Parkin介导的线粒体自噬对1型糖尿病急性心肌梗塞早期损伤的保护作用

目的:观察线粒体自噬在急性心梗(MI)早期的变化及对1型糖尿病(DM)小鼠心肌急性缺血损伤的影响。方法:将100只健康雄性C57BL/6小鼠随机分为5组,对照+假手术组(CS组);1型糖尿病+假手术组(DS组);对照+心肌梗死组(CMI组);1型糖尿病+心肌梗死组(DMI组);1型糖尿病+心肌梗死组+Parkin腺病毒过表达组(DMIPO组),每组20只。检测和比较各组小鼠的心脏功能,心肌梗死面积,心肌细胞凋亡,自噬小体含量以及心肌组织中Parkin和LC3的表达量变化。结果:与CS组相比,CMI组自噬小体含量增多,LC3II的表达量上调,Parkin的表达量明显上调(P0.05)。与CMI组比,DMI组小鼠心功能下降加剧,心梗面积增大,心肌细胞凋亡数量明显增加(P0.05),自噬水平未见明显升高。DMIPO组较DMI组自噬水平升高,心肌梗死面积减小(P0.05),心肌细胞凋亡数量减少(P0.05),心功能改善。结论:1型糖尿病通过抑制Parkin介导的心肌线粒体自噬增加心肌急性缺血损伤易感性,上调Parkin的表达可以减轻1型糖尿病时急性缺血性心肌损伤。     

白藜芦醇甙对糖尿病小鼠心肌损伤的保护作用及机制研究

目的:探讨白藜芦醇甙对小鼠小鼠心肌损伤的影响及其可能的调控机制。方法:采用腹腔注射链脲霉素的方法建立1型糖尿病小鼠模型模型,随机将雄性C57/BL6J的正常小鼠和糖尿病小鼠分为3组(每组18只):对照组、糖尿病组、糖尿病+白藜芦醇甙组。其中糖尿病+白藜芦醇甙组给予白藜芦醇甙7.5 mg/kg/d腹腔注射治疗,对照组和糖尿病组给予同体积盐水腹腔注射。药物治疗12周后,小动物心脏超声检测小鼠的心功能;天狼星红染色观察胶原变化并进一步计算胶原容积分数(collagen volume fraction,CVF);柠檬酸合酶检测试剂盒和ATP生物荧光试剂盒分别检测检测心肌柠檬酸合酶活性和ATP含量;Western blot检测Sirt3和p-AMPK的蛋白表达情况。结果:与对照组相比,糖尿病组LVEDD和LVESD均明显增加(4.823±0.103 mm和3.701±0.121 mm,P0.05),LVEF和LVFS值均明显降低(37.121±4.298%和21.023±2.187%,P0.05),CVF显著增高(20.102±1.155%,P0.05),心肌柠檬酸合酶活性和ATP含量显著降低(5.267±0.202 nmol/g和105±7.638 U/g,P0.05),Sirt3和p-AMPK蛋白表达显著降低(P0.05);与糖尿病组相比,糖尿病+白藜芦醇甙组LVEDD和LVESD均显著降低(4.354±0.113 mm和3.056±0.130 mm,P0.05),LVFS和LVEF值均显著增加(58.435±8.143%和29.071±2.232%,P0.05),CVF显著降低(10.343±0.882%,P0.05),心肌柠檬酸合酶活性和ATP含量显著增加(6.233±0.176 nmol/g和132.7±5.774 U/g,P0.05),Sirt3和p-AMPK蛋白表达显著增加(P0.05)。结论:白藜芦醇甙可改善糖尿病小鼠心肌损伤,其机制可能与白藜芦醇甙激活Sirt3/AMPK信号通路有关。     

