H

HD, see Huntington's disease Heat shock protein 70, transgenic mouse studies of myocardial ischemia-reperfusion injury rationale for transgenesis, 354 Southern blot analysis, 357, 359 transgenic mouse generation, 355, 357 Heme oxygenase cellular defense functions, 375-376 cytoprotection, 164 isoforms, 163-164, 351, 374 knockout mouse studies of myocardial ischemia-reperfusion injury heterozygote generation and characterization, 353-354 rationale, 351-352 targeted gene disruption, 352-353 Western blot analysis, 353 oxidative stress studies of heme oxygenase-1 activity assay biliverdin reductase preparation, 170 calculations, 171 incubation conditions, 170-171 microsomal extract preparation, 169-170 histological analysis immunohistochemical detection of enzyme and inflammatory cell markers, 173-174 inflammatory cell immunostaining, 175 MAL-2 immunostaining of oxidative damage, 175-176 tissue preparation, 172-173 lipid peroxidation measurements, 171-172

mouse models endotoxemia, 165 hypoxia exposure, 166 myocardial ischemia and reperfusion, 165-166

protein extraction from mouse tissues,

166-167 Western blot analysis, 167-169 product functions cellular effects, 374-375, 388

transgenic mouse model of neuroprotection apoptosis assays, 380-381 genotyping DNA preparation from tail, 387 polymerase chain reaction, 387-388 histochemistry brain perfusion, 376 coverslipping agents, 379 death protein staining, 377 fixatives, 378-379

hematoxylin and eosin staining, 379-380 immunostaining, 376-377 iron detection, 377-378 neuronal cell culture studies calcium measurement with fura-2,

383-384 cell culture from mouse brain, 381 chromatin condensation analysis, 382-383

glutamate induction of oxidative stress,

381-382 immunocytochemistry, 384 necrosis assay, 382

reactive oxygen species measurement, 383

reverse transcriptase-polymerase chain reaction, 385-386 RNA isolation and quantification, 385 RNA probe labeling, 386-387 viability assay, 382 Western blotting, 385 overview, 375-376 Hemoglobin iron in small molecule transfer, 187 optically transparent thin-layer electrode studies advantages, 192 allosteric effector studies, 208 cell construction, 191-192 detection systems, 194

electrode materials, 190-191 mediator use and criteria, 193-194 Nernst plots and data analysis, 204-205,

207-208 non-Nernstian response, 205-206 oxygenation relationship with redox properties, 206-207 redox potential tuning, 188-190 High-density oligonucleotide array, see Gene chip HLF, see Hypoxia-inducible factor-like factor HNE, see /eri-4-Hydroxy-2-nonenal HO, see Heme oxygenase HSP70, see Heat shock protein 70 Huntington's disease animal models, 365-366 huntingtin transgenic mouse model animal handling, 367 behavioral analysis and phenotype,

368-370 overview, 365-367, 373 oxidative damage assessment lipid peroxidation, 370-371 nitric oxide synthase activity assay, 371-373

polymerase chain reaction detection of transgene, 367-368 oxidative stress role, 366-367 Hydrogen peroxide assay in brain, 394-395 neurotoxicity, 396-397 ferr-4-Hydroxy-2-nonenal deoxyguanosine adducts abundance, 525, 534 applications of assay, 535-536 contamination precautions, 535 DNA hydrolysis, 531-532 DNA isolation from tissues homogenization, 530 protein removal, 530-531 RNase treatment, 531 high-performance liquid chromatography systems, 534-535 nuclease PI treatment, 531 phosphorous-32 postlabeling and high-performance liquid chromatography, 525-526, 528, 532-533

polynucleotide kinase treatment, 533-534 quantitative analysis and controls, 534

ring opening reduction with sodium borohydride, 533 solid-phase extraction, 532 structure, 523-524 synthesis of standards, 529-530 lipid peroxidation product, 523 protein reactivity and pathology, 523-524 Hypoxia-inducible factor-like factor, Ref-1 interactions in mammalian two-hybrid system cysteine reduction by Ref-1,7-8 false-positives, 8-10 interpretation, 7 plasmids, 4, 6 principles, 4 transfection, 6-7

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