Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS):
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English German |
| Published: |
2000
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Erlangen-Nürnberg, Univ., Diss., 2000. - Zsfassung in dt. Sprache |
| Physical Description: | VIII, 78 S. Ill., graph. Darst. |
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| 100 | 1 | |a Qadoumi, Muna |d 1970- |e Verfasser |0 (DE-588)12273744X |4 aut | |
| 245 | 1 | 0 | |a Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) |c by Muna Qadoumi |
| 264 | 1 | |c 2000 | |
| 300 | |a VIII, 78 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Erlangen-Nürnberg, Univ., Diss., 2000. - Zsfassung in dt. Sprache | ||
| 502 | |a Erlangen-Nürnberg, Univ., Diss., 2000 | ||
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009302052 | |
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| _version_ | 1807504479034015744 |
|---|---|
| adam_text |
LIST
OF
CONTENTS
PAGE
LIST
OF
ABBREVIATIONS
I
LIST
OF
FIGURES
IV
LIST
OF
TABLES
V
ABSTRACT
VI
I.
INTRODUCTION
1
1.
NOS
FAMILY
AND
NO
PRODUCTION
1
2.
INOS
EXPRESSION
1
3.
REGULATION
OF
INOS
EXPRESSION
:
INDUCTION
AND
INHIBITION
OF
MURINE
INOS
2
4.
NO
FUNCTION
IN
THE
IMMUNE
SYSTEM
5
5.
THE
ANTIMICROBIAL
FUNCTION
OF
NO
IN
MOUSE
INFECTION
MODELS
5
6.
THE
FUNCTION
OF
NO
IN
THE
LEISHMANIA
MOUSE
MODEL
6
7.
EVIDENCE
FOR
NO-RELATED
ANTIMICROBIAL
ACTIVITIES
IN
HUMANS
8
II.
OBJECTIVES
OF
THE
STUDY
IO
III.
MATERIALS
AND
METHODS
1
1
A)
IN
VITRO
STUDIES
11
1.
CELLS
AND
CELL
LINES
11
2.
CELL
CULTURE
12
2.1.
MEDIA
12
2.2.
CULTURE
12
2.3.
CRYOPRESERVATION
13
PAGE
3.
CELL
VIABILITY
ASSAY
13
4.
CYTOKINES
AND
CELL
STIMULATION
14
5.
DETERMINATION
OF
NITRITE
(NO
2
)
ACCUMULATION
IN
CULTURE
SUPERNATANTS
(NO-ASSAY)
14
6.
PREPARATION
OF
CELL
LYSATES
FOR
WESTERN
BLOT
ANALYSIS
15
6.
1
.
PREPARATION
OF
LYSATES
15
6.2.
DETERMINATION
OF
PROTEIN
CONCENTRATION-BRADFORD
METHOD
15
7.
PREPARATION
OF
CELL
LYSATES
FOR
IMMUNOPRECIPITATION
15
7.1.
PREPARATION
OF
LYSATES
15
7.2.
DETERMINATION
OF
PROTEIN
CONCENTRATION-PIERCE
METHOD
15
8.
PREPARATION
OF
NUCLEAR
EXTRACTS
16
9.
IMMUNOPRECIPITATION
OF
SIGNAL
TRANSDUCER
AND
ACTIVATOR
OF
TRANSCRIPTION
16
6
(STAT6)
PROTEIN
10.
SODIUM
DODECYL
SULFATE-POLYACRYLAMIDE
GEL-ELECTROPHORESIS
(SDS-PAGE)
17
11.
IMMUNOBLOT
ANALYSIS
17
11
.
1
.
DETECTION
OF
HUMAN
INOS
PROTEIN
1
7
1
1
.2.
DETECTION
OF
HUMAN
STAT6
PROTEIN
18
12.
RNA-PREPARATION
18
13.
REVERSE
TRANSCRIPTION
19
14.
POLYMERASE
CHAIN
REACTION
(PCR)
19
14.
1
.
QUALITATIVE
PCR
19
14.2.
COMPETITIVE
QUANTITATIVE
REVERSE
TRANSCRIPTASE
PCR
20
14.3.
GENERATION
OF
DNA-COMPETITORS
20
14.4.
OLIGONUCLEOTIDE
PRIMERS
21
15.
