Codon usage in the vertebrate hemoglobins and its implications

A study of codon usage in vertebrate hemoglobins revealed an evolutionary trend toward elevated numbers of CpG codon boundary pairs in mammalian hemoglobin alpha genes. Selection for CpG codon boundaries countering the generally observed CpG suppression is strongly suggested by these data. These observations parallel recently published experimental results that indicate that constitutive expression of the human alpha-globin gene appears to be determined by regulatory information encoded within the structural gene. The possibility is raised that, in the absence of selection, CpG decay can be used to date the evolutionary origin of a mammalian alpha pseudogene from its active alpha gene.      

Evolution of crystallins: expression of lens-specific proteins in the blind mammals mole (Talpa europaea) and mole rat (Spalax ehrenbergi)

The mole (Talpa europaea; Insectivora) and the mole rat (Spalax ehrenbergi; Rodentia) both have degenerated eyes as a convergent adaptation to subterranean life. The rudimentary eye lenses of these blind mammals no longer function in a visual process. The crystallin genes, which display a lens-specific expression pattern, were studied in these blind mammals and in related species with normal eyes by hybridizing their genomic DNAs with probes obtained from cDNA clones for alpha A-, alpha B-, and beta Bp-crystallins from calf and gamma 3- crystallin from the rat. For all crystallin genes examined, the hybridization signals of mole and mole rat genomic DNA were comparable, respectively, with those of shrew and of rat and mouse, normal-vision representatives of the orders Insectivora and Rodentia. The expression of the crystallins at the protein level was tested by using antiserum specific for alpha-crystallin in immunofluorescence reactions on lens sections of mole and mole rat eyes and by using antisera against the beta- and gamma-crystallins on sections of the mole eye. All antisera gave positive fluorescence reactions exclusively with lens tissue of these blind mammals, indicating that the crystallins are still normally expressed despite the fact that these lenses have had no function in a visual process in these mammals for at least many million years. These findings apparently imply that some unknown selective advantage has conserved the crystallin genes and their expression after the loss of normal function of the lenses.      

The evolutionary history of the amylase multigene family in Drosophila pseudoobscura

In Drosophila pseudoobscura, the amylase (Amy) multigene family is contained within a series of inversions, or gene arrangements, on the third chromosome. The Standard (ST), Santa Cruz (SC), and Tree Line (TL) inversions are central to the phylogeny of arrangements, and have clusters of other arrangements derived from them. The gene arrangements belonging to each of these three clusters have a characteristic number of Amy genes, ranging from three in ST to two in SC to one in TL. This distribution pattern can reflect a history of either duplications or deletions, although the data available in the past did not permit a decision between these alternatives. We provide unambiguous evidence that three Amy genes were present before the divergence of the ST, SC, and TL arrangements. Thus, the current status of the Amy multigene family is the result of deletions in the TL and SC arrangements, which created three new pseudogenes: TL Amy2-psi, TL Amy3-psi, and SC Amy3- psi. Analysis of pseudogene sequences revealed that, in the SC and ST arrangements, pseudogene evolution has been retarded, most likely due to the homogenization effect of gene conversion. Finally, by determining the original copy number, we have reconstructed the evolutionary history of the Amy multigene family and linked it with the evolution of the central gene arrangements.      

Evidence for gene conversion in the amylase multigene family of Drosophila pseudoobscura

The alpha-amylase (Amy) multigene family in Drosophila pseudoobscura is located on the third chromosome, which is polymorphic for more than 40 inverted gene arrangements. The number of copies in this family ranges from one to three, depending on the arrangement in question. A previous study of the three Amy genes from the Standard (ST) arrangement suggested either that duplicated copies (Amy2 and Amy3) are functionally constrained or that they are undergoing gene conversion with Amy1. In order to elucidate further the pattern of molecular evolution in this family, we cloned and sequenced four additional Amy genes, two from the Santa Cruz (SC) and two from the Chiricahua (CH) gene arrangement. Of the two alternatives, only the hypothesis of gene conversion is supported by the sequence analysis. The homogenization effect of gene conversion has been strongest in SC, whose copies differ by only two nucleotides, less noticeable in ST, and negligible in the CH. Furthermore, the action of gene conversion is apparently localized, occurring only in the coding region. Interestingly, these results concur with the findings of other workers for the duplicated Amy genes in the Drosophila melanogaster group. Thus, the occurrence of gene conversion in the Amy multigene family seems to be a common feature in the Drosophila species studied so far.      

Effect of leaf water deficit on stomatal and nonstomatal regulation of net carbon dioxide assimilation

The effect of leaf water deficit on net CO₂ assimilation was studied under two conditions: in one, the stomata were allowed to contribute to the regulation of CO₂ assimilation; in the other, air was forced through the leaf at a constant rate to overcome the effects of change in stomatal resistance accompanying changes in leaf water deficit. When the stomata were allowed to regulate the gaseous diffusive resistance of the leaf, CO₂ assimilation decreased with increasing leaf water deficit. However, when air was forced through the leaf, the rate of assimilation was not inhibited by increasing leaf water deficit. The results indicate that the inhibition of net CO₂ assimilation with increasing leaf water deficit is a consequence of an increase in the diffusive resistance to gas exchange and not of a change in apparent mesophyll resistance.     

