Proteome analysis of a rat liver nuclear insoluble protein fraction and localization of a novel protein, ISP36, to compartments in the interchromatin space

A rat liver nuclear insoluble protein fraction was analyzed to investigate candidate proteins participating in nuclear architecture formation. Proteins were subjected to two-dimensional separation by reversed-phase HPLC in 60% formic acid and SDS/PAGE. The method produced good resolution of insoluble proteins. One hundred and thirty-eight proteins were separated, and 28 of these were identified. The identified proteins included one novel protein, seven known nuclear proteins and 12 known nuclear matrix proteins. The novel 36 kDa protein was further investigated for its subnuclear localization. The human ortholog of the protein was expressed in Escherichia coli and antibodies were raised against the recombinant protein. Exclusive localization of the protein to the nuclear insoluble protein fraction was confirmed by cell fractionation followed by immunoblotting. Immunostaining of mouse C3H cells suggested that the 36 kDa protein was a constituent of an insoluble macromolecular complex spread throughout the interchromatin space of the nucleus. The protein was designated 'interchromatin space protein of 36 kDa', ISP36.     

A genetic linkage map of azuki bean constructed with molecular and morphological markers using an interspecific population (Vigna angularis x V. nakashimae)

A genetic linkage map of azuki bean (Vigna angularis) was constructed with molecular and morphological markers using an F₂ population of an interspecific cross between azuki bean and its wild relative, V. nakashimae. In total, 132 markers (108 RAPD, 19 RFLP and five morphological markers) were mapped in 14 linkage groups covering 1250 cM; ten remained unlinked. The clusters of markers showing distorted segregation were found in linkage groups 2, 8 and 12. By comparing the azuki linkage map with those of mungbean and cowpea, using 20 RFLP common markers, some sets of the markers were found to belong to the same linkage groups of the respective maps, indicating that these linkage blocks are conserved among the three Vigna species. This map provides a tool for markerassisted selection and for studies of genome organization in Vigna species.     

Translocation of organic substances in sunflower II. Role of sugar phosphate in translocation of sugars

A single sunflower leaf was fed ¹⁴C-glucose for one min. Aftervarions periods, two sections of the plant stem above and belowthe fed leaf were cut into several sections and examined forradioactivity. Sucrose phophate was identified in the extracts after neutronirradiation of the chromatograms. A correlation was noted between the proportion of sucrose phosphatein the fed leaf and the rate of translocation from it. (Received May 22, 1978; )     

Effect of irrigation systems on temporal distribution of malaria vectors in semi-arid regions

Previous research models have used climate data to explain habitat conditions of Anopheles mosquitoes transmitting malaria parasites. Although they can estimate mosquito populations with sufficient accuracy in many areas, observational data show that there is a tendency to underestimate the active growth and reproduction period of mosquitoes in semi-arid agricultural regions. In this study, a new, modified model that includes irrigation as a factor was developed to predict the active growing period of mosquitoes more precisely than the base model for ecophysiological and climatological distribution of mosquito generations (ECD-mg). Five sites with complete sets of observational data were selected in semi-arid regions of India for the comparison. The active growing period of mosquitoes determined from the modified ECD-mg model that incorporated the irrigation factor was in agreement with the observational data, whereas the active growing period was underestimated by the previous ECD-mg model that did not incorporate irrigation. This suggests that anthropogenic changes in the water supply due to extensive irrigation can encourage the growth of Anopheles mosquitoes through the alteration of the natural water balance in their habitat. In addition, it was found that the irrigation systems not only enable the active growth of mosquitoes in dry seasons but also play an important role in stabilizing the growth in rainy seasons. Consequently, the irrigation systems could lengthen the annual growing period of Anopheles mosquitoes and increase the maximum generation number of mosquitoes in semi-arid subtropical regions.     

A new sand whiting, Sillago (Sillago) caudicula, from Oman, the Indian Ocean (Perciformes: Sillaginidae)

A new species of sand whiting, Sillago (Sillago) caudicula, is described based on four specimens collected from Oman, the Indian Ocean. It is easily distinguishable from its four known members of the subgenus Sillago by having a smaller head (29.0–30.1% in standard length), 23–24 soft anal fin rays, 35–36 total vertebrae, body depth at the origin of the second dorsal fin slightly deeper than that at the origin of the first dorsal fin, first and second hypurals fused (in adult) or narrowly separated (in young), third and fourth hypurals fused, and 11 dusky midlateral spots on the body.     

Heterogeneity in the conformation of valine in the elastin mimetic (LGGVG)6 as shown by solid-state 13C NMR SPEctroscopy

Elastin is an abundant protein found in vertebrates and is the source of elasticity in connective tissues and blood vessels. The repeating polypeptide sequences found in the hydrophobic domains of elastin have been the focus of many studies that attempt to understand the function of the native protein on a molecular scale. In this communication, the (LGGVG)6 elastin mimetic is characterized by solid-state 13C NMR spectroscopy. Through the use of a combination of a statistical analysis based on the Protein Data Bank, one-dimensional cross-polarization magic-angle-spinning NMR spectroscopy, and two-dimensional off-magic-angle-spinning spin-diffusion experiments, it is determined that this tandem repeat does not form a regular, highly ordered structure. Instead, like the poly(VPGVG) elastin mimetics, the valine has a twofold heterogeneity, although the conformations of these two populations differ from one peptide to the other.     

