Characterization and expression of non‐polymorphic liver expressed antimicrobial peptide 2: LEAP‐2 in the Japanese quail,Coturnix japonica

Liver‐expressed antimicrobial peptide 2 (LEAP‐2) is a cationic peptide that plays an important role in innate immunity for host defense. The aim of this study was to characterize the LEAP‐2 gene in the Japanese quail (Coturnix japonica). Japanese quail LEAP‐2 (CjLEAP‐2) was identified from the Japanese quail draft genome database by a local BLAST analysis using chicken LEAP‐2 (GgLEAP‐2). The exon‐intron structure of CjLEAP‐2, analyzed from three quails, is composed of three exons, as is the chicken LEAP‐2 homolog (GgLEAP‐2). An analysis of the coding sequence revealed that CjLEAP‐2 is 231 bp long, like GgLEAP‐2, and 93% identical to GgLEAP‐2 at the nucleic acid level. The predicted amino acid sequence of CjLEAP‐2 contained the liver‐expressed antimicrobial peptide 2‐precursor domain and four cysteine residues characteristic of the LEAP‐2 protein. The amino acid sequence of the mature peptide of CjLEAP‐2 was 100% identical to that of GgLEAP‐2. We confirmed that CjLEAP‐2 was transcribed in at least seven tissues, including the digestive system. Additionally, the mature peptide region of CjLEAP‐2 exhibited no polymorphisms in 99 quails from six strains. Taken together, these findings indicate that CjLEAP‐2 is non‐polymorphic and therefore, it likely plays an important role in the innate immunity of quail as it does in chicken.     

Enhancement of saccharification by overexpression of poplar cellulase in sengon

Lignocellulosic material from trees has great potential to form the basis of the second generation for bioethanol production because trees produce most of the biomass on the earth. We modified the wall structure of sengon (Paraserianthes falcataria) through overexpression of poplar cellulase in the cell walls. The overexpression did not decrease cellulose content but caused a decrease in xyloglucan bound to the walls. The level of saccharification and successive ethanol production was increased in the wood of the transgenic sengon overexpressing poplar cellulase compared with that of the wild type plant, and even after delignification of the wood. We propose that a xyloglucan intercalated into cellulose microfibrils could be one of the recalcitrant components in the saccharification of lignocelluloses.     

Proteomic analysis of the G-layer in poplar tension wood

Angiosperm trees bend their stems by forming tension wood at the upper side of leaning stems. Most tension wood has a cellulose-rich G-layer in the innermost surface of the fiber cell wall. Strong tensile stress is considered to occur in the G-layer. This study undertook to identify the proteins involved in G-layer formation and function through a proteomic analysis of G-layer-localized protein. G-layers of poplar were loosened by sonication and isolated as doughnut-shaped pieces of thinly sliced transverse sections. The proteins, once extracted with urea/detergent solution, were separated by two-dimensional polyacrylamide gel electrophoresis, and 110 spots were subjected to liquid chromatography tandem mass spectrometry (LC/MS/MS). A database search for these spots’ mass spectrum patterns identified 72 proteins. In addition, all peptide digestion mixtures of G-layer proteins were separated by strong cation exchange chromatography and 39 proteins were identified using LC/MS/MS analysis. Proteins involved in wall formation, such as lignin biosynthesis-related protein, xyloglucan endotransglucosylase, and fasciclin-like arabinogalactan protein, were notably detected in the G-layer.     

Effect of pressure on organic acid production from Japanese beech treated in supercritical water

The effect of pressure as described by density was studied on organic acid production from Japanese beech (Fagus crenata) treated in supercritical water. At a reaction temperature of 380°C, the maximum yield of organic acids was 35% at a pressure of 30 MPa (density of water: 0.53 g/ml) for 1 min in a batch-type system. Furthermore, the yield of organic acids decreased with increasing reaction pressure. It was also found that fragmented products from sugars such as methylglyoxal and glycolaldehyde could be more easily converted to organic acids than dehydrated products such as furfural and 5-hydroxymethyl furfural. This result suggests that organic acids can be mainly derived from fragmented products. Part of this article was presented at the 55th Annual Meeting of the Japan Wood Research Society (2005), and at the 17th Annual Meeting of Japan Institute of Energy (2007)     

Genetic identification and morphology of naturally spawned eggs of the Japanese eel Anguilla japonica collected in the western North Pacific

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.     

