Biochemical Characterization of Secreted Proteases During Growth in Tetrahymena pyriformis WH-14: Comparison of Extracellular with Intracellular Proteases

Tetrahymena pyriformis strain WH-14 secreted large quantities of intracellular proteases into its culture medium during growth. Extracellular enzymes were purified to homogeneity from cell-free medium by ammonium sulfate precipitation, CM-Sephadex column chromatography, gel filtration, and DEAE-cellulose column chromatography. The DEAE-cellulose eluates were separated into four peaks (P-I, P-II, P-III, and P-IV), each of which exhibited a different specific activity toward azocasein and α-N-benzoyl-DL-arginine-ρ-nitroanilide (Bz-Arg-Nan). These four forms of the protease showed similarity in amino acid composition, molecular weight (21,000–24,000), and antigenic reactivity. They had pH optima at neutral range. P-I showed the highest specificity to azocasein whereas P-IV was most effective toward the synthetic substrates. The Km values for hydrolysis of Bz-Arg-Nan were 2.4, 1.6, 1.3, and 1.4 mM for P-I, P-II. P-III, and P-IV, respectively, and the corresponding Kcat/Km values were 5.0, 9.4, 28.5, and 114.3 S^-1.M^-1. These properties of secreted proteases were compared with those of intracellular proteases purified by the same procedure except for the initial Triton X-100 extraction. There were similarities in specific activity toward two substrates, molecular weight, Km, pH optima, and antigenic reactivity between the proteases from two sources, providing evidence that the intracellular proteases may be secreted into the extracellular medium without modification.     

Structure of Major Surface Determinants and DNA Diagnosis of Pneumocystis carinii

Pneumocystis carinii is a pathogen which, causes fatal pneumonia in patients with the acquired immune deficiency syndrome (AIDS). To facilitate the basic study of P. carinii , we have analyzed its major surface proteins by both immunochemical and biochemical methods. The major protein components of both cysts and trophozoites are a group of proteins called "P115" with apparent masses of 105–120 kd. It includes 6 isoelcclric variants. A monoclonal antibody raised against cysts recognizes all 6 variants and reacts with epitopes located in the cell wall indicating that P115 is an immunorcactive surface component. The isoelectric variants contain identical or closely related protein components and they are mannose-rich glycoproteins. The isoelectric variation may be due primarily to differences in glycosylation. The majority of sera from humans with diagnosed pneumocystosis that were tested reacted strongly with the P115 proteins. To develop probes for DNA diagnosis and to facilitate molecular studies, a genomic DNA library of P. carinii has been constructed. Some of these clones were used for DNA hybridization analysis of rat and human lungs.     

Immune-Enhancing Activity Screening on Extracts from Two Crickets, Gryllus bimaculatus and Teleogryllus emma

This study was performed to explore novel and valuable uses of insect resources, important subjects of the natural compound used in bio‐industries. The whole bodies of two crickets, Gryllus bimaculatus and Teleogryllus emma, selected from medicinal insect species, were carefully ground and treated with 80% EtOH. The insect extracts were solubilized and separated by hexane, butanol, and D.W according to their polarities. Three types of extracts, a D.W fraction (G1) and a boiling extract (G2) of an introduced cricket, G. bimaculatus, and a D.W fraction (T1) of a Korean local cricket, T. emma, were prepared to assay immune stimulating activity of cricket originated compounds. The all of three treated cricket extracts showed to increase IL‐4, IFN‐, and TNF‐α. Among those extract, extract G2, boiled extract from G. bimaculatus, was the best immune–enhancing fraction. The results of this study could be fundamental information for further works to use insects as natural resources having plenty of potentials and varieties.     

Molecular phylogeography reveals an antitropical distribution and local diversification of Solenogyne (Asteraceae) in the Ryukyu Archipelago of Japan and Australia

An antitropical distribution represents an intriguing disjunction, in which a given species or sister lineages occupy regions north and south of the tropics but are absent from the intervening areas. Solenogyne mikadoi endemic to the Ryukyu Archipelago is regarded as an Australian element. Testing the phylogenetic relationship with Australian congeners and discussing the onset timing and causes of the disjunction would potentially enhance the understanding of antitropical distribution. A nuclear ribosomal DNA phylogeny was reconstructed using Bayesian and most parsimonious criteria with allied genera. Solenogyne was monophyletic and clustered with Lagenophora huegelii endemic to Australia, indicating the antitropical distribution and Australian origin of Solenogyne. Multispecies coalescent analysis based on nuclear ribosomal DNA and chloroplast DNA indicated the divergence of S. mikadoi and Australian congeners in the Plio‐Pleistocene. Phylogenetic network analyses suggested that the ancestral lineage of S. mikadoi first colonized the southernmost island in the archipelago and then dispersed northward. The migration to the archipelago likely followed the flourishing of Solenogyne in open vegetation communities that radiated in south‐eastern Australia during the late Pliocene. This disjunction might arise through long‐distance dispersal across the tropics or, alternatively, through extinction in the tropics as a result of unsuitably high temperatures during climate oscillation and/or competitions from diverse tropical flora surviving since the early Tertiary. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 197–217.     

Relationship between fern development and CAM in Pyrrosia piloselloides (L.) Price

Pyrrosia piloselloides (L.) Price is a constitutive CAM plant in the sporophytic phase of its life-cycle. Newly developed sporophytes, still attached to the gametophytes, showed signs of CAM expression in terms of diurnal changes in titratable acidity of the tissues. The gametophytes did not exhibit CAM. This revised version was published online in June 2006 with corrections to the Cover Date.     

