北京西山绿化树种秋季滞纳PM2.5能力及其与叶表面AFM特征的关系

以北京西山6种绿化树种白皮松、油松、柳树、五角枫、银杏、山杨为对象,应用气溶胶再发生器对植物叶片秋季PM_2.5吸附量进行定量研究,同时应用原子力显微镜(AFM)观察叶表面微形态特征,分析了叶表面粗糙度等参数,阐释了各树种叶片吸附PM_2.5的机制.结果表明: 不同树种单位叶面积PM_2.5吸附量排序为白皮松(2.44±0.22 μg·cm^-2)＞油松(2.40±0.23 μg·cm^-2)＞柳树(1.62±0.09 μg·cm^-2)＞五角枫(1.23±0.01 μg·cm^-2)＞银杏(1.00±0.07 μg·cm^-2)＞山杨(0.97±0.03 μg·cm^-2);从秋季不同月份来看,不同树种单位叶面积PM_2.5吸附量表现为11月(2.33±0.43 μg·cm^-2)＞10月(1.62±0.64 μg·cm^-2)＞9月(1.51±0.50 μg·cm^-2).白皮松和油松有大量凹陷和突起,相对高差较大,粗糙度较大,吸滞PM_2.5能力强;柳树和五角枫叶片有褶皱,粗糙度相对较高,分布有大量的突起和凹陷,吸滞PM_2.5能力居中;银杏和山杨因其叶表面平滑、气孔多为长圆形,粗糙度较小,吸滞PM_2.5能力较弱.不同树种正背面粗糙度平均值为白皮松(149.91±16.38 nm)＞油松(124.47±10.52 nm)＞柳树(98.85±5.36 nm)＞五角枫(93.74±21.75 nm)＞银杏(80.84±0.88 nm)＞山杨(67.72±8.66 nm),这与不同树种单位叶面积PM_2.5吸附量排序完全一致,叶片粗糙度与单位叶面积PM_2.5吸附量呈显著正相关(R²=0.9498).为提高城市植被的环境效应,可选择叶表面形态有利于吸滞PM_2.5等颗粒物的树种.     

北京西山绿化树种秋季滞纳PM2.5能力及其与叶表面AFM特征的关系

以北京西山6种绿化树种白皮松、油松、柳树、五角枫、银杏、山杨为对象,应用气溶胶再发生器对植物叶片秋季PM_2.5吸附量进行定量研究,同时应用原子力显微镜(AFM)观察叶表面微形态特征,分析了叶表面粗糙度等参数,阐释了各树种叶片吸附PM_2.5的机制.结果表明: 不同树种单位叶面积PM_2.5吸附量排序为白皮松(2.44±0.22 μg·cm^-2)＞油松(2.40±0.23 μg·cm^-2)＞柳树(1.62±0.09 μg·cm^-2)＞五角枫(1.23±0.01 μg·cm^-2)＞银杏(1.00±0.07 μg·cm^-2)＞山杨(0.97±0.03 μg·cm^-2);从秋季不同月份来看,不同树种单位叶面积PM_2.5吸附量表现为11月(2.33±0.43 μg·cm^-2)＞10月(1.62±0.64 μg·cm^-2)＞9月(1.51±0.50 μg·cm^-2).白皮松和油松有大量凹陷和突起,相对高差较大,粗糙度较大,吸滞PM_2.5能力强;柳树和五角枫叶片有褶皱,粗糙度相对较高,分布有大量的突起和凹陷,吸滞PM_2.5能力居中;银杏和山杨因其叶表面平滑、气孔多为长圆形,粗糙度较小,吸滞PM_2.5能力较弱.不同树种正背面粗糙度平均值为白皮松(149.91±16.38 nm)＞油松(124.47±10.52 nm)＞柳树(98.85±5.36 nm)＞五角枫(93.74±21.75 nm)＞银杏(80.84±0.88 nm)＞山杨(67.72±8.66 nm),这与不同树种单位叶面积PM_2.5吸附量排序完全一致,叶片粗糙度与单位叶面积PM_2.5吸附量呈显著正相关(R²=0.9498).为提高城市植被的环境效应,可选择叶表面形态有利于吸滞PM_2.5等颗粒物的树种.     

