Low expression of cyclin D2 in G2/M-arrested and transformed proliferating Balb/3T3 cells

At present, the effect of cyclin D2 implicated in cell cycle regulation, differentiation, and oncogenic transformation is not fully confirmed. To better elucidate the role of cyclin D2 in controling the cell proliferation, cyclin D2 expression level was determined at the early initiation and promotion stages during the in vitro two-stage transformation process of Balb/3T3 A31 cells. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced G2/M-arrested cells expressed low level of cyclin D2 mRNA, while the contact-inhibited nonproliferating cells expressed high level of cyclin D2 mRNA. In the transformed proliferating cells at the promotion stage, cyclin D2 mRNA was not expressed. These data suggest that cyclin D2 expression may be associated with the type of growth arrest and nonproliferating state, but not with the cell proliferation and transformation.     

Immunohistochemistry and polymerase chain reaction for the detection of Lawsonia intracellularis in porcine intestinal tissues with proliferative enteropathy

Detection method of Lawsonia intracellularis was studied in formalin-fixed paraffin-embedded intestinal tissues from 5 naturally infected pigs by immunohistochemistry with a monoclonal antibody against outer membrane protein of L. intracellularis. Warthin-Starry silver stain revealed clusters of argyrophilic, slightly curved rod-shaped organisms in the apical cytoplasm of enterocytes. Immunohistochemical staining with a L. intracellularis-specific monoclonal antibody confirmed the presence of the organism in the apical cytoplasm of hyperplastic enterocytes. The presence of L. intracellularis in the ileum of pig with proliferative enteropathy was confirmed by polymerase chain reaction (PCR) further on the basis of amplification of 319 base pair products specific for porcine L. intracellularis chromosomal DNA. Immunohistochemistry and PCR may be a complementary method to confirm the diagnosis of L. intracellularis infection in pigs.     

Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing

The present study was to construct a parentage testing system for Thoroughbred (TB) horse. A total number of 1,285 TB horse samples including 962 foals for parentage testing, 9 sires and 314 dams for individual identification were genotyped. Genomic DNA was extracted from 5 hair roots and genotyped by using 14 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS3, HMS6, HMS7, HTG4, HTG10, LEX3 and VHL20). This method consisted of multiplexing PCR procedure and showed reasonable amplification of all PCR products. Genotypes were determined by genetic analyzer. The number of alleles per locus varied from 3 to 9 with a mean value of 6.36 in TB horse. The expected heterozygosity was ranged from 0.548 to 0.831 (mean 0.699), and the total exclusion probability of 14 microstellite loci was 0.9998. Of the 14 markers, ASB2, ASB17, ASB23, HMS7 and HTG10 loci have relatively high PIC value (> 0.7). Of the 962 foals, 960 foals were qualified by compatibility according to the Mendelism. These results suggest that the DNA typing method has high potential for parentage verification and individual identification of TB horses.     

Anti‐microbial Susceptibility for east1+Escherichia coli Isolated from Diarrheic Pigs in Korea

The in vitro susceptibilities of 128 isolates of east1⁺Escherichia coli from pre‐weaned and post‐weaned pigs with diarrhoea were tested with nine commonly used anti‐microbial agents by an agar dilution minimal inhibitory concentration (MIC) procedure according to National Committee for Clinical Laboratory Standards guidelines. For the isolates from pre‐weaned and post‐weaned pigs, most of them were susceptible to low concentrations (MIC₉₀) of tetracycline (4 and 2 μg/ml), ceftiofur (2 and 2 μg/ml), and colistin (4 and 2 μg/ml). Marked resistance was found in others.     

