四川辖曼自然保护区硬刺高原鳅生物学研究

对四川若尔盖辖曼自然保区220尾硬刺高原鳅的生物学进行了研究。硬刺高原鳅体长22.56～98.56mm,体重0.12～13.98g;体重与体长呈显著的幂函数关系:W=0.0095L3.2033(r=0.9956)。Fulton肥满度雌体平均为1.35,雄体平均为1.37;Clark肥满度雌体平均为1.03,雄体平均为1.05。雄性成熟个体最小体长为74.61mm,体重为5.79g。雌性成熟个体最小体长为71.06mm,体重为5.20g。平均绝对怀卵量5175粒/尾。食性主要为摇蚊幼虫,其次是藻类和维管植物。     

淮河上游南湾湖麦穗鱼年龄与生长的研究

2014年的3月、5月、7月和12月在淮河上游南湾湖采集麦穗鱼(Pseudorasbora parva)样本532尾, 对麦穗鱼的年龄组成与生长进行分析。结果表明样本的体长分布范围为35.82—88.28 mm, 平均体长为(61.61±11.8) mm, 体重的分布范围为3.07—59.17 g, 平均体重为(19.23±10.73) g。雄性个体比雌性个体大, 雌雄性比为0.64鲶1。群体的年龄组成为1—3龄, 其中3龄样本数量占优势为57.38%。体长与体重的关系是雌性W=9.602E–5L2.928 (R2=0.883); 雄性W=4.487E–5L3.116 (R2=0.889), 雌雄样本间存在显著性差异(F=5.241, P<0.05)。麦穗鱼的鳞径与体长之间呈线性关系, 并且雌雄样本的鳞径与体长之间的关系差异性显著(F=78.405, P<0.05)。生长参数分别是雌性: L_∞=107.005, K=0.246, t₀= –0.76; 雄性: L_∞=145.254, K=0.181, t₀= –0.66。生长拐点是雌性3.607龄对应的体长和体重分别为70.46 mm和24.72 g, 雄性5.619龄对应的体长和体重分别为98.64 mm和73.53 g。研究结果表明雌性为匀速生长, 雄性为异速生长; 雄性麦穗鱼比雌性麦穗鱼的生长速度快。     

北江间（鱼骨）的年龄与生长特征

于2007年4月至2009年3月间,在广东北江中上游地区采集间(鱼骨)(Hemibarbus medius)样本640尾,以鳞片作为年龄鉴定材料,对其种群的年龄与生长特征进行了研究.结果表明,间(鱼骨)鳞片上年轮结构清晰,前区主要表现为疏密型,而侧区主要为切割型;鳞片的边缘生长率表明,间(鱼骨)年轮的形成期主要为3～4月份.样本由0+～6+龄7个龄组组成,其中,雌性由1+～6+龄6个龄组组成,主要为1+～4+龄;而雄性由1+～5+龄5个龄组组成,主要为1+～3+龄;在4+ ～6+龄个体中,雌性78尾,雄性27尾,雌性显著多于雄性.体长(L,cm)与体重(W,g)的关系为W=0.011L3.149(♀)、W=0.011L3.135(♂),协方差分析表明,雌雄个体体长与体重关系的差异不显著,所有样本体长与体重的关系式为W=0.011L3.148.体长与鳞长(R,mm)呈幂函数及直线关系,关系式分别为L=6.387R0.853、L=4.569R+ 2.587.拟合的von Bertalanffy生长参数为L∞=29.855 0 cm,k=0.223 1,W∞=483.889 8 g,t0=-0.928 2龄.对理论体长与实测体长、理论体重与实测体重进行x2检验,两者的差异均不显著,表明von Bertalanffy生长方程能较好地拟合间(鱼骨)的生长.体重生长的拐点年龄为4.21龄,拐点体长约为20.37 cm,体重约为166.19 g.作为渔业保护对策,建议起捕年龄3龄以上或体长17 cm以上的个体.     