北京西山不同海拔油松林PM2.5浓度及叶片吸附量变化规律

以北京西山不同海拔梯度油松人工林为研究对象,对油松林PM_(2.5)浓度变化和叶片PM_(2.5)吸附量进行分析,并应用电子显微镜对不同海拔油松叶表面微形态特征进行观察,阐释叶片吸附PM_(2.5)差异。结果表明:随着海拔升高PM_(2.5)质量浓度逐渐降低,不同海拔油松林PM_(2.5)质量浓度日变化均呈典型的双峰曲线,7:00和19:00是一天的两个峰值,最小值出现在13:00—15:00左右;从不同月份看,不同海拔油松林PM_(2.5)质量浓度最高值出现在冬季的2月,最低值在8月;不同海拔油松林PM_(2.5)质量浓度全年均值为84 m((102.28±18.44)μg/m~3)110 m((94.18±18.34)μg/m~3)160 m((81.53±19.23)μg/m~3)230 m((75.39±15.71)μg/m~3);随着海拔升高单位叶面积PM_(2.5)吸附量逐渐减小,每升高50 m,单位叶面积PM_(2.5)吸附量降低23.25%,每公顷PM_(2.5)吸附量下降26.43%,不同海拔油松林每公顷PM_(2.5)吸附量全年均值为84 m((8.61±1.08)kg/hm~2)110 m((7.30±0.94)kg/hm~2)160 m((6.35±0.99)kg/hm~2)230 m((4.34±1.14)kg/hm~2);处于低海拔的油松叶表面较粗糙,气孔内部和周围聚集大量颗粒物,在叶面形态上更有利于吸附PM_(2.5),高海拔则相反。高海拔空气质量优于低海拔,低海拔的植物吸附颗粒物多于高海拔。研究结果可为城市造林和森林净化大气提供数据支持。     

不同径级白皮松滞留空气中颗粒物的特征

城市园林植物是去除大气PM_(2.5)等颗粒物的有效途径之一,但目前的研究主要集中于以单叶为主的微观尺度上,对于绿化树种选择和环境效益评价具有重要意义的单株尺度研究不足。本文以北京市3个不同污染环境中的白皮松为研究对象,通过叶面尘收集、叶微结构电镜观察、叶面积测定等方法,研究了不同胸径下的白皮松在单叶尺度和单株尺度上的滞尘特征。结果表明:白皮松单位叶面积的滞纳PM_(2.5)、PM_(10)和TSP的最大数量分别为0.15、0.29和0.97 g·m~(-2);胸径对单位叶面积滞留PM_(2.5)等颗粒物的影响不显著(P0.05);不同胸径白皮松叶表面的气孔大小较一致,气孔分布密度相近,且沟状突起分布均匀,没有显著差异;胸径对植物单株滞留量影响较大(P0.05),3个研究点的白皮松滞纳颗粒物水平较高的胸径多集中于12.7~25.5 cm;且单株植物对于PM_(2.5)、PM_(10)和TSP的最大滞尘量分别为5.77、12.88和43.08 g。白皮松的胸径对其叶的微结构影响较小,但显著影响冠幅半径、叶面积指数,使得胸径对单叶尺度上单位叶面积滞尘量的影响不显著,而在单株尺度上具有显著影响。     

亚热带常绿树种对不同粒径颗粒物的滞留能力

可吸入颗粒物和细颗粒物是大部分城市的首要污染物,对人体健康和环境都有重要影响;而城市植物能吸附大气颗粒物,进而有效降低大气颗粒物浓度。为了深入探究不同树种叶表面特征与自身滞尘效益之间的关系,该研究以浙江省三种常见城市绿化树种(青冈、冬青、红花檵木)为对象,采用重量法提取各样本在3个粒径上(8~100,2.5~8,0.45~2.5μm)的单位叶面积滞尘量(μg·cm~(-2)),并结合叶面积指数估测全株滞尘量。结果表明:三种供试植物叶片对颗粒物平均单位叶面积滞留量在30.4~63.7μg·cm~(-2)之间,而平均单木滞尘量每株在1.36-9.36 g之间。红花檵木因其叶表粗糙、具有绒毛等特征,对颗粒物(0.45~100μm)有最大的吸附能力(63.74±12.0μg·cm~(-2));对于大颗粒物(8~100μm)和细颗粒物(0.45~2.5μm),三种植物叶片均对其分别具有最大(40.9%~57.5%)、最小(15.6%~20.6%)的吸附能力;对于单木滞尘量,青冈因其具有较大叶面积指数等特征,对颗粒物总吸附效果更佳(每株9.36g)。该研究结果表明城市绿化树种对减缓大气颗粒物污染起到重要作用。     

