Tumor necrosis factor-alpha induces apoptosis in cultured porcine luteal cells

Some studies in mammalian species recently demonstrated that tumor necrosis factor (TNF)-alpha plays an important role for corpus luteum (CL) function by way of apoptosis during the estrous cycle. The objectives of this study were to clarify the induction of apoptosis in cultured porcine luteal cells by TNF-alpha treatment. Luteal cells prepared from porcine ovaries collected from crossbred mature gilts on Days 10-14 of the estrous cycle were isolated and examined as follows: 1) Flow cytometric analysis was carried out to determine apoptosis in cultured luteal cells. 2) Single-cell gel electrophoresis (comet assay) was performed to investigate apoptotic DNA fragmentation in luteal cells. The results of the flow cytometric analysis and comet assay demonstrated coincidentally that TNF-alpha induces DNA fragmentation in luteal cells causing apoptosis. These results revealed that TNF-alpha is an inducing factor of apoptosis in luteal cells.     

Cellular localization of IL-18 and IL-18 receptor in pig anterior pituitary gland

Pro-inflammatory cytokine interleukin 18 (IL-18) has been proposed to have a role in modulating immuno-endocrine functions. Our previous study showed that IL-18 and IL-18 receptor (IL-18R) colocalized in somatotrophs of the bovine anterior pituitary gland, and the possibility that IL-18 acts on somatotrophs as an autocrine factor. In the present study, we investigated the localization of IL-18 and IL-18R in the pig anterior pituitary gland. RT-PCR analysis showed the expression of IL-18 and IL-18R mRNAin the pig anterior pituitary gland. Immunohistochemistry of IL-18 and specific hormones revealed the presence of IL-18 in somatotrophs, mammotrophs, thyrotrophs and gonadotrophs. IL-18R was localized in somatotrophs and thyrotrophs. Furthermore, the somatotrophs immunoreactive for IL-18 did not contain IL-18R. Thus, IL-18R and IL-18 were not colocalized in an identical somatotroph. These findings suggest that the localization of IL-18 in pig somatotrophs is different from that in bovine somatotrophs, although IL-18 closely associates with somatotrophs in the anterior pituitary glands in both species.     

Genetic differentiation of the bronze featherback Notopterus notopterus between Mekong River and Tonle Sap Lake populations by mitochondrial DNA analysis

ABSTRACT:   Although the bronze featherback Notopterus notopterus is an important fish in South-East Asia, the population structure has not been investigated. In this study, genetic diversity and population structure of the bronze featherback were examined using nucleotide sequence analysis of the mitochondrial DNA control region for 332 fish collected from Tonle Sap Lake and the Mekong River in Cambodia. The average nucleotide diversity (π) of all samples was 0.033, and the Mekong River samples had higher nucleotide diversity (0.034) than Tonle Sap Lake samples (0.028). The nucleotide diversity between the lake and river samples varied from 0.029 to 0.037. The genetic differentiation between the river and lake populations was also supported by the pairwise F -statistic values and hierarchical analysis of molecular variance, indicating that the Tonle Sap Lake population is genetically isolated from the population in main stream of the Mekong River.     

Genetic diversity of Central Asian and north Caucasian Aegilops species as revealed by RAPD markers

RAPD analysis of 112 accessions of Aegilops tauschii Coss. (genome DD), Ae. cylindrica Host (CCDD), Ae. crassa Boiss. (DDMM), Ae. biuncialis Vis. (UUMM) and Ae. triuncialis L. (UUCC) collected in the Central Asia and north Caucasia was conducted. Aegilops accessions were divided into two major groups, corresponding to the D genome species and the U genome species. These groups were also separated into sub-groups according to species, except for the Ae. tauschii-cylindrica complex of accessions from Central Asia. Aegilops tauschii from north Caucasia was divided into two varietal groups, tauschii and meyeri. The Central Asian accessions of Aegilops species were more diverse than the accessions from north Caucasia. Aegilops tauschii and Ae. cylindrica accessions from north Caucasia were genetically uniform. Associations between altitudal variation of Aegilops species and variability of RAPD markers were not found.     

Genetic variation of high-molecular-weight glutenin subunit composition in Asian wheat

To clarify the genetic properties of the HMW glutenin subunit composition of Asian endemic wheats, SDS–PAGE analysis was conducted using 1,139 bread wheat accessions that were originally collected in Asia. The samples were divided into six regional groups, Western Asia, Caucasia, Central Asia, Afghanistan, Southern Asia, and Eastern Asia. The genotype Glu-A1c, Glu-B1b, and Glu-D1a encoding subunits null, 7+8, and 2+12 had an overall frequency of 55.2%. Thus, we conclude that it is the typical genotype of the HMW glutenin subunits that characterize Asian endemic wheat. The frequency of the typical Asian genotype was relatively high in the central belt of Asia (Western Asia, Afghanistan, and Eastern Asia) and low in the marginal regions (Caucasia, Central Asia, and Southern Asia). In Southern Asia, the frequency of Glu-B1i, which encodes subunit 17+18, was the highest at the Glu-B1 locus. In Caucasia and Central Asia, the frequency of Glu-D1d, which encodes subunit 5+10 (which is considered to be the most useful for making bread), was high. The level of genetic variation, as estimated using the frequencies of the various alleles, was relatively low in the central belt of Asia and high in the marginal regions. Among the three Glu-1 loci, the highest number of alleles was detected at the Glu-D1 locus. This result was caused by the presence of rare Asian specific alleles at the Glu-D1 locus, in which a newly found allele, Glu-D1bs, encoding subunit 2.1+12 was included.     

