Evaluation of Lower Molecular Mass (10–30KDa) T. Solium cysticerci antigen by Western blotting for the diagnosis of neurocysticercosis in children

In order to elucidate the regulatory mechanisms of expression of the human endothelin-A receptor (hET-AR) gene, we characterized hET-AR transcripts using reverse transcriptase (RT)-PCR analysis in a variety of human tissues. RT-PCR of lung mRNA using a set of primers from exons 2 and 5 showed two lower-molecular-mass transcripts in addition to the expected fragment. When RT-PCR with primers from exons 4 and 8 was performed, no transcripts other than the expected one were detected. PCR cloning utilizing a set of primers from exons 2 and 8 which covered the entire coding sequence revealed that the cDNA clones corresponding to the two novel transcripts contained deletions of 199 bp and 327 bp respectively compared with the previously described hET-AR cDNA. Comparison of their sequences with that of the hET-AR gene showed that the deleted sequences correspond exactly to exon 4 and exons 3 and 4 respectively, indicating that these lower-molecular-mass ET-AR transcripts result from alternative RNA splicing (designated ET-AR∆4 and ET-AR∆3,4 respectively). Alternative splicing of exon 4 results in a transcript which would be translated into a C-terminal truncated protein containing the first, second and third transmembrane domains, while the splicing out of exons 3 and 4 would produce a protein with five membrane-spanning domains but lacking the third and fourth domains present in the ET-AR protein. An RNase protection assay revealed that ET-AR∆4 and ET-AR∆3,4, as well as ET-AR, transcripts were observed in various human tissues, including the lung, aorta, atrium, kidney and placenta, which are known to express ET-AR abundantly. Thus we have isolated the cDNAs of novel transcripts of hET-AR which are generated by alternative RNA splicing, and these results suggest that this alternative RNA splicing might contribute to the regulation of ET-AR gene expression.

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Microfluidic Western Blotting of Low-Molecular-Mass Proteins

Analytical Chemistry ◽

10.1021/ac5024588 ◽

2014 ◽

Vol 86 (21) ◽

pp. 10625-10632 ◽

Cited By ~ 14

Author(s):

Rachel E. Gerver ◽

Amy E. Herr

Keyword(s):

Molecular Mass ◽

Western Blotting

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Effect of diosgenin on biliary cholesterol transport in the rat

Biochemical Journal ◽

10.1042/bj2910793 ◽

1993 ◽

Vol 291 (3) ◽

pp. 793-798 ◽

Cited By ~ 27

Author(s):

A Thewles ◽

R A Parslow ◽

R Coleman

Keyword(s):

Bile Salt ◽

Molecular Mass ◽

Bile Salts ◽

Mass Fraction ◽

Gel Permeation Chromatography ◽

High Molecular Mass ◽

Cholesterol Transport ◽

Biliary Cholesterol ◽

Lower Molecular Mass ◽

Salt Depletion

Biliary cholesterol output in rats was stimulated over 3-fold by feeding diosgenin for 5 days, whereas biliary outputs of phospholipid and bile salts were not changed by diosgenin feeding. Isolating and perfusing the liver without bile salts resulted in a rapid and substantial decrease in biliary bile salt output; bile salt depletion abolished the diosgenin-induced increment in biliary cholesterol output, showing that the diosgenin-elevated biliary cholesterol output was bile-salt-dependent. Diosgenin treatment also produced a significant decrease in biliary alkaline phosphodiesterase I. Fresh bile obtained from control and diosgenin-fed rats was subjected to gel-permeation chromatography in order to separate different-sized biliary cholesterol carriers. Two major peaks of cholesterol were eluted, with cholesterol also being eluted between the peaks. The cholesterol peak eluted at the lower molecular mass (20-30 kDa) was observed in all bile samples. The higher-molecular-mass peak, which was eluted at the void volume, was not observed in all biles; control biles contained very little high-molecular-mass form of cholesterol, whereas biles from the diosgenin group contained up to 47% of cholesterol in the high-molecular-mass fraction. Diosgenin treatment produced a range of elevated biliary cholesterol values which positively correlated with the proportion of cholesterol contained in the high-molecular-mass fraction (r = 0.98). The results show that diosgenin induced a marked bile-salt-dependent increase in biliary cholesterol output and a shift in biliary cholesterol transport to higher-molecular-mass structures.

