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MITOCHONDRIAL REDOX IMAGING FOR CANCER DIAGNOSTIC AND THERAPEUTIC STUDIES

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Journal of Innovative Optical Health Sciences 2009年第4期2卷 325-341页

作者： LIN Z.LI HE N.XU MAHSA RANJI SHOKO NIOKA BRITTON CHANCE Department of Radiology School of Medicine University of PennsylvaniaB6 Blockley Hall423 Guardian Drive PhiladelphiaPA 19104-6069USA The Institute for Translational Medicine and Therapeutics School of Medicine University of Pennsylvania PhiladelphiaPA 19104USA Biophotonics Laboratory Department of Electrical Engineering and Computer Science University of Wisconsin-Milwaukee MilwaukeeWI 53211USA Johnson Research Foundation Department of Biochemistry and Molecular Biophysics School of Medicine University of Pennsylvania 250 Anatomy Chemistry BuildingPhiladelphiaPA 19104USA

Mitochondrial redox states provide important information about energy-linked biological processes and signaling events in tissues for various disease phenotypes including cancer.The redox scanning method developed at the Chance laboratory about 30 years ago has allowed 3D highresolution(∼50×50×10µm^(3))imaging of mitochondrial redox state in tissue on the basis of the fluorescence of NADH(reduced nicotinamide adenine dinucleotide)and Fp(oxidized flavoproteins including flavin adenine dinucleotide,i.e.,fad).In this review,we illustrate its basic principles,recent technical developments,and biomedical applications to cancer diagnostic and therapeutic studies in small animal models.Recently developed calibration procedures for the redox imaging using reference standards allow quantification of nominal NADH and Fp concentrations,and the concentration-based redox ratios,e.g.,Fp/(Fp+NADH)and NADH/(Fp+NADH)in tissues.This calibration facilitates the comparison of redox imaging results acquired for different metabolic states at different times and/or with different instrumental settings.A redox imager using a CCD detector has been developed to acquire 3D images faster and with a higher in-plane resolution down to 10µm.Ex vivo imaging and in vivo imaging of tissue mitochondrial redox status have been demonstrated with the CCD imager.Applications of tissue redox imaging in small animal cancer models include metabolic imaging of glioma and myc-induced mouse mammary tumors,predicting the metastatic potentials of human melanoma and breast cancer mouse xenografts,differentiating precancerous and normal tissues,and monitoring the tumor treatment response to photodynamic therapy.Possible future directions for the development of redox imaging are also discussed.

关键词： Redox ratio reduced nicotinamide adenine dinucleotide NADH flavoprotein flavin adenine dinucleotide fad calibration

ISOLATION AND PROPERTIES OF L-AMINO ACID OXIDASE FROM OPHIOPHAGUS HANNAH VENOM

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Science China Chemistry 1982年第9期25卷 941-952页

作者：胡纫秋王锦兰雷克健 Institute of Biophysics Beijing Academia Sinica

This paper reports the isolation and some properties of L-amino acid oxidase from Ophi-ophagus hannah venom. The differences between L-amino acid oxidase from Ophiophagusohannah and that from other sources in specific activity, properties and the spectrum of iso-zymes are noticeable. The result of electrophoresis on polyacrylamide gel shows that this purified enzyme ishomogenous. The molecular weight determined by gradient polyacrylamide gel slab （4--30%）is around 15×10dalton. The molecular weight of the subunit determined by SDS gradient gelelectrephoresis （4--30%） is around 7.3×10dalton. Therefore, this enzyme is composed of twosubunits. The absorption spectrum reveals that there are two fads in each molecule. The optimum pH of enzymic reaction is around 8.7--9.0 when L-leucine is used as substrate.The inhibition of the preducts is noticeable when substrate concentration goes beyond 3mM.This enzyme is heat stable and its activity would not decrease obviously after heating at 55℃for 40 min. A linear relationship between enzyme concentration and reaction rate was noticedwhen enzyme concentration was below 5.7μg/ml.

