Isolation and identification of organosulfur compounds oxidizing canine erythrocytes from garlic (Allium sativum)

Five compounds oxidizing canine erythrocytes were isolated from an aqueous ethanol garlic extract by silica gel column chromatography and preparative thin-layer chromatography. On the basis of nuclear magnetic resonance, infrared spectroscopy, and mass spectrometry, they were identified as three known compounds: bis-2-propenyl trisulfide (1), bis-2-propenyl tetrasulfide (2), and bis-2-propenyl pentasulfide (3) as well as two novel compounds, bis-2-propenyl thiosulfonate (4) and trans-sulfuric acid allyl ester 3-allylsulfanyl-allyl ester (5). A mixture of compounds 1-3 and compounds 4 and 5 induced methemoglobin formation in canine erythrocyte suspension in vitro resulting in the oxidation of canine erythrocytes. These groups of characteristic organosulfur compounds contained in garlic probably contribute to oxidations in blood. The constituents of garlic have the potential to oxidize erythrocytes and hemoglobin, suggesting that foods containing quantities of garlic should be avoided for feeding dogs.     

Association between antimicrobial treatment and resistance of pathogenic Escherichia coli isolated from diseased swine in Kagoshima Prefecture,Japan

Pathogenic Escherichia coli is an important cause of diarrhea, edema disease, and septicemia in swine. In Japan, the volume of antimicrobial drugs used for animals is highest in swine, but information about the prevalence of antimicrobial-resistant bacteria is confined to apparently healthy animals. In the present study, we determined the O serogroups, virulence factors, and antimicrobial resistance of 360 E. coli isolates from swine that died of disease in Kagoshima Prefecture, Japan, between 1999 and 2017. The isolates of the predominant serogroups O139, OSB9, O149, O8, and O116 possessed virulence factor genes typically found in diarrheagenic E. coli. We further found five strains of third-generation cephalosporin-resistant E. coli that each produced an extended-spectrum β-lactamase encoded by bla_CTX-M-14, bla_CTX-M-15, bla_CTX-M-24, bla_CTX-M-61, or bla_SHV-12. In 218 swine with a clear history of antimicrobial drug use, we further analyzed associations between the use of antimicrobials for the treatment of diseased swine and the isolation of resistant E. coli. We found significant associations between antimicrobial use and selection of resistance to the same class of antimicrobials, such as the use of ceftiofur and resistance to cefotaxime, cefazolin, or ampicillin, the use of aminoglycosides and resistance to streptomycin, and the use of phenicols and resistance to chloramphenicol. A significant association between antimicrobial use and the resistance of E. coli isolates to structurally unrelated antimicrobials, such as the use of ceftiofur and resistance to chloramphenicol, was also observed.     

Muscle fiber types,growth and development in the whole myotome of cultured Pacific bluefin tuna Thunnus orientalis

Following the successful development of Pacific bluefin tuna (PBT) aquaculture, it is of considerable importance to determine the muscle fiber types and their growth patterns for future development. Muscle fiber profiles of dorsal ordinary, lateral ordinary (LO) and dark muscles and their growth patterns in PBT from 3.0 to 54.3 kg body weight were studied. Muscle fibers were histochemically stained for NADH-diaphorase and myosin adenosine triphosphatase activity (mATPase), and immunohistochemically stained with S-58 slow-muscle myosin antibody. All muscle fibers in dorsal and LO muscles showed low NADH-diaphorase activity, and acid-labile (pH 4.0 or 4.3) and alkali-stable mATPase activity. In LO muscle adjacent to dark muscle, three intensities of mATPase activity were observed after acid pre-incubation at pH 4.5 or 5.0, and the activity was related to the muscle fiber diameter. In dark muscle, all small and some large fibers stained intensely for NADH-diaphorase activity, related to their high aerobic metabolism. The high-active fibers with NADH-diaphorase in dark muscle were positive for S-58 antibody. Some large fibers in dark muscle showed intermediate NADH-diaphorase activity and high mATPase activity after alkali pre-incubations. These are fast-twitch oxido-glycolytic fibers in dark muscle and transformed to red muscle fibers with increasing body weight.     

Pentosan polysulfate regulates hepcidin expression in native Mongolian horses

The aim of this study was to investigate the anti-hepcidin effect of pentosan polysulfate (PPS) in Mongolian horses. Twenty-six healthy horses were randomly allocated in to two-groups; one group was treated with a PPS once a week for 4-weeks while another group keeping as placebo. Blood samples at day 0 (D0), before race (BR; day 28) and after race (AR; day 28) were analyzed for serum biochemistry, hepcidin and iron concentrations. Significant reduction of hepcidin was observed at AR in PPS group when compared with BR placebo (P<0.05) and AR placebo (P<0.01). Mean hepcidin concentration difference of D0-BR and BR-AR in PPS was greater than the placebo whereas the iron concentration difference is reduced compared to placebo. Results indicate a novel therapeutic application of PPS as an anti-hepcidin compound to control hepcidin in horses while emphasizing further molecular studies.     

Detection of nifH Sequences in Sugarcane (Saccharum officinarum L.) and Pineapple (Ananas comosus [L.] Merr.)

