Immunoextracted calcitonin in milk and plasma from totally thyroidectomized women. Evidence of monomeric calcitonin in plasma during pregnancy and lactation

1986 ◽  
Vol 113 (4) ◽  
pp. 529-535 ◽  
Author(s):  
Elisabet Bucht ◽  
Margareta Telenius-Berg ◽  
Göran Lundell ◽  
Hans-Erik Sjöberg
Keyword(s):  
Molecular Weight ◽  
Mammary Gland ◽  
High Molecular Weight ◽  
Gel Chromatography ◽  
C Cells ◽  
Local Production ◽  
Thyroid C Cells ◽  
Pregnancy And Lactation ◽  
Series Of Experiments ◽  
High Concentrations

Abstract. The level of immunoreactive calcitonin in the first produced breast milk was in totally thyroidectomized (TX) women 713 ± 307 pg-eq/ml (mean ± sd, N = 7) and in control women 772 ± 329 pg-eq/ml (N = 33), i.e. about 45 times higher than in plasma (see below). On gel chromatography of immunoextracted milk from TX women, immunoreactive calcitonin appeared as high molecular weight forms in the same pattern as in milk from healthy women when the same antiserum (1) was used for immunoextraction and radioimmunoassay (RIA). In another series of experiments, a new antiserum (2) raised in rabbits was used for measurement of immunoreactive calcitonin after immunoextraction with 1. Plasma levels of immunoreactive calcitonin in the TX women during pregnancy were 16 ± 6 pg-eq/ml (N = 6) and during lactation 14 ± 7 pg-eq/ml (N = 5). Immunoreactive calcitonin was undetectable (< 8 pg/ml) in plasma from those TX women in whom lactation had stopped (N = 5). Immunoextraction and gel chromatography of plasma collected during pregnancy and lactation from the TX women showed that the immunoreactive calcitonin present eluted in the region of monomeric calcitonin with both antiserum 1 and 2. In conclusion, high concentrations of high molecular weight forms of immunoreactive calcitonin have been demonstrated in milk from TX patients, most probably devoid of any calcitonin-producing thyroid C-cells. This points to a local production site in the mammary gland. Relatively high levels of immunoreactive calcitonin in plasma during pregnancy and lactation in TX women also indicate extrathyroidal production.

Studies on Soluble Fibrin Complexes as a Function of pH

1975 ◽  
Author(s):  
Y. Benabid ◽  
E. Concord ◽  
M. Suscillon
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Gel Chromatography ◽  
Agarose Gel ◽  
Monomer Mixture ◽  
Soluble Fibrin ◽  
Molecular Weights ◽  
Fibrin Monomer

Purified fibrinogen solutions, incubated with thrombin. CNBr. Sepharose, were subjected to agarose gel chromatography and eluted at different pH (6.5; 7.5; 8.5). Among high molecular weight derivatives formed by thrombin, the major component was a dimer. Gel chromatography at pH 8.5 showed a complexes peak distinct of that from fibrinogen, whereas at pH 6.5, only the fibrinogen peak appeared: fibrin monomer was eluted with fibrinogen as demonstrated by polyacrylamid gel electrophoresis 3.75% pH 8.9. SDS urea electrophoresis after reduction indicated that complexes peak contained two α-chains (α and α′). When fibrinogen was incubated with thrombin in the presence of FSF and calcium, several derivatives with higher and higher molecular weights were formed besides the dimer, and elution profiles of chromatography were identical at pH 6.5 and 8.5, thus indicating stable complexes formation. If fibrinogen-fibrin monomer mixture was subjected to FSF action at different pH, no complexes were formed at pH 6.5. These results confirm that at pH 6.5, any association was prevented.

scholarly journals Structure Analysis and Anti-Tumor and Anti-Angiogenic Activities of Sulfated Galactofucan Extracted from Sargassum thunbergii

Marine Drugs ◽  
2019 ◽  
Vol 17 (1) ◽  
pp. 52 ◽  
Author(s):  
Weihua Jin ◽  
Wanli Wu ◽  
Hong Tang ◽  
Bin Wei ◽  
Hong Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Molecular Weight ◽  
High Molecular Weight ◽  
Structure Analysis ◽  
Nmr Spectra ◽  
Low Molecular Weight ◽  
Gel Chromatography ◽  
Sargassum Thunbergii ◽  
Sulfation Pattern ◽  
Fucose Content

