RESEARCH & DEVELOPMENT PROJECTS JULY 1, 2001 – JUNE 30, 2004

RESEARCH PROJECTS

Project Title: Activity of ketolides vs. Canadian Gp A Streptococci Co-PI: George Zhanel, Daryl Hoban, Stephen Douthwaite Study description: Study assessing the activity of ketolides vs. Canadian Gp A Streptococci Duration: 2003-2005 Funding: Aventis $30,000

Project Title: Activity of urinary antibiotics against resistant urinary pathogens PI: George Zhanel, Daryl Hoban, Don Low Study description: As above Duration: 2002 Funding: Leo labs $20,000

Project Title: Acute major organ dysfunction in septic patients PI: Bruce Light Study description: As above Duration: 2003 Funding: Glaxo; $85,000

Project Title: Animal models of TSEs PI: S. Booth Collaborators: S.Czub, M. Coulthart, G. Telling, K. Qingzhong Project Description: This project is to establish the infrastructure and expertise within Canada to perform transmission and characterization studies of TSEs in rodent models. Animal models are presently the most sensitive way to establish the presence of TSE infectivity and this capability is essential for the accurate diagnosis and surveillance of TSEs within Canada to inform policy and to assess the risk of transmission between animals and humans. A number of different mouse models, including BSE and CJD that require level III containment, are being established in our laboratory. The study will include primary transmission of Canada’s BSE, CWD and vCJD cases into mice. Transgenic mice containing human, bovine, and elk genes have been previously constructed and reported to show increased sensitivity and short incubation times for transmission. Some of these transgenic models will be used in our studies either in our laboratory or by collaborations with laboratories in the US. Duration: 2003 onwards Funding: Health Canada

Project Title: Antibodies to Burkholderia Type III Secretion System PI: Cangene Corporation (W. Johnson department member) Co-investigators: Defence R&D Canada; Los Alamos National Laboratory – Dr. Paul Jackson Project description: B. mallei and B. pseudomallei , two closely related Gram-negative bacteria, are serious potential bioterrorist agents listed on CDC/MMWR’s category B list. These organisms cause life-threatening infections (Glanders’ disease and Melioidosis, respectively), where antibiotic treatment is sometimes insufficient. The virulence factors of the two species are not well understood, and it is difficult to distinguish these species from each other and also from the closely related avirulent species B. thailandensis. B.(pseudo)mallei is also a civilian problem causing septicemia in Southeast Asia. Another Burkholderia species, B. cepacia, is a more common civilian problem that increases mortality and morbidity of cystic fibrosis patients. To better treat and diagnose B.(pseudo)mallei, we will develop monoclonal antibodies (mAbs) against potential virulence factors: components of the type III secretion system (TTSS). The presence of TTSS has been correlated to virulence of Burkholderia. Also, since several other invasive Gram-negative bacteria utilize TTSS to enter host cells, it is plausible that TTSS are critical for Burkholderia’s virulence as well. TTSS components will be cloned and expressed in E.coli to generate sufficient quantities of purified antigen to generate mAbs. Fully human mAbs will be generated by phage display technology. Initial screening of the mAbs for reactivity will be conducted against a panel of Burkholderia isolates cultivated under different conditions. mAbs of interest will be further evaluated for protective capability in both in vitro and subsequently in vivo small animal models. We will also investigate potential differences of TTSS genes among isolates of Burkholderia. In summary, the primary objectives of the project are: 1) Development of mAbs with utility in diagnosing and treating B.(pseudo)mallei and other Burkholderia species, including B.cepacia; 2)Improved understanding of the pathogenicity of B.(pseudo)mallei as a basis for potential vaccine development. Duration: September 2003 to February 2006 Funding: NIH Grant # 1U10A156383-01; $1,372,000 USD

Project Title: Assay Development and Production team (ADAPT) for the development, validation, production, and distribution of assays for the identification of BT agents. Co-PI: Jody Berry Project Description: Duration: Ongoing Funding: CRTI-DRDC Project; $3,790,000 over 3 years.

Project Title: Assessing the in vitro Herpes simplex virus inhibitory activity of stannous iron. PI: Fred Y. Aoki Collaborators: Dr. Nadia Messiha Project Description: This in vitro project evaluated the effect of stannous iron on the cytopathic effect of Herpes simplex virus. Stannous iron is the purported antiviral component in a topical cream developed by the sponsor for treatment of recurrent cold sore. Duration: 2000-2002 Funding: Embro Research; $8,000

Project Title: Averting multiple organ failure in critically illpatients with sepsis: a pilot study of the effects of glutamine supplementation PIs: Bruce Light, Dr Duerkson, Dr Parry Study description: As above Duration: 2001-2002 Funding: STB Research Foundation; $30,000

Project Title: Bacterial Genomics PI: Michael Mulvey Study description: As above Duration: Ongoing Funding: Canadian Biotechnology Strategy, Health Canada; $650,000

Project Title: Biofilm Build-up in Lumened devices PI: Michelle Alfa Duration: 2001-2003 Funding: Steris Inc $80,000 US

Project Title: Bioinformatics capacity PI : Dr. Lai-King Ng Project description: To enhance the bioinformatics capacity in the National Laboratory for Sexually Transmitted Diseases. Duration: Apr 2001 – Mar 2002 Funding: Office of the Chief Scientist, Health Canada; $15,000 Project Title: Biology and epidemiology of hepatitis C virus PI: J Embree; co-PI: KM Coombs, F Plummer Project description: Hepatitis C virus (HCV) is now recognized as a major disease affecting the blood supply and afflicting 170,000,000 people World-wide. Unfortunately, little is known about basic biology of the virus, how it interacts with cells, and risk factors in its transmission from one infected patient to another person. The goals of this study were to determine factors that influence transmission of HCV in a well-characterized patient cohort. Initially, the prevalence of hepatitis C antibodies in mothers and children as well as in groups of prostitutes, were determined, both with and without underlying HIV infection in order to better understand the factors that contribute to passage of the virus from one person to the next. The strains of virus present in the population were also examined to determine if different strains are present in different populations. Duration: July 2001-June 2002 Funding: Children’s Hospital Research Foundation; $ 22,257

Project Title: Biomedical Proteomics Program: Approaches to the analysis of Disease Progression and Pathogenesis PI: JA Wilkins; co-PI: KM Coombs, W Ens, R Beavis, P Nickerson Project description: Genomics and Proteomics are widely seen as providing powerful analytic methods for determination of alterations in gene expression/utilization. We recently established a Manitoba Centre for Proteomics and have begun to use these mass spectrometric methods for “high-resolution” dissection of virus structure and alterations that take place during assembly and disassembly. We have used this approach to examine details of virus disassembly (Mendez et al., 2003) and to delineate the structure of the RdRp co-factor protein mu2 (Swanson et al, 2002). All other virus proteins are currently under similar analyses. We also are extending these methods to probe protein alterations that take place in cells after and during virus infection. While recent studies are beginning to address genomic alterations (changes in gene expression levels, as measured on “microarrays”), there currently is very little known about what happens to cellular proteins in response to stress. A major new research initiative is to determine changes that take place in viral and cellular proteins during virus infection, both with mammalian reovirus (our model virus) as well as human pathogens like West Nile virus. Duration: October 2001-September 2004 Funding: CIHR; $ 2,025,594

Project Title: Biosynthesis, targeting, and function of hantavirus glycoproteins PI: Heinz Feldman Collaboator: M. Drebot Project Description: Hantaviruses cause two different clinical syndromes; hemorrhagic fever with renal syndrome (HFRS) which is associated with ‘Old World’ (OW) hantaviruses and hantavirus pulmonary syndrome (HPS) which is associated with ‘New World’ (NW) hantaviruses. Hantaviruses are rodent-borne bunyaviruses that are transmitted to humans from chronically infected mice. The worldwide appearance of HFRS and HPS reflects the geographic distribution of the rodent reservoirs of the different hantaviruses. Sin Nombre (SN) virus, the prototype ‘NW‘ hantavirus, is endemic in Canadian deer mice. HPS cases, although rare in Canada, are of considerable public health concern, because of the high mortality rates associated with SN virus infections. Neither a vaccine nor an antiviral treatment is available. Thus, the identification of determinants for pathogenicity would greatly improve our chances to develop urgently needed therapeutic interventions. Transmembrane glycoproteins are important determinants for the pathogenicity of enveloped viruses. In the case of hantaviruses, the medium segment of the tripartite, negative stranded RNA genome encodes the glycoproteins. Studies on hantavirus glycoproteins were hampered in the past by difficulties with virus propagation and the expression of glycoproteins in mammalian cells. To date, knowledge is mainly derived from studies on the glycoproteins of Hantaan (HTN) virus, the prototype OW hantavirus. Preliminary data obtained from infections with Black Creek Canal (BCC) virus, a ‘NW’ hantavirus, has indicated that intracellular targeting and maturation of the glycoproteins may vary between the two groups of hantaviruses. Based on this data, NW hantaviruses may bud from the plasma membrane in contrast to all other Bunyaviridae which bud from Golgi membranes. However, nothing is known about the biosynthesis and structure of NW hantavirus glycoproteins which are important site determinants for the budding of enveloped viruses. Although it seems clear that hantavirus glycoproteins function in receptor binding and fusion, none of the important functional domains have yet been identified for any of the hantaviruses. In this project, the biosynthesis, structure and maturation of the glycoproteins of NW hantaviruses are studied using eukaryotic expression systems. This will allow us to determine if different maturation processes exist between NW and OW hantaviruses. Reverse genetic systems will are used to identify functional domains on the glycoproteins and to define determinants of pathogenicity. In addition, chimeric proteins are generated and used to define the mechanisms of Golgi targeting and retention. Duration: 2002-2004 Funding: CIHR; CIHR Regional partnership program; $149,200

Project Title: The burden of lower extremity complications in persons with diabetes and pre or receiving hemodialysis. PI: John Embil Project Description: As above Duration: 2003-2004 Funding: $6,000

Project Title: Canadian Azole Resistance Study (CARS) Co-PI: George Zhanel and Daryl Hoban Project Description: Eighteen hospital sites representing tertiary care hospitals in Canada are recruited to provide fungemic Candida spp. isolates from blood cultures (1 per patient) plus Candida spp. isolates from the lower respiratory tract and skin and soft tissue infections. Isolates sent to the reference lab (HSC, Microbiology) for antifungal susceptibility testing. Data analysis to include all susceptibility data plus species prevalence data. Duration: Ongoing Funding: Pharmaceutical Industry; $50,000/year

Project Title: Canadian National Fungal Surveillance Co-PI: George Zhanel, Daryl Hoban Study description: Surveillance study assessing antifungal resistance in Canada) Duration: 2001 Funding: Pfizer $120,000

Project Title: Canadian National Fungal Surveillance Co-PI: George Zhanel, Daryl Hoban Study description: Surveillance study assessing antifungal resistance in Canada) Duration: 2002 Funding: Pfizer $120,000

Project Title: Canadian National Fungal Surveillance Co-PI: George Zhanel, Daryl Hoban Study description: Surveillance study assessing antifungal resistance in Canada) Duration: 2003 Funding: Pfizer $120,000

Project Title : Canadian National Fungal Surveillance Collaborators : George Zhanel, Daryl Hoban Study description : A surveillance study assessing antifungal resistance in Canada Duration : 2004 Funding : Pfizer ; $120,000 Project Title: Canadian Network for Public Health Intelligence Co-PI: Amin Kabani Project description: as above Duration: 2002 onwards Funding: CRTI $7,908,234

Project Title: Canadian Network for vaccines and immunotherapeutics of cancer and chronic virus diseases PI: R. Sekaly; U of Manitoba Collaborators: Keith Fowke, Frank Plummer, Jody Berry Study description: The goal of this project is to test HIV vaccines Duration: July 2000 – June 2005 Funding: CANVAC $623,000 .

Study Project: Canadian Nosocomial Surveillance Program Studies PI: Various: Co-Investigators J Embil, J Embree, M Mulvey et al Project descriptions: Nosocomial Cerebral spinal Fluid Shunt-Associated Infections within Acute-Care Institutions: Prospective study of nosocomial CSF shunt infections in participating hospitals across Canada of shunts placed between January 15th 2000 to January 14th, 2002 to determine the incidence, risk factors and outcomes Vancomycin Resistant Enterococci Suveillance in CHEC/CNISP Health Care Facilities: An ongoing surveillance since 1998 of the incidence and changing trends of vancomycin resistance among Enterococci in Canadian sentininel hospitals. MRSA Suveillance in CHEC/CNISP Health Care Facilities: An ongoing surveillance since 1995 of the incidence, changing trends and associated risk factors of MRSA in Canadian sentininel hospitals. Point Prevalence Surveillance for Nosocomial Infections within Selected Canadian Health Care Institutions – February 2002: The variability and cost of nosocomial infections and their prevalence was conducted during a one week period in February 2002 in the 24 CHEC hospitals Duration: Ongoing Funding: Health Canada

Project Title: Canadian Paediatric Surveillance Program – Estimating the Incidence of Severe Combined Iimmunodeficiency (SCID) in Canada (2004-March 2006) PI: Ramsingh R, Project Team: Pelletier L, Probert A, Yacoub W, Junker A, Schultz K, Champagne MA, Long R, Langley J, Embree J Project description: A prospective two year study to estimate the incidence of SCID in Canadian children and, in particular, Aboriginal children in Canada to determine the basic demographics, clinical features and outcomes of children diagnosed with SCID in Canada. Duration: April 2004-March 2006 Funding: Health Canada

Project Title: Canadian Paediatric Surveillance Program – Neonatal herpes simplex virus infection PI: T Wong; Co-Investigators: S Burton, J Embree, R Kropp, M Steben Project description: A study to determine the national incidence of neonatal HSV-1 and HSV-2 infection among Canadian children for the years 2000-2003 and to determine the proportion of those infants with disseminated versus localized disease, the prenatal maternal risk factors among infected infants and to determine the morbidity and mortality attributable to this condition. Duration: October 2000-September 2003. Funding: Health Canada

Project Title: Canadian Respiratory Organism Susceptibility Study (CROSS) Co-PI: Daryl Hoban and George Zhanel Project Description: Twenty-five labs in Canada are recruited to collect, identify and ship to the reference centre (HSC, Microbiology) S. pneumoniae, H. influenzae, M. catarrhalis, S. pyogenes plus all bacteremic S. pneumoniae. At the reference site microbroth MIC testing is performed using carbapenems, ß-lactams, fluoroquinolones plus other comparators, and data analysis conducted. Duration: Ongoing, Currently completing 7th year of study Funding: Pharmaceutical Industry; $400,000/year

Project Title: Canadian Respiratory Organisms Susceptibility Study (CROSS) Co-PI: George Zhanel, Daryl Hoban Study description: Study assessing antibiotic resistance of respiratory pathogens across Canada Duration: 2001 Funding: Multi-industry $550,000

Project Title: Canadian Respiratory Organisms Susceptibility Study (CROSS) Co-PI: George Zhanel, Daryl Hoban Study description: Study assessing antibiotic resistance of respiratory pathogens across Canada Duration: 2002 Funding: Multi-industry $550,000

Project Title: Case control study to examine risk factors for the acquisition of nosocomial ESBLs at CNISP sites Co-Investigator: Michael Mulvey Study description: As above Duration: 2001-2002 Funding: Astra-Zenica; $55,000

Project Title: Characterization of immune mechanisms to resistance to HIV-1 infection PI: Frank Plummer Study description: The overall objectives of our proposed research are to define the mechanisms mediating resistance to HIV-1 and to understand why they develop. Duration: October 2001 – September 2006 Funding: CIHR; $614,000 CAD

Project Title: Characterization of a measles aerosol vaccine device PI: Dr. Allan Coates (Hospital for Sick Children, Toronto) CO-PI: Dr. Graham Tipples Study description: As above Duration: April 2003 – October 2004 Funding: WHO, $20,000

Project Title: Characterization of Mycobacterium tuberculosis Beijing strains in Pakistan using molecular and immunological techniques PI: : Rumina Hasan Co-investigator: Rabia Hussain, Zahra Hasan; Collaborators: Akbar Kanji, Qaisar Hasan, Amin Kabani, Joyce Wolfe, Meenu Sharma Study description: Genotype all the beijing isolates in Pakistan using spoligotyping fingerprinting method. Study the Type1 or Type2 host immune response in patients infected with these strains. Duration: 2003 Funding: AKU intramural grant

Project Title: Characterization of non-Shiga toxigenic Escherichia coli (STEC) isolated from stool in Manitoba during the period 2001 – 2004. PI: Clifford Clark Co-PIs: David Woodward, Paul van Caeseele Collaborators: John Wylie Project Description: The Manitoba Provincial Public Health Laboratory (Cadham Lab) collected non-STEC from diarrheic stools at various locations in Manitoba during the period 2001 to 2004. Initial serogrouping suggested that these isolates might belong to the enteropathogenic Escherichia coli (EPEC) pathotype. Further characterization was undertaken to determine the pathotype and content of signature virulence genes. These data will be combined with the appropriate patient data to assess the epidemiology, the geographical distribution, and public the public health impact of infection with with different non-STEC E. coli pathotypes in Manitoba. The 100 isolates represent atypical EPEC, uropathogenic E. coli (UPEC), enteroaggregative E. coli (EAEC), and diffuse adherent UPEC (DA-UPEC) pathotypes, as well as isolates from which no definitive or signature virulence genes or phenotypes have been detected. Current work is directed toward attempting to determine a denominator value from which the incidence of infection of these organisms can be estimated, further characterization of virulence genes or characteristics, determining whether specific subgroups within each pathotype (eg. among UPEC strains) are clonal, and attempting to assess whether isolates belonging to pathotypes not normally associated with gastrointestinal disease should be considered pathogenic or commensal. Few studies have been undertaken in Canada on E. coli pathotypes other than enterohemorrhagic E. coli O157:H7. The results of this study will provide some insight into non-STEC pathotypes circulating in Manitoba. Duration: 2001 and ongoing Funding status: Funded through the Food Memorandum to Cabinet (Food MC), which provides annual special funding to the Enterics Laboratory for specific or designated projects.

Project Title: Characterization and typing of Listeria monocytogenes in Canada from 1995 – 2003. Co-PIs: Clifford Clark, Jeff Farber (Bureau of Microbial Hazards, Ottawa) Collaborators: Franco Pagotto, Bureau of Microbial Hazards, Ottawa; Paul Sockett, Kathryn Doré, Nadia Ciampa, Centre for Infectious Disease Prevention and Control, Ottawa & Guelph; Louise Ringuette, LSPQ, Québec; John Wylie, Cadham Lab, Manitoba Project Description: All data pertaining to human infection and disease outbreaks caused by Listeria monocytogenes have been collected. All isolates have been or will be tested by serotyping and PFGE with two enzymes. Patient data collected as part of the project includes age, sex, approximate date of collection, and site (eg. tissue, stool). Similarly, summaries of investigations into recent outbreaks of L. monocytogenes in Canada have been prepared. The assembled data will provide 1) an assessment of the utility of typing and subtyping methods for discriminating Canadian isolates, 2) data on the geographic and temporal distribution of different PFGE subtypes, 3) an idea of how often clusters of particular subtypes appear, and 4) baseline data against which changes or trends can be detected. Duration: ongoing Funding: Funded through the Food Memorandum to Cabinet (Food MC), which provides annual special funding to the Enterics Laboratory for specific or designated projects; project proposal was included in funding objectives at time of submission of Food MC.

