阿托伐他汀联合参松养心胶囊治疗老年心力衰竭患者临床疗效及安全性研究

目的:探讨阿托伐他汀联合参松养心胶囊对老年心力衰竭患者的临床疗效及安全性。方法:选取我院心血管内科治疗的老年慢性心力衰竭患者121例,按治疗药物不同分为两组,对照组46例采用西医常规治疗,研究组75例常规治疗基础上给予阿托伐他汀联合参松养心胶囊治疗,记录两组12个月后的临床疗效,治疗后3、6及12个月后的总胆固醇(Total cholesterol,TC)、甘油三酯(Triglyceride,TG)、高密度脂蛋白胆固醇(High density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol,LDL-C)水平的变化及不良反应的发生情况。结果:与治疗3个月时相比,两组治疗12个月时临床有效率均显著提高,研究组治疗6、12个月后临床有效率均显著高于对照组(85.33%vs. 76.09%, 88%vs. 80.43%,P0.05)。两组治疗后血清TC、TG、LDL-C水平逐步下降,血清HDL-C水平逐步上升,研究组治疗第12月后血清TC、TG及LDL-C水平较对照组显著降低(P0.05),血清HDL-C水平较对照组明显升高(P0.05)。两组在1年观察期内均未见明显肝肾功能损伤及肌溶解,不良反应发生率比较差异无统计学意义(P0.05)。结论:长期使用阿托伐他汀联合参松养心胶囊治疗老年性心力衰竭的临床效果明显优于西医常规治疗,其可有效降低血脂水平,安全性高。     

胰岛素局部应用对糖尿病足患者血清炎性因子、VEGF、氧化应激和创面组织β-catenin表达的影响

目的:研究胰岛素局部应用对糖尿病足(DF)患者血清炎性因子、血管内皮生长因子(VEGF)、氧化应激和创面组织β-catenin表达的影响,旨在为临床DF患者的治疗提供理论依据。方法:将2017年4月至2019年12月于我院接受诊治的90例DF患者纳入研究,按照随机信封法将其分成观察组及对照组各45例。对照组实施负压吸引(NPWT)治疗,观察组则于NPWT治疗的基础上增用胰岛素治疗。比较两组治疗前后创面面积、炎性因子、血管内皮生长因子(VEGF)水平、氧化应激和创面组织β-catenin表达变化情况。结果:治疗后观察组及对照组创面面积均少于治疗前,且观察组少于对照组(均P0.05)。治疗后观察组及对照组血清白细胞介素-1(IL-1)及肿瘤坏死因子-α(TNF-α)均低于治疗前,而VEGF水平高于治疗前(均P0.05);治疗后观察组血清IL-1及TNF-α均低于对照组,而VEGF水平高于对照组(均P0.05)。治疗后观察组及对照组丙二醛(MDA)水平低于治疗前,而超氧化物歧化酶(SOD)水平高于治疗前(均P0.05);治疗后观察组MDA水平低于对照组,而SOD水平高于对照组(均P0.05)。治疗后观察组及对照组创面组织β-catenin表达水平均高于治疗前,且观察组高于对照组(均P0.05)。结论:胰岛素局部应用可促进DF患者的创面愈合,同时有利于下调血清炎性因子水平,减轻氧化应激反应,其主要作用机制可能与上调创面组织VEGF、β-catenin表达有关。     

序贯血液净化对严重创伤合并MODS患者炎性因子以及血流动力学的影响

目的:探讨序贯血液净化(SBP)对严重创伤合并多器官功能障碍综合征(MODS)患者血清炎性因子及血流动力学的影响。方法:选取2015年7月-2017年5月第三军医大学新桥医院收治的严重创伤合并MODS患者48例,根据患者的治疗方式分为对照组21例以及观察组27例,其中对照组接受常规内科治疗,观察组在对照组的基础上进行SBP治疗,在治疗前后对两组患者的肝肾功能、血清炎性因子水平、血流动力学指标进行检测,并对比两组患者ICU住院时间、病死率、机械通气时间。结果:治疗后两组患者的总胆红素(TBIL)、直接胆红素(DBIL)、谷草转氨酶(AST)、谷丙转氨酶(ALT)、血尿素氮(BUN)、肌酐(Cr)水平均较治疗前降低,且治疗后观察组患者各指标低于对照组(P0.05);治疗后两组患者血清C反应蛋白(CRP)、白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平均较治疗前显著降低,且治疗后观察组患者各炎性因子水平显著低于对照组(P0.05);治疗后两组患者的心率(HR)、中心静脉压(CVP)较治疗前降低,平均动脉压(MAP)、心脏指数(CI)较治疗前升高,且治疗后观察组患者的HR、CVP显著低于对照组,MAP、CI显著高于对照组(P0.05);观察组患者的ICU住院时间、机械通气时间均较对照组缩短,病死率较对照组明显降低(P0.05)。结论:SBP应用于严重创伤合并MODS患者的治疗可以有效减轻患者的炎性反应,并可以显著改善患者血流动力学以及预后。     

