A major genome region underlying artemisinin resistance in malaria

Evolving resistance to artemisinin-based compounds threatens to derail attempts to control malaria. Resistance has been confirmed in western Cambodia and has recently emerged in western Thailand, but is absent from neighboring Laos. Artemisinin resistance results in reduced parasite clearance rates (CRs) after treatment. We used a two-phase strategy to identify genome region(s) underlying this ongoing selective event. Geographical differentiation and haplotype structure at 6969 polymorphic single-nucleotide polymorphisms (SNPs) in 91 parasites from Cambodia, Thailand, and Laos identified 33 genome regions under strong selection. We screened SNPs and microsatellites within these regions in 715 parasites from Thailand, identifying a selective sweep on chromosome 13 that shows strong association (P = 10(-6) to 10(-12)) with slow CRs, illustrating the efficacy of targeted association for identifying the genetic basis of adaptive traits.     

Age‐Dependent Morphology of NADPH Diaphorase‐Positive Amacrine Cells in the Mouse Retina

NADPH diaphorase‐positive amacrine cells (NAC) were studied in retinal whole mount preparation of mice, ranging from 1 day to 30 months of age. Following a peak in number and size during early development at postnatal day 14, their number and distribution remained well preserved up to senescence. Functional considerations include immunological, vascular and neuro‐modulating aspects.     

Effects of sialidase knockout and complementation on virulence of Mycoplasma gallisepticum

Reannotation of the pathogenic Mycoplasma gallisepticum strain R(low) genome identified the hypothetical gene MGA_0329 as a homolog of the sialidase gene MS53_0199 of Mycoplasma synoviae strain MS53. Potent sialidase activity was subsequently quantitated in several M. gallisepticum strains. Because sialidase activity levels correlate significantly with differing M. synoviae strain virulence, we hypothesized this enzyme may also influence the virulence of M. gallisepticum. MGA_0329 was disrupted in strain R(low) to create mutants 6, 358 and P1C5, which resulted in the loss of sialidase activity in all three mutants. Chickens infected with the knockout mutants had significantly less severe (P<0.05) tracheal lesions and tracheal mucosal thickening than chickens infected with equal doses of strain R(low). Significantly fewer (P<0.05) CCU especially of strains 6 and P1C5 were recovered at necropsy. Mini-Tn4001tet plasmid pTF20 carrying a wild-type copy of MGA_0329 with its native promoter was used to complement the genetic lesion in strain P1C5. Three clones derived from P1C5, each having one copy of MGA_0329 stably transposed into a different site in its genome, expressed sialidase restored to wild-type activity levels (1.58×10(-8)U/CFU). Complementation of P1C5 with MGA_0329 did not restore it to wild-type levels of virulence, indicating that the contribution of sialidase to M. gallisepticum virulence is not straightforward.     

Effects of a mock ultrasonographic procedure on cortisol concentrations during low-dose dexamethasone suppression testing in clinically normal adult dogs

OBJECTIVE: To determine whether the stress of an ultrasonographic procedure would interfere with the suppressive effect of dexamethasone during a low-dose dexamethasone suppression test (LDDST) in healthy dogs. ANIMALS: 6 clinically normal adult dogs. PROCEDURE: In phase 1, an LDDST was performed 5 times at weekly intervals in each dog. Serum samples were obtained 0, 2, 4, 6, and 8 hours after dexamethasone injection. A mock 20-minute abdominal ultrasonographic examination was performed on all dogs at each time point during the LDDST on weeks 2 through 5. In phase 2, serum cortisol concentrations were measured before and immediately after a 20-minute mock abdominal ultrasonographic examination, as described for phase 1. RESULTS: We did not detect significant differences after dexamethasone injection when comparing median cortisol concentrations for weeks 2 to 5 (mock ultrasonographic procedure) with median concentration for week 1 (no mock ultrasonographic procedure). For 5 of the 6 dogs, cortisol concentrations after dexamethasone injection decreased to < 35.9 nmol/L after each mock ultrasonographic procedure and remained low for the duration of the LDDST. In phase 2, all dogs had significant increases in cortisol concentrations immediately after the mock ultrasonographic procedure. CONCLUSIONS AND CLINICAL RELEVANCE: A 20-minute mock abdominal ultrasonographic examination performed during LDDST did not alter results of the LDDST in most dogs. Cortisol concentrations measured immediately after a mock ultrasonographic examination were significantly increased. Ultrasonographic procedures should be performed a minimum of 2 hours before collection of samples that will be used to measure cortisol concentrations.     

