A critical role for the innate immune signaling molecule IRAK-4 in T cell activation

IRAK-4 is a protein kinase that is pivotal in mediating signals for innate immune responses. Here, we report that IRAK-4 signaling is also essential for eliciting adaptive immune responses. Thus, in the absence of IRAK-4, in vivo T cell responses were significantly impaired. Upon T cell receptor stimulation, IRAK-4 is recruited to T cell lipid rafts, where it induces downstream signals, including protein kinase C activation through the association with Zap70. This signaling pathway was found to be required for optimal activation of nuclear factor kappaB. Our findings suggest that T cells use this critical regulator of innate immunity for the development of acquired immunity, suggesting that IRAK-4 may be involved in direct signal cross talk between the two systems.     

Characterization of wood-based molding bonded with citric acid

The wood-based moldings were fabricated by using only citric acid as an adhesive. The mechanical properties, water resistances, thermal properties and chemical structure were investigated. Wood powder obtained from Acacia mangium was mixed with citric acid under certain weight ratios (0–40 wt%), and each powder mixture was molded using two types of metal molds at 200°C and 4 MPa for 10 min. The modulus of rupture and the modulus of elasticity values of the wood-based molding containing 20 wt% citric acid were 35.8 MPa and 5.4 GPa, respectively. The maximum impact strength was 0.94 kJ/m² with the same citric acid content. The water resistance increased with increasing citric acid content, and the good resistance to boiling water was also recognized. However, some elution of substances derived from citric acid was observed at high levels of citric acid, causing a decrease in the thermal properties. The ester linkages were detected by Fourier transform infrared spectroscopy, indicating that the citric acid reacted with the wood. Our results demonstrated that citric acid brought about good adhesiveness for fabricating the wood-based molding.     

Comparison of oral robenacoxib and ketoprofen for the treatment of acute pain and inflammation associated with musculoskeletal disorders in cats: A randomised clinical trial

The objective of the study was to evaluate the efficacy and tolerability of robenacoxib, a selective cyclooxygenase-2 inhibitor, for the treatment of acute pain and inflammation associated with musculoskeletal disorders in cats. The study was a prospective, multi-centre, randomised, blinded, non-inferiority design clinical trial comparing robenacoxib to ketoprofen. A total of 68 cats presenting with pain and inflammation associated with acute musculoskeletal disorders were recruited and allocated randomly to receive, orally once daily for 5-6days, either 1.0-2.4mg/kg robenacoxib (n=47) or 1mg/kg ketoprofen (n=21). The primary efficacy endpoint was the total clinician score, which was the sum of clinician numerical rating scale scores for pain, inflammation and mobility. Assessments were made at baseline, on day 2, and day 4 or 5. For the total clinician score, non-inferior efficacy of robenacoxib was demonstrated with a relative efficacy of 1.151 (95% confidence interval 0.872-1.494). Non-inferior efficacy of robenacoxib was also demonstrated for the secondary endpoint of the total owner score. Robenacoxib was superior (P<0.05) to ketoprofen for the owner's assessment of activity and human/animal relationship. The tolerability of both treatments was good as assessed by monitoring adverse events, clinical signs and haematology and serum biochemistry variables.     

Analysis of preference for domestic grass‐fed beef in Japanese youths

A questionnaire based on sensory evaluation of completely domestic grass‐fed beef was carried out on 157 Japanese undergraduate students aged between 18 and 22 years in Kitasato University. The sensory evaluation sheet consisted of 10 questions concerning preference for domestic grass‐fed beef, and three demographic/lifestyle questions. Using principal component analysis and cluster analysis, the respondents were divided into four groups (G1–G4). G1 accepted almost all properties. G2 accepted most properties but disliked chewiness. G3 accepted juiciness and flavor but disliked the color and texture of the meat. G4 tended to dislike almost all properties. According to chi‐square test, most G2‐people statistically liked other commercial beef and G4‐people had neutral and negative impressions. G1‐ or G3‐people did not have any significant tendency as regards beef preference. These results indicate that most of the young respondents who preferred domestic grass‐fed beef could not accept its texture, and some respondents could accept its juiciness and flavor. It is also suggested that a part of the people who like commercial beef do not prefer chewiness of grass‐fed beef. Such information will aid grass‐fed beef cattle breeders, producers and packers to improve the quality of beef and its evaluation.     

Isolation and molecular characterization of multidrug-resistant Gram-negative bacteria from imported flamingos in Japan

Imported animals, especially those from developing countries, may constitute a potential hazard to native animals and to public health. In this study, a new flock of lesser flamingos imported from Tanzania to Hiroshima Zoological Park were screened for multidrug-resistant Gram-negative bacteria, integrons and antimicrobial resistance genes. Thirty-seven Gram-negative bacterial isolates were obtained from the flamingos. Seven isolates (18.9%) showed multidrug resistance phenotypes, the most common being against: ampicillin, streptomycin, tetracycline, trimethoprim/sulfamethoxazole and nalidixic acid. Molecular analyses identified class 1 and class 2 integrons, β-lactamase-encoding genes, bla_TEM-1 and bla_CTX-M-2 and the plasmid-mediated quinolone resistance genes, qnrS and qnrB. This study highlights the role of animal importation in the dissemination of multidrug-resistant bacteria, integrons and antimicrobial resistance genes from one country to another.     