关键词： ISOLATION AND PROPERTIES OF L-AMINO ACID OXIDASE FROM OPHIOPHAGUS HANNAH VENOM fad ACID

PURIFICATION AND SOME PROPERTIES OF GLYCEROL-3-PHOSPHATE DEHYDROGENASE FROM RABBIT SKELETAL MUSCLE MITOCHONDRIA

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Science in China,Ser.B 1986年第10期29卷 1027-1038页

作者：施善平刘建华林其谁 Shanghai Institute of Biochemistry Academia Sinica

Glycerol-3-phosphate dehydrogenase (E. C. 1. 1. 99. 5) was solubilized from rabbit skeletal muscle mitochondria by Triton X-100 and purified through hydroxyapatite column chromatography, DEAE-Sepharose CL-6B column chromatography and sucrose density gradient ultracentrifugation. The preparation was electrophoretically pure, the total recovery was 10% and the specific activity had been increased 200-fold. The apparent molecular weight of the enzyme polypeptide was 69,000, it existed in the form of enzyme-Triton X-100 complex with a Stokes’ radius of 59 and a sedimentation coefficient of 10.7S. There were 1.7mg Triton X-100 and 26 μg phospholipid per mg protein of the preparation. The enzyme absorbed at 410 and 460 nm which could be attributed to non-haem iron and fad respectively. Both of the absorption would be largely diminished by adding the substrate glycerol-3-phosphate.

关键词： fad PURIFICATION AND SOME PROPERTIES OF GLYCEROL-3-PHOSPHATE DEHYDROGENASE FROM RABBIT SKELETAL MUSCLE MITOCHONDRIA DEAE der Pro

Pulse radiolysis study of rapid electron transfer from electron adduct and reducing OH adduct of dAMP to riboflavin and flavin adenine dinucleotide (fad)

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Chinese Journal of Chemistry 2000年第4期18卷 500-505页

作者：陆长元韩镇辉屠铁城苗金玲姚思德林念芸 Laboratory of Radiation Chemistry Shanghai Institute of Nuclear Research Chinese Academy of Sciences Shanghai 201800 China

Rapid electron transfer from electron adduct and reducing OH adduct of 2-deoxyadenosine-5′-monophosphate acid (dAMP) to riboflavin (RF) and flavin adenine dinucleotide (fad) was studied using the time-resolved pulse radiolysis techniques. Both spectroscope and kinetic analyses showed that transient absorption spectrum of electron adduct or OH adduct of dAMP formed at first, and men changed to that of radical anion of RF or fad after several microseconds of pulse. The evidence indicated that electron transfer from electron adduct and reducing OH adduct of dAMP to RF or fad did occur. From buildup or bleaching kinetics of radical anions of RF and fad, the rate constants for electron transfer were determined, respectively.

关键词： Electron adduct of dAMP reducing OH adduct of dAMP riboflavin (RF) fad electron transfer pulse radiolysis

Proposed GSSP for the base of Cambrian Stage 9,coinciding with the first appearance of Agnostotes orientalis, at Duibian, Zhejiang, China

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Science China Earth Sciences 2009年第4期52卷 434-451页

作者： Loren E. BABCOCK Gabriella BAGNOLI School of Earth Sciences The Ohio State University Department of Earth Sciences of Pisa University

Exposed in natural outcrops near the Duibian Village, Jiangshan County, Zhejiang Province, China, the Duibian B section is proposed as the boundary stratotype for the base of an unnamed stage termed provisionally Cambrian Stage 9. The proposed position of the GSSP is 108.12 m above the base of the Huayansi Formation, at a horizon coinciding with the first appearance of the cosmopolitan agnostoid trilobite Agnostotes orientalis. This horizon coincides also with the first appearance of the cosmopoli-tan polymerid trilobite Irvingella angustilimbata. The section fulfills all the requirements for a GSSP, and the horizon can be constrained not only by the primary stratigraphic marker (A. orientalis) but also with secondary biostratigraphic, chemostratigraphic, and sequence-stratigraphic correlation tools. The first appearance of A. orientalis is one of the most readily recognizable levels in Cambrian, and can be correlated with precision to all paleocontinents.

关键词： Cambrian GSSP fad of Agnostotes orientalis China chronostratigraphy

INTRINSIC FLUORESCENCE CHANGES IN NERVE TERMINALS,EVOKED BY ACTION POTENTIALS,ARE MODULATED BY KREBS CYCLE SUBSTRATES

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Journal of Innovative Optical Health Sciences 2011年第2期4卷 209-214页

作者： PAUL KOSTERIN ANA LIA OBAID BRIAN M.SALZBERG Department of Neuroscience University of Pennsylvania School of Medicine Philadelphia PA 19104USA Department of Physiology University of Pennsylvania School of Medicine Philadelphia PA 19104USA

Electrical stimulation of the mammalian neurohypophysial infundibular stalk evokes the entry of Nat and Ca^(2+) into the neurosecretory terminals during the action potential.These events,in turn,increase intracellular Ca^(2+) and activate NaK-and Ca-ATPases,prompting the mitochondria to increase oxidative phosphorylation which can be monitored by recording the changes in fad and NADHfluorescence.This paper reflects our efforts to determine whether or not modulating the capacity of mitochondria to produce ATP,by changing the concentrations of two important substrates of the Krebs cycle of the nerve terminal mitochondria,pyruvate and glucose,has an effect on the intrinsicfluorescence changes triggered by action potential stimulation.

