Heavy-ion beam irradiation is an effective technique for reducing major allergens in peanut seeds

Heavy-ion beam irradiation is an effective technique for mutation breeding to produce new cultivars. Heavy-ion beams have high linear energy transfer capable of breaking the double-stranded DNA molecules, thus inducing stable knockout mutants. Here, we report the first application of this technology to produce hypoallergenic peanut lacking major allergens, Ara h 2 and 3, from the Japanese Nakateyutaka variety. After irradiation with either N or C heavy-ion beams at a dose of 100?Gy, seventeen knockout mutants from 11,335 screened M2 seeds were obtained, eight of which lacked either one of the two isoforms of Ara h 2, and the other nine lacked one of the isoforms of Ara h 3. This result indicates that heavy-ion beam irradiation is a powerful means of producing knockout hypoallergenic peanuts.     

Isolation and molecular characterization of Rem2 isoforms in the rainbow trout (Oncorhynchus mykiss): Tissue and central nervous system expression

REM2 is a member of the REM, RAD, and GEM/KIR (RGK) subfamily of RAS superfamily proteins and plays an important role in brain development and function. In this study, two Rem2 isoforms were isolated from the rainbow trout (Oncorhynchus mykiss). The two genes, designated O. mykiss rem2a and rem2b, both encode 304 amino acid proteins with 61% and 62% identities to zebrafish (Danio rerio) Rem2, respectively, and each with 43% identity to mammalian (human) REM2. To our knowledge, this is the first incidence of Rem2 isoforms in a species that are the result of gene duplication. Both isoforms possessed similar tissue expression profiles with the highest levels in the brain. The rem2a gene has significantly higher expression levels than rem2b in all tissues assayed except the brain and head kidney. In the central nervous system, both isoforms showed similar expression levels with the highest levels occurring in the olfactory bulb, cerebrum, and midbrain, though rem2a expression is significantly higher in the spinal cord. Based on known functional roles of Rem2 in synapse development and stem cell proliferation, the characterization of Rem2 in rainbow trout could shed light on its role in adult vertebrate neurogenesis and brain regeneration.     

On the spider genus Amaurobius (Araneae, Amaurobiidae) in India and Nepal

A new species, Amaurobius koponenisp. n., is described from Himachal Pradesh on the basis of a male specimen. A key to all five genera of Amaurobiidae that occur in Asia is provided. Four species from India and Nepal incorrectly assigned to Amaurobius are transferred to three genera of Titanoecidae: Anuvinda milloti (Hubert, 1973), comb. n., Pandava andhraca (Patel & Reddy, 1990), comb. n., Pandava nathabhaii (Patel & Patel, 1975), comb. n., and Titanoeca sharmai (Bastawade, 2008), comb. n.     

Isolation and characterization of tetrahydrofuran-degrading Rhodococcus aetherivorans strain M8

We isolated tetrahydrofuran (THF)-degrading bacteria from waste sludge obtained from a chemical factory in Japan. The isolate designated as strain M8 was identified as Rhodococcus aetherivorans by sequence analysis of its 16S rRNA gene. It grew in a medium containing THF as the sole source of carbon and energy, and its optimal growth pH range and temperature were 6–9 and 37 °C, respectively. Strain M8 grew even in the presence of 35 mM THF. For its growth, this bacterium used 1,4-butanediol and γ-butyrolactone, which are supposed to be metabolites of THF. To elucidate the pathway involved in THF metabolism in strain M8, the resting cell reaction was performed, and the metabolites of THF were analyzed. In the resting cell reaction, 5 mM THF was completely degraded within 5 h. Cells were harvested at 2, 3, and 4 h after the initiation of the reaction; the intermediates accumulated in the cells were extracted using methanol and were derivatized using phenyl boronate. Gas chromatography–mass spectrometry (GC–MS) analysis of the derivatized products showed 4-hydroxybutyrate accumulating in the resting cells. This result suggests that R. aetherivorans strain M8 degrades THF via the oxidation pathway.     

High fat-induced obesity associated with insulin-resistance increases FGF-2 content and causes stromal hyperplasia in rat ventral prostate

