Diversity of fimbrillin among Porphyromonas gulae clinical isolates from Japanese dogs

Porphyromonas gulae, a gram-negative black-pigmented anaerobe, is a pathogen for periodontitis in dogs. An approximately 41-kDa fimbrial subunit protein (FimA) encoded by fimA is regarded as associated with periodontitis. In the present study, the fimA genes of 17 P. gulae strains were sequenced, and classified into two major types. The generation of phylogenetic trees based on the deduced amino acid sequence of FimA of P. gulae strains along with sequences from several strains of Porphyromonas gingivalis, a major cause of human periodontitis, revealed that the two types of FimA (types A and B) of P. gulae were similar to type I FimA and types II and III FimA of P. gingivalis, respectively. A PCR system for classification was established based on differences in the nucleotide sequences of the fimA genes. Analysis of 115 P. gulae-positive oral swab specimens from dogs revealed that 42.6%, 22.6%, and 26.1% of them contained type A, type B, and both type A and B fimA genes, respectively. Experiments with a mouse abscess model demonstrated that the strains with type B fimA caused significantly greater systemic inflammation than those with type A. These results suggest that the FimA proteins of P. gulae are diverse with two major types and that strains with type B fimA could be more virulent.     

Detection of a quantitative trait locus for intramuscular fat accumulation using the OLETF rat

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type 2 diabetes. We showed that the OLETF rat exhibits higher levels of intramuscular fat content in Musculus longissimus as compared to the Fischer-344 (F344) rat. Our investigation was designed to identify quantitative trait loci (QTLs) contributing to the increased levels of intramuscular fat content by performing a whole-genome search using 108 F2 intercross obtained by mating the OLETF and the F344 rats. We identified one QTL responsible for intramuscular fat accumulation on rat chromosome 1 with a maximum lod score of 3.4, which accounts for 5% of the total variance. As expected, the OLETF allele corresponds to the increased levels of intramuscular fat content.     

Stimulatory effect of cold acclimation on skeletal muscle branched-chain α-keto acid dehydrogenase in rats

The effects of cold-acclimation (3 weeks at 4°C) on the actual activities of branched-chain α-keto acid dehydrogenase (BCKDH) and pyruvate dehydrogenase (PDH) complexes in the skeletal muscle and liver of normal 6-week-old rats were studied. The activities of BCKDH and PDH complexes were assayed using [1–¹⁴C] α-ketoisocaproate and [1-¹⁴C]pyruvate, respectively. Cold-acclimation markedly stimulated the activity of BCKDH, and slightly but significantly increased the activities of PDH and citrate synthase (CS) in the skeletal muscle, but did not alter PDH, BCKDH or CS activities in the liver. The increase in muscle BCKDH activity (control rats, 1.45 ± 0.54 mU/g wet wt; cold-acclimated rats, 2.54 ± 0.50 mU/g wet wt; 75% increase: P  = 0.001) was much greater than the increases in PDH activity (84 ± 16 mU/g wet wt and 111 ± 25 mU/g wet wt; 32% increase: P  = 0.020) or CS activities (27 ± 3 μmol/min per g wet wt and 31 ± 2 μmol/min per g wet wt; 14% increase: P  = 0.046). These results suggest that, unlike PDH, BCKDH is not directly associated with the mitochondrial oxidative activity of the skeletal muscle of cold-acclimated rats. This study provides the first evidence that BCKDH in skeletal muscle responds selectively to cold-acclimation.     

Mechanisms responsible for reduced cardiotoxicity of mitoxantrone compared to doxorubicin examined in isolated guinea-pig heart preparations

We reported previously that doxorubicin, an anticancer agent that has an anthracycline structure, alters Ca2+ releasing and uptake mechanisms in the sarcoplasmic reticulum of myocardial cells. These effects of doxorubicin are apparently related to its cardiotoxicity. Mitoxantrone is a similar anticancer agent with an anthracenedion structure that has been shown to be significantly less cardiotoxic. In the present study, the effects of mitoxantrone on the functions of the sarcoplasmic reticulum were examined in isolated muscle preparations obtained from the guinea-pig heart. In electrically-stimulated left atrial muscle preparations, incubation in vitro for 4 hr with 30 or 100 microM mitoxantrone significantly prolonged the time to the peak of twitch tension, markedly increased the developed tension observed at lower stimulation frequencies, thereby attenuating the slope of positive force-frequency relationships, and increased the postrest contraction observed after a 60-sec quiescent period. In myocytes isolated from ventricular muscles, 30 microM mitoxantrone increased the peak and the size of intracellular Ca2+ concentrations ([Ca2+] i), and prolonged the time to peak [Ca2+]i. In skinned muscle fiber preparations obtained from the left ventricular muscle, 30 muM mitoxantrone significantly increased the caffeine-induced contraction without affecting the Ca2+ sensitivity of contractile proteins. These results suggest that mitoxantrone enhances Ca2+ release from the sarcoplasmic reticulum in isolated atrial muscle preparations obtained from the guinea-pig heart. Apparent enhancement of the sarcoplasmic reticulum functions, in contrast to anthracyclines that has been shown to suppress these functions, seems to explain the relative lack of marked cardiotoxicity of mitoxantrone.     

