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Descriptions of Pilof Projects Project I: The study of prevalence, genotype distribution, and molecular variability of isolates of hepatitis C virus in the Asian part of Russia Description: This project focuses on sequencing and identifying genotypic variants of hepatitis C virus (HCV) in the Asian Russian population to determine the extent of variability of the virus in the region. Importance: HCV is a serious public health problem in Russia, the United States, and globally. Approximately 2 percent of blood donors in Russia are infected with HCV, resulting in a 20 to 40 percent prevalence of infection in recipients of multiple transfusions. Fifteen isolates of HCV have been partially sequenced to date. This has led to the discovery of several nontypical genotypes that appear to have evolved independently in the isolated populations of Siberia and the Far East. There is a need for additional data on the extent of variability of the virus in Asian Russia (and elsewhere) for three reasons: (~) to determine whether commercially available tests can detect all current genotypic variants of HCV, (2) to ascertain how well vaccines in development will protect against these variants, and (3) and to provide an additional means for estimating the prevalence of HCV infection in the general population. The findings of this project should usefi~ly inform national HCV prevention and control programs. Project 2: Monkeypox virus genome Description: This project focuses on sequencing the monkeypox virus genome. Importance: Monkeypox is a classic emerging infectious disease. Sequencing the monkeypox virus genome will facilitate development of species-specific diagnostics based on polymerase chain reaction. In addition, comparison of the monkeypox virus genome with that of variola (smallpox) mav reveal substantial dunlicatic)n of gene filnctinn.~ thereLv rontribiltina e~nti~1 ' ~ ~ ~ ~ -rid - ~ ~ _ ~ __. ^,~ __~_^,~,~, information relevant to the planned worldwide destruction of variola in 1999. Better understanding of the relation of structure to function in the monkeypod virus genome is also expected to provide insight into the rational design of effective antiviral drugs and therapeutic strategies for monkeypod and other orthopox viruses. Project 3: Russia Description: This project is cataloguing the genetic and serologic variability of hantaviruses collected from the Asian part of Russia. Importance: Hemorrhagic fever with renal syndrome is a significant cause of human morbidity and mortality in the Asian part of Russia, with the disease extending to South Korea and China. Strains of classic Hantaan virus, found in China and Korea, are known to occur in far eastern Russia, whereas Puumala virus, predominant in European Russia, extends into Siberia and the Far East. in addition, other newly recognized Hantaviruses such as Khabarovsk virus exist in eastern Russia, but their potential to cause human illness has yet to be determined. Hantaviruses are emerging throughout the world, and it is currently unknown where new Asian strains fit in the phylogenetic tree. This study should yield important information about the serologic and genetic variability of Hantaviruses and help identify rodent hosts and risk factors for this important group of viral pathogens. These findings, in turn, could help inform research and development in support of effective vaccines against Hantavirus infection in the United States and Russia. Project 4: Description: Study of the genetic and serologic diversity of Hantavirus in the Asian part of Development of advanced diagnostic kit for opisthorchiasis in human patients This project is developing an advanced diagnostic kit for human opisthorchiasis. Importance: The parasitic liver fluke Opisthorchis felineus represents a significant human health problem in much of Russia, with an estimated 10 to 20 million human infections in Siberia alone. Liver flukes in the same family are also found in contaminated fish in the northwestern
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United States. Current diagnostic procedures rely on direct stool examination, which is time- consuming, technically difficult, and expensive. An enzyme immunoassay has been developed for serologic diagnosis but is limited in value because it is not able to differentiate between current infections and cured individuals. An effective treatment with phenolics is available, but these drugs are too toxic to use except to treat active infection; thus, there is an urgent need for an improved diagnostic test to differentiate active cases rapidly, accurately, and with minimum cost. Sufficient human samples are readily available to ensure that substantive evaluation of candidate assays will be conducted promptly, with preliminary results likely to be available during 1997. Projects: Molecular biological and immunochemical analysis of clinical strains of tuberculosis and mycobacteriosis Description: This project focuses on characterization of different strains of mycobacteria in Russian patients diagnosed with tuberculosis (TB). Importance: TB, particularly drug-resistant TB, represents a serious threat to the United States and is increasing in Russia in epidemic proportions. This project is characterizing different strains of mycobacteria isolated from Russian patients diagnosed with TB and is determining the spectrum of drug resistance among them. The relation between strain virulence and the spectrum and degree of drug resistance will be explored by identifying the genes responsible for drug resistance. New antibiotics under development in Russia will be tested for their potency against these clinical strains. This project will heln strengthen Russian can~hililv in nr1~1re~ina the emerging TB epidemic in Russia. - -r ~--~-----~ ~ ~~~~-~~~ ~-r ~~ ~-~~J ~~~ ~~~~ ~~~~~~= ~~~~ Project6: Investigation of the immunological effectivity of delivery in vivo of the Brucella main outer membrane protein by anthrax toxin components Description: This project is an initial step toward the eventual goal of producing an effective recombinant protein vaccine or vaccine mixture for veterinary use against Brucelia abortus and for protection of occupationally exposed personnel. Importance: Human brucellosis is a disease caused by species of the bacterium Brucelia. In humans it is seriously debilitating but seldom lethal. Its principal reservoirs are cattle, sheep, and swine. Human exposure is principally from direct contact with infected animals and animal products, including consumption of unpasteurized milk and milk products from infected animals. Control of the disease in humans occurs mainly by avoiding the consumption of unpasteurized milk or milk products and contact with infected animals, sacrificing infected animals and herds, and, in areas where brucellosis is endemic, veterinary vaccination. None of the current vaccines against brucellosis is completely satisfactory; their shortcomings include incomplete protection, induction of abortion, and occasional infectivity to humans. This protocol calls for the construction of chimeric genes expressing anthrax lethal factor (LF)-Brucella outer membrane protein (OMP) fusion proteins and testing of the resulting chimeric proteins when administered together with anthrax protective factor for immunological effectiveness against Brucella abortus. The I,F- protective antigen (PA) cell delivery system holds great promise for an improved brucellosis vaccine, in particular, and for more effective disease prevention in the United States and Russia generally.