Characterization and autoproteolytic activity of N[superscript pro] of bovine viral diarrhea virus

http://www.worldcat.org/oclc/4155684

DNA vaccination and recombinant protein expression to identify immunogenic and protective genes from M avium subsp paratuberculosis

Paratuberculosis (Johne\u27s disease) is a chronic granulomatous infection of cattle, caused by the intracellular bacterium Mycobacterium avium subsp. paratuberculosis (referred to hereafter as M. paratuberculosis). Currently available vaccines are not effective at preventing infection or transmission of the bacterium to other animals. The objectives of this dissertation were to develop and test DNA vaccines that prevent M. paratuberculosis infection, to examine the host immune response induced by protective DNA vaccines, and to identify immunogenic proteins from M. paratuberculosis. In an effort to identify protective M. paratuberculosis sequences, a genomic DNA expression library was generated and subdivided into pools of clones. Eleven clone pools were evaluated to determine DNA vaccine efficacy by immunizing mice via gene gun delivery and challenging with live, virulent M. paratuberculosis. Four clone pools demonstrated a significant reduction (P \u3c 0.05) of M. paratuberculosis infection in mice when compared to other clone pools and non-vaccinated, infected control mice. In a second study, one of the protective clone pools was partitioned into 10 new clone arrays of 108 clones each and an immunization experiment was performed to identify protective clone arrays. When groups of mice were immunized with clone array DNA via gene gun, four clone arrays provided significant protection (P \u3c 0.05) from M. paratuberculosis challenge. Comparison of the nucleotide sequences from protective clone arrays identified 26 sequences that may have contributed to the protection observed in these mice. Additionally, protective clone arrays were found to induce IL-10 and IL-12 cytokine production, which indicated induction of both TH1 and TH2 immune responses. In a final experiment, the efficacy of the M. paratuberculosis 19 kDa lipoprotein was evaluated as an immunomodulator in cattle with Johne\u27s disease. Control, non-infected cattle did not produce antibodies or cell-mediated immune responses against the 19 kDa protein. However, the 19 kDa protein induced cellular immune responses (IFN-gamma production) in subclinically infected cattle and induced humoral immune responses (19 kDa-specific antibody production) in clinically infected cattle. Taken together, these three studies have identified a number of M. paratuberculosis antigens that may aid in the development of vaccines to prevent Johne\u27s disease

Immunoreactivity of the Mycobacterium avium subsp. paratuberculosis 19-kDa lipoprotein

BACKGROUND: The Mycobacterium tuberculosis 19-kDa lipoprotein has been reported to stimulate both T and B cell responses as well as induce a number of Th1 cytokines. In order to evaluate the Mycobacterium avium subsp. paratuberculosis (M. avium subsp. paratuberculosis) 19-kDa lipoprotein as an immunomodulator in cattle with Johne's disease, the gene encoding the 19-kDa protein (MAP0261c) was analyzed. RESULTS: MAP0261c is conserved in mycobacteria, showing a 95% amino acid identity in M. avium subspecies avium, 84% in M. intracellulare and 76% in M. bovis and M. tuberculosis. MAP0261c was cloned, expressed, and purified as a fusion protein with the maltose-binding protein (MBP-19 kDa) in Escherichia coli. IFN-γ production was measured from 21 naturally infected and 9 control cattle after peripheral blood mononuclear cells (PBMCs) were stimulated with a whole cell lysate (WCL) of M. avium subsp. paratuberculosis or the recombinant MBP-19 kDa. Overall, the mean response to MBP-19 kDa was not as strong as the mean response to the WCL. By comparison, cells from control, non-infected cattle did not produce IFN-γ after stimulation with either WCL or MBP-19 kDa. To assess the humoral immune response to the 19-kDa protein, sera from cattle with clinical Johne's disease were used in immunoblot analysis. Reactivity to MBP-19 kDa protein, but not MBP alone, was observed in 9 of 14 infected cattle. Antibodies to the 19-kDa protein were not observed in 8 of 9 control cows. CONCLUSIONS: Collectively, these results demonstrate that while the 19-kDa protein from M. avium subsp. paratuberculosis stimulates a humoral immune response and weak IFN-γ production in infected cattle, the elicited responses are not strong enough to be used in a sensitive diagnostic assay

