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Rye, Morten Beck. "Image segmentation and multivariate analysis in two-dimensional gel electrophoresis." Doctoral thesis, Norwegian University of Science and Technology, Department of Chemistry, 2007. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-1744.
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The topic of this thesis is data-analysis on images from two-dimensional electrophoretic gels. Because of the complexity of these images, there are numerous steps and approaches to such an analysis, and no “golden standard” has yet been established on how to produce the desired output. In this thesis focus is put on two essential fields concerning 2D-gel analysis; registration of images by segregation and protein spot identification, and data-analysis on the output of such a registration by multivariate methods. Image segmentation is mainly concerned with the task of identifying individual protein spots in a gel-image. This has generally been the natural starting point of all methods and procedures developed since the introduction of 2D-gels in the mid-seventies, simply because this best reproduces the results created by a human analyst, who manually identify protein-spot entities. The amount of data produced in a 2D-gel experiment can be quite large, especially in multiple gels where the human analyst is dependent on additional statistical data-analytical tools to produce results. Because of the correlated nature of most gel-data, analysis by multivariate methods is natural choice, and are therefore adopted in this thesis. The goal of this thesis is to introduce the above mentioned procedures at different stages in the analysis pipeline where they are not yet fully exploited, rather than to improve already existing algorithms. In this way new insight and ideas on how to handle data from 2D-gel experiments are achieved. The thesis starts with a review of segmentation methodology, and introduces a selected procedure used to identify protein spots throughout. Output from the segmentation is then used to create a multivariate spot-filtering model, which aims to separate protein spots from noise and artefacts often creating problems in 2D-gel analysis. Lately the use of common spot boundaries in multiple gels have been the method of choice when gels are analysed. How such boundaries should be defined is an important subject of discussion, and thus a new method for defining common boundaries based on the individual segmentation of each gel is introduced. Segmentation may be a natural starting point when gels are analysed, but it is not necessarily the most correct. Often the introduction of fixed spot entities introduces restrictions to the data which cause problems at later stages in the analysis. Analysing pixels from multiple gels directly has no such restrictions, and it is shown in this thesis that the output of such an analysis based on multivariate methods can produce very useful results. It can also give insight to the data problematic to achieve with the spot boundary approach. At last in the thesis an improved pixel-based approach is introduced, where a less restricted segmentation is used to reduce and concentrate the amount of data analysed, improving the final output.
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Brown, Andrew S. "Two-dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) Characterization of Decorin." Youngstown State University / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1311873768.
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Xu, Aoshuang. "Development in electrophoresis instrumentation for two-dimensional gel electrophoresis of protein separation and application of capillary electrophoresis in micro-bioanalysis /." [Ames, Iowa : Iowa State University], 2008.
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Yao, Mingyi. "Proteomic studies of Pseudomonas putida KT2440 in carcinogenicity screening via 2-dimensional gel electrophoresis /." Online version of thesis, 2005. https://ritdml.rit.edu/dspace/handle/1850/1118.
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Fowlkes, Kelly. "A proteomic study of Pseudomonas putida by two-dimensional gel electrophoresis : establishing quantitative standards for intra-laboratory results /." Online version of thesis, 2007. http://hdl.handle.net/1850/5040.
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Pandey, Archana. "Proteome analysis of Pseudomonas putida KT2440 using 2D gel electrophoresis and LC/ESI-Q-TOF mass spectrometry /." Online version of thesis, 2007. https://ritdml.rit.edu/dspace/handle/1850/3848.
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Rajagopal, Meena Uma. "METHODS DEVELOPMENT AND APPLICATION OF TWO-DIMENSIONAL GEL ELECTROPHORESIS AND MASS SPECTROMETRY IN PROTEOMICS." UKnowledge, 2006. http://uknowledge.uky.edu/gradschool_diss/292.
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The development of a highly sensitive ruthenium-based fluorescent staining solution isdescribed in this dissertation. The in-house synthesized ruthenium complex (RuMS)containing both sulfonated and non-sulfonated ligand has detection limit of 1 ng ofprotein that is better than colloidal coomassie, silver and ruthenium complex containingall sulfonated ligands (RuBPS). RuMS stain has 100-fold dynamic range and does notinterfere with subsequent mass spectral identification of proteins. The capability of inhousesynthesis of the staining solution makes it a viable cost-effective alternative to theexpensive commercially available fluorescent stain, Sypro Ruby. The low detection limit,broad linear dynamic range and compatibility with mass spectrometry, make thedevelopment of this stain a worthwhile pursuit. The staining solution was utilized insubsequent applications of two-dimensional gel electrophoresis (2-DE) technology.Proteomics methodology utilizing 2-DE and mass spectrometry was applied toinvestigate the effect of malathion on the proteome of human neuroblastoma cells.Results indicated that out of 122 proteins that were identified from the neuroblastomaproteome, sixteen proteins were down-regulated while five proteins were significantlyup-regulated after treatment with malathion. Significant down-regulation of calciummodulators like calmodulin and calgizarrin and other key chaperones makes themalathion-treated cells highly prone to oxidative stress. With increased awareness inpesticide related adverse effects, identification of altered proteins in malathion-treatedhuman neuroblastoma cells is a critical finding.Proteomics is a major area of research in the identification of biomarkers for diseases. Anovel immunoprecipitation method developed in this work allowed for successfulisolation and identification of albumin-interactome in cerebrospinal fluid (CSF) that isusually under-represented in standard CSF analysis using 2-DE. A key finding is thedifferential expression of various isoforms of proteins in CSF albumin-interactome fromAlzheimer's disease (AD) subjects. The data implicate the acidic isoform ofprostaglandin D2 synthase (PGDS2) as a potential biomarker for AD. An understandingof the differential expression of these protein isoforms in AD will provide insight into theetiology of the disease and this can have far-reaching implication on drug developmentleading to the cure or even preventation of the disease.
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Ethell, Douglas Wayne. "Analysis of developing chick Gallus domesticus spinal cord proteins using two dimensional gel electrophoresis." Thesis, University of British Columbia, 1990. http://hdl.handle.net/2429/29834.
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Several recent experiments on developing chick spinal cord have established a time window when the developing spinal cord changes from a permissive to a restrictive environment for regeneration. This time window occurs during embryonic days 13-14 (E13-E14) of chick development. Recent experiments in adult rat, have found two proteins that actively inhibit axonal regeneration. This study has sought possible inhibitory proteins, in chicks, correlating to this temporal change. Proteins continuously present after this change (E14-E20) but not before (E11) were identified. Two-dimensional gel electrophoresis was used for separatation of the proteins. Seven protein spots of interest demonstrated this correlative late-expressing neural protein (LNP) profile. Although the functions of these proteins could not be ascertained in this study, further investigation is warranted.Science, Faculty of
Zoology, Department of
Graduate
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Schwartz, Anne. "Characterization of normal and androgen resistant-genital skin fibroblasts using high-resolution two-dimensional gel electrophoresis." Thesis, McGill University, 1985. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=63378.
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Anduri, Sridevi. "Differential protein expression profiles in normal and intersex male smallmouth bass determined using one- and two-dimensional gel electrophoresis a thesis presented to the faculty of the Graduate School, Tennessee Technological University /." Click to access online, 2009. http://proquest.umi.com/pqdweb?index=0&did=2000377671&SrchMode=1&sid=2&Fmt=6&VInst=PROD&VType=PQD&RQT=309&VName=PQD&TS=1277817194&clientId=28564.
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LI, Jinxiang, Ayaka OGASAWARA, Tamao ODAKE, Tomonari UMEMURA, and Kin-ichi TSUNODA. "A New Isoelectric Focusing System for Fast Two-Dimensional Gel Electrophoresis Using a Low-Concentration Polyacrylamide Gel Supported by a Loose Multifilament String." 日本分析化学会, 2005. http://hdl.handle.net/2237/8750.
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Carlsson, Anders. "Identification of potential plasma biomarkers of inflammation in farmers with musculoskeletal disorders : A proteomic study." Thesis, Linköpings universitet, Kemi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110782.
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In this thesis we look for potential chronic inflammation biomarkers because studies have shown that farmers with musculoskeletal disorders might be affected by the environment to develop musculoskeletal disorders. Animal farmers are highly exposed to dust, aerosols, molds and other toxins in the air and environment leading to musculoskeletal disorders, respiratory disorders, airway symptoms and febrile reactions. There is reason to believe that the farmers have a constant or chronic inflammation that develops into musculoskeletal disorders. By using a proteomic approach with Two-dimensional Gel Electrophoresis and silver staining our goal was to find biomarkers by quantifying protein spots that differ significantly from farmers with musculoskeletal disorders compared to rural controls. In our study we found 8 significant proteins, two from Alpha-2-HS-glycoprotein, one from Apolipoprotein A1, three from Haptoglobin, one from Hemopexin and 1 from Antithrombin. All 5 proteins are involved in inflammation response in some way and some proteins are linked to chronic inflammation. Out of the 5 proteins Alpha-2-HS-glycoprotein, Apolipoprotein A1 and Hemopexin seem like the most likely proteins to investigate further as potential inflammation biomarkers.
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Hilson, Tiffany D. "Developing and Optimizing a Mass Spec-Friendly Protocol for Fractionation of tRNAs Through Two-Dimensional Polyacrylamide Gel Electrophoresis." University of Cincinnati / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1322050702.
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Charoensri, Nicha. "Investigation into genetically programmed responses to cadmium and mercury in HeLa cells by differential display and two-dimensional gel electrophoresis." Thesis, McGill University, 2004. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=84491.
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Cadmium and mercury are among the most toxic metals and can be a serious threat to human health. A clear understanding how cells respond, especially in terms of genetically controlled responses to these two metals, is required in order to develop appropriate prevention and treatment procedures that can protect humans from toxic exposure to these metals. Therefore, this study is aimed at elucidating the genetically programmed response to cadmium and mercury exposure in HeLa cells. Two different approaches were employed for this study. Differential expression patterns of mRNAs were studied by differential display reverse transcriptase-polymerase chain reaction (DDRT-PCR) and changes in protein composition of HeLa cells were monitored by two-dimensional (2D) gel electrophoresis. The results showed that transcripts of particular genes were altered by cadmium and/or mercury exposure. These gene are aspartyl/asparginyl beta-hydroxylase (asph), monocyte to macrophage differentiation associated antigen (MMD), and ribosomal protein S24 (rpS24) genes. In addition, some protein spots from 2D gels were found to be altered in their levels as a result of cadmium and/or mercury exposure. The possible roles of asph, MMD and rpS24 genes in response to cadmium and mercury are discussed and further studies are suggested.
