钉螺对夹竹桃化感物质三萜总皂甙毒理作用的反应

本文分别用0.02 mg/L-0.1mg/L系列浓度的夹竹桃三萜总皂甙水溶液处理钉螺 ,设无氯清水饲养与浓度为1 mg/L 氯硝柳胺溶液浸杀对照,研究夹竹桃三萜总皂甙是否为夹竹桃灭螺的化感物质及其作用机理.结果表明:用不同浓度梯度的夹竹桃三萜总皂甙水溶液处理钉螺均有一定的杀灭作用, 对试验数据进行概率单位回归分析,表明浸杀处理2、3、 4和5 d的LC50分别为78.31、30.26、20.50、14.19 mg/L,其对应的95%置信区间分别为 63.60-108.19、9.49-44.42、2.86-30.90、0.23-22.79 mg/L;以不同处理浓度、时间下钉螺的死亡率为因变量,对夹竹桃三萜总皂甙水溶液浸杀灭螺效果进行方差分析,结果显示夹竹桃三萜总皂甙的浓度、处理时间的长短对钉螺的死亡率均有极显著的作用效果,并且药液浓度与处理时间之间的交互效应也极显著.扫描电镜观察结果显示40 mg/L浓度处理后钉螺软体表层呈现明显的损伤;透射电镜观察40 mg/L浓度处理后钉螺样品发现24 h后肝细胞核肿胀,核仁消失,核质稀疏,内质网断裂,线粒体增多;当处理时间延长到48 h时,损伤加重,内质网几乎全部囊泡化,细胞核开始破裂,线粒体也破裂;处理24 h后钉螺小肠表面的微绒毛排列有些不整齐,有一小部分微绒毛出现脱落,肠周缘细胞出现肿胀,细胞核出现少量小空泡;处理48 h时的钉螺小肠表面微绒毛排列明显不整齐,大部分微绒毛脱落,肠周缘细胞肿胀明显,有大量空泡,有时还出现游离核,胞质内的细胞器有大小不等的空泡样变性 ,细胞器结构已无法辨别,严重变性,胞膜不完整.采用聚丙稀酰胺电泳技术分离经40 mg/ L夹竹桃三萜总皂甙水溶液处理1、2、3、4、5 d后的钉螺样品的酯酶同工酶,处理1-2 d后酶的活性比清水对照增强,应激反应明显,而处理达3 d时酶的活性开始下降,5 d时,酶谱带数及其色泽与用1 mg/L氯硝柳胺水溶液处理1 d的对照相当.本研究获得化感作用植物夹竹桃灭螺的超微结构证据[动物学报 54(3):489-499,2008].     

益母草(Leonurus artemisia) 灭螺效果研究

本实验研究分别采用益母草的根、茎、叶的0.10、0.25、0.50、2.00、1.00g/L和水苏碱的0.20、0.40、0.60、0.80、1.00g/L5个不同浓度梯度的处理液处理钉螺,设清水和0.001g/L浓度的氯硝柳胺溶液为对照。结果表明:(1)益母草各部分的水浸液均有很好的灭螺效果,用不同浓度的处理液浸杀钉螺,在不同时间的处理下,钉螺死亡率存在差异,其钉螺死亡率是随处理浓度的增加和时间的延长呈上升趋势,0.5g/L以上的益母草根、茎、叶、化水浸液和浓度达0.60g/L以上的水苏碱处理液均可达到100%的明显毒杀钉螺致死效果,与通常使用浓度0.0001g/L氯硝柳胺溶液的灭螺效果相当,不过益母草根、茎、叶水浸液的毒效较氯硝柳胺略慢,用0.0001g/L氯硝柳胺溶液处理钉螺2~3d可达100%的死亡率,而用0.5g/L以上的益母草根、茎、叶水浸液水溶液处理需要3~5d才能达到同样的效果;灭螺效果顺序依次为:叶>茎>根。(2)钉螺趋避性研究表明水苏碱和益母草根、茎和叶的处理液对钉螺具有明显的驱逐作用,而盐酸益母草碱几乎没有作用。由此获得化感作用植物益母草灭螺的化学生态学证据,为研制新的具中国特色的植物成份灭螺剂打下了基础,并为合成仿生灭螺剂以及最终构建生态工程中强化感作用植物群落灭螺提供理论依据。     