北京市常见绿化树种叶片秋季滞纳不同粒径颗粒物能力

选择不同区域典型园林区,于秋季应用气溶胶再发生器(QRJZFSQ-I)测定北京市常绿化树种叶片单位面积滞纳不同粒径颗粒物能力,并推算单位面积林地滞纳颗粒物的物质量。结果表明:针叶树种对各粒径颗粒物(PM_(10)、PM_(2.5)和PM_(1.0))的滞纳能力均高于阔叶树;针叶树种间对不同粒径颗粒物滞纳量没有显著差异,不同阔叶树在南海子公园(城区)差异显著,其中,对PM_(10)滞纳力强的树种为杨树(1.256 kg·hm~(-2)·a~(-1))和柳树(1.153 kg·hm~(-2)·a~(-1)),对PM_(2.5)和PM_(1.0)滞纳力强的树种为白蜡(0.367和0.107 kg·hm~(-2)·a~(-1));相同树种在不同地点对总悬浮颗粒物(TSP)和PM_(10)的滞纳规律相同,对PM_(2.5)和PM_(1.0)的滞纳规律与前两种类型颗粒物略有不同,桧柏对这两种颗粒物的最大值出现在南海子公园(11.043 kg·hm~(-2)·a~(-1)),是最小值北京植物园的15倍;树种滞纳的各种颗粒物以PM_(10)比例较大,且其比例变化按"城区-近郊-远郊"呈渐降的趋势。     

武汉市15种阔叶乔木滞尘能力与叶表微形态特征

以武汉市15种常见的阔叶乔木为研究对象,通过3级滤膜过滤法测定了各乔木单位叶面积滞留不同粒径颗粒物(TSP、PM_(10)、PM_(10)、PM_(2.5))的质量,并通过扫描电镜观察比较了15种乔木的叶表面微形态结构,分析了微形态对植物滞尘能力的影响。结果表明:15种乔木单位叶面积的滞尘量存在显著差异(P0.05),综合滞尘能力最强的植物为二球悬铃木、桂花和石楠,除以上3者外,女贞和广玉兰分别具有较强的滞留PM_(10)和PM_(2.5)的能力;加杨滞留TSP和PM_(10)的能力最弱,玉兰滞留PM_(10)和PM_(2.5)的能力最弱。各乔木单位叶面积滞留PM_(2.5)和PM_(10)的质量分别占总粉尘量的0.7%—8.9%和3.6%—33.9%。叶表面微结构观察表明,叶表面粗糙、褶皱较多,或被有蜡质层的植物有利于粉尘颗粒物的附着。相关性分析表明,植物单位叶面积的滞尘量与叶表面沟槽的宽度呈显著相关,上下表面沟槽宽度越小,越有利于细微颗粒物(PM_(2.5))的滞留,下表面沟槽宽度增加,有利于粉尘总颗粒物(TSP)的滞留。由此可见,叶表面粗糙度、蜡质含量和沟槽宽度等微形态结构是调控绿化树种叶片滞尘能力的重要因素,在武汉以治理大气粉尘污染为目标进行城市绿化时,可考虑选择二球悬铃木、桂花和石楠等滞尘能力强的树种。     