关键词： Fluorescence fad NADH action potential metabolism

Direct correlation of radiolarian Kilinora spiralis Zone with the Late Jurassic ammonite faunal succession in the Kurisaka Formation, Kurosegawa Terrane, SW Japan

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Science China Earth Sciences 2009年第12期52卷 1910-1923页

作者： ISHIDA Keisuke TSUJINO Yasuyuki KOZAI Takeshi SATO Tadashi HIRSCH Francis Laboratory of Geology Institute of SAS Faculty of Science University of Tokushima Tokushima 770-8502 Japan Tokushima Prefectural Museum Bunka-no-Mori Park Tokushima 770-8070 Japan Laboratory of Geosciences Naruto University of Education Naruto 772-8502 Japan Fukada Geological Institute Hon-Komagome 2-13-12 Bunkyo-Ku Tokyo 113-0021 Japan

The last appearance datum of the radiolarian Kilinora spiralis is recorded above the first appearance datum of the ammonite Ataxioceras (A.) kurisakense in the Todoro Section of the Kurisaka Formation, Southern Kurosegawa Terrane, Shikoku, SW Japan. The constraint by ammonite age prolongs the range of the Kilinora spiralis Zone, a remarkable Jurassic radiolarian zone in Japan-NW Pacific region, into the lower Kimmeridgian. The direct correlation of the Kilinora spiralis zone with the Late Jurassic ammonite faunal succession in the Kurisaka Formation will provide a clue to the still pending chronological difference between European and North American radiolarian zones.

关键词： Late Jurassic LAD of Kilinora spiralis radiolarian zonation fad of Ataxioceras (A.) kurisakense Kurisaka Formation Kurosegawa Terrane SW Japan

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多光子激发荧光与毛细管电泳联用的实验研究

光谱学与光谱分析 2005年第4期25卷 502-505页

作者：孙云旭朱峰马万云清华大学物理系原子分子纳米科学教育部重点实验室北京100084

将多光子激发荧光探测与毛细管电泳技术相结合,研制了多光子激发荧光毛细管电泳联用装置。这种方法可以高效快速的分离检测复杂样品中多种不同的荧光分子。作者对5HT ,fad ,NADH这三种重要的生物分子,不用染料标记,分别用双光子激发和... 详细信息

将多光子激发荧光探测与毛细管电泳技术相结合,研制了多光子激发荧光毛细管电泳联用装置。这种方法可以高效快速的分离检测复杂样品中多种不同的荧光分子。作者对5HT ,fad ,NADH这三种重要的生物分子,不用染料标记,分别用双光子激发和三光子激发,进行了直接的分离、识别和检测。得到的检测限分别是5HT 1. 0×10 -6mol·L-1,fad 7. 4×10 -7mol·L-1,NADH 9. 8×10 -7mol·L-1。5HT的检测限比紫外吸收低2个数量级;fad和NADH的检测限比紫外吸收低1个数量级。

关键词：激发荧光实验研究毛细管电泳技术 NADH 紫外吸收多光子激发双光子激发检测限荧光探测联用装置荧光分子复杂样品分离检测生物分子 fad 数量级

亚油酸与核黄素或fad激发态之间电荷转移的直接证据

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化学学报 2001年第11期59卷 1858-1863页

作者：陆长元刘官树韩镇辉唐红飞姚思德林念芸陈君男毛裕民中国科学院上海原子核研究所复旦大学遗传所

根据荧光显微镜方法,我们首次发现核黄素(维生素B2)主要分布在细胞核的膜上和核的内部,故核黄素光敏化与辐射敏化的靶位置主要集中在细胞核内;当核黄素的浓度较大时,细胞膜上也有药物的分布,即在高浓度时,细胞膜也是光敏化与辐射敏化的... 详细信息

根据荧光显微镜方法,我们首次发现核黄素(维生素B2)主要分布在细胞核的膜上和核的内部,故核黄素光敏化与辐射敏化的靶位置主要集中在细胞核内;当核黄素的浓度较大时,细胞膜上也有药物的分布,即在高浓度时,细胞膜也是光敏化与辐射敏化的作用位点之一.应用308nm激光光解时间分辨吸收方法,以亚油酸作为脂质的模型化合物,研究了亚油酸与核黄素和黄素腺嘌呤二核苷酸(fad)的激发三重态之间的电荷转移过程,首次给出了电荷转移的直接证据.

关键词：亚油酸核黄素 DNA 辐射损伤光敏化黄素腺嘌呤二核苷酸荧光显微镜电荷转移 fad 激发态