Genetical survey of novel type of Cryptosporidium andersoni in cattle in Japan

Previously, we reported that an isolate of novel type of Cryptosporidium andersoni detected in cattle in Japan contained Type A (identical to C. andersoni reported previously) and Type B (having a thymine nucleotide insertion unlike the Type A) genotypes in the 18S rRNA gene. Here, we conducted an extensive investigation of Cryptosporidium infections in adult cattle in Japan from 2004 to 2007. Consequently, Cryptosporidium sp. were detected in 12 of the 205 cattle examined (5.9%), and partial sequences of the Cryptosporidium oocyst wall protein (COWP) gene in all isolates were identical to those of the previously reported data for C. andersoni whereas two signals were observed in the sequence of the partial 18S rRNA gene in all the isolates. In transmission studies using five of the isolates, they all infected SCID mice. Modified multiplex PCR using DNA of a single oocyst isolated from the infected SCID mice revealed that the partial sequences in the 18S rRNA gene of 40-80% of 10 isolates were identical to the Type A genotype of C. andersoni and those of other samples were identical to the Type B genotype. These results suggested that the C. andersoni novel type is widespread in cattle throughout Japan, and have multiple copies (Types A and B) in the 18S rRNA gene.     

Effects of tall alleles SD1-in and SD1-ja to the dwarfing allele sd1-d originating from ‘Dee-geo-woo-gen’ on yield and related traits on the genetic background of indica IR36 in rice

sd1-d originating from ‘Dee-geo-woo-gen’ has been utilized to develop short-culmed indica varieties adaptable to higher fertilizer-application. Its tall alleles SD1-in and SD1-ja are harbored in indica and japonica subspecies, respectively. The sd1-d of indica IR36 was substituted with SD1-in or SD1-ja by recurrent backcrossing with IR36, and two tall isogenic lines (“5867-36” and “Koshi-36”) were developed. IR36, 5867-36 and Koshi-36 were grown in a paddy field in three years, and yield and related traits were measured, the effects of SD1-in and SD1-ja on yielding ability and related characteristics were examined on the genetic background of IR 36. SD1-in decreased panicle number per m² but increased spikelet number per panicle, ripened-grain percentage and 1000-grain weight, compared with sd1-d, resulting in the increase of yield. The increase of 1000-grain weight by SD1-in, caused by the increases of length, width and thickness of grain, was due to the increases of the length and width of lemma. SD1-ja did not significantly affect yield, mainly because the decrease of panicle number per m² was compensated by the enlarged 1000-grain weight owing to the increase of lemma length. Serious lodging was observed in long-culmed 5867-36, suggesting that sd1-d is indispensable for indica breeding programs.     

Changes in lectin binding patterns of mouse male germ cells (gonocytes) during prespermatogenesis.

The distribution of sugar residues in gonocytes of the differentiating mouse testis was examined by light microscopy using 22 different kinds of lectins. Characteristic binding patterns of sWGA, VVA, and LEA in gonocytes were observed during prespermatogenesis. sWGA preferentially bound to the cytoplasm and plasma membrane of gonocytes on 16.5 days post coitus (dpc). Its reaction decreased thereafter and almost disappeared on 1.5 days post partum (dpp), but reaction reappeared on 4.5 dpp and continued until 6.5 dpp. The VVA reaction was recognized in a few gonocytes on 0.5 dpp, and remained strong until 6.5 dpp. LEA reacted strongly in the plasma membrane and cytoplasm of gonocytes from 0.5 dpp to 6.5 dpp. The present study indicates that sWGA, VVA, and LEA are useful markers for gonocytes, and the appearance or disappearance of sWGA and VVA may be related to the differentiation of gonocytes during prespermatogenesis.     

Establishment and characterization of primary canine hepatocellular carcinoma cell lines producing alpha-fetoprotein

A primary cultured cell line named CHKS was established from a hepatocellular carcinoma (HCC) of a dog showing a high level of serum alpha-fetoprotein (AFP). CHKS secreted a 66 KDD AFP into the growth medium regardless of the presence or absence of fetal bovine serum (FBS). Cloning CHKS with limiting dilution produced 4 clones, CHKS-1, -2, -3, and -4, which secreted 826, 471, 70, and less than 10 ng/ml, respectively, of AFP into the culture medium. In culture, these cell lines were similar in morphology and proliferation pattern to epithelial cells and positive to periodic acid-Schiff (PAS) staining. The presence of mRNA for canine albumin was demonstrated by nested PCR. The doubling times of the clone cell lines were 21, 45, 36, and 35 h, saturation densities 34, 18, 22, and 24 x 10(4)/cm(2), and plating efficiencies 18, 45, 46, and 45%, respectively. Chromosome analysis of these cell lines showed near triploidy. These results show that CHKS and its clones have hepatic cell functions and are useful for carcinogenetic and clinical studies of canine HCC.     

