Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)

We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.     

Optimum planting density for Japanese oak (Quercus mongolica var.grosseserrata) based on spacing experiment with systematic design

A spacing experiment for Japanese oak (Quercus mongolica var.grosseserrata) with Nelder's design have been studied at the Kyushu University Forest in Hokkaido, since 1975. The oak seedlings were planted according to systematic spacings which were 20 levels varying gradually from 40,816 to 1,086 seedlings per hectare with concentric circular arcs (22 in number) and spokes (41 in number). In 1992 when the planted oak seedlings were 18-year-old, DBH, crown diameter, total height and clear length of the survival trees were measured. Based on the results obtained from analyzing the relationships between planting density and the mean values of each block, the optimum planting density should be 7,241 seedlings per hectare. The title is tentative translation from the original Japanese title by the authors of this paper.     

Species composition of Modellidae and Cerambycidae (Coleoptera) in a coppice woodland

A continuous sampling of canopy beetles was carried out to determine variations in the abundance, species diversity, richness, and composition of the Mordellidae and Cerambycidae in a coppice woodland. Changes in the abundance and the species richness were monitored at three heights in the forest throughout the season in 1999, using yellow and blue water pan traps. The results showed significant variations in the abundance of Mordellidae among the canopy layers, while little variation was found for Cerambycidae. The abundance, species diversity, and richness were generally greater in summer. The results showed distinct species compositions in both families among layers.     

GC-MS and IR spectroscopic analyses of the lignin-derived products from softwood and hardwood treated in supercritical water

Softwood (Cryptomeria japonica) and hardwood (Fagus crenata) were treated in supercritical water (380°C, 100 MPa) for 8 s. The treated woods were fractionated to the water-soluble portion, methanol-soluble portion, and methanol-insoluble residues. For the methanol-soluble portion, which mainly consisted of lignin-derived products, gel permeation chromatography (GPC) and gas chromatographic-mass spectrometric (GC-MS) analyses were conducted to clarify the molecular weight distribution and to identify the monomeric products, respectively. GPC analysis revealed that the methanol-soluble portion contains monomeric and some oligomeric products. GC-MS analysis identified 19 guaiacyl compounds in the methanol-soluble portion from softwood, and 15 syringyl monomeric compounds in the methanol-soluble portion from hardwood. The structures of identified products included not only phenyl propane (C₆—C₃) units but also C₆—C₂ and C₆—C₁ units. In addition, the infrared spectra suggested that the methanol-soluble portion maintains the typical structure of lignin, although it is rich in condensed-type linkages with some changes in the propyl side chain. These results indicate that the supercritical water treatment cleaves not only ether linkages but also part of the propyl chains in lignin to give various aromatic compounds.     

Gas chromatographic and mass spectrometric (GC-MS) analysis of lignin-derived products from <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> treated in supercritical water

Sugi (Cryptomeria japonica D. Don) wood was treated with supercritical water (374°C, 22.1MPa), and fractionated into the water-soluble portion, the methanol-soluble portion, and the methanol-insoluble residue. The methanol-soluble portion mainly consisted of the lignin-derived products. To characterize the compounds in the methanol-soluble portion, gel permeation chromatographic (GPC) and gas chromatographic-mass spectrometric (GC-MS) analyses were performed. The GPC analysis indicated that the methanol-soluble portion contained lignin-derived monomeric and dimeric products. GC-MS analysis detected 31 products which were expected to be monomeric compounds, and 18 of these were identified to be guaiacol, methylguaiacol, ethylguaiacol, vinylguaiacol, eugenol, propylguaiacol, vanillin, cis-isoeugenol, homovanillin, trans-isoeugenol, acetoguaiacone, propioguaiacone, guaiacylacetone, 2-methoxy-4-(1-hydroxypropyl)phenol, homovanillic acid, 2-methoxy-4-(prop-1-en-3-one)phenol, coniferyl aldehyde, and ferulic acid. In addition, 22 dimeric products were detected, and 4 of these were believed to be compounds with biphenyl type (5-5), diphenylethane type (-1), stilbene type (-1), and phenylcoumaran type (-5) structures. These results clearly indicated that the methanol-soluble portion included various monomeric and dimeric compounds produced as a result of the cleavage of ether linkages and propyl chains of lignin.     

Development of height curves for Japanese larch in relation to GIS-derived site index

The objectives of this study were: (1) to relate the height growth pattern to the GIS-derived site index, (2) to construct various types of polymorphic height curves, and (3) to select the most accurate height curve for Japanese larch (Larix kaempferi). A total of 14 dominant trees were felled for stem analysis. Richards' function was first fitted to the height/age dataset recorded from stem analyses of 14 trees, and its parameters (i.e., A, k, and m) were then estimated for each sample tree. The relationship between parameter A and the GIS-derived site index was significant, while that of parameters k and m were not significant. The 12 height curve equation forms developed from the Richards' function were then fitted to the dataset consisting of 563 observations of height, age, and GIS-derived site index derived from 14 trees. All height curves were unbiased and provided an equally good fit. The shape of the height curves differed among sites of different site indices. These results indicated that Richards' function was sufficiently flexible to represent the polymorphic height growth pattern of Japanese larch. Finally, the best-fit height curve was selected so that accurate height growth predictions would be possible at any given age and site.