Increased proteolytic susceptibility of carboxypeptidase Y caused by modification of the disulfide zipper

To investigate the structural importance of a "disulfide zipper" motif of carboxypeptidase Y, disulfide-deficient mutant enzymes were expressed in two strains of Saccharomyces cerevisiae. The mutant enzymes were rapidly degraded into fragments by intracellular proteases. Thus, it is concluded that the disulfide zipper is essential in maintaining the structural integrity of CPase Y against proteolytic susceptibility.     

Demonstration of the role of the DnaK chaperone system in assembly of 30S ribosomal subunits using a purified in vitro system

Recently, there has been controversy regarding the ability of the DnaK chaperone system to facilitate Escherichia coli 30S subunit assembly at otherwise nonpermissive conditions. Here, we present additional data indicating that purified DnaK chaperone assembled 30S subunits are functional. Additionally, explanations for reported differences are discussed.     

Acidification of freshwaters by red soil in a subtropical silicate rock area,Okinawa, Japan

Two samples of red soil, one from Gushikawa Recreation Center (GRC) and one from Okinawa Royal Golf Club (ORGC), were examined for particle size distribution, textures, minerals, and chemical compositions. The effects of particle size and grinding of clay minerals on pH, electrical conductivity (EC), and dissolved chemical species were studied in deionized water and river water. The results of red soil solutions were compared with those of acidic waters found in red soil dominated areas. The minimum pH values of soil solutions extracted by deionized water were 4.38–5.36 and 5.16–5.89 and the maximum values of EC were 4.91–16.98mSm^–1 and 3.54–11.23mSm^–1 for GRC and ORGC, respectively. In the river water samples equilibrated with red soils, the minimum pH values were 4.48–5.10 and 4.77–5.91 and the maximum EC values were 19.6–34.2mSm^–1 and 17.5–25.0mSm^–1 for GRC and ORGC, respectively. The values of pH and EC varied with the soil–solution ratio and the particle size. The chemical composition of river water without mixing with red soil shows Na⁺K⁺ and Ca²⁺Mg²⁺. After mixing with red soil, the trend of the concentrations changed to Na⁺K⁺ and Mg²⁺Ca²⁺, which is the same as that of soil solutions in deionized water as well as that of acidic waters found in the red soil area. The pH of the acidic waters was 4.95–5.81 and EC was 7.76–30.0mSm^–1. Laboratory experimental results agreed well with those found in the field in terms of trend of concentrations of the chemical species and pH. Therefore, the results of this study suggest that the low pH and trend of the concentrations of chemical species of the acidic waters found in the red soil dominated areas were the result of the interaction of natural water and red soil.     

Evaluation of a mass screening program for lysinuric protein intolerance in the northern part of Japan

Lysinuric protein intolerance (LPI:MIM 222700) is an autosomal recessive disease characterized by defective transport of the dibasic amino acids. We recently reported a local cluster of LPI in the northern part of Japan (Koizumi et al., 2000). Mutational analysis of the LPI patients in this local cluster revealed they were exclusively homozygous for the R410X mutation. The effectiveness of early intervention with citrulline therapy (200 mg/kg per day) and protein restriction (1.5 g/kg per day) was confirmed in these patients. Mass screening was conducted in 4,568 newborn babies between 1999 and 2002, which was estimated to cover 100% of almost all newborns delivered in the screened area. Forty heterozygous newborns were found (0.88%), leading to an estimated incidence of LPI of 1:51,984. The number of people that required screening to detect one case was 51,984, and the cost for mass screening was 30 cents/person (a total of dollars 15,600). This is comparable to, or even less than, the cost of currently screened diseases in Japan. Therefore, we conclude that a mass screening program for LPI can be introduced effectively and economically into an area where an LPI cluster is located as the result of a founder mutation.     

Diversity and rarity hotspots and conservation of butterfly communities in and around the Aokigahara woodland of Mount Fuji,central Japan

In central Japan, Aokigahara woodland is considered to be one of the most natural areas around Mount Fuji and a core area in the conservation of the biodiversity of Mount Fuji. We chose butterflies as an indicator species of biodiversity and examined six communities in and around the woodland in 2000 using transect counts to examine and search for diversity and rarity hotspots and their associated landscapes. The results showed that butterfly species richness and species diversities H 1/ were significantly higher in forest-edge sites than in forest-interior and/or open-land sites, and variation in the total number of species among these three landscape types was well accounted for by ecologically specialist species, such as landscape specifics, oligovoltines, narrow diet feeders and low-density species. Thus, the species regarded as vulnerable to extinction, including Red List species, were observed more often in forest-edge sites than in forest-interior and/or open-land sites. As a result, in the study area, diversity and rarity hotspots were found in forest-edge landscapes. The reasons why butterfly diversity and rarity hotspots were established in forest-edge landscapes were analyzed and interpreted from several points of view, including disturbance level, landscape elements and plant species richness. From these results, and the fact that some species were confined to forest-interior sites, we conclude that it is very important to conserve and manage forest-edge habitats (considered to be semi-natural) as well as forest-interior habitats (considered to be the most natural) to maintain the diversity of butterfly communities and preserve the various types of threatened species in and around the Aokigahara woodland.     

