阿奇霉素序贯治疗联合麻杏石甘汤加味对痰热闭肺型肺炎支原体肺炎患儿Th17/Treg细胞因子和NLRP3炎症小体通路的影响

摘要 目的：观察阿奇霉素序贯治疗联合麻杏石甘汤加味对痰热闭肺型肺炎支原体肺炎（MPP）患儿辅助性T细胞17（Th17）/调节性T细胞（Treg）细胞因子和NOD样受体蛋白3（NLRP3）炎症小体通路的影响。方法：选择山东省中医院2019年4月~2021年5月期间收治的痰热闭肺型MPP患儿106例,根据随机数字表法分为对照组和研究组,各为53例,对照组患儿接受阿奇霉素序贯治疗,研究组患儿接受阿奇霉素序贯治疗联合麻杏石甘汤加味治疗,对比两组疗效、中医证候积分、Th17/Treg细胞因子和NLRP3炎症小体通路相关指标,记录两组不良反应发生情况。结果：与对照组相比,研究组的临床总有效率明显更高（P<0.05）。两组治疗后次证积分、主证积分、总积分均较治疗前下降,且研究组低于对照组（P<0.05）。两组治疗后白介素-17A（IL-17A）、白介素-25（IL-25）水平均较治疗前下降,白介素-10（IL-10）、白介素-35（IL-35）水平均较治疗前升高,且研究组的变化程度大于对照组（P<0.05）。两组治疗后NLRP3 mRNA、接头蛋白凋亡相关斑点样蛋白（ASC）mRNA、半胱天冬酶1（caspase-1） mRNA均较治疗前下降,且研究组的下降程度大于对照组（P<0.05）。两组在治疗期间均未曾出现明显不良反应。结论：阿奇霉素序贯治疗联合麻杏石甘汤加味治疗痰热闭肺型MPP患儿疗效显著,可促进症状改善,作用机制可能与调节Th17/Treg细胞因子和NLRP3炎症小体通路有关。     

利拉鲁肽联合达格列净对超重或肥胖2型糖尿病患者肾功能、氧化应激以及内脏脂肪含量的影响

摘要 目的：探讨利拉鲁肽联合达格列净对超重或肥胖2型糖尿病（T2DM）患者肾功能、氧化应激以及内脏脂肪含量的影响。方法：选取我院于2018年1月~2020年5月期间接收的108例超重或肥胖T2DM患者,按照随机数字表法分为对照组（n=54）和观察组（n=54）。对照组给予利拉鲁肽治疗,观察组给予利拉鲁肽联合达格列净治疗,均治疗12周。对比两组肾功能[血胱抑素 C（CysC）、血肌酐（Scr）、血尿酸（SUA）]、氧化应激[丙二醛（MDA）、谷胱甘肽过氧化物酶（GSH-PX）、超氧化物歧化酶（SOD）]、体质量指数（BMI）、腰围、血糖指标[空腹血糖（FBG）、餐后2 h血糖（2hPBG）、糖化血红蛋白（HbA1c）]以及体成分指标（全身脂肪百分比、内脏脂肪含量）,记录两组治疗期间不良反应情况。结果：两组治疗后BMI、腰围、2hPBG、FBG、HbA1c均下降,且观察组较对照组低（P<0.05）。两组治疗后MDA均下降,且观察组较对照组低（P<0.05）,两组治疗后SOD、GSH-PX均升高,且观察组较对照组高（P<0.05）。两组治疗后全身脂肪百分比、内脏脂肪含量均下降,且观察组较对照组低（P<0.05）。两组治疗前后CysC、Scr、SUA组内及组间对比均无统计学差异（P>0.05）。两组不良反应发生率对比无统计学差异（P>0.05）。结论：超重或肥胖T2DM患者利拉鲁肽治疗基础上联合达格列净,降糖效果确切,减轻机体氧化应激,降低内脏脂肪含量,对肾功能无显著影响,且不增加不良反应发生率。     