Utilization of flow cytometry for festulolium breeding (Lolium multiflorum (2x) × Festuca arundinacea (6x))

Festulolium is a hybrid between Festuca and Lolium species that has valuable agronomic traits from both grass species. The purpose of our breeding program is to produce hexaploid festulolium that introduces tolerance to summer depression into Italian ryegrass (Lolium multiflorum) by crossing it with tall fescue (Festuca arundinacea). However, we found the DNA ploidy of hexaploids was not stable and was reduced in successive generations. We aimed to find out how to obtain stable high-ploidy festulolium. F1 hybrids of L. multiflorum and F. arundinacea were produced. The F3 generation was produced from putative hexaploid F2 individuals by open pollination. The F4 to F6 generations were obtained by polycrossing. The DNA ploidy levels of F2 to F6 individuals were estimated by flow cytometry. Cytological characteristics of the F5 and F6 individuals were investigated by FISH and GISH. The DNA ploidy level of hexaploid festulolium was reduced and stabilized at almost the same level as a tetraploid. Seed fertility was inversely correlated with an increase in ploidy level. GISH revealed no preferential Lolium transmission. FISH with a telomere probe revealed that counting the exact number of chromosomes in festulolium was difficult. DNA ploidy level was strongly correlated with the number of chromosomes.     

Synergistic, additive, and antagonistic effects of food mixtures on total antioxidant capacities

Different foods possess different bioactive compounds with varied antioxidant capacities. When foods are consumed together, the total antioxidant capacity of food mixtures may be modified via synergistic, additive, or antagonistic interactions among these components, which may in turn alter their physiological impacts. The main objective of this study was to investigate these interactions and identify any synergistic combinations. Eleven foods from three categories, including fruits (raspberry, blackberry, and apple), vegetables (broccoli, tomato, mushroom, and purple cauliflower), and legumes (soybean, adzuki bean, red kidney bean, and black bean) were combined in pairs. Four assays (total phenolic content, ferric reducing antioxidant power, 2,2-diphenyl-1-picrylhydrazyl, radical scavenging capacity, and oxygen radical absorbance capacity) were used to evaluate the antioxidant capacities of individual foods and their combinations. The results indicated that within the same food category, 13, 68, and 21% of the combinations produced synergistic, additive, and antagonistic interactions, respectively, while the combinations produced 21, 54, and 25% synergistic, additive, and antagonistic effects, respectively, across food categories. Combining specific foods across categories (e.g., fruit and legume) was more likely to result in synergistic antioxidant capacity than combinations within a food group. Combining raspberry and adzuki bean extracts demonstrated synergistic interactions in all four chemical-based assays. Compositional changes did not seem to have occurred in the mixture. Results in this study suggest the importance of strategically selecting foods or diets to maximum synergisms as well as to minimum antagonisms in antioxidant activity.     

Properties of soil humic substances in fractions obtained by sequential extraction with pyrophosphate solutions at different pHs

Abstract

Humic acids (HAs) obtained from four different soils by sequential extraction with pyrophosphate solutions at different pH values (i.e. at pH 3, 5, 7, 9, 11, and 13), were examined by ultraviolet and visible (UV-VIS) spectroscopy and elemental analysis. UV-VIS spectra from 230 to 700 nm were measured, and absorbance coefficients of 1% humic acid solutions at 600 nm (E ^1% ₆₀₀) and Δlog K were calculated. The shape of the spectra of humic acids depended on the soil types rather than on the pH values. The (E ^1% ₆₀₀) values were higher in the neutral region, and lower in the alkaline region, for each pH value they decreased in the following order: buried Andosol > Andosol > Histosol ≧ Cambisol. The Δlog K values for each pH value were higher in the following order: Histosol> Cambisol > buried Andosol≧Andosol. Although the linear correlation was found to be significant between the E ^1% ₆₀₀ and Δlog K values, the relation between these optical properties and pH values differed among the soil types. However, in the neutral region, the higher E ^1% ₆₀₀ values of humic acids from buried Andosol and Andosol and the lower Δlog K values of humic acids from Cambisol and Histosol suggest that the humification degree was higher in the neutral region. Data of elemental composition and atomic ratios were obtained for each humic acid. The H/C ratios in the four soils increased gradually with increasing pH values from 5 to 13, and those for each pH value were commonly higher in the following order: Cambisol> Histosol> Andosol> buried Andosol. The O/C and OIH ratios decreased with increasing pH values. The results of the H/C versus O/C diagram suggested that decarboxylated humic acids were extracted at the higher pH values. A highly significant correlation coefficient was obtained between the H/C and OIH ratios (r= ?0.891***). Results showing that the decrease of the pH values corresponded to the order of the humic acids in the correlation curve in the H/C versus O/H diagram suggest that the humic acids with a higher content of unsaturated bonds and a higher degree of oxidation were extracted at the lower pH values within the range from 5 to 13.     

马尾松造林密度试验研究

通过对马尾松4种造林密度的定位观测分析结果表明，不同造林密度对胸径、单株村积生长的影响达到显著性差异，对树高生长影响差异不显著，不同密度林分径级分布差异明显，密度为2500株／hm2和1250株／hm2的林分，其胸径≤6cm径阶的株数占57．2％和45．9％，≥8cm径阶的株数占42．8％和54．1％；而密度为10000株／hm2和5000株／hm2的林分，其胸径≤6cm径阶的株数占89．6％和76％，≥8cm径阶的株数占10．4％和24％；不同径级林分的合理密度通过胸径与树冠面积的相关关系，用回归直线方程y=a＋bx分别求得各胸径最低保留株数，作为间伐强度、间伐开始期和间伐重复期的参考依据。     

White Baron: A non-browning somaclonal variant of Danshakuimo (Irish Cobbler)

White Baron is a somaclonal variant variety resulting from the protoplast propagation of Danshakuimo, the dominant variety in the Japanese tablestock market since the beginning of this century. White Baron is an improvement over the parent clone in that its tubers do not brown after peeling whereas the tubers of the original variety brown rapidly after peeling. This commercially attractive improvement makes this variety well suited for the expanding market in Japan for pre-peeled potatoes.