Project Title: Characterization & validation of a new microbiologic test, the peritoneal fluid bactericidal titre, for use in the treatment of peritoneal dialysis-related peritonitis. PIs: Dr. Sheryl Zelenitsky, Dr. Robert Ariano, Dr. Godfrey Harding Project Description: As above Duration: 2001 Funding: Dr. Paul H. T. Thorlakson Foundation Fund; $17,000

Project Title: Chemokines in immune regulation of human allergic disease PI: Kent HayGlass Co-Investigator: FER Simons Study description: As above Duration: 2004-2008 Funding: CIHR; $665,000 Project Title: Chlamydia immunobiology PI: Xi Yang Study description: As above Duration: 2001-2003 Funding: MHRC; $136,500

Project Title: Chlamydia and gonococcal infection: Immunobiology of the female reproductive tract Co-PI: Robert Brunham, Frank Plummer Study description: The twin goals of the proposed research are to prospectively characterize the mucosal and systemic immune responses that mediate protection against or enhancement of susceptibility to gonococcal and chlamydial infection and disease and to examine the mechanisms by which gonococcal and chlamydial infections alter mucosal immune responses to each other and to HIV. Duration: August 1999 – July 2004 Funding: STD-CRC (University of Washington Subcontract); $624,998 USD

Project Title: Chlamydia trachomatis Polymicrobial infection model PI: Grant McClarty Study description: As above Duration: January 2003 – January 2005 Funding: NIH; $200,000

Project Title: Chlamydia vaccine development PI: Xi Yang Study description: As above Duration: 2003-2007 Funding: NNSF of China; 2003-2005 Y210,000; 2004-2007 Y200,000

Project Title: Ciprofloxacin pharmacodynamics and clinical outcome: The optimal treatment of serious bloodstream infections. PIs: Dr. Sheryl Zelenitsky, Dr. Robert Ariano, Dr. Godfrey Harding Project description: As above Duration: 2003 Funding: Bayer, Inc.; $48,000

Project Title: Clostridium difficle Inactivation Study PI: Michelle Alfa Project Description: Duration: 2002-2003 Funding: St. Boniface Research Centre ($61,000)

Project Title: Clostridium difficle Project PI: Michelle Alfa Project Description: Duration: 2003-2006 Funding: NRC (IRAP) $175,000

Project Title: Community acquired antimicrobial resistant bacteria in northern Canadian communities PI: Michael Mulvey Study description: As above Duration: 2003-2008 Funding: CIHR; $1,457,000 Project Title: Comparative genomics of clinical strains of M. tuberculosis PI: Amin Kabani Project description: As above Duration: 2002 Funding: Office of Biotechnology and Science

Project Title: Comparison of typing and subtyping methods and evaluation of the Oxford Multi-locus Sequence Typing (MLST) and flagellin gene short variable region sequencing (fla-SVR) for Campylobacter surveillance PI: Clifford Clark Collaborators: Louis Bryden, Wilfred Cuff, David Woodward (NML); Bruce Ciebin, Frances Jamieson (Central Public Health Laboratory, Ontario Ministry of Health); Patricia Johnson (Ontario Ministry of Agriculture and Food); Linda Chui (Alberta Provincial Public Health Laboratory); Louise Ringuette (Laboratoire santé Publique de Québec; LSPQ), New Brunswick Emergency Response Centre Project Description: Campylobacter isolates collected as part of the Walkerton outbreak investigation were typed and subtyped at the NML. Epidemiological associations of these isolates to the Walkerton outbreak were known, making this an excellent test system for evaluating the performance of a number of phenotypic and molecular typing, subtyping, and fingerprinting methods. A number of isolates not geographically, temporally, or epidemiologically connected with the Walkerton outbreak were collected to provide a population for comparison of typing and fingerprinting results and to validate conclusions drawn from analysis of outbreak-associated isolates. Duration: Ongoing Funding: Funded through the Food Memorandum to Cabinet (Food MC), which provides annual special funding to the Enterics Laboratory for specific or designated projects; project proposal was included in funding objectives at time of submission of Food MC.

Project Title: The contribution of natural killer cells in the enhances ability of Aboriginals to resolve Hepatitis C virus infection PI: Julia Rempel Co-Investigators: Kent HayGlass and J Gartner Study description: As above Duration: 2003-2006 Funding: CIHR; $195,390

Project Title: Detecting HIV disease progression sooner: the use of IgG subtypes PI: Keith Fowke Study description: As above Duration: January 2004-December 2004 Funding: Manitoba Medical Services Foundation.

Project Title: The determinants and societal impact of the HIV epidemic in India: research program development PI: James Blanchard Co-Applicant: Stephen Moses Study description: This project will develop a research program to generate new research knowledge and methods to improve evidence-based planning of HIV/AIDS prevention and control programs in India. Duration: April 2003-April 2005 Funding: CIHR- GHRI; $99,480 Project Title: Determining pathogenesis of WNV in the North American corvids. PI: Dr. Hana Weingartl Collaborations: The group continues to collaborate with Health Canada, Zoonoses, Dr. Michael Drebot, Dr. Robin Lindsay and other laboratories on establishing and validating serological approaches. Project description: The incursion and establishment of the West Nile virus (Flavivirus, Flaviviridae) on the North American continent became a public and animal health and wildlife issue for Canada. The national and international impact of this technical development work is mainly in the area of human and wildlife health, as from the livestock only horses may develop severe encephalitis. This work is a high visibility issue. Dr. Weingartl lead the establishment of NCFAD testing system for the virus, and reagent development. The project provided novel data on pathogenesis of WNV strain NY99. in the North American corvids, as the index specie fro the WNV surveillance. Development of control samples, reagents and tests for West Nile virus surveillance. Pathogenesis of WNV in North American corvids in the index species used in the WNV surveillance. The work also resulted in number of technology transfers and collaborative projects Duration: April 2001-March 2002 Funding: CFIA TD W0102; $10,000

Project Title: Development of artificial chromosome technology for recombinant antibody protein production in plants. PI: Dr Jim Brandle, AAFC Collaborators: Jody Berry Project description: As above Duration: 2003-2005 Funding: A Matching Industry Initiative, Agriculture Canada/Agrisoma Sponsored Project; $1,249,000

Project Title: Development of a bioinformatics infrastructure for infectious disease profiling PI: S. Booth Collaborators: R.L. Somorjai, C. Bowman W. Cuff and R. Baumgartner Project description: The objective for this project is to establish database resources, both hardware and software, for depositing micoarray images and gene expression data, and to establish a ‘state of the art’ bioinformatics software resource for performing data analysis of microarray images, statistical validation of data and subsequent data mining. Duration: 2002-2005 Funding: Genomics Initiative for Government Laboratories; $540,000

Project Title: Development of diagnostic chips for rapid detection of Category A List pathogens Collaborator: Heinrich Feldmann Project Description: During the recent anthrax attack, thousands of tests were performed to monitor the spread of the disease and prevent further deaths. People found the results confusing, and were not always comforted by them. A rapid and reliable method to diagnose infection by all ‘Category A List’ agents is now needed. This method should also allow one to diagnose infections by pathogens that produce similar symptoms so that they have to be considered in the differential diagnosis of A list agents. This is required to respond to a bioterrorism attack, a naturally occurring disease outbreak, or the importation of single cases of disease. All of the ‘Category A List’ agents cause acute diseases with unreliable host immune responses during the viremic/bacteremic, infectious stages of the illnesses. Thus, pathogen detection assays are the primary choice for rapid diagnosis. In principle, pathogens can be detected with antibodies (e.g., antigen detection ELISA), which have to be made and characterized. This can be an expensive and time-consuming process. The alternative is to develop methods for detecting genomic DNA or RNA, or transcripts derived from the infectious agents of interest. This strategy is potentially faster to implement. Genomic sequence is available for all of the agents on the ‘Category A List’. This information has already been used to develop PCR-based assays. Typically, PCRs for different pathogens have to be run separately from one another due to differences in amplification conditions. This makes it difficult to assay a large number of samples for many pathogenic species. Furthermore, the PCR methods that are currently in use are plagued by false-positive results due to carryover contamination, and are limited in their ability to detect rare transcripts. The latter problem has made it hard to multiplex the PCR reactions. The more templates you try to amplify, the more difficult it becomes to achieve optimal reaction conditions for each individual pair of primers. Nonspecific interference of primers with one another and reduced specificity add to the problem. We propose to use DNA microarray technology for rapid detection of ‘Category A List’ pathogens and for additional agents as well. We feel that we can achieve the sensitivity and specificity needed to make the method robust enough so that it Duration: 2003-2005 Funding: National Institutes of Health (NIH) ; US $ 160,000

Project Title: Development and implementation of a molecular pathotyping of Newcastle disease virus and avian influenza virus as diagnostic methods. PI: Hana Weingartl Co-PI: Dr. John Pasick Study description: Molecular pathotyping of Newcastle disease virus(NDV) and avian influenza(AIV) isolates is an essential tool in predicting the pathotype of the virus. It will immediately identify pathogenic strains of NDV and AIV even though they may not appear as such at the original isolation. It is absolutely critical for CFIA to have such a test in place, also as it is now requested by O.E.I. to supply sequence data with NDV and AIV H5 and H7 outbreak reports. Protocols were developed for molecular pathotyping of Newcastle disease virus and AIV, subtypes H5 and H7;phylogenetic analysis was introduced into the diagnostic repertoire as a tool to assist with the analysis of sequencing data. Number of novel sequences of the H5 and H7 subtype of AIV and of the NDV fusion protein were determined. This work was expanded into phylogenetic analysis of cormorant NDV isolated in Canada in years 1990 - 2000, which could help better understanding of NDV in Canadian wild bird population. Two publications resulted from this work. Impact on the clients is prevention of incursion of foreign animal disease into Canada, and prevention of an epidemic outbreak. Duration: April 1999 – March 2002 Funding: CFIA TD W9802; $30,000

Project Title: Development of Novel IL-4 and IL-13 peptide based vaccines for the prevention and treatment of allergic diseases PI: Z Peng Co-Investigator: Kent HayGlass Study Description: As above Duration: 2003 - 2005 Funding: MICH, $10,000; $50,000

Project Title: Development of pluripotent P19 EC cells as an in vitro model to investigate the mechanism of prion provoked neurotoxicity. PI: J. David Knox Collaborators: Margot Plews, Sharon Simon, Jillian McMaistre, J. David Knox (PI). Study description: Transmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative disorders of humans-Creutzfeldt-Jacob disease (CJD) and animals- sheep scrapie, chronic wasting disease (CWD) and bovine spongiform encephalopathy (BSE). Thought to be the main infectious agent, PrPres, is a misfolded isoform of a normal protein, PrP or prion protein found in the brain and other organs of the host. The relationship between PrPres and neurodegeneration has been established, yet the mechainism is unclear. A considerable amount of data linking PrP to oxidative stress has been accumulated 1. I propose to study the mechanisms involved in the neurotoxic effect of prion proteins using an in-vitro model. My hypothesis is that PrPres causes nerve cell death by increasing oxidative stress. P19 cells are pluripotent stem cells, which under retnoic acid (RA) induction differentiate into neuronal, glial, and non-neuronal cells 2 The composition of the culture will be confirmed by real-time PCR looking for the cell-type specific transcripts. I will then clone the green fluorescent protein (GFP) gene into a plasmid, under regulation of the neuron specific promoter T1 tubulin and transfect the construct into the P19 cells 3. When differentiated with RA the neuronal P19 EC cells will express GFP. Fluorescent cells can be counted and isolated by fluorescent-activated cell sorting (FACS). The established cultures and neuronal tracking method will enable the effect of increasing PrPres concentrations on neuronal cell death to be determined. The PrPres will be isolated from the brains of mice infected with the scrapie strain ME7. Uninfected brains will be used as a control. Following exposure of the neuronal P19 cells to toxic amounts of PrPres I will use real-time PCR and biochemical assays to search for markers of oxidative stress 4. Oxidative stress results from an imbalance in cellular anti-oxidant/oxidant homeostasis . Under oxidative stress the cellular content of reactive oxygen species (ROS) greatly increases . Glutathione (GSH) is the main anti-oxidant of the brain. To investigate the role of oxidative stress in PrPres toxicity I will investigate the modulating effects of two drugs, Buthionine sulfoximine(BSO) and geranylgeranylacetone (GGA). BSO is an inhibitor of the rate limiting enzyme in GSH synthesis, while GGA, through a translational mechanism, results in increased glutathione production 5. The hypothesis predicts that the addition of BSO will sensitize neurons to PrPres provoked cell death and that the addition of GGA protects cells from PrPres provoked cell death. Glutathione levels will be measured by a modified spectrophotometric technique5. Recently it has been proposed that TSE diseases are part of a larger family of neurodegenerative diseases including Alzheimer’s disease.This is based on histological similarities that include the disease associated accumulation of misfolded proteins in the brain and biochemical evidence of oxidative stress. The convergence of these diseases at the point of neuronal loss indicates that an understanding of the mechanism of neuronal loss in TSE diseases may be more widely applicable to neurodegenrative diseases in general. This will contribute to the health of an aging Canadian population. Duration: Started 2004

Project Title: Development of a Rapid Method for the Genotyping of Human Papillomavirus (HPV) Using the Luminex xMAP Technology PI: Alberto Severini Collaborator: Magdi Dawood Study description: Development of a multiplex method for typing the over 40 types of genital human papillomaviruses. The Luminex method is much faster than currently available hybridisation methods and it is amenable to automation. With the advent of HPV DNA testing along side cervical cytology and with the very promising clinical trials of HPV vaccines, high throughput typing of HPV isolates will soon become a pressing need. Duration: 2003-2004 Funding: Health Canada Office of the Chief Scientist; $33,412.50

Project Title: Development of rapid vaccination strategies in large animals. PI: Jody Berry Project Description: Duration: 2004 Funding: CFIA Quickstart program; $4000

Project Title: Development of a real-time Web-based surveillance (WEBS) for CNISP surveillance program Co-Investigator: Michael Mulvey Study description: As above Duration: 2002 Funding: Pharmacia-Upjohn; $150,000

Project Title: The Development of Recombinant Monoclonal Antibodies (MAbs) for Treatment and Detection of BioTerrorism Agents [BTA] CO-PI: Jody Berry; Amin Kabani, Keith Fowke Project Description: As above Duration: 2003-2006 Funding: CRTIDRDC Project; $5,730,640 Project Title: Development of therapeutic antibodies to neutralize the virulence determinants of the sexually transmitted pathogen Chlamydia trachomatis. PI: Grant McClarty Collaborators: Jody Berry Project Description: As above Duration: 2002-2003 Funding: Office of the Chief Scientist, Health Canada.

Project Title: Development of therapeutic vaccine for chronic hepatitis B PI: Jingxin Cao Project Description: The capability of recombinant vaccinia viruses to effectively prime and stimulate CTL has made them extremely useful vector for recombinant vaccine studies. Therefore, this proposal is to develop a recombinant vaccinia virus based therapeutic HBV vaccine that expresses a fusion protein containing mycobacterial HSP 70 and HBV proteins. Duration: 2000 onwards Funding: Health Canada internal funding; approximately $10,000 per year.

Project Title: Development of a virulence gene microarray for enteric pathogens PI: Clifford Clark Collaborators: Gehua Wang, MD, Louis Bryden, M.Sc., NLEP, Health Canada; Kris Rahn, M.Sc., Susan Read, DVM, Ph.D., Roger Johnson, DVM, Ph.D., Cornelius Poppe, DVM, Ph.D., Kim Ziebell, M.Sc., Laboratory for Foodborne Zoonoses (LFZ), Health Canada; Victor Gannon, DVM, Ph.D., LFZ, Health Canada; John Nash, Ph.D., Institute for Biodiagnostics, NRC; Sabah Bidawid, Ph.D., Jeff Farber, Ph.D., Bureau of Microbial Hazards (BMH), Health Canada; Roland Brousseau, Ph.D., National Research Council, Montreal; Josée Harel, Ph.D., John Fairbrother, Ph.D., University of Montreal Project description: The objective of this project is to develop a DNA microarray carrying probes for known strain-specific genes and virulence genes of bacterial and viral food- and water-borne pathogens, including those on mobile genetic elements or those that may not be present in DNA microarrays prepared using information from sequenced genomes. This microarray has potential as a diagnostic and monitoring tool. Data gathered from use of this microarray will contribute to an understanding of the role(s) of specific virulence factors in pathogenesis of the organism and will aid in the detection of co-ordinately regulated groups of virulence genes that have defined roles in infection. The array will facilitate detection of strain-specific genes and virulence genes that do not normally occur in given pathotypes, contributing to the identification of emerging pathotypes with enhanced virulence. Duration: 2002-2005 Funding: Health Canada Genomics/Biotechnology Fund; $300,000

Project Title: The diagnosis of congenital Cytomegalovirus infection by PCR of newborn dried blood spot specimens PI: Dr. Marilyn Kirkpatrick, Pathology Resident Co-PI: Doug Milley; Supervisor: Paul VanCaeseele; Collaborator: Robert Thompson Study Description: Cytomegalovirus (CMV) is the most common congenital viral infection, with observed infection rates varying between 0.2 to 2.5% of all newborn infants. Ninety percent of congenitally infected newborns are asymptomatic, and at least 10 to 15% of these newborns are at risk for subsequent abnormalities including sensorineural hearing loss. Congenital CMV infection is currently diagnosed by detecting virus in urine and other body fluids during the first three weeks of life. Positive culture after three weeks of age may be due to either congenital or perinatally acquired infection. Recently, PCR of CMV DNA extracted from newborn dried blood spot (DBS) specimens has been used to diagnose congenital infection, although the literature notes that further studies are required to evaluate the sensitivity and specificity of the methods. Our goal is to do a feasibility study on a method of performing PCR on CMV DNA extracted from stored dried blood spots at our centre, Cadham Provincial Laboratory (CPL). The method is a modification of an existing validated diagnostic in-house method of CMV PCR that has been used since January 1998 for CMV detection in Manitobans. This procedure is valuable because it would allow a retrospective diagnosis of congenital CMV to be made when suggestive symptoms (eg. sensorineural hearing loss) are present. As well, a diagnosis of congenital CMV in an asymptomatic patient would allow physicians and parents to discuss prognostic issues, and possibly increase surveillance for adverse sequelae. Once the method has been validated, it may permit us to redefine congenital CMV infection based on an adequate numbers of specimens for statistical evaluation. Finally, this procedure could provide useful epidemiologic data regarding rates of congenital CMV infection in Manitoba. Duration: June 2002 – December 2003 Funding: Cadham Public Health Laboratory - Internal Funding

Project Title: Direct detection and identification of bioweapon nucleic acids based on cationic polymers Co-investigator: Michael Mulvey Study description: As above Duration: 2003-2007 Funding: CRTI $369,362

Project Title: DNA chip technology for enhanced surveillance of human pathogens: genotyping and identification of antimicrobial resistance genes CoPI: Michael Drebot; Lai-King Ng Project description: To develop a method using DNA microarray for detecting virulence genes expression of enteric pathogens from clinical stool specimens. Duration: Apr 1999 – Mar 2002 Funding: Office of the Chief Scientist, Health Canada; $140,000; Health Canada Genomics R&D Biotechnology Grants; $150,583/year

