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High Molecular Weight Proteins (high + molecular_weight_protein)
Selected AbstractsImproved disc SDS-PAGE for extraction of low molecular weight proteins from serumELECTROPHORESIS, Issue 6 2010Tiechun Li Abstract The low molecular weight proteins can provide a lot of valuable information of biomarkers. To study these proteins, the high abundance and high molecular weight proteins must be removed prior to analysis. In this work, a simple and inexpensive disc SDS-PAGE to extract low molecular weight proteins from human serum and cutoff proteins larger than 30,kDa was developed. Some experimental conditions were examined. The experimental results obtained by plate SDS-PAGE and MALDI-TOF MS showed that the molecular weight of extracted proteins was about in the range from 0.3 to 28,kDa. Some experiments, including precipitation of proteins in organic solvents, SPE and cytochrome C test, were carried out and the experimental results demonstrated successful recovery of proteins/peptides with molecular weight from several hundreds of dalton to about 30,kDa. The experimental results obtained by plate SDS-PAGE indicated the repeatability was satisfactory. [source] A novel methodology for the analysis of membrane and cytosolic sub-proteomes of erythrocytes by 2-DEELECTROPHORESIS, Issue 23 2009Gloria Alvarez-Llamas Abstract With the aim of studying a wide cohort of erythrocyte samples in a clinical setting, we propose here a novel approach that allows the analysis of both human cytosolic and membrane sub-proteomes. Despite their simple structure, the high content of hemoglobin present in the red blood cells (RBCs) makes their proteome analysis enormously difficult. We investigate here different strategies for isolation of the membrane and cytosolic fractions from erythrocytes and their influence on proteome profiling by 2-DE, paying particular attention to hemoglobin removal. A simple, quick and satisfactory approach for hemoglobin depletion based on HemogloBindÔ reagent was satisfactorily applied to erythrocyte cells, allowing the analysis of the cytosolic sub-proteome by 2-DE without major interference. For membrane proteome, a novel combined strategy based on hypotonic lysis isolation and further purification on minicolumns is described here, allowing detection of high molecular weight proteins (i.e. spectrin, ankyrin) and well-resolved 2-DE patterns. An aliquot of the membrane fraction was also in solution digested and analyzed by nano-LC coupled to an LTQ-Orbitrap mass spectrometer. A total of 188 unique proteins were identified by this approach. This study sets the basis for future clinical studies where the erythrocyte cell may be implicated. [source] Enamel matrix derivative enhances tissue formation around scaffolds used for tissue engineering of ligamentsJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 2 2010Michael P. Messenger Abstract The following in vitro translational study investigated whether enamel matrix derivative (EMD), an approved biomimetic treatment for periodontal disease (Emdogain®) and hard-to-heal wounds (Xelma®), enhanced synovial cell colonization and protein synthesis around a scaffold used clinically for in situ tissue engineering of the torn anterior cruciate ligament (ACL). Synovial cells were enzymatically extracted from bovine synovium and dynamically seeded onto polyethylene terephthalate (PET) scaffolds. The cells were cultured in low-serum medium (0.5% FBS) for 4 weeks with either a single administration of EMD at the start of the 4 week period or multiple administrations of EMD at regular intervals throughout the 4 weeks. Samples were harvested and evaluated using the Hoechst DNA assay, BCA protein assay, cresolphthalein complexone calcium assay, SDS,PAGE, ELISA and electron microscopy. A significant increase in cell number (DNA) (p < 0.01), protein content (p < 0.01) and TGF,1 synthesis (p < 0.01) was observed with multiple administrations of EMD. Additionally, SDS,PAGE showed an increase in high molecular weight proteins, characteristic of the fibril-forming collagens. Electron microscopy supported these findings, showing that scaffolds treated with multiple administrations of EMD were heavily coated with cells and extracellular matrix (ECM) that enveloped the fibres. Multiple administrations of EMD to synovial cell-seeded scaffolds enhanced the formation of tissue in vitro. Additionally, it was shown that EMD enhanced TGF,1 synthesis of synovial cells, suggesting a potential mode of action for EMD's capacity to stimulate tissue regeneration. Copyright © 2009 John Wiley & Sons, Ltd. [source] Antigen selection for future anti- Trichuris vaccines: a comparison of cytokine and antibody responses to larval and adult antigen in a primary infectionPARASITE IMMUNOLOGY, Issue 9 2008H. DIXON SUMMARY Trichuriasis, caused by the whipworm Trichuris trichiura, is endemic in tropical and subtropical areas, affecting approximately 1 billion people. Child anthelminthic treatment programmes are being implemented but repeated treatments are costly, may prevent the development of acquired immunity and can lead to the development of drug resistant parasites. Thus, the development of a vaccine which would lead to the acquisition of immunity at an earlier age and reduce community faecal egg output would be beneficial. Development of subunit vaccines requires the identification of protective antigens and their formulation in a suitable adjuvant. Trichuris muris is an antigenically similar laboratory model for T. trichiura. Subcutaneous vaccination with adult excretory,secretory products (ES) protects susceptible mouse strains from T. muris. Larval stages may contain novel and more relevant antigens which when incorporated in a vaccine induce worm expulsion earlier in infection than the adult worm products. This study finds negligible difference in the cellular and humoral immune response to T. muris adult and third stage larva(e) (L3) ES during a primary T. muris infection, but identifies high molecular weight proteins in both adult and L3 ES as potential vaccine candidates. [source] The prognosis of occupational asthma due to detergent enzymes: clinical, immunological and employment outcomesCLINICAL & EXPERIMENTAL ALLERGY, Issue 4 2006A. Brant Summary Background Little is known about the prognosis of occupational asthma induced by high molecular weight proteins. Objective Our objective was to measure the clinical, immunological and employment outcomes of individuals with occupational asthma induced by detergent enzymes. Methods We undertook a workforce-based follow-up study in 35 (78%) of the 45 ex-employees from a single factory with occupational asthma. In each case the diagnosis was supported by evidence of specific sensitization and characteristic changes in peak flow or a positive response to specific bronchial provocation testing. Results This group had left the factory on average 37 months before study. On review 25 (71%) reported chest symptoms during the last month. Compared with when working at the factory, most (86%) reported that their symptoms had improved. Twenty continued to attend their general practitioner for respiratory symptoms and 19 still used asthma medications. Since leaving the factory 16 (46%) and four (11%) had found full-time or part-time employment, respectively; of these 16 found they were paid less than when they worked at the factory. The remaining 15 subjects had not had any paid employment. All but two had positive skin prick tests to one or more three detergent enzymes. The estimated half-life of serum-specific IgE antibodies was 20 months for protease, and 21 months for cellulase and amylase. Conclusions Population-based follow-up studies of the prognosis of occupational asthma are rare but probably avoid the bias in clinic-derived surveys. This study demonstrates that 3 years after the avoidance of exposure with detergent enzymes most patients continue to be troubled by, albeit improved, symptoms and experience difficulty in re-employment. [source] | ||||||||||