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Silva, Fábio Arlindo [UNESP]. "Selênio em tilápia do Nilo utilizando eletroforese em gel e espectrometria atômica." Universidade Estadual Paulista (UNESP), 2009. http://hdl.handle.net/11449/104086.
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silva_fa_dr_botfmvz.pdf: 1538872 bytes, checksum: ed8c4839c4ecda41a8481b80cb1e81f7 (MD5)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Universidade Estadual Paulista (UNESP)
O presente trabalho teve como objetivo investigar a presença de selênio em spots protéicos de amostras de plasma, músculo e fígado de tilápia do Nilo (Oreochromis niloticus) obtidos após separação das proteínas por eletroforese em gel de poliacrilamida em segunda dimensão (2D-PAGE) e posterior avaliação qualitativa por fluorescência de raios-X com radiação síncrotron (SR-XRF). A análise dos espectros de fluorescência obtidos indicaram a presença de selênio em oito proteínas do plasma, seis proteínas do músculo e cinco proteínas do fígado. Observou-se que o selênio está distribuído em sua maioria em proteínas com massa molar menor que 50 kDa. Proteínas acima de 50 kDa foram encontradas somente no plasma.
An investigation was made into selenium in protein spots of samples of plasma, muscle and liver of Nile tilapia (Oreochromis niloticus) obtained after protein separation by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and subsequent qualitative evaluation by synchrotron radiation X-ray fluorescence (SR-XRF). An analysis of the fluorescence spectra indicated the presence of selenium in eight plasma proteins, six muscle proteins, and five liver proteins. Selenium was found to be distributed mainly in proteins with a molar mass smaller than 50 kDa. Proteins with a molar mass higher than 50 kDa was found only in the plasma.
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Bartee, Eric Carter. "Discovery and characterization of a novel family of human ubiquitin ligases termed Membrane Associated RING-CH (MARCH) proteins." Oregon Health & Science University, 2007. http://content.ohsu.edu/u?/etd,629.
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Ph.D.Molecular Microbiology and Immunology
Both poxviruses and γ2-herpesviruses share the K3-family of viral immune evasion proteins. These proteins are characterized by an amino-terminal RING-CH domain followed by two transmembrane domains. We analyzed several human homologues of the K3-family termed membrane-associated RING-CH (MARCH) proteins. All MARCH proteins localized to subcellular membranes while several reduced surface levels of known K3-family substrates. Thus, MARCH proteins appear to be structurally and functionally homologous to viral K3 proteins. One of the major challenges in determining the function of this family is the identification of their physiological substrates. To overcome this we created a quantitative proteomics approach which can be used to identify novel substrates for both the K3- and MARCH-families. Using stable isotope labeling by amino acids in cell culture, we compared the proteome of plasma membrane, golgi, and endoplasmic reticulum membranes in the presence and absence of K5 and MARCH-VIII. Quantitative mass spectrometric protein identification from these fractions revealed that CD316 (bone marrow stromal antigen 2), CD166 (activated leukocyte cell adhesion molecule) and syntaxin-4 were consistently underrepresented in the plasma membrane of K5 expressing cells, while CD44, CD81 (TAPA-1) and B-cell receptor-associated protein 31kDa (Bap31) were consistently underrepresented in the plasma membrane of MARCH-VIII expressing cells. Furthermore, downregulation of each of these proteins was independently confirmed. Our results both identify and characterize a novel family of human ubiquitin ligase enzymes and elucidate a novel technique which can analyze this family and be easily adapted to the analysis of other cellular enzymes viral immune modulators.
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Dennard, Rollin. "Proteomic variations between a Mycoplasma gallisepticum vaccine strain and a virulent field isolate." Digital Archive @ GSU, 2011. http://digitalarchive.gsu.edu/biology_diss/99.
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Mollicutes (mycoplasmas) are pathogenic in a wide range of mammals (including humans), reptiles, fish, arthropods, and plants. Of the medically important mollicutes, Mycoplasma gallisepticum is of particular relevance to avian agriculture and veterinary science, causing chronic respiratory disease in poultry and turkey. Using two-dimensional electrophoresis based quantitative expression proteomics, the current study investigated the molecular mechanisms behind the phenotypic variability between a M. gallisepticum vaccine strain (6/85) and a competitive, virulent field strain (K5234), two strains which were indistinguishable using commonly accepted genetic methods of identification. Twenty-nine proteins showed a significant variation in abundance (fold change > 1.5, p-value < 0.01). Among others, the levels of putative virulence determinants were increased in the virulent K5234, while the levels of several proteins involved with pyruvate metabolism were decreased. It is hoped that the data generated will further the understanding of M. gallisepticum virulence determinants and mechanisms of infection, and that this may contribute to the optimization of diagnostic methodologies and control strategies.
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Badillo-Vargas, Ismael. "Dissecting the molecular interplay between tomato spotted wilt virus and the insect vector, Frankliniella occidentalis." Diss., Kansas State University, 2014. http://hdl.handle.net/2097/35045.
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Doctor of PhilosophyDepartment of Plant Pathology
Anna E. Whitfield
The Bunyaviridae is a family of animal and plant viruses that pose a threat to human, animal, and plant health worldwide. In nature, the dissemination of these viruses is dependent on arthropod vectors (genera Orthobunyavirus, Nairovirus, Phlebovirus, and Tospovirus) or rodent vectors (genus Hantavirus). The genus Tospovirus is the only one within this virus family that is composed of plant-infecting viruses transmitted by thrips. Tomato spotted wilt virus (TSWV), the type species of the Tospovirus genus, is one of the ten most devastating plant viruses known. It is most efficiently transmitted by the western flower thrips, Frankliniella occidentalis Pergande, in a persistant propagative manner. The insect molecules associated with virus infection and transmission by the thrips vector remain unidentified to date. The aim of this work was to identify F. occidentalis larval thrips proteins that are differentially expressed during TSWV infection of the insect vector and those that directly interact with TSWV. To achieve these goals, I used two-dimensional (2-D) gel electrophoresis and mass spectrometry coupled with Mascot searches. I identified 26 protein spots that displayed differential abundances in response to TSWV infection, which contained 37 proteins. Sixty two percent of these proteins were down-regulated by the viral infection demonstrating a complex response. Moreover, 8 and 11 protein spots that directly interacted with purified TSWV virions and a TSWV glycoprotein (GN), respectively, were identified in overlay assays of larval thrips proteins resolved by 2-D gel electrophoresis. A total of five proteins were identified from these spots. These interacting proteins might play roles in attachment and entry, endocytosis/exocytosis, and escape from different tissues for transmission to occur. Injection of double-stranded RNA (dsRNA) into adult female thrips triggered an RNAi response that resulted in 23% reduction of the target gene transcript level. This significant reduction resulted in increased mortality and decreased fertility compared to insects injected with control dsRNA or water and non-injected insects as well. The work presented here provides new insights on the molecular basis of this virus-vector interaction and describes new tools to conduct functional genomic assays to study gene function and design control strategies of F. occidentalis.
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Martin, Kerri Katherine. "Exploring Metallic Flavor Perception: Analysis of Human Salivary Proteins and the Use of the Iron-Binding Protein Lactoferrin in Reducing Metallic Off-Flavors." Thesis, Virginia Tech, 2012. http://hdl.handle.net/10919/76813.
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Metallic flavors are of concern for many industries including food, health, and water. Metallic off-flavor, induced by ferrous sulfate solution (10mg/L), and its remediation using pre- and post-rinse treatments of water (control) or metal chelators, were studied. Metal chelators included lactoferrin (1 ?M), a natural metal-binding protein in milk and saliva, and EDTA (36 ?M), a synthetic chelator. Time-intensity (TI) evaluation (n=6, 4 female; age 40-70) of lingering metallic flavor indicated that metallic flavor decreased with a post-rinse adjuvant treatment of lactoferrin as indicated by a reduced maximum intensity and area under the curve compared to a pre-rinse treatment; EDTA and water post-rinses were equally effective for three of the TI parameters. Alterations in salivary components were studied in saliva collected (n=8; 5 female, age 40-70) after sipping a lactoferrin solution (1?M) followed with a ferrous sulfate sample (10 mg/ml) to stimulate metallic flavor, as compared to unstimulated whole saliva. Protein concentration, oral lipid oxidation as indicated by thiobarbituric acid reactive substances assay, and iron concentration were determined on individual saliva samples, with no significant differences found between treatments (p>0.05). Protein patterns were qualitatively characterized for each pre-rinse and metallic stimuli from four panelists by two-dimensional gel electrophoresis. A consistent pattern of regions containing major salivary components was observed. This research has shown that lactoferrin protein is a potential natural alternative to synthetic EDTA for reducing iron-induced metallic off-flavors. This study provides a foundation of method development to better understand salivary protein interaction with metals and flavor perception.Master of Science in Life Sciences
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Fulton, Benjamin L. "2D-PAGE Analysis of Myocardial Collagen in Male and Female Spontaneously Hypertensive Rats." Connect to resource online, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1219668882.
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Dwyer, Virginia Michelle Gregory 1955. "A STUDY OF PINEAL GLAND POLYPEPTIDES AND PROTEINS BY POLYACRYLAMIDE GEL ISOELECTRIC FOCUSING (PAG-IEF) AND TWO-DIMENSIONAL ELECTROPHORESIS (2DE) (BRAIN REGIONS)." Thesis, The University of Arizona, 1986. http://hdl.handle.net/10150/276560.
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Trein, Cristina Rodrigues. "Eletroforese bidimensional em gel de poliocrilamida do plasma seminal equino e a sua relação com a congelabilidade do sêmen." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2012. http://hdl.handle.net/10183/39292.
