Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82):
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Gießen
DVG-Service
2011
|
| Ausgabe: | 1. Aufl. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | V, 207 S. Ill., graph. Darst. 21 cm |
| ISBN: | 9783863450373 |
Internformat
MARC
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| 100 | 1 | |a Sayed-Ahmed, Mohamed Zakaria |e Verfasser |0 (DE-588)1015479391 |4 aut | |
| 245 | 1 | 0 | |a Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) |c submitted by Mohamed Zakaria Sayed-Ahmed |
| 250 | |a 1. Aufl. | ||
| 264 | 1 | |a Gießen |b DVG-Service |c 2011 | |
| 300 | |a V, 207 S. |b Ill., graph. Darst. |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Zugl.: Hannover, Tierärztl. Hochsch., Diss., 2011 | ||
| 650 | 7 | |a paramyxovirus |2 cabt | |
| 650 | 7 | |a dogs |2 cabt | |
| 650 | 7 | |a sarcoma |2 cabt | |
| 650 | 7 | |a neoplasms |2 cabt | |
| 650 | 7 | |a viral deseases |2 cabt | |
| 650 | 7 | |a histopathology |2 cabt | |
| 650 | 7 | |a cell lines |2 cabt | |
| 650 | 7 | |a canine distemper virus |2 cabt | |
| 650 | 7 | |a canine parainfluenza virus |2 cabt | |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u http://d-nb.info/1015409547/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024513643&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-024513643 | |
Datensatz im Suchindex
| _version_ | 1826212231625310208 |
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| adam_text |
CONTENTS
1 INTRODUCTION 1
2 LITERATURE REVIEW 4
2.1 CANINE DISTEMPER VIRUS 4
2.1.1 OVERVIEW 4
2.1.2 CDV PROPERTIES 6
2.1.3 CANINE DISTEMPER VIRUS STRAINS R252 AND ONDERSTEPOORT 8
2.2 CANINE PARAINFLUENZA VIRUS 9
2.3 CANINE HISTIOCYIC DISORDERS 10
2.3.1 CANINE CUTANEOUS HISTIOCYTOMA 10
2.3.2 HISTIOCYTIC SARCOMA COMPLEX 11
2.3.2.1 LOCALIZED HISTIOCYTIC SARCOMA 11
2.3.2.2 DISSIMINATED HISTOCYTIC SARCOMA 12
2.3.3 CANINE REACTIVE HISTIOCYTOSIS 13
2.4 DH82 CELLS 14
2.5 VIRAL ONCOLYSIS 15
2.5.1 MECHANISMS OF ANTITUMOR EFFECTS 18
2.5.2 MECHANISMS OF ANTITUMOR SPECIFICITY 19
2.5.3 IMMUNE RESPONSE 21
2.6 CORTACTIN 21
2.6.1 DOMAIN STRUCTURE AND BINDING PARTNERS OF CORTACTIN 22
2.6.2 CORTACTIN PROMOTES CELL MIGRATION 25
2.6.3 ROLE OF CORTACTIN IN INVADOPODIA 28
2.6.4 CORTACTIN FUNCTION IN TUMORS 30
3 MATERIALS UND METHODS 32
3.1 MEDIA AND SUPPLEMENTS 32
3.2 CELL LINES 32
3.2.1 VERO CELLS 32
3.2.1.1
PROTOCOL FOR PASSAGE OF VERO CELLS 33
3.2.2 DH82 AND DH82-OND CELLS 34
3.2.2.1
PROTOCOL FOR DH82 AND DH82-OND CELL PASSAGE 34
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/1015409547
DIGITALISIERT DURCH
CONTENTS
3.3 VIRUSSTRAINS 35
3.3.1 VIRUS HARVEST 36
3.3.2 VIRUS TITRATION 36
3.3.3 INFECTION OF CELLS 37
3.4 EXPERIMENTAL DESIGN 38
3.5 HEMATOXYLIN-EOSIN (H&E)-STAINING OF CELLS 39
3.6 CELL PROLIFERATION ASSAY 40
3.6.1 CELL DOUBLING ASSAY 40