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Continuous s.c. infusion rather than twice-daily injections of IGF-I more effectively increases serum IGF binding protein-3 in female monkeys

Acta Endocrinologica ◽

10.1530/eje.0.1410303 ◽

1999 ◽

pp. 303-312 ◽

Cited By ~ 5

Author(s):

ME Wilson ◽

SL Lackey

Keyword(s):

Molecular Mass ◽

Ternary Complex ◽

Binding Protein ◽

Serum Insulin ◽

Size Exclusion ◽

Constant Infusion ◽

Serum Concentrations ◽

Lower Molecular Mass ◽

Igf I ◽

Igfbp 3

OBJECTIVE: In order to better understand how the IGF-I axis is affected by exogenous IGF-I, this study compared the effects of a constant s.c. infusion of IGF-I with that of twice-daily injections of IGF-I in young adult female rhesus monkeys. Clinical studies suggest that circulating concentrations of insulin-like growth factor binding protein-3 (IGFBP-3) are decreased or unaffected by IGF-I administration, whereas acute increases in IGF-I may increase serum IGFBP-1. However, studies in monkeys indicate that acute or continuous infusion of IGF-I effectively increases serum IGFBP-3. DESIGN AND METHODS: Female monkeys were studied for 5 days with no IGF-I supplementation (baseline) and for 5 days of IGF-I treatment by either constant infusion (120 microg/kg per day s.c., n = 5) or twice-daily injections of IGF-I (60 microg/kg per injection s.c., n = 5). Serum samples were collected daily at 0800 h and at 0800, 0900, 1100, 1500, and 2000 h on days 1 and 4 for each condition. Samples were assayed for IGF-I, IGFBPs-1 and -3, insulin, and glucose. RESULTS: Serum IGF-I was consistently increased above baseline within 24 h of the initiation of constant infusion, but was delayed until the second day of treatment in the injection group. Serum IGFBP-3 followed the pattern of IGF-I, with concentrations increased by day 1 during constant infusion and by day 2 during intermittent injections. Although both treatments effectively increased serum IGFBP-3, the increase was greater during constant infusion (31% above baseline) compared with injection (17%). Immunoblotting revealed that the constant infusion of IGF-I resulted in quantitatively more lower-molecular-mass fragments of IGFBP-3 than were observed during baseline or intermittent injections. Size-exclusion chromatography and ultrafiltration indicated that most IGFBP-3 was found in the ternary complex, with a greater percentage found in the ternary complex during baseline (90%) than during constant infusion (86%) or intermittent injections of IGF-I (87%). In contrast, serum concentrations of IGFBP-1 were increased on day 1 of both treatments, but declined towards baseline values as treatment progressed. Serum concentrations of insulin and glucose were unaffected by either mode of IGF-I treatment. Serum concentrations of IGF-I and IGFBP-3 were increased within 3h of the injection, before declining towards the pre-injection level. In contrast, the daily pattern of serum hormone concentrations was similar between the baseline condition and during constant infusion of IGF-I. Although higher during the treatment phase, serum IGF-I and IGFBP-3 concentrations decreased significantly from 0800 h until the afternoon meal, reaching a nadir in the evening before increasing again the next morning. Serum insulin decreased also after the morning meal and increased significantly immediately after the afternoon meal. Although serum IGFBP-1 also decreased initially after the morning meal, concentrations reached a peak before the afternoon meal as serum insulin reached its nadir. CONCLUSION: The results of the present analysis indicate that the constant infusion of IGF-I more effectively sustains serum concentrations of IGF-I and IGFBP-3 than do twice-daily injections. Although the percentage of IGF-I and IGFBP-3 in the ternary complex was similar during both treatments, the constant infusion regimen produced lower-molecular-mass fragments of IGFBP-3. In addition, serum IGF-I and IGFBP-3 appeared to be regulated diurnally, even during IGF-I infusion, whereas IGFBP-1 and insulin were affected by the timing of food intake. Taken together, these data suggest that, in the monkey, IGFBP-3 is regulated by factors in addition to GH, and that IGF-I can affect its own bioavailability by increasing circulating concentrations of IGFBP-3.

