Molecular biological studies into the pathophysiology of osteoporosis:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | Undetermined |
| Veröffentlicht: |
1998
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | München, Techn. Univ., Habil.-Schr., 1998 |
| Beschreibung: | III, 87 Bl. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Atkinson, Michael John |e Verfasser |0 (DE-588)131634100 |4 aut | |
| 245 | 1 | 0 | |a Molecular biological studies into the pathophysiology of osteoporosis |c vorgelegt von Michael J. Atkinson |
| 264 | 1 | |c 1998 | |
| 300 | |a III, 87 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Techn. Univ., Habil.-Schr., 1998 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-008715505 | |
Datensatz im Suchindex
| _version_ | 1812461202276614144 |
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| adam_text |
1 INTRODUCTION 1
1.1. SKELETAL DYNAMICS 1
1.1.1 ANATOMY OF THE SKELETON
1
1.1.2 GROWTH 2
1.1.3 MODELLING
3
1.1.4 REMODELLING
3
1.2. THE REGULATION OF BONE TURNOVER AND OSTEOPOROSIS 3
1.2.1 THE BONE RESORBING UNIT (BRU)
3
1.2.2 METABOLIC BONE DISEASE
4
1.2.3 BONE STRENGTH AND FRACTURE
% 5
1.3. CELL BIOLOGY OF BONE 6
1.3.1 OSTEOBLASTS
6
1.3.2 OSTEOCYTES 9
1.33 OSTEOCLASTS
10
1.3.4 REGULATION OF BONE CELL ACTIVITY
10
1.3.4.1 CALCIOTROPIC ENDOCRINE SYSTEM 10
1.3.4.2 IMMUNE SYSTEM AND THE SKELETON 11
1.3.4.3 LOCAL GROWTH FACTORS 13
1.4. OSTEOPOROTIC BONE DISEASE 14
1.4.1 EPIDEMIOLOGY AND PATHOPHYSIOLOGY OF OSTEOPOROSIS
14
1.42 PRIMARY "INVOLUTIONAL" OSTEOPOROSIS
15
1.4.2.1 POST MENOPAUSAL (TYPE I) OSTEOPOROSIS 15
1.4.2.2 SENILE (TYPE II) OSTEOPOROSIS 17
1.4.2.3 IDIOPATHIC JUVENILE OSTEOPOROSIS 17
1.4.3 SECONDARY OSTEOPOROSIS
17
1.4.4 OSTEOPOROSIS RISK FACTORS
18
1.45 SOCIO-ECONOMIC CONSEQUENCES
18
1.4.6 ANTIRESORPTIVE AND ANABOLIC THERAPEUTIC STRATEGIES
19
1.4.6.1 OESTROGEN 20
1.4.6.2 BISPHOSPHONATES 20
1.4.6.3 CALCITONIN 20
1.4.6.4 FLUORIDE 21
1.4.6.5 PARATHYROID HORMONE 21
1.4.6.6 ANDROGENS 21
1.4.7 FUTURE THERAPIES
22
1.5 OBJECTIVES 22
2 METHODS AND MATERIALS 24
2.1 CELL CULTURE 24
2.1.1 CELL LINES
24
2.12 GENERAL CULTURE CONDITIONS
25
2.13 CELL VIABILITY
26
2.1.4 PLATING EFFICIENCY
26
2.15 ROS 1712.8 CELL CULTURE
26
2.1.6 IN VITRO ANALYSIS OF OSTEOBLAST DIFFERENTIATION 26
2.1.7 TRANSIENT TRANSFECTION OF COS 7 CELLS
26
2.1.8 IMMUNOSCREENING OF CELLS TRANSFECTED WITH ME A B ELL
27
2.2 ANALYSIS OF APOPTOTIC CELL DEATH 27
2.2.1 FRAGMENTATION OF GENOMIC DNA
27
2 2.2 TUNEL STAINING
27
22.3 ULTRASTRUCTURAL MORPHOLOGY
28
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/959419756
2.3 PROTEIN ANALYSIS 28
2.3.1 BIOSYNTHETIC LABELLING
28
2.32
S9FE-AND 223RA LABELLING 28
233 IMMUNOPRECIPITATION OF LABELLED PROTEINS
28
2
3.4 DENATURING POLYACRYLAMIDE GEL ELECTROPHORESIS (PAGE) 29
233 NON-DENATURING PAGE 29
2.3.6 WESTERN BLOTTING
29
2.4 MOLECULAR BIOLOGY 29
2.4.1 HYBRIDISATION PROBES
29
2.4.2 VECTORS LIST (INCLUDING E. COLI HOSTS) '
31
2.4.3 LABELLING OF PROBES
32
2.4.3.1 RADIOACTIVE LABELLING OF CDNA PROBES 32
2.4.3.2 END-LABELLING OF OLIGONUCLEOTIDE 32
2.4.3.3 DIGOXYGENIN-LABELLING OF CRNA PROBES 32
FOR IN SITU HYBRIDISATION
2.4.3.4 GENERATION OF SINGLE STRANDED CDNA PROBES FOR 33
DIFFERENTIAL HYBRIDISATION REACTIONS
2.4.4 BACTERIAL CULTURE 33
2.4.4.1 CHEMICAL TRANSFECTION _ 33
2.4.4.2 ELECTROPORATION 33
2.4.4.3 PLASMID DNA PREPARATION 33
2.43 CDNA LIBRARIES
34
2.4.5.1 ROS 17/2.8 CDNA LIBRARY IN LAMBDA GTLO 34
2.4.5.2 ROS 17/2.8 LIBRARY IN PCDNA 1 34
2.4.5.3 ROS 25/1 LIBRARY IN PCDNA 1 34
2.4.5.4 SUBTRACTIVE ROS 17/2.8 CDNA LIBRARY 34
2.4.6 RNA AND DNA ANALYSIS 35
2.4.6.1 RNA PREPARATION 35
2.4.6.2 NORTHERN BLOT 36
2.4.6.3 DNA SEQUENCING 35
2.4.6.4 PCR 35
2.4.6.5 SEQUENCE ANALYSIS 36
2.4.7 OTHER MOLECULAR BIOLOGICAL TECHNIQUES
36