长沙市不同污染程度区域桂花和香樟叶表面PM2.5吸附量及其影响因素

为阐释不同污染程度下城市绿化植物吸滞PM_2.5机理、解析污染物来源,应用气溶胶再发生器定量测定长沙市常见的2种园林绿化树种(桂花和香樟)植物叶片PM_2.5吸附量,同时应用原子力显微镜(AFM)观察了不同污染区(交通区、文教区、清洁区)植被的叶表面微形态特征,使用离子色谱仪测定样品中水溶性离子含量.结果表明: 污染程度与植物叶表面PM_2.5吸附量呈正相关,不同植物单位叶面积PM_2.5吸附量全年均值表现为交通区(0.56±0.04 μg·cm^-2)＞文教区(0.48±0.06 μg·cm^-2)＞清洁区(0.33±0.02 μg·cm^-2),植物单位叶面积PM_2.5吸附量季节变化为冬季(0.70±0.10 μg·cm^-2)＞春季(0.43±0.14 μg·cm^-2)＞秋季(0.39±0.12 μg·cm^-2)＞夏季(0.31±0.09 μg·cm^-2),桂花的单位叶面积PM_2.5吸附量大于香樟;污染程度轻的区域的植物叶片比较光滑,污染程度重的区域的叶片较粗糙,植物粗糙度排序为交通区(195.45±16.09 nm)＞文教区(176.99±8.45 nm)＞清洁区(131.88±12.98 nm);不同污染程度地区PM_2.5离子含量均表现为冬季最大,其次是春季和秋季,夏季最低;3个污染区PM_2.5离子成分均以Na⁺、NH₄⁺、Cl^-和Br^-这4种离子为主,不同程度污染区PM_2.5污染均以移动源污染为主.     

长沙市不同污染程度区域桂花和香樟叶表面PM2.5吸附量及其影响因素

为阐释不同污染程度下城市绿化植物吸滞PM_2.5机理、解析污染物来源,应用气溶胶再发生器定量测定长沙市常见的2种园林绿化树种(桂花和香樟)植物叶片PM_2.5吸附量,同时应用原子力显微镜(AFM)观察了不同污染区(交通区、文教区、清洁区)植被的叶表面微形态特征,使用离子色谱仪测定样品中水溶性离子含量.结果表明: 污染程度与植物叶表面PM_2.5吸附量呈正相关,不同植物单位叶面积PM_2.5吸附量全年均值表现为交通区(0.56±0.04 μg·cm^-2)＞文教区(0.48±0.06 μg·cm^-2)＞清洁区(0.33±0.02 μg·cm^-2),植物单位叶面积PM_2.5吸附量季节变化为冬季(0.70±0.10 μg·cm^-2)＞春季(0.43±0.14 μg·cm^-2)＞秋季(0.39±0.12 μg·cm^-2)＞夏季(0.31±0.09 μg·cm^-2),桂花的单位叶面积PM_2.5吸附量大于香樟;污染程度轻的区域的植物叶片比较光滑,污染程度重的区域的叶片较粗糙,植物粗糙度排序为交通区(195.45±16.09 nm)＞文教区(176.99±8.45 nm)＞清洁区(131.88±12.98 nm);不同污染程度地区PM_2.5离子含量均表现为冬季最大,其次是春季和秋季,夏季最低;3个污染区PM_2.5离子成分均以Na⁺、NH₄⁺、Cl^-和Br^-这4种离子为主,不同程度污染区PM_2.5污染均以移动源污染为主.     

北京常见阔叶绿化植物滞留PM2.5能力与叶面微结构的关系

叶片是植物滞留大气颗粒物的主要载体,对城市环境质量的改善发挥着巨大作用。该文用洗脱法测定了北京市20种常见阔叶绿化植物单位叶面积滞留总悬浮颗粒物(TSP)及PM2.5(颗粒直径≤2.5μm)的质量,并利用扫描电子显微镜观察了叶表面的微结构,分析比较了20种道路绿化植物叶片去除TSP与PM2.5的能力,以探讨典型植物叶表面微结构特征对大气颗粒物拦截效果的影响机理。结果显示:(1)不同植物单位叶面积滞留TSP和PM2.5的量均存在显著差异,变化范围分别为0.40~3.44g/m2和0.04~0.39g/m2。(2)叶表面沟槽宽度的不同可能是不同植物滞留TSP和PM2.5差异的主要原因,沟槽宽度过宽和过窄均不利于叶片捕集颗粒物,且颗粒物滞留量随沟槽深度增加而增大。(3)气孔密度较大的叶片表面颗粒物滞留量较大。研究表明,灌木与藤本植物单位叶面积对TSP和PM2.5的平均滞留量均大于乔木;叶表面沟槽宽度为5μm左右时对PM2.5滞留量较大;悬铃木(Platanus acerifolia)、木槿(Hibiscus syriacus)和大叶黄杨(Buxus sinica)单位叶面积滞留TSP与PM2.5量与其他供试植物相比均较大。     