Sulfated galactofucan (ST-2) was obtained from Sargassum thunbergii. It was then desulfated to obtain ST-2-DS, and autohydrolyzed and precipitated by ethanol to obtain the supernatant (ST-2-S) and precipitate (ST-2-C). ST-2-C was further fractionated by gel chromatography into two fractions, ST-2-H (high molecular weight) and ST-2-L (low molecular weight). Mass spectrometry (MS) of ST-2-DS was performed to elucidate the backbone of ST-2. It was shown that ST-2-DS contained a backbone of alternating galactopyranose residues (Gal)n (n ≤ 3) and fucopyranose residues (Fuc)n. In addition, ST-2-S was also determined by MS to elucidate the branches of ST-2. It was suggested that sulfated fuco-oligomers might be the branches of ST-2. Compared to the NMR spectra of ST-2-H, the spectra of ST-2-L was more recognizable. It was shown that ST-2-L contain a backbone of (Gal)n and (Fuc)n, sulfated mainly at C4 of Fuc, and interspersed with galactose (the linkages were likely to be 1→2 and 1→6). Therefore, ST-2 might contain a backbone of (Gal)n (n ≤ 3) and (Fuc)n. The sulfation pattern was mainly at C4 of fucopyranose and partially at C4 of galactopyranose, and the branches were mainly sulfated fuco-oligomers. Finally, the anti-tumor and anti-angiogenic activities of ST-2 and its derivates were determined. It was shown that the low molecular-weight sulfated galactofucan, with higher fucose content, had better anti-angiogenic and anti-tumor activities.

"Colloidal" Phosphorus Errors in Gel Chromatography

1983 ◽  
Vol 40 (10) ◽  
pp. 1614-1621 ◽  
Author(s):  
B. Kent Burnison
Keyword(s):  
Molecular Weight ◽  
Acid Hydrolysis ◽  
High Molecular Weight ◽  
Gel Chromatography ◽  
Chemical Analyses ◽  
Molybdophosphoric Acid ◽  
Molybdenum Blue ◽  
Soluble Reactive Phosphate ◽  
Colloidal Phosphorus ◽  
Hydrolysis Of

Gel chromatography has been used for the separation of 32PO4 and a high molecular weight "colloidal" 32P-labeled fraction from 32PO4-labeled lakewater. When the labeled filtrate is treated with reagents required for the molybdenum blue method for orthophosphate analysis, only a small fraction of the "colloidal" peak is hydrolyzed to orthophosphate. As the reduced molybdophosphoric acid is strongly adsorbed to the dextran gel, quantitative elution of 32PO4 can be achieved with 0.05 mol∙L−1 NaOH and 0.3% NaCl. In hardwater lakes, care must be taken to eliminate the possibility of orthophosphate precipitation at higher pH. In these lakes, it is unlikely that the discrepancy between 32PO4 bioassays and chemical analyses can be solely attributed to acid hydrolysis of "colloidal" phosphorus. Microparticulate apatite also has the potential to release soluble reactive phosphate when the acidic molybdenum blue method is used.

scholarly journals Ein makroporöses Polyacrylamid für immunologische Diffusionstechniken / Macroporous Polyacrylamide, its Use for Immunological Diffusion Techniques

1972 ◽  
Vol 27 (6) ◽  
pp. 683-688 ◽  
Author(s):  
Helmut Determann ◽  
Axel Walch
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Monomer Concentration ◽  
Agarose Gels ◽  
Γ Globulins ◽  
Sample Application ◽  
Total Monomer Concentration ◽  
High Molecular Weight Proteins ◽  
High Concentrations

Gel layers of crosslinked polyacrylamide with different porosities were synthesized and their properties compared. At high concentrations of bifunctional monomers macroporous gels were formed, one of which (total monomer concentration 14%, methylene-bis-acrylamide 40%) permitted even high molecular weight proteins (e. g. γ-globulins) to diffuse more readily than in 2 per cent agar or agarose gels. Different immunological techniques were modified for application with this polymer. It offers advantages especially with respect to porosity, storability and sample application for quantitative diffusion techniques.