茵陈五苓散联合利拉鲁肽对肥胖型2型糖尿病患者糖脂代谢、胰岛素敏感性和氧化应激的影响

摘要 目的：探讨茵陈五苓散联合利拉鲁肽对肥胖型2型糖尿病（T2DM）患者糖脂代谢、胰岛素敏感性和氧化应激的影响。方法：选取2021年12月~2022年12月期间广州中医药大学附属佛山中医院收治的160例痰湿内蕴型肥胖T2DM患者。根据随机数字表法将患者分为对照组（利拉鲁肽治疗,80例）和研究组（茵陈五苓散联合利拉鲁肽治疗,80例）。对比两组疗效、糖脂代谢指标、肥胖相关指标、胰岛素敏感性、氧化应激和不良反应发生情况。结果：与对照组相比,研究组的临床总有效率更高（P<0.05）。研究组治疗后体质量指数（BMI）、空腹血糖（FBG）、腰臀比（WHR）、餐后2 h血糖（2hPG）、丙二醛（MDA）、糖化血红蛋白（HbAlc）、稳态模型胰岛素抵抗指数（HOMA-IR）、总胆固醇（TC）、黄嘌呤氧化酶（XO）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）较对照组低（P<0.05）。治疗后研究组稳态模型胰岛β细胞功能指数（HOMA-β）、高密度脂蛋白胆固醇（HDL-C）、超氧化物歧化酶（SOD）高于对照组（P<0.05）。两组不良反应发生率组间对比未见差异（P>0.05）。结论：茵陈五苓散联合利拉鲁肽治疗肥胖型T2DM患者疗效确切,可调节糖脂代谢水平,降低胰岛素敏感性,减轻氧化应激,且具有一定安全性,值得临床借鉴应用。     

参元益气活血胶囊联合心痛贴穴位贴敷对不稳定性心绞痛气虚血瘀证患者脂代谢和氧化应激的影响

摘要 目的：观察参元益气活血胶囊联合心痛贴穴位贴敷对不稳定性心绞痛（UA）气虚血瘀证患者脂代谢和氧化应激的影响。方法：按照随机数字表法将2020年3月~2022年9月期间首都医科大学附属北京中医医院收治的100例UA患者分为对照组（常规西医治疗和心痛贴穴位贴敷）和研究组（对照组的基础上接受参元益气活血胶囊治疗），各为50例。对比两组中医证候积分、心绞痛发作次数和持续时间、脂代谢指标[总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）]、氧化应激指标[超氧化物歧化酶（SOD）、丙二醛（MDA）]、心功能指标[左心室射血分数（LVEF）、心输出量（CO）、左心室舒张末期内径（LVEDD）]。结果：两组治疗4周后胸痛、胸闷、气短、心悸、神倦乏力、自汗、不寐评分均下降，且研究组均低于对照组（P<0.05）。两组治疗4周后心绞痛发作次数减少，持续时间缩短，且研究组改善幅度大于对照组（P<0.05）。两组治疗4周后TG、TC、LDL-C下降，HDL-C升高，且研究组改善幅度大于对照组（P<0.05）。两组治疗4周后SOD升高，MDA下降，且研究组改善幅度大于对照组（P<0.05）。两组治疗4周后LVEF、CO升高，LVEDD下降，且研究组改善幅度大于对照组（P<0.05）。结论：参元益气活血胶囊联合心痛贴穴位贴敷治疗UA气虚血瘀证患者，可减少心绞痛发作次数，缩短持续时间，促进临床症状改善，改善心功能、脂代谢和氧化应激水平。     

加味左归丸联合雌孕激素序贯治疗对早发性卵巢功能不全患者子宫动脉血流动力学、氧化应激和免疫因子的影响

摘要 目的：探讨加味左归丸联合雌孕激素序贯治疗对早发性卵巢功能不全（POI）患者子宫动脉血流动力学、氧化应激和免疫因子的影响。方法：纳入我院2019年10月-2021年10月期间收治的100例POI患者,按照随机数字表法分为对照组（雌孕激素序贯治疗,50例）和研究组（加味左归丸联合雌孕激素序贯治疗,50例）。对比两组中医证候积分、子宫动脉血流动力学、氧化应激指标、性激素指标和免疫因子指标。结果：两组治疗后月经周期、月经量、腰膝酸软、潮热盗汗、头晕耳鸣、阴道干涩、失眠多梦、五心烦热、性欲降低评分均下降,且研究组低于对照组（P<0.05）。两组治疗后子宫动脉收缩期峰值流速（Vmax）升高,搏动指数（PI）、阻力指数（RI）下降,且研究组的变化程度大于对照组（P<0.05）。两组治疗后血清丙二醛（MDA）下降,谷胱甘肽过氧化物酶（GSH-Px）、超氧化物歧化酶（SOD）升高,且研究组的变化程度大于对照组（P<0.05）。两组治疗后雌二醇（E2）升高,促黄体生成素（LH）、促卵泡生成激素（FSH）下降,且研究组的变化程度大于对照组（P<0.05）。两组治疗后NK细胞、CD8⁺下降,CD3⁺、CD4⁺、CD4⁺/CD8⁺升高,且研究组的变化程度大于对照组（P<0.05）。结论：加味左归丸联合雌孕激素序贯治疗POI患者,可有效改善子宫动脉血流动力学、性激素、氧化应激和免疫功能,促进患者恢复。     