Laser microirradiation of kinetochores in mitotic PtK2 cells

Irradiation of the kinetochore region of PtK₂ chromosomes by laser light of 532 nm was used to study the function of the kinetochore region in chromosome movement and to create artificial micronuclei in cells. When the sister kinetochores of a chromosome were irradiated at prometaphase, the affected chromosome detached from the spindle and exhibited no further directed movements for the duration of mitosis. The chromatids of the chromosome remained attached to one another until anaphase, at which point they separated. No poleward movement of the chromatids was observed, and at telophase they passively moved to one of the daughter cells and were enclosed in a micronucleus. The daughter cell containing the micronucleus was then isolated by micromanipulation and followed through subsequent mitoses. At the next mitosis, two chromosomes, each with two chromatids, condensed in the micronucleus. These chromosomes did not attach to the spindle and showed chromatid separation, but no poleward movements at anaphase. They were again enclosed in micronuclei at telophase. The third generation mitosis was similar to the second. Occasionally, both the irradiation-produced and naturally occurring micronuclei exhibited no chromosome condensation at mitosis. Feulgenstained monolayers of PtK₂ cells with naturally occurring micronuclei showed that some micronuclei stain positive for DNA and others do not. This finding raises questions about the fate of chromosomes in a micronucleus.     

Enhanced Synthesis of the Exopolysaccharide Ethapolan by Acinetobacter sp. 12S Grown on a Mixture of Substrates

Enhanced synthesis of the exopolysaccharide ethapolan by Acinetobacter sp. 12S was observed when the bacterium was grown on a mixture of two energetically nonequivalent substrates (ethanol and glucose) taken in a molar proportion of 3.1 : 1. The efficiency of carbon transformation into EPSs was maximum when sodium ions were absent in the medium, the concentration of nitrogen source was reduced to 0.3–0.45 g/l, and the inoculum was grown on ethanol. Such conditions provided an increase in the maximum specific growth rate and its attainment in earlier cultivation terms. Molasses as a substitution for glucose was inefficient. The activities of the key enzymes of C₂ metabolism in Acinetobacter sp. 12S cells grown on the substrate mixture were 1.1 to 1.7 times lower than they were during growth on ethanol alone. The activity of isocitrate lyase in cells grown on the substrate mixture declined to an even greater extent (by 4–7 times), indicating that the role of the glyoxylate cycle in such cells is insignificant.     

Localization of cholecystokinin receptor subtypes in the endocine pancreas.

This study was undertaken to clarify the controversy in the literature about pancreatic localization of the cholecystokinin (CCK) CCK(A) and CCK(B) receptors. With antibodies used by other investigators, we first established their specificity by Western blotting, indirect immunofluorescence, and confocal microscopy with each antibody's peptide antigen. Co-localization assays between the CCK receptors and the pancreatic hormones insulin, glucagon, and somatostatin revealed that the CCK(A) RAbs 1122 and R1-2 recognized insulin and glucagon cells in rat, pig, and human pancreas but not in the somatostatin cells. Conversely, the three CCK(B) RAbs tested, 9262, 9491, and GR4, identified the somatostatin cells. Abs 9491 and GR4 occasionally co-localized with glucagon, a feature that never occurred with Ab 9262. Finally, the specificity of Ab 9262 for the pancreatic CCK(B) R was confirmed in six different species. It co-localized with somatostatin but never with glucagon in these species. Our data suggest the use of Abs 1122 and 9262 to specifically identify and localize pancreatic CCK(A) and CCK(B) receptors, respectively. Confusion in the literature may result from the lack of specificity of most antibodies used, as established in this study.     

Distribution and air-water exchange of carbon dioxide in the Scheldt plume off the Belgian coast

In the present paper we report partial pressureof CO₂ (pCO₂) data obtained off theBelgian coast during 24 cruises. The temporaland spatial resolution of this data set allowsus to discuss satisfactorily seasonal andinter-annual variability of pCO₂ in thestudy area. The dynamics of pCO₂ aredescribed using two approaches: fixed referencestations and area survey cruises. The air-waterfluxes of CO₂ in the Scheldt estuarineplume and in the outer-plume region areestimated quantitatively, showing that theseareas correspond respectively to a net annualsource and sink of atmospheric CO₂. Theannually integrated air-water fluxes for theScheldt estuarine plume range between +1.1 and+1.9 mol m^–2 year^–1 as a function ofthe formulation of the exchange coefficient ofCO₂. The annual net emission of CO₂from the estuarine plume to the atmosphere isestimated to be between +2.3 to +4.0 Gmolyear^–1 which represents 17 to 29% of theestimate reported in the literature for the Scheldtinner estuary.     