2-fluoroabscisic acid analogues: their synthesis and biological activities

Fluorine was introduced into the 2-position of the side chain of abscisic acid (ABA) analogues by Wittig reaction of alpha-ionone derivatives with ethyl triethylphosphono-2-fluoroacetate. The effects of the fluorinated analogues were evaluated on inhibition of cress seed germination and inhibition of gibberellin-inducible alpha-amylase induction in embryoless barley half-seeds. (2E, 4E)-2-Fluoro-5-(1'-hydroxy-2',6', 6'-trimethyl-2'-cyclohexen-1'-yl)-3-methyl-2,4-pentadienoic acid (5b) showed potent inhibitory activity at the same level as ABA in the cress seed germination test, and 5b also inhibited gibberellin-inducible alpha-amylase induction at 4 x 10(-)(6), 3 times the concentration of ABA (1 x 10(-)(6)) for 50% inhibition of alpha-amylase production. 5b also showed dehydrin induction activity. These results indicate that fluorinated ABA analogues mimic ABA action and can be a lead for a plant growth regulator which regulates plant growth or protects plants from environmental stresses.     

Hydrothermal Assessment of Temporal Variability in Seedbed Microclimate

The microclimatic requirements for successful seedling establishment are much more restrictive than those required for adult plant survival. The purpose of the current study was to use hydrothermal germination models and a soil energy and water flux model to evaluate intra- and interannual variability in seedbed microclimate relative to potential germination response of six perennial grasses and cheatgrass. We used a 44-yr weather record to parameterize a seedbed microclimate model for estimation of hourly temperature and moisture at seeding depth for a sandy loam soil type at the Orchard Field Test Site in southwestern Ada County, Idaho. Hydrothermal germination response was measured in the laboratory for two seed lots of cheatgrass (Bromus tectorum L.), four seed lots of bluebunch wheatgrass (Pseudoroegneria spicata [Pursh] Löve), three seed lots of bottlebrush squirreltail (Elymus elymoides [Raf] Swezey), and one seed lot each of Sandberg bluegrass (Poa secunda J. Presl.), big squirreltail (Elymus multisetus [J.G. Smith] M.E. Jones), thickspike wheatgrass (Elymus lanceolatus [Scribn. And J.G. Smith] Gould) and Idaho fescue (Festuca idahoensis Elmer). Germination response models were developed to estimate potential germination rate for 13 subpopulations of each seed lot for every hour of the 44-yr simulation. Seedbed microclimate was assessed seasonally and for each day, month, and year, and germination rate-sum estimates integrated for a numerical index of relative site favorability for germination for each time period. The rate-sum favorability index showed a consistent pattern among seed lots for different years, and provides a relatively sensitive indicator of annual and seasonal variability in seedbed microclimate. This index could be used with field data to define minimum weather thresholds for successful establishment of alternative plant materials, in conjunction with weather forecast models for making restoration and fire-rehabilitation management decisions in the fall season, for evaluation of potential climate-change impacts on plant community trajectories, and in optimization schemes for selecting among alternative restoration/rehabilitation management scenarios.     

Epizootiologic survey for Babesia microti among small wild mammals in northeastern Eurasia and a geographic diversity in the beta-tubulin gene sequences

We previously reported that small wild rodents in Japan harbor two types of novel Babesia microti-like parasites (designated as Hobetsu and Kobe types), but not the type commonly found in the northeastern United States (U.S. type) where human babesiosis is endemic. To determine whether these new types of parasites are distributed in places surrounding Japan, an epizootiologic survey was undertaken in three geographically distant areas in northeastern Eurasia; South Korea, Vladivostok in Russia, and Xinjiang in China. Blood samples were collected from a total of 387 animals comprising 24 species. DNAs extracted from the samples were tested by nested PCR targeting babesial nuclear small-subunit rRNA gene (rDNA), which revealed that small rodents harboring B. microti exist in all three survey areas. Sequence analysis showed that all PCR-positive samples had rDNA sequences virtually identical to that of U.S.-type B. microti. However, when beta-tubulin gene sequences were compared, evident geographic variations were seen. By use of primers specific for each of the beta-tubulin genes of Kobe-, Hobetsu-, and U.S.-type parasites, a type-specific PCR was developed. Parasite with Hobetsu- or Kobe-type sequence was not detected from any of the three survey areas. These findings suggest that U.S.-type B. microti is widely distributed among small wild mammals in temperate zones of not only North America, but also Eurasia, whereas that Hobetsu- and Kobe-type parasites may be uniquely distributed in Japan.