Determination of nitrobenzene oxidation products by GC and1H-NMR spectroscopy using 5-iodovanillin as a new internal standard

The nitrobenzene oxidation method was modified to obtain more reproducible data and more structural information about lignin, not only by gas chromatography (GC) but also by proton nuclear magnetic resonance (¹H-NMR) spectroscopy for quantitative determination of the oxidation products and to simplify the procedures. The nitrobenzene oxidation mixture was directly extracted after acidification without preextraction of by-products. The direct extraction made the extractive step easy and gave reproducible data. 5-Iodovanillin was selected as a new internal standard. The reason for this selection was that 5-iodovanillin did not exist in the nitrobenzene oxidation products from any plant species and had an aldehyde group whose peak did not overlap with the other aldehyde peaks on an¹H-NMR spectrum. Thus, the use of 5-iodovanillin enabled us to quantifyp-hydroxybenzaldehyde, vanillin, and syringaldehyde in oxidation products on the basis of¹H-NMR analysis as well as GC. Furthermore, more information about the condensed structure of lignin was derived by comparing the¹H-NMR and GC analyses.Part of this work was presented at the 42nd Annual Meeting of the Lignin Symposium, Sapporo, October 1997     

Seasonal patterns of downstream movement in partially migratory stream‐dwelling Dolly Varden

Migration is a common behaviour among salmonids, but not all individuals within a population migrate, instead becoming residents and remaining in their natal streams. This phenomenon, known as “partial migration,” is well studied among sea‐run and lake‐run populations; however, the lower migratory benefits and costs for mainstem‐run individuals question whether the same mechanisms can be applied for stream‐dwelling salmonids. In this study, we investigated main stem‐run timing and body condition of partially migratory stream‐dwelling Dolly Varden charr (Salvelinus malma) in the Shiisorapuchi River in central Hokkaido, Japan. Based on commonly observed patterns in sea‐run and lake‐run salmonids, we predicted that migration would occur during spring and consist predominantly of age 1 +  females with some small subordinate males. Traps were placed at the mouths of two small tributaries along the Shiisorapuchi River periodically from May to December 2015. Adipose fin samples were taken for DNA to sex the individuals. Consistent to initial predictions, downstream movement occurs only once in the spring, dominated by age‐1 +  females. Trapped fishes (presumed migrants) were slightly longer but slimmer compared to the fish caught in the tributaries (presumed resident), which may be equivalent to smolts in anadromous populations. Regardless of migratory habitat, mechanisms driving partial migration in salmonids may be the same as long as production between natal stream and feeding habitats is significantly different.     

Solid-state selective (13)C excitation and spin diffusion NMR to resolve spatial dimensions in plant cell walls

The average spatial dimensions between major biopolymers within the plant cell wall can be resolved using a solid-state NMR technique referred to as a (13)C cross-polarization (CP) SELDOM (selectively by destruction of magnetization) with a mixing time delay for spin diffusion. Selective excitation of specific aromatic lignin carbons indicates that lignin is in close proximity to hemicellulose followed by amorphous and finally crystalline cellulose. (13)C spin diffusion time constants (T(SD)) were extracted using a two-site spin diffusion theory developed for (13)C nuclei under magic angle spinning (MAS) conditions. These time constants were then used to calculate an average lower-limit spin diffusion length between chemical groups within the plant cell wall. The results on untreated (13)C enriched corn stover stem reveal that the lignin carbons are, on average, located at distances ～0.7-2.0 nm from the carbons in hemicellulose and cellulose, whereas the pretreated material had larger separations.     

Chemical structural elucidation of total lignins in woods I: fractionation of the lignin in residual wood meal after extraction of milled wood lignin

Residual wood meal after extraction of milled wood lignin (WMEM) ofEucalyptus globulus was extracted with alkali and LiCl/N,N-Dimethylacetamide (DMAc). These agents dissolve mainly hemicellulose and cellulose, respectively. The extractability of WMEM in alkali solutions was influenced by the degree of swelling of the cellulose. Under good swelling conditions, considerable amounts of cellulose and lignin were extracted with the hemicellulose. Maximum extractability was about 60% of the WMEM under optimum conditions (3 M or 5 M LiOH or 3M NaOH solution). Some portion of cellulose was extracted with LiCl/DMAc at room temperature. Thus, lignin inE. globulus WMEM was divided into three fractions: hemicellulose-lignin fraction, cellulose-lignin fraction, and insoluble-lignin fraction.Part of this work was presented at the 49th annual meeting of the Japan Wood Research Society, Tokyo, April 1999; and at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000     

Insect prothoracicotropic hormone: evidence for two molecular forms

In an insect, the tobacco hornworm Manduca sexta, the cerebral neuropeptide prothoracicotropic hormone (PTTH), the primary effector of postembryonic development, exists as two molecular forms. These two PTTH's elicit characteristic in vitro dose responses of activation of prothoracic glands from different developmental stages, an indication that during development the glands change in their sensitivity to the neurohormones. Both PTTH's are active in a specific in situ bioassay. Since they may be released in situ at stage-specific times to evoke distinctly different developmental responses, the PTTH neuroendocrine axis appears to be an effective system for determining the functions of molecular forms of a neurohormone in the regulation of growth and development.     

Production of a new low-caffeine hybrid coffee and the biochemical mechanism of low caffeine accumulation

The GCAs are new tetraploid interspecific hybrids developed in Madagascar from Coffea eugenioides, C. canephora and C. arabica. Selected GCA having genotype UF1023 contained 0.37% DW caffeine and no detectable theobromine in green beans. Low caffeine accumulation in GCA plants is due mainly to the low biosynthetic activity of purine alkaloids, possibly the extremely weak N-methyltransferase reactions in caffeine biosynthesis. No significant catabolic activity of caffeine was found in GCA-UF1023, in common with almost all coffee plants including C. arabica.     

Statistical Analysis of Metal Concentrations in a Sediment Core to Reveal Influences of Human Activities on Atmospheric Environment for 200 Years

Metal concentrations in a sediment core from a moat outside Osaka Castle in Japan were measured by ICP-MS following a microwave extraction method. Concentrations of metals in the sediment core samples peaked around 1945 when the World War II ended. This is in part due to great air raids on Osaka. In cluster analysis using metal concentrations, the fractions of sediment core samples were classified into two groups, representing natural sources (1795–1915) and anthropogenic sources (1922–1976), respectively. Results of lead isotope ratios also showed the anthropogenic influences from 1915 by changing the ratio values compared to those of natural sources. In addition, several components contributing to metal concentrations were identified by principal component analysis. The main component was controlled by natural sources and a decrease of the component score corresponded to an increased influence from anthropogenic sources. The largest impact of anthropogenic sources was shown around 1945.     

Removal of sperm tail using trypsin and pre-activation of oocyte facilitates intracytoplasmic sperm injection in mice and rats

We examined various methods to enhance the accessibility of intracytoplasmic sperm injection (ICSI) technology to more users by making the technique easier, more efficient, and practical. First, the methods for artificially removing the mouse sperm tail were evaluated. Trypsin treatment was found to efficiently remove the sperm tails. The resultant sperm cells had a lower oocyte activation capacity; however, the use of activated oocytes resulted in the same fecundity as that of fresh, untreated sperm. Pre-activated oocytes were more resistant to physical damage, showed higher survival rates, and required less time per injection. Testing this method in rats yielded similar results, although the oocyte activation method was different. Remarkably, this method resulted in higher birth rates of rat progeny than with conventional methods of rat ICSI. Our method thereby streamlines mouse and rat ICSI, making it more accessible to laboratories across many disciplines.