Activation of apoptosis in vivo by a hydrocarbon-stapled BH3 helix

BCL-2 family proteins constitute a critical control point for the regulation of apoptosis. Protein interaction between BCL-2 members is a prominent mechanism of control and is mediated through the amphipathic alpha-helical BH3 segment, an essential death domain. We used a chemical strategy, termed hydrocarbon stapling, to generate BH3 peptides with improved pharmacologic properties. The stapled peptides, called "stabilized alpha-helix of BCL-2 domains" (SAHBs), proved to be helical, protease-resistant, and cell-permeable molecules that bound with increased affinity to multidomain BCL-2 member pockets. A SAHB of the BH3 domain from the BID protein specifically activated the apoptotic pathway to kill leukemia cells. In addition, SAHB effectively inhibited the growth of human leukemia xenografts in vivo. Hydrocarbon stapling of native peptides may provide a useful strategy for experimental and therapeutic modulation of protein-protein interactions in many signaling pathways.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

Evaluation of surveillance and sample collection methods to document freedom from infectious bovine rhinotracheitis in cattle populations

OBJECTIVES: To assess the sensitivity of the current surveillance program used in Denmark for detecting outbreaks of infectious bovine rhinotracheitis (IBR) at the herd level and to evaluate the impact of alternative sample collection strategies on the sensitivity of the system in an acceptable time frame. SAMPLE POPULATION: Data from the Danish Central Husbandry Register on cattle of 24,355 and 25,233 beef herds and on 13,034 and 12,003 dairy herds in the years 2000 and 2001, respectively. PROCEDURES: Surveillance programs were evaluated under current sample collection conditions and under 3 alternative scenarios by use of simulation modeling. Data from the current detection component of the surveillance system were used as input, taking into consideration the sensitivity and specificity of bulktank milk and serologic testing. RESULTS:The current system identifies infected dairy herds within a 3-month period with desired accuracy largely because of the test characteristics and number of bulk-tank milk samples. The system is less likely to detect infected beef herds in a timely manner because surveillance in beef herds depends solely on serologic testing at the time of slaughter. The efficiency of surveillance in dairy cattle herds was not decreased substantially when the slaughter-surveillance component was omitted. CONCLUSIONS AND CLINICAL RELEVANCE:Geographically targeted sample collection during the high-risk season (winter) was predicted to increase the probability of rapid detection of IBR infection in cattle. This approach can be used for assessing other surveillance systems to determine the best strategies for detection of infected herds.     

A retrospective investigation of feline gastrointestinal parasites in western Canada

Between 1998 and 2008, feline fecal specimens were submitted to provincial veterinary diagnostic laboratories in Regina and Saskatoon, Saskatchewan, for sucrose centrifugation-flotation (n = 635), parasite identification (n = 17), and/or Giardia (n = 283) or Cryptosporidium (n = 266) commercial direct immunofluorescence assay (IFA). The most commonly detected parasites on flotation were Toxocara cati (4.7%), Isospora (3.8%), and taeniid eggs (Echinococcus or Taenia) (1.3%). Cats less than 2 years of age were twice as likely to have a positive parasite test as cats older than 2 years. Using IFA, Giardia was detected in 9.9% of samples, and Cryptosporidium in 2.3% of samples. Relative to IFA, flotation had sensitivity values of 39% and 50% for detection of Giardia and Cryptosporidium, respectively. Giardia and Isospora were detected in a higher proportion of samples in our study population than reported in the general cat population in western Canada. This study highlights the importance of sensitivity when interpreting diagnostic tests and provides information to guide region-specific recommendations for helminth parasite prevention and treatment.     

A report of an outbreak of Salmonella oranienburg in a hybrid mouse colony

Mice in a hybrid breeding colony developed symptoms compatible with a diagnosis of salmonellosis. Salmonella oranienburg was recovered from feces by conventional bacteriological techniques. Active salmonellosis appeared to be precipitated by the stress of pregnancy among breeder female BALB/c mice. Necropsy examination of the BALB/c breeder females revealed diarrhea, roughened hair coats, enlarged spleens, white foci in the liver, and skin abscesses. S. oranienburg was recovered from the spleen, cecum, oropharynx, uterus, and skin abscesses of affected mice. DBA/2N breeder males were asymptomatic and S. oranienburg was recovered from the cecum only. Breeding productivity indices dropped to unacceptable levels in rooms containing infected animals. Relatively normal production was observed among uninfected animals of the same strains in adjacent rooms, thus demonstrating the detrimental effect S. oranienburg had on this breeder colony.     

Pre-Hispanic workshop of serpentinite artifacts, venezuelan andes, and possible raw material source

Excavations of a workshop and cemetery in the high Venezuelan Andes yielded winged artifacts made of serpentinite at various stages of finish. Serpentinites are unknown in the Venezuelan Andes, and the raw material was obtained from natural sources located in northern South American or possibly the Antilles, or by trade.     

SPONTANEOUS RESOLUTION OF A PSEUDOANEURYSM IN A HORSE FOLLOWING ANGIOGRAPHIC DIAGNOSIS

Angiography in a horse was used to confirm a pseudoaneurysm of the dorsal metatarsal artery originating from trauma. Gradual clinical resolution of the pseudoaneurysm occurred within days following the procedure. A follow-up angiogram confirmed the clinical impression. An etiology of the spontaneous resolution is presented. Probable influences included technical factors of the injection procedure that caused prolonged presence of the contrast medium within the aneurysmal sac and factors that enhance thrombus formation during angiography.