Substrate kinetics of the Acanthamoeba castellanii alternative oxidase and the effects of GMP

In Acanthamoeba castellanii mitochondria, the apparent affinity values of alternative oxidase for oxygen were much lower than those for cytochrome c oxidase. For unstimulated alternative oxidase, the K_Mox values were around 4-5 μM both in mitochondria oxidizing 1 mM external NADH or 10 mM succinate. For alternative oxidase fully stimulated by 1 mM GMP, the KK_Mox values were markedly different when compared to those in the absence of GMP and they varied when different respiratory substrates were oxidized (K_Mox was around 1.2 μM for succinate and around 11 μM for NADH). Thus, with succinate as a reducing substrate, the activation of alternative oxidase (with GMP) resulted in the oxidation of the ubiquinone pool, and a corresponding decrease in K_Mox. However, when external NADH was oxidized, the ubiquinone pool was further reduced (albeit slightly) with alternative oxidase activation, and the K_Mox increased dramatically. Thus, the apparent affinity of alternative oxidase for oxygen decreased when the ubiquinone reduction level increased either by changing the activator or the respiratory substrate availability.     

Spatial variability of soil nitrate nitrogen after potatoes and its change during winter

Knowledge of the frequency distribution and spatial structure of the soil NO₃-N is required to develop an efficient sampling strategy. A 1 ha polder field was sampled after the harvest of potatoes in October 1987, and in February and April 1988, without being fertilitzed since March 1987. These data sets were examined by a classical statistical as well as a spatial structure analysis. The October and February data sets were found to be lognormally distributed, the April data showed a normal frequency distribution. All three data sets had a spatial structure, although the October data were anisotropic and needed removal of a trend. The spatial variability of soil NO₃−N decreased, became isotropic and evolved towards a larger range of spatial dependence during the winter. Knowledge of this structure permitted to krige or cokrige the data. The number of samples required to estimate the mean NO₃−N content with an acceptable precision was found to be 39, 43 and 17 in October, February and April respectively.     

Tryptophan modification of phospholipase A2 enzymes and presynaptic neurotoxins from snake venoms

Three phospholipases A₂ purified from cobra venoms and two presynaptically acting neurotoxins that exhibit phospholipase A₂ activity were subjected to tryptophan modification with 2-hydroxy-5-nitrobenzyl bromide. Associated with the modification of an increasing number of Trp residues were marked decreases in enzymatic activity and lethality, whereas antigenicity remained unchanged. The degree of exposure of tryptophanyl groups as determined by acrylamide quenching was consistent with the relative reactivity toward 2-hydroxy-5-nitrobenzyl bromide, except for Hemachatushaemachatus phospholipase A₂, which showed unusually high reactivity due to its characteristic dimeric conformation. Difference spectra of Trp-modified derivatives differed from those of their native enzymes by the presence of a new positive perturbation between 350 and 500 nm, with a maximum at 415 nm. Scatchard plots revealed only one type of binding site for Ca²⁺, and the binding abilities of the modified enzymes were not impaired. At pH 8.0, all native enzymes enhanced the emission intensity of 8-anilinonaphthalene sulfonate (ANS) dramatically, and the emission intensity of the ANS-enzyme complex increased or decreased in parallel with increasing concentration of Ca²⁺ for the respective enzyme. The Trp-modified derivatives did not enhance the emission intensity of ANS at all either in the presence or absence of Ca²⁺. By means of tryptophan modification, we were able to infer that the tryptophan residues are in the vicinity of the Ca²⁺ binding site and are directly involved in the binding with ANS. This, together with the suggestion that the hydrophobic pocket that interacts with ANS might be the site of binding of the phospholipase A₂ enzyme with the substrate, suggests that the Trp residues in phospholipase A₂ enzymes and presynaptic toxins are involved in substrate binding.     

