of 3 /3
Cytoenzymology and Isozymes of Cultured Cells' 75 Subject Index Aging 18,23 Antibiotics 18, 23 Ascites tumors 26 Autoradiography 24 Autosomal trisomy 52 Bacterial contaminants 22 Blood and marrow cells and cultures I, 2, 3, 12-14, 26,27,53 Carcinogens 23 Carcinolytic agents 22 Cell line characterization 2, 16, 18,44,45 Cell line identification 44 Cell lines examined: Chang liver, Detroit 6, Detroit 6 Clone 12, HeLa, HeLa Clone 229 2-4, 12,27, 56, 57 Changes of enzyme patterns: see enzyme changes and isozyme changes Cold acetone 4, I I - 13, 54 Comparison of II tetrazolium salts 3, II, 12, 57 Contamination by other cell lines 2, 19,44 Cryogenic modalities 3, 4, 12, 13, 14, 15, 16, 56, 57 - -, cryoprotectives 3, 4,12,13,56, 57 - -, effect on histochemical enzyme reactions 12, 15, 56, 57 - -, frozen seed stock, 3, 4,56,57 - -, rapidly frozen cultures 3, 4, 56, 57 - -, slowly frozen cultures 3, 4, 56, 57 - -, storage at -85 0 C for 2 years 3, 4, 12, 16, 56, 57 Cytoenzymology 1-27, 53-57 -, in cell biology 2, 16-27 -, incubating solutions 4-7 -, of free living cells I I -, results in5 cells lines 7-II, 56, 57 Cytotoxic plastics 18 Determinants of isozyme patterns 2, 47-49 Diffusion and leaching I, 4, 7, II, 13, 14, 17 Discussion II-I7, 38-43 -, cytoenzymology I1- 17 -, isozymes 38-43 Down's disease 52 Drug toxicity evaluation 23 Electron microscopy 22,24,25,27 Electrophoresis 28 Endocrinology 24 Enzyme changes during culture 2,17-22,41-49 - -, from environmental factors 2,47 - -, from technical factors 1,12,13,17,32 -, deletions 18 -, induction 17-22, 41-49 -, pattern stability 17-22, 41-49 Enzyme, reaction products I, 7, 9, 10, 12-15,22 - - -, in supernatant 20, 21 -, variability 17-22, 41-49 Enzymes: cytoenzymology 1-27 - -, acetylcholinesterase 25 - -, acid hydrolases 22,25 - -, acid phosphatase 5, 8,9, 18,21-27 - -, adenosine triphosphatase (ATPase) 5, 8, 20, 22, 25 - -, ADPase 21,22 - -, alanine aminopeptidase 6, 8 - -, alkaline phosphatase 5, 8, 18,20,21,25-27 - -, alpha-glycerophosphate dehydrogenase 6, I I - -, alpha-keto-glutaric dehydrogenase 6, I I - -, aminopeptidases 6,8,9,18,21 - -, AMPase 20,21 - -, aryl sulfatase 6, I I - -, ATPase 5, 8, 20, 22, 25 - - beta-glucuronidase 6, 8, 24 - -, beta-hydroxybutyric dehydrogenase 6, II, 23 - -, catalase 18 - -, cathepsin 24 - -, cholinesterase 5, 8,26 - -, cytochrome c oxidase 7, 10, I I, 18-21,26 - -, delta-5-3-beta hydroxysteroid dehydrogenase24 - -, DOPA reaction 25 - -, DPN (NAD)-dependent dehydrogenases 6, 10, II - -, DPNH (NADH) diaphorase 7, 11,21-23,25 - -, esterase 5, 7, 9, 18, 27 - -, ethanol dehydrogenase 6, I I - -, glucose-6-phosphatase 6, 8, 19,20,22,26 - -, glucose-6-phosphate dehydrogenase 19, 20, 22, 26 - -, glutamic dehydrogenase 6, 10, I I - -, hydrolases 5-II, 21, 24 - -, isocitric dehydrogenase 6, I I - -, lactic dehydrogenase 6, II, 21, 24, 25 - -, leucine aminopeptidase 6, 8, 9 - -, lipase 8, 26 - -, malic dehydrogenase 6,10, II, 23 - -, 5'-nucleotidase 6,8 - -, nucleoside diphosphatase 22 - -, oxalacetic dehydrogenase 6, I I - -, oxidative enzymes 5-II, 20, 21, 23-25 - -, peroxidase 22 - -, 6-phosphogluconate dehydrogenase 23 - -, phosphamidase 26 - -, pyruvic dehydrogenase 6, I I - -, steroid dehydrogenase 24 - -, succinic dehydrogenase 6, 10, II, 19-21, 23-27 - -, thiamine pyrophosphatase 18, 22 - -, TPN (NADP)-dependent dehydrogenases 6, 10, II - -, TPNH (NADPH) diaphorase 7, 11,23,25 75

