Epr Studies on the Effects of Complexation of Heme by Hemopexin upon its Reactions with Organic Peroxides

Hemopexin, a heme-binding serum glycoprotein, is thought to play an important role in the prevention of oxidative damage that may be catalysed by free heme. Through the use of EPR techniques, the generation of free radicals from organic hydroperoxides by heme and heme-hemopexin complexes, and the concomitant formation of high oxidation-state iron species has been studied; these species are implicated as causative agents in processes such as cardiovascular disease and carcinogenesis. From the rates of production of these species from both n-alkyl and branched hydroperoxides, it has been inferred that the dramatic reduction in the yield of oxidising species generated by heme upon its complexation with hemopexin arises from steric hindrance of the access of hydroperoxide to the bound heme.     

Chemiosmotic coupling of ion transport in the yeast vacuole: Its role in acidification inside organelles

Acidification inside the vacuo-lysosome systems is ubiquitous in eukaryotic organisms and essential for organelle functions. The acidification of these organelles is accomplished by proton-translocating ATPase belonging to the V-type H⁺-ATPase superfamily. However, in terms of chemiosmotic energy transduction, electrogenic proton pumping alone is not sufficient to establish and maintain those compartments inside acidic. Current studies have shown that thein situ acidification depends upon the activity of V-ATPase and vacuolar anion conductance; the latter is required for shunting a membrane potential (interior positive) generated by the positively charged proton translocation. Yeast vacuoles possess two distinct Cl^– transport systems both participating in the acidification inside the vacuole, a large acidic compartment with digestive and storage functions. These two transport systems have distinct characteristics for their kinetics of Cl^– uptake or sensitivity to a stilbene derivative. One shows linear dependence on a Cl^– concentration and is inhibited by 4,4-diisothiocyano-2,2-stilbenedisulfonic acid (DIDS). The other shows saturable kinetics with an apparentK _m for Cl^– of approximately 20 mM. Molecular mechanisms of the chemiosmotic coupling in the vacuolar ion transport and acidification inside are discussed in detail.     

Evaluation of Superoxide Scavenging Activities of Hamamelis Extract and Hamamelitannin

Hamamelitannin, which is a component of bark extract of hamamelis (Hamamelis virginior L.), was found to be a potent scavenger of superoxide anion radicals. Superoxide anion scavenging activity of the compound was evaluated by ESR-spin trap method using DMPO (5,5'-dimethyl-1-pyrroline-N-oxide) as a spin trapping agent. The IC₅₀ value (the concentration producing 50% inhibition of superoxide anion radicals) of hamamelitannin was found to be 1.38 ± 0.06 μM much lower than that of ascorbic acid (23.31 ± 2.23 μM). Supporting the superoxide scavenging activity of hamamelitannin, the compound showed both suppresive ability against depolymelization of hyaluronic acid and protective ability against cytotoxicity induced by superoxide anion radicals. Hamamelitannin increased the survival rate of fibroblast to 85.5 ± 3.3%, compared with that of control (27.2 ± 4.3%).     

Enrichment of adeno-associated virus serotype 5 full capsids by anion exchange chromatography with dual salt elution gradients

Adeno-associated virus-based gene therapies have demonstrated substantial therapeutic benefit for the treatment of genetic disorders. In manufacturing processes, viral capsids are produced with and without the encapsidated gene of interest. Capsids devoid of the gene of interest, or “empty” capsids, represent a product-related impurity. As a result, a robust and scalable method to enrich full capsids is crucial to provide patients with as much potentially active product as possible. Anion exchange chromatography has emerged as a highly utilized method for full capsid enrichment across many serotypes due to its ease of use, robustness, and scalability. However, achieving sufficient resolution between the full and empty capsids is not trivial. In this work, anion exchange chromatography was used to achieve empty and full capsid resolution for adeno-associated virus serotype 5. A salt gradient screen of multiple salts with varied valency and Hofmeister series properties was performed to determine optimal peak resolution and aggregate reduction. Dual salt effects were evaluated on the same product and process attributes to identify any synergies with the use of mixed ion gradients. The modified process provided as high as ≥75% AAV5 full capsids (≥3-fold enrichment based on the percent full in the feed stream) with near baseline separation of empty capsids and achieved an overall vector genome step yield of >65%.     