Project Title: DNA microarrays for studying epidemic MRSA PI: Michael Mulvey Study description: As above Duration: 2001 Funding: Office of the Chief Scientist; $53,000

Project Title: Double-blind, placebo-controlled trial of long term valaciclovir in patients with Herpes simplex encephalitis. PI: Dr. Richard Whitley, Birmingham, Albama Collaborators: Fred Aoki is one of many collaborators Project Description: This is a multiple-centre study evaluating the effect of three months of valaciclovir administered after completion of a conventional 21-day course of intravenous acyclovir for treatment of Herpes simplex encephalitis, on the long term neurological outcome of the patient. Duration: 1999 onwards Funding: NIAID, USA, $15,000 US

Project Title: Drotrecogin alpha activated in adult patients with early stage sepsis PI: Bruce Light Study description: As above Duration: 2003 Funding: Lilly Pharmaceuticals; $23,000

Project Title: Edible vaccines PI: Anton Andonov Project Description Duration: Ongoing Funding: Public Health Agency of Canada Project Title: The effect of infection on allergy PI: Xi Yang Study description: As above Duration: 2001-2004 Funding: CIHR; $371,281

Project Title: Effects of antibiotics on selection of resistant flora PIs: George Zhanel, Daryl Hoban, Dr G, Garber (Ottawa) Study description: Study assessing the impact of antibiotics in vivo on colonic flora Duration: 2003-2005 Funding: Aventis; $680,000

Project Title: Epidemiologic Projections of Diabetes and its Complications: A Population Based Study. Investigators: Drs. J. Blanchard, P Orr, S. Ludwig, L. Nicolle. Study description: As above Duration: January 2000 - ongoing Funding: Sellers Foundation, $30,000,

Project Title: Epidemiology of community acquired methicillin-resistant Staphylococcus aureus (group Grant) PI: Dr Marie Louie Collaboratosr: John Embil Project description: As above Duration: 2001-2002 Funding: CIHR; $250,000

Project Title: The epidemiology, health care utilization, and cost of inpatient and outpatient care by patients with diabetes and lower extremity complications. PI: John Embil Project description: As above Duration and Funding: 2001-ongoing; 2001- 2002 Health Science Centre - $24,985; 2002-onward Eli Lillie - $50,000 x 1 year then $25,000/year

Project Title: Epidemiology of Herpes zoster, its treatment and its complication, post-herpetic neuralgia in Manitobans: A population-based study using Manitoba Medicare Claims Data and Pharmacare data. PI: Fred Y. Aoki Collaborators: Dr. Charlyn Black Project Description: Initiated in 1999 during my sabbatical year, this project is continuing to describe the way in which Manitoba physicians treat Herpes zoster with antiviral drugs and the effect that has on chronic analgesic use as a surrogate marker for post-herpetic neuralgia, using administrative data bases. Duration: Ongoing since 1999 Funding: Clinical Studies Fund; Aoki Research General Research Funds

Project Title: Epidemiological study of external ventricular drain (EVD) central nervous system (CNS) infections at a Canadian tertiary care centre PI: John Embil Project description: As above Duration: 2001-2002 Funding: Health Sciences Hospital Foundation; $15,000 Project Title: Establishment of a Centre for Systems Biology PI: JA Wilkins; co-PI: KM Coombs, W Ens, R Beavis, P Nickerson Project description: This award, from the Canadian Foundation for Innovation, is being used to equip ~7,000ft2 of lab space on the 7th floor of the John Buhler Research Centre for a shared lab facility (principally JAW and KMC) that will combine cell culture, virus culture, immuno-biology and monoclonal antibody production and usage, and new state-of-the-art mass spectrometric analyses of biomedical problems (to complement funded research project detailed previously). Duration: May 2004 onwards Funding: CFI; $ 3,043,630

Project Title: Establishment of a Pox and Hepatitis C Virus (HCV) Chimera System to Study HCV Replication in Tissue Culture and Pathogenesis in a Mouse Model PI: Jingxin CAO Project Description: To establish a pox and HCV chimera system based on a replication defective (in mammalian cell) vaccinia virus MVA (Modified Vaccinia Ankara). The major objective of establishing such a system is to recapitulate the replication events of HCV in both tissue culture and a small animal model, such as mice. Such a system will be extremely valuable in treating HCV infection and developing preventive/therapeutic vaccines. Duration: 2001-2002 Funding: Office of Chief Scientist, Health Canada; $36,000

Project Title: An ethnographic assessment of female sex workers in rural India PI: James Blanchard Co-Investigator: Stephen Moses Study description: The broad objective of this project is to gather information that will support the development of culturally appropriate and effective HIV prevention interventions among rural female sex workers in northern Karnataka State. Duration: January 2000-August 2001 Funding: World AIDS Foundation; $90,000 USD

Project Title: Evaluation of GM-CSF for acute radiation syndrome PI: Cangene Corporation (Wendy Johnson, department member) Collaborators: Health Canada Radiation Protection Branch, University of Maryland Project description: Radiation overexposure is considered a potential threat to both civilian and military personnel in various circumstances. Radiation exposure has been reported in a variety of accidents that include accidental X-ray exposure and nuclear plant accidents, as well as a military training accident reported in 2000. Deliberate exposure in a military or terrorist situation must also be considered, due to the proliferation of global nuclear capacity and traffic in spent nuclear fuels. In humans exposed to radiation, the use of cytokine therapy (therapy using regulating proteins) has not been systematically evaluated. However, laboratory, animal and clinical studies have suggested a role for hematopoietic stem cell modulators (cytokines that regulate the development of blood cells). In particular, animal studies have demonstrated that GM-CSF (granulocyte-macrophage colony-stimulating factor, which is known to stimulate the production of white blood cells) is useful in mitigating the effects of sub- lethal radiation exposure where some viable early stem cells remain. All currently available cytokines have proved ineffective where the dose of radiation is such that all stem cells are eliminated. Cangene Corporation is developing a recombinant human (rh) form of GM-CSF, LEUCOTROPIN™, and will shortly submit this drug for licensure in Canada. The goal of the current project is to demonstrate the utility of LEUCOTROPIN™in restoring the body's ability to produce white blood cells following radiation-induced damage to the bone marrow. The primary application for GM-CSF in radiation exposure would be for the early treatment of patients exposed to low- to medium-dose radiation. However, the drug may also be useful for protection of individuals likely to be exposed to radiation, such as rescue workers.To achieve the these goals, Cangene will produce LEUCOTROPIN™according to its established cGMP process at 2,100 L scale in the company's manufacturing facility. The material will be evaluated in the pilot and full animal studies. A parallel study to evaluate a more stable PEGylated GM-CSF developed by Cangene for single-dose administration will also be conducted. The pilot study will be conducted in cynomolgus monkeys to determine an effective low-to-medium and medium-to-high single-dose radiation cycle to compromise immune system. Confirmation of drug dose will be conducted as well. The objective of the full animal study would be to show efficacy of GM-CSF in the early treatment of low-to-medium dose radiation exposure. Monkeys will be subjected to a series of increasing levels of single exposure to radiation with recovery cycles. The placebo-controlled study will evaluate the effect of GM-CSF on relief of neutropenia. The primary end point will be a surrogate marker for immune system competence while secondary end points will include reduction in infections and transfusion requirements. Based on the efficacy results, Cangene will file a supplemental New Drug Submission for approval to add the indication of treatment for low-to-medium dose radiation exposure to the LEUCOTROPIN™file. Duration: September 2003-August 2005 Funding: $1,200,000

Project Title: An evaluation of local strains of Blastomyces dermatitidis PI: John Embil Project Description: As above Duration: 2002-2003 Funding: Manitoba Medical Services Foundation; $12,500

Project Title: Evaluation of measles and rubella serological assays PI: Dr. Graham Tipples; Study description: As above Duration: February – October, 2004 Funding: Health Canada and WHO and PAHO in-kind contributions; $50,000.

Project Title: Evauation of modified vaccinia virus Ankara based recombinant SARS vaccine in ferrets PI: Jingxin Cao. Project Description: Severe acute respiratory syndrome (SARS) caused by a newly identified coronavirus (SARS- CoV) remains a threat to cause epidemics as evidenced by recent sporadic cases in China. In this project, we evaluated the efficacy and safety of two SARS vaccine candidates based recombinant modified vaccinia Ankara (MVA) expressing SARS-CoV spike and nucleocapsid proteins in ferrets. No clinical signs were observed in all the ferrets challenged with SARS-CoV. On the other hand, vaccination did not prevent SARS-CoV infection in ferrets. In contrast, immunized ferrets (particularly those immunized with rMVA-spike) exhibited significantly stronger inflammatory responses and focal necrosis in liver tissue after SARS-CoV challenge than control animals. Thus, our data suggest that enhanced hepatitis is linked to vaccination with rMVA expressing SARS-CoV antigens. Duration: 2003-2005 Funding: Health Canada SARS funding

Project Title: Famciclovir one-day treatment for recurrent genital herpes. PI: Fred Y. Aoki, MD Co-Investigators: Greg Hammond and Pam Orr Project description: Double-blind, placebo-controlled trial comparing conventional 5-day treatment of genital herpes with famciclovir to an experimental regimen comprising 1000 mg BID for one day only. Duration: May-September 2004 Funding: Novartis Pharmaceuticals- Novartis FAM810A study: $57,750

Project Title: FluInsureR protective efficacy. PI: Fred Y. Aoki, MD Co-Investigators: Greg Hammond and Pam Orr Project description: Placebo-controlled, double-blind trial of the protective efficacy of intranasal inactivated proteosome adsorbed polyvalent influenza vaccine for prevention of clinical influenza. Duration: Winter 2003-2004 Funding: IDB Pharmaceuticals; $64,680 Project Title: A Fluorescence Imaging System and IR spectrometer for developing further applications of fluorescence and bioluminescence in the diagnosis of infections in live animals. PI: Tim Booth Co-Investigator: Mike Jackson Study Description: Sampling for infectious disease can be a problem, for example spinal fluid has to be tapped for viral encephalitis tests. This is a difficult and potentially dangerous procedure for the patient. Development of devices that rely on biophysical principles, and can diagnose non-invasively, avoids having to take tissue fluids or biopsies and also obviates the need for traditional biochemical reagents that are difficult to standardise. Herpes simplex virus type 1 (HSV-1) is the most common cause of focal encephalitis in humans. The death rate from this disease is high, even with antiviral therapy. We have used a well-established mouse model of Herpes simplex encephalitis (HSVE) to obtain preliminary information on conditions for detection of HSV proteins in mouse brain tissue. Duration: 04/2002 - 04/2003 Funding: Strategic science fund, Office of the Chief Scientist, Health Canada, $28,135.35

Project Title: Fluoroquinolone-resistant E.coli Collaborators: George Zhanel, Daryl Hoban, Dr. J Johnson (Chicago); Lindsay Nicolle, Godfrey Hading, Dr Don Low (Toronto) Study description: A North American surveillance study assessing the prevalence of fluoroquinolone-resistant E.coli Duration: 2003-2005 Funding: Procter Gamble; $350,000

Project Title: A Genetic “Knock-Down” Approach to Identify Host Cellular Factors Essential for Infectious Agent Replication and Pathogenesis Co-PI’s M. Carpenter, M. Czub and D. Knox Project Description: This project studies the application of a functional genomics screen (inverse genomics) to identify host cellular gene products essential for infectious agent infection, replication and pathogenesis in cell culture. Inverse genomics allows exploitation of genetic knock-outs for the analysis of complex mammalian systems. Our research initiative is most interested in targets with the potential to become therapeutic factors. Thus, the inverse genomics approach offers a further layer of selection in that host factors, which are targeted by the ribozyme and which are essential for the host, are rapidly screened out of the selection pool. Inverse genomics has previously been used to identify genes involved in cancer cell migration and metastases, in TNF- alpha mediated apoptosis, and in the identification of host genes that regulate hepatitis C virus translation. Duration: 2003-2004 Funding: Office of the Chief Scientist (Health Canada); $36,952

Project Title: Genetic and phenotypic characterization of resistance to HIV-1 infection PI: Frank Plummer Study description: This project arises out of our work on characterizing mechanisms of resistance to HIV-1 among a cohort of highly exposed prostitutes in Nairobi. In this project, we will conduct an epidemiologic study to determine the frequency of the HIV-1 resistance phenotypes in the families of HIV-1 resistant women and identify potential modes of inheritance of the phenotype. Duration: October 2001 – September 2006 Funding: CIHR; $587,982 CAD

Project Title: Genetic Susceptibility to HIV Infection PI: Keith Fowke Study description: As above Duration: July 2002-June 2003 Funding: Thorlakson Foundation. $23,740 Project Title: Genomic approach to detect and differentiate verotoxigenic Escherichia coli PI : Dr. Lai-King Ng Project description: To use a genomic approach to identify genes and loci suitable for detection and typing of verotoxigenic E. coli, VTEC for “real-time” surveillance. The typing approach to be used is a multilocus sequence typing system, MLST. MLST will be easier to compare inter-laboratory data with less laborious analysis and will provide a higher discriminatory power than current methodologies. Duration: Apr 2003 – Mar 2005 Funding: Office of Biotechnology and Science, Health Canada; $97,250

Project Title: Genomics of simian herpes simplex viruses PI: Alberto Severini Study description: Comparative sequencing of primate simplexviruses and study of gene expression by DNA microarrays, in order to elucidate the exact evolutionary history of these viruses and identify pathogenetic mechanisms and candidate virulence genes. Duration: March 2003 – March 2005 Funding: Health Canada Genomic Research and Development Fund; $97,000

Project Title: Genomics of Infection and Immunity: PI: Frank Plummer Sub-Project title: Host Genomics of Pathogen Infection Interactions HIV1, HCV, Prions Co-PI’s, Michael Carpenter, J. Cao and David Knox. Project Description: A multi-collaborative grant which seeks to: 1) Use DNA microarray technology to characterize differences in gene expression between host “infected” and “uninfected” samples for HIV, HCV and Prion associated pathologies, (2) Use single nucleotide polymorphism (SNP) chip technology to identify host elements or markers associated with differential sensitivity to pathogen infection (HIV-1), (3) Compare microarray expression data for different pathogens to identify common host immune/cell signaling responses to infection by HIV-1, HCV and Prions, (4) Identify bio-markers associated with increased/decreased pathologies that may be developed for future diagnostic testing as well as provide potential therapeutic targets (HCV, Prions). This work also utilizes high-resolution proteomics techniques to study changes in cellular proteomes. Duration: 2002-2005 Funding: Office of Biotechnology, Health Canada; 233,000/year Sub-project title: The Use of Microarrays for Comparative Genomics and Expression Profiling to Study Virulence in Clinically Significant Canadian Strains of Bacterial Pathogens PI : Dr. Michael Mulvey Co-PI: Dr. Lai-King Ng Project description: Despite great progress in medicine during the past 100 years, human infectious diseases remain a significant cause of illness and approximately 30% of deaths world-wide. In this proposal, we have selected four bacterial disease pathogens in which Health Canada has been monitoring for decades. The global impact on human health for these pathogens is briefly detailed below. Mycobacterium tuberculosis (MTB) is considered by the World Health Organization (WHO) to be the number one infectious killer in the world, causing three million deaths annually. In Canada, the number of MTB cases in the aboriginal population is 20 times the non-aboriginal rate, higher than those seen in some parts of Africa and Asia. Chlamidia trachomatis is the world’s leading cause of preventable blindness, affecting 400 million people, and also the most common sexually transmitted bacterial pathogen, with an estimated 90 million cases occurring each year world-wide. Neisseria gonorrhoeae is the causative agent for gonorrhea, pelvic inflammatory disease (PID), tubal infertility and ectopic pregnancies. The costs associated with female gonococcal (GC) infections are estimated to be in excess of $43 million annually. According to the WHO, hospital acquired “superbugs” kill about 14,000 patients a year in the U.S. alone with methicillin-resistant Staphylococcus aureus (MRSA) the most prevalent. Canada has also observed a dramatic increase in MRSA within our hospitals with rates increasing eight-fold since 1995. The cost associated with treating these patients in Canadian hospitals in 1997 has been estimated to be approximately $60 million but this number could exponentially increase as the rates continue to rise. The elucidation of the complete genome sequence of the above mentioned pathogens has enabled the production of DNA microarrays, or biochips, which are at the cutting edge of new technologies related to the study of gene expression and comparative genomics. Instead of detecting and studying one gene at a time, microarrays allow thousands or tens of thousands of specific DNA or RNA sequences to be examined simultaneously on a small glass or silica slide only 1-2 cm square. Although the principles of specific DNA and RNA detection remain unchanged, the greatly increased scale on which this can be achieved with DNA microarrays makes it possible to tackle qualitatively different questions. The past five years have seen a dramatic rise in the use of DNA microarrays for biomedical research, in some cases with immediate applicability to clinical situations. The first microarray experiments demonstrated the promise of the technology by estimating patterns of gene expression and showing these patterns agreed with prior knowledge of the systems under study. More recent uses of microarrays for gene expression profiling, genotyping, mutation detection, and gene discovery are leading to remarkable insights into the function of thousands of genes previously known only by their DNA sequence. This proposal will use microarray technology to pinpoint a specific gene or groups of genes (operons, regulons) involved in pathogenesis or virulence of four organisms, Mycobacterium tuberculosis, Chlamydia trachomatis, Neisseria gonorrhoeae and methicillin-resistant Staphylococcus aureus (MRSA). Information gained by these studies will be utilized to develop improved diagnostics which could potentially be used to predict the next emerging epidemic or virulent strain. Duration: Jan 2002-Jan 2005 Funding: Office of biotechnology and Science, Health Canada; $699,000 Sub-project title: Viral genomics Co-PI: Alberto Severinin, Nathalie Bastien, Michael Drebot, Yan Li Study description: We are developing expression microarrays for HSV-1 and HSV-2 that will detect differences of expression among clinical isolates with respect to viral and host genes. S Difference in expression may be used to identify virulence markers associated with different clinical presentation of HSV infection. Duration: March 2002 - March 2005 Funding: $617,760

Project Title: Growth of reovirus in microcarriers in serum-free medium PI: M Butler; co-PI: KM Coombs Project description: Use of virus for vaccine purposes requires large-scale production of the virus. Most cell lines used for such purposes are grown in serum-supplemented medium. Recent detection of prions (such as Bovine Spongioform Encephalopathy – “Mad Cow Disease”) puts such production practices at risk. The purposes of this research project, a collaborative project with Dr. M Butler, Department of Microbiology at the University of Manitoba, are twofold: 1) to more efficiently produce preparative quantities of a model virus (mammalian reovirus) in suspension microcarrier cultures (1L of microcarriers = ~100 standard tissue-culture flasks); and 2) to attempt to grow the host cells in serum-free media, to avoid potential BSE risks. Results from these studies, which indicate that ten-fold higher titres than normal can be obtained from Vero cells grown in serum-free media on microcarriers, have been published in 2 articles prior to this reporting period, as well as in Burgener et al., 2001. We, and Butler’s group, are currently examining the energy requirements of reovirus-infected cells grown in microcarriers to try to optimize virus production. Duration and Funding: This project received funding from the UMRGC during July 1999 – June 2000. Although it is currently unfunded, the project continues.