重复经颅磁刺激对脑卒中后认知功能障碍患者神经功能指标和外周血NLRP3炎症小体的影响

摘要 目的：探讨重复经颅磁刺激（rTMS）对脑卒中后认知功能障碍（PSCI）患者神经功能指标和外周血Nod样受体蛋白3（NLRP3）炎症小体的影响。方法：选择2020年4月~2022年5月期间广东三九脑科医院收治的PSCI患者98例,按照随机数字表法将患者分为对照组（常规药物治疗及康复训练,n=49）和实验组（对照组基础上接受rTMS,n=49）。对比两组量表评分、神经功能指标、外周血NLRP3炎症小体。结果：治疗4周后,实验组简易智能状态检查量表（MMSE）、蒙特利尔认知评估量表（MoCA）、改良Barthel指数量表（MBI）评分高于对照组（P<0.05）。实验组胶质纤维酸性蛋白（GFAP）、神经元特异性烯醇化酶（NSE）、S100β低于对照组（P<0.05）。实验组外周血NLRP3mRNA、白细胞介素-18（IL-18）、白细胞介素-1β（IL-1β）低于对照组（P<0.05）。结论：rTMS治疗PSCI,可有效减轻患者认知功能障碍,提高生活自理能力,调节患者的神经功能指标和外周血NLRP3炎症小体。     

五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎患者氧化应激、CD4+CD25+调节性T细胞和血清MMP-1、MMP-2、TIMP-1的影响

摘要 目的：探讨五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎（CHB）患者氧化应激、CD4⁺CD25⁺调节性T细胞和血清基质金属蛋白酶-1（MMP-1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶组织抑制因子-1（TIMP-1）的影响。方法：纳入苏州大学附属传染病医院2021年6月~2022年12月期间收治的CHB患者122例,采用随机数字表法分为对照组（n=61,富马酸丙酚替诺福韦片）和研究组（n=61,五灵胶囊联合富马酸丙酚替诺福韦片）。对比两组疗效、氧化应激指标、CD4⁺CD25⁺调节性T细胞、肝功能指标[总胆红素（TBIL）、谷丙转氨酶（ALT）、谷氨酰转肽酶（GGT）]、血清MMP-1、MMP-2、TIMP-1水平和不良反应发生情况。结果：研究组的临床总有效率高于对照组（P<0.05）。两组治疗后丙二醛（MDA）下降,且研究组低于对照组同时间点;超氧化物歧化酶（SOD）升高,且研究组高于对照组同时间点（P<0.05）。两组治疗后CD4⁺CD25⁺调节性T细胞下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后MMP-1、MMP-2、TIMP-1下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后TBIL、ALT、GGT下降,且研究组低于对照组同时间点（P<0.05）。两组不良反应总发生率组间对比未见差异（P>0.05）。结论：五灵胶囊联合富马酸丙酚替诺福韦片治疗CHB患者,可有效减轻机体氧化应激,调节CD4⁺CD25⁺调节性T细胞和血清MMP-1、MMP-2、TIMP-1水平。     