依达拉奉联合纳美芬对急性重型颅脑损伤患者血清神经细胞因子和炎性因子的影响

目的:探讨依达拉奉联合纳美芬对急性重型颅脑损伤患者血清神经细胞因子和炎性因子的影响。方法:选择2015年1月至2017年12月我院接诊的急性重型颅脑损伤患者80例,按照随机数字表法分为对照组(n=40)和观察组(n=40),对照组患者入院后接受常规综合治疗,观察组在此基础上给予依达拉奉联合纳美芬进行治疗,比较两组患者血清神经细胞因子、炎性因子水平及格拉斯哥昏迷评分(GCS)、急性生理学及慢性健康状况Ⅱ(APACHEⅡ)评分变化情况,观察两组患者颅内压情况、脑水肿情况以及不良反应发生情况。结果:观察组患者治疗后1d、治疗后1周血清β-内咖肽、S100β蛋白、神经特异性烯醇化酶(NSE)水平均低于治疗前和对照组(P0.05)。两组患者治疗后1周血清超敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)水平均较治疗前降低,且观察组低于对照组(P0.05)。观察组患者治疗1周后颅内压15 mm Hg所占比例及轻度脑水肿所占比例均高于对照组(P0.05),而颅内压≥20 mm Hg所占比例及重度脑水肿所占比例均著低于对照组(P0.05)。两组患者治疗后1周GCS评分较治疗前升高,APACHEⅡ评分较治疗前降低(P0.05);且观察组治疗后1周GCS评分较对照组升高,APACHEⅡ评分较对照组降低(P0.05)。两组患者不良反应发生率经统计分析差异无统计学意义(P0.05)。结论:依达拉奉联合纳美芬能够明显改善急性重型颅脑损伤患者血清神经细胞因子和炎性因子水平,促进颅内压下降和脑水肿吸收,有利于提升患者预后,且安全性好。     

玻璃酸钠关节腔内填充对踝关节创伤性关节炎血清TNF-α影响

目的:探究玻璃酸钠关节腔内填充对踝关节创伤性关节炎患者TNF-α及功能恢复的影响。方法:收集2014年3月到2016年3月来我院就诊的踝关节创伤性关节炎患者96例,随机分为对照组与试验组,各48例。对照组采用基础中医推拿按摩疗法配合常规功能训练,试验组实施玻璃酸钠关节腔内充填治疗。治疗5周后,比较两组患者血清炎性因子TNF-α水平的改变、功能恢复情况及不良反应发生率。结果:治疗结束后,两组ROM评分均较治疗前升高(P0.05),血清炎性因子TNF-α水平较治疗前降低(P0.05);与对照组相比,试验组实施玻璃酸钠关节腔内充填治疗ROM评分较高(P0.05),血清炎性因子TNF-α水平较低(P0.05);试验组不良反应率明显低于对照组(P0.05)。结论:玻璃酸钠关节腔内填充对踝关节创伤性关节炎患者踝关节功能恢复效果较好,能够降低患者血清炎性因子TNF-α水平,改善关节功能。     

枯草杆菌二联活菌肠溶胶囊联合康复新液灌肠治疗溃疡性结肠炎的临床疗效

目的:探讨枯草杆菌二联活菌肠溶胶囊联合康复新液灌肠治疗溃疡性结肠炎的临床疗效。方法:选取2014年5月～2018年6月淮安市淮阴医院诊治的溃疡性结肠炎患者120例,根据患者入院先后顺序分为两组,对照组在常规治疗的基础上给予枯草杆菌二联活菌肠溶胶囊,观察组在对照组的基础上给予康复新液保留灌肠。比较两组患者的治疗总有效率、治疗前后血清白介素-8(IL-8)、白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平、血小板(PLT)和血酶原时间(PT)、纤维蛋白原(FIB)水平的变化及不良反应的发生情况。结果:治疗后,观察组总有效率为95.00%,对照组为83.33%,观察组显著高于对照组(P0.05)。两组患者治疗后血清IL-8、IL-6、TNF-α水平及PLT、FIB均较治疗前显著下降,且观察组以上指标均显著低于对照组(P0.05),而两组治疗后PT水平均较治疗前显著升高,且观察组显著高于对照组(P0.05)。两组不良反应发生率相比无统计学差异(P0.05)。结论:枯草杆菌二联活菌肠溶胶囊联合康复新液灌肠治疗可显著降低溃疡性结肠炎患者的炎性因子水平,改善凝血功能,提高临床治疗效果,且安全性较高。     

