Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Leipzig, Univ., Diss., 2002 |
| Beschreibung: | VI, 125, [13] Bl. Ill., graph. Darst. : 30 cm |
Internformat
MARC
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| 100 | 1 | |a Petković, Marijana |d 1971- |e Verfasser |0 (DE-588)124350585 |4 aut | |
| 245 | 1 | 0 | |a Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |c vorgelegt von Marijana Petković |
| 246 | 1 | |a Role of lipid-derived second messengers in the oxidative activity of human polymorphonuclear leukocytes | |
| 264 | 1 | |c 2002 | |
| 300 | |a VI, 125, [13] Bl. |b Ill., graph. Darst. : 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
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Datensatz im Suchindex
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| adam_text |
TABLE OF CONTENTS
1. INTRODUCTION 1
1.1. POLYMORPHONUCLEAR LEUKOCYTES 1
1.1.1. NEUTROPHI 1 BACTERICIDAL AGENTS 3
1.1.2. OXIDATIVE ACTIVITY OF NEUTROPHILS 5
1.1.2.1. REACTIVE OXYGEN METABOLITES PRODUCED BY STIMULATED HUMAN
NEUTROPHILS 6
1.1.2.2. METHODS FOR THE DETECTION OF REACTIVE OXYGEN SPECIES 7
1.2. SIGNALLING PATHWAYS IN HUMAN NEUTROPHILS 8
1.2.1. SIGNALLING PATHWAYS INITIATED BY CHEMOTACTIC TRIPEPTIDE-FMLP 8
1.2.1.1. ROLE OF LIPID-DERIVED SECOND MESSENGERS IN THE ACTIVATION OF
NEUTROPHILS 10
1.2.1.1.1. PHOSPHATIDIC ACIDS 10
1.2.1.1.2. LYSOPHOSPHOLIPIDS . 11
1.2.1.1.3. PHOSPHOLIPASE A
2
12
1.3. LIPID AND PHOSPHOLIPID ANALYSIS 14
1.3.1. MASS SPECTROMETRIC METHODS IN LIPID ANALYSIS 15
1.3.2. APPLICATION OF MALDI-TOF MS FOR THE LIPID ANALYSIS 15
1.3.3. DETERMINATION OF PHOSPHOLIPASE A
2
ACTIVITY 16
1.4. AIM OF THE STUDY 18
2. MATERIALS AND METHODS 19
2.1. MATERIALS 19
2.1.1. LIPIDS AND PHOSPHOLIPIDS 19
2.1.2. CHEMICALS FOR THE PURIFICATION OF NEUTROPHILS AND FOR
CHEMILUMINESCENCE
MEASUREMENTS 19
2.1.3 CHEMICALS FOR MALDI-TOF MASS SPECTROMETRY 22
2.1.4. CHEMICALS FOR POLYACRYLAMIDE GEL ELECTROPHORESIS 22
2.2.
EXPERIMENTS WITH POLYMORPHONUCLEAR LEUKOCYTES 22
2.2.1. ISOLATION OF HUMAN POLYMORPHONUCLEAR LEUKOCYTES 22
2.2.2. CHEMILUMINESCENCE MEASUREMENTS 23
2.2.3. EXTRACTION OF LIPIDS FROM HUMAN NEUTROPHILS / 25
2.2.4. INCUBATION OF NEUTROPHILS WITH PHOSPHOLIPID VESICLES 26
2.2.5. FLUORESCENCE MICROSCOPY OF HUMAN NEUTROPHILS 26
2.3. MALDI-TOF MS OF PHOSPHOLIPIDS 27
2.3.1. SAMPLE AND MATRIX PREPARATION 27
2.3.2. DIGESTION OF NATURAL PHOSPHOLIPIDS WITH PANCREATIC PHOSPHOLIPASE
A
2
27
2.3.3. EQUIPMENT FOR MALDI-TOF MASS SPECTROMETRY 28
2.3.4. QUANTIFICATION OF SELECTED LIPIDS BY MALDI-TOF MS 28
2.4. EXPERIMENTS WITH PHOSPHOLIPASE
HI
29
2.4.1. EXTRACTION OF PHOSPHOLIPASE
HI
FROM HUMAN NEUTROPHILS 29
2.4.2. DETERMINATION OF THE PROTEIN CONCENTRATION 29
2.4.3. GEL ELECTROPHORESIS 30
I
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/966167783
2.4.3.1. STAINING OF PROTEINS IN A POLYACRYLAMIDE GEL: COOMASSIE
BRILLIANT BLUE 30
2.4.3.2. STAINING OF PROTEINS IN A POLYACRYLAMIDE GEL: SILVER STAINING
30
2.4.4. MALDI-TOF MASS SPECTROMETRY OF PROTEINS 31
2.4.5 DIGESTION OF PHOSPHATIDYLCHOLINE WITH PANCREATIC PHOSPHOLIPASE A
2
31
2.4.6. INCUBATION OF PHOSPHOLIPIDS WITH PHOSPHOLIPASE A
2
EXTRACTED FROM
HUMAN NEUTROPHILS AND WITH THE SUPERNATANT OF NEUTROPHILS 32
2.4.7. DETERMINATION OF THE PHOSPHOLIPASE A2 ACTIVITY BY COLORIMETRIC