Project Title: HCV and needle-sticks in health care workers Co-investigator: Chester Morris Study description: As above Duration: 2001-2002 Funding: $10,000

Project Title: HIV prevention in the occupational setting PI: Chester Morris Study description: As above Duration: 2001-2002 Funding: Illovo Sugar Social Investment Fund; $10,000.

Project Title: Home based care for vulnerable population in Kibera, Kenya PI: Chester Morris Study description: As above Duration: 2003-2004 Funding: Stephen Lewis Foundation; $60,000

Study Project: Host Factors Required for Hepatitis C virus Replication PI: M. Carpenter Project Description: Virus replication has a profound effect on host cell physiology. Modulations to specific signal transdcution pathways, antiviral responses and cellular replication agendas can be significantly compromised during the infection cycle. We are characterizing changes incurred to the host cell proteome during hepatitis C virus (HCV) gene expression. Although no current efficient cell culture or small animal model system exists in which to grow HCV, the development of self-replicating RNA molecules containing either the full viral genome or the nonstructural proteins has recently been developed (replicons). We are examining both complete and pre- fractionated protein samples from naïve host human liver cells or the same cells supporting HCV replicons (and thus expressing viral proteins). Special focus has been placed on identifying changes to the host cell phosphoproteome because of its major function in signal transduction. Changes are being determined using a compbination of protein purification (prefractionation), high-resolution 2-dimensional gel electrophoresis mapping, difference in gel electrophoresis (DIGE) triple labeling of protein extracts and mass spectrometry. Duration: 2002-2003 Funding: Office of the Chief Scientist (Health Canada); $55,000

Project Title: Host-Pathogen Interactions PI: Frank Plummer Collaborator: David Knox Project description: Prevention of human disease due to microbial infection requires a clear understanding of the mechanism by which pathogens circumvent both the immune-mediated and direct cytopathic host defense systems. A complete description identifying the alterations to a “normal” host genome and proteome profile in response to infection would be the most obvious starting point. Gene expression profiling is currently being used to identify changes in cellular transcripts for a number of host-pathogen interactions including hepatitis C virus, coxsavckie B3 virus, and Salmonella. While these genomics studies represent good starting points, they must be complemented and validated by determining the actual changes to the host proteins. It is after all, the corresponding proteins, whose levels can not always be predicted by the transcript profile, and which are subject to post-translational modifications, protein-protein interactions and targeting to subcellular compartments, that are directly responsible for phenotype. Therefore, we aim to characterise, at the protein level, the effect pathogens have on host cellular environments. This will provide a fundamental understanding of the pathogenic process, an opportunity for biomarker discovery and aid in the design and development of rational therapeutics. Prion disease -Prions are novel infectious agents consisting largely or entirely of misfolded isoforms of a ubiquitous host-encoded glycoprotein, PrP. The disease related isoform, PrPres, contains no demonstrable genetic material and is derived from the host protein, PrP, by a posttranslational process. The conformationally changed protein is recognized biochemically by its partial resistance to digestion by proteinase K and its detergent insolubility. Prion strains, encoding distinct disease phenotypes, are associated with distinct PrPres conformations and patterns of glycosylation that can be distinguished by western blot analysis following limited proteinase K digestion. Prion diseases are transmissible by inoculation with, or dietary exposure to infected tissues both within and between species. The transmission of prion diseases between different mammalian species is limited by a “species barrier” which is influenced both by prion strain and differences in the in the amino acid sequence of the PrP in the inoculum and the host. One form of human prion disease, variant Creutzfeldt-Jakob disease (vCJD), is thought to result from human dietary exposure to cattle suffering from bovine spongiform encephalopathy. These diseases are invariably fatal, however, the absence of any genetic material associated with the infectious particle precludes the use of highly sensitive gene based technologies currently applied to the detection of other infectious agents. Definitive diagnosis requires postmortem examination. The lack of a screening test for the preclinical disease makes it impossible to know the extent of the human health threat posed by the disease. Current estimates of the number of people at risk range from several hundred to hundreds of thousands.  Diagnostic Assay – The recent demonstration that prion replication can occur in an apparently resistant host (Hill et al., 2000) raises the spectre of a significant number of people harbouring an infectious titre of prion protein while remaining disease free themselves. To protect the public from this conceivably severe health risk an assay for the preclinical disease needs to be developed. While differential steady state mRNA levels in the brain have been observed a marker in a more accessible tissue or body fluid must be identified in order to make wide spread screening practical. Hamster scrapie strain Sc237 will be used to inoculate CD-1 mice. At various points post-inoculation the blood and urine of these animals will be screened for the protein poducts of specific mRNAs found to be differentially expressed in the brain. In addition, the blood and urine proteomes will be screened for any differentially expressed protein using high resolution 2D gel electrophoresis and protein identification by Mass Spectrometry. Duration: 2002-2005 Funding: Canadian Biotech Strategy, Industry Canada, $233,000.

Project title: Hot Spot Mapping: a quantitative approach for the geographic selection of areas of heightened HIV transmission on the Mombasa to Kampala transport corridor PI: Dr Alan Ferguson CO-PI: Dr C. Morris, Prof. E. Ngugi Project description: PURPOSE: The Overall objective of this project is to bring about a reduction in the level of HIV and STIs among the affected populations. This will be done by conducting hot spot mapping from Mombasa, on the Kenya Coast, through the main transport arteries of Kenya, into Uganda and on to Kampala. The results will provide information base to be used to quantify the need for programmed interventions and provide a framework upon which integrated interventions, using the tested community-based models of the STD Project, may be applied. HOT SPOT MAPPING: Hot-spot mapping has been one of the techniques developed and utilized within the Project to define geographical areas in which increased sexual networking takes place. These geographic areas may be defined in the context of known areas of high sexual exchange such as transport corridors or may be associated with specific geographic areas of towns or cities where this type of exchange is facilitated. The Project, in either case, uses standard approaches and experienced facilitators to gain entry into the community of a hot-spot and, by progressive use of key informants and snowballing of the vulnerable groups, is able to produce a map of individual or groups of hot spots, indicating the key locations and points where sexual interaction is most intensive. These interactions are mapped within the confines of a GIS defined area and health related and social variables are linked to these coordinates. The geographic and social information garnered from these maps form the basis of a targeted HIV intervention. TARGET AREAS: The project will map all the main highways of Kenya and their extension into Uganda. The mapping is proposed in three segments: 1. Mombasa – Nairobi 2. Nairobi – Kisumu – Busia and Mau Summit – Eldoret – Malaba 3. Busia and Malaba – Kampala MAPPING PROCEDURES: It is proposed that the hot spot mapping is completed in three phases, each phase corresponding to the above sectors and each budgeted separately. The Project proposes to undertake mapping of the hot spots of each highway segment, using existing and novel techniques to provide the following outputs: a) Detailed maps of points on the three highway segments described above, in which high levels of transactional sex take place, b)Linkage of these geographic points to key indicators of transactional sex, e.g., numbers of bars, lodgings, private clinics, parked trucks, volume of beer sales, average patronage of bars at different evening hours, estimates of FSW numbers, condom distribution, c) Linkage of these geographic points to points of health service provision including clinics, hospitals, STI care, pharmacies and private practitioners, d) GIS mapping of individual hot spots including the key services noted above, e) Narrative summary of each hot spot, detailing its characteristics, the dynamics of transactional sex and the unmet need for STI/HIV prevention and care programmes Duration: November 2003-April 2005 Funding: DFID through Futures Europe; Kshs 14,201,657 (as well as support from CIDA) Project Title: Imaging of Infectious Disease Agents: high-resolution cryo-transmission electron microscope. PI: Tim Booth Study description: Electron microscopy (EM) is an essential tool for identifying microbial disease-causing agents. In addition, it is very useful for studying the structure of disease agents such as viruses, in order to learn basic scientific facts about how these agents function in infection (for example; how they interact with cellular receptors and the immune system) and cause disease. The most commonly used TEM specimen preparation technique has been negative staining. Three-dimensional structures may be derived from these images by using digital image processing. Such studies explain key components of the immunity and pathogenesis of viruses, and provide knowledge relevant to the prevention and treatment of infectious diseases. Cryo-electron microscopy techniques allow ultra-rapid freezing of biological macromolecules in a thin layer of vitreous ice. These vitrified-hydrated specimens are then examined in a transmission electron microscope equipped with a low-temperature specimen holder. This method gives much improved preservation of structural detail and hence better results and higher resolution. Duration 04/2002-04/2003 Funding: Office of the Chief Scientist Strengthening Scientific Capability and Infrastructure, public Security and Anti-terrorism. $1,200,000

Project Title: Immunogenetic and Immunoregulatory Basis for Mucosal Immune Responses to HIV-1 in Highly Exposed Uninfected Sex Workers PI: Frank Plummer Collaborators: Keith Fowke Study description: As above Duration: April 2004-March 2009 Funding: National Institutes of Health; $398,444 per year

Project Title: Immunogenicity and acceptability of intranasal polyvalent inactivated influenza vaccine. PI: Fred Y. Aoki Collaborators: Greg Hammond, Pam Orr, Neil Simonsen Project description: This was a phase II study of the immunogenicity and acceptability of inactivated trivalent influenza vaccine administered intranasally on proteosomes. Duration: July 2002-July 2004 Funding: IDB Pharmaceuticals; $92,177

Project Title: Immunoregulation of graft-vs-host disease PI: John Gartner Collaborator: Kent HayGlass Study Description: As above Duration: 2002-2003 Funding: MICH; $40,000

Project Title: Immunogenetic & immunoregulatory basis for mucosal immune responses to HIV-1 in highly exposed uninfected Kenyan women PI: Frank Plummer Study description: We propose to expand our studies of resistance to further characterize mucosal immune mechanisms potentially mediating HIV-1 resistance with the goal of understanding the correlates of protection against HIV. Duration: July 2004 – June 2009 Funding: NIH; $1,900,725 USD

Project Title: Inactivation/decontamination of human and animal bioterrorism agents and analysis of suspicious materials with mixed hazards PI: Stefan Wagener Study Description: As above Duration: 2002 Funding: CRTI $160,000

Project Title: Influence of antibiotics on normal flora PIs: George Zhanel, Daryl Hoban, Dr C Donskey (Cleveland) Study description: Study assessing the impact of antibiotics on normal colonic flora Duration: 2002-2004 Funding: Ortho-McNeil; $300,000

Project Title: Innate and adaptive immune strategies for the prevention and control of Foot-and-Mouth disease Virus. PI: Jody Berry Project Description: As above Duration: 2003-2005 Funding: CFIA Technology Development Grant.

Project Title: Inventory and feasibility study of for creating a National Culture Collection for Canada Co-Investigators: Government colleagues in 4 departments; Agriculture and Agri-Foods Canada, A. Levesque, lead; CFIA, K. Amoako, lead; DRDC, B. Kournakakis lead; PHAC, K Bernard, lead. Project Description: As above Duration: 2004 Funding: CRTI; $550,000 over 1 year

Project Title: Investigation of molecular mechanism(s) involved in the action of HIV-1 integrase during nuclear import of viral preintegration complex. PI: Xiaojian Yao Study description: The capacity to infect nondividing cells productively is one of the hallmarks that distinguishes HIV- 1 from oncoretroviruses. At molecular level, the ability of HIV-1 to infect nondividing cells, incluiding macrophages, has been attributed to the karyophilic properties of HIV-1 preintegration complexes (PICs). Indeed, several HIV-1 proteins including Matrix (MA p17gag), IN and Vpr, which contribute to the efficient viral PICs nuclear transportation, have been reported to have karyophilic properties. Moreover, a recent study revealed that the formation of DNA flap is also critical for HIV-1 PIC nuclear import and viral infection in both dividing and non-dividing cells. HIV-1 integrase (IN) plays multiple roles during HIV-1 replication. Interestingly, in contrast to MA and Vpr which contribute to HIV-1 nuclear translocation in nondividing cells, IN has been shown to participate in HIV-1 nuclear import in both dividing and nondividing cells. However, even though intensive studies including using HIV-1 proviral clones have been dedicated to elucidate the mechanism(s) involved in the IN action during HIV-1 nuclear import, the results have been controversial and two main questions remains to be addressed. First, what mechanism(s) is involved in IN contribution to HIV-1 nuclear import in both dividing and non-dividing cells? Second, whether HIV-1 IN contains NLSs specifically contribute to HIV-1 nuclear import in nondividing cells? The goal of this proposed study is to investigate the functional role and the mechanism(s) involved in HIV-1 IN participating in the nuclear translocation of viral preintegration complex. To reach this goal, several perspectives will be studied: 1) To define the sequence and structural requirements for HIV-1 IN nuclear localization and their contributions to HIV-1 nuclear import in both dividing and nondividing cells. Especially, whether IN harbors determinant(s) specifically contributing to HIV-1 infection in nondividing cells will be addressed. 2) To elucidate the mechanism(s) involved in HIV-1 IN participation in HIV-1 PIC nuclear translocation. The study will investigate how HIV-1 IN, along with MA and Vpr, to utilize the karyopherin / mediated nuclear translocation pathway and what is the determinant in HIV-1 IN to recruit MA into viral core nucleoprotein complex, thereby allowing the MA NLS to play its role during HIV-1 PIC nuclear import. 3) To explore the potential IN-interacting cellular proteins involved in HIV-1 nuclear import, we will use a function-based genetic selection system in yeast to screen anti-integrase peptides and find the amino acid motifs that may present in IN-interacting cellular factors. By using different approaches presented in this proposal, we believe that these studies will provide valuable information for the better understanding of the functional role of IN during HIV-1 nuclear import and viral replication. Duration: April 2003 – March 2006 Funding: CIHR; $229,812

Project Title: Investigation of the natural history of West Nile virus infection in patients with or at risk for progression to West Nile virus encephalitis and/or myelitis. PI: Fred Y. Aoki Co-Investigators: Greg Hammond and Pam Orr Project description: An observational study of the natural history of West Vile virus infection. CASG study No. 211 Duration: June 2004 onward Funding: NIAID

Project Title: An invitro evaluation of new antifungal agents for the management of blastomycosis PI: John Embil Study description: As above Duration: 2001-2002 Funding: Fujisawa Inc; $50,000; Pfizer $50,000

Project Title: In vitro Pharmacodynamic Modelling of Ertapenem vs. Streptococcus pneumoniae of Various Resistance Phenotypes Co-PI: Daryl Hoban and George Zhanel Project Description: To assess the pharmacodynamic potential of Ertapenem in plasma vs. ELF inhibiting the growth of multiple genotypes of Streptococcus pneumoniae. Duration: Funding: Pharmaceutical Industry; $33,310/year

Project Title: In vivo half life determinations of SCFv mAB XXX PI: Kent HayGlass Study description: As above Duration: 2000-2001 Funding: Novopharm Biotech $1,929

Project Title: Ketolide activity against molecularly characterized Macrolide-resistant S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: Abbott $50,000

Project Title: Laboratory train surveillance of hepatitis B and hepatitis C viruses PI: MR Dawood Project Description: We are participating in a national study to determine the strains of both hepatitis B (HBV) and Hepatitis C (HCV) viruses in acute infected patients in Canada. Acute cases of HBV patients as identified by the presence of Ab against HBV core IgM. Newly infected HCV patients were interviewed by public health nurses to determine their infection history. Based on patient’s history we were able to determine if it is an acute case. Genetic material from both viruses were amplified and genotype was determined. Duration: Funding: Health Canada; $40,100

Project Title: Left ventricular function in sepsis Co-PI: Bruce Light, Drs Mink, Bose, Jacobs Project description: As above Duration: 2001-2007 Funding: Manitoba Heart & Stroke Foundation; $40,000 (2001-2), $115,200 (2004-7)

Project Title: Linezolid activity against penicillin-resistant and multi-drug resistant S.pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: Pharmacia $25,000

Project Title: Lipid A antagonist infusion in patients with severe sepsis PI: Bruce Light Study description: As above Duration: 2003 Funding: Esai Medical; $27,500

Project Title: Manitoba anonymous unlinked SEN-V seroprevalence study PI: MR Dawood Project Description: SEN-V is a new virus that was isolated from patients that were suffered from hepatitis like illness after heart surgery that required blood transfusion. The patients were negative for other known hepatitis viruses but positive for SEN-V. Since the patients have no other known risk factors for SEN-V infection. We studied the prevalence of SEN-V among the general population in Manitoba. The study design was anonymous unlinked where the demographic information were removed from blood samples provided for other tests. Then the specimens were tested for the presence of SEN-V. Duration: Funding: Health Canada; $42,800

Project Title: Medical countermeasures against Ricin Project lead: Cangene Corporation (Wendy Johnson) Collaborators: Twinstrand Therapeutics, Defence R&D Canada Project description: Ricin has become a serious threat to civilian and military personnel for two main reasons: because it is one of the most toxic substances known (lethal in microgram quantities - by inhalation or injection) and because it is readily accessible in most parts of the world. Moreover, public concern about the ricin 'threat' is fueled by a total absence of effective antidotes. The objective of this CRTI-funded project is to develop a fast- acting and effective antidote to ricin intoxication. Ricin is a protein product of castor bean, a plant cultivated worldwide for its oil and as an ornamental. Millions of tonnes of castor bean are grown annually with major production in Brazil, India and China. The ricin toxin comprises roughly 5% by weight of the bean and the protein is readily extracted from beans using low-tech methods. Ricin has an ignoble history:

 it has been used in assassination in the UK;  it has been found in the possession of white-supremacists in the US;  authorities in the UK recently uncovered ricin in the possession of members of a terrorist cell;  a flask containing ricin was recently found in a Paris train station;  elements of the former Ba'athist regime in Iraq are suspected of developing ricin-based weapons.

