Polymerase chain reaction (PCR) for the detection of herpesvirus in tortoises

A polymerase chain reaction (PCR)-based method using a herpesvirus consensus primer was assessed for the identification of herpesviral infections in tortoises. A single band of about 230 bp was detected in PCR products from two out of twenty swabs taken from the oral cavity, three out of three paraffin-embedded tissue sections from the liver (two cases) and oral mucosa (one case), and one out of two fresh tissue samples from the oral mucosa. Nucleotide sequencing of these PCR products indicated that the herpesvirus present in these tortoises might belong to the alphaherpesvirinae. PCR using swabs and biopsy tissues was a sensitive and highly specific method for the diagnosis of herpesviral infections in tortoises.     

Isolation of chicken taste buds for real‐time Ca2+ imaging

We isolated chicken taste buds and used a real‐time Ca²⁺ imaging technique to investigate the functions of the taste cells. With RT‐PCR, we found that isolated chicken taste bud‐like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle‐shaped and approximately 61–75 μm wide and 88–98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca²⁺ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L‐glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5′‐monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca²⁺ imaging can be informative.     

Present state of imposex in neptune whelk Neptunea arthritica inhabiting shallow waters around Hokkaido, Japan

ABSTRACT:   The state of imposex in Neptunea arthritica from seven sites along the coast of Hokkaido, Japan was examined in 2002 based on the criteria: (i) relative penis size index (RPSI); (ii) imposex frequency; (iii) stage frequency distribution of imposex in adult and immature whelks; and (iv) sex ratio. RPSI differed from site to site, although values from all sites were low (0.186–5.294). In particular, the RPSI values for four sites were very low (<1.0). In sites where immature whelks were also collected, the frequency of imposex was considerably lower in immature (7.7%–55.6%) than in adult whelks (50%–95.2%) except at one site. The imposex stage frequency distribution also differed among sites, and the trend in the adult whelks corresponded with their RPSI value. Female whelks showing severe imposex (Stage 3 and 3+) were restricted to large individuals.     

An evaluation of one example of biotope roof in Japan: Plant development and invertebrate colonisation after 8 years

Biotope roofs in Japan are usually intensive green roofs that primarily include native plants and food plants for invertebrates and a pond and stones to create a wide range of habitats. The study aimed to evaluate the survivability of planted species, and colonisation by plants and invertebrates on a biotope roof and to suggest an appropriate planting design and maintenance scheme to optimize biodiversity benefits. An intensive green roof (150 m², substrate depth of 50 cm) was installed in 2002 on the ninth storey of a building at Chiba University, Japan. Twelve species of trees, 18 species of shrubs and 8 species of forbs (mainly native species) were planted and volcanic stones were used as mulch and to create habitats for invertebrates. No maintenance and no irrigation were applied for almost 8 years. A limited number of tree species, such as Myrica rubra and Cinnamomum camphora could grew well without maintenance and irrigation at a substrate depth of 50 cm. Overall, shrubs grew successfully and a high density planting seemed effective in increasing wind resistance. Most forbs disappeared, probably because of drought and competition with these colonising plants. Eleven plant species spontaneously colonised resulting in domination by Solidago altissima and Miscanthus sinensis. These two species were too aggressive, and selective weeding is required for species richness. In an invertebrate study, 46 species in 11 orders were observed and the highest number of invertebrate species was observed in the pond and shady areas.     

Computer-Simulated Evaluation of the Hybrid Weakness Gene System as a Means of Preventing Genetic Contamination of Rice Cultivars

Effectivness of the hybrid-weakness caused by a complementary interaction of two dominant genes Hwc₁ and Hwc₂ to suppress the spread of the red-grain gene into ordinary white-grained rice cultivars was investigated using the Monte Carlo computer simulation method. It was shown that the weakness genes, as far as being capable of reducing the fitness of the carrier by 80 percent or more, work quite effectively to suppress the genetic contamination of white-grained cultivars due to inflow of pollen from red-grained cultivars due to inflow of pollen from red-grain gene once the gene has been incorporated in to the population in the form of red-grained progeny produced as a result of the imperfect lethality of the initial hybrid between red-and white-grained cultivars. The weakness genes have little or no effect cultivars. The weakness genes have little or no effect either, when the contamination is initiated by an inflow of seeds or seedlings from ed-grained cultivars in these situations, removal (selection)of red grained plants Is the only countermeasure to exterminate the red-grain gene. The removal of red grained plants need not be intensive if carried out continuously. The spread of the red-grain gene after initial contamination is suppressed if the red-grain gene and weakness genes are linked. The suppressing effect of the linkage, however, is not substantial when the outcrossing rate is around or smaller than 0.01 as can be assumed in most rice cultivars.     

Evaluation of Genetic Diversity of Wild Rice (Oryza rufipogon Griff.) in Myanmar using Simple Sequence Repeats (SSRs)

As a part of an in situ survey of wild rice (Oryza rufipogon Griff.) in Myanmar (Burma), 16 strains of wild rice were collected, and analyzed for allelic diversity over 74 loci with simple sequence repeat (SSR) markers to obtain a basic information for their conservation. Three each of indica and japonica cultivars were added for a comparison. In the six cultivars and 16 strains of wild rice, three to 15 alleles were detected per locus with an average of 7.9. The wild rice revealed a large number of unique alleles throughout their chromosomes with much wider ranges of variation than those detected in the six cultivars of O. sativa L.. The alleles found in the wild rice were classified into those specific to wild rice, common to wild rice and cultivars, and those similar to indica or japonica cultivars. According to the classification, the genotype of each of the 16 strains of wild rice was schematically depicted. The genetic variation among individual strains within a collection site was larger than the variation among the collection sites.     

CTLA-4 control over Foxp3+ regulatory T cell function

Naturally occurring Foxp3+CD4+ regulatory T cells (Tregs) are essential for maintaining immunological self-tolerance and immune homeostasis. Here, we show that a specific deficiency of cytotoxic T lymphocyte antigen 4 (CTLA-4) in Tregs results in spontaneous development of systemic lymphoproliferation, fatal T cell-mediated autoimmune disease, and hyperproduction of immunoglobulin E in mice, and it also produces potent tumor immunity. Treg-specific CTLA-4 deficiency impairs in vivo and in vitro suppressive function of Tregs-in particular, Treg-mediated down-regulation of CD80 and CD86 expression on dendritic cells. Thus, natural Tregs may critically require CTLA-4 to suppress immune responses by affecting the potency of antigen-presenting cells to activate other T cells.     

Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation

Phospholipase A(2)(PLA(2)) enzymes are considered the primary source of arachidonic acid for cyclooxygenase (COX)-mediated biosynthesis of prostaglandins. Here, we show that a distinct pathway exists in brain, where monoacylglycerol lipase (MAGL) hydrolyzes the endocannabinoid 2-arachidonoylglycerol to generate a major arachidonate precursor pool for neuroinflammatory prostaglandins. MAGL-disrupted animals show neuroprotection in a parkinsonian mouse model. These animals are spared the hemorrhaging caused by COX inhibitors in the gut, where prostaglandins are instead regulated by cytosolic PLA(2). These findings identify MAGL as a distinct metabolic node that couples endocannabinoid to prostaglandin signaling networks in the nervous system and suggest that inhibition of this enzyme may be a new and potentially safer way to suppress the proinflammatory cascades that underlie neurodegenerative disorders.