ASSAY 32
3. RESULTS AND DISCUSSION 34
3.1. MALDI-TOF MASS SPECTROMETRY OF PHOSPHOLIPIDS 34
3.1.1. CHARACTERISATION OF THE MALDI-TOF MASS SPECTRA OF MATRIX (2,5-
DIHYDROXYBENZOIC ACID) 35
3.1.2. GENERAL PROPERTIES OF MALDI-TOF MASS SPECTRA OF PHOSPHOLIPIDS 36
3.1.3. POSITIVE ION MALDI-TOF MASS SPECTRA OF DIFFERENT PHOSPHOLIPID 38
SPECIES
3.1.4. NEGATIVE ION MALDI-TOF MASS SPECTRA OF PHOSPHOLIPIDS 41
3.1.5. MALDI-TOF MASS SPECTRA OF LYSOPHOSPHOLIPIDS 43
3.1.5.1. MALDI-TOF MASS SPECTRA OF NATURAL LYSOPHOSPHOLIPID MIXTURES 45
3.1.6. MALDI-TOF MASS SPECTRA OF COMPLEX PHOSPHOLIPID AND
LYSOPHOSPHOLIPID MIXTURES 47
3.1.7. QUANTIFICATION OF LYSOPHOSPHOLIPIDS BY MALDI-TOF MASS
SPECTROMETRY 51
3.1.7.1. DETERMINATION OF THE LPC (18:0) CONTENT IN LIPID EXTRACS OF
HUMAN
POLYMORPHONUCLEAR LEUKOCYTES 55
3.1.8. SUMMARY OF MALDI-TOF MASS SPECTROMETRY OF PHOSPHOLIPIDS 57
3.2 OXIDATIVE ACTIVITY OF HUMAN POLYMORPHONUCLEAR LEUKOCYTES 58
3.2.1. ROLE OF SELECTED ENZYMES IN THE ACTIVATION OF NEUTROPHIL NADPH 58
OXIDASE 1
3.2.1.1. INHIBITION OF PHOSPHATIDYLINOSITOL 3-KINASE 59
3.2.1.2. INHIBITION OF CYTOSOLIC PHOSPHOLIPASE A
2
60
3.2.1.3. INHIBITION OF DIACYLGLYCEROL KINASE 61
3.2.2. INVESTIGATION OF THE LIPID COMPOSITION OF HUMAN NEUTROPHILS BY
MALDI-TOF MASS SPECTROMETRY / 62
3.2.2.1. INVESTIGATION OF THE LYSOPHOSPHOLIPIDS IN THE ORGANIC EXTRACTS
OF
HUMAN NEUTROPHILS UNDER DIFFERENT STIMULATION CONDITIONS 64
3.2.3. EFFECTS OF EXOGENOUSLY-ADDED LYSOPHOSPHOLIPIDS ON THE OXIDATIVE
ACTIVITY OF HUMAN NEUTROPHILS 67
3.2.3.1. INFLUENCE OF THE EXOGENOUS LYSOPHOSPHOLIPIDS ON THE
CHEMILUMINESCENCE RESPONSE OF HUMAN NEUTROPHILS 68
3.2.3.2. MALDI-TOF MS OF THE LIPID EXTRACTS OF HUMAN NEUTROPHILS 71
3.2.4. EFFECTS OF EXOGENOUSLY-ADDED LONG CHAIN PHOSPHATIDIC ACIDS ON THE
OXIDATIVE ACTIVITY OF HUMAN NEUTROPHILS 74
3.2.4.1. MALDI-TOF MS OF THE ORGANIC EXTRACTS OF NEUTROPHILS AFTER
STIMULATION WITH PHOSPHATIDIC ACIDS 75
II
3.2.4.2. MODULATION OF THE CHEMILUMINESCENCE RESPONSE OF PHOSPHATIDIC
ACID-
STIMULATED HUMAN NEUTROPHILS 76
3.2.4.3. MICROSCOPIC INVESTIGATIONS OF THE INTERACTION OF PHOSPHATIDIC
ACIDS
WITH NEUTROPHILS 79
3.2.5. SUMMARY OF THE EFFECTS OF PHOSPHOLIPIDS ON THE OXIDATIVE ACTIVITY
OF
HUMAN NEUTROPHILS 81
3.3. DETECTION AND ANALYSIS OF THE PHOSPHOLIPASE A
2 ACTIVITY SECRETED FROM
HUMAN NEUTROPHILS ' 83
3.3.1. APPLICATION OF MALDI-TOF MASS SPECTROMETRY FOR MONITORING THE
KINETICS OF PHOSPHATIDYLCHOLINE DIGESTION BY PANCREATIC PHOSPHOLIPASE A2
83
3.3.1.1. INFLUENCE OF THE DETERGENT ON THE PPLA2 ACTIVITY 85
3.3.1.2. KINETICS OF THE PHOSPHATIDYLCHOLINE DIGESTION WITH
PHOSPHOLIPASE A
2 86
3.3.1.3. DETERMINATION OF THE LPC CONCENTRATION FROM THE MALDI-TOF MASS
SPECTRA 88
3.3.1.4. INFLUENCE OF CA
2+ AND PC CONCENTRATION ON THE LPC GENERATION _ 89
3.3.2. MALDI-TOF MASS SPECTROMETRY OF PROTEINS SECRETED FROM HUMAN
NEUTROPHILS AND THE SAMPLE OBTAINED BY THE ACIDIC EXTRACTION PROCEDURE
91
3.3.2.2. ANALYSIS OF THE SAMPLE OBTAINED BY THE ACID EXTRACTION
PROCEDURE FROM
HUMAN NEUTROPHILS 92
3.3.2.3. MALDI-TOF MASS SPECTROMETRIC ANALYSIS OF THE PROTEINS SECRETED
FROM HUMAN NEUTROPHILS 94
3.3.2.4. DETECTION OF THE PHOSPHOLIPASE A
2 ACTIVITY IN THE SUPERNATANT OF THE
HUMAN NEUTROPHILS AND IN THE SAMPLE OBTAINED BY THE ACIDIC EXTRACTION
