Properties of enzymes from bacteria grown in the 70–100°C range

In chemostat cultures of Bacillus caldolyticus, adaptation in a single step from 70–100°C was followed under aerobic and oxygen-limited conditions and was found to proceed more smoothly under the latter circumstances. Variations of the medium (e.g. yeast extract or silicate concentrations) showed that growth at 100°C is in all respects similar to that of cultures at moderate temperatures.Enzyme preparations derived from cultures at 5°C intervals between 70 and 100°C were used to determine the temperature range. For all nine enzymes tested, the optimum temperature was found to be 67°C; the latter was independent of the growth temperature. Differences were found, however, with respect to the maximum temperature of individual enzymes, and three groups, with maxima between 70 and 80°C, 80 and 90°C and 90 and 100°C can be distinguished. Again, there was no correlation with the growth temperature.Stability experiments also revealed that enzymes from the same organism can have different thermal properties: Some were found to be quite thermolabile (e.g. the pyruvate kinase), while others (e.g. hexokinase and glutamate-pyruvate transaminase) exhibited a high thermostability. These properties were not related to the growth temperature within the 70–100°C range, too.Six of the enzymes tested could be stabilized by their respective substrates, but the degree of protection varied for individual enzymes. Three enzymes (acetate kinase, glutamate dehydrogenase and myokinase) could not be stabilized by their substrates.Comparative experiments with the hexokinase suggested, that the thermal integrity of the enzymes is better protected within the cell as compared to the stability of the enzyme preparations.Abbreviations used AK acetate kinase - Ala-DH alanine dehydrogenase - Ald aldolase - GIDH glutamate dehydrogenase - G6P-DH glucose-6-phosphate dehydrogenase - GTP glutamate-pyruvate transaminase - HK hexokinase - ICDH isocitrate dehydrogenase - MK myokinase - PK pyruvate kinase     

乙型脑炎病毒NS2A-C60A位点突变对其生物学特性影响研究*

目的:旨在探索Ⅰ型日本乙型脑炎病毒传代致弱后基因组突变NS2A-C60A对乙脑病毒生物学特性的影响。方法:首先通过对传代致弱及原始乙脑毒株基因组序列进行测序比对、结构预测分析并利用Western blotting(WB)确定了目标研究位点NS2A-C60A;然后使用反向遗传定点突变技术构建拯救了包含NS2A-C60A单点突变的病毒株;最后利用噬斑形态观察、生长曲线、双萤光素酶分析,WB以及炎性因子检测和动物实验研究了该单点突变对于乙脑病毒生物学特性的影响。结果:首次研究发现Ⅰ型乙脑病毒传代致弱会导致NS1'蛋白表达的显著下降以及可能的相关位点NS2A-C60A,并成功拯救获得了NS2A-C60A单点突变毒株rJEV-C60A,研究发现NS2A-C60A突变对乙脑病毒的生长特性及噬斑形成没有显著影响,但是能够显著降低乙脑病毒NS1'蛋白的表达,并且该位点突变能够轻微阻碍乙脑病毒对细胞炎性因子表达的抑制,动物实验结果显示NS2A-C60A点突变病毒与原毒株具有相似的神经毒力,说明该位点突变不是影响乙脑病毒毒力致弱的关键位点。结论:新发现的NS2A-C60A位点突变能够显著减少乙脑病毒NS1'蛋白的表达,但是对其增殖、诱导炎症及神经毒力等生物学特性没有显著影响。     

木犀科植物叶绿体基因组结构特征和系统发育关系

木犀科11属19个种叶绿体基因组的一般特征和变异特征的比较分析显示, 结果表明, 该科叶绿体基因组大小为154-165 kb, 其差异主要是大单拷贝(LSC)长度的差异所致。Jasminum属3个物种的叶绿体基因组长度与其余物种有较大差异, 该属clpP基因内含子和accD基因丢失。共线性分析表明, Jasminum属3个物种多个基因出现基因重排现象, 倒位可能是重排的主要原因。Jasminum属在IRb/SSC和SSC/IRa边界的基因均与其它物种不同; 重复序列与SSR数量检测结果表明, Jasminum属与其余物种在数量及重复长度上差异较大。基于CDS数据构建的系统发育树表明, Abeliophyllum distichum和Forsythia suspensa为木犀科中较早分化的类群。     