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O objetivo deste estudo foi avaliar o perfil proteico do plasma seminal equino utilizando eletroforese bidimensional de gel de acrilamida (2D-PAGE) e determinar se algumas das proteínas presentes estavam relacionadas com a congelabilidade do sêmen. O plasma seminal foi coletado de dez garanhões, de alta e baixa congelabilidade de sêmen, provenientes do Haras Estatal da Baixa Saxônia, na cidade de Celle, na Alemanha e rotineiramente utilizados em programas de inseminação artificial. Vinte e cinco bandas proteicas foram encontradas nos géis bidimensionais (12%) e sete delas foram identificadas por MALDI-MS. Das 25 proteínas encontradas nas amostras de plasma seminal dos garanhões, duas bandas proteicas apresentaram densidade óptica superior (P<0,05) nas amostras de garanhões de alta congelabilidade o sêmen: as bandas 5 (80-85 kDa, pI 7,54), que foi identificada como CRISP3 e a 45 (18,2 kDa, pI 5,0-5,2) identificada como HSP-2. Contrariamente a banda 7 (75,4 kDa, pI 6,9 – 7,4), identificada como lactoferrina, a 15 (26,7 kDa, pI 5,51) identificada como calicreína, a 25 (25 kDa, pI 7,54) como CRISP3 e a 35 (13,9 kDa, pI 3,8 – 4,2) que foi identificada como HSP-1, apresentaram valores de densidade óptica superior (P<0,05) nos reprodutores de baixa congelabilidade do sêmen. As proteínas foram identificadas através de espectometria de massa MALDI-MS. As evidências encontradas neste experimento mostram que existem diferenças no perfil proteico dos reprodutores de alta e baixa congelabilidade do sêmen, sugerindo as proteínas CRISP3 e a HSP-2 como possíveis marcadores da alta congelabilidade de sêmen de garanhões.The objective of this study was to evaluate protein profile of equine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and to find if any of these proteins were related to semen freezability. Seminal plasma was collected from 10 stallions of high and low semen freezability routinely used in AI programs from the State Stud of Lower Saxony and Artificial Insemination Center. Twenty five protein spots were identified from the 2 D gel (12%), seven of which were present in all samples. Of the 25 proteins found in the research stallions, two spots showed superior relative content (P<0.05) in seminal plasma samples collected from stallions with high semen freezability: the spots 5 (80-85 kDa, pI 7.54) was identified as CRISP3 and 45 (18.2 kDa, pI 5.0-5.2) was identified as HSP-2. Conversely, the spot 7 (75.4 kDa, pI 6.9 – 7.4) was identified as lactoferrin, 15 (26.7 kDa, pI 5.51) was identified as kallikrein, 25 (25 kDa, pI 7.54) was identified as CRISP3 and 35 (13.9 kDa, pI 3.8 – 4.2) was identified as HSP-1, showed superior relative protein content (P<0.05) on seminal plasma samples from stallions with low semen freezability. The proteins were identified by MALDI-MS. There were differences in the seminal plasma protein profile between stallions with high and low semen freezability. It may be suggested that CRISP3 and HSP-2 may be considered possible seminal plasma markers of high semen freezability.
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Misztal, David Richard Biotechnology & Biomolecular Sciences Faculty of Science UNSW. "Identification of cellular changes associated with increased production of human follicle stimulating hormone in a recombinant Chinese hamster ovary cell line." Publisher:University of New South Wales. Biotechnology & Biomolecular Sciences, 2008. http://handle.unsw.edu.au/1959.4/41326.
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A proteomics approach was used to identify proteins potentially implicated in the cellular response concurrent with elevated production levels of human follicle stimulating hormone in a recombinant Chinese hamster ovary cell line (Darren cells), using zinc and sodium butyrate in the production media to increase expression. To this end, 2-dimensional gel electrophoresis (2-DGE) was utilized. Firstly, several aspects of 2-DGE were developed for this investigation. Gel drying conditions were optimized, and a glycine-free blotting method is described which achieved greater efficiency in rapid transfer of proteins than those previously described. Next, hFSH expression was characterized in Darren cells. An ELISA developed for this investigation examined intracellular (expression) and extracellular (secretion) of hFSH during increased expression. These results show a disproportionate increase in intracellular hFSH (188%) expression above extracellular hFSH (41%).
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Ghafouri, Bijar. "Proteomics of the upper airways : studies on a new lipopolysaccharide-binding protein, PLUNC /." Linköping : Dept. of Molecular and Clinical Medicine, Linköping University, 2005. http://www.bibl.liu.se/liupubl/disp/disp2005/med927s.pdf.
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Silva, Fábio Arlindo. "Selênio em tilápia do Nilo utilizando eletroforese em gel e espectrometria atômica /." Botucatu : [s.n.], 2009. http://hdl.handle.net/11449/104086.
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Resumo: O presente trabalho teve como objetivo investigar a presença de selênio em spots protéicos de amostras de plasma, músculo e fígado de tilápia do Nilo (Oreochromis niloticus) obtidos após separação das proteínas por eletroforese em gel de poliacrilamida em segunda dimensão (2D-PAGE) e posterior avaliação qualitativa por fluorescência de raios-X com radiação síncrotron (SR-XRF). A análise dos espectros de fluorescência obtidos indicaram a presença de selênio em oito proteínas do plasma, seis proteínas do músculo e cinco proteínas do fígado. Observou-se que o selênio está distribuído em sua maioria em proteínas com massa molar menor que 50 kDa. Proteínas acima de 50 kDa foram encontradas somente no plasma.Abstract: An investigation was made into selenium in protein spots of samples of plasma, muscle and liver of Nile tilapia (Oreochromis niloticus) obtained after protein separation by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and subsequent qualitative evaluation by synchrotron radiation X-ray fluorescence (SR-XRF). An analysis of the fluorescence spectra indicated the presence of selenium in eight plasma proteins, six muscle proteins, and five liver proteins. Selenium was found to be distributed mainly in proteins with a molar mass smaller than 50 kDa. Proteins with a molar mass higher than 50 kDa was found only in the plasma.
Orientador: Pedro de Magalhães Padilha
Coorientador: Marco Aurélio Zezzi Arruda
Banca: Paulo Roberto Rdrigues Ramos
Banca: Ricardo de Oliveira Orsi
Banca: Gustavo Rocha de castro
Banca: Paulo dos Santos Roldan
Doutor
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Guterres, Sheila Barreto. "Busca de biomarcadores para esquizofrenia em plaquetas utilizando eletroforese diferencial em gel bidimensional (2D-DIGE) e espectrometria de massas." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/75/75132/tde-16112011-150931/.
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A esquizofrenia é uma doença crônica, grave e incapacitante que afeta cerca 24 milhões de pessoas em âmbito mundial. É caracterizada por uma desorganização no pensamento que prejudica a funcionalidade do indivíduo. Existem intervenções que são efetivas e contribuem para a diminuição da prevalência do transtorno, pois ajudam o portador a levar uma vida produtiva e integrada à sociedade, porém devem ser ministradas nos estágios iniciais da doença. No entanto, existe uma grande dificuldade em se diagnosticar a esquizofrenia precocemente devido a sua complexidade e às sutilezas dos seus sintomas apresentados antes do surgimento da psicose. O cérebro não é acessível a exames invasivos in vivo e por esse motivo a exploração de fluidos periféricos é de grande importância. As plaquetas e neurônios serotonérgicos possuem características bioquímicas e morfológicas em comum que possibilitam a comparação entre a estrutura e a função de ambos e, por causa dessa similaridade, muitos trabalhos utilizam plaquetas como modelo para o estudo de doenças neuropsiquiátricas, inclusive a esquizofrenia. A detecção precoce da esquizofrenia é um objeto de investigação atual e relevante não somente para revolucionar os meios atuais de diagnóstico, mas também para desenvolver novos tratamentos aplicados aos estágios iniciais da doença, diferenciar os subgrupos de doentes e monitorar as intervenções preventivas. A proposta do presente trabalho é fazer o estudo da expressão de proteínas em plaquetas de pacientes esquizofrênicos e controles com o objetivo de identificar proteínas candidatas a biomarcadores utilizando técnicas proteômicas quantitativas e confiáveis, como 2D-DIGE e a espectrometria de massas.Schizophrenia is a disabling, serious, and chronic illness, which affects about 24 million people worldwide. It is characterized by a severe disorganization of the thoughts that harms the social life of patients becoming them dependent of the family and/or government. There are effective treatments that contribute to decrease the prevalence of the disorder because they improve the life and social conditions of the patients, but they are only advantageous if the intervention is made in the early stages of the disease. It is difficult to obtain early diagnosis due to the complexity of the disease and its insidious symptoms before the beginning of the psychosis. The brain is not easily accessed in vivo and, because of this, it is very important to study the peripheral tissues like blood, which makes the use of the platelets very interesting. Furthermore, platelets and serotonergic neurons share biochemical and morphological characteristics that allows the comparison between structure and function of both. From these similarities many authors has used platelets as a neuron model to study many neurodegenerative diseases including schizophrenia. The early detection of schizophrenia is a current and suitable goal, not only to improve the early diagnosis but also to develop new treatments, differentiate the subtypes, and monitor the preventive interventions. The purpose of this project is to do a comparative screening of expressed proteins in platelets from schizophrenics and controls with the objective of finding differently expressed proteins that could be candidates to biomarkers using 2D-DIGE and mass spectrometry.
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Hadjem, Ammar Saïd. "Traitement d'image du gel d'électrophorèse bidimensionnelle." Nancy 1, 1988. http://www.theses.fr/1988NAN10143.