金沙江攀枝花江段棒花鱼的生物学

在2006年5月和2006年12月至2007年5月期间,通过使用三层定置刺网捕捉棒花鱼,我们研究了金沙江中游攀枝花江段棒花鱼的生物学。棒花鱼在三层定置刺网中的平均出现率为93.1%,在总渔获物中的平均重量百分比为7.68%;雄性个体的平均体长显著大于雌性个体,体长和体重的回归方程为:W=4×10-5L2.8499(W为体重,L为体长,R2=0.8614);根据鳞片年轮对82尾标本进行了年龄鉴定,其中0龄个体1尾,占1.22%;1龄个体68尾,占82.93%;2龄个体13尾,占15.85%。鳞径与体长显著相关,且雄性个体的线性回归关系比雌性个体强。绝对怀卵量为737-4516粒,相对怀卵量为180-597粒/g。棒花鱼的卵径分布范围为0.28-1.24mm,卵径分布呈双峰型。综合分析棒花鱼的性成熟系数变化趋势、繁殖期的持续时间、雌性个体的性腺发育和卵子的不同步发育,得知棒花鱼为一次性产卵鱼类。     

塔里木河叶尔羌高原鳅繁殖生物学研究

2010年7月至2011年12月,在塔里木河阿拉尔段采集叶尔羌高原鳅Triplophysa (Hedinichthys) yar-kandensis (Day)940尾(除性别未辨个体外)用于繁殖生物学研究。种群雌雄比为0.85︰1,最小性成熟,雌性个体体长为8.2 cm,体重为7.4 g,年龄为3龄;雄性个体体长为6.5 cm,体重为3.4 g,年龄为2龄。叶尔羌高原鳅卵径分布呈单峰形,推测应属于同步产卵类型。计算了88尾Ⅳ-Ⅴ期雌鱼的怀卵量,其体长范围30-195 mm,体重范围3.59-114.04 g,绝对繁殖力为1101-56320(9944±5487)粒,相对繁殖力为824-1140(982±158);塔里木河阿拉尔段叶尔羌高原鳅种群繁殖力(Fp)为403.46万粒。     

北江间的年龄与生长特征

于2007年4月至2009年3月间,在广东北江中上游地区采集间(Hemibarbus medius)样本640尾,以鳞片作为年龄鉴定材料,对其种群的年龄与生长特征进行了研究。结果表明,间鳞片上年轮结构清晰,前区主要表现为疏密型,而侧区主要为切割型;鳞片的边缘生长率表明,间年轮的形成期主要为3~4月份。样本由0+~6+龄7个龄组组成,其中,雌性由1+~6+龄6个龄组组成,主要为1+~4+龄;而雄性由1+~5+龄5个龄组组成,主要为1+~3+龄;在4+~6+龄个体中,雌性78尾,雄性27尾,雌性显著多于雄性。体长(L,cm)与体重(W,g)的关系为W=0.011L3.149(♀)、W=0.011L3.135(♂),协方差分析表明,雌雄个体体长与体重关系的差异不显著,所有样本体长与体重的关系式为W=0.011L3.148。体长与鳞长(R,mm)呈幂函数及直线关系,关系式分别为L=6.387R0.853、L=4.569R+2.587。拟合的von Bertalanffy生长参数为L∞=29.855 0 cm,k=0.223 1,W∞=483.889 8 g,t0=﹣0.928 2龄。对理论体长与实测体长、理论体重与实测体重进行χ~2检验,两者的差异均不显著,表明von Bertalanffy生长方程能较好地拟合间的生长。体重生长的拐点年龄为4.21龄,拐点体长约为20.37 cm,体重约为166.19 g。作为渔业保护对策,建议起捕年龄3龄以上或体长17 cm以上的个体。     

鄱阳湖黄鳝的生长特征

以基舌骨和脊椎骨作为年龄鉴定的材料,研究了鄱阳湖黄鳝(Monopterus albus)种群的生长特征.结果表明,种群的年龄结构分别为:雌性1～5龄,雄性2～6龄,2～5龄为黄鳝性别的过渡期(间性).2～4龄为优势年龄组,占渔获物的89.75%,相对应的体长为30～50 cm,体重为30～120 g.体长(L)与体重(W)的关系为:W=0.000 4L3.2601(♀);W=0.001 4L2.9008(∮).按生长指标值分析,阶段生长可以明显地划分为两个时期,即2龄前的生长迅速期和2龄后的生长稳定期.拟合的Von Bertalanffy生长参数分别为雌性L∞=78.5 Cm,k=0.174 02/y,t0=-1.203 2 Y,W∞:602.01 g;雄性L∞=102.3 cm,k=0.118 45/y,t0=-1.310 1 Y,W-=947.32 g;生长特征参数分别为ψ=3.030 3(♀)和ψ=3.093 4(∮);雌雄个体生长差异显著.     