益母草（Leonurus artemisia）灭螺效果研究

本实验研充分别采用益母草的根、茎、叶的0．10、0．25、0．50、2．00、1．00g／L和水苏碱的0．20、0．40、0．60、0．80、1．00g/L 5个不同浓度梯度的处理液处理钉螺,设清水和0．001g／L浓度的氯硝柳胺溶液为对照。结果表明：（1）益母草各部分的水浸液均有很好的灭螺效果。用不同浓度的处理液浸杀钉螺,在不同时间的处理下,钉螺死亡率存在差异,其钉螺死亡率是随处理浓度的增加和时间的延长呈上升趋势,0．5g／L以上的益母草根、茎、叶、化水浸液和浓度达0．60g／L以上的水苏碱处理液均可达到100％的明显毒杀钉螺致死效果,与通常使用浓度0．0001g／L氯硝柳胺溶液的灭螺效果相当,不过益母苹根、茎、叶水浸液的毒效较氯硝柳胺略慢,用0．0001g／L氯硝柳胺溶液处理钉螺2—3d可达100％的死亡率,而用0．5g／L以上的益母草根、茎、叶水浸液水溶液处理需要3—5d才能达到同样的效果;灭螺效果顺序依次为：叶〉茎〉根。（2）钉螺趋避性研究表明水苏碱和益母草根、茎和叶的处理液对钉螺具有明显的驱逐作用,而盐酸益母草碱几乎没有作用。由此获得化感作用植物益母草灭螺的化学生态学证据。为研制新的具中国特色的植物成份灭螺剂打下了基础。并为合成仿生灭螺剂以及最终构建生态工程中强化感作用植物群落灭螺提供理论依据。     

益母草不同组分的抑螺效果及对钉螺酯酶同工酶的影响

通过水浸液灭螺实验,对益母草的根、茎、叶水浸液及益母草水苏碱水溶液的杀灭钉螺作用进行了初步研究.结果表明:各处理均有较好的灭螺效果,但毒效较氯硝柳胺略慢,益母草各部分灭螺效果的顺序为叶＞茎＞根;采用同工酶电泳技术检测了益母草水浸液处理钉螺时对钉螺酯酶同工酶的影响,处理1～2 d后样品的酶活高于对照组,处理3～4 d后的酶活则大大减弱,在这一过程中,有时出现新的酶带,有时又有酶带消失,酶带的变化主要在正极区,与正常的病理反应完全一致.     

樟树水浸液对钉螺的生态毒理学效应

用樟树(Cinnamomum camphora)新鲜根皮、茎皮、叶为材料,设置1%、0.5%、0.1%、0.05%4个不同浓度梯度的水浸液处理钉螺,以清水和1 mg/L浓度的氯硝柳胺溶液为对照,统计钉螺死亡率,并分析钉螺过氧化氢酶(CAT)和过氧化物酶(POD)活性及采用透射电镜分析亚显微结构损伤情况。结果表明:(1)樟树各部分的水浸液均有很好的灭螺效果。钉螺死亡率随处理浓度的增加和时间的延长呈上升趋势,0.5%—1.0%以上的樟树根皮、茎皮、叶水浸液均可达到100%的毒杀钉螺致死效果,与1 mg/L氯硝柳胺溶液的灭螺效果相当;灭螺效果顺序依次为:根皮茎皮叶。(2)在处理初期,钉螺体内过氧化氢酶和过氧化物酶活性明显增强,但随处理时间延长,其活性急剧降低;过氧化物酶同工酶谱显示,处理24—48 h后钉螺酶活高于对照组,处理72—96 h后酶活最强,而处理120 h酶活则大大减弱。(3)钉螺肝细胞亚显微结构观察表明,随处理时间的延长,线粒体、内质网、细胞核等结构损伤越来越严重。这些结果表明樟树水浸液对钉螺肝脏造成过氧化伤害和细胞器结构损伤,具有很好的杀螺效果,可作为生态工程抑螺林树种。     