3.6.2 5-BROMO-2'-DEOXY-URIDINE-ASSAY (BRDU ASSAY) 40
3.7 IMMUNOFLUORESCENCE STAINING AND USED ANTIBODIES AND SERA 41
3.7.1 IMMUNOFLUORESCENCE FOR CDV AND CPIV 43
3.7.2 IMMUNOFLUORESCENCE FOR CORTACTIN 44
3.8 CONFOCAL MICROSCOPIC EVALUATION 44
3.9 PHAGOCYTOSIS ASSAY 44
3.10 ELECTRON MICROSCOPY 45
3.10.1
SCANNING ELECTRON MICROSCOPY (SEM) 45
3.10.2
TRANSMISSION ELECTRON MICROSCOPY (TEM) 46
3.10.2.1
EMBEDDING 46
3.10.2.2
PROCESSING OF EPON BLOCKS FOR ELECTRON MICROSCOPY 47
3.10.3
IMMUNOELECTRON MICROSCOPY 47
3.10.3.1
EMBEDDING 48
3.10.3.2
ANALYSIS OF ELECTRON MICROSCOPIC SAMPLES 49
3.11 MOLECULAR ANALYSIS 50
3.11.1
RNA ISOLATION FROM CELL CULTURES USING TRIZOL 50
3.11.2
RNA PURIFICATION UND DNASE TREATMENT USING RNEASY MINI KIT (QIAGEN) 51
3.11.3
REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION (RT-PCR) 52
3.11.4
PRIMER DESIGN 52
3.11.5
QUALITATIVE PCR FOR CREATION OF STANDARD CURVES 53
3.11.6
QUANTITATIVE PCR (RT-QPCR) 53
3.11.7
QUANTIFICATION OF DATA 55
3.12 STATISTICAL ANALYSIS 55
CONTENTS
4 RESULTS 56
4.1 MORPHOLOGICAL DESCRIPTION OF THE CELLS 56
4.1.1 VERO CELLS 56
4.1.1.1 NON-INFECTED VERO CELLS 56
4.1.1.1.1 MORPHOLOGY OF VERO CELL CULTURES 56
4.1.1.1.2 CELLULAR MORPHOLOGY OF NON INFECTED AND CDV AND CPIV INFECTED
VERO
CELLS IN HEMATOXYLIN AND EOSIN (H&E) STAINED SLIDES 56
4.1.1.1.3 GROWTH BEHAVIOR OF NON-INFECTED VERO CELLS
1
TO 10 DAYS AFTER
SEEDING 56
4.1.1.2 CPIV INFECTION OF VERO CELLS 59
4.1.1.3 CDV-OND INFECTION OF VERO CELLS 60
4.1.1.4 CDV-R252 INFECTION OF VERO CELLS 61
4.1.1.5 CDV-ONDEGFP INFECTION OF VERO CELLS 62
4.1.1.6 COMPARSION OF IMMUNOFLUORESCENCE FINDINGS OF VERO CELLS INFECTED
WITH
CPIV, CDV-OND, CDV-R252 AND CDV-ONDEGFP AT DIFFERENT TIME POINTS
POST INFECTION REGARDING TO PERCENTAGE OF INFECTED CELLS 72
4.1.2 DH82 CELLS 74
4.1.2.1 NON-INFECTED DH82 CELL CULTURE 74
4.1.2.1.1 MORPHOLOGY OF
DH82
CELL CULTURE 74
4.1.2.1.2 CELLULAR MORPHOLOGY OF NON INFECTED AND CDV AND CPIV INFECTED
DH82
CELLS IN HEMATOXYLIN AND EOSIN (H&E) STAINED SLIDES 74
4.1.2.1.3 GROWTH BEHAVIOR OF NON-INFECTED DH82 CELLS
1
TO 10 DAYS AFTER
SEEDING 75
4.1.2.2 EFFECTS OF INFECTION ON THE PROLIFERATION RATE 76
4.1.2.2.1 PROLIFERATION ASSAY 76
4.1.2.2.1.1 CELL DOUBLING 76
4.1.2.2.1.2 BRDU-ASSAY 77
4.1.2.3 INFECTION RATE ON 1,3,5 AND 10 DPI 79
4.1.2.3.1 INFECTION WITH CPIV 79
4.1.2.3.2 INFECTION WITH CDV-OND 79
4.1.2.3.3 INFECTION WITH CDV-R252 80
III
CONTENTS
4.1.2.3.4 INFECTION WITH CDV-ONDEGFP 81
4.1.2.3.5 COMPARSION OF THE INFECTION RATE OF DH82 CELLS INFECTED
WITH CPIV, AND DIFFERENT CDV STRAIN 92
4.1.3 PERSISTENTLY CDV-OND INFECTED DH82 CELLS 93
4.1.3.1 IMMUNOFLUORESCENCE OF PERSISTENTLY CDV-OND INFECTED DH82 CELLS
93
4.1.3.2 CORTACTIN EXPRESSION AND DISTRIBUTION IN NON-INFECTED AND
PERSISTENTLY CDV
OND
INFECTED DH82 CELLS 98
4.1.3.3 REVERSE TRANSCRIPTION-QUANTITATIVE POLYMERASE CHAIN REACTION
(RT-QPCR) 103
