Development and characterization of 21 polymorphic microsatellite loci in the aphidophagous gall midge, <Emphasis Type="Italic">Aphidoletes aphidimyza</Emphasis> (Diptera: Cecidomyiidae)

We have developed and characterized 21 microsatellite markers in the aphidophagous gall midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). All 21 loci tested were polymorphic: the number of alleles ranged from 2 to 17. Allelic richness and observed heterozygosities were higher in females than in males. Several loci had no heterozygosity in males, suggesting that the loci were located on sex chromosomes or E-chromosomes, common to cecidomyiids. The high polymorphism detected in this study suggests the markers will be of value in analyzing genetic structure of field populations.     

Effects of remnant primary forests on ant and dung beetle species diversity in a secondary forest in Sarawak, Malaysia

Tropical landscape structures have been transformed into mosaic structures consisting of small patches of primary and secondary forests, and areas of other land use. Diversity of insect assemblages is often higher in primary forests than in surrounding secondary forests. However, little is known about how the primary forests affect diversity in surrounding secondary forests in a landscape. In Sarawak, Malaysia, the typical landscape in areas from which lowland tropical rainforests had originally spread consists mainly of primary and secondary forests, with small areas of cultivation. In this study, we examined how the proportion of remnant primary forests in a landscape affects species diversity and species composition of ants and dung beetles in Macaranga-dominated secondary forests. The proportions were quantified based on remote-sensing data at various spatial scales, ranging from 100- to 5,000-m radius from each of the target forests. We found that the proportions of remnant primary forests within a 100-m radius had a significant positive effect on ant species diversity, and those within 100-, 300-, and 500-m radii significantly affected species compositions. However, the proportions of remnant primary forests had no significant relationship with dung beetle diversity, while those within 100- and 1,000-m radii had significant effects on species composition. The different responses to the remnant primary forests are likely to be related to differences in the movement and dispersal traits between the two taxa.     

A Reduction in Age-Enhanced Gluconeogenesis Extends Lifespan

The regulation of energy metabolism, such as calorie restriction (CR), is a major determinant of cellular longevity. Although augmented gluconeogenesis is known to occur in aged yeast cells, the role of enhanced gluconeogenesis in aged cells remains undefined. Here, we show that age-enhanced gluconeogenesis is suppressed by the deletion of the tdh2 gene, which encodes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a protein that is involved in both glycolysis and gluconeogenesis in yeast cells. The deletion of TDH2 restores the chronological lifespan of cells with deletions of both the HST3 and HST4 genes, which encode yeast sirtuins, and represses the activation of gluconeogenesis. Furthermore, the tdh2 gene deletion can extend the replicative lifespan in a CR pathway-dependent manner. These findings demonstrate that the repression of enhanced gluconeogenesis effectively extends the cellular lifespan.     

Construction of Transducing Phage ρ 11 Containing α-Amylase Structural Gene of Bacillus subtilis

We constructed a reporter system to detect a superoxide-generating methyl viologen using SoxRS of Escherichia coli and GFP of Aequorea victoria. E. coli carrying this plasmid exhibited strong fluorescence when grown in the presence of a superoxide-generating reagent methyl viologen. The fluorescence intensity observed in the stationary phase culture of the transformant increased in response to the methyl viologen concentration in a range of 0.01 μM to 10 μM.     

An Assay of Sterigmatocystin and Related Metabolites of Aspergillus versicolor by High Speed Liquid Chromatography

Among several type cultures that assimilated 1-hexadecene, Corynebacterium equi IFO 3730 was found to best accumulate 1, 2-epoxyhexadecane. The purified product exhibited +9.64 (c = 3.71, n-hexane) and was confirmed to have the (R) absolute configuration by correlating to known analogous compounds. The optical purity was determined to be 100% by PMR measurement of 1-methoxy-2-hexadecanol which was derived stereospecifically from the epoxide. The highest yield (41 % based on consumed 1-hexadecene) was achieved when 2.0% of octane and 0.1 % of Tween 80 were added to the medium containing 0.5 % of the olefin. C. equi also assimilated terminal olefins other than 1-hexadecene and produced the corresponding epoxides from substrates which have carbon chains longer than fourteen.     

Identification of GA3 in Neurospora crassa and Its Changes during Conidial Germination and Mycelial Growth

GA₃ was identified as a major GA in Neurospora crassa by gas chromatography/selected ion monitoring (GC/SIM) and its content was measured at various stages (0~96 hr after inoculation) of conidial germination and mycelial growth. The GA₃ content in the fungus was 190ng/g dry weight at the initial stage and then decreased rapidly; that per liter culture decreased soon after the inoculation, and then increased to 17.6 ng 96 hr after inoculation. The GA₃ concentration in the culture medium was around 10^?11 m throughout the 96-hr incubation. The physiological role of endogenous GA in Neurospora crassa is also discussed.     

Isolation and Characterization of Two Plasmids in Bacillus subtilis var. amylosacchariticus

Five bufadienolides (1-5) isolated from the leaves of Kalanchoe pinnata and K. daigremontiana×tubiflora (Crassulaceae) were examined for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation in Raji cells induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. All bufadienolides showed inhibitory activity, and bryophyllin A (1) exhibited the most marked inhibition (IC₅₀=0.4 μM) among the tested compounds. Bryophyllin C (2), a reduction analogue of 1, and bersaldegenin-3-acetate (3) lacking the orthoacetate moiety were less active. These results strongly suggest that bufadienolides are potential cancer chemopreventive agents.     

Calcitonin gene-related peptide regulates mitogen-activated protein kinase pathway to decrease transforming growth factor β1-induced hepatic plasminogen activator inhibitor-1 mRNA expression in HepG2 cells

Transforming growth factor (TGF) β1-induced plasminogen activator inhibitor (PAI)-1 is one of factors associated with the development of hepatic fibrosis. Calcitonin gene-related peptide (CGRP) shows hepatoprotective effect during hepatic injuries, including fibrosis. However, the effects of CGRP on PAI-1 expression induced by TGFβ1 are unknown. In this study, we investigated the effect of CGRP on TGFβ1-induced PAI-1 expression and its regulatory mechanisms in HepG2 cells. CGRP inhibited TGFβ1-induced PAI-1 expression. H89, a protein kinase A inhibitor, abolished the inhibition of TGFβ1-induced PAI-1 expression by CGRP. TGFβ1 activated mitogen-activated protein kinase (MAPK), including extracellular signal-regulated kinase, c-jun NH₂-terminal kinase, and p38, and this activation was abolished by CGRP. These results show that the CGRP-induced cAMP/PKA activation suppresses activation of MAPK induced by TGFβ1, leading to decreased PAI-1 expression in HepG2 cells.     

Bio-functional pickles that reduce blood pressure of rats

Addition of γ-aminobutyric acid (GABA) and angiotensin converting enzyme (ACE)-inhibitory peptides to the pickles was studied in order to develop a new type of pickles that reduce blood pressure. Based on the outcome of these studies, a new type of fermentation bed composed of rice bran and white miso has been successfully developed. The advantage of such pickles is that they not only contain both GABA and ACE-inhibitory peptides, but also that their taste and flavor are excellent, with colors close to the original ones. The new type of pickles could temporarily reduce blood pressure in two types of rats, spontaneously hypertensive rats and NaCl-sensitive model rats. Thus, the newly developed pickles appear to be beneficial for pickle business.