Ricin is toxic whether it is delivered as an aerosol, by injection or through ingestion. However, the greatest threat to civilian and military personnel - and public confidence - is a ricin attack involving aerosolized forms of the toxin. Twinstrand Therapeutics, Cangene Corporation, and the Defense R&D Canada (DRDC) will use complementary expertise to generate antibody-based countermeasures to ricin. A countermeasure strategy involving the passive immunization of afflicted personnel is preferred to vaccination because of the rapid clearance kinetics of the ricin toxin (a ricin attack would have to be precisely anticipated days or weeks in advance for ricin vaccine or ricin booster to be effective). Twinstrand Therapeutics will have primarily responsibility for developing non-toxic ricin 'antigens' with similar or identical immunogenic properties to the authentic toxin. Cangene will use the Twinstrand antigens to generate polyclonal caprine, and monoclonal human antibodies. The DRDC will be primarily responsible for evaluating the effectiveness of different antibody preparations in animal challenge experiments. The structure and activity of ricin antigens is key to the success of the project. Antibodies prepared using protein fragments, chemically modified or denatured forms of ricin are known to cross-react poorly with the natural toxin; observations that indicate a full-sized and properly folded protein is necessary for effective immunization/antibody screening. Twinstrand has developed a unique expertise in the design, expression and purification of recombinant ricin-like molecules and has patented methods for controlling/ attenuating the cytotoxic activity of these proteins. Importantly, the proteins made by Twinstrand are full-sized and properly folded ricin antigens. Animal dosing for immunization can be optimized because the antigens are non-toxic. Recombinant ricin antigens will be made using a Pichia pastoris yeast expression according to Good Manufacturing Practices as defined by the US FDA. Cangene Corporation is a commercially successful leader in the development of antibody-based therapeutics. The company has relevant, proprietary, expertise in the preparation of hyperimmune products involving; 1) stimulation of the immune system, 2) extraction, purification and decontamination of immunoglobulin from hyperimmune sera, and 3) formulation of immunoglobulins for parenteral use in humans. Cangene also has relevant expertise in the development of human monoclonal antibodies. Therefore, Cangene will apply its expertise to produce hyperimmune ricin antisera in goats and simultaneously screen antibody phage libraries for ricin-neutralizing monoclonals. This two-pronged approach will facilitate the rapid generation of an antidote based on caprine polyclonal antibodies while enabling the long term development of an antidote based on human monoclonals. The effectiveness of polyclonal and monoclonal antibodies will be determined at the DRDC in Suffield where there are unique facilities for animal challenge experiments and there is a significant background in the handling and testing of ricin. At the conclusion of the project, the consortium will deliver a ricin antidote suitable for scale-up, human testing, commercialization and protocols for the use of the antidote. Duration: October 2003 onwards Funding: CRTI $1,739,900

Project Title: Medi-Safe Fine Lumen Irrigator Study PI: Michelle Alfa Duration: 2000-2001 Funding: Medi-Safe Inc, $12,000

Project Title: Microarray detection of bioterrorism bacterial agents PI: Amin Kabani Project description: As above Duration: As above Funding: Office of Chief Scientist

Project Title: Microarray Expression Profiling in Prion induced Neurodegenerative Diseases to develop novel molecular typing strategies for classifying prion diseases and to identify surrogate markers of infection. PI: Stephanie. Booth Co-PI: R.L. Somorjai Collaborators: C. Bowman, R. Baumgartner, G. Glazner, K. Qingzhong Project Description: The central objective of this project is to develop a novel molecular typing strategy for the classification of prion induced neurodegenerative disease using gene expression profiling and to identify surrogate biomarkers that could be used to identify prion infection. Prion diseases comprise a distinct group of fatal transmissible neurodegenerative diseases of animals and humans. They include Creutzfeld-Jakob disease (CJD) and kuru in humans, and bovine spongiform encephalopathy (BSE) and scrapie in animals. As the prion agent has no genome, the term “strain” given to isolates of the infectious agent is something of a misnomer. Transmissible spongiform encephalopathies (TSEs) strains are distinct varieties of prion, which cause different incubation times, and deposition of the misfolded isoform of prion, PrPSc, in the brains of infected individuals. Transmission of TSEs across species is well-documented, and was recently brought to the forefront by the BSE epidemic in the UK and the subsequent emergence of a novel form of CJD called variant CJD (vCJD), believed to be acquired from ingestion of BSE infected cattle. BSE prions have strain characteristics identical to prion isolates from human cases of variant CJD (vCJD. Strain characteristics are often maintained even after passage in alternate host species. Characterization of different TSE strains is extremely important for the diagnosis and surveillance of disease, for example, to track the source of an outbreak, and to identify the incidence of cross- species transmission of TSEs. Diagnostic methodologies for prion diseases are for the most part based on the detection of the disease-specific isomer of the prion protein. The large-scale analysis of gene expression creates the possibility to define a disease based on the coordinated action of a group of genes. Thus, biomarker selection can be performed at high throughput and information on disease susceptibility and prognosis can be obtained. Similarly, improved diagnosis, and potential therapeutic targets can be gleaned. In addition it should be possible to use gene-expression profiles for biomarker selection and to gain an insight into the molecular pathogenesis of prion induced neurodegeneration, poorly understood at present. We have identified a group of genes that are differentially expressed in mice infected with different strains of prion at different times post infection. We have developed a strategy for the creation of molecular classifiers with strong discriminatory potential for these strains. Further work is underway to identify the molecular mechanisms of neuronal death in prion induced neurodegenerative disease. We are using microarray profiling in cell culture models of prion infection and in primary neurons. Duration: 2003-2005 Funding: Health Canada’s Health Canada's Blood Safety Program; CIHR (2003 – 2005) $120,000; MMSF (2003 – 2004) $40,000; (2004 – 2005) $20,000; Office of the Chief Scientist, Health Canada (2003-2004) $32,690

Project Title: Molecular Characterization of Macrolide Resistant S. pneumoniae in Canada. Co-PI: Daryl Hoban and George Zhanel Project Description: To determine the molecular mechanisms of macrolide resistance in isolates of S. pneumoniae isolated in Canada. Determine via PCR the prevalence of erm and mef genes in all macrolide non-susceptible S. pneumoniae isolated in Canada in 2004 from the 2004 CROSS study. Duration: 12 months Funding: Pharmaceutical Industry; $30,000/year

Project Title: Molecular characterization of a novel neurotropic human reovirus PI: KM Coombs Project description: Reovirus gets its name from “respiratory” and “enteric” (the routes by which it normally infects hosts), and “orphan”, to indicate it is not normally associated with human disease. In November 1997, an 8-week-old infant girl presented with a three week history of active varicella complicated with E. coli sepsis, persistent oral thrush, hypoalbuminemia, intermittent fevers with temperature spikes to 39 C, diarrhea and feeding intolerance and was hospitalized at the Children’s Hospital (Winnipeg, Manitoba) for investigation of congenital immune deficiency. A potentially novel reovirus (designated T2W because it appeared to belong to the “type 2” group of reovirus) was isolated from CSF and demonstrated a novel RNA pattern in gels. This initial observation was written as a case report (Hermann et al., 2004) and follow-up studies are underway to molecularly characterize the agent. Sequencing studies of 3 of the agent’s 10 gene segments indicate this is a novel reovirus clone, although 2 of the genes bear striking similarity (>99%) to reoviruses in the “type 1” group. We plan to sequence another 2 genes of the T2W isolate to allow further molecular comparisons with currently known clones. Future plans include determination of T2W’s growth potential in various cell lines including those of neuronal origin. Duration and Funding: This project, although currently not funded, has been underway since 2000

Project Title: Molecular characterization of novel infectious agents PI: KM Coombs; Collaborators: PR Hazelton, P vanCaseele Project description: It has been estimated that each year there are more than 2 billion episodes of gastroenteritis, leading to millions of deaths and incalculable societal costs. Viruses are the primary cause of infectious gastroenteritis, with rotavirus being the leading cause. However, it is important to note that the second leading cause of viral gastroenteritis are caused by unknown agents. Thus, we are involved in the discovery of, and molecular characterization of, novel viral agents. In addition to standard electron microscopic detection of viruses in clinical samples submitted through the Cadham Provincial Laboratories, we have developed a novel ultra- sensitive immunological assay to determine whether new agents have a DNA or an RNA genome. Potentially novel agents also are being detected by molecular biologic methods. Results from this project have been reported at a variety of scientific conferences (including, during this reporting period Hazelton et al., 2001, Roche Light- Cycler Conference, Winnipeg) and has been submitted. Duration and Funding: This project, although currently unfunded, received funding from the Dr. P Thorlakson Foundation from July 1996 – June 1997 and from the UMRGC from July 1999 – June 2000. Project has been underway since 1991

Project Title: Molecular detection of weaponized arenaviruses Collaborator: Heinrich Feldmann Project description: The objective of the proposed research is to design, develop and evaluate molecular assays to permit rapid and easy detection by real-time RT-PCR of weaponized arenaviruses (AVs) used in a bioterrorism context. Weaponizable AVs include 5 viruses classified in the Category A Pathogen List as defined by the CDC (BSL-4 agents), and all potentially engineered AVs consisting of mutated and/or recombinant and/or reassortant strains. Little genomic data is available for AVs. Nevertheless, genetic recombination and reassortment have been observed in natural or experimentally produced AVs. Therefore, a knowledge of the complete genomic sequence of all AVs is the cornerstone of the detection of weaponized AVs. To distinguish BSL-4 AVs from other AVs, of which at least four are naturally circulating is North America and potentially infecting humans, and to detect chimeric engineered AVs it is critical to determine the complete sequences ofBSL-4 AVs, but also BSL-2/3 AVs. The realization of this project requires - an extensive experience in genomics of RNA viruses, - expertise in the design/evaluation process of diagnostic assays of human pathogens, - a perfect knowledge of arenavirus genetics, - an undisputable working experience of BSL-4 pathogens, and - the immediate availability of a BSL-4 laboratory. The molecular assays will be developed at the Unité des Virus Emergents in Marseilles, France and evaluated comparatively and experimentally in the BSL-4 facilities in Winnipeg, Ontario, Canada. The ultimate goal is to propose a ready-to-use diagnostic kit for the detection and characterization of weaponized arenaviruses. Duration: 2002-2004 Funding: National Institute of Allergy and Infectious Diseases (NIAID), Special Emphasis Panel : $ 50,000 (US)

Project Title: Molecular epidemiology of biothreat agents PI: Michael Mulvey Study description: As above Duration: 2003-2008 Funding: CRTI $1,860,835

Project Title: Molecular epidemiology of Blastomycoccis in Manitoba Co-PI: John Embil, George Zhanel, Daryl Hoban Study description: Study assessing the molecular epidemiology of Blastomycosis in Manitoba Duration: 2003 Funding: MMSF $10,000

Project Title: Molecular epidemiology of HAV in Canada PI: Anton Andonov Project Description: As above Duration: Ongoing Funding: Public Health Agency of Canada

Project Title: Molecular epidemiology of vancomycin-resistant entercoccci in North America PI: George Zhanel, Daryl Hoban, Jack Johnson Study description: Study assessing the molecular epidemiology of VRE in North America Duration: 2003 Funding: Procter and Gamble $280,000

Project Title: Molecular mechanism of action and resistant to an new antibiotic G 1 Co-PI: George Zhanel, Daryl Hoban, Study description: As above Duration: 2003 Funding: Roche $35,000

Project Title: Molecular resistance to antibiotics Co-PI: George Zhanel, Daryl Hoban, Kris McNichol Study description: Assessment of the cellular and molecular mechanism of action of magainins Duration: 2003-2004 Funding: Micrologix $30,000

Project Title: Monte Carlo simulation of fluoroquinolones Collaborators: George Zhanel, Daryl Hoban, Dr G Drusano (New York) Study description: A study assessing the optimal sampling against resistant pathogens Duration: 2003-2004 Funding: BristolMeyersSquibb; $26,000

Project Title: Mother-Child HIV Transmission and Pediatric AIDS in Nairobi Kenya CO-PIs: Joanne Embree, Frank Plummer, L Gelmon Project Description: This is a long standing cohort study conducted with the University of Nairobi in Nairobi, Kenya. The goal is to determine epidemiologic, immunologic, and virologic factors that enhance or protect against maternal transmission of HIV from an infected mother to her child. Maternal-infants pairs were enrolled into the cohort from January 1986 through August 2000 and followed prospectively. Both HIV seropositive and seronegative mother-infant pairs were enrolled. Infants born to seropositive mothers were followed to determine whether they acquired HIV from their mother perinatally or through breast milk or whether they escaped infection. Until recently, antiretroviral agents for the prevention of perinatal HIV transmission or for the treatment of HIV/AIDS were not available or accessible. This situation is being rectified and the study is being modified to address the issues related to the implementation of preventive and therapeutic interventions in this setting. Duration: 1986 and ongoing Funding: Previously EEC, CHRF, NIH, IDRC, NHRDP, MRC, and CIHR grants, Currently - CIHR Core Group Grant Funding (October 2001 – September 2006) is supporting core studies. $1,749,250

Study Project: Mother-risk Program related to Perinatal HIV Transmission to Canadian Children PI: King SS, Project Collaborators: Singer J, Forbes J, Lapointe N, Samson L, Vaudry W, Embree J, Orr P Project description: This is an ongoing study of the maternal risk factors associated with transmission of HIV from infected mothers to their children in Canada and the effects of the use of antiretroviral agents to prevent transmission. Duration: Ongoing Funding: Hospital for Sick Children, Toronto Project Title: A Multicentre, Double-Blind, Comparative, Randomised Study to Evaluate the Efficacy and Safety of Micafungin (FK463) versus Liposomal Amphotericin B (Ambisome) in the Treatment of Invasive Candidiasis and Candidaemia Co-PI: Eric Bow Project Description: As above Duration: July 2003-July 2004 Funding: Fujisawa GmbH, Protocol FG-463-21-08

Project Title : A Multicentre, Double-Blind, Randomised, Phase 2 Trial Comparing Pegfilgrastim with Filgrastim as an Adjunct to Chemotherapy for Acute Myeloid Leukaemia Co-investigator: Eric Bow Project description: As above Duration: May 2003-March 2004 Funding: Amgen Development Europe, Protocol 20020163; $15,500

Project Title: Mutant prevention concentration of fluoroquinolones versus S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: Janssen-Ortho $30,000

Project Title: Natural history and treatment of Acute Exacerbations of Chronic Bronchitis (AECB) Co-PI: George Zhanel, Charlie Chan Study description: As above Duration: 2002-2005 Funding: Janssen-Ortho $660,000

Project Title: National real-time network for identification of bioterrorist agents Co-investigator: Michael Mulvey Study description: As above Duration: 2003-2004 Funding: CRTI $1,330,000

Project Title: Near-infrared, thermal and magnetic resonance imaging in diagnostic and treatment of cardiac ischema and infarction PI: Xi Yang Study description: As above Duration: 2004-2007 Funding: CIHR/NRC; $1,048,080

Project Title: Networks and infectious disease: social and molecular factors affecting transmission of hepatitis C and HIV among injection drug users. PIs: John Wylie and Ann Jolly. Co-applicants: Keith Fowke Study description: Our research program focuses on sexually transmitted and bloodborne pathogens among high risk populations. We combine traditional epidemiologic approaches, with the individual as the unit of analysis, with social network analysis and molecular epidemiology. The current proposal deals with the factors associated with the bloodborne transmission of Hepatitis C (HCV) and HIV among injection drug users (IDU) and is based on data that we collected from a recently completed pilot study. The strength of social network analysis is its focus on the interactions and behaviours which occur between individuals; the key analytic point for research on the epidemiology of pathogens whose transmission depends on intimate and complex social behaviours between two or more people. Molecular investigations have recently furthered epidemiological investigations as an independent means of verifying the accuracy of social data, and for the identification of large scale network patterns not readily discernible using social data alone. Given the relatively recent emergence of these approaches into the mainstream of infectious disease epidemiology, social network and molecular investigations have not yet been extensively applied to questions related to the transmission patterns of sexually transmitted and bloodborne agents. We are using these approaches to address the following specific aims: Aim 1. Analyze the social context of syringe sharing among IDU. We will test the hypotheses that intimacy, behavioural norms and actions, and/or individual personality traits govern the likelihood that two people will share a syringe. Aim 2. Correlate social network variables with the seroprevalence of HCV and HIV. The hypothesis here is that the risk of HCV and HIV transmission is a function of an individual's behaviours, social network characteristics, or both. Aim 3. Analyze the molecular epidemiology of HCV within social networks. We hypothesize that networks of IDU segregated along socioeconomic or behavioural lines can be identified with molecular HCV genotype data. Aim 4. Correlate immune system status with social behavioural data. The hypothesis is that positive social support will correlate with variation in molecular markers or functional activity of immune system components. Aim 5. Construct and analyze sociometric networks of IDU. We will gauge the feasibility and acceptance among our study population of a specific type of network analytic approach. A questionnaire will be administered to IDU (injection drug use in previous 6 months) as well as collection of a blood sample. In addition to individual demographics and behavioural data, each participant will list their personal (egocentric) social network (anonymously, by first name or initials as was done in our pilot study) and be asked questions about the demographics and behaviours of the people listed. The questionnaire data will be used for the syringe sharing analysis of aim 1, and combined with the molecular analyses for aims 2-4. Our units of analysis will be either at the level of the individual, or based on dyad relationships between a given study participant and a specific member of their network. Blood specimens will provide seroprevalences for aim 2; HCV genotype data for aim 3; and immune system data for aim 4. Statistical analysis for the project will be by multivariate logistic or linear regression techniques, as appropriate. The project is budgeted for a 3-year time frame. The proposal is innovative in several ways. It will result in the most extensive database of IDU social networks in Canada and will be the first to conduct a detailed analysis of the social context of syringe sharing among a Canadian IDU population using a network approach. It is the first to combine HCV seroprevalence data with social network data to refine our understanding of risk behaviours for this pathogen. The molecular epidemiology of HCV within networks has also not been extensively analyzed, and the effect of social support on immune system function in IDU has not previously been addressed. This project will illustrate to a Canadian and international audience the type of network studies that can productively be entered into with IDU and will provide results applicable to the development and refinement of public health programs for the prevention and control of bloodborne infections among IDU. Duration: 2002 - 2005 Funding: CIHR; $360,000

Project Title: New technologies or surveillance of biowarfare agents and identification of genetically engineered virulence genes Collaborator: Michael Mulvey Study description: As above Duration: 2003-2007 Funding: CRTI $2,472,750

Project Title: North American Urinary Tract Isolate Collaborative Alliance (NAUTICA) CO-PI: George Zhanel and Daryl Hoban; Collaborators Lindsay Nicolle, Godfrey Harding Project Description: To assess the prevalence of antibiotic resistance of Quinolone Resistant E.coli in outpatient midstream urinary isolates from major United States and Canadian Medical Centres. A total of 30 geographically dispersed study sites (labs) will be recruited representing all regions of the United States. A total of 10 geographically dispersed study sites (labs) were recruited representing all regions of Canada. At the reference lab all isolates obtained will have susceptibility profiles determined. Duration: Ongoing Funding: Pharmaceutical Industry; $250,000/year

Project Title: Nosocomial Pneumonia in PICU PI: Stephanie Black, Co PI :Evelyn Lo; collaborators: John Segreti Study description: Prospective case-control study on risk factors for development of nosocomial pneumonia in the PICU at Rush-Presbyterian Medical Center in Chicago. Patients identified on census if admitted greater than 48 hours in PICU and fulfill NNIS definition for a nosocomial pneumonia. Data gathered regarding various risk factors for development of pneumonia. Cases matched by age and duration of admission. Duration: Ongoing Funding: infection control QI project

Project Title: Novel Technology for Reprocessing Flexible Endoscopes PI: Dr. Mohammed Labib, Novaflux, Princeton, NJ. Collaborators: Michelle Alfa Project Description: As above Duration: 2002-2004 Funding: NIH $80,000 US

Project Title: An Open Label Non-Comparative Protocol for the Emergency Use of Voriconazole in Patients with Life-Threatening Invasive Mycoses Who Are Failing on Currently Available Antifungal Agents. Collaborator: Eric Bow Project Description: As above Duration: 1999-present Funding: Pfizer Protocol A1501050

Project Title: Open-label study of recombinant activated protein C in severe sepsis PI: Bruce Light Study description: As above Duration: 2002 Funding: Lilly Pharmaceuticals; $224,000

Project Title: Osteoporosis and HAART in Canadian Women: a multicenter trial Co-Investigator: Chester Morris Study description: As above Duration: 2001-2003 Funding: Canfar; $180,000

Project Title: Outcomes of Manitoba patients with pyelonephritis treated with sulfonamides versus fluoroquinolones PI: George Zhanel, Daryl Hoban, Anita Carrie Study description: As above Duration: 2003 Funding: Janssen-Ortho $36,500