4.2 CORTACTIN EXPRESSION AND DISTRIBUTION IN NON-INFECTED, CPIV,
CDV-OND, AND
CDV-R252 INFECTED DH82 AT DIFFERENT TIME POINTS POST INFECTION 104
4.2.1 CORRELATION BETWEEN VIRUS ANTIGEN CONTAINING DH82 CELLS AND
CORTACTIN
EXPRESSION ILL
4.3 ELECTRON MICROSCOPICAL AND IMMUNO-ELECTRON MICROSCOPICAL
INVESTIGATIONS 112
4.3.1 MORPHOLOGY 112
4.3.2 ANALYSIS OF PHAGOCYTIC ACTIVITY 113
4.3.3 ANALYSIS OF THE NUMBER OF CYTOPLASMIC PROJECTIONS 114
4.3.4 ANALYSIS OF THE LENGTH/WIDTH (1/W) RATIO OF CYTOPLASMIC
PROJECTIONS 117
4.3.5 CORRELATION BETWEEN VIRUS POSITIVE DH82 CELLS AND THE NUMBER OF
CYTOPLASMIC PROCESSES 119
4.3.6 CORRELATION BETWEEN VIRUS POSITIVE DH82 CELLS AND THE MEAN (1/W)
RATIO OF
CYTOPLASMIC PROCESSES 120
4.3.7 ANALYSIS OF CORTACTIN DISTRIBUTION 121
DISCUSSION 123
5.1 INFLUENCE OF CDV-OND AND CDV-R252 INFECTION ON VERO CELLS 123
5.2 INFLUENCE OF CDV-ONDEGFP INFECTION ON VERO CELLS 126
5.3 INFLUENCE OF CANINE PARAINFLUENZA VIRUS INFECTION ON VERO CELLS 127
5.4 INFLUENCE OF CDV-OND AND CDV-R252 INFECTION ON DH82 CELLS 128
5.5 INFLUENCE OF CDV-ONDEGFP INFECTION ON DH82 CELLS 130
5.6 EFFECT OF PERSISTENT INFECTION WITH CDV-OND ON DH82 CELLS 131
5.7 INFLUENCE OF CANINE PARAINFLUENZA VIRUS INFECTION ON DH82 CELLS 131
RV
CONTENTS
5.8 EFFECTS OF INFECTION ON PHAGOCYTIC CAPACITY OF DH82 CELLS 132
5.9 EFFECTS OF PERSISTENT CDV-OND INFECTION OF DH82 CELLS ON CORTACTIN
EXPRESSION
AND DISTRIBUTION 132
5.10 EFFECTS OF
CDV-
AND CPIV INFECTION OF DH82 CELLS ON CORTACTIN EXPRESSION AND
DISTRIBUTION 133
5.11 ULTRASTRUCTURAL ANALYSIS OF CYTOPLASMIC PROCESSES IN NON-INFECTED
AND
PERSISTENTLY CDV-OND-INFECTED DH82 CELLS 134
5.12 ULTRASTRUCTURAL ANALYSIS OF CYTOPLASMIC PROCESSES IN NON-INFECTED
AND CDV
AND
CPIV INFECTED DH82 CELLS 135
5.13 CONCLUSION 135
6 SUMMARY 137
7 ZUSAMMENFASSUNG 139
8 REFERANCES 141
9 ANNEX 176
9.1 SOURCES OF CHEMICALS, REAGENTS AND ANTIBODIES 176
9.2 SOURCES FOR EQUIPMENT AND DISPOSABLE ITEMS 180
9.3 SOLUTIONS AND BUFFERS 183
9.3.1 SOLUTIONS FOR CELL CULTURE 183
9.3.2 IMMUNOFLUORESCENCE 184
9.3.2.1
PARAFORMALDEHYDE FOR CELL FIXATION 184
9.3.2.2
NUCLEAR COUNTERSTAINING WITH BISBENZIMIDE 184
9.3.3 ELECTRON MICROSCOPY 184
9.3.3.1
CACODYLATE PUFFER 0.1 M,PH 7.2 184
9.3.3.2
OSMIUM TETROXIDE (OSO4)
1 %
185
9.3.3.3
EPON 185
9.3.3.4
GLUTARDIALDEHYDE SOLUTION 2.5% 185
9.3.4 MOLECULAR BIOLOGY 185
9.4 TABLES 187
9.5 ABBREVIATION 202
10 ACKNOWLEDGEMENTS 205 |
| any_adam_object | 1 |
| author | Sayed-Ahmed, Mohamed Zakaria |
| author_GND | (DE-588)1015479391 |
| author_facet | Sayed-Ahmed, Mohamed Zakaria |
| author_role | aut |
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| dewey-full | 636.7089691801 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 636 - Animal husbandry |
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| dewey-tens | 630 - Agriculture and related technologies |