SUBCLONING
OF
PCR
PRODUCTS
15.1.
ELECTROELUTION
OF
CDNA
PCR
PRODUCTS
FROM
AGAROSE
GEL
22
PAGE
15.2.
LIGATION
OF
DNA
FRAGMENTS
WITH
LINEARIZED
PLASMID
VECTORS
22
15.3.
PREPARATION
OF
COMPETENT
E.
COLL
CELLS
23
15.4.
TRANSFORMATION
OF
BACTERIA
WITH
PLASMID
DNA
23
15.4.1
TRANSFORMATION
BY
ELECTROPORATION
23
15.4.2
TRANSFORMATION
BY
HEAT
SHOCK
24
15.5.
SCREENING
OF
TRANSFORMED
CELLS
FOR
DNA
INSERT
24
15.6.
ISOLATION
OF
PLASMID
DNA
24
16.
INTRODUCTION
OF
DNA
INTO
MAMMALIAN
CELLS:
TRANSIENT
TRANSFECTION
25
16.1.
HINOS
PROMOTER
CONSTRUCTS
AND
LUCIFERASE
REPORTER
SYSTEMS
25
16.2.
TRANSFECTION
METHODS
25
16.2.1
ELECTROPORATION
25
16.2.2
EFFECTENE
1
"'
TRANSFECTION
REAGENT
26
16.2.3
SUPERFECT*
TRANSFECTION
REAGENT
26
16.2.4
DOTAP"'TRANSFECTION
REAGENT
26
16.3.
LUCIFERASE
REPORTER
ASSAY
SYSTEMS
27
17.
APPENDIX
1:
REAGENTS
AND
SOLUTIONS
27
B)
IN
SITU
STUDIES
30
1.
PATIENTS
30
1.1.
PATIENTS
WITH
RHEUMATOID
ARTHRITIS
DISEASE
30
1.2.
PATIENTS
WITH
LOCAL
OR
DIFFUSE
CUTANEOUS
LEISHMANIOSIS
31
2.
STAINING
OF
SINGLE
ANTIGENS
32
3.
IMMUNOFLUORESCENCE
LABELING
OF
TISSUE
SECTIONS
33
4.
EVALUATION
OF
SINGLE
ANTIGEN
STAINING
34
PAGE
IV.
RESULTS
35
A)
IN
VITRO
STUDIES
35
1.
RT4
CELLS
PRODUCE
HIGH
LEVELS
OF
NO
AFTER
STIMULATION
WITH
35
DIFFERENT
CYTOKINE
COMBINATIONS
BUT
NOT
WITH
IFN-Y/LPS
2.
STIMULATION
WITH
IFN-Y/IL-IP
REDUCES
MITOCHONDRIAL
ACTIVITY
37
OF
RT4
CELLS
:
EVIDENCE
FOR
CYTOTOXICITY
3.
NEITHER
TGF-P
NOR
IL
13
OR
IL-4
INHIBITES
CYTOKINE-MEDIATED
NO
37
RELEASE
IN
RT4
CELLS
4.
IL
13
IN
COMBINATION
WITH
IL
1
P
INDUCES
RT4
CELLS
FOR
NO
PRODUCTION
39
5.
IL
1
3/IL-IP-MEDIATED
NO
RELEASE
BY
RT4
CELLS
IS
DOSE
DEPENDENT
39
6.
IL
1
3/IL
1
P
INDUCE
INOS
PROTEIN
AND
MRNA
EXPRESSION
IN
RT4
CELLS
40
7.
KINETICS
OF
CYTOKINE-MEDIATED
NO
PRODUCTION
AND
INOS
EXPRESSION
42
IN
RT4
CELLS
8.
IL-13/IL-IP
INDUCE
INOS
EXPRESSION
AND
NO
PRODUCTION
IN
RT4
CELLS,
42
BUT
NOT
IN
OTHER
HUMAN
CELLS
9.
RT4
CELLS
EXPRESS
TYPE
I
AND
TYPE
II
IL
13
RECEPTOR
45
10.
IL
13
ACTIVATES
STAT6
PROTEIN
IN
RT4
47
B)
IN
SITU
STUDIES
48
I.