Factor VIII, Small Active Factor VIII Fragments, and von Willebrand Factor

1975 ◽  
Author(s):  
R. H. Wagner
Keyword(s):  
Molecular Weight ◽  
Factor Viii ◽  
High Molecular Weight ◽  
Small Molecule ◽  
Gel Chromatography ◽  
Carrier Protein ◽  
High Molecular Weight Fraction ◽  
Factor Viii Activity ◽  
Von Willebrand ◽  
Active Fragment

Factor VIII activity of human, canine, and bovine preparations is associated with a molecule of high molecular weight. These preparations dissociate almost completely in 0.25 M Ca2+, yielding a small active fragment with factor VIII activity, and a high molecular weight fraction (carrier protein) with almost no factor VIII activity. With Ca2+ absent, the small active fragment recombines with the carrier protein to form a large protein with factor VIII activity. Mixtures of normal human small active fragment with human hemophilic carrier protein give normal recombination. Comparable experiments with human von Willebrand and canine “carrier protein fractions” show no recombination. The data suggest a working model of a small active fragment(s) linked by noncovalent bonds to a carrier molecule, comprised of repeating subunits, deficient in von Willebrand’s disease.Thrombin increases the activity of factor VIII preparations or the small active fragment obtained by Ca2+ dissociation. When a factor VIII preparation is treated with thrombin and subjected to gel chromatography or ultracentrifugation, the factor VIII activity appears to be associated with a small molecule. Factor VIII activity of this small molecule is not enhanced by thrombin. These and other data will be discussed.

scholarly journals High-molecular-weight nuclear polyadenylate-containing ribonucleic acid isolated from the lactating guniea-pig mammary gland contains milk-protein messenger ribonucleic acid sequences

1979 ◽  
Vol 181 (2) ◽  
pp. 501-504 ◽  
Author(s):  
I C Bathurst ◽  
R K Craig ◽  
P N Campbell
Keyword(s):  
Molecular Weight ◽  
Mammary Gland ◽  
High Molecular Weight ◽  
Ribonucleic Acid ◽  
Milk Protein ◽  
Gradient Centrifugation ◽  
Nucleic Acid Hybridization ◽  
Nuclear Fraction ◽  
Messenger Ribonucleic Acid ◽  
Precursor Rna

1. Steady-state poly(A)-containing nuclear RNA was isolated from the lactating guinea-pig mammary gland and analysed by sucrose-gradient centrifugation under denaturing conditions. 2. Nucleic acid-hybridization studies demonstrated the presence of small amounts of high-molecular-weight RNA species containing milk-protein mRNA sequences sedimenting at 25S. These were not found in the post-nuclear supernatant, where milk-protein sequences sedimented only between 12S and 15S; these were also the predominant species in the nuclear fraction. 3. The results are consistent with the transcription of milk-protein genes as high-molecular-weight precursor RNA species, 3-4 times as large as the active mRNA species isolated from the post-nuclear fraction.

Distribution and Biological Availability of Reactive High Molecular Weight Phosphorus in Natural Waters in New Zealand

1980 ◽  
Vol 37 (4) ◽  
pp. 664-669 ◽  
Author(s):  
E. White ◽  
G. Payne
Keyword(s):  
Molecular Weight ◽  
New Zealand ◽  
High Molecular Weight ◽  
Natural Waters ◽  
Phosphorus Uptake ◽  
Algal Growth ◽  
Gel Chromatography ◽  
Growth Responses ◽  
Eutrophic Lakes ◽  
Reactive Phosphorus

Eutrophic lakes and many streams on the central volcanic plateau of the North Island, New Zealand, contain dissolved chemically reactive phosphorus which is not orthophosphate. Sephadex gel (G25-150) chromatograms reveal that one reactive component is of high molecular weight (MW > 5000) while a second component elutes in the same way as phosphate-phosphorus (PO4-P). The reactive high molecular weight phosphorus (RHMW-P) in streams generally forms only a small proportion of the dissolved reactive phosphorus (DRP). In some eutrophic lakes the proportion of RHMW-P in the DRP is high (70–80%). Each lake may have a characteristic distribution of the two forms of reactive phosphorus. In summer where DRP concentrations are low (<2 mg∙m−3) in lake surface waters, PO4-P seems to dominate over RHMW-P which is consistent with long 32PO4 turnover times which are found in many of the central volcanic plateau lakes. Algal growth responses to phosphorus uptake were similar for PO4-P and RHMW-P, but not all of the RHMW-P was taken up by Chlorella in bioassays. The proportion of the RHMW-P taken up by Chlorella was different for each lake examined, posing problems in relating DRP concentrations to algal growth responses in lakes.Key words: reactive phosphorus, phosphate-phosphorus, algae, Chlorella, growth, bioassay, freshwater analysis, molybdenum blue method, Sephadex gel chromatography

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