Biochemical characterisation of navicular hyaline cartilage,navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease

The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition.     

Experimental evaluation of urinary bladder marsupialization in male goats

OBJECTIVE: To evaluate the outcome of urinary bladder marsupialization in male goats. STUDY DESIGN: Prospective, experimental study. ANIMALS: Six healthy mixed-breed male goats. METHODS: After experimentally induced urethral obstruction, 6 male goats had urinary bladder marsupialization. Renal ultrasonography, complete blood count, and serum biochemical analysis were evaluated preoperatively (day 0), at 7 postoperative days, and then at 30-day intervals until 180 days. Stomal diameter was recorded immediately postoperatively and at each postoperative interval. Necropsy examination was performed on day 180 or when stomal stricture or death occurred. RESULTS: Stomal stricture occurred in 1 goat at 120 days. Another goat was found dead at 150 days; severe, suppurative cystitis was identified on necropsy. All goats had mild urine scald dermatitis. Serum biochemical values remained within normal limits, but significant decreases in white blood cell count, serum creatinine concentration, and stomal diameter occurred. At necropsy, all bladders were tubular in shape. Histological evidence of chronic suppurative cystitis and chronic, mild lymphoplasmacytic pyelitis occurred in all goats. Bacterial culture of renal tissue yielded growth in 3 goats, and bladder mucosal swabs yielded bacterial growth in all goats. CONCLUSIONS: Although clinical signs of ascending urinary tract infection were not observed in goats with patent stomata, urinary bladder marsupialization may result in ascending urinary tract inflammation or infection. CLINICAL RELEVANCE: Based on our results, urinary bladder marsupialization should be recommended with caution as the primary method for management of urinary tract obstruction in clinical cases.     

Prevalence and importance of internal tandem duplications in exons 11 and 12 of c-kit in mast cell tumors of dogs

OBJECTIVE: To determine the prevalence of activating internal tandem duplications (ITDs) in exons 11 and 12 of c-kit in mast cell tumors (MCTs) of dogs and to correlate these mutations with prognosis. SAMPLE POPULATION: 157 formalin-fixed, paraffin-embedded MCTs from dogs in the pathology database of the Veterinary Medical Teaching Hospital at the University of California, Davis. PROCEDURE: Genomic DNA was isolated from tumor specimens and a polymerase chain reaction procedure was performed to determine whether there were ITDs in exons 11 and 12. RESULTS: We identified ITDs in 1 of 12 (8%) grade-I, 42 of 119 (35%) grade-lI, and 9 of 26 (35%) grade-ll tumors (overall prevalence, 52 of 157 [33%]). Logistic regression analysis revealed that the odds of grade-II and -III tumors possessing an ITD were approximately 5 times greater than that for grade-I tumors, although these odds did not differ significantly. Although MCTs possessing an ITD were twice as likely to recur after excision and twice as likely to result in metastasis as those without an ITD, these values also did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE: These results provide evidence that ITDs in c-kit occur frequently in MCTs of dogs. The high prevalence of c-kit activating mutations in MCTs of dogs combined with the relative abundance of mast cell disease in dogs provide an ideal naturally developing tumor in which to test the safety and efficacy of novel small-molecule kinase inhibitors such as imatinib mesylate.     

Molecular diversity of barley yellow dwarf virus-PAV from China and the Czech Republic

Wheat yellow dwarf disease (BYD), caused by different species of barley/cereal yellow dwarf viruses (B/CYDVs), is one of the most serious cereal diseases in China and the Czech Republic. Because genetic diversity of the virus directly influences disease epidemiology, the molecular diversity and population structure of 24 Chinese isolates and 16 the Czech Republic isolates of BYDV-PAV from different regions in two countries were analyzed by sequencing their coat protein (CP) and readthrough protein (RTP) domain (RTD) genes and comparing the sequences with six CP and 16 RTP sequences of BYDV-PAV isolates from the NCBI database based on nucleotide identity position, phylogenetic analysis and nucleotide diversity. Nucleotide identities between the Chinese and the Czech Republic isolates for the CP were 76.6–99.4%, 73.9–89.1% for RTD (ORF5), respectively. The Chinese and the other country isolates showed 74.7–99.2% nucleotide identity for RTP (ORF3+ORF5). Phylogenetic analysis of CP sequences showed that 20 Chinese isolates clustered in the same clade, but the other four Chinese isolates clustered in another clade with the isolates from the Czech Republic and other counties. The population of BYDV-PAV in China had greater nucleotide variability and was more divergent than that in the Czech Republic. Geographical and ecological factors but not hosts might contribute to the population differences in the two countries.