Roles of catalase and cytochrome C in hydroperoxide-dependent lipid peroxidation and chemiluminescence in rat heart and kidney mitochondria

A recent report (Radi et al., J. Biol. Chem. 266:22028–22034, 1991) showed that rat heart mitochondria contain catalase. The protective role of mitochondrial catalase was tested by exposing heart or kidney mitochondria and mitoplasts to two oxidants (H₂O₂) or tert-butyl hydroperoxide, t-BOOH), estimating lipid peroxidation (as thiobarbituric acid-reactive substances, TBARS) and overall oxidative stress (as chemiluminescence). Additional controls included heart and kidney preparations from aminotriazole-treated (catalase-depleted) rats. Both oxidants increased TBARS in catalase-free preparations to similar extents over their respective controls (between 200 to 350%). In catalase-containing preparations, H₂O₂ lipid peroxidation increased by only 40 to 96% over controls. Similar qualitative results were obtained when measuring chemiluminescence. The catalytic role of cytochrome c in mitochondrial lipid peroxidation was investigated by exposing either control or cytochrome-c-depleted kidney mitoplasts (catalase free) to either H₂O₂ or t-BOOH. Hydrogen-peroxide-dependent mitochondrial lipid peroxidation varied with cytochrome c concentrations, remaining close to controls when cytochrome c concentration decreased by 66%, even though there was no catalase present. Tert-butyl hydroperoxide-dependent lipid peroxidation was less affected by cytochrome c remaining 2.3-fold above controls under the same conditions, suggesting that organic peroxides are more likely to remain in the less polar membrane environment being decomposed by heme or nonheme iron imbedded in the inner mitochondrial membrane. Chemiluminescence was less affected by cytochrome c depletion. Comparing control and cytochrome-c-deficient mitochondria, chemiluminescence was 1.7-fold and 2.8-fold higher when control preparations were challenged with t-BOOH or H₂O₂, respectively.     

羟自由基对人红细胞磷脂酰乙醇胺脂质体相变性质的影响

研究了过氧化氢与亚铁离子体系产生的羟自由基对人红细胞膜磷脂酸乙醇胺(PE)脂质体相变性质的影响.结果表明,羟自由基导致脂质体不饱和脂肪酸链的含量明显降低和丙二醛(MDA)含量显著升高,同时其膜流动性随之下降.在室温下,羟自由基诱使PF脂质体冰冻断裂面出现脂质颗粒,说明羟自由基通过脂质过氧化作用可促进PE脂质体从脂双层转变为非双层结构.     

血清CEA、G-17、MK、ProGRP与胃癌根治术患者术后复发风险的关系研究

摘要 目的：探讨血清癌胚抗原（CEA）、胃泌素17（G-17）、中期因子（MK）、胃泌素释放肽前体（ProGRP）与胃癌根治术（RG）患者术后复发风险的关系。方法：选取2018年1月～2020年8月新疆医科大学第一附属医院普外科收治的156例胃癌患者为胃癌组,根据RG后是否复发分为复发组和无复发组,另选取同期我院52名健康体检志愿者为对照组。收集胃癌患者临床资料,采用酶联免疫吸附法检测血清CEA、G-17、MK、ProGRP水平。通过单因素和多因素Logistic回归分析RG患者术后复发的影响因素,受试者工作特征（ROC）曲线分析血清CEA、G-17、MK、ProGRP水平对RG患者术后复发的预测价值。结果：与对照组比较,胃癌组血清CEA、G-17、MK、ProGRP水平升高（P＜0.05）。随访2年,失访2例,154例RG患者术后复发率为28.57%（44/154）。多因素Logistic回归分析显示,CEA、G-17、MK、ProGRP升高、TNM分期Ⅲ期、分化程度为低分化、淋巴结转移为RG患者术后复发的独立危险因素（P＜0.05）。ROC曲线分析显示,血清CEA、G-17、MK、ProGRP水平联合预测RG患者术后复发的曲线下面积（AUC）大于CEA、G-17、MK、ProGRP单独预测。结论：血清CEA、G-17、MK、ProGRP水平升高与RG患者术后复发密切相关,可能成为RG患者术后复发的辅助预测指标,且血清CEA、G-17、MK、ProGRP联合预测RG患者术后复发的价值较高。     