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Cytoenzymology and Isozymes of Cultured Cells' 75

Subject Index

Aging 18,23 Antibiotics 18, 23 Ascites tumors 26 Autoradiography 24 Autosomal trisomy 52

Bacterial contaminants 22 Blood and marrow cells and cultures I, 2, 3, 12-14,

26,27,53

Carcinogens 23 Carcinolytic agents 22 Cell line characterization 2, 16, 18,44,45 Cell line identification 44 Cell lines examined: Chang liver, Detroit 6, Detroit 6

Clone 12, HeLa, HeLa Clone 229 2-4, 12,27, 56, 57 Changes of enzyme patterns: see enzyme changes and

isozyme changes Cold acetone 4, I I - 13, 54 Comparison of II tetrazolium salts 3, II, 12, 57 Contamination by other cell lines 2, 19,44 Cryogenic modalities 3, 4, 12, 13, 14, 15, 16, 56, 57 - -, cryoprotectives 3, 4,12,13,56, 57 - -, effect on histochemical enzyme reactions 12, 15,

56, 57 - -, frozen seed stock, 3, 4,56,57 - -, rapidly frozen cultures 3, 4, 56, 57 - -, slowly frozen cultures 3, 4, 56, 57 - -, storage at -850 C for 2 years 3, 4, 12, 16, 56, 57 Cytoenzymology 1-27, 53-57 -, in cell biology 2, 16-27 -, incubating solutions 4-7 -, of free living cells I I -, results in5 cells lines 7-II, 56, 57 Cytotoxic plastics 18

Determinants of isozyme patterns 2, 47-49 Diffusion and leaching I, 4, 7, II, 13, 14, 17 Discussion II-I7, 38-43 -, cytoenzymology I1-17 -, isozymes 38-43 Down's disease 52 Drug toxicity evaluation 23

Electron microscopy 22,24,25,27 Electrophoresis 28 Endocrinology 24 Enzyme changes during culture 2,17-22,41-49 - -, from environmental factors 2,47 - -, from technical factors 1,12,13,17,32 -, deletions 18 -, induction 17-22, 41-49 -, pattern stability 17-22, 41-49

Enzyme, reaction products I, 7, 9, 10, 12-15,22 - - -, in supernatant 20, 21 -, variability 17-22, 41-49 Enzymes: cytoenzymology 1-27 - -, acetylcholinesterase 25 - -, acid hydrolases 22,25 - -, acid phosphatase 5, 8,9, 18,21-27 - -, adenosine triphosphatase (ATPase) 5, 8, 20, 22,

25 - -, ADPase 21,22 - -, alanine aminopeptidase 6, 8 - -, alkaline phosphatase 5, 8, 18,20,21,25-27 - -, alpha-glycerophosphate dehydrogenase 6, I I

