Non‐targeted analyses of animal plasma: betaine and choline represent the nutritional and metabolic status

Simple liquid chromatography–mass spectrometry (LC‐MS) was applied to non‐targeted metabolic analyses to discover new metabolic markers in animal plasma. Principle component analysis (PCA) and partial least squares–discriminate analysis (PLS‐DA) were used to analyse LC‐MS multivariate data. PCA clearly generated two separate clusters for artificially induced diabetic mice and healthy control mice. PLS‐DA of time‐course changes in plasma metabolites of chicks after feeding generated three clusters (pre‐ and immediately after feeding, 0.5–3 h after feeding and 4 h after feeding). Two separate clusters were also generated for plasma metabolites of pregnant Angus heifers with differing live‐weight change profiles (gaining or losing). The accompanying PLS‐DA loading plot detailed the metabolites that contribute the most to the cluster separation. In each case, the same highly hydrophilic metabolite was strongly correlated to the group separation. The metabolite was identified as betaine by LC‐MS/MS. This result indicates that betaine and its metabolic precursor, choline, may be useful biomarkers to evaluate the nutritional and metabolic status of animals.     

Characterization of neuron-like cells derived from canine bone marrow stromal cells

Regenerative therapy using bone marrow stromal cells (BMSCs) has begun to be clinically applied in humans and dogs for neurological disorders such as spinal cord injury. Under appropriate conditions in vitro, BMSCs differentiate into neuronal cells, which may improve the effects of regenerative therapy. In this study, we evaluated canine neuron-like cells (NLCs) derived from BMSCs. We speculated on their suitability for neuro-transplantation from the point of view of their morphological features, long-term viability, abundant availability, and ability to be subcultured. Canine NLCs were differentiated as follows: third-passage BMSCs were maintained in pre-induction medium containing 2-mercaptoethanol and dimethylsulfoxide for 5 h, and then cells were transferred to neuronal induction medium containing fetal bovine serum, basic fibroblast growth factor, epidermal growth factor, dibutyryl cyclic AMP, and isobutylmethylxanthine for 7 or 14 days. Canine NLCs fulfilled the transplantation criteria and expressed markers of both immature neurons (nestin, 84.7 %) and mature neuronal cells (microtubule-associated protein-2, 95.7 %; βIII-tubulin protein, 12.9 %; glial fibrillary acidic protein, 9.2 %). These results suggest that canine BMSCs can be induced to differentiate into neuronal cells and may be suitable for neuro-transplantation. This study may provide information for improving cellular therapy for neurological diseases.     

In-crisis delivery rate: a novel measure of success in communal water management

Of the plethora of studies that discuss requirements for successful irrigation management, few pay close attention to what actually happens when the supply of water becomes extremely low. Such an oversight in the literature is unfortunate, because this is precisely when management matters. To understand what separates success from failure in irrigation management at times of critical water shortages, the authors conducted emergency fieldwork in February 2010 along four major irrigation channels in a drought-stricken rice-growing area within the Chinese province of Yunnan. Separately, satellite images of the four villages dated February 2009 and February 2010 were jointly analysed to produce a novel indicator for a village’s success in water management. Called the in-crisis delivery rate, this indicator compares water delivery between normal and drought years and directly evaluates performance in water management during crisis periods. The results led to an unexpected discovery that, contrary to common expectations, the only village of the present study that deployed a traditional earthen water channel secured substantially more water throughout the drought period than those with concrete-lined channels. It is hypothesised that the labour intensive, rather than capital intensive, nature of repair work of the earthen channel enabled flexible operations, and hence had the comparative advantage under a skilled management team. This result confirms the importance of daily maintenance work, which tends to occur less often after modernisation of water paths.     

Neutral and acidic milk oligosaccharides of the striped skunk (Mephitidae: Mephitis mephitis)

The biological significance of the tremendous variation in proportions of oligosaccharides and lactose among mammalian milks is poorly understood. We investigated milk oligosaccharides of the striped skunk (Mephitis mephitis) and compared these results to other species of the clade Mustelida. Individual oligosaccharides were identified by proton nuclear magnetic resonance spectroscopy. In the striped skunk, six oligosaccharides were identified: isoglobotriose, 2′‐fucosyllactose, A‐tetrasaccharide, Galili pentasaccharide, 3′‐sialyllactose and monosialyl monogalactosyl lacto‐N‐neohexaose. Four of these have been found in related Mustelida and the other two in more distantly related carnivorans. The neutral and acidic oligosaccharides derive from three core structures: lactose (Gal(β1–4)Glc), lacto‐N‐neotetraose (Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc) and lacto‐N‐neohexaose (Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc).     

L‐carnitine improves hydrogen peroxide‐induced impairment of nuclear maturation in porcine oocytes

We investigated the effect of oxidative stress induced by hydrogen peroxide (H₂O₂) on lipid peroxide (LPO) level and nuclear maturation in porcine oocytes cultured with or without cumulus cells. After 22 h of pre‐culture, oocytes with attached cumulus cells (COC group) or denuded oocytes (DO group) were cultured with H₂O₂, and intra‐oocyte H₂O₂ and LPO levels were quantitatively analyzed using immunofluorescence. This is the first report evaluating LPO levels in porcine oocytes. After H₂O₂ supplementation, the DO group showed severe accumulation of H₂O₂ and LPO in the oocytes. Similarly, while inhibition of progression of nuclear maturation was observed in both groups, the effect was more severe in the DO group. These results demonstrate that cumulus cells reduce the accumulation of H₂O₂ stress in oocytes. Furthermore, we attempted to reduce the oxidative stress by H₂O₂ with L‐carnitine, a H₂O₂ scavenger. L‐carnitine decreased H₂O₂ and LPO levels in the oocytes in both groups, and improvement in the progression of impaired nuclear maturation was observed. These effects were different by the presence of cumulus cells. Our results provide that L‐carnitine is useful for alleviating H₂O₂‐induced oxidative stress by reducing LPO levels and improving the progression of nuclear maturation.     