Spoil characteristics and vegetation development of an age series of mine spoils in a dry tropical environment

A series of coal mine spoils (5, 10, 12, 16 and 20-yr old) in a dry tropical environment was sampled to assess the changes with time in spoil characteristics, species composition and plant biomass. Coarse fragments (>2 mm) decreased with age of mine spoil while the proportion of 0.2–0.1 mm particles increased. Total soil N, mineral N, NaHCO₃-extractable Pi, and exchangeable K increased with age of mine spoil and these parameters were lower in mine spoils than native forest soil even after 20 years of succession. Exchangeable Na decreased with age of mine spoil and in 20-yr old spoil it was higher than native forest soil. Plant community composition changed with age. Only a few species participated in community formation. Species richness increased with age, while evenness and species diversity declined from 5-yr old to 16-yr old community with an increase in the 20-yr old community. A reverse trend occurred for concentration of dominance. Area-weighted shoot and root biomass of other species increased with the age of the mine spoil while that of Xanthium strumarium patches declined with age. Data collected on spoil features, microbial C, N and P, and shoot and root biomass when subjected to Discriminant Analysis indicated a continued profound effect of age. 10 and 12-yr old mine spoils were closer to each other, and 5 and 20-yr old spoils were farthest apart.     

Studies on a Morphologically Distinct Colchicine-Resistant Variant of Entamoeba sp.*

Colchicine has a temperature-dependent cytotoxic effect on Entamoeba sp. (Laredo isolate) that is most apparent when the drug is applied during the initiation of cultures at a concentration of 7.5 mM or higher. Continued transfer of cultures in medium containing progressively increasing concentrations of colchicine has resulted in a variant that grows prolifically in the presence of colchicine (7.5 mM) with a generation time comparable to that of the parent stock. Comparison of a number of parameters of the 2 variants revealed that colchicine resistance was accompanied by a change in cell shape, a reduced membrane permeability, which could partially be overcome by the addition of dimethyl sulfoxide (DMSO), and a reduced tolerance to osmotic stress. However, the parent strain and resistant variant were equally susceptible to cycloheximide and puromycin suggesting that the acquired colchicine resistance may not be explained on the basis of an entirely unspecific generalized reduced ability for drug uptake. Colchicine resistance and altered structure were found to be stable over a long period of time. The possible interdependence of these 2 parameters and their relation to cell motility in Entamoeba sp. are discussed.     

An optimal dietary non-phytate phosphorus level of broilers fed a conventional corn–soybean meal diet from 4 to 6 weeks of age

It is imperative to evaluate precise nutrient requirements of animals in order to optimize productivity and minimize feed cost and nutrient excretions. The current non-phytate phosphorus (NPP) recommendation for broilers is based on the papers published 30 years ago. However, today’s commercial birds are quite different from those before 30 years. Therefore, the present experiment was conducted with growing male broiler chickens to evaluate an optimal dietary NPP level of broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age. The 1-day-old chicks were fed corn–soybean meal diet containing 0.39% NPP from 1 to 3 weeks of age. At 22 days of age, 360 birds were selected and randomly allotted by BW to one of 10 dietary treatments with six replicate cages of six birds per cage for each treatment. Birds were fed the P-unsupplemented corn–soybean meal basal diet and the basal diet supplemented with inorganic P as CaHPO₄·H₂O ranging from 0.00% to 0.45% with 0.05% increment from 4 to 6 weeks of age. The dietary NPP levels were 0.09%, 0.14%, 0.20%, 0.24%, 0.30%, 0.34%, 0.38%, 0.45%, 0.49% and 0.54%, respectively, and the dietary Ca level was fixed at 0.90% for all treatments. The results showed that average daily gain, serum inorganic P concentration, tibia bone strength, tibia ash percentage and P percentage, tibia bone mineral content (BMC) and density (BMD), middle toe ash percentage and P percentage, middle toe BMC, total body BMC and BMD were affected (P<0.0001) by dietary NPP level, and increased linearly (P<0.0001) and quadratically (P<0.003) as dietary NPP levels increased. Optimal dietary NPP levels estimated based on fitted broken-line models (P<0.0001) of the above indices are 0.21%, 0.29%, 0.29%, 0.29%, 0.29%, 0.31%, 0.29%, 0.30%, 0.27%, 0.29% and 0.28%, respectively. It is suggested that the total body BMC and BMD, and middle toe ash P and BMC might be new, sensitive and non-invasive criteria to evaluate the dietary NPP requirements of broilers. The optimal dietary NPP level would be 0.31% for broiler chickens fed a conventional corn–soybean meal diet from 4 to 6 weeks of age.     