INOS
MRN
A
EXPRESSION
IN
SYNOVIAL
TISSUE
FROM
PATIENTS
WITH
RA
48
2.
INOS
PROTEIN
EXPRESSION
IN
SKIN
LESIONS
OF
PATIENTS
WITH
LCL
OR
DCL
49
V.
DISCUSSION
52
1.
REGULATION
OF
INOS
IN
RT4
CELLS
52
1.1.
CYTOKINE-MEDIATED
INOS
INDUCTION
IN
RT4
CELLS
52
PAGE
1.2.
HYPORESPONSIVENESS
OF
THE
HUMAN
INOS
GENE
TO
STIMULATION
BY
54
IFN-Y/LPS
1.3.
IFN-A/IFN-P-MEDIATED
NO
PRODUCTION
IN
RT4
CELLS
54
1.4.
THE
EFFECT
OF
TH2-DERIVED
CYTOKINES
ON
NO
PRODUCTION
IN
RT4
CELLS
55
1.5.
RT4
CELLS
AND
IL
13
55
1.5.1.
SYNTHESIS
AND
FUNCTION
OF
IL-1
3
55
1.5.2.
IL-13
MEDIATES
INOS
EXPRESSION
AND
NO
PRODUCTION
IN
RT4
CELLS,
56
BUT
NOT
IN
OTHER
HUMAN
CELLS
1.5.3.
IL
13
RECEPTOR
SYSTEMS
57
1.5.4.
RT4
CELLS
EXPRESS
TYPE
I
AND
TYPE
II
IL
1
3R
57
1.5.5.
IL-13-MEDIATED
STAT6
ACTIVATION
59
2.
EXPRESSION
OF
HUMAN
INOS
IN
VIVO
60
2.1.
DETECTION
OF
INOS
MRNA
IN
SYNOVIAL
TISSUE
OF
PATIENTS
WITH
RA
60
2.2.
DETECTION
OF
INOS
PROTEIN
IN
SKIN
LESIONS
OF
PATIENTS
WITH
ACL
60
VI.
CONCLUSION
63
VII.
REFERENCES
64 |
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| author_GND | (DE-588)12273744X |
| author_facet | Qadoumi, Muna 1970- |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV013615146 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T01:09:56Z |
| institution | BVB |
| language | English German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009302052 |
| oclc_num | 48742767 |
| open_access_boolean | |
| owner | DE-29 DE-29T DE-355 DE-BY-UBR DE-12 DE-83 DE-188 |
| owner_facet | DE-29 DE-29T DE-355 DE-BY-UBR DE-12 DE-83 DE-188 |
| physical | VIII, 78 S. Ill., graph. Darst. |
| publishDate | 2000 |
| publishDateSearch | 2000 |
| publishDateSort | 2000 |
| record_format | marc |
| spelling | Qadoumi, Muna 1970- Verfasser (DE-588)12273744X aut Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) by Muna Qadoumi 2000 VIII, 78 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2000. - Zsfassung in dt. Sprache Erlangen-Nürnberg, Univ., Diss., 2000 Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd rswk-swf Stickstoffmonoxid-Synthase (DE-588)4481118-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Stickstoffmonoxid-Synthase (DE-588)4481118-4 s Genregulation (DE-588)4122166-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009302052&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Qadoumi, Muna 1970- Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd Stickstoffmonoxid-Synthase (DE-588)4481118-4 gnd |
| subject_GND | (DE-588)4122166-7 (DE-588)4481118-4 (DE-588)4113937-9 |
| title | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) |
| title_auth | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) |
| title_exact_search | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) |
| title_full | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) by Muna Qadoumi |
| title_fullStr | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) by Muna Qadoumi |
| title_full_unstemmed | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) by Muna Qadoumi |
| title_short | Tissue expression and cytokine regulation of the human inducible nitric oxide synthase (iNOS) |
| title_sort | tissue expression and cytokine regulation of the human inducible nitric oxide synthase inos |
| topic | Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd Stickstoffmonoxid-Synthase (DE-588)4481118-4 gnd |
| topic_facet | Hochschulschrift Genregulation Stickstoffmonoxid-Synthase |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009302052&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT qadoumimuna tissueexpressionandcytokineregulationofthehumaninduciblenitricoxidesynthaseinos |