夹竹桃皂甙对福寿螺的毒杀效果及其对水稻幼苗的影响

室内条件下研究了夹竹桃皂甙对福寿螺的杀灭效果及其对水稻幼苗的影响。结果表明,福寿螺的致死率随着皂甙浓度的增加和处理时间的延长而显著上升,其中50mg/L 浓度皂甙处理48h的致死率达100%,与0.156g/L氯硝柳胺水溶液的灭螺效果相当。皂甙对福寿螺的杀灭效果与福寿螺的大小有关。皂甙对壳高h＜10 mm的福寿螺的杀螺效果最好, 而对20≤h<30 mm的杀螺效果最差。水稻幼苗的存活率随着皂甙浓度的增加而显著增加,且低浓度皂甙处理24h也具有较高的值。此外,采用水培方法研究了皂甙对水稻幼苗鲜重的影响。高浓度(≥40 mg/L)的皂甙处理7d显著地抑制了水稻幼苗根部和地上部分的鲜重,其中对根部的影响大于地上部分。当处理时间延长到14d时,皂甙对水稻幼苗鲜重的抑制作用逐渐减弱,仅50mg/L 皂甙处理对根部鲜重有显著地抑制。而在氯硝柳胺水溶液处理下,水稻幼苗停止生长。综合结果表明夹竹桃皂甙是一种环境友好且能高效防治福寿螺的灭螺剂。     

试用夹竹桃颗粒剂杀灭钉螺初报

利用某些植物有效成分对钉螺的毒杀作用进行灭螺 ,可有效地减轻化学药物灭螺方法对环境的污染和对人畜的毒害。现有利用植物灭螺的方法 ,一般是采用提取植物有效成分或利用植物浸出液杀灭钉螺[1、3] 。本文研制了一种目前尚未见报导的利用夹竹桃 (Ｎｅｒｉｕｍｉｎｄｉｃｕｍ)粉碎物和淀粉、明胶等配合制成的植物颗粒灭螺剂 ,用于灭螺实验取得了较满意的效果 ,为利用植物灭螺提供了一种新的方法和思路。1　材料与方法1 1　供试材料　夹竹桃叶 ,淀粉 ,明胶 ,钉螺。1 2　材料的处理( 1 )材料的处理和配合　将夹竹桃叶烘干磨粉并用 60目纱网…     

植物颗粒剂对钉螺毒杀效果的研究

将3种药用植物材料分别粉碎后与淀粉、明胶配合制成直径2～3mm的植物颗粒剂用于灭钉螺试验,结果表明,钉螺取食3种颗粒剂后,24h死亡率分别达到92.5％～100％.其毒杀效果明显优于相同植物的其它灭螺方法,对钉螺的致死时间比氯硝柳胺化学灭螺剂(1×10-3g·L-1)提前12～24h.用3种植物颗粒灭螺剂进行野外灭螺实验,钉螺死亡率最高可达61.5％.     

钉螺的基础组分及溴乙酰胺对它的作用

本文对安徽省贵池地区钉螺软体的生化组成进行了分析,并进一步探讨杀螺药溴乙酰胺对钉螺的怍用。溴乙酰胺浸泡钉螺不同时间后,钉螺组织内糖原含量随作用时间不同而异。杀螺药氯乙酰胺与氯硝柳胺同样拉可减少钉螺体内的糖原含量。     

夹竹桃灭钉螺效果初报

1　引　　言在防治血吸虫病的过程中 ,人们一直致力于消灭其唯一中间宿主钉螺 .近年来长江中下游生态环境破坏严重 ,一些地方有螺面积扩大 ,疫情回升 .常用化学药物灭螺法虽有较明显的灭螺效果 ,但也造成了严重的环境污染 .90年代以来生物灭螺开始成为比较活跃的研究领域 ,在筛选灭螺植物 ,提取植物灭螺活性成分和利用植物他感作用灭螺等方面已取得了许多成绩[1～ 4 ,7,8] .目前 ,植物灭螺难以推广的主要问题在于利用新鲜植物材料灭螺常需要较高的浓度 ,而原料来源往往有限 ;提取植物灭螺活性成分制作灭螺剂的成本较高 ,难以广泛应用 .因此 …     