Analysis of acrylamide in green tea by gas chromatography-mass spectrometry

Optimization of the solid-phase extraction cleanup procedure enabled the GC-MS analysis of acrylamide in tea samples without the interference of bromination by tea catechins. Although polyvinylpolypyrrolidone (PVPP) is available for removing tea catechins from tea extract, the peaks derived from PVPP had the same retention time as brominated acrylamide in mass chromatograms obtained by GC-MS. A considerable amount of acrylamide was formed at roasting temperatures of > or =120 degrees C; the highest acrylamide level was observed when tea samples were roasted at 180 degrees C for 10 min. Higher temperatures and longer processing times caused a decrease in the acrylamide content. Furthermore, an analysis of 82 tea samples showed that rather than the reducing sugar content, the asparagine content in tea leaves was a significant factor related to acrylamide formation in roasted products. The acrylamide level in roasted tea products was controlled by asparagine in the presence of reducing sugars.     

Anthracnose of belmore sentry palm (<Emphasis Type="Italic">Howea belmoreana</Emphasis> Becc.) caused by <Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis> (Penzig) Penzig et Saccardo

Severe leaf spots were found on the ornamental plant, belmore sentry palm (Howea belmoreana), grown in pots in a greenhouse in Ibusuki, Kagoshima, Japan in 2006. The isolated fungus caused the same symptoms after artificial inoculation and was identified morphologically and molecularly as Colletotrichum gloeosporioides. In this first report of the disease, the name anthracnose of belmore sentry palm (kenchayashi-tanso-byo in Japanese) is proposed.     

Production efficiency and telomere length of the cloned pigs following serial somatic cell nuclear transfer

The aim of the present study was to examine the production efficiency of cloned pigs by serial somatic cell nuclear transfer (SCNT) and to ascertain any changes in the telomere lengths of multiple generations of pigs. Using fetal fibroblasts as the starting nuclear donor cells, porcine salivary gland progenitor cells were collected from the resultant first-generation cloned pigs to successively produce second- and third-generation clones, with no significant differences in production efficiency, which ranged from 1.4% (2/140) to 3.3% (13/391) among the 3 generations. The average telomere lengths (terminal restriction fragment values) for the first, second and third generation clones were 16.3, 18.1 and 20.5 kb, respectively, and were comparable to those in age-matched controls. These findings suggest that third-generation cloned pigs can be produced by serial somatic cell cloning without compromising production efficiency and that the telomere lengths of cloned pigs from the first to third generations are normal.     

An enzyme-linked immunosorbent assay using canine coronavirus-infected CRFK cells as antigen for detection of anti-coronavirus antibody in cat

From the reasons that canine coronavirus (CCV) grows more efficiently than feline coronavirus in a cell culture and they are mutually related in their antigenicities, an enzyme-linked immunosorbent assay (ELISA) using CCV-infected feline kidney (CRFK) cells as substrate antigens was developed for detection of anti-coronavirus antibodies in cats. It was indispensable for generating coronavirus-specific ELISA antibody activities that the sample was applied to the mock-infected, normal CRFK cells in parallel with the CCV-infected cells and then the optical density values given by the mock-infected cell antigen were subtracted from those given by the virus-infected cell antigen. On the basis of ELISA antibody titers obtained in sera from the cats experimentally infected with CCV and from the spontaneous feline infectious peritonitis (FIP) cases, the ELISA described in the present study was found to be applicable as a simple and easy serologic test which was able to detect anti-coronavirus antibodies as efficiently as the indirect immunofluorescence assay with homologous FIP virus.     

A clinical case of severe anemia in a sheep coinfected with Mycoplasma ovis and 'Candidatus Mycoplasma haemovis' in Hokkaido, Japan

A 2-year-old East Friesian sheep imported from Australia exhibited severe anemia after contagious pustular dermatitis in Hokkaido, Japan. Hemoplasma infection was confirmed in blood smears. Both Mycoplasma ovis and 'Candidatus Mycoplasma haemovis' were detected by PCR and sequence analyses. In the epidemiological analysis, dual pathogens were detected in 6 of 12 (50.0%) sheep imported from Australia with the infected ewe at the same time, 1 of 5 (20.0%) sheep introduced from a domestic farm in Hokkaido, and in 1 of 16 (6.3%) sheep from an epidemiologically unrelated ranch. It is the first clinical case of sheep to confirm coinfection of these pathogens in Japan.