Project Title: Papilloma virus vaccine for prevention of infection by same PI: Fred Y Aoki Collaborators: Multiple Project Description: We were one of multiple centres involved in the phase II placebo-controlled evaluation of an HPV-16/18 vaccine for prevention of infection by these serotypes in susceptible women. Duration: Feb 2000 – Jan 2001 Funding: SmithKline Beecham; Study SKB 269814/005: $19,788

Project Title: Pathogenesis and antibiotic resistance in Campylobacter PI : Dr. D.E. Taylor Co-PI : Dr. Lai-King Ng Project description: This research will investigate the ways in which the foodborne bacteria, Campylobacter jejuni, transmits antibiotic resistance and other factors which allow it to cause disease in humans. The bacteria is found in animals (chickens, cattle) and in the environment. This research project will isolate the bacteria from chickens and cattle and compare them to bacteria isolated from people who have had food-poisoning with diarrhea. All of the bacteria will be typed and tested for antibiotic resistance and ability to invade cells in culture. Sophisticated techniques such as DNA microarray chips will be used to find the genes responsible for causing disease in humans. The identification of these genes will provide targets for developing new drugs and vaccines to reduce or eliminate the presence of these organisms in food. Duration: October 2003-September 2006 Funding: CIHR; $427,000

Study Project: Perinatal HIV Transmission to Canadian Children Co-PI: King SS, Project Collaborators: Singer J, Forbes J, Lapointe N, Samson L, Vaudry W, Embree, J Project description: This is an ongoing surveillance program of the incidence and changing epidemiology of the transmission of HIV from infected mothers to their children in Canada. Duration: Ongoing since 1993. Funding: Health Canada

Project Title: Pharmacodynamics of antibiotics versus urinary pathogens PI: George Zhanel, Daryl Hoban, Ayman Noreddin Study description: As above Duration: 2003-2004 Funding: Procter and Gamble $150,000

Project Title: Pharmacodynamics of AR-100 Co-PIs: George Zhanel and Daryl Hoban Study description: Pharmacodynamics of a new aminopyrimidine vs MDR organisms Duration: 2003-2004 Funding: Arpida; $60,000

Project Title: Pharmacodynamics of B-Lactams against penicillin-resistant S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: Bristol-Myers Squibb $16,000

Project Title: Pharmacodynamics of B-lactams vs multidrug resistant S. pneumoniae Co-PIs: George Zhanel, Daryl Hoban Study description: Assessment of the optimal B-lactam treatment of MDR S. pneumoniae Duration: 2003-2004 Funding: BristolMyersSquibb; $16,000

Project Title: Pharmacodynamics of fluoroquinolones versus H. influenzae PI: George Zhanel, Daryl Hoban, James Karlowsky Study description: As above Duration: 2003 Funding: Janssen-Ortho $42,000

Project Title: Pharmacodynamics of fluoroquinolones versus multi-drug resistant S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: Janssen-Ortho $56,000

Project Title: Pharmacodynamics of fluoroquinolones vs ParC mutants of S. pneumoniae PI: George Zhanel, Daryl Hoban, Dr D. Nicolau-Hartford Study description: As above Duration: 2002-2005 Funding: Ortho McNeill; $92,000

Project Title: Pharmacodynamics of gemifloxacin versus fluoroquinolone resistant S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: GlaxoSmithKline $56,000

Project Title: Pharmacodynamics of ketolides vs S. pneumoniae and H. influenzae PI: George Zhanel and Daryl Hoban Study description: As above Duration: 2003-2005 Funding: Aventis; $92,000

Project Title: Pharmacodynamics of macrolides/ketolides against molecularly characterized Macrolide-resistant S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2004 Funding: Abbott $50,000

Project Title: Pharmacokinetic study of cyclosporine or mycophenolate mofetil with or without a single concurrent dose of Oseltamivir in renal transplant patients. PI: Fred Y. Aoki Co-Investigators: Dr. H. Lam, J. Jeffrey, D.S. Sitar Project description: This is a descriptive pharmacokinetic study seeking an interaction between the anti-influenza drug, Oseltamivir, and cyclosporine and mycophenolate, which are important immunosuppressive agents administered to patients with renal transplant. Duration: 2003 onwards Funding: Hoffmann-La Roche Limited; $75,000

Project Title: The pharmacokinetics and pharmacodynamics of meropenem PIs: Dr. Robert Ariano, Dr. Sheryl Zelenitsky, Dr. Godfrey Harding Project description: As above Duration: 2002 Funding: AstraZeneca; $39,500 Project Title: Phase I/II randomized, placebo-controlled trial of the safety and efficacy of intravenous immunoglobulin G containing high anti-West Nile virus antibody titre in patients with or at high risk for progression to West Nile virus encephalitis and/or myelitis. PI: Fred Y. Aoki Co-Investigators: Greg Hammond and Pam Orr Project description: The University of Manitoba is one of multiple centres participating in this NIAID-sponsored efficacy trial; CASG study No. 210: Duration: June 2004 onwards Funding: NIAID;

Project Title: Phase 2a Randomized Study of Topical Resiquimod for Treatment of acute recurrent HSV. Principal Site Investigator: Dr. F. Aoki; Coinvestigator: Dr. P. Orr Duration: 2004. Funding: Royal Mount Pharma

Project Title: Phase II multi-centre, randomized, placebo-controlled trial to evaluate the safety and efficacy of topical RM100 12.5% and 19% cream for treatment of recurrent Herpes simplex labialis in the pre-lesional stage. PI: Fred Y. Aoki Collaborators: Greg Hammond and Pam Orr Project description: As per the title, individuals were randomized to receive on of the three treatments. They took a swab at the first symptom of a recurrence of their cold sore for HSV culture and PCR and initiated topical therapy. They reported to the research staff within 24 hours of initiation of therapy. Duration: December 2003 onwards Funding: Royalmount Pharma; $50,397

Project Title: A Phase 3 double-blind placebo-controlled randomized multicenter study of the efficacy of an HPV-16/18 VLP vaccine compared to Hepatitis A vaccine as control in the prevention of persistent HPV-16 or HPV-18 cervical infection and neoplasia in healthy females 15-25 years of age. Principal Site Investigator: Dr. F. Aoki; Coinvestigator: Dr. P. Orr Study description: As above Duration: 2004 Funding: GlaxoSmithKline

Project Title: A phase 3 Double-Blind Placebo-Controlled Trial of Long Term Therapy of HSV Encephalitis: An Evaluation of Valacyclovir Principal Site Investigator: F. Aoki; Coinvestigator: P. Orr, Study description: As above Duration: 2004. Funding: NIAID, NINDS, NIH.

Project Title: A Phase III, Double-Blind, Randomized, Multi-Centre Study of the Safety and Efficacy of Anidulafungin vs. Fluconazole in the Treatment of Patients with Candidemia and Other Forms of Invasive Candidiasis and Prevention of Complications Co-investigator: Eric Bow Project Description: As above Duration: May 2003-September 2004 Funding: Vicuron Pharmaceuticals, Protocol VER002-9; $77,520.00 Project Title: Pleconaril for prevention of the common cold (Protocol 843-062). PI: Fred Y. Aoki Collaborators: Greg Hammond, Pam Orr, Neil Simonsen Project Description: A placebo-controlled trial of Pleconaril prophylaxis for preventing the common cold in otherwise healthy adults. Duration: Winter and Spring 2001-2002 Funding: ViroPharma Incorporated; $216,248

Project Title: Pleconaril URI treatment study (Protocol 843-043). PI: Fred Y. Aoki, Collaborators: Greg Hammond, Pam Orr, Neil Simonsen Project Description: This is a placebo-controlled trial to evaluate the therapeutic efficacy of Pleconaril, a specific inhibitor of rhinoviruses replication, in the treatment of individuals with the common cold in a placebo-controlled design. Duration: Winter 2000-2001 Funding: ViroPharma Incorporated; $74,570

Project Title: Prevalence of SEN-V among high and low risk individuals PI: MR Dawood Project Description: SEN-V is a new virus that was isolated from patients that were suffered from hepatitis like illness after heart surgery that required blood transfusion. The patients were negative for other known hepatitis viruses but positive for SEN-V. Since the patients have no other known risk factors for SEN-V infection. It was proposed that the virus can be transmitted by blood transfusion. We studied the prevalence of SEN-V among high and low risk individuals in Manitoba. Duration: Funding: Health Canada; $42,800

Study Title: Prion-induced Neurodegeneration and Oxidative Stress PI: J. David Knox (PI), Collaborators: Ron E. Mitchel (Atomic energy of Canada Ltd.), Stephanie Czub (Canadian Food Inspection Agency), Douglas R. Boreham (McMaster University). Study description: Transmissible spongiform encephalopathies are untreatable, uniformly fatal degenerative syndromes of the central nervous system. A hallmark of these diseases is the accumulation of misfolded isoforms of the host-encoded prion protein (PrP) accompanied by neuronal death and glial cell proliferation. An etiologic association between the accumulation of the misfolded proteins and neurodegeneration is supposed, but the mechanism(s) remain unknown. The cellular form of the PrP is a highly conserved cell surface glycoprotein expressed by a broad range of cell type and in particular by neuronal cells. The function of PrP is unknown, however, biochemical studies and PrP knock-out mice have demonstrated that PrP plays a role in the oxidative state of the cell. During the course of TSE disease progression PrP undergoes a conformational modification into a protease-resistant isoform called PrPres. The loss of PrP function correlates with increased oxidative stress suggesting that loss of PrP function is indirectly responsible for neuronal death. Previously, we have demonstrated that in C57/BL6 mice intracerebrally infected with the prion strain ME7, four exposures to 500 mGy radiation ≤50 days post inoculation (dpi) significantly prolonged survival. It is our hypothesis that reactive oxygen species produced by low dose ionizing radiation induces an adaptive response that activates an endogenous anti- oxidant protective mechanism that inhibits the rise of oxidative stress induced by prion infection and delays the onset of the disease. We have also tested the effectiveness of delivery of the proven 500 mGy exposures at times later than 50 dpi. These experiments demonstrated that radiation treatments administered at 100 or 120 dpi do not significantly influence disease progression and when administered ≥140 an adverse effect is observed. We postulate that the oxidative stress imparted by the infection at these later stages of disease progression has already fully activated the endogenous anti-oxidant systems and that at day 140 dpi the additional reactive oxygen species generated by the radiation treatments exacerbate the situation.We propose to further test the hypothesis that the neuronal loss observed in TSE diseases is due to oxidative stress, by using transgenic mice (MT-rGH) known to produce elevated endogenous levels of free radicals and a proven system for the study of neural degeneration. Our hypothesis would predict the following: 1) The MT-rGH mice, due to a higher basal level of oxidative stress, will succumb more quickly to prion infection. 2) The MT-rGH mice will not benefit from radiation treatment because their high basal level of oxidative stress will have already induced their endogenous anti-oxidant systems. 3) In contrast, radiation treatment will result in the preemptive induction of endogenous anti-oxidant systems and prolonged survival in the non-rGH expressing isogenic littermates. 4) An anti-oxidant diet, already shown to reverse the neural degeneration observed in MT-rGH mice, will offset the increased reactive oxygen species generation caused by prion infection and protect both normal and MT-rGH mice from disease progression. In addition monitoring survival as an end point, powerful gene based technologies and biochemical tests will be employed throughout the course of the disease to identify the onset of oxidative stress and the activation of anti-oxidant systems. These experiments will contribute to our understanding of the mechanism(s) contributing to neuronal loss in this class of poorly understood diseases and could have high impact on our ability to manage these afflictions. Duration: Started 2004.

Project Title: Prospective multi-province surveillance for antimicrobial resistant E.coli in drinking and recreational source waters: Impact on humans and the environment. Co-Investigator: Michael Mulvey Study description: As above Duration: 2003-2006 Funding: CIHR; $560,000

Project Title: Protection and pathology in chlamydial infection PI: Xi Yang Study description: As above Duration: 2002-2005 Funding: CIHR/MHRC $235,000

Project Title: Purification and characterization of SARS CoV structural proteins. PI: Anton Andonov Project Description: As above Duration: Ongoing Funding: Public Health Agency of Canada

Project Title: PulseNet Canada PI : Dr. Lai-King Ng Project description: To facilitate the implementation of PulseNet Canada to allow real time inter-laboratory exchange of data. Duration: Jan 2003 – Mar 2003 Funding: Office of the Chief Scientist, Health Canada; $7,500

Project Title: A randomized, controlled trial of male circumcision to reduce HIV incidence in Kisumu, Kenya. Co-PIs: Stephen Moses (CIHR); Dr Robert C. Bailey (NIH) Study description: The principal objective of this study is, through a randomized controlled trial design, to assess the effectiveness of male circumcision in reducing HIV incidence. Duration: April 2001 – March 2006 Funding: CIHR; $1,816,700 Can; NIH $6.2 million USD

Project Title: Randomized Controlled Trial of Posaconazole (SCH56592) vs. Standard Azole Therapy for the Prevention of Invasive Fungal Infections Among High-Risk Neutropenic Patients Co-investigator: Eric Bow Project Description: As above Duration: April 2002, ongoing Funding: Schering Canada Inc., Protocol P01899-17; $74,220.00

Project Title: Randomized Double-Blind Placebo-Controlled Multicenter Study of Efficacy and Safety of a Patient-Initiated 1-day Treatment with Famciclovir 1000 mg bid for Recurrent Genital Herpes Infection in Immunocompetent Patients. Principal Site Investigator: F. Aoki; Coinvestigator: P. Orr Study description: As above Duration: 2004. Funding:

Project Title: Randomized, double-blind, controlled study to evaluate the safety & immunogenecity of MEDI-516 with MF59C, an E. coli pilus vaccine, in adult women at risk for recurrent UTI. Co-PI: Dr. Lindsay Nicolle and Dr. Godfrey Harding Project Description: As above Duration: Funding: MedImmune

Project Title: Rapid DNA based diagnostic tests Collaborator: Michael Mulvey Study description: As above Duration: 2003-2007 Funding: CRTI $2,999,426

Project Title: Reactivation of herpesviruses in transplant patients PI: Dr. Atul Humar (Toronto General Hospital); Co-PI: Dr. Graham Tipples; Study description: As above Duration: March – November 2004 Funding: Health Canada (GT); $15,000.

Project Title: Real-Time biosurveillance and response readiness using an interconnected, electronic information infrastructure: A region wide technology demonstration project at the Winnipeg Regional Health Authority Co-PI: Amin Kabani Project description: As above Duration: 2004 Funding: CRTI $1,900,000

Project Title: Real-time molecular surveillance of nosocomial pathogens PI: Michael Mulvey Study description: As above Duration: 2001 Funding: Office of the Chief Scientist; $135,000

Project Title: Recombinant urokinase in the treatment of occluded central venous catheters PI: Bruce Light Study Description: As above Duration: 2002 Funding: Abbott Labs; $25,000 Project Title: Regular routine chemoprophylaxis to reduce the incidence of sexually transmitted diseases and HIV infection among women who sell sex in Kenya PI: Stephen Moses Study description: This is a randomized, controlled trial examining the effect of regular, routine chemoprophylaxis (with monthly azithromycin) on reducing the incidence of STIs and HIV infection among female sex workers in Nairobi. Duration: July 1997 – June 2002 Funding: The Rockefeller Foundation; $588,000 USD

Project Title: Reliance Endoscope Cleaning Evaluation PI: Michelle Alfa Project description: As above Duration: 2004-2005 Funding: Steris :40,000 US

Project title: Resiquimod topical therapy of genital warts. PI: Fred Y. Aoki Collaborators: Greg Hammond, Pam Orr, Neil Simonsen Project Description: This is a placebo-controlled, double-blind study of topical Resiquimod for the treatment of genital warts. Duration: 2001 Funding: 3M Pharmaceuticals; $99,127

Project Title: Resistance with macrolides/ketolides PI: George Zhanel, Daryl Hoban, Dr S Douthwaite (Denmark) Study description: As above Duration: 2004-2005 Funding: Aventis $30,000

Project Title: Restoration after a CBRN attack PI: Dr. Merv Fingas, Environment Canada Collaborators: Kathryn Bernard; Stefan Wagener and others Project Description: This consortium is studying efficacy of methods and products used to decontaminate surfaces or sites should they be contaminated with biological, chemical or radionuclear weapons Duration: 2003-2006 onwards Funding: CRTI;  $2,000,000

Project Title: Restoration of Facilities and Areas after a CBRN Attack PI: Stefan Wagener Project Description: As above Duration: 2003-2005 Funding: CRTI $300,000

Project Title: Reverse genetics for Crimean-Congo Hemorrhagic fever virus. PI: Heinrich Feldmann Project description: The objective of this study is to develop and optimize a reverse genetics system (infectious cDNA clone) for the Crimean-Congo hemorrhagic fever (CCHF) virus, genus nairovirus, family Bunyaviridae. CCHF is a serious disease with high mortality (up to 60%) and the potential for human-to-human transmission. CCHF virus is classified as a Biosafety Level 4 (BSL-4) agent and is part of the Category A Pathogen List as defined by the Centers for Disease Control and Prevention (CDC). Case management and treatment would benefit from knowledge of the pathogenic mechanism, which is largely unknown. Using reverse genetics we would like to dissect the basic molecular mechanisms of the virus regarding promoter function, genome packaging, protein expression and protein-protein interactions. This will provide us with insight into the pathogenic mechanism of the virus and will help us to design concepts for antiviral treatment and vaccine development. During the next two years we will concentrate on the following aims: Aim #1: Determination of the L segment sequence of CCHF virus (strain IbAr10200) Aim #2: Optimization of CCHF virus-derived minigenome rescue Aim #3: Mutational promoter analysis of CCHF vRNA segments Aim #4: Rescue of recombinant infectious CCHF virus particles

The reverse genetics system (infectious clone) will be the key tool for our future studies. The system can be used to study the regulatory elements for viral transcription and replication. It can also be used to determine the functions of the viral proteins for the virus life cycle and to identify the functional domains of these proteins. Both studies will provide us with the necessary knowledge to develop antiviral drugs for treatment of CCHF. The system will also be helpful to study the immune response and the pathogenesis once appropriate animal models have been developed. Thus, the system will also be helpful to design appropriate vaccine candidates for CCHF. Vaccines and antivirals are two important defence strategies against weaponizable pathogens and could effectively be used to prevent the spread of an organism intentionally released in the environment. Thus, the development of the reverse genetics system is a crucial step on the way towards an effective defence system against this Category A listed pathogen. Duration: 2003-2004 Funding: National Institute of Allergy and Infectious Diseases (NIAID), Special Emphasis Panel; US $200,000

Project Title: The Role of a CD4 Polymorphism in Mother to Child HIV Transmission PI: Keith Fowke Project description: As above Duration: Jan 2003-Dec. 2004 Funding: Elizabeth Glaser Pediatric AIDS Foundation. . $155,000 per year.