The stator complex of the A1A0-ATP synthase--structural characterization of the E and H subunits

Archaeal ATP synthase (A-ATPase) is the functional homolog to the ATP synthase found in bacteria, mitochondria and chloroplasts, but the enzyme is structurally more related to the proton-pumping vacuolar ATPase found in the endomembrane system of eukaryotes. We have cloned, overexpressed and characterized the stator-forming subunits E and H of the A-ATPase from the thermoacidophilic Archaeon, Thermoplasma acidophilum. Size exclusion chromatography, CD, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and NMR spectroscopic experiments indicate that both polypeptides have a tendency to form dimers and higher oligomers in solution. However, when expressed together or reconstituted, the two individual polypeptides interact with high affinity to form a stable heterodimer. Analyses by gel filtration chromatography and analytical ultracentrifugation show the heterodimer to have an elongated shape, and the preparation to be monodisperse. Thermal denaturation analyses by CD and differential scanning calorimetry revealed the more cooperative unfolding transitions of the heterodimer in comparison to those of the individual polypeptides. The data are consistent with the EH heterodimer forming the peripheral stalk(s) in the A-ATPase in a fashion analogous to that of the related vacuolar ATPase.     

Effect of temperature and photoperiod on efficiency of assimilated CO<Subscript>2</Subscript> conversion into the biomass of <Emphasis Type="Italic">Cucumis sativus</Emphasis>

The coefficient of effectiveness (K _e) of assimilated CO₂ conversion into dry matter of cucumber (Cucumis sativus L.) plants at the stage of four leaves as dependent on a photoperiod (8, 12, and 16 h) at an irradiance of 220 W/m² at the upper leaf level and the combinations of day and night temperatures: typical temperature of plant habitat (background temperature) of 25°C and heat- and cold-hardening temperatures (35 and 15°C, respectively) was determined in the multifactorial designed experiment. K _e reduced insignificantly at shortening of a photoperiod and greater at its lengthening. At background temperatures, K _e corresponded mainly to that of carbohydrate synthesis while the presence of cold-hardening temperatures in the thermoperiod increased K _e and heat-hardening temperature reduced it.Translated from Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 203–208.Original Russian Text Copyright © 2005 by Talanov, Popov, Kurets, Drozdov.This revised version was published online in April 2005 with a corrected cover date.     

Studies on the status of tyrosyl residues in phospholipases A2 fromNaja naja atra andNaja nigricollis snake venoms

Two phospholipases A₂ (PLA₂) fromNaja naja atra andNaja nigricollis snake venoms were subjected to tyrosine modification withp-nitrobenzenesulfonyl fluoride (NBSF) atpH 8.0. Three major NBS derivatives from each PLA₂ were separated by high-performance liquid chromatography. The results of amino acid analysis showed that only two Tyr residues out of nine were modified, and the modified residues were identified to be Tyr-3 and Tyr-63 (or Tyr-62) in the sequence. Spectrophotometric titration indicated that the phenolic group of Tyr-3 and Tyr-63 (or Tyr-62) had apK of 10.1 and 11.0, respectively. The reactivity of Tyr-3 toward NBSF was not affected in the presence or absence of Ca ²⁺; however, the reactivity of Tyr-63 (or Tyr-62) toward NBSF was greatly enhanced by Ca²⁺. Modification of Tyr-63 (or Tyr-62) resulted in a marked decrease in both lethality and enzymatic activity. Conversely, modification of Tyr-3 inN. naja atra PLA₂ could cause more than a sixfold increase in lethal potency, in sharp contrast to the loss of enzymatic activity.Tyrosine-63-modifiedN. naja atra PLA₂ exhibited the same Ca²⁺-induced difference spectra as that of native PLA₂, indicating that the Ca²⁺-binding ability of Tyr-63-modifiedN. naja atra PLA₂ was not impaired. However, Tyr-3-modified PLA₂ and all Tyr-modifiedN. nigricollis CMS-9 were not perturbed by Ca²⁺, revealing that the Ca²⁺-binding ability have been lost after tyrosine modification. These results suggest that Tyr-62 inN. nigricollis CMS-9 and Tyr-3 in both enzymes are involved in Ca²⁺ binding. AtpH 8.0, both native PLA₂ enzymes enhance the emission intensity of 8-anilinonaphthalene sulfonate (ANS) dramatically, while all of the Tyr-modified derivatives did not enhance the emission intensity at all either in the presence or absence of Ca²⁺, suggesting that the hydrophobic pocket that interacts with ANS might be the substrate binding site, in which Tyr-3 and Tyr-63 (or Tyr-62) are involved.