Promotion of liver and kidney carcinogenesis by ethyl tertiary-butyl ether (ETBE) in male Wistar rats

Tumor-promoting effects of ethyl tertiary-butyl ether (ETBE) were investigated in a 2-stage carcinogenesis bioassay with regard to hepatic and renal carcinogenesis in rats. Male 6-week-old Wistar rats were given drinking water containing N-ethyl-N-(2-hydroxyethyl)nitrosamine (EHEN), as an initiator, at a dose of 500 ppm for 2 weeks. Starting one week thereafter, the animals were administered ETBE at dose levels of 0 (control), 100, 300, 500 or 1,000 mg/kg/day by gavage for 19 weeks from week 4 to 22. Necropsy of all rats was performed at week 23, and livers and kidneys were examined histopathologically. Incidences of hepatocellular adenomas, and those of combined hepatocellular adenomas and carcinomas were significantly elevated in rats given 1,000 mg/kg/day ETBE, but not 100‒500 mg/kg/day ETBE, and there was a significant increase in the average numbers of lesions. No significant differences in incidences and average numbers of renal tubule neoplasms were found in rats administered 100‒1,000 mg/kg/day ETBE. However, the average numbers of atypical tubule hyperplasias, considered to be preneoplastic lesions, were significantly increased in rats given ETBE at 1,000 mg/kg/day, but not in rats given 500 mg/kg/day or lower doses. Thus, these results imply that ETBE has hepatic and renal tumor-promoting activities that affect EHEN-induced carcinogenesis in male rats, and the no-observed-effect level is 500 mg/kg/day under the present experimental conditions.     

Three-dimensional structure of Hayabusa samples: origin and evolution of Itokawa regolith

Regolith particles on the asteroid Itokawa were recovered by the Hayabusa mission. Their three-dimensional (3D) structure and other properties, revealed by x-ray microtomography, provide information on regolith formation. Modal abundances of minerals, bulk density (3.4 grams per cubic centimeter), and the 3D textures indicate that the particles represent a mixture of equilibrated and less-equilibrated LL chondrite materials. Evidence for melting was not seen on any of the particles. Some particles have rounded edges. Overall, the particles' size and shape are different from those seen in particles from the lunar regolith. These features suggest that meteoroid impacts on the asteroid surface primarily form much of the regolith particle, and that seismic-induced grain motion in the smooth terrain abrades them over time.     

Real-time observation of adsorbate atom motion above a metal surface

The dynamics of cesium atom motion above the copper(111) surface following electronic excitation with light was studied with femtosecond (10(-15) seconds) time resolution. Unusual changes in the surface electronic structure within 160 femtoseconds after excitation, observed by time-resolved two-photon photoemission spectroscopy, are attributed to atomic motion in a copper-cesium bond-breaking process. Describing the change in energy of the cesium antibonding state with a simple classical model provides information on the mechanical forces acting on cesium atoms that are "turned on" by photoexcitation. Within 160 femtoseconds, the copper-cesium bond extends by 0.35 angstrom from its equilibrium value.     

Rapid enumeration of bacteria grown on surimi-based products by flow cytometry

ABSTRACT: A rapid method to enumerate bacteria adhered on a surimi-based product (kamaboko) by flow cytometry (FCM) is described. To remove Escherichia coli cells from the surface of kamaboko, ultrasonic energy was used. Almost all cells can be removed from kamaboko in 3 min with ultrasonic treatment. Because the sample might contain various non-bacterial particles such as food additives and debris of products, propidium iodide was used to discriminate bacterial cells from non-bacterial particles. Fluorescence scattergrams could distinguish bacteria from the particles, and the FCM method could be used to enumerate bacteria adhered on the surface of kamaboko during storage. Cell numbers determined by FCM paralleled well with those measured using a traditional colony counting method in the range of 10⁴–10⁸ cells/g. The FCM assay could enumerate cells within 1 min and the total assay time, including sample preparation, was less than 30 min.