Project Title: Role of dendritic cell in infection mediated inhibition of allergy PI: Xi Yang Study description: As above Duration: 2004-2009 Funding: CIHR; $735,000

Project Title: Roles of Hepatitis B and C virus proteins in immunity. PI: Jingxin Cao, Project Description: This project will adopt a genomic approach to study immunomodulatory functions of HBV and HCV proteins using a vaccinia expression system in a mouse model. Data generated from such studies will be extremely valuable in treating HBV and HCV infections and developing preventive/therapeutic vaccines for human use. Duration: 2002-2005 Funding: Health Canada Biotechnology Initiatives; $174,750

Project Title: The Role of Immune Activation and CD4 Gene Polymorphisms in an HIV Seroconversion Cohort from Kisumu, Kenya. PI: Keith Fowke Study description: As above Duration: April 2004-March 2005 Funding: CIHR; $83,000

Project Title: The role of soluble glycoproteins in the pathogenesis of Ebola and Marburg virus hemorrhagic fever. PI: Heinrich Feldmann Project description: Background: Marburg and Ebola viruses cause a fulminant hemorrhagic fever in humans and non-human primates. Following unspecific symptoms, patients display an increase in paraendothelial permeability, hypotension, coagulation disorders, and hemorrhages. Disturbances of the blood tissue barrier, primarily controlled by endothelial cells, and immunosuppression seem to be the pathogenic key factors of the disease. The endothelium is affected by two ways; directly by virus infection that leads to activation and lytic replication and indirectly by a mediator-induced inflammatory response. Those mediators originate from virus-activated cells of the mononuclear phagocytic system, especially macrophages, which are the primary target cells. The viral glycoproteins seem to be key factors for determining viral pathogenicity. Ebola viruses have developed specific strategies to express several products from their glycoprotein gene. The primary product is a small soluble glycoprotein precursor that subsequently is proteolytically processed into sGP and delta-peptide. RNA editing allows the expression of the transmembrane glycoprotein GP that mediates the spread of infection by binding to and fusion with target cells. Infectivity seems to be associated with proteolytic cleavage of GP into the subunits

GP1 and GP2 that are disulfide-linked and form the mature spike protein. Mature GP1,2 is quite unstable and GP1 is shed extensively during infection. Based on current data one can postulated that besides GP, which mediates virus entry, the different soluble glycosylated products (GP1, sGP, and delta-peptide) are important determinants in pathogenicity. Objective: Mononuclear phagocytotic and endothelial cells are the main target cells for filoviruses. An important factor in the pathogenesis seems to be target cell activation; however, the molecular events triggering activation are unknown. Since activation is an early event we hypothesize that it is mediated by viral glycoproteins. The proposal therefore focuses on the interaction of filoviral soluble glycoproteins (GP 1, sGP, delta-peptide) and the transmembrane glycoprotein (GP) with target cells and the identification of subsequent molecular events. Research Plan: The research plan contains of three parts. Firstly, expression, characterization and purification of the soluble glycoproteins of Ebola virus. The different glycoproteins will be expressed using eukaryotic expression systems (yeast, baculovirus). Prior to large-scale production and purification, the proper intracellular processing and transport of the proteins will be verified using transient expression systems. Secondly, development of assays that detect the soluble glycoproteins. The established assays will be used to determine the presence of the proteins in the blood of EBOV-infected monkeys and humans to confirm their importance in vivo. Thirdly, functional characterization of the soluble viral glycoproteins under flow conditions (rheometer). The recombinant expressed products will be used to activate endothelial cells and monocytes/macrophages in culture. The focus will be on adhesion molecule expression and increased permeability. Mesentery venules from infected monkeys will be investigated for evidence of vascular leakage. Recombinant vesicular stomatitis viruses expressing the soluble glycoproteins and finally virus mutants that will be generated using reverse genetics (infectious clone) will be used to demonstrate the effect of the proteins in animal models. All infectious work will be performed in a BSL4-laboratory. For our work we will establish a unique rheometer system which will allow us to perform the studies under physiological (flow) conditions. Expectations: The proposed project will give insight into the role of the different soluble glycoproteins in the pathogenesis of Ebola hemorrhagic fever. Understanding of the pathogenic mechanisms will allow to design therapeutic and preventative treatments. Duration: 2001-2004 Funding: Canadian Institutes of Health Research (CIHR); $ 484,077

Project Title: Safety and efficacy of platelet activating factor acetylhydrolase in patients with severe sepsis PI: Bruce Light Study description: As above Duration: 2002 Funding: ICOS Corp; $50,000

Project Title: Safety and efficacy of recombinant activated Protein C in patients with severe sepsis PI: Bruce Light Study Description: As above Duration: 2001 Funding: Lilly Pharmaceuticals; $61,000 Project Title: Safety and efficacy of tissue factor inhibitor in the treatment of patients with severe sepsis PI: Bruce Light Study description: As above Duration: 2001 Funding: Chiron Corp; $160,000

Project Title: SARS: A scientific collaboration to support public health response through vaccination. Co-PI: Michael Drebot and Timothy Booth Project description: As above Duration: 2003-2004 Funding: CIHR; $500,000

Study Title: Second and first trimester evaluation of risk of fetal trisomies. Co-participant: Paul Van Caeseele Study description: As above Duration: 2002-2006 Funding: CRT - $1,200,000; CIHR $655,833

Project Title: A seroprevalence study for West Nile virus: Organ transplant recipients Co-PI: Michael Drebot Project description: As above Duration: 2003-2004 Funding: CIHR; $30,000

Project Title: Sexual and perinatal transmission of HCV Co-PI: Chester Morris Study description: As above Duration: 1999-2001 Funding: Health Canada, $180,000

Project Title: Simulated Evaluator of Enzymatic Detergent PI: Michelle Alfa Project Description: Duration: 2004-2005 Funding: Case Medical Incorporated $9,000 (US)

Project Title: Single and multi-step mutational development with fluoroquinolones versus S. pneumoniae PI: George Zhanel, Daryl Hoban Study description: As above Duration: 2003 Funding: GlaxoSmithKline $56,000

Title of the project: SNP analysis and method standardization PI : Dr. Lai-King Ng Project description: The goal of this study is to produce a rapid detection tool for antimicrobial resistance in important pathogens and establish a proteomic approach to identify unique protein profiles (fingerprint) of bacterial pathogens using Neisseria gonorrhoeae as a model. We explored capillary electrophoresis (CE) as a high throughput and rapid method to detect point mutations contributing to antibiotic resistance in N. gonorrhoeae. Duration: Jan 2002 – Dec 2003 Funding : Office of the Chief Scientist, Health Canada; $35,000

Project Title: Strategic teaching and research grant for marginalized populations Co-PI: Chester Morris Study description: As above Duration: 2001-2003 Funding: $100,000.

Project title: Structure/function and assembly of the reovirus core, a model ribonucleoprotein complex PI: KM Coombs Project description: We are using defined assembly-defective temperature-sensitive (ts) mutants of mammalian reovirus (MRV) to delineate the assembly pathway of a model animal virus. During these studies we have also begun to address the molecular requirements of the viral RNA-dependant RNA polymerase (RdRp), a unique enzyme complex, which, because it is not present in host cells, may offer a logical target for chemotherapy directed against virtually all RNA viruses. The objectives of this continuing research program are fourfold; to complete the molecular characterization of our panel of MRV ts mutants, to characterize a newly-generated set of avian reovirus (ARV) ts mutants, to express relevant MRV and ARV proteins involved in particle assembly, and to map conformational differences in wild-type and ts mutant particles by limited proteolysis and mass spectrometry. Determining the detailed molecular mechanisms involved in virion replication and assembly, and in changes in structure/function of virion components during the replicative cycle, are crucial to understanding how to interrupt virion assembly and replication, thereby preventing spread of the virus. We are using genetic, biochemical, and biophysical analyses of intact native core structures and of a variety of assembly-defective mutant particles produced by each mutant at the non-permissive temperature. Defects in the viral replicative cycle and in virus assembly are being determined for remaining MRV mutants and for each of our newly- generated ARV mutants. Genes that harbour respective lesions are being determined by reassortant mapping and then sequenced to precisely locate the ts lesions. We will also clone and express relevant wild-type and mutant proteins, both to obtain purified proteins and to construct host cell lines that constitutively express complementing proteins, to assess the roles of each virus’ proteins in respective replication cycles. We also will exploit our capacity to use high-resolution mass spectrometry to map conformational changes present in the wild- type and mutant proteins to relate these alterations to the process of particle assembly and disassembly. Phase 1 Duration: Oct. 1998 – Sept 2003 Funding: MRC; $ 402,820 Phase 2 Duration: October 2003-September 2008 Funding: CIHR; $ 600,995

Project Title: Studies on the cell biology and biochemistry of chlamydiae host cell interactions PI: Grant McClarty Project Description: As above Duration: October 2001 – October 2006 Funding: CIHR; $610,000

Project Title: Study to Determine the Pharmacokinetics of antimycobacterial drugs in critically ill patients with tuberculosis. National Coordinator: R. Long, P. Orr (Manitoba Coordinator) Study description: As above Duration: Funding:

Project Title: A Study to Estimate the Protective Efficacy against Community Acquired Epidemic Influenza of Two Alternative Dosing Regimens of FluInsure (Intranasal) ID Biomedical Corporation. Principal Site Investigator: F. Aoki; Coinvestigator: P. Orr, G. Hammond, Study description: As above Duration: 2004 Funding:

Project Titles: Studies on Nipah virus. A. Pathogenesis and diagnostic tests development. B. Challenge study of a vaccine candidate for Nipah virus. PI: Dr. Hana Weingartl Co-PI: Dr. Markus Czub (Special Pathogens, NML. HC) Collaborative research: The work is done in collaboration with number of laboratories: HC Special pathogens, Australian Animal Health Laboratory (Dr. Deborah Middleton), Iowa State University (Dr. James Roth), University of Guelph (CIFIR) (Dr. Jeff Caswell), Merial (Dr. J.-C. Audonet), CDC and USDA. Dr. Weingartl lead the animal experimental part of the projects: challenge phase with the evaluation of the vaccine performance. Project Description: Nipah virus, emerged in Malaysia, is a zoonotic paramyxovirus causing fatal encephalitis in humans and porcine respiratory and encephalitic syndrome in pigs. Following September 2001, it became one of the potential bioterrorist agents. Any work with Nipah virus has to be done under BSL4 conditions. Dr. Weingartl lead the collaborative pathogenesis studies in pigs, which generated novel data on Nipah virus invasion of CNS (manuscript submitted for publication), and resulted in development of virus and antibody detection tests, and essential reagents for the rapid field test. Dr. Weingartl developed a pig model for vaccine testing, and lead the animal experimental part of the collaborative project testing the ALVAC based Nipah virus vaccine. Impact - international swine industry, human health, national defence; technology transfer - MTA for the rapid field test for detection of Nipah virus in swine to Chemicon Funding and duration: CRTI RD0196 (April 2002 – May 2006); CFIA TD W0207 (April 2002 – March 2005); NIH (R21 AI058038-01) – transfer part of the funds for the challenge experiment from Dr. James Roth; Total amount of award:475,000$ (technical salaries are covered by CFIA)

Project Title: Studies on SARS coronavirus (SARS-CoV). A. Susceptibility of chickens and pigs to SARS-CoV. B. Challenge study of a vaccine candidate for SARS. PI: Dr. Hana Weingartl Co-PI: Dr. Markus Czub (Special Pathogens, NML, HC), Dr. Jingxin Cao (Hepatits, NML, HC) Study description: An outbreak of severe acute respiratory syndrome (SARS) in humans, associated with a new coronavirus, was reported in South-East Asia, Europe and North America in early 2003. To address speculations that the virus originated in domesticated animals, basic scientific data on the livestock and poultry as a potential animal reservoir for the SARS coronavirus, was collected and later that year a team conducted one of the first vaccine trials in the world with a candidate vaccine developed at HC. Our results indicate that these animals do not play a role as an amplifying host for SARS coronavirus , and that liver complications may be related to vaccination and subsequent infection with SARS CoV. Funding: CFIA/HC operation budget

Study Title: Study of the type-specific antibody response to herpes simplex virus (HSV) using 2_D protein gel electrophoresis PI: Alberto Severini Collaborator: Michael Carpenter Study description: We used sera from HSV infected patients to identify antigen produced by HSV-1 and HSV-2, with the intent of identifying type-specific antigens that can be used for developing more specific serological assays or for finding markers of different clinical manifestation of herpes infection. We use 2-dimensional gel electrophoresis followed by Western blot and mass spectrometry to identify the antigens. Duration: 2002-2003 Funding: Health Canada Office of the Chief Scientist; $30,000

Project Title: Suppression of Radiation-Induced Germline Mutations in Mice by the Adaptive Response: PI: Douglas R. Boreham (PI,McMaster University), Collaborator: J David Knox Study description: There are a number of studies that show radiation can substantially increase germ-line mutation rate at mouse minisatellite loci. The high frequencies of spontaneous and induced mutations at minisatellite loci allow mutation induction to be measured at low doses of exposure in a small population, making minisatellite mutation a powerful tool to investigate radiation-induced heritable mutations. We have adopted this mutation assay to study the role of adaptive response in protecting mice against radiation-induced heritable defects. Our research is, to the best of our knowledge, the only research investigating the protective effect of the adaptive response in germline cells and heritable mutations. We have shown that male mice adapted to radiation with a low dose priming exposures do not pass on mutations to their offspring caused by subsequent large radiation exposures to the males. Duration: 2002-2004 Funding: U.S. Department of Energy, Low Dose Radiation Research Program-Basic Research: $76,530.

Project Title: Surrogate orthopox models to evaluate postexposure treatments for smallpox infection Co-PI: Heinrich Feldmann Project description: The primary objective of this project is to test VIG for protection efficacy by exploiting animal models of disease induced by two orthopox viruses that are closely related to vaccinia virus and variola virus (smallpox): rabbitpox (RPV) in rabbits and monkeypox (MPV) in nonhuman primates. The use of animal models to investigate poxvirus pathogenesis has recently been reviewed, and we have chosen two models that should provide important insights into the most appropriate therapeutic strategy to minimize disease pathogenesis for both generalized vaccinia complications and also possibly for monkeypox and smallpox in humans. Duration: 2004-2006 Funding: CRTI ; $ 1,700,000

Project Title: Survey evaluation of FDA policy change on re-use of single-use medical devices PI: Michelle Alfa Project Description Duration: 2001 Funding: Wayne State University, State Policy Center, $7,831 US

Project Title: Tamiflu study WV16193 PI: Fred Y Aoki Collaborators: Greg Hammond, Pam Orr, Neil Simonsen Project Description: This placebo-controlled study in multiple centres aimed to evaluate the effectiveness of famciclovir administered to family contacts of individuals with clinical influenza to assess the prophylactic efficacy of the drug in this setting. Duration: Winter 2000-2001 Funding : $8,432

Project Title: Testing effects of anti-viral agents PI: KM Coombs Project description: Viral diseases have historically been considered difficult to treat with selective antiviral chemotherapy. It was believed the viral replication cycle was so closely related to host cell metabolism that any attempt to suppress viral replication would kill or severely harm uninfected cells. Thus, few drugs are presently licensed for the treatment of viral infections and we still lack effective therapies for several important viral infections. These deficiencies highlight the need for further refinement of antiviral drug design and development. Mycophenolic acid (MPA) is a non-nucleoside non-competitive, reversible inhibitor of eukaryotic IMP dehydrogenase (IMPDH). IMPDH catalyzes the rate-limiting step in the de novo biosynthesis of purine mononucleotides and is involved in the early steps of GMP synthesis. IMPDH inhibitors are expected to mainly affect viral RNA and/or DNA synthesis when there is an increased need for synthesis as in the case of virus- infected cells. MPA is currently used clinically to prevent rejection of transplanted kidneys and hearts in combination with steroids and cyclosporine A, and had been reported inhibit replication of several viruses in vitro. In an effort to further delineate the antiviral mechanism and role of MPA, we examined the effects of MPA on mammalian (MRV) and avian (ARV) reovirus. MPA significantly inhibited MRV replication when used at concentrations less than those used as a clinical immunosuppressive agent. MRV inhibition was strain dependent and genetic reassortant mapping showed the M1 gene, which encodes the minor core protein mu2 that is thought to be a polymerase cofactor, is associated with the antiviral effects of MPA. Duration and Funding: This project, although currently not funded, has been underway since 2002

Project title: Therapeutic antibodies for Marburg and Ebola viruses Co-PI:: Heinrich Feldmann Project description: The filoviruses Ebola and Marburg are Category A Biological Agents as defined by Centres for Disease Control (CDC). These viruses cause an acute hemorrhagic fever in man with mortalities ranging from 23-90% and are prone to human-to-human transmission. Marburg virus was weaponized in the former USSR and a Japanese sect recently attempted to obtain Ebola virus for use in a bioterrorist attack. As neither therapeutic nor prophylactic treatments are available for these filoviruses they present a great risk to both military and civilian populations. Therapeutic antibodies have been suggested by many in the field to be the most promising strategy at present, as this approach provides a short-term strategy, something which is urgently required to address the CRTI priority of therapeutic measures for immediate reaction and near term consequence management capability. Passive protection against a lethal Ebola virus challenge has been demonstrated in small animals and post- exposure treatment with convalescent plasma seemed to have a protective effect in humans. A challenge in developing effective neutralizing antibodies has been the limited knowledge on viral proteins targeted by the host immune response. However, recent studies have developed a better understanding in this area and determined that the transmembrane glycoproteins (GP) are the key targets. Aspects of this work have been conducted at the Health Canada facilities in Winnipeg. Antibody therapy offers immediate protection to an infected individual and may be used in either protective or therapeutic applications. A large variety of antibody therapies including both polyclonal (pAbs) and monoclonal (mAbs) antibodies have been approved by regulatory agencies and are used in a wide range of disease indications in humans, including treatment of several viral infections Antibody based therapeutics are in fact the single largest class of biopharmaceuticals in clinical trials and are reported at various stages of development. In this project different forms of the viral transmembrane glycoproteins will be used as immunogens for the development of therapeutic monoclonal and polyclonal antibodies. The immunogens may also be used for the development of bispecific monoclonals (bsmAbs), a technology being developed at the University of Alberta in Dr. Suresh’s laboratory for unique ultrasensitive diagnostics and therapeutics. In a first step, caprine/ovine polyclonal antibodies will be raised and tested for efficacy and safety in Ebola and Marburg protection models (mice, guinea pigs). The caprine/ovine polyclonal antibody approach will provide a supply of therapeutic drug for short-term delivery. To provide a long-term solution, recombinant monoclonal antibodies will be developed for both Ebola and Marburg viruses. While the lead-time to generation of the recombinant antibodies will be longer than for the caprine/ovine antibodies, the recombinant products will be of better-defined specificity and will be available for indefinite supply. Mono-specific and bispecific antibodies to be generated in this study will be potential diagnostic reagents to be developed in future partnership or collaborations for use in surveillance and reconnaissance, which are objectives of DND. Antibody therapies for filoviruses will be developed that may be clinically and commercially viable. Such therapies have the attributes required to meet the CRTI's Investment Guidelines for Immediate Reaction and Near Term Consequence Management Capabilities. The availability of therapeutic remedies for use in Ebola and Marburg infection from human-to-human transmission addresses the CRTI priority area of Longer-term Consequence Management Issues not only addressing public confidence but also providing actual recovery possibilities preventing later epidemic spread. In addition the antibodies developed in this study may have dual use as diagnostic tools for rapid incident assessment. Duration: 2003-2004 Funding: CRTI; CDN $ 2,700,000