Use of intra-medullary stacked nailing in the reduction of proximal plastic deformity in a pediatric Monteggia fracture:a case report

<p>Abstract</p> <p>Introduction</p> <p>In a Monteggia fracture dislocation, it is important to reduce the ulnar fracture completely. Extensive plastic deformation of the proximal ulna may make reduction by closed manipulation impossible.</p> <p>Case presentation</p> <p>We report the case of a four-year-old Caucasian boy in whom the plastic deformation of the proximal ulna was reduced, and this reduction was maintained, using intra-medullary stacked nailing.</p> <p>Conclusion</p> <p>The technique of stacked nailing is a useful addition to the armamentarium in the management of the potentially awkward Monteggia fracture.</p

Ticks and Tularemia: Do We Know What We Don't Know?

Francisella tularensis, the causative agent of the zoonotic disease tularemia, is characterized by high morbidity and mortality rates in over 190 different mammalian species, including humans. Based on its low infectious dose, multiple routes of infection, and ability to induce rapid and lethal disease, F. tularensis has been recognized as a severe public health threat—being designated as a NIH Category A Priority Pathogen and a CDC Tier 1 Select Agent. Despite concerns over its use as a bioweapon, most U.S. tularemia cases are tick-mediated and ticks are believed to be the major environmental reservoir for F. tularensis in the U.S. The American dog tick (Dermacentor variabilis) has been reported to be the primary tick vector for F. tularensis, but the lone star tick (Amblyomma americanum) and other tick species also have been shown to harbor F. tularensis. This review highlights what is known, not known, and is debated, about the roles of different tick species as environmental reservoirs and transmission vectors for a variety of F. tularensis genotypes/strains

Revisional Surgery for Hallux Valgus with Serial Osteotomies at Two Levels

The aetiology and form of hallux valgus (HV) is varied with many corrective procedures described. We report a 39-year-old woman, previously treated with a Chevron osteotomy, who presented with recurrent right HV, metatarsus primus varus, and associated bunion. Osteotomies were performed at two levels as a revisional procedure. This report highlights (1) limitations of the Chevron osteotomy and (2) the revisional procedure of the two level osteotomies: (i) proximal opening-wedge basal osteotomy and (ii) distal short Scarf with medial closing wedges. If a Chevron osteotomy is used inappropriately, for example, in an attempt to correct too large a deformity, it may angulate laterally causing a malunion with an increased distal metatarsal articular angle. Secondly, it is feasible to correct this combined deformity using a combination of proximal opening-wedge and distal short Scarf osteotomies

Tick Extracellular Vesicles Enable Arthropod Feeding and Promote Distinct Outcomes of Bacterial Infection

Extracellular vesicles are thought to facilitate pathogen transmission from arthropods to humans and other animals. Here, we reveal that pathogen spreading from arthropods to the mammalian host is multifaceted. Extracellular vesicles from Ixodes scapularis enable tick feeding and promote infection of the mildly virulent rickettsial agent Anaplasma phagocytophilum through the SNARE proteins Vamp33 and Synaptobrevin 2 and dendritic epidermal T cells. However, extracellular vesicles from the tick Dermacentor andersoni mitigate microbial spreading caused by the lethal pathogen Francisella tularensis. Collectively, we establish that tick extracellular vesicles foster distinct outcomes of bacterial infection and assist in vector feeding by acting on skin immunity. Thus, the biology of arthropods should be taken into consideration when developing strategies to control vector-borne diseases

The Contribution of Occult Precipitation to Nutrient Deposition on the West Coast of South Africa

