The dynamics of antimicrobial-resistant Campylobacter jejuni on Japanese broiler farms

We investigated for dynamics of Campylobacter clones on 2 different managerial broiler farms. Campylobacter isolates were differentiated by resistance typing and molecular typing methods. On farm I, the same C. jejuni clones resistant to fluoroquinolone and oxytetracycline were isolated after one and half years again and another susceptible clone was invaded. The susceptible clone was isolated again after half year. Broiler flocks on the farm may be repeatedly infected with a few C. jejuni clones. On farm II, new clones including antimicrobial resistant one, were often invaded. The change of predominant C. jejuni clone in each flock on both the farms was observed, in the absence of antimicrobial selective pressure.     

Chlorogenic acid in coffee can prevent the formation of dinitrogen trioxide by scavenging nitrogen dioxide generated in the human oral cavity

Coffee contains antioxidants like chlorogenic acid and its isomers. In this report, effects of coffee on the nitrite-induced N2O3 formation were studied using whole saliva and bacterial fraction prepared from the saliva. The formation of N2O3 was measured by fluorescence increase due to the transformation of 4,5-diaminofluorescein to triazolfluorescein. Coffee inhibited the nitrite-induced fluorescence increase, and 50% inhibition was observed at several microg of coffee/mL in bacterial fraction of saliva as well as whole saliva. During the inhibition of the fluorescence increase, concentration of chlorogenic acid and its isomers decreased. It is discussed that the reduction of NO2 by chlorogenic acid and its isomers contributed to the coffee-dependent inhibition of the fluorescence increase as N2O3 is formed from NO and NO2. When coffee was added to whole saliva, chlorogenic acid and its isomers bound to cells in the saliva. The rate of the fluorescence increase in bacterial fraction, which was prepared at defined periods after the ingestion of coffee, was increased to the rate before the ingestion of coffee with a half-time of about 1 h. This result suggests that chlorogenic acid and its isomers remained in the oral cavity for a few hours after ingestion of coffee. The significance of coffee drinking and rinsing of the mouth with coffee for the health of the oral cavity is proposed.     

DaizuBase,an integrated soybean genome database including BAC-based physical maps

Soybean [Glycine max (L) Merrill] is one of the most important leguminous crops and ranks fourth after to rice, wheat and maize in terms of world crop production. Soybean contains abundant protein and oil, which makes it a major source of nutritious food, livestock feed and industrial products. In Japan, soybean is also an important source of traditional staples such as tofu, natto, miso and soy sauce. The soybean genome was determined in 2010. With its enormous size, physical mapping and genome sequencing are the most effective approaches towards understanding the structure and function of the soybean genome. We constructed bacterial artificial chromosome (BAC) libraries from the Japanese soybean cultivar, Enrei. The end-sequences of approximately 100,000 BAC clones were analyzed and used for construction of a BAC-based physical map of the genome. BLAST analysis between Enrei BAC-end sequences and the Williams82 genome was carried out to increase the saturation of the map. This physical map will be used to characterize the genome structure of Japanese soybean cultivars, to develop methods for the isolation of agronomically important genes and to facilitate comparative soybean genome research. The current status of physical mapping of the soybean genome and construction of database are presented.     

Substrate specificity of carp hepatopancreatic cathepsin S for peptides and myofibrillar proteins

SUMMARY: From 250 g of carp hepatopancreas, 0.29 mg of the purified enzyme was obtained. The bond specificity of cathepsin S for α-neoendorphin and neurotensin was ⁶Arg-⁷Lys and ³Tyr-⁴Glu, respectively. The cleavage sites for insulin B-chain were estimated to be the bonds at ³Asn-⁴Gln, ⁶Leu-⁷Cys, ¹²Val-¹³Glu, ¹³Glu-¹⁴Ala, ¹⁶Tyr-¹⁷Leu, ²²Arg-²³Gly, ²⁴Phe-²⁵Phe and ²⁶Tyr-²⁷Thr. P2 position on these peptides were bulky and hydrophobic amino acid residues such as Phe or Leu, small amino acid residues such as Gly, Ala and Val were also accepted in these positions. Regarding the protein substrates, cathepsin S degraded carp α-actinin, actin, tropomyosin and troponins T and I although the proteolyzing speeds were distinct from one another.     

