Molecular characterisation of <Emphasis Type="Italic">Turnip mosaic virus</Emphasis> isolates from Brassicaceae weeds

Eight provinces of Iran were surveyed during 2003–2008 to find Brassicaceae reservoir weed hosts of Turnip mosaic virus (TuMV). A total of 532 weed samples were collected from plants with virus-like symptoms. The samples were tested for the presence of TuMV by enzyme-linked immunosorbent assay using specific antibodies. Among those tested, 340 samples (64%) were found to be infected with TuMV. Rapistrum rugosum, Sisymberium loeselii, S. irio and Hirschfeldia incana were identified as the Brassicaceae weed hosts of TuMV, and the former two plant species were found to be the most important weed hosts for the virus in Iran. The full-length sequences of the genomic RNAs of IRN TRa6 and IRN SS5 isolates from R. rugosum and S. loeselii were determined. No evidence of recombination was found in both isolates using different recombination-detecting programmes. Phylogenetic analyses of the weed isolates with representative isolates from the world showed that the IRN TRa6 and IRN SS5 isolates fell into an ancestral basal-Brassica group. This study shows for the first time the wide distribution and phylogenetic relationships of TuMV from weeds in the mid-Eurasia of Iran.     

Postnatal development of the mouse volatile papilla taste bud cells

In the present study, we examined specific markers for taste bud cells in the mouse and the postnatal development of volatile papilla taste bud cells in ddY mice. We examined the immunoreactivity of 4 types of carbonic anhydrase isoenzymes, CA I, CA II, CA III and CA VI, as specific markers for taste bud cells, and K8.13 cytokeratin antibody as a specific marker for the lingual epithelial cells. Of the carbonic anhydrase isoenzymes, only CA III immunoreactivity was clearly detected in the spindle shaped gustatory cells. CA VI immunoreactivity was detectable in suspentacular cells. CA I and CA II antibodies did not recognize any taste bud cell specifically. K8.13 cytokeratin immunoreactivity was detected in the lingual epithelial cells, but not in taste bud cells. At 7 days after birth, the suckling phase, very small taste buds developed from the anaplastic gustatory cells. At 14 days after birth, the taste buds showed larger size than those at 7 days after birth. At 21 days birth, after the weaning phase, taste bud structure approximated the mature structure. These results demonstrate the specificity of anti-CA III and anti-CA VI for gustatory cells and suspentacular cells, respectively. These markers should be useful for an analysis of taste bud development in mice.     

Hepatic lymphangiomatosis in a young dog

A 9-month-old intact male American cocker spaniel was referred because of hepatomegaly and ascites. Ultrasonographic evaluation of the liver revealed congestion and increased parenchymal echogenicity with focal and more hyperechoic nodules. Histopathology of the hepatic lesion revealed diffuse, ill-defined vascular proliferation. A single layer of endothelial cells, which showed signs of minimal cellular atypia, lined the irregular vessels. On immunohistochemistry, the proliferative endothelial cells lining the irregular vessels were positive for an antiserum to factor VIII related antigen. Based on these findings, the dog was diagnosed with hepatic lymphangiomatosis.     

Effects of chemical ischemia on purine nucleotides,free radical generation,lipids peroxidation and intracellular calcium levels in C2C12 myotube derived from mouse myocytes

To elucidate the mechanisms of ischemia-mediated myopathy using in vitro model, changes of purine nucleotides, membrane lipid peroxidation(TBARS), intracellular calcium ([Ca2+]i)levels, generation of free radicals, and deoxyribonucleic acid (DNA) fragmentation were examined in mouse-derived C2C12 myotubes under the condition with an inhibition of glycolytic and oxidative metabolism as the ischemic condition. In purine nucleotides, intracellular adenosine triphosphate (ATP) and guanosine triphosphate (GTP) concentrations rapidly and significantly decreased after the treatment with ischemia. No remarkable differences were observed in other purine nucleotides, with the exception of inosine monophosphate (IMP) and extracellular hypoxanthine levels, both of which increased significantly during the ischemia. The lactate dehydrogenase activity in culture supernatant of C2C12 myotubes increased significantly from 2 to 4 hr after the ischemia. On the generation of free radicals, no spectrum was detected in supernatants throughout the observation period, whereas supernatant TBARS concentration increased rapidly and significantly after the ischemia. The relative intensity of [Ca2+]i significantly increased after the ischemia. On the fragmented deoxyribonucleic acid(DNA), no TUNEL positive cells was detected in C2C12 myotubes after 1 hr of the ischemia, however the positive cell percentage subsequently increased. From these results, it was suggested that the ischemic condition induced changes of membrane permeability and increase of [Ca2+]i, both of which lead to cell membrane damage, although a free radical generation was not detected. The ischemic condition also induced the release of substrate hypoxanthine for free radical generation and might initiate the apoptotic pathway in C2C12 myotubes.     

Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.     

Validity of fluctuating asymmetry as a gauge of genetic stress in ayu stocks

An increasing demand for stocking of hatchery-reared ayu, Plecoglossus altivelis, has aroused concerns about their genetic status which has an effect on their post-stocking performance and the conservation of native biodiversity. Instead of costly analysis, fluctuating asymmetry (FA) was examined to test its suitability as an indirect measure of genetic diversity. Two bilateral traits, pectoral and pelvic fin rays, were targeted, and two wild and four hatchery populations of ayu were sampled for comparison. Frequency of FA in soft-rays was not similar between specimens in both pelvic and pectoral fins. The wild populations did not show FA incidences lower than the hatchery populations, suggesting a considerable degree of environmental stress operating in the wild populations. Comparison between the hatchery populations showed overall increasing FA incidence with the decrease in genetic diversity, although the correlation was not complete in detail. In conclusion, FA can be used as a convenient tool to compare genetic status, but quantified evaluation is rather limited to monitoring changes in the genetic variation within a hatchery population.     

Complete mitochondrial DNA sequence of ayu Plecoglossus altivelis

SUMMARY: We determined the complete nucleotide sequence of the mitochondrial genome for ayu, Plecoglossus altivelis . Two large DNA fragments covering the entire genome were amplified using a long polymerase chain reaction (PCR) technique, and the products subsequently used as templates for PCR with 57 fish-versatile and five species-specific primers that amplify contiguous, overlapping segments of the entire genome. Direct sequencing of the PCR products demonstrated that the genome (16 537 bp) contained the same 37 mitochondrial genes (two ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes) as those found in other vertebrates, with the gene order identical to that in typical vertebrates. A major non-coding region between the tRNA^Pro and tRNA^Phe genes (857 bp) was considered to be the control region (D-loop), as it has several conservative blocks that are characteristic to this region.