Project Title: Therapeutic Antibodies for Ebola and Marburg Viruses PI: Cangene Corporation; (Wendy Johnson) Collaborators: Health Canada, National Microbiology Laboratory, University of Alberta, CFIA Project Description: Ebola and Marburg viruses are among the deadliest known pathogens, causing severe hemorrhagic diseases with lethality rates of up to 83% for Marburg virus and 90% for Ebola Zaire virus. Since the discovery of filoviruses in 1967, outbreaks of Ebola Hemorrhagic Fever (EHF) and Marburg Hemorrhagic Fever (MHF) have been reported from several Central African countries, with the latest outbreaks reported in December 2001 in Gabon and Democratic Republic of the Congo. There is no question that the filoviruses, by virtue of their virulence and natural occurrence, represent a real threat for use as biological weapons. This is exemplified by their inclusion on the Centers for Disease Control and Prevention "Category A List" of pathogens. The former Soviet Union and Russia produced large quantities of Marburg and Ebola viruses until 1992. Soviet Union researchers quantified the aerosol infectivity of Marburg virus for monkeys, determining that no more than a few virions are required to cause infection. The Japanese terrorist cult Aum Shinrikyo unsuccessfully attempted to obtain Ebola virus as part of an effort to create biological weapons. In a bio-terrorism incident, rapid diagnostics and therapeutics will be the first line of defence, addressing the immediate health needs of the public and military. Long-term goals will include pre-exposure prophylaxis, which will be best achieved by vaccines. The goal of this project is to bring together Canadian federal, academic and industrial partners to develop therapeutic antibodies as a front-line, short-term defence against a bio-terrorist attack with either Ebola or Marburg viruses. Many workers in the field have suggested that therapeutic antibodies are the most promising post-exposure therapeutic strategy presently available. This approach provides short-term deliverables, which are urgently required to protect the public and first responders. The development of effective neutralizing antibodies to Marburg and Ebola viruses has been challenging because of the limited knowledge of the viral proteins that are important targets for the protective immune response. However, recent studies have contributed to a better understanding in this area and have identified the transmembrane glycoproteins (GP) as key targets. Aspects of this work have been conducted at the Health Canada facilities in Winnipeg. Passive protection against a lethal Ebola virus challenge has been demonstrated in small animals, and post-exposure treatment with convalescent plasma seemed to have a protective effect in humans. Furthermore, antibody therapy offers immediate protection to infected individuals and may be used in either preventive or therapeutic applications. A large variety of antibody therapies have been approved by regulatory agencies and are used in a wide range of disease indications in humans, including treatment of several viral infections. As a first step, large animal derived polyclonal antibodies will be raised and tested for efficacy and safety in Ebola and Marburg protection models. The large animal polyclonal antibody approach will provide a supply of therapeutic drugs for short-term delivery. To provide a longer-term solution, recombinant monoclonal antibodies will be developed for both Ebola and Marburg viruses. While the lead-time to generation of the recombinant antibodies will be longer than for the polyclonal antibodies, the recombinant products will be of better-defined specificity and will be available for indefinite supply. Researchers will evaluate soluble and membrane-bound versions of Ebola and Marburg virus glycoprotein (GP) antigens as candidates to raise monoclonal antibodies. All the recombinant proteins have already been successfully expressed and some of them are already available for immunization purposes. Currently researchers are characterizing other forms of recombinant glycoproteins and developing purification procedures for large-scale production. Two standard technologies will be employed to develop monoclonal antibody phage display technology (PDT) and hybridoma technology (HT). These two technologies are expected to produce two different and diverse sets of potential therapeutic drug candidates. The antibody candidates will be screened, and potentially high-affinity neutralizing antibodies will be chosen for full characterization. The HT-derived drug candidates will be evaluated in viral neutralization assays in vitro. Subsequently, they will be tested for their in vivo neutralization activity in rodent models. Promising candidates will be converted into mouse-human chimeric recombinant antibodies and expressed in Chinese Hamster Ovary (CHO) cells. These recombinant monoclonal antibodies will again be tested in an in vivo mouse model to confirm protective efficacy. The PDT-derived antibodies will be converted from phagemid-encoded clones to a soluble Fab (antibody fragment) format. These Fab fragments will be tested in an in vitro assay for viral neutralization. The neutralization characteristics of the Fab will be evaluated and, if required, the most promising Fab clones will be affinity matured by error-prone PCR. Selected clones will be converted into full-length human IgG antibodies and expressed as recombinant products in CHO cells. This project will result in the development of a panel of neutralizing antibodies for Ebola and Marburg viruses. The efficacy of antibodies will be screened initially by an in vitro assay of neutralization and then in rodent models of protection. It is projected that both PDT and HT will develop a minimum of 10 monoclonal antibodies. Each antibody will be purified to at least 95% purity using a laboratory scale GLP purification process. These fully human monoclonal antibodies will be tested in an in vivo mouse model to confirm protective efficacy, and further protection studies might be conducted. This program of work on monoclonal antibodies coupled with the short-term development of polyclonal antisera will provide federal and provincial authorities with the tools needed to protect first responders and the public from the threat of a biological terrorism attack with Marburg and Ebola viruses. Duration: 2003-2006 Funding: CRTI $2,626,300

Project Title: Therapeutic Benefit of Inducing a Stress Response in Scrapie Infected Mice. PI: J. David Knox Collaborators: Douglas R. Boreham (McMaster University), Stephanie Czub (Canadian Food Inspection Agency), Ron E. Mitchel (Atomic Energy of Canada Ltd.). Study description: Scrapie is a prion disease of sheep and goats with clinical symptoms that are characterized by a long latent period, progressive ataxia, tremor, wasting and death. The infectious agent is thought to be an aberrantly folded form of an endogenous cellular protein known as the prion protein (PrPC). Scrapie has been experimentally transmitted to laboratory rodents and these animals serve as experimental models for transmissible spongiform encephalopathies (TSEs) such as BSE (Bovine Spongiform Encephalopathy/Mad Cow Disease) and its human counterpart, Creutzfeldt Jacob disease (CJD). There has been increased urgency of late to study these diseases and develop treatments due to the recent outbreak in Britain of BSE in the cattle population and new variant CJD in people. BSE is suspected of causing the new variant CJD disease in humans in Britain. While BSE is considered a human pathogen Scrapie is not. There are no known cures or treatments for spongiform encephalopathies. The mechanism by which the misfolded form or the prion protein (PrPsc) leads to neurodegeneration is not known though several hypotheses have been proposed. i) the presence of a misfolded protein within the cell has a direct cytotoxic effect. ii) PrP plays a role in regulating the oxidative state of the cell and when converted to PrPsc it is no longer able to perform this function resulting in oxidative stress. iii) the formation of the characteristic PrPsc deposits result in the activation of the phagocytes of the brain, the microglia, that when unable to phagocytize the large PrPsc fibrils release the prodigious quantities of reactive oxygen species (ROS). These are released into the surrounding area resulting in injury of the adjacent cells. Enzymatic and nonenzymatic ROS scavenging systems, protein refolding chaperones, and DNA repair systems are all activated during the mammalian stress response. Our hypothesis is that by activating the endogenous systems designed to cope with all of the potentially injurious responses to TSE infection outlined above early in the course of the disease we will be able to slow or prevent the progression of the disease. The induction of these potent protective systems can be stimulated by a number of known exogenous signals including the mild fever-like conditions produced by whole body hyperthermia or by exposure to low doses of ionizing radiation (5-500 mGy total dose). Should these experiments demonstrate that the induction of a stress response can modulate the progression of prion diseases the potential is high that these treatments could quickly be advanced to clinical trials as there is currently no known cure or treatment for spongiform encephalopathies. Duration: 2003-2004 Funding: : Health Canada and Atomic Energy of Canada Ltd..

Project Title: Three centuries of tuberculosis among Western Canada’s First Nations: Spatio-temporal Variation in Epidemic Intensity and Transmission. PI: Dr. P. Hackett. Coinvestigators: Dr. K. Young, Dr. A. Herring, Dr. P. Orr. Study description: As above Duration: 2002- ongoing Funding: CIHR grant: $162,424.00

Project Title : Treatment of Acute Uncomplicated Pyelonephritis with Short Course Levofloxacin Principle Investigators: Dr. G. Zhanel, Dr. G.K.M. Harding, Dr. L.E. Nicolle Co-investigators: Dr. P. Nickerson, Dr. L. Bryski, Dr. J. Reda, Dr. D. Sitar Study description: This study is a pilot study of short course therapy with five days oral levofloxacin to develop preliminary information assessing the safety and effectiveness of this regimen for the treatment of pyelonephritis. Duration: Funding: Ortho-McNeil Pharmaceutical Inc; $335,758

Project Title: Treatment of Complicated Urinary Infection with Five Day High Dose Levofloxacin Principle Investigators: Dr. G. Zhanel, Dr. G.K.M. Harding, Dr. L.E. Nicolle Study description: This study will be a pilot study of the efficacy of 750 mg of levofloxacin once daily for 5 days in the treatment of complicated urinary tract infection. Funding: $183,156

Project Title: Treatment duration of complicated UTI PIs: George Zhanel, Lindsay Nicolle, Godfrey Harding, Peter Nickerson, Dan Sitar Study description: A study to assess the optimal duration of antibiotic therapy to treat complicated UTI Duration: 2003-2005 Funding: Ortho McNeill; $300,000

Project Title: Treatment duration of pyelonephritis PI: George Zhanel Collaborators: Lindsay Nicolle, Godfrey Harding, P Nickerson, Dan Sitar Study description: A study to assess the optimal duration of treatment of pyelonephritis Duration: 2003-2005 Funding: Ortho McNeill $452,000

Project Title: Trends in hepatitis testing and Infection in Manitoba PI: MR Dawood Project description: We studied the trends in hepatitis viruses (Hep. A, B and C) infection in Manitoba from 1992 to 2003. The data was based on actual testing of clinical samples submitted for hepatitis testing. An Access database was developed to include all patients’ demographic data, and test results. The trends of different hepatitis virus infection and co-infection were analyzed to determine different groups at high risk to certain viral infection. The generated data are important for different programs planning and public health services delivery. Duration: 1992-2003 Funding: Health Canada; $60,000

Project Title: Tuberculosis Epidemiology Surveillance Consortium Collaborator: Amin Kabani Project description: As above Duration: 2001 Funding: Center for Disease Control, Atlanta.

Project Title: Use of Diagnostic Tests For Lower Respiratory Tract Infection in Long Term Care Facilities Study Investigators: Dr. L.E. Nicolle, Dr. G. Mazowita, Dr. M. Loeb, Dr. A. Simor, Dr. B. Liu, Dr. S. McNeil Study description: This study is being undertaken in several Canadian sites to describe current use of diagnostic tests for potential lower respiratory tract infections in residents of long term care facilities. In addition, it will examine some of the associations which determine the frequency of use of diagnostic tests. The specific study objectives include: a) To describe the current use and variability of use of specified diagnostic tests in managing potential episodes of lower respiratory infection in long term care facilities in Canada. b) To characterize associations of a high or low frequency of diagnostic test use including institutional characteristics, patient characteristics and presentations of the acute episode. c) To describe the frequency with which diagnostic test results, particularly sputum cultures lead to alteration and management. d) To correlate patient outcome with use of a specific test or combinations of tests for an episode of presumed lower respiratory tract infection. Duration: January 2003 – December 2003 Funding: Canadian Institutes for Health Research, Interdisciplinary Health Research Grant; $231,400

Project Title: Use of DNA microarrays for varicella-zoster virus strain analysis PI: Dr. Graham Tipples; Study description: As above Duration: April 2002 – March 2005 Funding: Health Canada Biotechnology Initiative; $126,000.

Project Title: The use of novel recombinant DNA/protein reagents for the development of improved diagnostic tests for viral diseases CoPI: Michael Drebot Project Description: As above Duration: 1999-2001 Funding: Health Canada Genomics R&D Biotechnology Grants; $158,583/year

Project Title: Use of reovirus as a “biomarker” for medical device disinfection PI: M Alfa; coPI: KM Coombs Project description: There is a growing appreciation that infections in the hospital setting can be spread through ineffective cleaning of re-useable medical devices. Some such devices (ie. endoscopes) are fragile, consist of numerous long thin hollow components, and may not be conducive to “deep sterilization” (for example, autoclaving), so may need be chemically sterilized, which raises the possibility that chemical may not access all parts of these instruments. Another variable that is currently poorly understood is what protective effect, if any, material deposition and drying may have on protecting pathogenic organisms within such instruments from the disinfecting effects of chemicals. Dr. M Alfa has been studying the effects of chemical disinfection of such instruments on survival of model bacterial organisms. We have initiated a collaborative project to extend these studies to examine effects of chemical disinfectants and drying on survival of viruses, including our model “biomarker”, the non-enveloped mammalian reovirus as well as an enveloped virus (Sindbis virus, a model for Togaviruses, which includes Rubella virus, West Nile virus and various Equine Encephalitis viruses). Initial results indicate that the non-enveloped reovirus can survive long periods of time in a dried state and that it is sensitive to chemical disinfectants such as glutaraldehyde and hydrogen peroxide, but less so than many indicator bacteria. The enveloped Sindbis virus is significantly more sensitive to drying and our future plans are to examine sensitivity of Sindbis to various chemical disinfectants. Duration and Funding: This project, although currently not funded, has been underway since 2003

Project Title: Use of reovirus as a “biomarker” for wastewater treatment PI: KM Coombs; Collaborators: JA Oleszkiewicz Project description: The development of adequate pathogen indicators is one of the key priorities for the wastewater management industry, municipalities, international water and wastewater organizations, Water Environment Research Foundation (WERF) and the United States Environmental Protection Agency (U.S. EPA). Recent events (ex. Walkerton, ON and North Battleford, SK in Canada) have demonstrated the importance, and weakness, of municipal water treatment and water safety. Indicator organisms are monitored to provide information regarding potential microbial pathogens in a waste matrix. Currently, the process of spiking waste matrices with non-indigenous surrogate pathogens is labor-intensive, time consuming and potentially dangerous to human health. For example, poliovirus has been used in some cases as a viral indicator of water purity, but, with its imminent Biosafety Level upgrading, alternative viruses will be required. Indicator organisms are nonpathogenic, indigenous to sludge, wastewater or run-off, less expensive to assess and rapidly measured. Indicator organisms are needed to assess the presence of pathogens in the waste matrices, to monitor the fate of potential pathogens during processing and land application and to provide a risk assessment for human health based on the presence of these indicators. Therefore, it is clear that development of appropriate indicators and surrogate organisms is essential for accurate assessment of pathogen concentration and assessing their risk to human health. We have started collaborative studies with Dr. JA Oleszkiewicz, an environmental engineer at the University of Manitoba to use reovirus as a “biomarker” indicator organism to test the efficacy of various wastewater treatment regimens. Results from this project, which examined effects of treating virus with lime, have been reported at a variety of scientific conferences. We are currently extending these studies by examining the effects of other treatments, including ultraviolet irradiation, on virus survival. Duration and Funding: This project, although currently not funded, has been underway since 1999 and is the subject of a pending application to the Water Environment Research Foundation

Project Title: Using photo-elicitation as a technique to document sex workers lives in Kenya PI: Chester Morris Study description: As above Duration: 2004-2005 Funding: Rockefeller Foundation; $67,000

Project Title: Vasopressin in the treatment of Septic Shock PI: Dr J Russel of UBC Site Investigator: Bruce Light Study Description: As above Duration: 2002-2003 Funding: CIHR; $79,040

Project Title: Viral Genomics. Diagnostic and expression DNA micro-arrays for influenza, encephalitis associated viruses and herpes simplex virus CoPI: Michael Drebot Project description: As above Duration: 2001-2004 Funding: Health Canada Genomics Biotechnology Grant; 150,000/year

Project Title: WHO TB Specimen Bank Clinical Enrollment and Collection Study Collaborator: Amin Kabani Project description: As above Duration: 2001 Funding: Health Canada

Project Title: Whole genome sequencing of varicella-zoster virus strains PI: Dr. Graham Tipples; Co-PI: Dr. Charles Grose (University of Iowa) and Shaun Tyler (NML); Study description: As above Duration: September 2002 – April 2003 Funding: Office of the Chief Scientist, Health Canada; $40,000.

Project title: Winnipeg injection drug user study: integrating social network and genetic data for program planning and delivery PI: John Whylie Co-investigator: MR Dawood Project description: We studied the prevalence of HCV among injection drug users in Manitoba and their contacts. The HCV RNA was amplified from infected individuals and their contacts. Nucleic acid sequence was determined for different isolates. And sequence analysis was performed to determine the correlation between isolates from different patients and their contacts. Duration: Funding: Health Canada; $41,340

Project Title: Workplace interventions in Malawi PI: Chester Morris Study description: As above Duration: 1999-2001 Funding: US Department of Labor; $60,000

DEVELOPMENT PROJECTS

Project Title: REGIONAL AIDS TRAINING NETWORK PI: Frank Plummer– Phase 1 (March 1997-April 2002); Stephen Moses – Phase 2 (May 2002-April 2007) U of Manitoba Co-Director: (March 1997 – October 2003) L. Gelmon U of Nairobi Co-Director (March 1997 – October 2003) J. Bwayo Project Description: This project has created an independent African NGO, whose purpose is the establishment of a network of training institutions in Eastern and Southern Africa which are delivering a series of short course in AIDS/STI related disciplines, aimed at trainers and programme managers. In its first seven years, RATN through a network of more than twenty partner institutions, delivered more than 200 training courses to more than 2000 trainees from 24 African countries. Duration: 1997 – onwards Funding: multiple sources - CIDA, World Bank, SIDA, DCI, UNAIDS, UNICEF, Action Aid, FHI, HRSA, Univ Washington, Pop Council, Total amount – Phase 1  CAD $8,000,000 ($5,000,000 from CIDA) - Phase 2  CAD $10,000,000($4,610,523 from CIDA)

Project Title: AIDS PREVENTION AND CONTROL IN INDIA PI: Stephen Moses Project description: A consortium led by the University of Manitoba is the Canadian Executing Agency (CEA) to design and deliver a bilateral HIV/AIDS prevention and control project in India. Duration: November 1999 – March 2006 Funding: CIDA $13,645,476

Project Title: STRENGTHENING HIV/STI PROGRAM IN KENYA FOR VULNERABLE GROUPS PI: Frank Plummer (Phases I,II); Stephen Moses (Phase III) Co-Directors: Chester Morris (Phase III), Elizabeth Ngugi (Phases I,II,III) Project description: Phase III: The purpose of this project is to catalyze the widespread implementation of evidence-based STD/AIDS prevention and care programming in Kenya. The project works with government, the donor community, non-governmental organizations, community-based organizations and the private sector to scale up targeted STD/AIDS prevention programs for vulnerable groups of women and men, as well as enhanced STD management services. Duration: 1993-2007 Funding: CIDA, Phase III $7,800,000 Project Title: SCALING UP HIV PREVENTION IN KARNATAKA. PI: James Blanchard; Co-Investigator: Stephen Moses Project Description: The principal goal of this project is to reduce the transmission of HIV and STIs in the Indian state of Karnataka. Duration: December 2003 – November 2008 Funding: Bill & Melinda Gates Foundation; $17,000,000 USD