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L'objectif de ce travail est le traitement d'images du gel d'électrophorès bidimensionnelle (détection des taches de protéines, détermination automatique du nombre de taches de protéines sur le gel, la surface de chaque tache, leur position ainsi que leur densité optique ou en d'autres termes leur intensité lumineuse respective) en utilisant l'interface de numérisation associé à une caméra vidéo et à un micro-calculateur. Le travail a consisté, d'une part à mettre en œuvre une façon pratique cette nouvelle méthode d'investigation et d'autre part, à créer le logiciel assurant un fonctionnement jugé optimal pour ce type de fonctions (traitements d'images vidéo). L'autre partie de mon sujet est le travail expérimental en mettant au point les supports logiciels et techniques de la manipulation. En préliminaire est exposée une méthode originale de stockage des données numériques relatives à la partie comprise dans la fenêtre de mémorisation à position programmable ; la procédure de transfert de ces données dans le micro-calculateur en vue de leur traitement informatique est également présentée
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Nguyen-Lefebvre, Anh Thu. "Implication des protéines ribosomiques dans le processus de transformation induit par l’oncogène v-erbA." Thesis, Lyon 1, 2012. http://www.theses.fr/2012LYO10058/document.
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L’oncogène v-erbA transforme les progéniteurs érythrocytaires primaires aviaires (T2EC) en bloquantleur engagement d’un programme d’auto-renouvellement vers un programme de différenciation. Unecomparaison trancriptomique de T2EC exprimant soit v-erbA, soit une forme non transformante de verbAa été réalisée par SAGE et RT-qPCR. Seuls quelques uns, mais pas tous les messagers codant lesprotéines ribosomiques sont réprimés. Ces résultats suggèrent que v-erbA pourrait moduler lacomposition des ribosomes et/ou moduler les fonctions extra-ribosomiques de protéines ribosomiquesspécifiques. Ainsi, nous avons décidé d’analyser le taux des protéines ribosomiques associées auxribosomes par 2D-DIGE à partir des ribosomes purifiés. L’analyse statistique effectuée sur 4expériences indépendantes avec des marquages inversées a montré de manière significative que letaux de RPL11 est inférieur dans les T2EC exprimant v-erbA comparé à ceux exprimant la forme nontransformante de v-erbA. Ces données indiquent l’existence de ribosomes dépourvus de RPL11 dansles T2EC sous l’effet de v-erbA. Les résultats des expériences d’immunoprécipitation ont conforté cettehypothèse. L’ensemble des résultats obtenus suggèrent l’implication des protéines ribosomiques, etspécialement celle de RPL11, dans les processus de transformation induite par l’oncogène v-erbA, à lafois au niveau de la traduction, et probablement par sa fonction extra-ribosomique. L’analyse de lafonction biologique de RPL11 a montré qu’une sur-expression de RPL11 dans les T2EC retarderait laprolifération cellulaireThe v-erbA oncogene transforms chicken erythroid progenitors by blocking their differentiation andpreventing them to exit a state of self-renewal. The transcriptome of primary avian erythroidprogenitors cells (T2EC) expressing either v-erbA or a non-transforming form of v-erbA werecompared by SAGE. Only some, but not all, mRNAs encoding ribosomal proteins were shown to beaffected. These results suggest that v-erbA could modulate the composition of ribosomes and/ormodulate the extraribosomal functions of specific ribosomal proteins. We therefore decided to analyzethe level of ribosomal proteins associated to ribosomes by 2D-DIGE performed on purified ribosomes.A statistical analysis performed on 4 independent flip-flop experiments demonstrated that the level ofRPL11 is significantly lower in T2EC expressing v-erbA as compared to the non-transforming form ofv-erbA. These data suggest the presence of ribosomes without RPL11 in T2EC expressing v-ErbA.Results obtained from immunoprecipitation experiments were strengthened this hypothesis. The set ofthese data evoke the involvement of ribosomal proteins, and specially RPL11, in the v-erbAtransformation process both at the translational level and possibly in its extra-ribosomal function.Overexpression of RPL11 in T2EC showed a decrease of cell proliferation
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Chen, Zhaoyuan. "Development of Methods for the Study of Phosphoproteins." Diss., CLICK HERE for online access, 2006. http://contentdm.lib.byu.edu/ETD/image/etd1629.pdf.
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Chen, Kan. "Applications of Mass Spectrometry to Analysis of Prodiginines, Bioactivated Methylenedianiline Intermediates, and Hypoxia Induced Changes in the Zebrafish Skeletal Muscle Proteome." ScholarWorks@UNO, 2008. http://scholarworks.uno.edu/td/899.
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Mass spectrometry coupled with liquid chromatography and gel electrophoresis enables separation and detection of components in a complex mixture. During the last two decades, its applications were dramatically extended and remarkable progress has been made in many fields, in particular, environmental and biological analyses. This dissertation focuses on identification and characterization of biologically active compounds and comparative analysis of protein expression changes. The first two projects (Chapters 2 and 3) focus on the application of LC/MS approach to profile the bioactivated intermediates of 4, 4'-methylenedianiline (DAPM) from rat vascular smooth muscle cells (VSMCs) and bile. In our study, several DAPM metabolites were detected and characterized in detail by liquid chromatography-electrospray tandem mass spectrometry. The structural assignments of these metabolites from VSMCs and rat bile significantly improve our understanding of DAPM biotransformations and toxicity. The third project described in Chapter 4 focuses on using electrospray tandem mass spectrometry (ES-MS/MS) and theoretical calculation (GAUSSIAN 03 program) to investigate the unusual methyl radical loss and consecutive fragment ions that dominate the low-energy collision induced dissociation (CID) mass spectra of prodiginine compounds. Structures of the fragment ions are proposed and explanations are given to rationalize the observed competition between the formation of even-electron ions and radical ions. Our study shows that the lower apparent threshold associated with methyl radical loss points to a lower kinetic barrier. In Chapter 5, hypoxia-induced changes of zebrafish skeletal muscle were studied using two-dimensional difference in-gel electrophoresis (2D-DIGE) in vivo after 48 h in hypoxia vs. normoxia. The results showed that proteins involved in mitochondrial oxidative metabolism are down-regulated, whereas glycolytic enzymes are up-regulated to compensate for the loss of ATP synthesis in aerobic metabolism. The up-regulation of two spots identified as hemoglobin variants was also observed. These protein expression changes are consistent with a hypoxic response that enhances anaerobic metabolism or O2 transport to tissues.
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Daveau, Romain. "One-Dimensional Electrophoresis Gel Anlalysis Tool, ODEGAT. Elaboration d’un outil bio-informatique d’aide à la mise en évidence de marqueurs à visée diagnostique et pronostique : application à la polyarthrite rhumatoïde." Rouen, 2008. http://www.theses.fr/2008ROUES033.
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La polyarthrite rhumatoïde (PR) demeure aujourd’hui un vrai problème de santé publique. Rhumatisme inflammatoire chronique le plus fréquent de l’adulte, la PR est une affection autoimmune poly-factorielle aux manifestations cliniques initiales et au devenir variables. Responsable de destructions ostéo-cartilagineuses aux conséquences fonctionnelles graves, la PR se traduit généralement par une atteinte bilatérale et symétrique des petites et moyennes articulations, source de handicap pour les malades dont la prise en charge est fortement compliquée par l’absence de critères diagnostiques et pronostiques clairs. Pourtant des solutions thérapeutiques existent : anti-TNF-, IL-1 Ra, anti-CD20…D’autant plus efficaces qu’elles sont administrées tôt mais très coûteuses, ces « biothérapies » ciblées ne sont pour l’instant prescrites qu’en 2e intention, après l’échec de traitements de fond classiques sur l’activité de la maladie et l’atteinte structurale. Parce qu’elles continuent d’« échapper » aux différents indicateurs clinico-biologiques à ce jour disponibles, ces 10 – 15% de PR, dites sévères et requérant de telles molécules, doivent donc en priorité être identifiées avant que n’apparaissent d’irréversibles lésions. Adossés à une cohorte de rhumatismes inflammatoires débutants (VErA) et à l’utilisation de langages informatiques (Perl, R) appropriés, ces travaux s’intéressent aux questions essentielles du diagnostic et du pronostic précoce. En terme de diagnostic, la contribution des facteurs génétiques (HLA-DR1, TNFRII*196R et PTPN22*1858) se révéla limitée comparativement aux auto-anticorps (Ac) qui restent les marqueurs les plus pertinents. S’agissant du pronostic, une étude « pilote » menée sur des biopsies synoviales démontrait l’intérêt du CD20 tissulaire dans un modèle de régression linéaire associant facteurs rhumatoïdes, anticorps anti-protéines citrullinées (ACPA) et RANKL (marqueur du remodelage osseux), prédictif de l’atteinte articulaire à 3 ans dans un panel de 14 patients. En raison de l’insuffisance de marqueurs diagnostiques et pronostiques, la recherche de nouvelles cibles immunologiques dans le cadre du programme ACPRA nous amena à la conception d’un outil bioinformatique d’analyse de gels électrophorétiques-1D baptisé ODEGAT. Appliqué aux immuno-empreintes de 110 sérums de patients de la cohorte VErA, ODEGAT contribua à l’identification de 4 auto-Ac originaux : anti-PGK1, -STIP1, -FUSE-BP 1 et 2, présents pour au moins l’un d’entre eux dans 40% des PR initialement négatives pour les ACPA. Enfin, des sérums de 10 témoins non arthritiques et 21 malades divisés en 11 PR sévères et 10 bénignes, ODEGAT mettait en évidence 5 bandes polypeptidiques dont les niveaux relatifs d’expression permettaient d’appréhender l’impact radiologique à court terme avec une valeur prédictive négative (resp. Positive) de 90% (resp. 82%). Perspective de cette thèse et objectif ultime du projet APOTRA, ce dernier résultat « prometteur » est à l’origine de la réalisation prochaine d’une « biopuce » à protéines qui devrait être testée sur un échantillon plus important de PR : la cohorte nationale ESPOIRRheumatoid arthritis (RA) still represents a serious challenge in terms of public health. Remaining the most frequent chronic inflammatory rheumatism, RA is an auto-immune disease with multiple origins. Moreover, its clinical presentation as well as its evolution are heterogeneous. RA generally consists in a bilateral and symmetrical inflammation of small and intermediate joints, responsible for severe bone destruction and leading to heavy functional consequences and disability for patients. Patient management is also highly complicated by the absence of clear criteria for diagnosis and prognosis of the disease. Yet, therapeutic solutions exist : anti-TNF-, IL-1 Ra, anti-CD20. . . These costly specific drugs are more efficient when administered early, but are often used as a second line of treatment after classical therapies have failed. 10 – 15% of RA are considered as severe and require such treatments. They thus need to be identified early to insure optimal management and prevent the occurence of irreversible lesions. However, none of the existing clinical or biological indicators commonly used allow to distinguish such severe forms of RA. This work focuses on the main questions of early diagnosis and prognosis of RA based on a cohort of patients with early inflammatory rheumatisms (VErA) and using appropriate computer languages (Perl, R). In terms of diagnosis, contribution of genetic factors (HLA-DR1, TNFRII*196R et PTPN22*1858T) turned out to be limited relatively to auto-antibodies which remain the most pertinent markers. As for prognosis, a pilot study performed on synovial biopsies demonstrated the role of tissue CD20 levels as predictive of articular lesions at 3 years in a panel of 14 patients from a linear regression model associating rheumatoid factors, anti-citrullinated protein antibodies (ACPA) and RANKL (bone remodelling marker). Confronted to the insufficiency in diagnostic and prognostic markers, search for new immunological targets as part of the ACPRA program led us to develop a new bioinformatics tool dedicated to 1-D electrophoretic gels named ODEGAT. 110 immunoblots of patient sera from the VErA cohort were tested and ODEGAT contributed to the identification of 4 original autoantibodies : anti-PGK1, -STIP1, -FUSE-BP 1 et 2. At least one of these markers was present in 40% of RA initially tested negative for ACPA. Finally, in a comparison of 10 healthy controls and 21 patients divided in 11 severe RA and 10 benign, ODEGAT revealed 5 polypeptidic bands with relative expression levels correlated to short term radiologic impact with a negative (resp. Positive) predictive value of 90% (resp. 82%). As a perspective of this PhD work and as main objective of the APOPTRA project, this last promising result will soon permit the development of a protein array which will be tested on a large number of RA, from the national ESPOIR cohort
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Sayahtaheri, Sousan. "Two-Dimensional Gel Electrophoresis of in Vivo and in Vitro Synthesized Proteins, Antigenic Proteins, and Cross-Reactive Antigens in Treponema Pallidum Subsp. Pallidum Nichols Strain and Treponema Phagedenis Biotype Reiter." Thesis, University of North Texas, 1989. https://digital.library.unt.edu/ark:/67531/metadc935721/.
