Infrared spectroscopic evidence for protonated water clusters forming nanoscale cages

Size-dependent development of the hydrogen bond network structure in large sized clusters of protonated water, H+(H2O)n (n = 4 to 27), was probed by infrared spectroscopy of OH stretches. Spectral changes with cluster size demonstrate that the chain structures at small sizes (n less, similar 10) develop into two-dimensional net structures (approximately 10 < n < 21), and then into nanometer-scaled cages (n >/= 21).     

Genetically modified rapeseed oil containing cis-9,trans-11,cis-13-octadecatrienoic acid affects body fat mass and lipid metabolism in mice

Punicic acid, one of the conjugated linolenic acid (CLN) isomers, exerts a body-fat reducing effect. Although punicic acid is found in pomegranate and Tricosanthes kirilowii seeds, the amount of this fatty acid is very low in nature. The goal of this study was to produce a transgenic oil containing punicic acid. A cDNA encoding conjugase that converts linoleic acid to punicic acid was isolated from T. kirilowii, and the plant expression vector, pKN-TkFac, was generated. The pKN-TkFac was introduced into Brassica napus by Agrobacterium-mediated transformation. As a result, a genetically modified rapeseed oil (GMRO) containing punicic acid was obtained, although its proportion to the total fatty acids was very low (approximately 2.5%). The effects of feeding GMRO in ICR CD-1 male mice were then examined. Wild-type rapeseed (B. napus) oil (RSO) containing no CLN was used as a control oil. For reference oils, RSO-based blended oils were prepared by mixing with different levels of pomegranate oil (PO), either 2.5% (RSO + PO) or 5.0% (RSO + 2PO) punicic acid. Mice were fed purified diets containing 10% of either RSO, RSO + PO, RSO + 2PO, or GMRO for 4 weeks, and dietary PO dose-dependently reduced perirenal adipose tissue weight with a significant difference between the RSO group and the RSO + 2PO group. GMRO, as compared to RSO, lowered the adipose tissue weight to the levels observed with RSO + 2PO. The liver triglyceride level of the RSO + 2PO and GMRO groups but not that of the RSO + PO group was lower than that of the RSO group. The RSO + 2PO and GMRO groups, but not the RSO + PO group, had increased carnitine-palmitoyltransferase activity in the liver and brown adipose tissue. These results showed that dietary GMRO, even at a dietary punicic acid level as low as 0.25 wt % of diet, reduced body fat mass and altered liver lipid metabolism in mice and was more effective than an equal amount of punicic acid from PO.     

Differences in dry matter production,grain production,and photosynthetic rate in barley cultivars under long-term salinity

Soil salinity is a major environmental stress causing significant loss of crop productivity. Barley (Hordeum vulgare L.) is one of the few field crops that can grow in salt-affected fields and varietal differences in productivity under salinity conditions were known. To clarify the trait most responsible for grain production under salt stress, barley cultivars that were salt tolerant (OUE812) or salt sensitive (OUC613) were grown from seedling to harvest stage in vermiculite containing various concentrations of NaCl. Dry weight of aboveground parts and grain weight decreased significantly with increasing NaCl concentration. The dry weight of the aboveground parts and grain weight decreased more significantly in OUC613 than in OUE812 for plants treated with 150 mM and 200 mM NaCl. A marked reduction in ripening percentage caused significantly decreased grain production in OUC613 as compared with OUE812. In plants treated with 200 mM NaCl, the photosynthetic rate decreased three weeks after starting the NaCl treatment, but a significant difference between cultivars in photosynthetic rate did not appear until seven weeks of NaCl treatment. OUE812 kept a higher photosynthetic rate during ripening than did OUC613 and dry matter production during the period from ripening to harvest was significantly larger in OUE812 than in OUC613. Keeping a higher photosynthetic rate might have contributed to higher grain production in OUE812. Higher ripening percentage and higher rate of photosynthesis during ripening might be target traits in breeding to improve the tolerance of barley to long-term salt stress.     

Effects of sedimentation on an endangered benthic fish,Cobitis shikokuensis: is sediment‐free habitat a requirement or a preference?

A negative correlation between the degree of fine sediment deposition and the abundance of stream benthic fishes has often been reported, although the causal mechanisms for this effect are not fully understood. To better understand the influence of sedimentation, it is important to clarify whether it alters the habitat required by fish, or merely preferred by them. We conducted two enclosure experiments in the field to examine (i) whether an endangered benthic fish, Cobitis shikokuensis, prefers sediment‐free substrate and (ii) whether fine sediment deposition has negative effects on the physiological condition of C. shikokuensis. The first experiment, which used three types of substrates, ‘cobble’, ‘pebble’ and ‘mixture of fine sediment and pebbles’, showed that C. shikokuensis avoided the mixture. The second experiment, which compared the physiological condition of fish reared in enclosures with either a pebble or mixture substrate, showed that the condition of the fish in the latter group declined more quickly. These results suggest that, for C. shikokuensis, sediment‐free substrate is a habitat requirement, not merely a preference, and that excessive input of fine sediment due to human activities can cause a decline in the population of this species.     