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Isotopic tracer studies of the further reactions of pentenes in the combusion of pentane

Proceedings of the Royal Society of London Series A - Mathematical and Physical Sciences ◽

10.1098/rspa.1978.0203 ◽

1978 ◽

Vol 364 (1718) ◽

pp. 309-329 ◽

Cited By ~ 8

Keyword(s):

Molecular Mass ◽

Peroxy Radical ◽

Combustion Products ◽

Secondary Products ◽

Tracer Studies ◽

Isotopic Tracer ◽

Cool Flames ◽

Hydroperoxy Radical ◽

Lower Molecular Mass ◽

Before And After

Detailed studies have been made of the mechanisms by which products other than the conjugate alkenes are formed when pentane undergoes combustion in the presence of small quantities of isotopically labelled pent-1-ene and pent-2-ene. Seven pentenes, specifically labelled with 14 C in different skeletal positions, have been synthesized and the fate of the labelled carbon atoms during combustion has been determined. Special attention has been paid to the formation and destruction of the pentadienes, acrolein and ethylene, as products derived from pent-1-ene. In particular, measurements have been made of the instantaneous fraction of the original pentene converted into each of these compounds and of the percentages have been shown to be derived exclusively from the pentenes, whilst acrolein and ethylene, which are complementary products resulting simultaneously from the same pentene molecule, are formed principally from the alkenes. It has also been possible to determine the distribution of the points of abstractive attack in the pentenes and the reactivity rations of pent-1-ene to secondary products. There are sharply defined changes in the relative rates for the pentadienes and pent-1-ene before and after the passage of cool flames; the results suggest that penta-1, 2-diene is oxidized to acrolein and ethylene. A significant amount of 2, 4-dimethyloxetan is formed from both pent-1-ene and pent-2-ene; this compound appears to be produced by a somewhat unusual route, involving the isomerization of one hydroperoxy radical to another through a peroxy radical. The majority of these compounds result from reactions involving free radial addition to the alkene molecule. However, abstractive modes of attack, although leading almost exclusively to the pentadienes, acrolein and ethylene, are quantitatively very important in terms of the total amount of attack on the pentenes. The principal products formed from the pentenes are only minor constituents of the combustion products of pentane. This shows that the coujugate alkenes do not play nearly as important a part in the combustion of pentane as they do in the case of alkanes of lower molecular mass.

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Thermal and Thermorheologic Characterization of Different Polyolefin Waste Fractions

Materials Science Forum ◽

10.4028/www.scientific.net/msf.907.74 ◽

2017 ◽

Vol 907 ◽

pp. 74-79

Author(s):

Adela Lazar ◽

Catalin Croitoru ◽

Mircea Horia Tierean ◽

Liana Sanda Baltes

Keyword(s):

Molecular Mass ◽

High Density Polyethylene ◽

Melt Flow Index ◽

Household Waste ◽

Flow Index ◽

Melt Flow ◽

Polymer Waste ◽

Lower Molecular Mass ◽

Low Shear

In this study, melt flow index values from several household waste fractions containing mainly polypropylene and high-density polyethylene, were measured at 190 °C for polyethylene and 230 °C for polypropylene-rich fractions. High values of MFI (low shear viscosities) have been reported probably due to the lower molecular mass of the polymer waste and/or the presence of surfactant compounds on the surface of the polymer flakes. Also, by extruding the same batch in different cycles at the same temperature values, the number of processing cycles on which the polymer could be recycled has been determined.

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Differential localization of tropomyosin isoforms in cultured nonmuscle cells.

The Journal of Cell Biology ◽

10.1083/jcb.107.2.563 ◽

1988 ◽

Vol 107 (2) ◽

pp. 563-572 ◽

Cited By ~ 95

Author(s):

J J Lin ◽

T E Hegmann ◽

J L Lin

Keyword(s):