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Two-dimensional electrophoretic protein profiles of in vivo and in vitro propagated T.pallidum subsps. pallidum Nichols strain were analyzed and compared. This comparative analysis revealed two in vitro synthesized, cytoplasmic cylinder-associated polypeptides with molecular masses 29.5 and 34.7 kDa, pI 5.62, and one in vitro "lost" polypeptide with molecular mass 34.7 kDa, pI 5.34. integral membrane proteins of in vitro and in vivo propagated T. pallidum was identified by phase partitioning with the nonionic Triton X-114, and twelve outer membrane-associated, antigenic proteins were identified in western blots probed with pooled human secondary syphilitic sera. The solubilization of the outer membrane of T. pallidum with Triton X-114 were monitored by electron microscopy. Treatment of freshly harvested 35S labeled T. pallidum with 1% Triton X-114 resulted in solubilization of the outer membrane and reduction of the diameter of the treponemes from .14 +/- .02 micrometers to .095 +/- .003 micrometers. Examination of thin sections of untreated organisms showed integrity of outer and cytoplasmic membranes. In contrast, thin sections of Triton X-114-treated trponemes showed integrity of the cytoplasmic membrane but the loss of the outer membrane. The cytoplasmic cylinders generated by detergent treatment retained their periplasmic flagella, as judged by electron microscopy and immunoblotting. Integral membrane proteins of Treponema phagedenis were also identified by phase partitioning with Triton X-114, and sizteen cross-reactive, outer membrane-associated, outer membrane-associated, antigenic polypeptides were identified in western blots probed with pooled human secondary syphilitic sera. The results of this study indicate that tow-dimensional protein profiles of in vivo and in vitro propagated T.pallidum are almost identical except for the differences mentioned. This results also indicate that 1% Triton X-114 selectively solubilizes the outer membrane, and the antigenic hydrophobic proteins present in the detergent phrase are located exclusively in the outer membrane.
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Hirschberg, Daniel. "Sample preparation and mass spectrometry in proteome studies /." Stockholm, 2004. http://diss.kib.ki.se/2004/91-7349-934-x/.
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Coaker, Gitta Laurel. "Genetic and biochemical characterization of resistance to bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis." The Ohio State University, 2003. http://rave.ohiolink.edu/etdc/view?acc_num=osu1069188955.
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Qiu, Linghua. "Differentially Expressed Proteins in the Pancreas of Diabetic Mice." Ohio University / OhioLINK, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1125866599.
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Grinyer, Jasmine. "Proteomic analysis of the biological control fungus Trichoderma." Doctoral thesis, Australia : Macquarie University, 2007. http://hdl.handle.net/1959.14/12407.
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Thesis by publication."August 2006"
Thesis (PhD)--Macquarie University, Division of Environmental & Life Sciences, Dept. of Biological Sciences & Dept. of Chemistry & Biomolecular Sciences), 2007.
Bibliography: leaves 157-183.
1. Introduction -- 1.1. Proteomics and two-dimensional electrophoresis -- 1.2. A proteomic approach to study the filamentous fungus Trichoderma -- 1.3. Aims of the thesis -- 2. Materials and methods -- 3. Results and discussion -- 3.1. Method development for the display and identification of fungal proteins by 2DE and mass spectrometry -- 3.2. Discovery of novel determinants in the biological control of phytopathogens by Trichoderma atroviride -- 3.3. Summary and concluding remarks.
Trichoderma harzianum and T. atroviride are filamentous fungi commonly found in soil. Both display biocontrol capabilities against a range of phytopathogenic fungi including Rhizoctonia solani and Botrytis cinerea which are known pests of hundreds of commercially important crops including tomatoes, potatoes, beans, cucumber, strawberries, cotton and grapes. These Trichoderma species secrete a combination of enzymes degrading cell walls and antibiotics to overgrow and kill fungal phytopathogens. They are seen as an environmentally friendly alternative to chemical fungicides currengly used on crops.
A proteomic approach was taken to separate and identify proteins from a strain of T. harzianum with well established biocontrol properties. Several methods were developed in this thesis to display the whole proteome content and several subcellular proteome fractions from T. harzianum. Proteins were separated by two-dimensional electrophoresis and identified by mass spectrometric methods. The resulting proteomic maps represent the first extensive array of cellular and sub-cellular proteomes for T. harzianum.
Cellular protein patterns of T. atroviride (T. harzianum P1) grown on media containing either glucose or R. solani cell walls were compared by differential gel electrophoresis to identify a suite of new proteins involved in the biological control response. Twenty four T. atroviride protein spots up-regulated in the presence of the R. solani cell walls were identified by mass spectrometry and N-terminal sequencing. Proteins identified from this study included previously implicated enzymes degrading cell walls and three novel proteases, vacuolar serine protease, vacuolar protease A and trypsin-like protease. The genes encoding two of these proteases, vacuolar protease A and vacuolar serine protease have been cloned by degenerate primer PCR and genomic walking PCR and sequenced. The gene sequences and protein sequences derived from these genes have been partially characterised.
Mode of access: World Wide Web.
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Paspuleti, Sreelatha. "Isolation and Identification of O-linked-β-N-acetylglucosamine Modified Proteins (O-GlcNAc) in the Developing Xenopus laevis Oocyte." Scholar Commons, 2004. https://scholarcommons.usf.edu/etd/809.
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Oocyte development in Xenopus laevis spans six morphologically distinct stages (stage I-VI), and is associated with a decrease in protein O-GlcNAc levels. As a first step in elucidating the role of O-GlcNAc in developing oocytes, initial efforts were focused on isolation and identification of fifteen modified proteins that decrease during oocyte development. Stage I oocytes due to their high amounts of these proteins, were used as starting material for purification. Multiple affinity and specific antibody based purification technique were initially used in an attempt to enrich the O-GlcNAc proteins. Due to the unique properties of the proteins ultimately identified, these techniques were unable to provide sufficient material for sequencing. However, differential centrifugation coupled with 2D-gel electrophoresis was highly successful. The majority of isolated proteins were strongly basic in nature with pIs 8-10. Coomassie stained bands from 2D-analysis were trypsin digested, and peptides were sequenced by mass spectroscopy (Finnigan LCQ). Mass data were interpreted by Bioworks software, and protein sequences were compared to multiple protein databases. Initially, six proteins were identified as Thesaurin a (42Sp50), cytoplasmic mRNA binding protein p54, y-box homolog, Xp 54 (ATP dependent RNA helicase p54), Vg1 RNA binding protein variant A, Zygote arrest 1(Zar1) and Poly (A) binding protein (PABP). Thesaurin a, the main component of 42S particle of previtellogenic oocytes (stages I-III) is involved in tRNA storage and possess low tRNA transfer activity; y-box factor homolog and Xp54 are present in oocyte mRNA storage ribonucleoprotein particles; Vg1 RBP variant A associates mVg1 RNA to microtubules in order to translocate to the vegetal cortex; Zar1 is involved in oocyte-to-embryo transition; and PABP initiates mRNA translation. This study is the first to characterize these oocyte specific proteins as O-GlcNAc modified proteins. Overall, the presence of several O-GlcNAc proteins in oocytes, the reduction in their levels/ O-GlcNAc levels, and the variation in maturation time in the presence of HBP-flux modulators in developing oocyte indicates O-GlcNAc may play important roles in metabolism, cell growth and cell division of X. laevis oocytes. Therefore, identifying the remainder of these proteins and elucidating the O-GlcNAc role in their function is a worthwhile pursuit.
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Abbaraju, Naga Vijayalaxmi. "Patterns of protein expression in tissues of the killifish, Fundulus heteroclitus and Fundulus grandis." ScholarWorks@UNO, 2011. http://scholarworks.uno.edu/td/113.