ATP/ADP exchange activity of gastric (H+ + K+)-ATPase

The ATP/ADP exchange is shown to be a partial reaction of the $\text{(}\text{H}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ by the absence of measurable nucleoside diphosphokinase activity and the insensitivity of the reaction to $\text{P}{}^1$, $\text{P}{}^5$ -di(adenosine-5′) pentaphosphate, a myokinase inhibitor. The exchange demonstrates an absolute requirement for Mg²⁺ and is optimal at an ADP/ATP ratio of 2. The high ATP concentration ($\text{K}{}_{0.5}\text{= 116 μ}\text{M}$) required for maximal exchange is interpreted as evidence for the involvement of a low affinity form of nucleotide site. The ATP/ADP exchange is regarded as evidence for an ADP-sensitive form of the phosphoenzyme. In native enzyme, pre-steady state kinetics show that the formation of the phosphoenzyme is partially sensitive to ADP while modification of the enzyme by pretreatment with 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) in the absence of Mg²⁺ results in a steady-state phosphoenzyme population, a component of which is ADP sensitive. The ATP/ADP exchange reaction can be either stimulated or inhibited by the presence of K⁺ as a function of pH and Mg²⁺.     

Characterization of cytochrome P-450SCC-containing liposomes

Purified cytochrome $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ from bovine adrenocortical mitochondria was incorporated into liposomes by the cholate-dilution method utilizing either dialysis or Sephadex gel filtration. Among synthetic phospholipids tested, dioleoylglycerophosphocholine showed the best stability during the incorporation of $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ into liposomes. A maximum amount of heme was incorporated into liposomes at a molar ratio of phospholipid to the cytochrome of approx. 200. When $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was incorporated into the dioleoylglycerophosphocholine liposomes by the cholate-filtration method, the $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}\text{-}\text{containing liposomes}$ showed two major populations on the elution pattern of the Sepharose 4B gel filtration, and were seen at a diameter of 200–600 Å and its aggregated forms. When the cytochrome was incorporated into dioleoylglycerophosphocholine liposomes or cholesterol-free adrenocortical mitochondrial liposomes, $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was less stable than $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ in aqueous solution. Cholesterol or adrenodoxin markedly stabilized the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. Liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ required cholesterol for its optimum reduction with adrenodoxin, adrenodoxin reductase, and NADPH in the presence of CO. About 70% of the total heme in the dioleoylglycerophosphocholine liposomes was reduced by the enzymatic reduction in the presence of cholesterol, indicating that 70% of the total molecules are exposed to the surface of the outer monolayer. In order to see the location of the heme in membrane, the dioleoylglycerophosphocholine-liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was subjected to $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ treatment. This reagent destroyed the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. These results suggest that the heme is located in the proximity of the $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ reacting sites which are exposed to the surface, or located on the vincinity of polar heads of the membrane.     

AutoFACT: AnAutomaticFunctionalAnnotation andClassificationTool

Background  

Assignment of function to new molecular sequence data is an essential step in genomics projects. The usual process involves similarity searches of a given sequence against one or more databases, an arduous process for large datasets.     

Ligand-dependent reactivity of (Na+ + K+)-ATPase with showdomycin

Showdomycin inhibited pig brain ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ with pseudo first-order kinetics. The rate of inhibition by showdomycin was examined in the presence of 16 combinations of four ligands, i.e., Na⁺, K⁺, Mg²⁺ and ATP, and was found to depend on the ligands added. Combinations of ligands were divided into five groups in terms of the magnitude of the rate constant; in the order of decreasing rate constants these were: (1)$\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$, (2) $\text{Mg}{}^{\mathrm{2+}}$, $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}$ and none, (3) $\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{,}\text{Na}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}$ and $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}$, (4) $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}\text{,}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}$ and $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}\text{, (5)}\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$ and ATP. The highest rate was obtained in the presence of Na⁺, Mg²⁺ and ATP. The apparent concentrations of Na⁺, Mg²⁺ and ATP for half-maximum stimulation of inhibition ($\text{K}{}_{0.5}{}^{\mathrm{<mtext>s</mtext>}}$) were 3 mM, 0.13 mM and 4μM, respectively. The rate was unchanged upon further increase in Na⁺ concentration from 140 to 1000 mM. The rates of inhibition could be explained on the basis of the enzyme forms present, including E₁, E₂, ES, E₁-P and E₂-P, i.e., E₂ has higher reactivity with showdomycin than E₁, while E₂-P has almost the same reactivity as E₁-P. We conclude that the reaction of ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ proceeds via at least four kinds of enzyme form (E₁, E₂, E₁ · nucleotide and EP), which all have different conformations.     

