metastatic progression. Stephanie Totten1,2, Ryuhjin Ahn1,2, Josie Ursini-Siegel, PhD1,2 1

Lady Davis Institute, Jewish General Hospital. 2Department of Experimental Medicine, 3 Department of Oncology, Faculty of Medicine, McGill University, Montreal QC, Canada.

Rationale: Breast cancer remains the second cause of cancer mortality in women. Responsible for the majority of these deaths, metastasis to secondary organs occurs through a complex cascade of events. Although tumor intrinsic mechanisms dictate whether cancer cells have the potential to metastasize, tumor-stromal interactions must also be permissive for metastasis to occur. Potent anti-tumor immune mechanisms are known to target cancer cells. Yet, unresolved immune responses can yield chronic inflammation that instead promotes metastasis. Along this line, there is evidence for opposing roles for interferon gamma (IFNγ) in cancer as it has been shown to be critical for anti-tumor immune responses but to also contribute to resistance to immunotherapy.

Objectives: To elucidate the role of IFNγ in breast cancer progression and lung metastasis. Methods: We used the transgenic mouse MMTV-MT breast cancer model and bred these with IFNγ-/- mice to yield two experimental groups. To address whether IFNγ acts during the later stages of metastasis, MT cell lines were also injected by tail vein into IFNγ-/- and control FVB mice. At humane end-point, lungs were harvested and paraffin embedded. Lung step sections were analyzed for metastases and characterized by immunohistochemistry. Results: While MT/IFNγ-/- mice had an earlier time to primary tumor onset (5.5 weeks) in comparison to MT control mice (7.5 weeks), both groups had comparable primary tumor volumes and tumor bearing days. Yet, only 74% of MT/IFNγ-/- mice developed metastases, whereas 100% of MT mice had metastases. Furthermore, IFNγ-/- had fewer (22 vs. 51, p=0.046) and significantly smaller lung metastases in comparison to controls (0.70mm2 vs 4.4 mm2 p=0.0003). In contrast, when injected by tail vein, IFNγ-/- mice developed markedly more and larger metastases in comparison to controls. Conclusion: We identified a novel role for IFNγ in promoting breast cancer lung metastasis. The reversal of the phenotype in our tail vein experiments strongly supports the hypothesis that IFNγ is acting during the earlier stages of metastasis. This contributes to our understanding of the complex tumor-stromal interactions that are necessary for metastasis and moreover, may allow us to develop improved therapies for patients with metastatic breast cancer. Funding: CIHR Email: stephanie.totten@mail.mcgill.ca

Common Pathogenic Mechanisms in Age-related Macular Degeneration, Alzheimer’s Disease, Atherosclerosis and Glomerulonephritis Alis Xua, Sijia Caob, Sanjeeva Rajapaksea, and Joanne Matsubarab a

Email: xu.qinyuan@alumni.ubc.ca

Discrepancy between cervical diagnostic biopsy and treatment histology: clinical and viral determinants. Nadège Zanré 1,2, François Coutlée 1,2, Helen Trottier 1,3, Andréanne Pelletier 1,2, Pierre Forest 1, James Bentley 4, Thomas G. Ehlen T5, Laurie Ellit 6, Susie Lau7, Mary Lee5, Marie-Hélène Mayrand 1,2 Centre de recherche du CHM1, Université de Montréal2, CHU Sainte-Justine3, Dalhousie University4, University of British Columbia5, McMaster University6, McGill University7 Rationate: Women who undergo treatment for biopsy-proven cervical precancers may have benign pathology on treatment histology, and as such may have been unnecessarily treated. This may be secondary to spontaneous clearance of precancers, or errors in initial pathology readings. Objectives: To investigate if clinical or virus-related variables are associated with benign pathology after a precancer biopsy. Methods: CoHIPP is a randomized trial conducted in 13 colposcopy centers across Canada, comparing follow-up strategies after cervical precancer treatment. Out of 2162 women recruited at the time of treatment, 394 (18.2%) had benign pathology on LEEP. We designed a case control study that individually matched participants with discrepancy from 10/13 centers (346 cases of discrepancy) with participants with precancer on confirmed on treatment (without discrepancy). We conducted conditional logistic regression to assess the association between routinely collected clinical variables, HighRisk-Human Papillomavirus (HR-HPV) testing (Hybrid Capture 2) and HPV genotyping (Linear Array), with discrepancy. Results: Cases and controls did not differ in terms of age, tobacco use or type of contraception. A negative generic HR-HPV test at baseline was significantly associated with an increased risk of discrepancy (OR: 7.80; 95% CI: 4.88-13.12). The only HPV genotypes associated with a higher risk of discrepancy were types 16, 31, 33 or 52. HPV 16 (compared to no HPV or low risk HPV) had the strongest association against discrepancy (OR: 3.48; 95% CI: 2.43-5.07). Conclusions: Future research should investigate the use of generic HR-HPV testing or genotyping pre-treatment to identity women with precancer biopsy who could be safely followedup instead of being treated. Pathology readings are currently being reviewed. Funding: IRSC, the Terry Fox Foundation Email: nadege.andrea.zanre@umontreal.ca

Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, ON

Institute of Medical Science, Faculty of Medicine, University of Toronto, Toronto, ON

Background: Atherosclerosis is a maladaptive response initiated by the retention of cholesterol-rich low-density lipoproteins, which are rendered pro-inflammatory upon oxidation. While the clearance of oxidized low-density lipoprotein (oxLDL) by macrophages is initially beneficial, excessive accumulation results in cholesterol-laden foam cells that drive disease pathogenesis resulting in the growth of atherosclerotic lesions. The uptake of oxLDL is mediated predominantly by the scavenger receptor CD36 and requires dynamic actin reorganization driven by Rac GTPases. However, external signals that regulate this process remain elusive. The Slit family of secreted proteins, together with their transmembrane receptor Robo, inhibit the migration of a variety of cell types, in part by inactivating Rac and blocking dynamic actin rearrangement. Thus, it is conceivable that Slit-Robo signaling may be harnessed to prevent the progression of atherosclerosis by limiting the uptake of oxLDL and macrophage foam cell formation. Purpose: To investigate the role of Slit2 in atherosclerotic lesion development in ldlr−/− mice and in oxLDL-mediated foam cell formation. Hypothesis: We hypothesize that Slit2 will decrease the size of atherosclerotic lesions and blunt oxLDL-mediated foam cell formation in macrophages by blocking Rac dependent uptake of oxLDL by CD36. Methods: To measure atherogenesis in vivo, ldlr−/− mice were placed on a cholesterol rich diet (CRD) for 6 weeks, then aortas were harvested and mounted en face following perfusion fixation. In parallel experiments, isolated aortas were enzymatically digested and cells analyzed by flow cytometry. Peripheral blood mononuclear cells were isolated by density centrifugation and monocytes polarized for 7 days to macrophages. Foam cell formation was investigated using a neutral lipid probe, BODIPY 497/503, and quantified by immunofluorescence microscopy and flow cytometry. The binding and uptake of oxLDL was investigated using fluorescently (DiI)-labeled oxLDL, and quantified by microscopy and flow cytometry. Results: Exogenous Slit2 decreased atherosclerotic lesion size in ldlr−/− mice placed on a 6 wk CRD. Human and murine macrophages express the Slit2 receptor, Robo-1. Treatment with Slit2 inhibited foam cell formation in human and murine macrophages (P<0.01). Slit2 blocked macrophage binding and uptake of oxLDL (P<0.01) and inhibited oxLDL-mediated activation of Rac1. Conclusions: Slit2 decreases the size of atherosclerotic lesions in ldlr−/− mice and inhibits oxLDL-mediated foam cell formation in both human and murine macrophages. Funding: CIHR MD/PhD Email: ilya.mukovozov@utoronto.ca 83

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Participating Medical Schools

Symposium Contact Information CNMSRS On-Site Contact Info Kimberley Ormiston 204-789-3558 Room 354-3 Chown Building 753 McDermot Avenue Max Rady College of Medicine Rady Faculty of Health Sciences University of Manitoba Winnipeg, MB R3E 0T6 **Email: kim.ormiston@umanitoba.ca

Canad Inns HSC Canad Inns Destination Centre Health Sciences Centre 720 William Avenue Winnipeg, MB Local phone 204-269-9090 Toll-Free 1-888-332-2623 (CANAD)

Sponsoring Agencies

Dr. Paul H.T. Thorlakson Foundation n