- -, alpha-keto-glutaric dehydrogenase 6, I I - -, aminopeptidases 6,8,9,18,21 - -, AMPase 20,21 - -, aryl sulfatase 6, I I - -, ATPase 5, 8, 20, 22, 25 - - beta-glucuronidase 6, 8, 24 - -, beta-hydroxybutyric dehydrogenase 6, II, 23 - -, catalase 18 - -, cathepsin 24 - -, cholinesterase 5, 8,26 - -, cytochrome c oxidase 7, 10, I I, 18-21,26 - -, delta-5-3-beta hydroxysteroid dehydrogenase24 - -, DOPA reaction 25 - -, DPN (NAD)-dependent dehydrogenases 6, 10,

II - -, DPNH (NADH) diaphorase 7, 11,21-23,25 - -, esterase 5, 7, 9, 18, 27 - -, ethanol dehydrogenase 6, I I - -, glucose-6-phosphatase 6, 8, 19,20,22,26 - -, glucose-6-phosphate dehydrogenase 19, 20, 22,

26 - -, glutamic dehydrogenase 6, 10, I I - -, hydrolases 5-II, 21, 24 - -, isocitric dehydrogenase 6, I I - -, lactic dehydrogenase 6, II, 21, 24, 25 - -, leucine aminopeptidase 6, 8, 9 - -, lipase 8, 26 - -, malic dehydrogenase 6,10, II, 23 - -, 5'-nucleotidase 6,8 - -, nucleoside diphosphatase 22 - -, oxalacetic dehydrogenase 6, I I - -, oxidative enzymes 5-II, 20, 21, 23-25 - -, peroxidase 22 - -, 6-phosphogluconate dehydrogenase 23 - -, phosphamidase 26 - -, pyruvic dehydrogenase 6, I I - -, steroid dehydrogenase 24 - -, succinic dehydrogenase 6, 10, II, 19-21, 23-27 - -, thiamine pyrophosphatase 18, 22 - -, TPN (NADP)-dependent dehydrogenases 6, 10,

II

- -, TPNH (NADPH) diaphorase 7, 11,23,25

75

76 . P.J.~ELNITCK

Enzymes; isozymes 27-55 - -, acetyl transferase 52 - -, acid phosphatase 30, 31, 33,52 - -, adenylate kinase 52 - -, alcohol dehydrogenase 52 - -, aldolase 5 I - -, alkaline phosphatase 30,3 1,33, 52 - -, alanine aminopeptidase 30,31,33 - -, aminopeptidases 5 I - -, beta-glucuronidase 30,31,34 - -, catalase 52 - -, cholinesterase 30,31,33,51 - -, cytochrome c oxidase 37 - -, DPNH (NADH) diaphorase 38 - -, enolase 52 - -, esterase 30-32 - -, ethanol dehydrogenase 35 - -, glucose-6-phosphate dehydrogenase 34, 49, 51 - -, glutamic dehydrogenase 37 - -, hexokinase 50 - -, hydrolases 3 I - -, isocitric dehydrogenase 36,51 - -, lactic dehydrogenase 35, 5 I - -, L-amino acid oxidase 52 - -, leucine aminopeptidase 30, 34 - -, malic dehydrogenase 36,51 - -, oxidative enzymes 3 I - -, phosphoglucomutase 51 - -, pyruvic dehydrogenase 34 - -, succinic dehydrogenase 36 - -, TPNH (NADPH) diaphorase 38 - -, tryptophan synthetase 52 - -, UDPG-4-epimerase 51

Free living cells I, 4, 11-13 Freezing injury 15, 22 Frozen seed stock 3,56,57

Golgi enzymes 22

Historical data: cytoenzymology I, 17 - -, isozymes 1,2,43 Hybrid cell lines 53 Hydrogen bonding adjuvants: miscellaneous 15 - - -,PVA4 - - -,PVP4, 13, 15

Illustrations 9, 10, 12, 14, 32-37,41,43 Immobilization of enzymes and reaction products 13,

