Real-time PCR genotyping assay for feline erythrocyte pyruvate kinase deficiency and mutant allele frequency in purebred cats in Japan

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.     

Hypoxia inducible factor 1α expression and effects of its inhibitors in canine lymphoma

Hypoxic conditions in various cancers are believed to relate with their malignancy, and hypoxia inducible factor-1α (HIF-1α) has been shown to be a major regulator of the response to low oxygen. In this study, we examined HIF-1α expression in canine lymphoma using cell lines and clinical samples and found that these cells expressed HIF-1α. Moreover, the HIF-1α inhibitors, echinomycin, YC-1 and 2-methoxyestradiol, suppressed the proliferation of canine lymphoma cell lines. In a xenograft model using NOD/scid mice, echinomycin treatment resulted in a dose-dependent regression of the tumor. Our results suggest that HIF-1α contributes to the proliferation and/or survival of canine lymphoma cells. Therefore, HIF-1α inhibitors may be potential agents to treat canine lymphoma.     

Lipopolysaccharide induces expression of collagen VI in the rat lung

The involvement of the lung during the septic systemic inflammatory response elicited by administration of lipopolysaccharide (LPS) was investigated. Eight-week-old male Sprague–Dawley rats were injected i.p. with 15 mg/kg LPS. After 24 h, the lungs were excised to evaluate the cellular responses to LPS. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) analysis revealed that type VI collagen (ColVI) was extremely upregulated during sepsis in the rat lung within the first 24 h of LPS administration. Upregulation of ColVI protein and its mRNA was demonstrated by Western blot analysis, real time PCR, and immunohistochemistry. To the best of our knowledge, this is the first report demonstrating the activation of ColVI in the rat lung at the early stage of systemic inflammation. Activation of ColVI might be involved in sepsis-mediated lung fibrosis at an early stage.     

Importance of shingle beaches as habitat for Manila clam Ruditapes philippinarum

Fisheries Science - We carried out surveys to clarify how the density (no. of individuals/m2) and condition factor (CF) of Manila clam Ruditapes philippinarum differ between estuarine tidal flats...     

Pilot Study to Examine Radial Variation in Annual Ring Width, Density and Shrinkage

The annual ring width, density and shrinkage variation from pith to bark in Chinese fir (Cunninghamia lanceolata) and Boka sugi (Cryptomeria japonicd) were studied and compared. The results show that the ring width decreased sharply from pith to bark for Chinese fir. However, the ring width variation pattern for Boka sugi followed a different way, i.e., the ring width decreased to the fifth ring, increased to the tenth ring, decreased again to the fifteenth, and then increased to the twentieth, where it became constant. The large variations of Boka sugi appeared to show the maintenance of fast growth for many years. The annual ring mean density of Chinese fir increased gradually from pith to bark. However, the density changes for Boka sugi indicated the opposite trend, i.e., the mean density decreased gradually from pith to bark. The former showed a pattern as the same as a pine and a larch, and the latter was often found in a cedar and a cypress. The longitudinal shrinkage in juvenile wood was much hig     

The presence of double-stranded RNAs in <Emphasis Type="Italic">Alternaria alternata</Emphasis> Japanese pear pathotype is associated with morphological changes

Strains of the Japanese pear pathotype of Alternaria alternata were screened for double-stranded RNAs (dsRNAs). Four strains had several dsRNAs; strain N18 was associated with several dsRNAs and had impaired growth phenotypes such as irregular mycelium and abnormal pigmentation. We isolated dsRNA-cured isolates from strain N18 by single-conidium isolation. The dsRNA-cured isolates had recovered normal growth and pigmentation. Enlarged vesicles were observed in mycelial cells of the original dsRNA-carrying N18 strain. DAPI nuclear staining revealed regression of the nuclei in dsRNA-carrying N18 cells. These results indicate that the dsRNAs might have negative effects, such as apoptosis-like cell death, on the host fungus.     