Upregulation of hemolymph and tissue ferritin by dietary HgCl2 in the wax moth Galleria mellonella

The effect of Hg treatment on hemolymph and tissue ferritin in the wax moth Galleria mellonella was examined by western blotting. At 48 h after feeding HgCl₂, the level of hemolymph ferritin increased approximately 1.8‐fold over that of control insects that were not fed HgCl₂, while there was a small increase in tissue ferritin. Time series experiments showed that tissue ferritin had a typically saturated pattern, with a maximum level from 24 to 72 h, although it decreased 12 h following HgCl₂ feeding, while hemolymph ferritin first decreased but subsequently increased. Tissue ferritin in the fat body, gut and Malpighian tubules, the main tissues of ferritin expression, was upregulated over time following treatment with Hg, and in particular, tissue ferritin in the gut increased by a large amount at 12–48 h. The results suggest that in G. mellonella, the ferritin‐inducible mechanisms following treatment with HgCl₂ are different for hemolymph and tissue ferritin, as are their biochemical properties.     

The biofuel potential of municipal solid waste

The world in the 21st century is facing a dual crisis of increasing waste and global climate change. Substituting fossil fuels with waste biomass‐derived cellulosic ethanol is a promising strategy to simultaneously meet part of our energy needs, mitigate greenhouse gas (GHG) emissions, and manage municipal solid waste (MSW). However, the global potential of MSW as an energy source is as yet unquantified. Here, we report increasing trends of MSW generation, and waste biomass‐derived cellulosic ethanol potentials in relation to socio‐economic development across 173 countries, and show that globally, up to 82.9 billion litres of waste paper‐derived cellulosic ethanol can be produced worldwide, replacing 5.36% of gasoline consumption, with accompanying GHG emissions savings of between 29.2% and 86.1%.     

韩国与中国东北地区松鼠线粒体DNA控制区的序列变异

为了研究松鼠东北亚种(Sciurus vulgaris manchuricusThomas)不同种群的序列变异水平并进一步确定分类地位,我们分析了韩国5个地点和中国东北2个地点的松鼠标本的线粒体DNA控制区的全序列(1 058 bp)。39个韩国松鼠标本显示出21种单倍型,这些单倍型间的平均Tamura-Nei距离为1·0%; 24个中国松鼠标本显示21种单倍型,单倍型间的平均Tamura-Nei距离为1·4% (1 058 bp的全序列中发生变异的位点有119个,占11·2%)。韩国松鼠和中国松鼠间的平均距离为1·3%。并且韩国和中国松鼠的所有42个单倍型形成了一个单系分支,Fst值为0·04,表明在两个国家的松鼠间没有发生遗传分化。因此,序列分析的分子生物学的结果支持现行的分类,即来自韩国的朝鲜亚种(S·v·coreae)是中国北部地区松鼠东北亚种(S·v·manchuricus)的同物异名。这还需要进一步对北朝鲜和中国东北其它地区更多标本的分子和形态学分析来验证这一结论。     

Effects of Cricket Extract on Lipid Metabolism and Body Fat Content in High- Fat Diet Fed Rats

The effect of cricket extract on high‐fat diet fed rat was observed. It was shown that cricket extract prevented the increment of body weight by high‐fat diet. The extract also decreased the value of AST in liver. The most significant effect of the extract was shown on lipid metabolism. The contents of total lipid and total cholesterol in liver and feces were reduced by the extract on dose‐dependent. These statistically significant results were clear in 3% extract treated group (HFD3) while were slight in 1% extract treated group (HFD1). The same result was also shown in body fat content.     

水稻低温发芽势的遗传及数量性状基因座分析

利用籼粳交“密阳23／吉冷1号”的F2：3代200个家系作为作图群体,在14℃条件下鉴定萌发7d、11d、14d和17d时低温发芽势,并利用由SSR标记构建的分子连锁图谱为基础,对不同萌发阶段的低温发芽势进行了数量性状基因座（QTLs）检测,同时进行了低温发芽势与其他耐冷性状间的相关分析。结果表明,萌发7d时低温发芽势及其低温反应指数呈现向低发芽势和低的低温反应指数的偏态分布,而萌发11d、14d和17d时低温发芽势及其低温反应指数均呈现接近正态的单峰连续分布。萌发14d时低温发芽势与其他耐冷性的相关性较萌发7d、11d和17d时低温发芽势更为密切,与芽期耐冷性、幼苗期耐冷性、低温下幼苗生长能力和孕穗期耐冷性表现为显著或极显著的正相关。位于第2染色体RM29-RM262区间的qLVG2和第7染色体RM336-RM118区间的qLVG7-2、qCIVG7-2在萌发11d、14d和17d时均检测到;位于RM29-RM262区间的qCIVG2在萌发11d和14d时均检测到,并对表型变异的贡献率随着萌发进程而逐渐增加。与低温发芽势相关的qLVG2贡献率从6．9％增加到14.2％,qLVG7-2贡献率从9．9％增加到11．2％,而与发芽势的低温反应指数相关的qCIVG2贡献率从6.3％增加到9．0％,qCIVG7-2贡献率从8．3％增加12．9％。这些QTL的增效等位基因均来自强耐冷亲本吉冷1号,基因作用方式主要为部分显性。