口服四种维生素联合FE复合酶含漱治疗复发性口腔溃疡的疗效及对血清炎性因子水平的影响

目的:探讨口服四种维生素[维生素E(Vit E)+叶酸(FA)+维生素B2(Vit B2)+维生素B12(Vit B12)]联合FE复合酶含漱治疗复发性口腔溃疡(ROU)的临床效果及对患者血清炎性因子水平的影响。方法:选取我院2014年1月~2016年2月收治的126例ROU患者,采用随机数字表法均分为两组。对照组予以FE复合酶治疗,观察组在此基础上口服四种维生素(Vit E+FA+Vit B2+Vit B12)治疗。记录比较两组的局部疗效、远期疗效,治疗前后血清炎性因子水平以及不良反应的发生情况。结果:与对照组相比,观察组治疗30d后疼痛指数显著降低(P0.01),平均溃疡期显著缩短(P0.01)。治疗后6个月,观察组总有效率为95.2%,较对照组明显上升(81.0%,P0.05)。与治疗前对比,两组治疗30 d后血清TNF-α、IL-17水平均显著降低(P0.01),IL-2水平均显著升高(P0.01);且观察组以上各炎性因子的改善效果均更为显著(P0.01)。两组不良反应率对比差异无统计学意义(P0.05)。结论:口服四种维生素联合FE复合酶含漱治疗复发性口腔溃疡更能有效促进溃疡创面愈合,减轻疼痛,调节机体促/抗炎因子平衡,提高远期治疗效果,且安全性高。     

加贝酯联合奥曲肽对急性胰腺炎患者胃肠功能、免疫功能及炎性因子水平的影响

目的:探讨加贝酯联合奥曲肽对急性胰腺炎(AP)患者胃肠功能、血清炎性因子及免疫功能的影响。方法:选取2014年1月到2018年6月在我院接受治疗的AP患者100例,采用随机数字表法将所有患者分为对照组和观察组各50例。对照组给予注射用醋酸奥曲肽治疗,观察组在对照组的基础上给予注射用甲磺酸加贝酯治疗。对比两组患者治疗后临床疗效、胃肠功能恢复情况、血清炎性因子水平及免疫球蛋白lgG、lgM、lgA的变化情况。结果:治疗后观察组总有效率为83.24%,高于对照组的67.03%(P0.05)。治疗后观察组排气恢复时间、排便恢复时间、肠鸣音消失时间、腹胀缓解时间、腹痛缓解时间均显著短于对照组(P0.05)。治疗后两组肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)、白细胞介素-6(IL-6)及白细胞介素-8(IL-8)水平较治疗前均明显下降(P0.05),且治疗后观察组的TNF-α、CRP、IL-6及IL-8水平低于对照组(P0.05)。治疗后两组的lgG、lgM、lgA水平均较治疗前均明显升高(P0.05),治疗后观察组的lgG、lgM水平高于对照组(P0.05)。结论:加贝酯联合奥曲肽治疗AP不仅能够有效抑制患者的炎症反应,改善其免疫功能,还能够促进患者胃肠功能恢复,提高临床疗效。     

依达拉奉联合阿替普酶对急性脑梗死炎性因子的影响

目的:探讨依达拉奉联合阿替普酶治疗急性脑梗死的临床疗效以及对炎性因子水平的影响。方法:收取2012年5月至2015年5月间我院收治的急性脑梗死患者115例作为研究对象,随机分为观察组及对照组。对照组57例仅给予阿替普酶治疗,观察组58例在对照组的基础上加用依达拉奉。比较两组患者炎性因子水平、神经功能缺损评分、临床疗效以及不良反应发生情况。结果:治疗前,两组患者CRP、IL-6、IL-8及TNF-α等各项炎性因子水平以及NIHSS评分均无显著差异(P0.05)。两组患者炎性因子水平随治疗进行显著下降,观察组显著低于对照组(P0.05)。治疗后3个月,观察组患者总有效率(96.55%)显著高于对照组(85.96%),差异有统计学意义(P0.05)。观察组不良反应发生率(5.17%)与对照组(7.02%)相比无显著差异(P0.05)。结论:依达拉奉联合阿替普酶可有效降低血清炎症因子水平,治疗急性脑梗死具有良好的临床疗效及安全性,值得临床推广应用。     

黄葵胶囊联合组合型人工肾对糖尿病肾病维持性血液透析患者炎性因子、氧化应激及生活质量的影响

摘要 目的：探讨黄葵胶囊联合组合型人工肾对糖尿病肾病维持性血液透析患者炎性因子、氧化应激及生活质量的影响。方法：选取2018年10月~2019年9月我院收治的120例终末期糖尿病肾病维持性血液透析患者,将其按照随机数字表法分为观察组和对照组,各60例。两组患者均给予基础治疗,对照组采取组合型人工肾治疗。观察组在对照组基础上给予黄葵胶囊治疗。比较两组治疗3个月后的肾功能指标、炎性因子水平、氧化应激指标水平及生活质量,并统计不良反应发生情况。结果：治疗3个月后,两组血清尿素氮(BUN)、血清肌酐(Scr)、白蛋白排泄率(UAER)、超敏-C反应蛋白(hs-CRP)、白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）、晚期氧化蛋白产物（AOPP）、丙二醛（MDA）水平低于治疗前,且观察组低于对照组（P＜0.05）。两组过氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-PX）水平及SF-36各维度评分均高于治疗前,且观察组高于对照组（P＜0.05）。两组不良反应发生率比较无明显差异（P＞0.05）。结论：黄葵胶囊联合组合型人工肾治疗终末期糖尿病肾病可有效减轻患者氧化应激及炎症反应,改善患者肾功能及生活质量,安全性高,具有较好的临床应用价值。     

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.     

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。     

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).     

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.