PROCEDURE 96
3.3.2.4.1. DETECTION OF PHOSPHOLIPASE A
2 ACTIVITY IN THE SAMPLE OBTAINED BY THE
ACIDIC EXTRACTION PROCEDURE FROM THE NEUTROPHILS 97
3.3.2.4.2. DETECTION OF THE PHOSPHOLIPASE A
2 ACTIVITY SECRETED FROM HUMAN
NEUTROPHILS 98
3.3.3. SUMMARY OF THE DETERMINATION OF THE PHOSPHOLIPASE A
2 ACTIVITY SECRETED
FROM HUMAN NEUTROPHILS 100
4. SUMMARY AND CONCLUSIONS 101
5. REFERENCES 107
SELBSTSTANDIGKEITSERKLARUNG
CURRICULUM VITAE
PUBLICATION LIST
III |
| any_adam_object | 1 |
| author | Petković, Marijana 1971- |
| author_GND | (DE-588)124350585 |
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| author_role | aut |
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| bvnumber | BV016553854 |
| ctrlnum | (OCoLC)633425007 (DE-599)BVBBV016553854 |
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| illustrated | Illustrated |
| indexdate | 2024-11-22T17:36:11Z |
| institution | BVB |
| language | English |
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| physical | VI, 125, [13] Bl. Ill., graph. Darst. : 30 cm |
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| spelling | Petković, Marijana 1971- Verfasser (DE-588)124350585 aut Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes vorgelegt von Marijana Petković Role of lipid-derived second messengers in the oxidative activity of human polymorphonuclear leukocytes 2002 VI, 125, [13] Bl. Ill., graph. Darst. : 30 cm txt rdacontent n rdamedia nc rdacarrier Leipzig, Univ., Diss., 2002 Leukozyt (DE-588)4074195-3 gnd rswk-swf Sekundärer Bote (DE-588)4243447-6 gnd rswk-swf Oxidation (DE-588)4137187-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Leukozyt (DE-588)4074195-3 s Oxidation (DE-588)4137187-2 s Sekundärer Bote (DE-588)4243447-6 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010226058&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Petković, Marijana 1971- Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes Leukozyt (DE-588)4074195-3 gnd Sekundärer Bote (DE-588)4243447-6 gnd Oxidation (DE-588)4137187-2 gnd |
| subject_GND | (DE-588)4074195-3 (DE-588)4243447-6 (DE-588)4137187-2 (DE-588)4113937-9 |
| title | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| title_alt | Role of lipid-derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| title_auth | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| title_exact_search | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| title_full | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes vorgelegt von Marijana Petković |
| title_fullStr | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes vorgelegt von Marijana Petković |
| title_full_unstemmed | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes vorgelegt von Marijana Petković |
| title_short | Role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| title_sort | role of lipid derived second messengers in the oxidative activity of human polymorphonuclear leukocytes |
| topic | Leukozyt (DE-588)4074195-3 gnd Sekundärer Bote (DE-588)4243447-6 gnd Oxidation (DE-588)4137187-2 gnd |
| topic_facet | Leukozyt Sekundärer Bote Oxidation Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010226058&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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