乳腺癌根治术患者术前焦虑的影响因素分析及其对术后恢复、细胞免疫功能和生命质量的影响

摘要 目的：分析乳腺癌根治术患者术前焦虑的影响因素,并探讨术前焦虑对患者术后恢复、细胞免疫功能和生命质量的影响。方法：选择我院2020年3月~2021年12月期间收治的拟行乳腺癌根治术的120例患者作为研究对象,术前1 d采用焦虑自评量表（SAS）评估所有患者的焦虑状况,根据是否存在焦虑分为焦虑组和无焦虑组,乳腺癌根治术患者术前焦虑的影响因素采用多因素Logistic回归分析。对比焦虑组和无焦虑组的术后恢复、细胞免疫功能和生命质量情况。结果：120例乳腺癌根治术患者中,有31例患者无术前焦虑,89例患者存在焦虑症状,根据是否存在术前焦虑分为焦虑组（n=89）和无焦虑组（n=31）。乳腺癌根治术患者术前焦虑与年龄、文化程度、家庭人均月收入、付费方式、婚姻状况、家庭支持、既往有无全麻史、术前住院时长、定期体检有关（P<0.05）。多因素Logistic回归分析结果表明：年龄＜60岁、文化程度为小学及其以下、家庭人均月收入＜3000元、婚姻状况为未婚、无家庭支持、既往无全麻史、术前住院时长>1 d是乳腺癌根治术患者术前焦虑的危险因素（P<0.05）。焦虑组的术后首次肛门排气时间、首次下床活动时间、术后住院时间均长于无焦虑组（P<0.05）。两组术后1个月CD3⁺、CD4⁺、CD4⁺/CD8⁺水平升高,且无焦虑组高于焦虑组（P<0.05）,两组术后1个月CD8⁺水平下降,且无焦虑组低于焦虑组（P<0.05）。两组术后1个月生理状况、情感状况、社会/家庭状况、功能状况、附加关注评分和总分下降,且无焦虑组低于焦虑组（P<0.05）。结论：乳腺癌根治术患者术前焦虑发生率较高,其发生受到年龄、文化程度、家庭人均月收入、婚姻状况、家庭支持、既往有无全麻史、术前住院时长等多种因素的影响,可导致患者术后恢复时间延长,细胞免疫功能和生命质量降低。     

胶原生物膜在耳内镜下中耳胆脂瘤乳突根治术中的应用效果分析

摘要 目的：探讨胶原生物膜在耳内镜下乳突根治术中的应用效果。方法：选取徐州医科大学附属医院2021年4月至2022年 2月收治的51例中耳胆脂瘤患者进行回顾性分析,其中研究组27例患者予以胶原生物膜修复皮肤缺损,对照组予以颞肌筋膜修复术腔皮肤缺损,观察两组患者术后临床症状,手术时长,术腔完全上皮化时间、干耳时间及术前术后听力改变。结果：研究组术后患者因外耳道进水,存在感染及肉芽生长者1例,予以清理后未再次生长;对照组术后发生1例外耳道口狭窄的情况,予以橡胶扩张管进行扩张并后并定期清理术腔肉芽、脱落痂皮,患者外耳道恢复良好。两组术前耳闷、耳痛、耳鸣及术后耳痛VAS评分无明显差异（P>0.05）;研究组术后耳闷及耳鸣VAS评分较对照组降低（P<0.05）。研究组平均手术时长、术后术腔完全上皮化时间及平均干耳时间短于对照组（P<0.05）。两组术前术后气骨导差（ABG）、平均气导听阈（AC）比较差异均无统计学意义（P＞0.05）。结论：作为术区移植物,胶原生物膜应用于耳内镜下中耳胆脂瘤乳突根治术可加快创面术腔的修复,减少局部创伤与操作步骤,改善临床症状,缩短手术时间、术后术腔完全上皮化时间及获得干耳时间,可作为临床上有效的修复材料。     

Repair effect of thymine radical anion by echinocoside using pulse radiolysis

Repair activities of thymine radical anion by echinocoside, isolated from Pedicularis plicata. were studied using pulse radiolysis technique. The thymine radical anion was produced by the reaction of hydrated electron with thymine. Echinocoside. one of the polyphenols of phenylpropanoid glycoside, was added to the thymine aqueous solution saturated with N2. Kinetic analysis by transient absorption spectrum showed that thymine radical anion was formed at first, and then after several decades of microseconds of pulse radiolysis. the spectrum of thymine radical anion was changed to that of echinocoside radical anion. The evidence indicated that thymine radical anion was repaired through one-electron-transfer between the DNA base radical anion and echinocoside. The rate constant of electron transfer by echinocoside was 1.45× 109 dm3 · mol1 · s 1.