Detection of 1270 nm emission from singlet oxygen and photocytotoxic property of sugar-pendant 60 fullerenes

Sugar-pendant [60] fullerene derivatives have been prepared from carbohydrate-linked azides 1a-e. Both monosugar (4a-e) and bissugar derivatives (5a-e) produce singlet oxygen ((1)O(2)) under laser irradiation (355 nm) proved by the direct observation of (1)O(2) emission at 1270 nm. Monosugar derivatives exhibit photocytotoxicity varying by the attached sugar molecule.     

Effect of nucleic acid binding on the triplet state properties of tetrapeptides containing tryptophan and 6-methyltryptophan: a study by phosphorescence and ODMR spectroscopy

Complexes of four peptides [KWGK, KGWK, K(6MeW)GK, KG(6MeW)K] with the nucleic acids [poly(A), poly(C), poly(U), poly(I), and rG(8)] have been investigated by phosphorescence and optically detected magnetic resonance (ODMR) spectroscopy. The intrinsic spectroscopic probes used in these studies are tryptophan (W) and 6-methyltryptophan (6MeW). Binding to the nucleic acids results in a red-shift of the phosphorescence 0,0-band (delta E(0,0)) of the aromatic residue as well as a reduction of its zero-field splitting parameter (delta D). Results are compared with earlier studies of the HIV-1 nucleocapsid protein, NCp7, that contains a single tryptophan residue (Trp37) within a retroviral zinc finger sequence. Binding of poly(A) or poly(U) to the tetrapeptides induces larger delta E(0,0) and delta D than when bound to NCp7, indicating stronger stacking interactions. Poly(I), on the other hand, produces larger shifts in Trp37 of NCp7. Binding of rG(8) produces sequence-dependent effects in the peptides. When bound to NCp7, but in contrast with tetrapeptide binding, nucleic acids produce large changes in triplet state kinetics consistent with enhanced spin-orbit coupling. These results are discussed in terms of three limiting types of tryptophan-base interaction: intercalation, aromatic stacking, and edge-on interaction. These should have differing effects on the properties of the triplet state.     

PDGF-BB enhances alpha1beta1 integrin-mediated activation of the ERK/AP-1 pathway involved in collagen matrix remodeling by rat mesangial cells

Platelet-derived growth factor-BB (PDGF-BB) has been implicated in the pathogenesis of progressive glomerulonephritis (GN). Previous studies have reported that PDGF-BB stimulates mesangial cells (MCs)-induced collagen matrix remodeling through enhancement of alpha1beta1 integrin-dependent migratory activity. To determine the cell signaling pathway responsible for abnormal MC-related mesangial matrix remodeling in progressive GN, we studied the involvement of the extracellular signal-regulated kinase (ERK)/activator protein-1 (AP-1) pathway in PDGF-BB-enhanced collagen gel contraction. Western blotting and gel shift assay revealed that MC-induced gel contraction resulted in ERK activation in parallel with that of AP-1 binding, peaking at 4 h and lasting at least for 24 h. Application of the MEK inhibitor, U0126, and the c-jun/AP-1 inhibitor, curcumin, inhibited gel contraction and AP-1 activity, respectively, dose dependently. PDGF-BB enhanced not only gel contraction but ERK phosphorylation and AP-1 activity by MCs. Marked inhibitory effects on PDGF-BB-induced gel contraction and ERK/AP-1 activity were observed in the presence of either function blocking anti-alpha1- or anti-beta1-integrin antibody or U0126. Consistently, AP-1-inactive MCs expressing a dominant-negative mutant of c-jun showed a significant decrease of PDGF-BB-induced gel contraction as compared with mock-transfected MCs. Finally, migration assay showed that ERK/AP-1 activity is required for PDGF-BB-stimulated alpha1beta1 integrin-dependent MC migration to collagen I. These results indicated that PDGF-BB enhances alpha1beta1 integrin-mediated collagen matrix reorganization through the activation of the ERK/AP-1 pathway that is crucial for MC migration. We conclude that the ERK/AP-1 pathway plays an important role in PDGF-BB-induced alpha1beta1 integrin-dependent collagen matrix remodeling; therefore, the inhibition of its pathway may provide a novel approach to regulate abnormal collagen matrix remodeling in progressive GN.     

Co-utilization of Bacillus subtilis and Flutolanil in controlling damping-off of tomato caused by Rhizoctonia solani

Damping-off of tomato caused by Rhizoctonia solani was controlled in a pot test using the biological agent, Bacillus subtilis RB14-C, and the chemical pesticide, Flutolanil. The co-utilization of B. subtilis RB14-C, and Flutolanil decreased the amount of Flutolanil used from 375 g/pot when Flutolanil was used alone to 94 g/pot, while exerting the same effect of reducing disease occurrence.