心房钠尿肽通过上调OPA1表达抑制心衰小鼠心肌线粒体分裂并改善心脏功能

目的:探讨心房钠尿肽(Atrial natriuretic peptide,ANP)对后负荷增加引起的心脏功能下降的保护作用及其机制。方法:选择雄性C57小鼠30只,将其随机分为假手术组(sham)、主动脉弓结扎(Transverse aortic constriction,TAC)手术组和主动脉弓结扎手术ANP干预组(TAC+ANP)。ANP通过皮下注射4周,随后超声检测心脏功能、四腔心切片观察心肌重构,电镜观察心肌线粒体的形态与数量,Western-Blot检测心肌组织中融合分裂相关分子的表达。结果:同sham组相比,TAC组射血分数(Ejection fraction,EF)降低,且左室舒张末内径(End-diastolic left ventricular internal diameter,LVIDd)、左室舒张期后壁厚度(End-diastolic left ventricular posterior wall thickness,LVPWd)、左室质量(LV mass)、心肌质量/胫骨长度(Heart weight/tibial length,HW/TL)显著增加(P0.05),线粒体面积减小伴数量增加(P0.05),且线粒体融合相关蛋白OPA1表达量下降(P0.05)。同TAC组相比,TAC+ANP组EF显著增加,且LVIDd、LV mass、HW/TL均显著下降(P0.05),线粒体面积增加伴数量减少(P0.05),且线粒体融合相关蛋白OPA1表达量上调(P0.05)。在离体培养的心肌细胞中,给予ANP处理可减轻H_2O_2诱导的OPA1表达下降,给与ANP竞争性多肽抑制剂anantin后该作用消失。结论:ANP通过上调OPA1的表达抑制线粒体分裂改善后负荷增加导致的心脏功能下降。     

激活Notch1通过促进自噬改善高温高湿条件下心肌缺血/再灌注损伤

目的:明确Notch1通路在高温高湿条件下小鼠心肌缺血/再灌注(ischemia/reperfusion,I/R)损伤中的作用及其潜在机制。方法:将成年C57小鼠随机分为(1)假手术组;(2)I/R组;(3)高温高湿组;(4)高温高湿+I/R组;(5)高温高湿+Jagged1(Notch1激动剂)+I/R组;(6)高温高湿+溶剂+I/R组。采用超声心动图检测心功能,伊文氏蓝/2,3,5-三苯基氯化四氮唑双染法检测心肌梗死面积,Western blot检测Notch1细胞内段(Notch1 intracellular domain,Notch1 ICD)、Hairy和分裂增强子(Hairy and enhancer of split,Hes1)、微管相关蛋白1轻链3(microtubule-associated protein1 light chain 3,LC3)、Beclin1和p62的蛋白表达水平。结果:与假手术组对比,I/R组心功能下降,心肌梗死面积增加,Notch1 ICD、Hes1、LC3-Ⅱ/Ⅰ、Beclin1(p62相应降低)表达升高,而高温高湿组心功能下降,心肌梗死面积增加,Notch1 ICD、Hes1、LC3-Ⅱ/Ⅰ、Beclin1(p62相应升高)表达降低;和I/R组或高温高湿组对比,高温高湿+I/R组心功能进一步下降,心肌梗死面积进一步增加,Notch1 ICD、Hes1、LC3-Ⅱ/Ⅰ、Beclin1(p62进一步升高)表达进一步降低;和高温高湿+I/R组对比,加入Notch1激动剂Jagged1后,心功能提高,心肌梗死面积缩小,Notch1 ICD、Hes1、LC3-Ⅱ/Ⅰ、Beclin1(p62相应降低)表达升高。结论:激活Notch1通路可能通过促进自噬从而缓解高温高湿所致的心肌缺血/再灌注损伤。     