溶栓胶囊联合丁苯酞软胶囊对脑梗死恢复期患者血液流变学、脑血流动力学和神经因子的影响

摘要 目的：探讨溶栓胶囊联合丁苯酞软胶囊对脑梗死恢复期患者血液流变学、脑血流动力学和神经因子的影响。方法：选取2021年1月-2022年1月期间长治医学院附属和平医院收治的100例脑梗死恢复期患者,按照随机数字表法分为研究组（50例）和对照组（50例）,对照组接受丁苯酞软胶囊治疗,研究组接受溶栓胶囊联合丁苯酞软胶囊治疗。对比两组疗效、血液流变学、脑血流动力学、神经因子和不良反应发生情况。结果：研究组90.00%的临床总有效率高于对照组72.00%（P<0.05）。治疗20 d后,两组美国国立卫生院神经功能缺损评分（NIHSS）评分下降,Barthel指数（BI）评分升高,且研究组改善幅度大于对照组（P<0.05）。治疗20 d后,两组全血粘度、血浆粘度、血沉（ESR）、纤维蛋白原（FIB）和红细胞压积（HCT）均下降,且研究组低于对照组（P<0.05）。治疗20 d后,两组平均血流速度（Vm）升高,搏动指数（PI）及阻力指数（RI）均下降,且研究组改善幅度大于对照组（P<0.05）。治疗20 d后,两组神经元特异性烯醇化酶（NSE）下降,神经生长因子（NGF）、神经营养因子（BDNF）升高,且研究组改善幅度大于对照组（P<0.05）。两组不良反应发生率组间对比无差异（P>0.05）。结论：溶栓胶囊联合丁苯酞软胶囊可有效改善脑梗死恢复期患者血液流变学、脑血流动力学,促进神经功能恢复,提高生活活动能力,效果显著。     

醒脑静联合纳洛酮对急性脑出血伴意识障碍患者神经功能、炎性因子和氧化应激的影响

摘要 目的：探讨纳洛酮联合醒脑静对急性脑出血（ACH）伴意识障碍患者炎性因子、神经功能和氧化应激的影响。方法：选取2017年2月~2019年12月期间我院收治的95例ACH伴意识障碍患者,按照随机数字表法将上述患者分为对照组（n=47）和研究组（n=48）,对照组患者予以纳洛酮治疗,研究组则在对照组的基础上联合醒脑静治疗,比较两组患者疗效、神经功能、炎性因子和氧化应激指标,记录两组不良反应发生情况。结果：研究组治疗10 d后的临床总有效率为91.67%（44/48）,高于对照组的74.47%（35/47）（P<0.05）。两组治疗10 d后美国国立卫生研究院卒中量表（NIHSS）评分,白介素-6（IL-6）、C反应蛋白（CRP）以及肿瘤坏死因子-α（TNF-α）、丙二醛（MDA）水平均较治疗前下降,且研究组低于对照组（P<0.05）。两组治疗10 d后超氧歧化酶 (SOD)水平均较治疗前升高,且研究组高于对照组（P<0.05）。两组不良反应发生率对比未见统计学差异（P>0.05）。结论：醒脑静联合纳洛酮治疗ACH伴意识障碍患者,疗效显著,可有效改善患者神经功能、炎性因子和氧化应激,且安全性较好。     

益肾活血解毒汤治疗肾虚血瘀型颈动脉粥样硬化患者的疗效观察

摘要 目的：观察益肾活血解毒汤治疗肾虚血瘀型颈动脉粥样硬化（CAS）效果。方法：选取我院2021年8月~2023年4月收治的肾虚血瘀型CAS患者80例,随机分为观察组和对照组,各40例。两组均予以常规西医治疗,观察组加服益肾活血解毒汤,治疗周期为四周。观察两组的临床疗效、血脂等指标变化,并进行统计学比较。结果：两组治疗前中医证候积分无差异（P＞0.05）,而治疗后,观察组中医证候积分明显低于对照组（P<0.05）;观察组总有效率（97.50%）虽在数值上高于对照组（92.50%）,但并无统计学意义（P＞0.05）;相较于治疗前,两组治疗后血脂指标（CHOL、TG、LDL-C）和炎性因子（NLRP3、Caspase-1、IL-1β、IL-18）水平显著下降（P<0.05）,而观察组下降幅度更大,与对照组差异显著（P<0.05）;服药期间,两组均无明显不良反应发生。结论：常规调脂抗动脉粥样硬化联合益肾活血解毒汤治疗肾虚血瘀型CAS疗效肯定,可有效改善临床症状,调节血脂水平,控制炎症反应,且安全性良好。     

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.     

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.     

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.     

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).