Purification and characterization of cathepsin S from hepatopancreas of carp Cyprinus carpio

SUMMARY: Cathepsin S was purified from carp hepatopancreas to homogeneity up to 300-fold. The amino acid sequence of its NH₂-terminus was determined to be V-P-D-A-M-D-W-Y-N-K-G-Y-V-T-D-V-K-N-Q. On the contrary, that of purified cathepsin L from carp hepatopancreas was to be V-P-N-S-L-D-W-R-E-K-G. Purified cathepsin S consisted of a single chain with 37 kDa estimated by sodium dodecylsulfate–polyacrylamide gel electrophoresis. The enzyme had strong hydrolytic activity toward Z-Phe-Arg-MCA with the pH optimum of 7.0, but this lacked the ability to hydrolyze most of the other MCA substrates. The optimum pH of cathepsin S for protein substrate (carp myosin heavy chain) was also to be pH 7.0. These properties of purified cathepsin S obviously differ from cathepsins B and L. The enzyme activity was totally inhibited by E-64, leupeptin, 5–5'-dithiobis (2-nitro-benzoic acid) and p -tosyl-lys chloromethylketone as well.     

Genotyping of Giardia intestinalis from domestic and wild animals in Japan using glutamete dehydrogenase gene sequencing

To determine the genotypes of Giardia intestinalis from domestic and wild animals in Japan, Giardia isolates obtained from feces of 24 dogs kept in households and breeding kennels, three companion cats, five dairy calves and three wild monkeys, Macaca fuscata, were genotyped using the 177 bp sequence of the glutamete dehydrogenase gene (gdh). The genotypes were assemblages A, C, D or A/D for dog isolates, Assemblage F for cat isolates, assemblages A or E for calf isolates and assemblage B for monkey isolates. This is the first report on the genotypes of Giardia isolates from cats, calves and wild monkeys in Japan.     

Induction of immune responses against glycosphingolipid antigens: comparison of antibody responses in mice immunized with antigen associated with liposomes prepared from various phospholipids

The immune responses of mice against glycosphingolipid (GSL) antigens and the effect of the phospholipid composition of liposomes on the immunogenicity in mice of liposome-associated GSL antigens were examined. The immunization with GSL antigen alone was unable to induce any detectable anti-GSL antibody responses. On the other hand, the immune responses against GSL antigens were detected after immunization with liposomes composed of dipalmitoylphosphatidylcholine (DPPC) (0.5 micromol), cholesterol (Chol) (0.5 micromol), Salmonella minnesota R595 lipopolysaccharides (LPS) (10 microg) and GSL (0.05 micromol) (DPPC-liposome). However, the administration with liposome composed of dimyristoylphosphatidylcholine (DMPC) (0.5 micromol), Chol (0.5 micromol), S. minnesota R595 LPS (10 microg) and GSL (0.05 micromol) and with liposomes composed of distearylphosphatidylcholine (DSPC) (0.5 micromol), Chol (0.5 micromol), and S. minnesota R595 LPS (10 microg) and GSL (0.05 micromol) was ineffective for the induction of the immune responses against GSL antigens. These results suggest that DPPC-liposome would serve effectively as a delivery vehicle for inducing immune responses against GSL antigen.     

Molecular characterization of parthenogenic Fasciola sp. in Korea on the basis of DNA sequences of ribosomal ITS1 and mitochondrial NDI gene

Nucleotide sequences of ribosomal internal transcribed spacer (ITS1) and mitochondrial NADH dehydrogenase I (NDI) gene were analyzed to genetically characterize aspermic Fasciola forms in Korea. From the difference in ITS1 sequences, Korean flukes were divided into 3 haplotypes represented by Kor1, Kor2 and Kor1/2, which had nucleotides identical to F. hepatica, F. gigantica and those overlapped between the two species, respectively. NDI sequences also showed that Korean flukes could be classified into 3 distinct haplotypes (Kor1: F. hepatica-type, Kor2a and Kor2b: F. gigantica-type). The sequences of Kor1 and Kor2a were 100% identical to those of the haplotypes Fsp1and Fsp2, respectively, which are major Fasciola forms in Japan. These findings strongly suggest that aspermic Fasciola forms in Korea and Japan originated from same ancestors and have recently spread throughout both countries.     

Quantitative in vivo measurement of central benzodiazepine receptors in the brain of cats by use of positron-emission tomography and [11C]flumazenil

OBJECTIVE: To map central benzodiazepine receptors (BZRs) in the brain of cats by use of positron-emission tomography (PET) and [11C]flumazenil. ANIMALS: 6 male cats that weighed between 2.0 and 3.6 kg. PROCEDURE: Brain images obtained by PET evaluation of [11C]flumazenil were superimposed on T2-weighted magnetic-resonance imaging (MRI) scans of the same cats. Detailed anatomic regions, such as the cerebral cortex, striatum, thalamus, midbrain, and cerebellum, on the PET images were evident by PET-MRI registration. Regional binding of [11C]flumazenil to BZRs was quantitatively measured by use of a model with 2 tissue compartments and 4 variables. RESULTS: The highest value for distribution volume was observed in the cerebral cortex, and the lowest value was found in the midbrain of cats. CONCLUSIONS AND CLINICAL RELEVANCE: Binding of [11C]flumazenil to BZRs in the brain of cats can be quantitatively measured by use of PET with the aid of PET-MRI registration. It is difficult to diagnose changes in these neuroreceptors within the field of current veterinary science. In the future, PET should prove useful for investigating and diagnosing brain disorders in animals in clinical settings.