Bovine oocytes in secondary follicles grow in medium containing bovine plasma after vitrification

There has been no culture system that supports the growth of bovine oocytes for more than 2 weeks. In the present study, bovine secondary follicles were cultured for 4 weeks, and the effects of supplemented protein components and FSH in the culture medium on the growth of the oocytes were examined. The effect of vitrification of secondary follicles on the subsequent oocyte growth was also examined. Secondary follicles (150 to 200 μm in diameter) containing growing oocytes (approximately 60 μm in diameter) were dissected from ovaries and cultured in a medium supplemented with FSH (0, 25 or 50 ng/ml) and one of the following four kinds of protein components: bovine serum albumin (BSA), bovine plasma (BPL), fetal calf serum (FCS) and bovine follicular fluid (BFF). In BSA- and BPL-supplemented media with 0 or 25 ng/ml FSH, more than 50% of follicles showed no degenerative signs during culture, and oocytes significantly increased in size after 4 weeks (P<0.05). Higher percentages of granulosa cell-enclosed oocytes were recovered from the follicles cultured in BPL-supplemented media with 0 and 25 ng/ml FSH, and the oocytes grew to 90 μm or more in diameter. In FCS- and BFF-supplemented media, FSH increased the numbers of degenerating follicles. Next, vitrified-warmed secondary follicles were cultured in BPL-supplemented medium. One third of the follicles showed no degenerative signs, and the oocytes increased in diameter to 88.8 ± 3.1 μm after 4 weeks of culture. These results suggest that a BPL-supplemented medium supports oocyte growth in bovine secondary follicles for 4 weeks, even after vitrification and warming of the follicles.     

One-step Generation of Phenotype-expressing Triple-knockout Mice with Heritable Mutated Alleles by the CRISPR/Cas9 System

Triple-knockout mice generated by the one-step CRISPR/Cas9 system were examined for the effects of multiple gene modifications on each phenotype and individual gene function. Sixty embryos were transferred, and 9 pups were obtained; all 9 pups had mutations on 3 loci, and 7 pups showed mutations in all-alleles. F0 mice showed knockout phenotypes or no protein expression of target genes simultaneously, and these mutations were normally inherited in the next generation.     

Red:far-red light ratio and far-red light integral promote or retard growth and flowering in Eustoma grandiflorum (Raf.) Shinn

Night break treatment was applied to Eustoma grandiflorum ‘Nail Peach Neo’ using light sources with different red (R: 660 ± 30 nm): far-red (FR: 730 ± 30 nm) ratios or FR light intensities in order to investigate growth and flowering responses. Flower initiation and induction were promoted by night break treatment with a low R:FR light source, but was delayed by a high R:FR ratio. The promotion or delay of flower bud formation was accompanied by a decrease or an increase, respectively, in the number of nodes on the main stem at anthesis to the first floret. The difference between date of visible bud with plants under night break treatment and that of the control was approximated with high accuracy by a sigmoid function of the logarithms of R:FR ratio. The threshold R:FR ratio demarcating the promotion and delay of date of visible bud was about 5.3 under the experimental conditions used. The critical R:FR ratios for promotion or delay of visible bud would be about 0.5 and 50.0, respectively. In addition, the time from planting to visible bud was approximated with an exponential function of FR light intensity. The maximum acceleration of date of visible bud by long-day treatment would be 20 days, and the critical FR light intensity would be 2.0 μmol m⁻² s⁻¹. It is concluded that growth and flowering of E. grandiflorum can be regulated by long-day treatment using light sources with different R:FR ratios or FR light intensities.     

C-reactive protein concentration in canine idiopathic polyarthritis

To investigate the clinical utility of C-reactive protein (CRP) in idiopathic polyarthritis (IPA), its concentration was measured in dogs with IPA. The CRP concentration was markedly increased in all the IPA dogs at the time of diagnosis and decreased significantly in response to the initial corticosteroid treatment; this indicated that CRP can be used as an index for therapeutic response in IPA cases. Furthermore, at 6 months after the diagnosis, a significant association was observed between the CRP concentration at follow-up (6-13 days after the treatment was started) and the frequency of medication ("no or seldom-medicated (NSM)" groups or "continuing medication (CM)" groups). These results suggest that the initial response of CRP to corticosteroid treatment may be a prognostic factor of canine IPA.     

A unique isolate of Malassezia from a cat

An isolate of Malassezia from a cat with otitis externa was examined mycologically as well as molecularly. The isolate was similar to M. sympodialis in morphological and biochemical characteristics. In molecular analysis, however, it differed from the 7 species of Malassezia previously reported. Therefore, this clinical isolate from a cat might be a new species of Malassezia.