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Fundulus is a diverse and widespread genus of small teleost fish of North America. Due to its high tolerance for physiochemical variation (e.g. temperature, oxygen, salinity), Fundulus is a model organism to study physiological and molecular adaptations to environmental stress. The thesis focuses on patterns of protein expression in Fundulus heteroclitus and F. grandis.The patterns of protein expression were investigated using traditional methods of enzyme activity measurements and recent proteomic approaches. The findings of the study can be used to guide future studies on the proteomic responses of vertebrates to environmental stress. Chapter 2 focuses on measurement of the temporal effects of oxygen treatments on the maximal specific activities of nine glycolytic enzymes in liver and skeletal muscle during chronic exposure (28d) of Fundulus heteroclitus. The fish was exposed to four different oxygen treatments: hyperoxia, normoxia, moderate hypoxia, and severe hypoxia. The time course of changes in maximal glycolytic enzyme specific activities was assessed at 0, 8, 14 and 28 d. The results demonstrate that chronic hypoxia alters the capacity for carbohydrate metabolism in F. heteroclitus, with the important observation that the responses are both tissue- and enzyme-specific. Chapter 3 studies the effect of tissue storage on protein profile of tissues of F. grandis. The technique of one dimensional gel electrophoresis (1D-SDS-PAGE) was used to assess the effects of tissue sampling, flash frozen in liquid nitrogen versus immersion of fresh tissue in RNA later, for five tissues, liver, skeletal muscle, brain, gill, and heart, followed by LC-MS/MS to identify protein bands that were differentially stabilized in gill and liver. The study shows that, in F. grandis, the preferred method of preservation was tissue specific. xi Chapter 4 focuses on the use of advanced 2DE-MS/MS to characterize the proteome of multiple tissues in F. grandis. Database searching resulted in the identification of 253 non-redundant proteins in five tissues: liver, muscle, brain, gill, and heart. Identifications include enzymes of energy metabolism, heat shock proteins, and structural proteins. The protein identification rate was approximately 50 % of the protein spots analyzed. This identification rate for a species without a sequenced genome demonstrates the utility of F. grandis as a model organism for environmental proteomic studies in vertebrates.
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Apraiz, Larrucea Itxaso. "Development and application of a proteomic approach to the assessment of pollution in the marine environment." Doctoral thesis, Stockholms universitet, Institutionen för biokemi och biofysik, 2009. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-26150.
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Today, assessment of the health of coastal waters is recognized as being important for both the conservation of nature and well-being of humans. Anthropogenic pollution has been the focus of extensive research for some time and a variety of programs for the monitoring and assessment of environmental pollution have been developed. Determination of the levels of pollution in sensitive ‘sentinels’ such as mussels, allows monitoring of these levels in a given area over a prolonged period of time. Furthermore, the biological effects of pollution are reflected in a series of biomarkers, none of which provides a general picture of the sentinel’s state of health and all of which are individually specific for certain pollutants and influenced by both biotic and abiotic factors. In an attempt to improve biomonitoring of marine pollution, we have developed two proteomic approaches here. In the first portion of the thesis, a proteomic analysis was performed on peroxisomes isolated from mussels exposed either to one of three model anthropogenic pollutants, or two different types of crude oil, or from mussels exposed to the Prestige oil spill. Application of two-dimensional electrophoresis (2-DE) provided protein expression signatures (PES) for exposure to these different pollutants.Furthermore, several individual protein components of these PES could be putatively identified. In the second portion of this work, such analysis of subproteomes was developed further in order to improve the applicability of this approach to biomonitoring. A simple fractionation procedure in combination with liquid chromatography and 2-DE provided samples from mussels residing in different regions of a pollution gradient around the harbor of Gothenburg, as well as from mussels exposed to two types of fuel oil similar to that of the Prestige that were suitable for environmental proteomics. In addition, we constructed a model for this approach that can be cross-validated in the future and applied to assess sources of fuel oil pollution in connection with biomonitoring programs.
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Amelina, Hanna. "Proteomics in biomarker research : Insights into the effects of aging and environment on biological systems." Doctoral thesis, Stockholms universitet, Institutionen för biokemi och biofysik, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-56483.
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Proteomics is the global analysis of proteins that covers a broad range of technologies aimed at determining the identity and quantity of proteins expressed in the cell, their three-dimensional structure and interaction partners. In contrast to genome, proteome reflects more accurately on the dynamic state of the cell, tissue, or an organism. Therefore much is expected from proteomics to yield better disease markers for early diagnosis and therapy monitoring, as well as biomarkers that would indicate environmental exposure or provide prediction of biological age. In this thesis, I have developed and applied robust and sensitive subproteomic approaches to study the effect of aging as well as and environmental pollution using different animal models. In the first part, a high-throughput proteomic method based on liquid chromatography coupled to 2-dimensional gel electrophoresis (LC/2-DE) was developed. The usefulness of this method has been demonstrated by applying it to the assessment of marine pollution in a field experiment. Next, I have utilized this subproteomic approach to study the effect of aging in mouse kidney of both genders. As a result, a protein expression signature of aging kidney was obtained, revealing gender-dependent alterations in proteome profiles of aging mouse kidney. In order to further reduce the dynamic range of protein expression and increase the sensitivity of proteomic analysis, I have applied a shotgun mass spectrometry-based proteomic approach using isobaric tags for relative and absolute quantification (iTRAQ) coupled to liquid chromatography and tandem mass spectrometry (LC-MS/MS) to study age-related differences in peroxisome-enriched fractions from mouse liver. Only eight proteins showed statistically significant difference in expression (p<0.05) with moderate folds. This study indicates that age-depended changes in the liver proteome are minimal, suggesting that its proteome is efficiently maintained until certain age. Finally, in the context of aging studies and the role of peroxisomes in aging, I have tested the utility of cell-penetrating peptides (CPPs) as agents for protein delivery into acatalasemic peroxisomes using yeast as a model. The results obtained suggest that CPPs may be suitable for the delivery of antioxidants to peroxisomes and in future could provide a tool for the protein therapy of age-related diseases.At the time of the doctoral defense, the following publications were unpublished and had a status as follows: Paper 3: Submitted, Paper 4: Submitted.
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Martins, Carlo de Oliveira. "Análise proteômica diferencial em válvula mitral na doença reumática cardíaca." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/5/5146/tde-02082013-142739/.
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A Doença Reumática Cardíaca (DRC) é uma séria complicação de orofaringite causada por determinados sorotipos de Streptococcus pyogenes não tratada adequadamente em indivíduos suscetíveis. É um grande problema de saúde pública, principalmente nos países não desenvolvidos e em desenvolvimento, como Brasil, Índia, países da África, regiões de população aborígine da Austrália, e Egito. É altamente debilitante e com alta taxa de mortalidade devido ao comprometimento cardíaco. As lesões miocárdicas iniciais regridem, mas as lesões valvares, principalmente a mitral e a aórtica, são irreversíveis e progressivas. Muitos estudos já caracterizaram a resposta imune celular (linfócitos T) e humoral nos indivíduos acometidos pela doença. Mimetismo molecular e espalhamento de epítopo são os principais mecanismos que se pensa estar envolvidos na patogênese da DRC. Avaliamos, nesta pesquisa, o perfil de expressão proteica em valvas mitrais de indivíduos acometidos por DRC. Para detectar alterações específicas desta doença, comparamos as expressões de proteínas nos grupos portadores de DRC com insuficiência (DRC-INS) e com estenose (DRC-EST) a um grupo de indivíduos com degeneração mixomatosa de valva mitral (DMX) e outro sem valvulopatias (CTL). Alterações especificamente observadas em tecido mitral na DRC-INS ou DRC-EST em fases avançadas da doença podem explicar o mecanismo de desenvolvimento desses dois tipos de lesão. Foram encontradas 25 \"spots\", correpondendo a 29 proteínas diferencialmente expressas nos grupos com valvulopatias, refletindo principalmente alterações na matriz extracelular. Encontramos importante clivagem diferencial da vimentina, cuja proteína íntegra possui 54 kDa, formando fragmentos com ~40 e ~45 kDa, aumentados na DRC, principalmente na DRC-INS. O colágeno do tipo VI, com aproximadamente 95 kDa, encontrou-se com expressão diminuída exclusivamente no grupo DRC-INS. A Vitronectina foi encontrou-se aumentada em na DMX e na DRC-EST, em relação ao grupo controle, principalmente na DRC-EST. Lumican, por sua vez, teve expressão diminuída na DMX e na DRC-EST, apesar de possuir um único \"spot\" com expressão aumentada na DRC. Utilizando métodos de análise de padrões de expressão protéica in silico foram identificados conjuntos de proteínas capazes de discriminar as amostras de valva mitral por etiologia da doença. O presente trabalho pode auxiliar na elucidação dos mecanismos de desenvolvimento da doença e de alterações estruturais do tecido mitral em resposta às lesões autoimunes, bem como no diagnósticoda DRC.Rheumatic Heart Disease (RHD) is a serious complication of oropharingitis caused by some serotypes of Streptococcus pyogenes not properly treated in susceptible individuals. It is a public health concern, mainly for undeveloped and developing countries, such as Brazil, India, some countries in Africa, aboriginal regions in Australia, and Egypt. It is highly debilitating with a high mortality rate due to cardiac commitment. Initial myocardial lesions disappear, but valvar lesions, mainly mitral and aortic, are irreversible and progressive. Many studies have characterized cellular (T lymphocytes) and humoral responses in individuals affected by the disease. Molecular mimicry and epitope spreading are the main mechanisms thought to be involved in the pathogenesis of RHD. We evaluated, in this research, the profile of protein expression in mitral valves from individuals affected by RHD. To detect alterations specific of this disease, we compared protein expression in the group of RHD with regurgitation (RHD-RGT) and stenosis (RHD-STN) to a group of individuals with mitral valve myxomatous degeneration (MXD) and another group without valvulopathies (CTL). Alterations specifically observed in the mitral tissue of RHD-RGT and RHD-STN in advanced stages of the disease can explain the mechanism of development for these two kinds of lesions. Twenty-five spots, corresponding to 29 proteins were found to be differentially expressed in the valvulopathy groups, reflecting mainly alterations in extracellular matrix. We found important differential cleavage of vimentin, the whole protein having 54 kDa, in fragments with ~40 and ~45 kDa, increased in RHD, mainly in RHD-RGT. Collagen type-VI, with approximatelly 95 kDa, was found to have decreased expression exclusivelly in the RHD-RGT group. Increased expression of Vitronectin was detected in DMX and RHD-EST groups, compared to the CTL group, mainly in the RHD-STN. Lumican, in turn, had decreased expression in the MXD and RHD-STN groups. By using in silico methods for analysis of patterns of protein expression, we identified sets of proteins capable of discriminating mitral valve samples by disease etiology. The present study might help elucidating the mechanisms of disease development and structural alterations in the mitral tissue in response to the autoimmune lesions, as well as in the diagnosis of RHD.