Molecular Mass ◽

Chicken Embryo Fibroblast ◽

Polyclonal Antiserum ◽

Western Blots ◽

Tropomyosin Isoforms ◽

Lower Molecular Mass ◽

Nonmuscle Cells ◽

Double Label ◽

Sequential Absorption ◽

The Stability

We have previously shown that chicken embryo fibroblast (CEF) cells and human bladder carcinoma (EJ) cells contain multiple isoforms of tropomyosin, identified as a, b, 1, 2, and 3 in CEF cells and 1, 2, 3, 4, and 5 in human EJ cells by one-dimensional SDS-PAGE (Lin, J. J.-C., D. M. Helfman, S. H. Hughes, and C.-S. Chou. 1985. J. Cell Biol. 100: 692-703; and Lin, J. J.-C., S. Yamashiro-Matsumura, and F. Matsumura. 1984. Cancer Cells 1:57-65). Both isoform 3 (TM-3) of CEF and isoforms 4,5 (TM-4,-5) of human EJ cells are the minor isoforms found respectively in normal chicken and human cells. They have a lower apparent molecular mass and show a weaker affinity to actin filaments when compared to the higher molecular mass isoforms. Using individual tropomyosin isoforms immobilized on nitrocellulose papers and sequential absorption of polyclonal antiserum on these papers, we have prepared antibodies specific to CEF TM-3 and to CEF TM-1,-2. In addition, two of our antitropomyosin mAbs, CG beta 6 and CG3, have now been demonstrated by Western blots, immunoprecipitation, and two-dimensional gel analysis to have specificities to human EJ TM-3 and TM-5, respectively. By using these isoform-specific reagents, we are able to compare the intracellular localizations of the lower and higher molecular mass isoforms in both CEF and human EJ cells. We have found that both lower and higher molecular mass isoforms of tropomyosin are localized along stress fibers of cells, as one would expect. However, the lower molecular mass isoforms are also distributed in regions near ruffling membranes. Further evidence for this different localization of different tropomyosin isoforms comes from double-label immunofluorescence microscopy on the same CEF cells with affinity-purified antibody against TM-3, and monoclonal CG beta 6 antibody against TM-a, -b, -1, and -2 of CEF tropomyosin. The presence of the lower molecular mass isoform of tropomyosin in ruffling membranes may indicate a novel way for the nonmuscle cell to control the stability and organization of microfilaments, and to regulate the cell motility.

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PRODUCTION OF β-GLUCAN CONCENTRATE BY OAT GERMINATION

chemistry of plant raw material ◽

10.14258/jcprm.2019024251 ◽

2019 ◽

pp. 231-237 ◽

Cited By ~ 1

Author(s):

Venera Maratovna Gematdinova ◽

Al'bert Vladimirovich Kanarskiy ◽

Zosya Albertovna Kanarskaya ◽

Igor Vadimovich Kruchina-Bogdanov

Keyword(s):

Molecular Mass ◽

Functional Food ◽

Maximal Activity ◽

Industrial Processes ◽

Amylolytic Enzymes ◽

Naked Oat ◽

Lower Molecular Mass ◽

Polysaccharide Composition

Today industrial processes manufacturing functional food with elevated β-glucan fraction are vulnerable due to heavy losses in energy and valuable human nutrients, and that calls for new technological solutions. Oat malting is discussed as an approach to obtain β-glucan concentrates rich with protein, vitamins, enzymes by reducing starch level. Hulless (or naked) oat malting duration was evaluated to control polysaccharide composition in grains. Amylolytic enzymes complex was demonstrated to degrade starch in 15 days from 59.5% to 31%, with β-glucan level rising correspondingly from 6.1% to 16.0%. It was noted that up to 6% of β-glucan can be synthesized in sprouts. Simultaneously, sprouting was accompanied by activation of endo-β-1,3-glucanase in aleuronic layer, and the maximal activity was observed on the 6th to 7th days of malting. This enzyme converts β-glucans to oligomers with lower molecular mass — β-glucan oligosaccharides, with concentration of the latter reaching 6% on the 15th day of malting.

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IGF binding protein 5 and IGF-I receptor regulation in hypophysectomized rat kidneys

AJP Renal Physiology ◽

10.1152/ajprenal.1994.266.1.f147 ◽

1994 ◽

Vol 266 (1) ◽

pp. F147-F154 ◽

Cited By ~ 1

Author(s):

M. K. Hise ◽

N. M. Mantzouris ◽

J. S. Lahn ◽

M. S. Sheikh ◽

Z. M. Shao ◽

...