富氢液对脓毒症小鼠心肌细胞线粒体自噬的影响

为评价富氢液（hydrogen-rich saline,HRS）对脓毒症小鼠心肌细胞线粒体自噬的调节及其对心功能障碍的治疗作用,选取72只雄性C57BL/6J小鼠作为研究对象,采用随机数字表法分为假手术组（Sham组）、假手术+富氢液组（Sham+HRS组）、脓毒症组（CLP组）、CLP+富氢液组（CLP+HRS组）,每组18只。采用盲肠结扎穿孔法建立小鼠CLP模型。Sham+HRS组和CLP+HRS组分别于造模后1、6 h时腹腔注射富氢液10 mL·kg^-1。每组随机取6只小鼠,于造模后24 h时收集小鼠颈动脉血样,采用ELISA法测定血液肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）、白细胞介素（interleukin-1β,IL-1β）、肌钙蛋白I（cardiac troponin I,cTnI）和肌酸激酶同工酶（creatine kinase MB,CK-MB）水平;于造模后24 h时取心肌组织,采用荧光素-荧光酶发光法检测ATP,荧光分光光度法检测线粒体膜电位（mitochondrial membrane potential,MMP）。造模后24 h采用Western blot法测定心肌组织线粒体自噬相关蛋白微管关联蛋白轻链3Ⅱ/轻链3Ⅰ（microtubule-associated protein 1 light/protein 3 light,LC3Ⅱ/LC3Ⅰ）和蛋白62（protein 62,P62）的表达水平。结果显示,与Sham组比较,CLP组血清TNF-α、IL-1β、cTnI和CK-MB水平升高,心肌ATP、MMP水平下降,心肌LC3Ⅱ/LC3Ⅰ表达水平上调,P62表达水平下调,差异有统计学意义（P<0.05）;与CLP组比较,CLP+HRS组血清TNF-α、IL-1β、cTnI和CK-MB含量下降,心肌组织ATP、MMP水平升高,LC3Ⅱ/LC3Ⅰ表达水平进一步上调,P62表达进一步下调（P<0.05）。结果表明,富氢液对脓毒症小鼠心功能障碍的治疗作用可能是通过调节心肌细胞线粒体自噬实现的。研究旨在探讨富氢液对脓毒症小鼠心功能障碍的治疗作用及机制,以期为富氢液的临床转化提供理论依据。     

激活κ-阿片受体抑制小鼠心肌缺血再灌注损伤的作用及机制

目的:探讨外源性κ-阿片受体激动剂U50,488H对小鼠缺血再灌注损伤心肌的保护作用及其机制。方法:选择成年雄性C57小鼠40只,将其随机分为4组:假手术组(Sham),缺血再灌注组(I/R),κ-阿片受体激动剂U50,488H+I/R组(U+I/R),κ-阿片受体阻断剂nor-BNI+U50,488H+I/R组(N+U+I/R)。建立小鼠急性心肌缺血再灌注在体模型,通过小动物超声仪检测小鼠心功能,采用氯化三苯基四氮唑-伊文思蓝双染检测心肌梗死面积,检测血清心肌损伤物LDH活性和cTnI含量,Western-Blot检测Ca MKII和磷酸化Ca MKII的表达。结果:与Sham组相比,I/R组小鼠心功能下降,心肌梗死面积增加,血清LDH和cTnI水平升高(P0.05),心肌组织内磷酸化Ca MKII的表达明显增加(P0.05);与I/R组相比,U+I/R组心功能改善,心肌梗死面积减小,血清LDH和cTnI水平降低(P0.05),心肌组织内CaMKII磷酸化被抑制(P0.05)。给予nor-BNI后,上述U50,488H的作用均被阻断。结论:κ-阿片受体激活可抑制CaMKII磷酸化并抑制心肌缺血再灌注损伤,改善心功能。     

