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Showing 1 to 4 of 4 (0.035 seconds)Spelling suggestions: "subject:"MALDI- TOF- TOF/massspectrometry"" "subject:"MALDI- TOF- TOF/masspectrometry""
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Identification of human hair follicle antigens targeted in the presumptive autoimmune hair follicle disorder Alopecia Areata and their potential functional relevance In Vitro. Methods development for isolation and identification of Alopecia Areata-relevant human hair follicle antigens using a proteomics approach and their functional assessment using an Ex Vivo hair follicle organ culture model.Leung, Man Ching January 2008 (has links)
Alopecia areata (AA) is a putative autoimmune hair loss disorder. It mainly affects the scalp hair but can also involve body hair, and can also affect the nail and the eye. While there are may be several lines of evidence to support the autoimmune basis of AA, there is still very little information on the hair follicle autoantigen(s) involved in its pathogenesis.
In this project, serum antibodies (AA=10, control=10) were used to immunoprecipitate AA-relevant target antigens from normal human scalp hair follicle extracts. These immunoprecipitates were analysed by LC-MALDI-TOF/TOF mass spectrometry for target protein identification. This part of the project involved substantial methods development.
Trichohyalin was immunoprecipitated by all AA sera, but by only 5 normal sera. Importantly, the mean Mascot scores of the AA group was significantly higher than the normal group (p=0.005). Keratin 16 was also identified from immunoprecipitates as another potential AA-relevant target antigen.
Functional studies by ex vivo whole hair follicle organ culture using commercial antibodies to trichohyalin and keratin 16 significantly inhibited hair fibre elongation compared to controls.
Indirect immunofluorescence studies revealed that AA sera contained higher immunoreactivity against normal human scalp anagen hair follicles compared to normal sera. Immunoreactivities were mainly in the outer root sheath and inner root sheath, and less so to the medulla and hair bulb matrix. Double immunofluorescence studies of AA and normal serum with anti-trichohyalin antibody (AE15) revealed co-localisation of 9 of the AA sera antibodies with trichohyalin in the inner root sheath (mostly in Henle¿s, less in Huxley¿s/inner root sheath cuticle), but only weakly in 3 normal sera.
This study supports the involvement of an antibody response to anagen-specific hair follicles antigens in AA. Moreover, there may be some evidence that these antibodies may have a pathogenic role.
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Identification of human hair follicle antigens targeted in the presumptive autoimmune hair follicle disorder alopecia areata and their potential functional relevance in vitro : methods development for isolation and identification of alopecia areata-relevant human hair follicle antigens using a proteomics approach and their functional assessment using an ex vivo hair follicle organ culture modelLeung, Man Ching January 2008 (has links)
Alopecia areata (AA) is a putative autoimmune hair loss disorder. It mainly affects the scalp hair but can also involve body hair, and can also affect the nail and the eye. While there are may be several lines of evidence to support the autoimmune basis of AA, there is still very little information on the hair follicle autoantigen(s) involved in its pathogenesis. In this project, serum antibodies (AA=10, control=10) were used to immunoprecipitate AA-relevant target antigens from normal human scalp hair follicle extracts. These immunoprecipitates were analysed by LC-MALDI-TOF/TOF mass spectrometry for target protein identification. This part of the project involved substantial methods development. Trichohyalin was immunoprecipitated by all AA sera, but by only 5 normal sera. Importantly, the mean Mascot scores of the AA group was significantly higher than the normal group (p=0.005). Keratin 16 was also identified from immunoprecipitates as another potential AA-relevant target antigen. Functional studies by ex vivo whole hair follicle organ culture using commercial antibodies to trichohyalin and keratin 16 significantly inhibited hair fibre elongation compared to controls. Indirect immunofluorescence studies revealed that AA sera contained higher immunoreactivity against normal human scalp anagen hair follicles compared to normal sera. Immunoreactivities were mainly in the outer root sheath and inner root sheath, and less so to the medulla and hair bulb matrix. Double immunofluorescence studies of AA and normal serum with anti-trichohyalin antibody (AE15) revealed co-localisation of 9 of the AA sera antibodies with trichohyalin in the inner root sheath (mostly in Henle's, less in Huxley's/inner root sheath cuticle), but only weakly in 3 normal sera. This study supports the involvement of an antibody response to anagen-specific hair follicles antigens in AA. Moreover, there may be some evidence that these antibodies may have a pathogenic role.