To detect N₂-fixing bacteria in a plant without using culture methods, nifH gene segments were amplified with degenerate primers from DNA extracted from stems of sugarcane ( Saccharum officinarum L.) and leaves of pineapple ( Ananas comosus [L.] Merr.). Sequences of the nifH clones were homologous to those of bacteria in the genera Bradyrhizobium, Serratia , and Klebsiella. The bacteria with nifH sequences homologous to those of the three genera may be promising candidates for predominant endophytic diazotrophs in sugarcane and pineapple, because these sequences were commonly detected in samples from different host plants, which were collected at different locations. On the other hand, no nifH sequence related to Gluconacetobacter diazotrophicus , which is an endophytic N₂-fixing bacterium and had been previously isolated from sugarcane, was detected in sugarcane. This indicates the absence or the presence of few G. diazotrophicus in the stems of the sugarcane plants used in the current study.     

Grain qualities and their genetic derivation of 7 new rice for Africa (NERICA) varieties

NERICA are interspecific rice varieties from crossing between the high-yielding Asian rice ( Oryza sativa spp. Japonica) with locally adapted African rice ( Oryza glaberrima). In this study, we analyzed grain qualities of 7 NERICA varieties (NERICA 1 to 7) and genetic derivation of quality-related genes. Quality analyses of NERICA grains showed that 7 NERICA varieties were clearly classified into two groups based on the difference of amylose content, and the difference influenced the pasting and physical properties of grains. Genetic analysis of the gene encoding granule-bound starch synthase I (GBSSI), which is known as a key enzyme on amylose synthesis in rice grain, revealed that varieties with higher amylose content ( approximately 29%) have the gene derived from O. glaberrima parent, and group 2 with lower amylose content ( approximately 22%) have the gene from O. sativa parent. These results indicated that the difference in amylose content as well as grain properties among 7 NERICA varieties is mainly determined by the genetic derivation of GBSSI. Further genetic analysis of starch synthesis-related genes suggested that the genetic derivation of SSIIa also influences the chain length of amylopectin in 7 NERICA varieties.     

Reliable enzyme-linked immunosorbent assay for the determination of coconut milk proteins in processed foods

This study was designed to develop a novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of coconut milk proteins in processed foods. The developed sandwich ELISA was able to detect coconut milk proteins from various coconut milk products and did not show any cross-reactivity with 41 of 42 kinds of popularly used food ingredients, thus reflecting great specificity for coconut milk proteins. In addition, the established ELISA is highly sensitive and allowed the detection of 0.31 μg/g of coconut milk protein in complex food matrices. This proposed assay could serve as a useful tool for the detection of the presence of hidden coconut milk proteins in processed foods.     

Expression of podoplanin in various types of feline tumor tissues

Podoplanin is expressed in various human tumors where it promotes tumor progression, epithelial-mesenchymal transition, and distant metastasis. Podoplanin is also expressed in cancer-associated fibroblasts and induces tumor malignancy. The objective of this study was to evaluate podoplanin expression in various types of feline tumor tissues. Immunohistochemical analysis revealed that podoplanin was expressed in cells of 13/15 (87%) squamous cell carcinomas and 5/19 (26%) fibrosarcomas. Moreover, cancer-associated fibroblasts expressed podoplanin in most tumor types, including 18/21 (86%) mammary adenocarcinoma tissues. Our findings demonstrate that various types of feline tumor tissues expressed podoplanin, indicating the importance of the comparative aspects of podoplanin expression, which may be used as a novel research model for podoplanin biology.     

Relationship of disease progression and plasma histamine concentrations in 11 dogs with mast cell tumors

Plasma histamine concentrations (PHCs) were measured serially over 9 months or until death in 11 dogs with mast cell tumors (MCTs). Eight dogs had grossly visible disease and the other 3 dogs had microscopic disease. Initial PHCs in the dogs with gross disease were significantly higher than PHCs in healthy dogs (median, 0.73 ng/mL and 0.19 ng/mL respectively; P < .009), whereas initial PHCs in dogs with microscopic disease showed no difference from controls. Seven dogs subsequently had progressive increases in PHC, and developed hyperhistaminemia (median, 14.0 ng/mL; range, 5.11-30.1 ng/nL). These 7 dogs died from MCTs, and 1 had general weakness with rapid lysis of a large tumor burden after radiation therapy. PHCs of the other 4 dogs were less than 1 ng/mL during the study. These 4 dogs were still alive with adequate control of the tumor at the conclusion of the study. Four of the 11 dogs initially had gastrointestinal (G1) signs, which abated soon after administration of histamine-2 (H-2) blockers. No significant difference was found between PHCs in dogs with GI signs and those without GI signs (median, 0.86 ng/mL and 0.35 ng/mL. respectively). Thereafter, 7 dogs had serious GI complications for which H-2 blocker therapy was ineffective. PHCs in these 7 dogs were extremely high (median, 12.2 ng/mL; range, 3.42-30.1 ng/nL). Results of this study demonsrated that PHC was one factor related to disease progression, and indicated that marked hyperhistaminemia was associated with the GI signs refractory to H-2 blocker therapy in dogs with MCTs.     

An SNP caused loss of seed shattering during rice domestication

Loss of seed shattering was a key event in the domestication of major cereals. We revealed that the qSH1 gene, a major quantitative trait locus of seed shattering in rice, encodes a BEL1-type homeobox gene and demonstrated that a single-nucleotide polymorphism (SNP) in the 5' regulatory region of the qSH1 gene caused loss of seed shattering owing to the absence of abscission layer formation. Haplotype analysis and association analysis in various rice collections revealed that the SNP was highly associated with shattering among japonica subspecies of rice, implying that it was a target of artificial selection during rice domestication.