白藜芦醇甙减轻高糖所致心肌微血管内皮细胞损伤的作用及机制研究

目的:探讨白藜芦醇甙在高糖处理的大鼠心肌微血管内皮细胞损伤中的作用及其可能调控机制。方法:酶消法分离大鼠CMECs,高糖处理CMECs建立细胞损伤模型,实验随机分为6个组:对照组(葡萄糖浓度为5.5 mmol/L)、白藜芦醇甙组、高糖组(葡萄糖浓度为33 mmol/L)、高糖+白藜芦醇甙组、高糖+白藜芦醇甙+3-MA(自噬抑制剂)组和高糖+雷帕霉素(自噬诱导剂)组。白藜芦醇甙组和高糖+白藜芦醇甙组分别加入10μmol/L的白藜芦醇甙孵育24 h,高糖+白藜芦醇甙+3-MA组加入10μmol/L的白藜芦醇甙和10μmmol/L 3-MA孵育24 h,高糖+雷帕霉素组加入100 nmol/L的雷帕霉素孵育24小时。CCK-8实验检测大鼠CMECs增殖;Tunel法检测大鼠CMECs凋亡;FITC-葡聚糖清除实验检测单层CMECs通透性;Western blot检测LC3Ⅱ和p62的表达。结果:与对照组和白藜芦醇甙组相比,高糖组CMECs增殖能力降低(P<0.05),凋亡率显著增加(P<0.05),细胞通透性增加(P<0.05),LC3Ⅱ表达降低(P<0.05),p62的表达增加(P<0.05);与高糖组相比,高糖+白藜芦醇甙组和高糖+雷帕霉素组CMECs增殖能力增加(P<0.05),凋亡率显著降低(P<0.05),细胞通透性降低(P<0.05),LC3Ⅱ表达增加(P<0.05),p62的表达降低(P<0.05);与高糖+白藜芦醇甙组相比,高糖+白藜芦醇甙+3-MA组CMECs增殖能力降低(P<0.05),凋亡率显著增加(P<0.05),细胞通透性增加(P<0.05),LC3Ⅱ表达降低(P<0.05),p62的表达增加(P<0.05)。结论:白藜芦醇甙通过增加自噬减轻高糖处理的大鼠心肌微血管内皮细胞损伤。     

法舒地尔对急性心肌梗死大鼠心肌组织中Rho激酶的影响及心肌保护作用(英文)

目的:分析急性心肌梗死(AMI)后大鼠心肌组织Rho激酶表达的变化及心肌细胞凋亡情况,观察法舒地尔对急性心肌梗死(AMI)后大鼠心肌组织Rho激酶表达的影响,探讨法舒地尔对心梗后心肌的保护作用。方法:选取雄性Wistar大鼠,随机分为三组:治疗组、AMI组、假手术组。治疗组及AMI组均结扎左前降支(LAD)制作AMI模型;假手术组只在其LAD下穿线不结扎。治疗组给予法舒地尔5mg/kg,腹腔注射,每日两次;对照组和假手术组给予等量生理盐水。1周后,EvensBlue及NBT双染色确定缺血面积及梗死面积,RT-PCR法测定rho激酶mRNA的表达,DNA断裂的原位末端标记法(T UNEL法)检测缺血区心肌细胞凋亡指数(AI),免疫组化测定凋亡相关蛋白bcl-2及bax表达的变化。结果:1周后,AMI组与假手术组相比,AMI组大鼠Rho激酶mRNA表达增加(P0.01),凋亡相关蛋白bax表达增加(P0.01),bcl-2表达减少(P0.01),AI明显增加(P0.01)。治疗组与AMI组相比,梗死面积显著减小(P0.05),Rho激酶mRNA及bax表达显著减少,AI显著降低,bcl-2表达显著增加(均P0.01)。结论:大鼠AMI后,心肌组织中Rho激酶的表达增加,心肌细胞凋亡指数增加,连续应用法舒地尔1周能有效减少心肌细胞凋亡指数,起到心肌保护的作用。     

人参皂苷Rb1通过上调PGC-1α缓解糖尿病心肌病

目的:研究人参皂苷Rb1对糖尿病心肌病的治疗作用并阐明其分子机制。方法:采用腹腔注射链脲佐菌素的方法,建立糖尿病心肌病动物模型。将小鼠分为3组:WT组,DM组,DM+Rb1组。超声心动图分析小鼠心功能;Western blot分析PGC-1α、cleaved caspase-3、bcl-2等蛋白表达;MitoSOX染色分析线粒体ROS含量;透射电镜分析线粒体数目。结果:与WT组相比,DM组小鼠心功能显著下降(LVEF,P<0.01),PGC-1α表达下调(P<0.01),线粒体数目减少(P<0.01);而Rb1处理后,显著改善了DM小鼠心功能(LVEF,P<0.01),恢复了PGC-1α表达(P<0.05),增加了线粒体数目(P<0.05)。同时,Rb1处理后,减少了糖尿病小鼠心肌线粒体ROS产生(P<0.01),恢复了bcl-2蛋白表达(P<0.01),降低了cleaved caspase-3蛋白表达(P<0.01),从而减少了高糖引起的细胞凋亡(P<0.05)。而siPGC-1α处理后,阻断了Rb1的上述作用。结论:人参皂苷Rb1通过上调PGC-1α改善糖尿病小鼠心功能,缓解糖尿病心肌病。其机制可能与人参皂苷Rb1降低心肌线粒体ROS产生并减少心肌细胞凋亡有关。     