Individually mark–mass release–resight study elucidates effects of patch characteristics and distance on host patch location by an insect herbivore

1. How organisms locate their hosts is of fundamental importance in a variety of basic and applied ecological fields, including population dynamics, invasive species management and biological control. However, tracking movement of small organisms, such as insects, poses significant logistical challenges. 2. Mass‐release and individual–mark–recapture techniques were combined in an individually mark–mass release–resight (IMMRR) approach to track the movement of over 2000 adult insects in an economically important plant–herbivore system. Despite its widespread use for the biological control of the invasive thistle Carduus nutans, the host‐finding behaviour of the thistle head weevil Rhinocyllus conicus has not previously been studied. Insects were released at different distances from a mosaic of artificially created host patches with different areas and number of plants to assess the ecological determinants of patch finding. 3. The study was able to characterize the within‐season dispersal abilities and between‐patch movement patterns of R. conicus. Weevils found host plant patches over 900 m away. Large patches, with tall plants, situated close to the nearest release point had the highest first R. conicus resights. Patch area and plant density had no effect on the number of weevils resighted per plant; however, R. conicus individuals were more likely to disperse out of small patches and into large patches. 4. By understanding how R. conicus locates host patches of C. nutans, management activities for the control of this invasive thistle can be better informed. A deeper mechanistic understanding of host location will also improve prediction of coupled plant–herbivore spatial dynamics in general.     

A Turkish family with Nance-Horan syndrome due to a novel mutation

Nance-Horan Syndrome (NHS) is a rare X-linked syndrome characterized by congenital cataract which leads to profound vision loss, characteristic dysmorphic features and specific dental anomalies. Microcornea, microphthalmia and mild or moderate mental retardation may accompany these features. Heterozygous females often manifest similarly but with less severe features than affected males. We describe two brothers who have the NHS phenotype and their carrier mother who had microcornea but not cataract. We identified a previously unreported frameshift mutation (c.558insA) in exon 1 of the NHS gene in these patients and their mother which is predicted to result in the incorporation of 11 aberrant amino acids prior to a stop codon (p.E186Efs11X). We also discussed genotype–phenotype correlation according to relevant literature.     

Forest-edge utilization by carnivores in relation to local and landscape habitat characteristics in central European farmland

Rapid changes in agricultural landscape structure and composition affect many different farmland biotas, including carnivores, which are a key element of ecosystem stability, yet little is known about their distribution and habitat use. In this study, we evaluated how habitat characteristics on two different spatial scales (local and landscape scale) affected the forest-edge utilization by small and medium-sized carnivores in fragmented central European farmland. Based on an indirect method for detecting carnivores (scent stations), we sampled 212 forest fragments of different sizes (1–7864 ha) during April to May from 2006 to 2009. Our results indicate that carnivore utilization of forest-edge habitats was driven by landscape rather than local characteristics even though the overall extent of explained variation was small. The most important factors that determined response of the carnivore community were the area of farmland and that of urban land on a landscape scale. The corridor connectivity between small forest fragments and other spatial elements played a crucial role in the occurrence of red fox. Our results suggest that comprehensive studies on multi-species carnivore assemblage using scent station might be useful in evaluating species-specific response to habitat characteristics, especially if large numbers of stations visited by carnivores are available.