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Rosengren, Åsa. "Cell-protein-material Interactions on Bioceramics and Model Surfaces." Doctoral thesis, Uppsala University, Surface Biotechnology, 2004. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-4688.
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The objective of this thesis was to investigate and characterize the interaction between blood proteins and different surfaces with emphasis on protein adsorption to bioceramics and model surfaces. Special effort was made to monitor the spontaneous and selective adsorption of proteins from human plasma and to examine the orientation, conformation and functional behavior of single proteins after adsorption.
Five different ceramic biomaterials: alumina (Al2O3), zirconia (ZrO2), hydroxyapatite (Ca10(PO4)6(OH)2) and two glass-ceramics, AP40 (SiO2-CaO-Na2O-P2O5-MgO-K2O-CaF2) and RKKP (AP40 with Ta2O3-La2O3), were exposed to human plasma and their protein binding capacities and affinities for specific proteins were studied by chromatography, protein assays, two-dimensional gel electrophoresis and Western blotting. The studies showed that all materials adsorbed approximately the same high amount of plasma proteins and that they therefore should be fully covered by proteins in an in vivo setting. The adsorbed proteins were different for most materials which could explain their previously observed different levels of tissue integration in vivo.
Four of the proteins that behaved differently, ceruloplasmin, prothrombin, α2-HS-glycoprotein and α1-antichymotrypsin, were selected for characterization with atomic force microscopy and ellipsometry. The studies, which were performed on ultraflat silicon wafers (silica), showed that the proteins oriented themselves with their long axis parallel to the surface or as in case of ceruloplasmin with one of its larger sides towards the surface. All of them had globular shapes but other conformational details were not resolved. Furthermore, prothrombin (none of the others) formed multilayers at high proteins concentrations.
The functional behaviour of the adsorbed proteins, referring to their cell binding and cell spreading capacity on silica and a positive cell adhesion reference surface (Thermanox®), was affected by the underlying substrate. Ceruloplasmin, α2-HS-glycoprotein and α1-antichymotrypsin stimulated cell attachment to silica, but suppressed attachment to Thermanox®. Prothrombin stimulated cell attachment to both surfaces. The attachment was in most cases mediated both by cell membrane-receptors (integrins) and by non-specific interactions between the cell and the material.
This thesis showed that the compositional mixture, orientation, conformation and functional behavior of the adsorbed proteins are determined by the properties of the underlying surface and if these parameters are controlled very different cellular responses can be induced.
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Kierul, Kinga. "Comprehensive proteomic study of Bacillus amyloliquefaciens strain FZB42 and its response to plant root exudates." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2013. http://dx.doi.org/10.18452/16805.
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Bacillus amyloliquefaciens FZB42 ist ein frei lebendes Bakterium, das Pflanzenwurzeln besiedelt und das Pflanzenwachstum durch viele verschiedene Wirkmechanismen anregt. In dieser Arbeit wurden die molekularen Grundlagen dieser positiven Wirkungen, die dieses „Pflanzenwachstum fördernde Rhizobakterium“ (PGPR) auf seine Wirte ausübt, untersucht. Um den gegenseitigen Austausch von B. amyloliquefaciens und seinen Wirtspflanzen zu entschlüsseln, wurden umfangreiche Proteomstudien durchgeführt. Es wurden Referenzkarten der extrazellulären und zytosolischen Proteinfraktionen erstellt. Die größte Anzahl an ausgeschiedenen Proteinen konnte während der stationären Phase beobachtet werden. Die identifizierten extrazellulären Proteine gehören verschiedenen Funktionsklassen an, wobei die prominentesten Klassen am Kohlenhydrat-Abbau und den Transport von Molekülen durch die Zellwand beteiligt sind. Die zytosolischen Extrakte von Kulturen, die in 1C-Medium bzw. Mineralmedium angezogen wurden, und in der zweidimensionalen Gelelektrophorese (2 DE) aufgetrennt wurden, ergaben 461 und 245 verschiedene Protein-Einträge. Die erstellten Referenz-Karten wurden anschließend verwendet, um Proteine und Prozesse, in an der Interaktion mit Pflanzen beteiligt sind, zu identifizieren. Dafür wurden die Bakterien Wurzelexudaten von Mais (Zea mays L.) ausgesetzt. Die Proteine aus zwei Stämmen, denen die globalen Transkriptionsregulatoren (Degu, AbrB) und vier Sigma-Faktoren (SigB, SigM, SigV, und SigX) fehlen, wurden ebenfalls untersucht, um ihre Beteiligung an den bakteriellen Reaktionen auf die Wurzelausscheidungen zu analysieren. Zusammenfassend ist dies die erste Studie, die umfangreiche Proteomdaten von Gram-positiven PGPR präsentiert, wobei gleichzeitig die Veränderung der Expression von extrazellulären und zytoplasmatischen Proteinen, nach Zugabe von Wurzelexudaten, ausgewertet wurde.Bacillus amyloliquefaciens strain FZB42 is a free-living bacterium that competitively colonizes plant roots and stimulates plant growth by many different modes of action. The molecular basis of singular beneficial effects that this Plant Growth-Promoting Rhizobacteria (PGPR) exert on their hosts have been studied. To decipher the molecular cross-talk of B. amyloliquefaciens and its’ host plants as a whole system, an extensive proteomic approach was performed. Reference maps of the extracellular and cytosolic protein fractions were established. The highest number of secreted proteins was observed during stationary growth phase. Identified extracellular proteins belong to different functional classes, with the most prominent classes involved in carbohydrate degradation and transportation of molecules across the cell wall. Cytosolic extracts obtained from cultures grown in 1C and minimal media subjected to the 2 Dimensional Electrophoresis (2 DE), revealed 461 and 245 different protein entries, respectively. Created reference maps were subsequently used to identify proteins and processes involved in the interaction with plants, prior to exposure of bacteria to maize (Zea mays L.) root exudates. The proteomics of two strains lacking expression of genes coding for global transcriptional regulators (degU, abrB) and four sigma factors (sigB, sigM, sigV, and sigX) were also inves-tigated, in order to analyse their involvement in bacterial responses to root exudates. In summary, this is the first study presenting comprehensive proteomics of Gram-positive PGPR, evaluating at the same time changes in protein expression caused by addition of root exudates at the extracellular and cytosolic level.
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YU, LONG XI. "Arns poly(a**(+)) cytoplasmiques specifiques de l'organogenese reproductrice a caractere sterile chez mercurialis annua l. (2n=16) : traduction in vitro-electrophorese bidimensionnelles, hybridations homologues et heterologues." Orléans, 1987. http://www.theses.fr/1987ORLE2050.
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Vileigas, Danielle Fernandes. "Proteoma miocárdico de ratos obesos por dieta Ocidental com disfunção cardíaca." Botucatu, 2019. http://hdl.handle.net/11449/181435.
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Orientador: Antonio Carlos CicognaResumo: A obesidade é uma doença metabólica complexa considerada uma pandemia global e associada à alta incidência de doença cardiovascular. O excesso de tecido adiposo pode promover mal adaptação que resulta em alterações na estrutura e função do coração; no entanto, os mecanismos não estão totalmente elucidados. A proteômica pode fornecer uma compreensão mais profunda do processo fisiopatológico e contribuir para a identificação de novos potenciais alvos terapêuticos. Portanto, o objetivo deste estudo foi avaliar a expressão proteica miocárdica em ratos saudáveis e obesos por dieta Ocidental, empregando duas abordagens proteômicas, para melhor compreender a rede de mecanismos inerentes à disfunção cardíaca na obesidade. Ratos Wistar foram distribuídos em dois grupos: controle (C, n = 13; dieta controle) e obeso (Ob, n = 13; dieta Ocidental) alimentados por 41 semanas. A obesidade foi determinada pelo índice de adiposidade. A função cardíaca foi avaliada pelo ecocardiograma e análise do músculo papilar isolado. A proteômica foi baseada em eletroforese em gel bidimensional (2-DE) juntamente com espectrometria de massa (LC-MS/MS) e cromatografia-líquida em nanofluxo com espectrometria de massa em tandem (nanoLC-MS/MS) seguida de quantificação label-free. Ratos obesos apresentaram aumento do índice de adiposidade e disfunção cardíaca sistólica e diastólica comparados aos controles. Um total de 82 proteínas miocárdicas foram identificadas como diferencialmente expressas entre os grupos ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Obesity is a complex metabolic disease considered a global pandemic and associated with high incidence of cardiovascular disease. The excess of adipose tissue may promotes maladaptation that result in alterations in structure and function of the heart; however, the mechanisms are not fully elucidated. Proteomics may provide a deeper understanding into the pathophysiological process and contribute to the identification of new potential therapeutic targets. Thus, the aim of this was evaluate the myocardial protein expression in healthy and obese rats, employing two proteomic approaches to better comprehend the network of mechanisms inherent to cardiac dysfunction in obesity. Male Wistar rats were distributed into two groups: control (C, n=13; standard diet) and obese (Ob, n=13; Western diet) fed for 41 weeks. The obesity was determined by adipose index. Cardiac function was evaluated by echocardiogram and isolated papillary muscle analysis. The proteomics was based on two-dimensional gel electrophoresis (2-DE) along with mass spectrometry identification (LC-MS/MS) and nano-liquid chromatography with tandem mass spectrometry (nanoLC-MS/MS) followed by label-free quantification. Obese rats showed increased adiposity index and systolic and diastolic cardiac dysfunction. A total of 82 myocardial proteins was identified as differentially expressed between C and Ob groups using two proteomic strategies, being 43 up- and 39 down-regulated by obesity. These proteins are involved in imp... (Complete abstract click electronic access below)
Doutor
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Guerrero, Barrado Pedro Enrique. "Altered glycosylation in pancreatic cancer: development of new tumor markers and therapeutic strategies." Doctoral thesis, Universitat de Girona, 2020. http://hdl.handle.net/10803/671007.