双氢杨梅素对缺氧诱导的心肌细胞炎症反应和凋亡的影响及机制

目的:探讨双氢杨梅素对缺氧诱导的心肌细胞炎症和凋亡的影响及其可能调控机制。方法:将心肌细胞(H9C2细胞系)缺氧处理建立细胞模型,随机分为4组:对照组、对照+双氢杨梅素组、缺氧组、缺氧+双氢杨梅素组。对照+双氢杨梅素组和缺氧+双氢杨梅素组分别加入5μmol/L的双氢杨梅素孵育24 h,对照组和缺氧组分别加入同等体积的DMSO孵育24 h。实时PCR检测心肌细胞CD40、IL-1β、IL-6、TNF-α、mi RNA-145-5p的m RNA水平;ELISA检测IL-1β、IL-6和TNF-α的浓度;MTT实验检测心肌细胞生存率;Tunel法检测心肌细胞凋亡;Western blot检测心肌细胞Cleaved Caspase3的表达。结果:与对照组和对照+双氢杨梅素组相比,缺氧组CD40、IL-1β、IL-6、TNF-α的m RNA水平升高(P0.05),IL-1β、IL-6、TNF-α浓度显著升高(P0.05),mi RNA-145-5p的m RNA水平显著降低(P0.05),心肌细胞生存率显著降低(P0.05),心肌细胞凋亡显著增加(P0.05),Cleaved Caspase3的蛋白表达显著增加;与缺氧组相比,缺氧+双氢杨梅素组CD40、IL-1β、IL-6、TNF-α的m RNA水平显著降低(P0.05),IL-1β、IL-6、TNF-α浓度显著降低(P0.05),mi RNA-145-5p的m RNA水平显著增加(P0.05),心肌细胞生存率显著增加(P0.05),心肌细胞凋亡显著降低(P0.05),Cleaved Caspase3的蛋白表达显著降低。结论:双氢杨梅素可抑制缺氧诱导的心肌细胞炎症和凋亡,其机制可能与双氢杨梅素上调mi RNA-145-5p进一步抑制CD40介导的炎症反应和凋亡有关。     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Chemokines and cytokines: axis and allies in asthma and allergy

Allergic asthma can be precipitated by many factors. For the atopic person, fungus, pollen, dust mites, cockroach antigens, and diesel exhaust are all agents that may trigger an allergic attack. Cytokines and chemokines are integral mediators of fungal asthma. From the earliest time points, they recruit and activate the cells required for the clearance of fungus as well as being critical factors involved in the immunopathology of this disease. In the final analysis, it is clear that these mediators can act to the benefit or the detriment of the host.     

Oxidation and nitration of ribonuclease and lysozyme by peroxynitrite and myeloperoxidase

In spite of the many studies on protein modifications by reactive species, knowledge about the products resulting from the oxidation of protein-aromatic residues, including protein-derived radicals and their stable products, remains limited. Here, we compared the oxidative modifications promoted by peroxynitrite and myeloperoxidase/hydrogen peroxide/nitrite in two model proteins, ribonuclease (6Tyr) and lysozyme (3Tyr/6Trp). The formation of protein-derived radicals and products was higher at pH 5.4 and 7.4 for myeloperoxidase and peroxynitrite, respectively. The main product was 3-nitro-Tyr for both proteins and oxidants. Lysozyme rendered similar yields of nitro-Trp, particularly when oxidized by peroxynitrite. Hydroxylated and dimerized products of Trp and Tyr were also produced, but in lower yields. Localization of the main modified residues indicates that peroxynitrite decomposes to radicals within the proteins behaving less specifically than myeloperoxidase. Nitrogen dioxide is emphasized as an important protein modifier.