The Strandveld mediterranean-ecosystem of the west coast of South Africa supports floristically diverse vegetation growing on mostly nutrient-poor aeolian sands and extending from the Atlantic Ocean tens of kilometers inland. The cold Benguela current upwelling interacts with warm onshore southerly winds in summer causing coastal fogs in this region. We hypothesized that fog and other forms of occult precipitation contribute moisture and nutrients to the vegetation. We measured occult precipitation over one year along a transect running inland in the direction of the prevailing wind and compared the nutrient concentrations with those in rainwater. Occult deposition rates of P, N, K, Mg, Ca, Na, Al and Fe all decreased with distance from the ocean. Furthermore, ratios of cations to Na were similar to those of seawater, suggesting a marine origin for these. In contrast, N and P ratios in occult precipitation were higher than in seawater. We speculate that this is due to marine foam contributing to occult precipitation. Nutrient loss in leaf litter from dominant shrub species was measured to indicate nutrient demand. We estimated that occult precipitation could meet the demand of the dominant shrubby species for annual N, P, K and Ca. Of these species, those with small leaves intercepted more moisture and nutrients than those with larger leaves and could take up foliar deposits of glycine, NO3-, NH4 + and Li (as tracer for K) through leaf surfaces. We conclude that occult deposition together with rainfall deposition are potentially important nutrient and moisture sources for the Strandveld vegetation that contribute to this vegetation being floristically distinct from neighbouring nutrient-poor Fynbos vegetation

Pathways between Primary Production and Fisheries Yields of Large Marine Ecosystems

The shift in marine resource management from a compartmentalized approach of dealing with resources on a species basis to an approach based on management of spatially defined ecosystems requires an accurate accounting of energy flow. The flow of energy from primary production through the food web will ultimately limit upper trophic-level fishery yields. In this work, we examine the relationship between yield and several metrics including net primary production, chlorophyll concentration, particle-export ratio, and the ratio of secondary to primary production. We also evaluate the relationship between yield and two additional rate measures that describe the export of energy from the pelagic food web, particle export flux and mesozooplankton productivity. We found primary production is a poor predictor of global fishery yields for a sample of 52 large marine ecosystems. However, chlorophyll concentration, particle-export ratio, and the ratio of secondary to primary production were positively associated with yields. The latter two measures provide greater mechanistic insight into factors controlling fishery production than chlorophyll concentration alone. Particle export flux and mesozooplankton productivity were also significantly related to yield on a global basis. Collectively, our analyses suggest that factors related to the export of energy from pelagic food webs are critical to defining patterns of fishery yields. Such trophic patterns are associated with temperature and latitude and hence greater yields are associated with colder, high latitude ecosystems

The bii4africa dataset of faunal and floral population intactness estimates across Africa’s major land uses

Sub-Saharan Africa is under-represented in global biodiversity datasets, particularly regarding the impact of land use on species’ population abundances. Drawing on recent advances in expert elicitation to ensure data consistency, 200 experts were convened using a modified-Delphi process to estimate ‘intactness scores’: the remaining proportion of an ‘intact’ reference population of a species group in a particular land use, on a scale from 0 (no remaining individuals) to 1 (same abundance as the reference) and, in rare cases, to 2 (populations that thrive in human-modified landscapes). The resulting bii4africa dataset contains intactness scores representing terrestrial vertebrates (tetrapods: ±5,400 amphibians, reptiles, birds, mammals) and vascular plants (±45,000 forbs, graminoids, trees, shrubs) in sub-Saharan Africa across the region’s major land uses (urban, cropland, rangeland, plantation, protected, etc.) and intensities (e.g., large-scale vs smallholder cropland). This dataset was co-produced as part of the Biodiversity Intactness Index for Africa Project. Additional uses include assessing ecosystem condition; rectifying geographic/ taxonomic biases in global biodiversity indicators and maps; and informing the Red List of Ecosystems