Halothiophene benzimidazoles as P1 surrogates of inhibitors of blood coagulation factor Xa

Neutral weak halothiophene benzimidazole inhibitors of the serine protease factor Xa were identified via screening of a compound library. The X-ray crystal structure of representative 3a bound to human fXa confirmed the S1 binding mode. Starting from 3a a series of halothiophene benzimidazoles was synthesized and investigated for their factor Xa inhibitory activity. This led to potent and selective achiral inhibitors against fXa such as compounds 9k and 9w.     

Expression,secretion and surface display of a human alkaline phosphatase by the ciliate <Emphasis Type="Italic">Tetrahymena thermophila</Emphasis>

Background  

Tetrahymena thermophila possesses many attributes that render it an attractive host for the expression of recombinant proteins. Surface proteins from the parasites Ichthyophthirius multifiliis and Plasmodium falciparum and avian influenza virus antigen H5N1 were displayed on the cell membrane of this ciliate. Furthermore, it has been demonstrated that T. thermophila is also able to produce a functional human DNase I. The present study investigates the heterologous expression of the functional human intestinal alkaline phosphatase (hiAP) using T. thermophila and thereby presents a powerful tool for the optimization of the ciliate-based expression system.     

代谢、血液碱化和纯氧影响呼吸调控的人体实验研究II： 血液碱化后运动试验*

目的: 在完成吸入室内空气状态下症状限制性最大极限心肺运动试验(CPET)和动脉血气指标动态变化规律的基础上,进一步探讨体液酸碱度和CO₂含量对呼吸调控的影响。方法: 选正常志愿者5名,给予5%NaHCO₃(总量0.3 g/kg)分次口服,每5 min口服75 ml(3.75g )。总量服完1 h后,重复CPET。于静息、热身、运动及恢复期,连续测定肺通气指标及每分钟动脉取样的血气指标变化,并与本人在非碱化血液条件下对照数据进行配对t检验比较。结果: 碱化血液之后,CPET期间随着运动功率逐步递增,气体交换和血气指标的反应模式与非碱化血液对照相似（P>0.05）;即与静息状态比较,每分通气量、潮气量、呼吸频率、VO₂、VCO₂均呈现近于线性渐进性递增(P<0.05~0.001)。与碱化血液前吸入室内空气的对照比较：在碱化血液条件下,所有时间点血红蛋白浓度,PaCO₂与pH均显著提高(P<0.05);除无氧阈PaCO₂减低外,只有热身状态呈增高态势,统计学有显著差异(P<0.05);而PaO₂无差异(P>0.05),各状态均较对照状态减低,除恢复期外均有统计学差异(P<0.05)。与非碱化血液对照比较,除静息每分通气量低于对照(P<0.05）外,所有通气指标均无统计学差异(P>0.05)。结论: 碱化血液条件下, 尽管有更高的CaCO₂, PaCO₂ 和 pHa平均水平及更低的Hba和[H⁺]a平均水平,机体对CPET的呼吸反应模式基本相似。     

代谢、血液碱化和纯氧影响呼吸调控的人体实验研究III： 血液碱化后纯氧运动试验*

目的: 在急性血液碱化前、后空气吸入下完成症状限制性最大极限心肺运动试验(CPET)的基础上,本文探讨在血液碱化后吸入纯氧对呼吸调控的影响。方法: 正常志愿者5名在碱化血液后呼吸纯氧CPET,在静息、热身、运动及恢复期,连续测定肺通换气指标及每分钟动脉取样的血气指标,对CPET期间的呼吸气体交换和血气指标的动态变化进行分析,同时与急性碱化血液前、后空气CPET数据比较。结果: 碱化血液后吸入纯氧运动呼吸反应与急性碱化血液前、后空气CPET呼吸反应基本一致。CPET期间,各运动状态下的每分通气量均与对照组相似(P>0.05);仅静息每分通气量较血液碱化空气CPET略高(P<0.05),而其它状态和恢复2min时均相近(P>0.05)。潮气量仅峰值运动时较对照和血液碱化空气CPET略低(P<0.05);而运动过程和恢复2min时的潮气量均相近(P>0.05)。呼吸频率在各个时间与血液碱化前后CPET均无差异(P>0.05)。在碱化血液后吸入纯氧运动各个时期的PaO₂和SaO₂较碱化血液前后空气CPET时明显提高(P<0.001,P<0.05)。血红蛋白浓度虽然较急性血液碱化前后均低,但仅较血液碱化前显著降低(P<0.05),比血液碱化后差异不显著(P>0.05) ; 开始时的PaCO₂较碱化血液前后空气CPET时降低(P<0.05),无氧阈时相近(P>0.05),但到峰值及恢复2 min时明显增高(P<0.05);pH仅较对照增高(P<0.05),但与碱化血液空气试验时无差异;乳酸水平较对照略高,但仅在热身和恢复期有差异(P<0.05)。纯氧提高了两人无氧阈和三人峰值运动的功率和时间。结论: 虽然血液碱化给予纯氧, CPET呼吸反应与碱化血液前、后空气CPET呼吸反应模式相似,表明运动中呼吸反应主要取决于代谢变化,而非动脉血气平均值高低。