17 Immunologic technics 2, 19 Incubating solutions: cytoenzymology 4-7 - -, isozymes 29-32 In vitro conditions 2 Isozymes 1-3, 17, 19,27-55 -, changes during culture 2, 45 -, changes from environmental factors 47 -, changes from technical factors 45

Isozymes, conformational variants 47-49 -, effects of cryogenic modalities 40,41 -, effects of pathologic states 42, 43 -, in cell biology 41-43,45-50 -, incubating solutions 29-32 -, nomenclature 33,38 -, results in 5 cell lines 32-38 -, variability 41-49

Karyotyping 2, 20, 21, 26

Latent viral infections 21 Live cultures in liquid media 3 Localization of enzyme reaction products I, 4 Lysosomal enzymes 18,21-23

Materials: see cell lines examined -, cytoenzymology 1-4, 11-17,27,56,57 -, isozymes 27-32 ~ethods: cytoenzymology 3,4,11-13,16,56,57 - -, cold acetone 4,11-13,54 - -, cryogenic modalities 3,12,16,56,57 - -, cryoprotectives 3,13, 16,40,41 - -, incubating solutions 4-7 - - - -, hydrolases 5 - - - -, oxidative enzymes 7 - -, polyvinylpyrrolidone (PVP) 3, 4, 7, II, 13 - -, preparation of slides 4, 5 ~ethods: isozymes 28-32, 40, 41 - -, electrophoresis 28, 29 - -, incubating solutions 29-3 I - - - -, hydrolases 3 I - - - -, oxidative enzymes 3 I - -, preserving gels 31 - -, photographing gels 3 I - -, protein stain for starch gels 32 ~iscellaneous cells cultures 25 ~ounting medium for cytoenzymology 7 ~uscle cell culture 25

Nerve cell cultures 24 Nomenclature of isozymes 38

Permeation of cell membranes I, 4, I I, 17 Phytohemagglutinin 23 Polyvinylpyrrolidone (PVP) 3,4,7, II, 13 - -, as cryoprotective 3 - -, as hydrogen bonding adjuvant 3, 4, II, 13 - -, as mounting medium 7 PPL022 Preparation of material 4, 5,28 - -, cytoenzymology 4, 5 - -, isozymes 28 Prolonged incubation time 4, I I, 13

Radiation and cell cultures 23 Rapidly frozen cultures 3, I I

Reduced oxygen tension 23 References 58-73 Results: cytoenzymology 7-II, 56, 57 - -, hydrolases 7-1 I, 56 - -, oxidative enzymes 10, II, 57 Results: isozymes 3 1-38 - -, hydrolases 31-33 - -,oxidativeenzymes3I-38 Review of the literature: cytoenzymology 17-27 - -, isozymes 43-53

Screening carcinolytic agents 22 Slowly frozen cultures 3, I I Storage at -85 0 C 4, II, 12, 16 Summary 53-55 Synchronized cell cultures 2

Tables: hydrolases 56 -, oxidative enzymes 57 -, tetrazolium salts 57

Cytoenzymology and Isozymes of Cultured Cells . 77

Technical problems: Cytoenzymology I, 12, 13, 17 - -, isozymes 32 Temperature variation effects 23 Tetrazolium reactions 26 Tetrazolium salts 4,6,7, II, 12,26,35,38,55,57

Variants of glucose-6-phosphate dehydrogenase 49 Variations between cells 8, 9,12,20 Variations between clones 20 Viruses: cytopathic effects 18, 20, 21 -, and cell cultures 2, 18, 20-22 -, enzyme induction 2, 20, 21 -, examined with cytoenzymology; adenovirus,

Anopheles A, Coxsackie, ECHO, Friend, herpes, infectious hepatitis, influenza, Mango, measles, Moloney, myxovirus, Newcastle, oncogenic, polio, Rauscher, respiratory syncytial, Rous, vaccinia, variola, vesicular stomatitis, West Nile 2, 20-22

-, latent infection 21

X-ray mutation 20

77