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Molecular Weight Proteins (molecular + weight_protein)
Kinds of Molecular Weight Proteins Selected AbstractsAn evolutionarily conserved gene required for proper microtubule architecture in Caenorhabditis elegansGENES TO CELLS, Issue 2 2004Satoshi Ogawa Microtubules are involved in many cellular events during the cell cycle and also in a variety of early embryonic developmental processes. Their architecture and properties change dramatically during the cell cycle and are properly regulated. However, these regulatory mechanisms have not been fully elucidated. C05D11.3 gene of Caenorhabditis elegans encodes a low molecular weight protein that is evolutionarily conserved from yeasts to mammals. A mouse homolog of the C05D11.3 product, APACD (ATP binding protein associated with cell differentiation), contains a thioredoxin-like domain and P-loop, and is present in both the nucleus and the cytoplasm, showing often localization to centrosomes and midbody. In C. elegans, C05D11.3 is expressed throughout development with higher levels of expression in most cells of the nervous system and in vulva. C05D11.3 RNAi-treated embryos show apparent defects in pronuclear migration or nuclear-centrosome rotation, and exhibit little astral microtubules and defective small spindles. These results indicate that C05D11.3, an evolutionarily conserved gene, is essential for proper microtubule organization and function in C. elegans. This gene family may be a conserved regulator of microtubule dynamics and function. [source] Shift of C3 deposition from localization in the glomerulus into the tubulo-interstitial compartment in the absence of secreted IgM in immune complex glomerulonephritisCLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 1 2008C. Vaculik Summary The role of secretory IgM in protecting kidney tissue from immune complex glomerulonephritis induced by 4 mg horse spleen apoferritin and 0·05 mg lipopolysaccharide has been investigated in mutant mice in which B cells do not secrete IgM, but are capable of expressing surface IgM and IgD and secreting other Ig isotypes. Glomerular size, number of glomeruli per cross-section, glomerular cellularity and urine content of protein and creatinine was comparable in treated secreted IgM (sIgM)-deficient and wild-type mice. Assessment of urinary proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed a 30 kDa low molecular weight protein in treated sIgM-deficient animals only, reflecting dysfunction of proximal tubules. A shift of bound C3 from glomeruli to the tubulo-interstitial compartment in sIgM-deficient mice also suggests tubulo-interstitial damage. In contrast, local C3 synthesis within the kidney tissue did not differ between the two treated groups. Apoptosis physiologically present to maintain kidney cell homeostasis was increased slightly in treated wild-type mice. These results indicate that secretory IgM can protect the tubulo-interstitial compartment from immune complex-induced damage without having an effect on the glomerulus. [source] Improved 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] Identification of a distinct family of genes encoding atypical odorant-binding proteins in the malaria vector mosquito, Anopheles gambiaeINSECT MOLECULAR BIOLOGY, Issue 6 2003P. X. Xu Abstract We performed a genome-wide analysis for candidate odorant-binding protein (OBP) genes in the malaria vector Anopheles gambiae (Ag). We identified fifty-seven putative genes including sixteen genes predicted to encode distinct, higher molecular weight proteins that lack orthologues in Drosophila. Expression analysis indicates that several of these atypical AgOBPs are transcribed in chemosensory organs in adult and immature stages. Phylogenetic analysis of the Anopheles and Drosophila OBP families reveals these proteins fall into several clusters based on sequence similarity and suggests the atypical AgOBP genes arose in the mosquito lineage after the divergence of mosquitoes and flies. The identification of these AgOBP genes is the first step towards determining their biological roles in this economically and medically important insect. [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] Statins and progressive renal diseaseMEDICINAL RESEARCH REVIEWS, Issue 1 2002Michele Buemi Abstract Thanks to the administration of hypocholesterolemic drugs, important advances have been made in the treatment of patients with progressive renal disease. In vitro and in vivo findings demonstrate that statins, the inhibitors of HMG-CoA reductase, can provide protection against kidney diseases characterized by inflammation and/or enhanced proliferation of epithelial cells occurring in rapidly progressive glomerulonephritis, or by increased proliferation of mesangial cells occurring in IgA nephropathy. Many of the beneficial effects obtained occur independent of reduced cholesterol levels because statins can directly inhibit the proliferation of different cell types (e.g., mesangial, renal tubular, and vascular smooth muscle cells), and can also modulate the inflammatory response, thus inhibiting macrophage recruitment and activation, as well as fibrosis. The mechanisms underlying the action of statins are not yet well understood, although recent data in the literature indicate that they can directly affect the proliferation/apoptosis balance, the down-regulation of inflammatory chemokines, and the cytogenic messages mediated by the GTPases Ras superfamily. Therefore, as well as reducing serum lipids, statins and other lipid-lowering agents may directly influence intracellular signaling pathways involved in the prenylation of low molecular weight proteins that play a crucial role in cell signal transduction and cell activation. Statins appear to have important potential in the treatment of progressive renal disease, although further studies are required to confirm this in humans. © 2001 John Wiley & Sons, Inc. Med Res Rev, 22, No. 1, 76,84, 2002 [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] Altered actin polymerization of Plutella xylostella (L.) in response to ovarian calyx components of an endoparasitoid Cotesia plutellae (Kurdjumov)PHYSIOLOGICAL ENTOMOLOGY, Issue 2 2009MADANAGOPAL NALINI AbstractCotesia plutellae (Kurdjumov) (Hymenoptera: Braconidae), a solitary braconid endoparasitoid wasp, parasitizes the diamondback moth Plutella xylostella (L.) (Lepidoptera: Yponomeutidae) by suppressing the host defense response, thereby resulting in successful parasitization. During parasitization, ovarian calyx fluid is also delivered into the haemocoel of the host along with the wasp egg. The effect of calyx fluid constituents on haemocyte-spreading behaviour of P. xylostella is analysed by measuring F-actin development in the haemocytes. For this purpose, the calyx fluid of C. plutellae is separated into ovarian protein and C. plutellae bracovirus (CpBV). The ovarian protein consists of a wide range of molecular weight proteins, which are apparently different from those of CpBV. When nonparasitized P. xylostella haemocytes are incubated with either ovarian protein or CpBV for 1 or 2 h, haemocytes lose their responsiveness to a cytokine, plasmatocyte-spreading peptide, in a dose-dependent manner for each calyx component and fail to exhibit haemocyte-spreading behaviour. Some CpBV genes are expressed within 1 h of parasitization. The inhibition of haemocyte-spreading could be explained by measuring F-actin contents, in which parasitization by C. plutellae inhibits F-actin development in the haemocytes of P. xylostella. Either ovarian protein or CpBV could inhibit F-actin development in the nonparasitized haemocytes. In addition, co-incubation of ovarian protein and CpBV results in significant additive inhibition of both haemocyte-spreading and F-actin development in the haemocytes in response to cytokine. These results suggest that both components of C. plutellae calyx fluid function in a synergistic manner, leading to immunosuppression during the early stage of parasitization. [source] The nutritional and metabolic indices in rats fed cholesterol-containing diets supplemented with durian at different stages of ripeningBIOFACTORS, Issue 2-3 2007Maria Leontowicz Abstract The aim of this investigation was to assess the nutritional and health properties of Mon Thong durian cultivar at different stages of ripening. The assessment was carried out in vitro and in vivo. The contents of dietary fibers, minerals and trace metals at different stages of ripening were comparable. Total polyphenols (mgGAE/100 g FW) and flavonoids (mg CE/100 gFW) in ripe durian (358.8 ± 31.4 and 95.4 ± 9.3) were significantly higher (p < 0.05) than in mature (216.1 ± 1 and 39.9 ± 3.8) and overripe (283.3 ± 26.2 and 53.5 ± 4.9). Antioxidant capacity (,MTE/100 g FW) in total polyphenol extracts of ripe durian measured by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)] (ABTS) assays (259.4 ± 23.6 and 2341.8 ± 93.2) were significantly higher (p < 0.05) than that of mature (151.6 ± 15.2 and 1394.6 ± 41.5) and overripe (201.7 ± 19.4 and 1812.2 ± 61.4) samples. The correlation coefficients between the bioactive compounds in different stages of ripening and their antioxidant capacities were high (R2 = 0.99). Then 35 male Wistar rats were divided into 5 dietary groups each of 7 and named Control, Chol, Chol/Mature, Chol/Ripe and Chol/Overripe. During 30 days of the experiment the rats of all 5 groups were fed basal diet (BD), which included wheat starch, casein, soybean oil, vitamin and mineral mixtures. The rats of the Control group were fed a BD only. To the BD of the Chol group was added 1% of cholesterol. The BD of the Chol/Mature, Chol/Ripe and Chol/Overripe groups was supplemented with 1% of cholesterol and 5% of the mature, ripe and overripe durian as freeze-dried powder, respectively. Diets containing ripe and to a lesser degree mature and overripe durian significantly hindered the rise in plasma lipids and also hindered a decrease in plasma antioxidant activity. The nitrogen retention in rats of the Chol/Ripe group was significantly higher (63.6%, P < 0.05) than in other diet groups and the level of the plasma glucose remained normal. A decrease in fibrinogen fraction with ripe durian included in rat's diets was shown by electrophoretic separation. These changes were detected mostly in the low molecular weight proteins of rat's serum. Histological examination of aorta showed only slight differences in the tissue. In conclusion, ripe durian contains higher quantity of bioactive compounds, has higher antioxidant capacity and nutritional value. It positively affects the plasma lipid profile, the plasma glucose and the antioxidant activity in rats fed cholesterol enriched diets. Therefore, the ripe durian supplemented diet could be beneficial for patient suffering from hypercholesterolemia and diabetes mellitus. [source] Backbone-Only Protein Solution Structures with a Combination of Classical and Paramagnetism-Based Constraints: A Method that Can Be Scaled to Large MoleculesCHEMPHYSCHEM, Issue 6 2004Renato Barbieri Dr. Abstract Herein, it is shown that a medium-resolution solution structure of a protein can be obtained with the sole assignment of the protein backbone and backbone-related constraints if a derivative with a firmly bound paramagnetic metal is available. The proof-of-concept is provided on calbindin D9k, a calcium binding protein in which one of the two calcium ions can be selectively substituted by a paramagnetic lanthanide ion. The constraints used are HN (and H,) nuclear Overhauser effects (NOEs), hydrogen bonds, dihedral angle constraints from chemical shifts, and the following paramagnetism-based constraints: 1) pseudocontact shifts, acquired by substituting one (or more) lanthanide(s) in the C-terminal calcium binding site; 2) NHN residual dipolar couplings due to self-orientation induced by the paramagnetic lanthanide(s); 3) cross-correlations between the Curie and internuclear dipole,dipole interactions; and 4) paramagnetism-induced relaxation rate enhancements. An upper distance limit for internuclear distances between any two backbone atoms was also given according to the molecular weight of the protein. For this purpose, the paramagnetism-based constraints were collectively implemented in the program CYANA for solution structure determinations, similarly to what was previously done for the program DYANA. The method is intrinsically suitable for large molecular weight proteins. [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] | ||||||||||||||||