Bow variation in kiln-dried boxed-heart square timber of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

To evaluate the bow variation in boxed-heart square timber of sugi (Cryptomeria japonica), bows from kiln-dried timber for five sugi cultivars with different longitudinal shrinkage trends were compared for two stem heights. Two general trends were observed, depending on the cultivar: (1) either the bow was larger at the lower than at the upper part of the stem, or (2) the bows at the lower and the upper parts of the stem were similar. In timber that had larger bow values, the gradients of longitudinal shrinkage were large across the radius and along the length of the timber. There was a positive relationship between the bow and longitudinal shrinkage. These results suggest that the bow variation between the timbers was caused by a variation in longitudinal shrinkage, which was affected by the microfibril angle. Furthermore, the bow was inversely proportional to the modulus of elasticity, which suggests that timber with a low modulus of elasticity is susceptible to a large bow due to large longitudinal shrinkage.     

Tangential and radial shrinkage variation within trees in sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

The transverse shrinkage variation within trees was examined for five sugi cultivars. The within-tree trends of tangential shrinkage (α _T) were different by cultivar, whereas radial shrinkage (α _R) increased from pith to bark in most cultivars. The tangential/radial shrinkage ratio (α _T/α _R) decreased from pith to bark in most cultivars, because the radial variation of α _R was larger than that of α _T. The cultivars showed significant differences among cultivars in α _T, α _R, and α _T/α _R, but the difference among cultivars for α _T/α _R was smaller. The relationships between transverse shrinkage and microfi bril angle (MFA), basic density (BD), tree ring parameters, and modulus of elasticity were examined. The α _T and α _R showed positive relationships with BD, latewood percentage, latewood density, and modulus of elasticity, and negative relationships with MFA and ring width. The relationships with earlywood density were weak. Sugi exhibited variation in transverse shrinkage within stem and among cultivars, with the variation affected by MFA, density, and tree ring parameters. Part of this article was presented at the 56th Annual Meeting of the Japan Wood Research Society, Akita, Japan, August 2006, and the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, August 2007     

Biological and chemical stability of garlic-derived allicin

This study verifies the instability of garlic ( Allium sativum L.)-derived allyl 2-propenylthiosulfinate (allicin) in various aqueous and ethanolic solutions as well as in vegetable oil through chemical and biological analyses performed simultaneously. Crushed fresh garlic cloves generated antibacterial activity and chemically detectable allicin, a major antibacterial principle, and both declined on a daily basis in aqueous and ethanolic solutions at room temperature, showing biological and chemical half-lives of about 6 and 11 days, respectively. Allicin was more stable in 20% alcohol than in water, but surprisingly unstable in vegetable oil, with an activity half-life 0.8 h, as estimated from its antibacterial activity toward Escherichia coli, and a chemical half-life of 3.1 h, based on chromatographic quantification. In alcoholic and aqueous extracts, the biological half-life of allicin tended to be longer than the chemical one, suggesting the occurrence of bioactive compounds other than allicin in the extracts.     

Prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma

Currently, antimicrobial-resistant staphylococci, particularly methicillin-resistant Staphylococcus pseudintermedius (MRSP), are frequently isolated from canine superficial pyoderma in Japan. However, little is known regarding the nasal prevalence of MRSP in pet dogs. Here, we determined the prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma. Of the 125 nares and 108 affected sites of pet dogs with superficial pyoderma, 107 (13 species) and 110 (eight species) staphylococci strains, respectively, were isolated. The isolation rate of S. pseudintermedius from pyoderma sites (82/110 strains, 74.5%) was significantly higher than that from nares (57/107 strains, 53.3%) (P<0.01). Notably, the prevalence of MRSP (18/57 strains, 31.6%) in nares was equivalent to that in pyoderma sites (28/82 strains, 34.1%). Furthermore, the phenotypes and genotypes of antimicrobial resistance in MRSP strains from nares were similar to those from pyoderma sites. Our findings revealed that the prevalence of antimicrobial-resistant staphylococci in the nares of pet dogs with superficial pyoderma is the same level as that in affected sites. Therefore, considerable attention should be paid to the antimicrobial resistance of commensal staphylococci in companion animals.