Obesity affects sex hormone secretion, which can negatively influence prostatic structure, homeostasis, and disease. This investigation aimed to evaluate the repercussions of obesity induced by a high-fat diet on the rat prostate, with or without treatment with the aromatase inhibitor, Letrozole. Adult Wistar rats were fed a high-fat diet (20% saturated fat, O) for 15?weeks to induce obesity or received a balanced diet (4% fat, C). Then, a group of C and O rats were daily treated with Letrozole (1?mg/kg b.w. per day) for 2?weeks (CL and OL, respectively). Subsequently, ventral prostate was processed for analysis by transmission electron microscopy, immunohistochemistry, and Western blotting. Obesity decreased 70% of the testosterone plasma level. The prostate showed epithelial atrophy and dilated acini in the intermediate portion and epithelial wrinkling in the distal tips. The relative frequency of smooth muscle α-actin in the O group increased by 67%. Ultrastructurally, epithelial cells in obese animals presented altered secretory organelles, lipid droplets, and thicker subjacent fibromuscular layer. Letrozole treatment caused a partial restoration of the prostatic changes caused by obesity. Obesity increased the prostatic content of fibroblast growth factor-2 (FGF-2) by 150%, and Letrozole treatment increased this protein even more in the control and obese groups. This investigation shows that obesity provokes structural and ultrastructural changes in the epithelium of rat prostate; these changes might affect gland homeostasis and physiology. The epithelial and smooth muscle cell hyperplasia and increased FGF-2 expression observed in this experimental model of obesity/insulin-resistance might explain the high frequency of benign prostatic hyperplasia in insulin-resistant men.     

Solution structure and stability of the DNA undecamer duplexes containing oxanine mismatch

Solution structures of DNA duplexes containing oxanine (Oxa, O) opposite a cytosine (O:C duplex) and opposite a thymine (O:T duplex) have been solved by the combined use of (1)H NMR and restrained molecular dynamics calculation. One mismatch pair was introduced into the center of the 11-mer duplex of [d(GTGACO(6)CACTG)/d(CAGTGX(17)GTCAC), X = C or T]. (1)H NMR chemical shifts and nuclear Overhauser enhancement (NOE) intensities indicate that both the duplexes adopt an overall right-handed B-type conformation. Exchangeable resonances of C(17) 4-amino proton of the O:C duplex and of T(17) imino proton of O:T duplex showed unusual chemical shifts, and disappeared with temperature increasing up to 30 °C, although the melting temperatures were >50 °C. The O:C mismatch takes a wobble geometry with positive shear parameter where the Oxa ring shifted toward the major groove and the paired C(17) toward the minor groove, while, in the O:T mismatch pair with the negative shear, the Oxa ring slightly shifted toward the minor groove and the paired T(17) toward the major groove. The Oxa mismatch pairs can be wobbled largely because of no hydrogen bond to the O1 position of the Oxa base, and may occupy positions in the strands that optimize the stacking with adjacent bases.     

Coexpression of three middle wavelength-absorbing visual pigments in sexually dimorphic photoreceptors of the butterfly Colias erate

The tiered ommatidia of the Eastern Pale Clouded yellow butterfly, Colias erate, contain nine photoreceptor cells, four of which contribute their rhabdomeral microvilli to the distal tier of the rhabdom. We analyzed the visual pigments and spectral sensitivities of these distal photoreceptors in both sexes of Colias erate. A subset of photoreceptor cells expresses a newly discovered middle wavelength-absorbing opsin, C olias e rate Blue (CeB), in addition to two previously described middle wavelength-absorbing opsins, CeV1 and CeV2. The other photoreceptors either coexpress CeV1 and CeV2, or exclusively express a short wavelength-absorbing opsin, CeUV, or a long wavelength-absorbing opsin, CeL. Males and females have the same visual pigment expression patterns, but the photoreceptor spectral sensitivities are sexually dimorphic. The photoreceptors coexpressing three middle wavelength-absorbing opsins are broad-blue receptors in males, but in females they are narrow-blue receptors. Those with CeV1 and CeV2 are violet receptors in females, while they are shouldered-blue receptors in males. The sexual dimorphism in spectral sensitivity is caused by a sex-specific distribution of fluorescent pigment that functions as a spectral filter.     

Essential Roles of Gangliosides in the Formation and Maintenance of Membrane Microdomains in Brain Tissues

Gangliosides are considered to be involved in the maintenance and repair of nervous tissues. Recently, novel roles of gangliosides in the regulation of complement system were reported. Here we summarized roles of gangliosides in the formation and maintenance of membrane microdomains in brain tissues by comparing complement activation, inflammatory reaction and disruption of glycolipid-enriched microdomain (GEM)/rafts among several mutant mice of ganglioside synthases. Depending on the defects in ganglioside compositions, corresponding up-regulation of complement-related genes, proliferation of astrocytes and infiltration of microglia were found with gradual severity. Immunoblotting of fractions separated by sucrose density gradient ultracentrifugation revealed that DAF and NCAM having GPI-anchors tended to disappear from the raft fraction with intensities of DKO > GM2/GD2 synthase KO > GD3 synthase KO > WT. The lipid raft markers tended to disperse from the raft fractions with similar intensities. Phospholipids and cholesterol also tended to decrease in GEM/rafts in GM2/GD2 synthase KO and DKO, although total amounts were almost equivalent. All these results indicate that GEM/rafts architecture is destroyed by ganglioside deficiency with gradual intensity depending on the degree of defects of their compositions. Implication of inflammation caused by deficiency of gangliosides in various neurodegenerative diseases was discussed.