青衣江下游中华沙鳅繁殖生物学的初步研究

2013年5月-2014年6月,对青衣江下游夹江至乐山段中华沙鳅的资源现状,繁殖生物学进行了调查,并采用常规方法对73尾繁殖期野生中华沙鳅进行了观察研究,结果:73尾中华沙鳅标本,其体长在84~184mm范围,平均体长为134mm;体重在10.87~45.57g之间,平均体重为22.64g;其中31尾雌性中华沙鳅绝对怀卵量1542~7563粒,平均怀卵量为4486粒;雌雄中华沙鳅初次性成熟年龄均为2龄。     

北江侧条光唇鱼的年龄与生长特征

于2007年7月至2009年6月间,在北江中上游地区采集了358尾侧条光唇鱼(Acrossocheilus parallens)样本,以鳞片作为年龄鉴定材料,对其年龄与生长特征进行了研究。侧条光唇鱼鳞片上年轮结构清晰,前区主要为疏密型,而侧区主要为切割型;鳞片的边缘生长率显示,其年轮的主要形成期为2~3月份。样本中的雌性由1+~5+龄5个龄组组成,优势龄组为1+~3+龄;而雄性由1+~4+龄4个龄组组成,优势龄组为1+~2+龄。雌鱼的体长(L,cm)范围为5.7~14.8 cm,集中于7.0~13.0 cm之间;体重(W,g)范围为4.75~108.40 g,集中于10.00~30.00 g之间。雄鱼的体长范围为5.7~12.6 cm,集中于7.0~11.0 cm之间;体重范围为4.40~61.50 g,主要集中于10.00~20.00 g之间。体长与体重的关系为W=0.017L3.167、W♀=0.017L3.198、W♂=0.023L3.025,协方差分析表明,雌、雄个体在体长与体重关系上,差异显著。体长(L,cm)与鳞径(R,mm)呈线性关系,L♀=3.126R+1.869、L♂=2.875R+2.152。拟合出Von Bertalanffy生长参数,雌性L∞=17.143 5 cm,k=0.270 5,W∞=150.347 4 g,t0=﹣0.614 6龄;雄性L∞=17.236 5 cm,k=0.226 9,W∞=126.468 6 g,t0=﹣0.921 5龄。雌鱼体重生长的拐点年龄为3.68龄,拐点体长约为11.8 cm,体重约为45.26 g;雄鱼的拐点年龄为3.96龄,拐点体长约为11.5 cm,体重约为37.60 g。作为渔业对策,建议起捕年龄3龄以上或体长10 cm以上的个体。     

广东北江马口鱼个体生殖力研究

于2009年3、4月,从北江中随机采集成熟度为Ⅳ期的马口鱼雌性个体55尾,测量体长(L)、体重(W)、净体重(Wn)和性腺重(Wo)等形态学指标,并用鳞片鉴定年龄,用重量法计数绝对生殖力(F),计算出体长相对生殖力(FL)、体重相对生殖力(FW)、成熟系数(GSI)和丰满度(K),并用5种数学模型及多元逐步回归方程拟合了马口鱼绝对生殖力与生物学指标的关系.结果表明,用于统计生殖力的马口鱼样本由1+～3+龄3个龄组组成;绝对生殖力在1486.29～12 025.78粒之间,平均为4842.42粒;体长相对生殖力在163.33～856.27粒/cm之间,平均为437.30粒/cm;体重相对生殖力在99.77～470.27粒/g之间,平均为207.29粒/g.绝对生殖力与体长、体重、净体重、性腺重、年龄呈抛物线相关,与成熟系数呈幂函数相关,与丰满度的关系不显著.多元逐步回归分析表明,绝对生殖力与体重和成熟系数密切相关,相关式为:F=-312.129+137.765W+108.715GSI(R2=0.719,n=55,F=57.089, P<0.001).     

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.     

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。     

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.     

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.     

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).