Genes for three maize homologs (CenpcA, CenpcB, and CenpcC) of the conserved kinetochore assembly protein known as centromere protein C (CENPC) have been identified. The C-terminal portion of maize CENPC shares similarity with mammalian CENPC and its yeast homolog Mif2p over a 23-amino acid region known as region I. Immunolocalization experiments combined with three-dimensional light microscopy demonstrated that CENPC is a component of the kinetochore throughout interphase, mitosis, and meiosis. It is shown that sister kinetochore separation occurs in two discrete phases during meiosis. A partial separation of sister kinetochores occurs in prometaphase I, and a complete separation occurs in prometaphase II. CENPC is absent on structures known as neocentromeres that, in maize, demonstrate poleward movement but lack other important features of centromeres/kinetochores. CENPC and a previously identified centromeric DNA sequence interact closely but do not strictly colocalize on meiotic chromosomes. These and other data indicate that CENPC occupies an inner domain of the maize kinetochore.
A blocking ELISA targeting an immunodominant West Nile epitope on the West Nile Virus NSI protein was assessed for the detection of West Nile-specific antibodies in blood samples collected from 584 sentinel chickens and 238 wild birds collected in New Jersey from May-December 2000. Ten mallard ducks (Aftus plat!lrlry~fchos) experimentally infected with West Nile virus and six uninfected controls were also tested. The ELISA proved specific in detecting WNV antibodies in 9/10 chickens and 414 wild birds previously confirmed as positive by Plaque Reduction Neutralization test (PRNT) at the Center for Disease Control, Division of Vector Borne Diseases, Fort Collins, CO, USA (CDC). Nine out of the ten experimentally infected mallard ducks also tested positive for WN antibodies in the blocking ELISA, while 616 uninfected controls did not. Additionally, 1705 wild birds, collected in New Jersey from December 2000-November 2001 and Long Island, New York between November 1999 and August 2001 were also tested for WN antibodies by the blocking ELISA. These tests identified 30 positive specimens, 12 of which had formalin-fixed tissues available to allow detection of WN specific viral antigen in various tissues by WNV-specific immunohistochemistry. Our results indicate that rapid and specific detection of antibodies to WN virus in sera from a range of avian species by blocking ELISA is an effective strategy for WN Virus surveillance in avian hosts. In combination with detection of WN-specific antigens in tissues by immunohistochemistry (IHC) the blocking ELISA will also be useful for confirming WN infection in diseased birds.
The American crow (Corvus brachyrhynchos) is known to suffer 100% mortality from infection with the New York 1999 strain of West Nile virus (WNV). Following the initial detection of WNV in North America in 1999, we measured prevalence of WNV-reactive antibodies (''seroprevalence'') in free-ranging American and fish crows (Corvus ossifragus) of central New Jersey after each transmission season through 2005. In 2002, seroprevalence in American crow juveniles increased to 14% from the 5% of the previous year, potentially indicating increased survival in this species. Using the annual seroprevalence measurements and the number of human West Nile neuroinvasive disease cases as a surrogate for WNV transmission intensity, we developed a model to estimate the annual WNV-associated mortality rates among both of these crow species. Our model supports the hypothesis that mortality is changing over time; the WNV-associated mortality rate declined over time by 1.5% for American crow and by 1.1% for fish crow. The probability that the trend in mortality was negative was 90% for the American crow and 60% for the fish crow.
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