Characterization of partially purified (Na+ + K+)-ATPase from porcine lens

The partial purification of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from pig lens has been achieved by treatment with deoxycholate followed by density gradient centrifugation. The specific activity of the final preparation, ranging from 300 to 500 nmol/h per mg protein, is increased approx. 100-fold compared to the homogenate. A parallel increase in $\text{p-}\text{nitrophenylphosphatase}$ activity is also observed. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals six major protein bands, one of which is the 93 kDa α subunit of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ which can be phosphorylated by reaction with [γ-³²P]ATP. A second band contains a glycoprotein which displays an apparent molecular weight of 51 000 and thus appears to be the β subunit of the enzyme. The enzyme is sensitive to ouabain with the $\text{I}{}_{50}$ for $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ and $\text{p-}\text{nitrophenylphosphatase}$ inhibition being 1.2 and 1.3 μM, respectively. Several agents which inhibit $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from other tissues such as oligomycin, Ca²⁺, vanadate, $\text{N-}\text{ethylmaleimide}$, $\text{p-}\text{chloromercuribenzenesulfonic acid}$ (PCMBS) and 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) also inhibit the lens enzyme. Monovalent cations other than K⁺ are partially effective in activating the ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}$)-ATPase and $\text{p-}\text{nitrophenylphosphatase}$ activities. The K⁺ congeners were relatively more effective in supporting $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ compared to $\text{p-}\text{nitrophenylphosphatase}$ activity. Other kinetic properties of the lens enzyme are also comparable to those of the enzyme from other tissues. Utilizing the partially purified membrane bound enzyme, discontinuities in Arrhenius plots of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity, $\text{p-}\text{nitrophenylphosphatase}$ activity and fluoresence polarization of the fluidity probe, 1,6-diphenyl-1,3,5-hexatriene (DPH), are observed near the physiological temperature of lens. The possible significance of these observations for the mechanism of cataract formation are discussed.     

An electron spin probe study of (Na+ + K+)-ATPase-Containing membranes

The modulating effect of membrane lipids on enzyme function has been described by several investigators. We have used the spin probe $\text{N-}\text{oxyl}\text{-4′,4′-}\text{dimethyloxazolidine}\text{-12-}\text{keto}$ methyl stearate (M 12-NSE) to study this interaction in ox brain membranes enriched with $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$. This methyl ester of stearic acid is practically insoluble in aqueous media, and consequently spectra of M 12-NSE-labelled preparations are free of “liquid lines”.At least two types of spectra may be obtained when ox brain microsomes are spin labelled with M 12-NSE, indicating the presence of two distinct binding sites. At one site the spin label is relatively unrestricted and gives rise to an isotropic spectrum. A second spectrum, which is obtained from spin label at another site, is similar to that which is observed after incorporation of M 12-NSE into phospholipid bilayers. This suggests that this latter site is within the core of the microsomal membrane.The two binding sites differ in their affinity for the spin probe. The low affinity site is both more abundant in crude preparations and is more easily removed by detergent treatment; spin labels at this site produce isotropic spectra. The high affinity sites are fewer in number and produce broad spectra. In addition these high affinity sites increase in concentration as the enzyme undergoes purification.The two sites are quite distinct in their sensitivity to ascorbic acid, the low affinity site showing a considerably greater rate of reduction by this agent.This study also demonstrates that the delipidation effects of sodium dodecyl sulfate and sodium deoxycholate on $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase-enriched}$ microsomes from ox brain are not identical.It is suggested that the two spin probe binding sites represent two different lipid domains, one of which is very closely associated with the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme and may reflect a protein-directed phospholipid specificity for this enzyme.     

Annular and non-annular binding sites on the (Ca2+ + Mg2+)-ATPase

Quenching of the fluorescence of the $\text{(}\text{Ca}{}^{\mathrm{2+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{)-}\text{ATPase}$ purified from muscle sarcoplasmic reticulum can be used to measure relative binding constants of hydrophobic compounds to the phospholipid-protein interface. We show that the binding constant for cholesterol is considerably less than that for phosphatidylcholine, so that cholesterol is effectively excluded from the phospholipid annulus around the ATPase. However, dibromocholestan-3β-ol causes quenching of the fluorescence of the ATPase, and so has access to other, non-annular sites. We suggest that these non-annular sites could be at protein/protein interfaces in ATPase oligomers. Oleic acid can bind at the phospholipid/protein interface, although its binding constant is less than that for a phosphatidylcholine, and it can also bind at the postulated non-annular sites. The effects of these compounds on the activity of the ATPase depend on the structure of the phospholipid present in the systems.     