Resveratrol ameliorates myocardial damage by inducing vascular endothelial growth factor-angiogenesis and tyrosine kinase receptor Flk-1

In our study, resveratrol (polyphenol) has been identified as a very important stimulus/agent for the induction of new vessel growth. Occlusion of a main coronary depletes the blood supply to the myocardium and subsequently reduces cardiac function, which ultimately leads to heart failure. Progressive, chronic coronary artery occlusion has been shown to induce development of collateral arteries to re-establish and maintain blood flow to the myocardium at risk via the growth of new capillary vessels or angiogenesis. Studies from our laboratory, as well as from others, have already confirmed the protective role of collaterals against myocardial ischemia and cell death. We have successfully demonstrated in rat myocardial infarction (MI) model an effect of resveratrol on significant upregulation of the protein expression profiles of vascular endothelial growth factor (VEGF) and its tyrosine kinase receptor Flk-1, 3 wk after MI. Pretreatment with resveratrol also increased nitric-oxide synthase (inducible NOS and endothelial NOS) along with increased antiapoptotic and proangiogenic factors nuclear factor (NF)-kappaB and specificity protein (SP)-1. We also were able to demonstrate increased capillary density as well as improved left ventricular function by pharmacological preconditioning with resveratrol 3 wk after MI.     

Population genetic structure of Japanese wild soybean (Glycine soja) based on microsatellite variation

The research objectives were to determine aspects of the population dynamics relevant to effective monitoring of gene flow in the soybean crop complex in Japan. Using 20 microsatellite primers, 616 individuals from 77 wild soybean (Glycine soja) populations were analysed. All samples were of small seed size (< 0.03 g), were directly collected in the field and came from all parts of Japan where wild soybeans grow, except Hokkaido. Japanese wild soybean showed significant reduction in observed heterozygosity, low outcrossing rate (mean 3.4%) and strong genetic differentiation among populations. However, the individual assignment test revealed evidence of rare long-distance seed dispersal (> 10 km) events among populations, and spatial autocorrelation analysis revealed that populations within a radius of 100 km showed a close genetic relationship to one another. When analysis of graphical ordination was applied to compare the microsatellite variation of wild soybean with that of 53 widely grown Japanese varieties of cultivated soybean (Glycine max), the primary factor of genetic differentiation was based on differences between wild and cultivated soybeans and the secondary factor was geographical differentiation of wild soybean populations. Admixture analysis revealed that 6.8% of individuals appear to show introgression from cultivated soybeans. These results indicated that population genetic structure of Japanese wild soybean is (i) strongly affected by the founder effect due to seed dispersal and inbreeding strategy, (ii) generally well differentiated from cultivated soybean, but (iii) introgression from cultivated soybean occurs. The implications of the results for the release of transgenic soybeans where wild soybeans grow are discussed.     

Determination of casopitant and its three major metabolites in dog and rat plasma by positive ion liquid chromatography/tandem mass spectrometry

A sensitive, selective and quantitative method for the simultaneous determination of casopitant, a potent and selective antagonist of the human Neurokinin 1 (NK-1) receptor, and its three major metabolites M12, M13 and M31 was developed and validated in dog and rat plasma. Acetonitrile containing stable labeled internal standards for the four analytes was used to precipitate proteins in plasma. Chromatographic separation was obtained using a reversed phase column with multiple reaction monitoring turboionspray positive ion detection. The lower and upper limits of quantification for casopitant and its metabolites were 15 and 15,000 ng/mL, using a 50 μL of dog or rat plasma aliquot, respectively. The inter-day precision (relative standard deviation) and accuracy (relative error) in dog plasma, derived from the analysis of validation samples at 5 concentrations, ranged from 4.1% to 10.0% and −10.8% to 8.7%, respectively, for casopitant and its 3 major metabolites. The intra-day precision (relative standard deviation) and accuracy (relative error) in rat plasma, derived from the analysis of validation samples at 5 concentrations, ranged from 3.9% to 6.6% and −9.6% to 8.3%, respectively, for casopitant and its three metabolites. All analytes were found to be stable in analytical solutions for at least 43 days at 4 °C, in dog and rat plasma at room temperature for at least 24 h, at the storage temperature of −20 °C for at least 6 months, and following the action of three freeze–thaw cycles from −20 °C to room temperature. All analytes were also found to be stable in processed extracts at 4 °C for at least 72 h. This assay proved to be accurate, precise, fast and was used to support long-term toxicology studies in dog and rat.