Diversity of S-RNase genes and S-haplotypes in Japanese plum (Prunus salicina Lindl.)

Summary

This report demonstrates the diversity of S-haplotypes in Japanese plum by molecular cloning of genomic DNAs and cDNAs that encode S-RNases. Nine different DNA fragments, designated as S^a–Sⁱ, were obtained from 17 Japanese plum cultivars by PCR with an S-RNase gene-specific primer set, Pru-C2 and PCE-R. Eleven different S-haplotypes were found in these cultivars. The banding patterns obtained with another S-RNase gene-specific primer set, Pru-T2 and PCE-R, corresponded to the S-haplotypes predicted from the Pru-C2 and PCE-R primer set. Several cultivars had the same S-haplotypes. Partial genomic DNAs for eight S-RNase genes and cDNAs for two S-RNases were cloned and sequenced. Deduced amino acid sequences contained conserved regions among the rosaceous S-RNases. Comparisons of the sequences from cDNAs and genomic DNAs revealed the presence of two introns in the S-RNase genes of Japanese plum as in other Prunus S-RNase genes. Pollination incompatibility groups and self-compatibility in Japanese plum were discussed with reference to the S-haplotypes.     

The suitable harvesting season and the part of moso bamboo (Phyllostachys pubescens) for producing binderless boards

The properties of the binderless boards of moso bamboo depending on the harvest seasons and the parts of the height were reported and the optimum harvesting conditions investigated. The binderless boards were prepared from the powdered bamboo harvested each month from June to May, and the parts in height. The hot water extract (HWE), lignin, α-cellulose, and hemicelluloses contents were examined. The board properties were evaluated with internal bonding (IB), water absorption (WA), and thickness swelling (TS). From the experiment, the boards prepared between March and October had higher HWE content and higher IB and lower WA and TS than the ones prepared in other months. The board from the top part showed lower IB and higher WA and TS than the bottom and the middle. The boards prepared from the residue after extraction of HWE showed lower IB than the unextracted samples. These results indicated that for producing board from bamboo, the suitable harvesting season is when HWE contents are higher and that the suitable parts in height for harvesting are the bottom and the middle where lignin contents are not low.     

Diversification and genetic differentiation of cultivated melon inferred from sequence polymorphism in the chloroplast genome

Molecular analysis encouraged discovery of genetic diversity and relationships of cultivated melon (Cucumis melo L.). We sequenced nine inter- and intra-genic regions of the chloroplast genome, about 5500 bp, using 60 melon accessions and six reference accessions of wild species of Cucumis to show intra-specific variation of the chloroplast genome. Sequence polymorphisms were detected among melon accessions and other Cucumis species, indicating intra-specific diversification of the chloroplast genome. Melon accessions were classified into three subclusters by cytoplasm type and then into 12 subgroups. Geographical origin and seed size also differed between the three subclusters. Subcluster Ia contained small-seed melon from Southern Africa and South and East Asia and subcluster Ib mainly consisted of large-seed melon from northern Africa, Europe and USA. Melon accessions of subcluster Ic were only found in West, Central and Southern Africa. Our results indicated that European melon groups and Asian melon groups diversified independently and shared the same maternal lineage with northern African large-seed melon and Southern African small-seed melon, respectively. Cultivated melon of subcluster Ic may have been domesticated independently in Africa. The presence of 11 cytoplasm types in Africa strongly supported African origin of cultivated melon and indicated the importance of germplasm from Africa.     

Characterization of bagasse binderless particleboard manufactured in high-temperature range

This paper describes the features of binderless particleboard manufactured from sugarcane bagasse, under a high pressing temperature of 200–280 °C. Mechanical properties [i.e., modulus of rupture (MOR) and elasticity (MOE) in dry and wet conditions, internal bonding strength (IB)] and dimensional stability [i.e., thickness swelling (TS)] of the board were evaluated to investigate the effect of high pressing temperature. Recycled chip binderless particleboards were manufactured under the same conditions for comparison, and particleboards bonded with polymeric methylene diphenyl diisocyanate (PMDI) resin were manufactured as reference material. The target density was 0.8 g/cm³ for all of the boards. The results showed that the mechanical properties and dimensional stability of both types of binderless boards were improved by increasing the pressing temperature. Bagasse showed better performance than that of recycled chip as a raw material in all evaluations. Bagasse binderless particleboard manufactured at 260 °C had an MOE value of 3.5 GPa, which was equivalent to the PMDI particleboard, and a lower TS value of 3.7 % than that of PMDI particleboard. The MOR retention ratio under the dry and wet conditions was 87.0 %, while the ratio for the PMDI particleboard was only 54.6 %. The obtained results showed the possibility of manufacturing high-durability binderless particleboard, with good dimensional stability and water resistance, which previously were points of weakness for binderless boards. Manufacturing binderless boards under high temperature was effective even when using particles with poor contact area, and it was possible to express acceptable properties to allow the manufacture of particleboards. Further chemical analysis indicated a contribution of a saccharide in the bagasse to the improvement of the board properties.