Genome-wide association mapping focusing on a rice population derived from rice breeding programs in a region

Plant breeding programs in local regions may generate genetic variations that are desirable to local populations and shape adaptability during the establishment of local populations. To elucidate genetic bases for this process, we proposed a new approach for identifying the genetic bases for the traits improved during rice breeding programs; association mapping focusing on a local population. In the present study, we performed association mapping focusing on a local rice population, consisting of 63 varieties, in Hokkaido, the northernmost region of Japan and one of the northern limits of rice cultivation worldwide. Six and seventeen QTLs were identified for heading date and low temperature germinability, respectively. Of these, 13 were novel QTLs in this population and 10 corresponded to the QTLs previously reported based on QTL mapping. The identification of QTLs for traits in local populations including elite varieties may lead to a better understanding of the genetic bases of elite traits. This is of direct relevance for plant breeding programs in local regions.     

Improved separation method for highly purified lutein from Chlorella powder using jet mill and flash column chromatography on silica gel

We investigated an improved method for the separation of high-purified lutein from a commercially available spray-dried Chlorella powder (CP) using fine grinding by jet mill and flash column chromatography on a silica gel. Saponification and extraction of lutein were enhanced 2.3-2.9-fold in jet mill-treated CP (mean particle size, 20 microm) as compared to untreated CP (mean particle size, 67 microm). The carotenoid extract was dissolved in ether-hexane (1:1 v/v) and subjected to flash column chromatography on silica gel. A mixture of alpha- and beta-carotene was eluted with hexane, followed by elution with hexane-acetone-chloroform (7:2:1 v/v). Lutein (dark-orange band) was collected after the elution of an unknown colorless compound (detected based on UV absorbance). The purity of lutein in this fraction was over 99%, and the yield was 60%. The present study provides key information for obtaining highly purified lutein using flash column chromatography on a silica gel.     

The pharmacokinetics of primaquine in calves after subcutaneous and intravenous administration

The pharmacokinetics of primaquine was studied in calves of 180–300 kg live weight. Primaquine was injected at 0.29 mg/kg (0.51 mg/kg as primaquine diphosphate) intravenously (IV) or subcutaneously (SC) and the plasma concentrations of primaquine and its metabolite carboxyprimaquine were determined by high-performance liquid chromatography. The extrapolated concentration of primaquine at zero time after IV administration was 0.50±0.48 µg/ml (mean ±SD) which decreased with an elimination half-life of 0.16±0.07 h. Primaquine was rapidly converted to carboxyprimaquine after either route of administration. The peak concentration of carboxyprimaquine was 0.50±0.08 µg/ml at 1.67±0.15 h after IV administration. The corresponding value was 0.47±0.07 µg/ml at 5.05±1.20 h after SC administration. The elimination half-lives of carboxyprimaquine after IV and SC administration were 15.06±0.99 and 12.26±3.06 h, respectively. The areas under the concentration-time curve for carboxyprimaquine were similar following either IV or SC administration of primaquine; the values were 11.85±2.62 µg.h/ml after the former and 10.95±2.65 µg.h/ml after the latter. The mean area under the concentration-time curve for primaquine was less than 0.1 µg.h/ml after either route of administration.Abbreviations AUC area under the concentration-time curve - CPRQ carboxyprimaquine - IV intravenous - 6M8AQ 6-methoxy-8-aminoquinoline - PRQ primaquine - SC subcutaneous     

Assessment of the factorial structures of the C-BARQ in Japan

In order to introduce the Canine Behavioral Assessment and Research Questionnaire (C-BARQ), which is a standardized system for evaluating the behavioral traits of dogs, to Japan, where the environment with respect to dog ownership is thought to differ from those of the United States and Europe, we compared demographic information on dogs in the United States and Japan and examined whether similar factors could be extracted from both countries using questionnaire items of the C-BARQ. The C-BARQ was completed by 11,410, and 734 dog owners respectively in the United States and Japan, and some demographic differences were found, such as breed and neuter status. Data from completed questionnaires were subjected to factor analysis, and the resulting factors were tested for reliability. In the United States, factor analysis yielded 11 factors from 63 items that accounted for 52.9% of the common variance. In Japan, 15 factors were extracted, and these accounted for 57.0% of the common variance. The present factors for the United States were almost identical to the factors identified in a previous study, and similar factors were extracted in both countries. Therefore, the C-BARQ can function effectively as a fundamental behavioral evaluation system for dogs in Japan.