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Pancreatic Ductal Adenocarcinoma (PDA), the most common pancreatic cancer, remains one of the most lethal tumor types, with extremely low survival rates due to late diagnosis and resistance to standard therapies. Expression of tumor-associated carbohydrate antigens such as SLeX enhances several tumor features like cell migration or metastasis, representing a good source of new therapeutic targets and biomarkers. In this work, it has been studied the effect of the inhibition of the expression of the main genes that code for the enzymes responsible for the final steps of SLeX biosynthesis (the α2,3-sialyltransferases). Silencing of ST3GAL3 and ST3GAL4 in two PDA cell lines led to a significant reduction in SLeX associated with a decrease in cell migration, invasion and adhesion to E-selectin, decreasing PDA metastatic potential. Also, using glycoproteomic techniques, we identified the glycoprotein MFAP4 carrying SLeX as potential PDA biomarker since it is only present in PDA tissue samplesEl adenocarcinoma ductal pancreático o PDA, es el tipo más frecuente de cáncer de páncreas. El PDA es uno de los tumores más letales, con tasas de supervivencia extremadamente bajas, debido principalmente al diagnóstico tardío y su amplia resistencia a las terapias actuales. La expresión de antígenos carbohidratos asociados a tumores como el SLeX potencia varias características tumorales como la migración celular o metástasis, por lo cual representa una buena fuente de nuevas dianas terapéuticas y biomarcadores. En este trabajo, se ha estudiado el efecto de la inhibición de la expresión de los principales genes que codifican para las enzimas responsables de la biosíntesis de SLeX en sus etapas finales (las α2,3-sialiltransferasas). El silenciamiento de ST3GAL3 y ST3GAL4 en dos líneas celulares de PDA produjo a una reducción significativa en los niveles de SLeX. Este descenso se asoció con una disminución en la migración, invasión y adhesión celular a E-selectina, disminuyendo el potencial metastásico del PDA. Además, mediante técnicas glico-proteómicas, pudimos identificar la glicoproteína MFAP4 portadora de SLeX como potencial biomarcador de PDA, ya que esta glicoforma solo fue detectada en la cohorte de muestras de tejido de PDA
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Oltean, Gabriel. "From Current Collectors to Electrodes : Aluminium Rod Structures for Three-dimensional Li-ion Micro-battery Applications." Doctoral thesis, Uppsala universitet, Institutionen för kemi - Ångström, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-215482.
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The potential use of 3D aluminium nanorod structures as current collectors and negative electrodes for 3D Li-ion micro-batteries was studied based on the use of relatively simple and cost-effective electrochemical and sol-gel deposition techniques. Aluminium rod structures were synthesised by galvanostatic electrodeposition using commercial porous membranes as templates. It was shown that the use of a short (i.e., 50 ms long) potential pulse (i.e., -0.9 V vs. Al3+/Al) applied prior to a pulsed current electrochemical deposition gave rise to homogeneous deposits with more even rod heights. Electrophoretic and sol-gel deposition of TiO2 on the same substrates were also studied. The use of the sol-gel technique successfully resulted in a thin coating of amorphous TiO2 on the Al nanorod current collector, but with relatively small discharge capacities due to the amorphous character of the deposits. Electrophoretic deposition was, however, successful only on 2D substrates. Anodisation of titanium was used to prepare 3D TiO2 nanotube electrodes, with a nanotube length of 9 um and wall thickness of 50 nm. The electrodes displayed high and stable discharge capacities of 460 µAh/cm2 at a 0.1 C rate upon prolonged cycling with good rate capability. The 3D aluminium nanorod structures were tested as negative electrodes for Li-ion cells and the observed capacity fading was assigned to trapping of LiAl alloy inside the aluminium electrode caused by the diffusion of lithium into the electrode, rather than to pulverisation of the aluminium rods. The capacity fading effect could, however, be eliminated by decreasing the oxidation potential limit from 3.0 to 1.0 V vs. Li+/Li. A model for the alloying and dealloying of lithium with aluminium was also proposed. Finally, a proof-of-concept for a full 3D Li-ion micro-battery with electrodes of different geometries was demonstrated. The cell comprised a positive electrode, based on LiFePO4 deposited on a carbon foam current collector, with an area gain factor an order of magnitude larger than that for the Al nanorod negative electrode. This concept facilitates the balancing of 3D Li-ion cells as the positive electrode materials generally have significant lower specific energy densities than the negative electrodes.
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Kultima, Kim. "Transcriptomics and Proteomics Applied to Developmental Toxicology." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7921.
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Vidotto, Alessandra. "Marcadores protéicos do carcinoma epidermóide de cabeça e pescoço com fenótipo invasivo." Faculdade de Medicina de São José do Rio Preto, 2009. http://bdtd.famerp.br/handle/tede/125.
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Previous issue date: 2009-08-27The regional lymph nodes play a pivotal role in diagnosis, staging and management of head and neck squamous cell carcinomas (HNSCC). Despite their importance, detailed understanding of the probable mechanisms of lymphatic metastases has not been completely achieved. Subjects and Methods: We analyzed metastatic and normal lymph node tissues, as well as saliva and serum from sixth-two patients with HNSCC, and twenty-nine controls using two-dimensional electrophoresis, MALDI-Q-TOF and western blot. Results: Several proteins were found to be significantly increased in metastatic nodes, such as stratifin, glutathione S-transferase pi, apoliproteín A-I, alpha-1-microglobulin, disulfide isomerase, galectin, citokeratins, immunoglobulins, transtirretin, calciun-binding protein (família S100) and fat-binding protein (FABP). Among the down-regulated proteins in metastatic lymph nodes are calreticulin, tropomiosin 3, triosephosphate isomerase, piruvate quinase, anidrase carbonic, gamma actin, peroxiredoxin 2, profilin 1, gliceraldeyde 3- fosfato desidrogenase and heat shock proteins. These proteins are involved in epidermis development, cell proliferation, migration and adhesion, apoptosis, defense and inflammatory response and xenobiotic metabolism. Our data on the expression of heat shock proteins and enzymes of the glycolytic pathway suggest an effect of the lymph node environment in controlling tumor progression or in metabolic reprogramming of the metastatic cell. In saliva, 13 proteins showed an altered pattern of expression in samples patient, including over-expression of keratins, immunoglobulins, alphaamylase, PLUNC and zinc-alpha-2-glycoprotein and down-regulation of myosin. In serum samples, six proteins were over-expressed (serum albumin, alpha-1- microglobulin/bikunin precursor, apolipoprotein A-I, haptoglobin, serotransferrin, transthyretin) and two were under-expressed (hemoglobin subunit alpha, hemoglobin subunit beta) compared to the control group. Conclusion: New potential markers, such as profilin-1 and E-FABP, were identified and may be proved useful for defining the invasive phenotype of head and neck carcinomas.
O comprometimento de linfonodos regionais por células neoplásicas é atualmente o indicador mais utilizado para prognóstico em pacientes com carcinoma epidermóide de cabeça e pescoço (CECP). Apesar disso, a compreensão detalhada dos mecanismos envolvidos na formação de metástases linfáticas ainda não foi completamente atingida. Casuística e Método: Foi avaliado o perfil protéico de linfonodos metastáticos e não metastáticos, bem como de amostras de saliva e soro de 62 pacientes em diferentes estágios da doença e de 29 controles, utilizando eletroforese bidimensional, espectrometria de massas por MALDI-Q-TOF e experimentos de validação por Western blot. Resultados: Os resultados mostraram várias proteínas com expressão elevada em linfonodos metastáticos em relação aos não metastáticos, como stratifina, glutathiona S-transferase pi, apoliproteína A-I, alfa-1-microglobulina, dissulfeto isomerase, galectinas, citoqueratinas, imunoglobulinas, transtirretina e proteínas de ligação ao cálcio (família S100) e a ácidos graxos (FABP). De forma inversa, as proteínas calrreticulina, tropomiosina 3, triofosfato isomerase, piruvato quinase, anidrase carbônica, gama actina, peroxirredoxina 2, profilina 1, gliceraldeído 3-fosfato desidrogenase e proteínas de choque térmico mostraram níveis reduzidos em linfonodos metastáticos. Essas proteínas estão envolvidas em processos de desenvolvimento epidérmico, proliferação, migração e adesão celular, apoptose, resposta inflamatória e metabolismo de xenobióticos. Os dados relacionados à expressão de proteínas de choque térmico e enzimas da via glicolítica sugerem um efeito do ambiente dos linfonodos e no controle da progressão do tumor ou na reprogramação das células metastáticas. Em saliva, 13 proteínas exibiram um padrão alterado nas amostras de pacientes com câncer, incluindo expressão elevada de queratinas, imunoglobulinas, alfa-amilase, PLUNC e zinc-alfa-2-glicoproteína e expressão reduzida de miosina. Em amostras de soro, seis proteínas apresentaram expressão aumentada (albumina, alfa-1-microglobulina/bikunina precursor, apolipoproteína A-I, haptoglobina, serotransferrina e transtirretina) e duas estavam com expressão diminuída (hemoglobina alfa e hemoglobina beta), quando comparadas com o grupo controle. Conclusão: Os resultados obtidos revelaram novos marcadores potenciais, como profilina 1 e E-FABP, PLUNC e transtirretin que podem ser úteis na definição do fenótipo invasivo e no rastreamento e diagnóstico desse grupo de neoplasias.
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Bondy, Christina M. "Characterization and improvement of two-dimensional gel electrophoresis." 2005. http://catalog.hathitrust.org/api/volumes/oclc/64445818.html.
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