Keyword(s):

Molecular Mass ◽

Binding Protein ◽

Affinity Labeling ◽

Similar Data ◽

Cortical Tissue ◽

Factor I ◽

Lower Molecular Mass ◽

Igf I ◽

Igf Binding ◽

Igf Binding Protein

This study examines growth-regulating adaptations made by the proximal nephron in response to hypophysectomy (HYPX). Fourteen days after HYPX, circulating insulin-like growth factor I (IGF-I) levels were diminished, averaging 97 +/- 7 compared with 650 +/- 69 ng/ml in controls (n = 5, P < 0.001). Similar data were observed at day 7. Binding of 125I-IGF-I to isolated glomerular membranes and proximal tubule basolateral membranes (BLM) was increased in HYPX rats. Affinity labeling of membranes with 125I-IGF-I followed by electrophoresis on 6% polyacrylamide gels demonstrated two bands, one of approximately 140 kDa and another of > 200 kDa. The lower-molecular-mass protein, which has been identified as the alpha-subunit of the IGF-I receptor, and the higher-molecular-mass species were both upregulated by HYPX. Ligand blotting with IGF-I demonstrated a 31-kDa protein in both membranes, identified by immunostaining as IGF binding protein 5 (IGFBP-5), not IGFBP-1 or IGFBP-2. Affinity labeling documented an upregulation of this protein in both membranes after HYPX. Ligand blotting demonstrated a 31-kDa protein in HYPX cortical but not normal cortical or medullary cytosol that was immunostained with IGFBP-5 antibodies. RNA prepared from normal kidney cortical tissue demonstrated a 6.0-kb IGFBP-5 transcript, which was increased at day 14 after HYPX. Adaptations in the kidney after HYPX include an upregulation of the IGF-I receptor as well as IGFBP-5.(ABSTRACT TRUNCATED AT 250 WORDS)

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The purification of tissue inhibitor of metalloproteinases-2 from its 72 kDa progelatinase complex. Demonstration of the biochemical similarities of tissue inhibitor of metalloproteinases-2 and tissue inhibitor of metalloproteinases-1

Biochemical Journal ◽

10.1042/bj2780179 ◽

1991 ◽

Vol 278 (1) ◽

pp. 179-187 ◽

Cited By ~ 110

Author(s):

R V Ward ◽

R M Hembry ◽

J J Reynolds ◽

G Murphy

Keyword(s):

Molecular Mass ◽

Polyclonal Antiserum ◽

High Molecular Mass ◽

U937 Cells ◽

Tissue Inhibitor Of Metalloproteinases ◽

Gingival Fibroblasts ◽

Tissue Inhibitor ◽

Lower Molecular Mass ◽

Inhibitory Activities

Human gingival fibroblasts in culture were shown to secrete a 72 kDa progelatinase, of which a proportion in the medium was found to be complexed with tissue inhibitor of metalloproteinases-2 (TIMP-2). A purification procedure was devised to purify free enzyme and inhibitor. We also describe the purification of both 95 kDa progelatinase bound to TIMP-1 and free 95 kDa progelatinase from the medium of U937 cells. A polyclonal antiserum to TIMP-2 was prepared and it was shown that TIMP-1 and TIMP-2 are antigenically distinct. The ability to form stable complexes and the relative inhibitory activities of TIMP-1 and TIMP-2 towards 95 kDa and 72 kDa gelatinases, collagenase, stromelysins 1 and 2 and punctuated metalloproteinase were determined; only minor differences were found. Complex-formation between TIMP-2 and 72 kDa progelatinase was demonstrated not to reduce the metalloproteinase-inhibitory activity of TIMP-2, a finding that led to the characterization of high-molecular-mass TIMP activity. Competition experiments between progelatinases and active gelatinases for TIMPs indicated that the affinity of TIMPs for progelatinases is weaker than that for active gelatinases. In a study of the effects of TIMP-1 and TIMP-2 on progelatinase self-cleavage we found that both TIMP-1 and TIMP-2 inhibit the conversion of 95 kDa and 72 kDa progelatinases and prostromelysin into lower-molecular-mass forms. TIMP capable of complexing with progelatinase was shown to be no more efficient an inhibitor of gelatinase self-cleavage than TIMP not able to complex with progelatinase.

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