| discipline | Agrar-/Forst-/Ernährungs-/Haushaltswissenschaft / Gartenbau Medizin |
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| indexdate | 2025-03-10T13:01:39Z |
| institution | BVB |
| isbn | 9783863450373 |
| language | English |
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| physical | V, 207 S. Ill., graph. Darst. 21 cm |
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| spelling | Sayed-Ahmed, Mohamed Zakaria Verfasser (DE-588)1015479391 aut Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) submitted by Mohamed Zakaria Sayed-Ahmed 1. Aufl. Gießen DVG-Service 2011 V, 207 S. Ill., graph. Darst. 21 cm txt rdacontent n rdamedia nc rdacarrier Zugl.: Hannover, Tierärztl. Hochsch., Diss., 2011 paramyxovirus cabt dogs cabt sarcoma cabt neoplasms cabt viral deseases cabt histopathology cabt cell lines cabt canine distemper virus cabt canine parainfluenza virus cabt (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1015409547/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024513643&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Sayed-Ahmed, Mohamed Zakaria Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) paramyxovirus cabt dogs cabt sarcoma cabt neoplasms cabt viral deseases cabt histopathology cabt cell lines cabt canine distemper virus cabt canine parainfluenza virus cabt |
| subject_GND | (DE-588)4113937-9 |
| title | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) |
| title_auth | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) |
| title_exact_search | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) |
| title_full | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) submitted by Mohamed Zakaria Sayed-Ahmed |
| title_fullStr | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) submitted by Mohamed Zakaria Sayed-Ahmed |
| title_full_unstemmed | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) submitted by Mohamed Zakaria Sayed-Ahmed |
| title_short | Influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells (DH82) |
| title_sort | influence of paramyxovirus infection on phenotypical and functional characteristics of canine histiocytic sarcoma cells dh82 |
| topic | paramyxovirus cabt dogs cabt sarcoma cabt neoplasms cabt viral deseases cabt histopathology cabt cell lines cabt canine distemper virus cabt canine parainfluenza virus cabt |
| topic_facet | paramyxovirus dogs sarcoma neoplasms viral deseases histopathology cell lines canine distemper virus canine parainfluenza virus Hochschulschrift |
| url | http://d-nb.info/1015409547/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024513643&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT sayedahmedmohamedzakaria influenceofparamyxovirusinfectiononphenotypicalandfunctionalcharacteristicsofcaninehistiocyticsarcomacellsdh82 |
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