Occurrence of heavy metals in the sediments of Uranouchi Inlet,Kochi prefecture,Japan

The present status of contamination by heavy metals, and the impact of cage culture on sediments at the Uranouchi Inlet, Kochi Prefecture, Japan, were investigated from two stations influenced by intensive aquaculture and a control station, during May–July 2006. The moisture content of the sediments at the aquaculture stations was over 65%, and the organic matter was always over 100 mg/g dry wt. In contrast, the highest moisture content and organic matter at the control station was 45.5% and 62.9 mg/g dry wt, respectively. Concentrations of zinc (Zn) (178 ± 4.8 mg/kg dry wt) and copper (Cu) (125 ± 1.2 mg/kg dry wt) were highest at the aquaculture stations. Lead (Pb) was highest (50.7 ± 0.77 mg/kg dry wt) at the aquaculture station though it was as high as 33.2 ± 0.77 mg/kg dry wt at the control station. One-way ANOVA showed that the differences in concentrations of Zn and Cu in sediments from the aquaculture and control stations were highly significant (P = 0.01), whereas Pb showed no such trend. Occurrence of a large fraction of labile Zn (56.1%) and Cu (40.3%) in these sediments warrants attention. Although factors other than metals may explain the distribution observed, the information presented here may be useful in predicting long-term effects of heavy metal contamination from aquaculture in the marine environment.     

Cellulose esters as compatibilizers in wood/poly(lactic acid) composite

One of the important issues relevant to wood/plastic composite molding is the compatibility between hydrophilic wood and hydrophobic plastic. Polyolefins modified with maleic anhydride, which have been frequently used for wood and polyolefin composites, are not effective for poly(lactic acid) composites. Because compounds with both hydrophilic and hydrophobic groups are potential compatibilizers, cellulose esters of several carboxylic acids, RCOO-cellulose (R = CH₃, C₂H₅, C₃H₇, C₄H₉, C₅H₁₁, C₁₁H₂₃), were synthesized and their effects as compatibilizers of wood/poly(lactic acid) composites (WPLC) were examined in this study. The mechanical properties of WPLC were improved with a small amount of added cellulose esters, especially cellulose butyrate or cellulose valerate. The relevant effect of added cellulose esters on the thermal properties of wood/PLA composite was analyzed by measuring dynamic viscoelasticity. Part of this report was presented at the 56th Annual Meeting of Japan Wood Research Society, Akita, August 2006 and the 9th International Conference on Frontiers of Polymers and Advanced Materials, Cracow, Poland, July, 2007     

Effects of density profile of MDF on stiffness and strength of nailed joints

Nail-head pull-through, lateral nail resistance, and single shear nailed joint tests were conducted on medium density fiberboard (MDF) with different density profiles, and the relations between the results of these tests and the density profiles of MDF were investigated. The maximum load of nail-head pull-through and the maximum load of nailed joints were little affected by the density profile. However, the ultimate strength of lateral nail resistance, the stiffness, and the yield strength of nailed joints were affected by the density profile of MDF and showed high values when the surface layer of the MDF had high density. It is known that bending performance is also influenced by density profile. Therefore, the stiffness and the yield strength of nailed joints were compared with the bending performance of MDF. The stiffness of nailed joints was positively correlated with the modulus of elasticity (MOE); in the case of CN65 nails, the initial stiffness of joints changed little in response to changes in MOE. The yield strength of nailed joints had a high positive correlation with the modulus of rupture (MOR). The stiffness and the yield strength of nailed joints showed linear relationships with MOE and MOR, respectively.     

Analysis of urbanization characteristics causing farmland loss in a rapid growth area using GIS and RS

This study aims to analyze the spatial characteristics of urbanization in a rapid growth area. Remote sensing (RS) and geographic information system (GIS) techniques were used to classify land uses in 1986 and 1996, the most rapid growth period. The two land-use maps were classified with accuracies of 90% and 86% in verification areas for 1986 and 1996, respectively, by a supervised maximum likelihood method. Urbanization in the greenbelt within the study area was restricted, while outside the greenbelt it occurred rapidly. Fifty seven percent of the total increase in built-up area in the study area was concentrated in a 10-km GIS-defined buffer zone adjacent to the greenbelt. The priority for new residential development was put on areas with slopes of less than 15 degrees and potential farmland in the areas surrounding Seoul. In the priority area, farmland loss was very significant. These results show that although the environmental protection of ecologically valuable land was well enforced by the greenbelt from urbanization, farmland was not conserved in comparison to new residential development in the rapid growth area.