Steady-state kinetics of the hypoxanthine phosphoribosyltransferase from Trypanosoma cruzi

The kinetic mechanism for the reaction catalyzed by the hypoxanthine phosphoribosyltransferase (HPRT) from Trypanosoma cruzi was analyzed to determine the feasibility of designing a parasite-specific mechanism-based inhibitor of this enzyme. The results show that the HPRT from T. cruzi follows an essentially ordered bi–bi reaction, and like its human counterpart also likely forms a dead end complex with purine substrates and the product pyrophosphate. Computational fitting of the kinetics data to multiple initial velocity equations gave results that are consistent with the dead end complex arising when the hypoxanthine- or guanine-bound form of the enzyme binds pyrophosphate rather than the phosphoribosylpyrophosphate substrate of the productive forward reaction. Limited proteolytic digestion was employed to provide additional support for formation of the dead end complex and to estimate the K_d values for substrates of both the forward and reverse reactions. Due to similarities with the kinetic mechanism of the human HPRT, the results reported here for the HPRT from T. cruzi indicate that the design of a mechanism-based inhibitor of the trypanosomal HPRT, that would not also inhibit the human enzyme, may be difficult. However, the results also show that a potent selective inhibitor of the trypanosomal HPRT might be achieved via the design of a bi-substrate type inhibitor that incorporates analogs of moieties for a purine base and pyrophosphate.     

Molecular Characterization of Tree Peony Germplasm Using Sequence-Related Amplified Polymorphism Markers

This study examined 63 tree peony specimens, consisting of 3 wild species and 63 cultivars, using sequence-related amplified polymorphism (SRAP) markers for the purpose of detecting genomic polymorphisms. Bulk DNA samples from each specimen were evaluated with 23 SRAP primer pairs. Among the 296 different amplicons, 262 were polymorphic. The maximum parsimony, neighbor-joining, and unweighted pair-group method using arithmetic average trees were largely in congruence. In the three trees, the wild species Paeonia ludlowii and P. delavayi formed separate clusters with strong bootstrap support, and P. ostii was closely related to all cultivars. The cultivars were divided into groups with various corresponding bootstrap values. The genetic similarity among the genotypes ranged from 0.02 to 0.73. These results demonstrate that SRAP markers are effective in detecting genomic polymorphisms in the tree peony and should be useful for linkage map construction and molecular marker assisted selection breeding. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Thirteen years of change in trees and lianas in a Gabonese rainforest

Inventories of trees and lianas were performed at a 13-year interval (1979–1992) in an evergreen forest in north-east Gabon. The number of trees decreased while their basal area and species richness increased. In contrast, both the richness and density of lianas decreased. Lianas with active climbing mechanisms were the main types to disappear.     

冷驯化对中缅树能量代谢的影响

在 5± 1℃条件下对中缅树鼠句进行冷驯化处理 ,测定其能量代谢。冷驯化 2 8d后 ,体重比对照组显著增加 7 3 3 % ;整体能值达到 3 0 47± 0 46kJ/g (N =8) ,比对照组增加4 98% ;摄入能比对照组增加 3 6 1 7% ;同化能比对照组增加 6 6 2 % ;生长能达到 6 98±0 5 3kJ/1 0 0g (N =7)体重·天 ,是对照组的 4 85倍 ;维持能比对照组增加 6 4 0 % ,达到3 5 2 96± 2 8 3 4kJ/1 0 0g体重·天 (N =7)。以上结果表明中缅树鼠句在冷胁迫影响下 ,以增加能量摄入、能量储存和维持能和降低排泄能量的生理机制来维持能量代谢平衡 ,以此对策来提高低温环境适应能力     

树实验种群的繁育

探讨了树实验种群的繁育。结果是5对成年树在7个月内共产11窝31只仔树。27只出生后即人工喂养的仔树中,25只健康存活至进入动物实验。成年树平均产仔周期为（52±8.9）d;仔树的人工喂养成活率为92.6%。可以认为树实验种群的人工繁育是可行的。人工繁育的树有望取代野外捕获的树用于有关的动物实验研究。     

雌性树的麻醉和生殖器官解剖与卵母细胞形态特征的观察

目的　对雌性树的麻醉、生殖器官解剖与卵母细胞形态特征进行观察 ,为制备转基因树奠定基础。方法　用 1%的戊巴比妥钠 (10 - 3ml g体重 )肌内注射对树麻醉 ,比较在不同环境温度下对麻醉效果的影响。对雌性树生殖器官解剖结构、卵母细胞形态特征等进行观察。结果　①在 2 5～ 2 8℃、18～ 2 0℃下麻醉的持续时间分别为 80min、130min ;②雌性树的子宫为双角子宫 ,卵巢外有包膜 ;③卵母细胞富含色素 ,卵母细胞的透明带与质膜的韧性较山羊强。结论　在 2 5～ 2 8℃下 ,用 1%的戊巴比妥钠 (10 - 3ml g体重 )对树麻醉时间比较适中 ,便于实验操作而且苏醒快。雌性树生殖器官解剖结构、卵母细胞形态特征观察表明 ,树的胚胎移植、转基因等实验操作与小鼠类似 ,但树的受精卵须进行离心处理。     

A long‐term record of Quercus decline,logging and fires in a southern Swedish Fagus‐Picea forest

Abstract. We reconstructed forest development and disturbance events (fire and logging) during the last 1000 yr with tree‐ring data, pollen and charcoal analysis from a semi‐natural Fagus sylvatica‐Picea abies forest (ca. 1 km²) in the hemiboreal zone. According to pollen analysis, Quercus robur together with Pinus sylvestris was abundant in the forest until the turn of the 18th/19th centuries when these species disappeared completely (Quercus) or nearly completely (Pinus) and were replaced by Fagus and Picea. The disappearance of Quercus was corroborated by the remarkable discovery of a single Quercus stump that had been cut in the 18th century and had become overgrown and preserved by a very old Picea. In total 11 fires were dated from 1555 to 1748 from fire scars in several Pinus stumps cut 100 ‐ 200 yr ago. Since the last fire in 1748, no Quercus or Pinus have regenerated in the core of the reserve apart from single pines in neighbouring managed forest (80 yr ago). During the period of documented fires Fagus was protected from fires in a refuge made up of large boulders. Picea colonized the region at the time when the fires ceased 250 yr ago. We hypothesize that most of the fires were probably of human origin because of their patchiness and high frequency compared to the natural background levels of lightning ignitions in the region. On a 300‐yr time scale, logging and fire suppression seem to strongly overshadow the effect of climate change on forest composition and dynamics.     

Equilibrium sorption isotherm for metal ions on tree fern

A new sorbent system for removing heavy metal ions, such as Zn(II), Cu(II) and Pb(II), from aqueous solutions has been investigated. This new sorbent is tree fern, an agriculture product. Variables of the system include solution temperature and sorbent particle size. The experimental results were fitted to the Langmuir, Freundlich and Redlich–Peterson isotherms to obtain the characteristic parameters of each model. Both the Langmuir and Redlich–Peterson isotherms were found to well represent the measured sorption data. According to the evaluation using the Langmuir equation, the maximum sorption capacities of metal ions onto tree fern were 7.58 mg/g for Zn(II), 10.6 mg/g for Cu(II) and 39.8 mg/g for Pb(II). It was noted that an increase in temperature resulted in a higher metal loading per unit weight of the sorbent. Decreasing the particle sizes of tree fern led in an increase in the metal uptake per unit weight of the sorbent.