Comparison of parameters estimated from A/Ci and A/Cc curve analysis

The parameters estimated from traditional A/C _i curve analysis are dependent upon some underlying assumptions that substomatal CO₂ concentration (C _i) equals the chloroplast CO₂ concentration (C _c) and the C _i value at which the A/C _i curve switches between Rubisco- and electron transport-limited portions of the curve (C _i-t) is set to a constant. However, the assumptions reduced the accuracy of parameter estimation significantly without taking the influence of C _i-t value and mesophyll conductance (g _m) on parameters into account. Based on the analysis of Larix gmelinii’s A/C _i curves, it showed the C _i-t value varied significantly, ranging from 24 Pa to 72 Pa and averaging 38 Pa. t-test demonstrated there were significant differences in parameters respectively estimated from A/C _i and A/C _c curve analysis (p<0.01). Compared with the maximum ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation rate (V_cmax), the maximum electron transport rate (J_max) and J_max/V_cmax estimated from A/C _c curve analysis which considers the effects of g _m limit and simultaneously fits parameters with the whole A/C _c curve, mean V_cmax estimated from A/C _i curve analysis (V_cmax-C _i) was underestimated by 37.49%; mean Jmax estimated from A/C _i curve analysis (J_max-C _i) was overestimated by 17.8% and (J_max-C _i)/(V_cmax-C _i) was overestimated by 24.2%. However, there was a significant linear relationship between V_cmax estimated from A/C _i curve analysis and V_cmax estimated from A/C _c curve analysis, so was it J_max (p<0.05).     

A/California/7/2009与A/California/4/2009病毒感染力比较

目的甲型H1N1流感病毒A/California/7/2009与A/California/4/2009病毒序列比较同源性在99%以上,本实验旨在比较两株病毒感染BALB/c小鼠研究感染力强弱。方法分别将A/California/7/2009（CA7）与A/California/4/2009（CA4）两株病毒分别连续10倍稀释后,对4~6周龄雌性BALB/c小鼠经乙醚麻醉后进行滴鼻攻毒,每个稀释度接种10只实验小鼠,测定CA7 MLD50为101.24/0.05 mL,检测小鼠感染、致病的多项指标,观察期为14 d。结果相同TCID50的CA7和CA4病毒感染小鼠,CA4感染小鼠后14 d内死亡率为20%,而CA7感染小鼠后8 d内死亡率为100%。CA7 106TCID50感染的小鼠病理表现为重度弥漫性间质性肺炎,CA4 106TCID50感染的小鼠病理表现为中度-重度间质性肺炎。结论在相同条件下,CA7感染力明显强于CA4。     

Comparative study of Bifidobacteriumanimalis, Escherichiacoli, Lactobacilluscasei and Saccharomycesboulardii probiotic properties

The present work investigates some probiotic properties of four different microorganisms (Bifidobacterium animalis var. lactis BB-12, Escherichia coli EMO, Lactobacillus casei and Saccharomyces boulardii). In vitro and in vivo tests were carried out to compare cell wall hydrophobicity, production of antagonistic substances, survival capacity in the gastrointestinal tract of germ-free mice without pathological consequence, and immune modulation by stimulation of Küpffer cells, intestinal sIgA and IL-10 levels. In vitro antagonism against pathogenic bacteria and yeast was only observed for the probiotic bacteria B. animalis and L. casei. The hydrophobic property of the cell wall was higher for B. animalis and E. coli EMO, and this property could be responsible for a better ability to colonize the gastrointestinal tract of germ-free mice. Higher levels of sIgA were observed mainly for S. boulardii, followed by E. coli EMO and B. animalis, and only S. boulardii induced a significant higher level of IL-10. In conclusion, for a probiotic use, S. boulardii presented better characteristics in terms of immunomodulation, and B. animalis and L. casei for antagonistic substance production. The knowledge of the different probiotic properties could be used to choice the better microorganism depending on the therapeutic or prophylactic application.