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Genomic and proteomic analysis of drought tolerance in Sorghum (Sorghum bicolor (L.) Moench)Woldesemayat, Adunga,Abdi January 2014 (has links)
Philosophiae Doctor - PhD / Drought is the most complex phenomenon that remained to be a potential and historic challenge to
human welfare. It affects plant productivity by eliciting perturbations related to a pathway that
controls a normal, functionally intact biological process of the plant. Sorghum (Sorghum bicolor
(L.) Moench), a drought adapted model cereal grass is a potential target in the modem agricultural
research towards understanding the molecular and cellular basis of drought tolerance. This study
reports on the genomic and proteomic findings of drought tolerance in sorghum combining the
results from in silica and experimental analysis. Pipeline that includes mapping expression data
from 92 normalized cDNAs to genomic loci were used to identify drought tolerant genes. Integrative analysis was carried out using sequence similarity search, metabolic pathway, gene expression profiling and orthology relation to investigate genes of interest. Gene structure prediction was conducted using combination of ab initio and extrinsic evidence-driven information employing multi-criteria sources to improve accuracy. Gene ontology was used to cross-validate and to functionally assign and enrich genes. An integrated approach that subtly combines functional ontology based semantic data with
expression profiling and biological networks was employed to analyse gene association with plant
phenotypes and to identify and genetically dissect complex drought tolerance in sorghum. The
gramene database was used to identify genes with direct or indirect association to drought related
ontology terms in sorghum. Where direct association for sorghum genes were not available, genes
were captured using Ensemble Biomart by transitive association based on the putative functions of
sorghum orthologs in closely related species. Ontology mapping represented a direct or transitive
association of genes to multiple drought related ontology terms based on sorghum specific genes or
orthologs in related species. Correlation of genes to enriched gene ontology (GO)-terms (p-value <
0.05) related to the whole-plant structure was used to determine the extent of gene-phynotype association across-species and environmental stresses.
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Die Entwicklung von Immunoproteomics-Methoden am Beispiel der Identifizierung Magenkarzinom-assoziierter Helicobacter pylori AntigeneKrah, Alexander 20 December 2004 (has links)
Das magenbesiedelnde Bakterium Helicobacter pylori gehört zu den am weitesten verbreiteten Infektionserregern. Obwohl die Infektion meist lebenslang symptomlos verläuft, kann H. pylori bei einigen Menschen schwere Erkrankungen bis hin zum Magenkarzinom verursachen. Ziele dieser Arbeit waren Magenkarzinom-assoziierte Antigene für einen diagnostischen Test zu finden und Methoden zur Untersuchung von Spotkompositionen mittels MALDI-TOF/TOF Massenspektrometrie zu entwickeln. Im ersten Teil der Promotion wurden die Antigenerkennungsmuster von 30 Magenadenokarzinom- mit 30 Ulkus duodeni-Patienten mithilfe hochauflösender zweidimensionaler Immunoblots von H. pylori Lysat verglichen. Diese fokussierte Gegenüberstellung eignet sich gut für diese Fragestellung, da beide Erkrankungen von diesem Bakterium verursacht werden, aber nur sehr selten gemeinsam auftreten. Durch univariate statistische Analysen wurden 14 Magenkarzinom korrelierte Spots gefunden (p / The stomach-colonizing bacterium Helicobacter pylori is one of the most widespread infectious agents. Although infection mostly persists unnoticed, it may cause serious diseases like gastric carcinoma. Aims of this project were to find gastric carcinoma-associated antigens for a diagnostic test and to develop methods to analyze spot compositions using MALDI-TOF/TOF mass spectrometry. In the first part of this project antigen recognition patterns of 30 gastric carcinoma- and 30 duodenal ulcer- patients were compared using high-resolution two-dimensional immunoblots of H. pylori lysate. This focused comparison lent itself to this question because both diseases are caused by the bacterium but rarely occur conjointly. Utilizing univariate statistical tests 14 gastric carcinoma-associated spots were found (p
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