2.5 SCREENING STRATEGIES 36
23.1 DIFFERENTIAL HYBRIDISATION OF LAMBDA GTLO
36
ROS 17/2.8 LIBRARY
232 SUBTRACTION HYBRIDISATION OF THE ROS 17/2.8 AND 36
ROS 25/1 PCDNA 1 LIBRARIES
233 DIFFERENTIAL SCREENING OF SUBTRACTIVE ROS 17/2.8 LIBRARY 37
2.5.4 EXPRESSION CLONING 37
2.6 STATISTICAL ANALYSIS 38
3 RESULTS 39
3.1 IDENTIFICATION OF MARKERS OF OSTEOBLAST DIFFERENTIATION 39
BY DIFFERENTIAL HYBRIDISATION SCREENING
3.1.1 THYMOSIN P 4 40
3.12 FERRITIN
42
3.1.2.1 FERRITIN PROTEIN EXPRESSION 43
3.1.2.2 METAL SEQUESTRATION BY OSTEOBLASTIC CELLS 44
3.13 OTHER DIFFERENTIALLY EXPRESSED CDNA CLONES (ESTS) 45
3.2 IDENTIFICATION OF MARKERS OF OSTEOBLAST DIFFERENTIATION 45
BY SUBTRACTIVE HYBRIDISATION
32.1 RIBOSOMAL PROTEIN MRNAS 46
322 FERRITIN LIGHT CHAIN
46
U
3.2.3 NOVEL ESTS
46
32.4 CLONE H57
47
3.2.4.1 PUTATIVE SECONDARY STRUCTURE OF THE 50
H57 AND GAS 5 MRNAS
3.2.4.2 EXPRESSION OF H57 MRNA IS CONSISTENT 50
WITH A GROWTH ARREST ASSOCIATED GENE
3.3 EXPRESSION CLONING OF THE CDNA ENCODING 51
THE EPITOPE RECOGNISED BY THE ELL ANTIGEN
3.3.1 CLONING STRATEGY 51
3.32 IMMUNOPRECIPITATION OF PUTATIVE ELL PROTEIN 52
33.3 TISSUE DISTRIBUTION OF THE ELL ANTIGEN EXPRESSION 52
3.3.4 ANALYSIS OF ELL PROTEIN SEQUENCE ' 53
3.4 APOPTOSIS AS THE TERMINAL DIFFERENTIATION STEP IN 54
THE OSTEOBLASTIC LINEAGE AND AS A POTENTIAL TARGET FOR
THE THERAPY OF OSTEOPOROSIS
3.4.1 APOPTOTIC CELL DEATH OF ROS 17/2.8 CELLS 54
3.42 SELECTIVE LOSS OF THE OSTEOBLASTIC PHENOTYPE PRECEDES
56
APOPTOSIS OF ROS 17/2.8 CELLS IN VITRO
3.4.3 INDUCTION OF APOPTOSIS BY MISTLETOE LECTIN 58
3.4.4 THERAPEUTIC APPROACHES TO MODULATION OF APOPTOSIS
59
4 DISCUSSION 62
4.1 IDENTIFICATION OF MARKERS OF OSTEOBLAST DIFFERENTIATION 62
4.1.1 THYMOSIN P 4 63
4.12 FERRITIN
64
4.1.3. H57 (GAS-5)
65
4.1.4 NOVEL ESTS
66
4.15 ELL ANTIGEN
67
4.2 APOPTOSIS AND THE THERAPY OF OSTEOPOROSIS 68
4.2.1 OSTEOBLASTIC CELLS DIE BY APOPTOSIS 68
4.2.2 PHENOTYPIC LOSS AND APOPTOSIS 69
4.2.3 OSTEOBLAST APOPTOSIS IS INDUCED BY CYTOTOXIC MISTLETOE 69
LECTIN
4.2.4 SURVIVAL FACTORS AND APOPTOSIS OF OSTEOBLASTS 70
5 CONCLUSIONS 72
6 SUMMARY 74
7 REFERENCES 75 |
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| physical | III, 87 Bl. Ill., graph. Darst. |
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| spelling | Atkinson, Michael John Verfasser (DE-588)131634100 aut Molecular biological studies into the pathophysiology of osteoporosis vorgelegt von Michael J. Atkinson 1998 III, 87 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Habil.-Schr., 1998 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008715505&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Atkinson, Michael John Molecular biological studies into the pathophysiology of osteoporosis |
| subject_GND | (DE-588)4113937-9 |
| title | Molecular biological studies into the pathophysiology of osteoporosis |
| title_auth | Molecular biological studies into the pathophysiology of osteoporosis |
| title_exact_search | Molecular biological studies into the pathophysiology of osteoporosis |
| title_full | Molecular biological studies into the pathophysiology of osteoporosis vorgelegt von Michael J. Atkinson |
| title_fullStr | Molecular biological studies into the pathophysiology of osteoporosis vorgelegt von Michael J. Atkinson |
| title_full_unstemmed | Molecular biological studies into the pathophysiology of osteoporosis vorgelegt von Michael J. Atkinson |
| title_short | Molecular biological studies into the pathophysiology of osteoporosis |
| title_sort | molecular biological studies into the pathophysiology of osteoporosis |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008715505&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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