Isolation of a Megatrypanum trypanosome from sika deer (Cervus nippon yesoensis) in Japan

A trypanosome was isolated from a sika deer (Cervus nippon yesoensis) in Hokkaido, Japan, during the primary culture of sika deer renal cells. This is the first report of isolation of a Megatrypanum trypanosome from Japanese Cervidae. The trypanosome, designated TSD1, was propagated and maintained in Eagle's modified essential medium containing 20% fetal bovine serum with sika deer renal cells as feeder. The TSD1 trypanosome was morphometrically similar to Trypanosoma cervi, which is commonly isolated from American and European deer. PCR analysis with primers for 18S ribosomal DNA and nucleotide sequencing showed that TSD1 is a member of genus Trypanosoma, subgenus Megatrypanum. Phylogenetically TSD1 is closely related to T. theileri, a common trypanosome of cattle, but is distinguishable from T. theileri by some morphometrical and biological features.     

Microvasculature of the rat eye: scanning electron microscopy of vascular corrosion casts

Methylmethacrylate castings of the eye microvasculature were prepared from 10 Spargue-Dawley rats and studied by electron microscopy. The choroidal arterioles are larger in diameter than retinal arterioles, and have a shorter course to choroidal capillaries than retinal arterioles to retinal capillaries. Retinal capillaries are extremely thin in diameter and form a sparse retinal capillary network. The choriocapillaris is large and sinusoid-like, forming a compactly arranged network in the choroid. These differences in the microvasculature between the choroid and retina help explain the differences in ocular hemodynamics; that is, the blood flow in the choroid is faster than that in the retina. Capillaries of the iris show a zigzag configuration, which may be an accommodation for dilation and constriction of the pupil. Capillaries of the ciliary body are of large diameter forming leaf-like configurations, presumably to contribute to the secretion of the aqueous humor. Capillaries of the conjunctiva exhibit a somewhat coiled configuration, the arrangement of which reduce tension of the conjunctiva vessels caused by eyeball movement. Intra-arterial cushions, which control blood flow at the branching sites, are found in both choroidal or retinal arterioles.     

Production of mouse fibroblast growth factor 4 in E. coli and its application for isolation and maintenance of mouse trophoblast stem cells in vitro

Fibroblast growth factor 4 (FGF4) promotes isolation of trophoblast stem (TS) cells from mouse blastocysts and maintenance of TS cells in an undifferentiated state in vitro. To date, commercially available, bacterially expressed human FGF4 (RhFGF4) has been used generally for this purpose. In this study, HismFGF4, a 6x histidine-tagged mouse FGF4, was produced in E. coli and purified using heparin column chromatography. We demonstrated that HismFGF4 (25 ng/ml) more efficiently generates mouse TS cells from a single blastocyst than RhFGF4 (25 ng/ml) and that TS cells isolated and maintained with HismFGF4 retained their ability to differentiate into the trophoblast cell lineage in vitro. In addition, TS cells cultured with HismFGF4 (25 ng/ml) were maintained in an undifferentiated state better than with RhFGF4 (25 ng/ml). To the best of our knowledge, this is the first application of a mouse FGF4 derivative for isolation and maintenance of mouse TS cells.     

Influence of puddling procedures on the quality of rice paddy drainage water

Drainage water quality in rice paddies was strongly influenced by the puddling of soil in the paddy fields by tractors and in response to opening of drainage gates. The concentrations of contaminants in drainage water increased rapidly when the puddling process began and were maintained at high concentrations throughout the puddling period. Moreover, the high concentrations did not decrease immediately after the puddling procedures ceased. Additionally, the ratio of dissolved nitrogen and phosphorous to total nitrogen and total phosphorous increased daily during the last half of the puddling period, due to discharge of chemical fertilizers with the drainage water. Also, the loads of particulate nitrogen and phosphorus discharged during the puddling period were larger than the loads discharge during irrigation. The discharge from paddy fields during puddling also increased the total annual contaminant load.     

Molecular typing of Salmonella enterica serovar Enteritidis isolates from food-producing animals in Japan by multilocus variable-number tandem repeat analysis: evidence of clonal dissemination and replacement

Background

Salmonella enterica serovar Enteritidis is a zoonotic pathogen. Human infections are associated with contaminated eggs and egg products. In Japan, since 1989, the incidence of food-borne disease caused by S. Enteritidis has increased and a pandemic has occurred; however, little is known about changes that occurred before and after this pandemic event in the dominant lineage of isolates from food-producing animals. This study aimed to determine the S. Enteritidis lineages in Japan over the last few decades by using multilocus variable-number tandem repeat analysis (MLVA).

Findings

MLVA was used to analyse 79 S. Enteritidis isolates collected from chickens (n = 63), cattle (n = 12), pigs (n = 2), and goats (n = 2) during 1975–2009. The S. Enteritidis isolates showed 14 different MLVA allele combinations, which were classified into two major clusters (A and C) and a minor cluster (B). All the 62 isolates in cluster A were isolated after 1988, whereas 13 of the 17 isolates belonging to cluster B and C were isolated before 1989.

Conclusions

The MLVA results showed that cluster C was predominant before 1989, and isolates in cluster A disseminated since 1989 and replaced the previous dominant clone, suggesting that isolates of cluster A originated from imported S. Enteritidis infection.     

Direct absorption of acylated anthocyanin in purple-fleshed sweet potato into rats

Absorption of acylated anthocyanins in purple-fleshed sweet potato (Ipomoea batatas cv. Ayamurasaki) in rats was studied to obtain evidence that the acylated anthocyanins themselves could exert a physiological function in vivo. Peonidin 3-caffeoylsophoroside-5-glucoside (Pn 3-Caf*sop-5-glc) in purple-fleshed sweet potato was directly absorbed into rat and present as an intact acylated form in plasma. After oral administration of the purple-fleshed sweet potato anthocyanin (PSA) concentrate containing 38.9 micromol of Pn 3-Caf*sop-5-glc/kg of body weight, Pn 3-Caf*sop-5-glc was detected in the plasma, and the C(max) value and t(max) were estimated as 50.0 +/- 6.8 nmol/Lof plasma and 30 min, respectively. Furthermore, the plasma antioxidant capacity was significantly elevated from 58.0 +/- 12.0 to 89.2 +/- 6.8 micromol of Trolox equivalent/L of plasma 30 min after the administration of the PSA concentrate.