Identification of N-Monoacetylcystine in Uraemic Plasma

1981 ◽  
Vol 60 (3) ◽  
pp. 331-334 ◽  
Author(s):  
F. Gejyo ◽  
G. Ito ◽  
Y. Kinoshita
Keyword(s):  
Plasma Levels ◽  
Gel Filtration ◽  
Normal Subjects ◽  
Exchange Chromatography ◽  
Cysteic Acid ◽  
Homocysteic Acid ◽  
Normal Limits ◽  
Acidic Nature ◽  
Sulphur Amino Acids

1. An unidentified ninhydrin-positive substance of an acidic nature was detected in the plasma of uraemic patients. This substance was isolated from haemodialysate by ion-exchange chromatography and gel filtration, and identified as a sulphur-containing amino acid: N-monoacetylcystine. 2. The quantitative determination of sulphur amino acids in plasma revealed that the plasma levels of cysteic acid, homocysteic acid, taurine, cystine and cystathionine as well as N-monoacetylcystine in uraemic patients were markedly higher than in normal subjects (P < 0.001 for each). However, the plasma levels of methionine in uraemic patients were within normal limits.

scholarly journals Separation and partial characterization of guinea-pig caseins

1978 ◽  
Vol 173 (2) ◽  
pp. 633-641 ◽  
Author(s):  
R K Craig ◽  
D McIlreavy ◽  
R L Hall
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Guinea Pig ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Exchange Chromatography

1. Guinea-pig caseins A, B and C were purified free of each other by a combination of ion-exchange chromatography and gel filtration. 2. Determination of the amino acid composition showed all three caseins to contain a high proportion of proline and glutamic acid, but no cysteine. This apart, the amino acid composition of the three caseins was markedly different, though calculated divergence values suggest that some homology may exist between caseins A and B. Molecular-weight estimates based on amino acid composition were in good agreement with those based on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 3. N-Terminal analysis showed lysine, methionine and lysine to be the N-terminal residues of caseins A, B and C respectively. 4. Two-dimensional separation of tryptic digests revealed a distinctive pattern for each casein. 5. All caseins were shown to be phosphoproteins. The casein C preparation also contained significant amounts of sialic acid, neutral and amino sugars. 6. The results suggest that each casein represents a separate gene product, and that the low-molecular-weight proteins are not the result of a post-translational cleavage of the largest. All were distinctly different from the whey protein alpha-lactalbumin.

IMMUNOREACTIVE GROWTH HORMONE IN HUMAN URINE

1972 ◽  
Vol 71 (4) ◽  
pp. 665-676 ◽  
Author(s):  
Kristian F. Hanssen
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Linear Relationship ◽  
Human Urine ◽  
Gel Filtration ◽  
Human Growth ◽  
Normal Subjects ◽  
Normal Human ◽  
Radio Immunoassay

ABSTRACT By using a double antibody radio-immunoassay (pre-precipitation technique) for the determination of immunoreactive human growth hormone (IRHGH) in normal human urine concentrated by dialysis and lyophilization, a factor was revealed that displaces 125I-HGH from HGH antibodies. This displacement was neither due to salts nor to glucose; it is suggested that it is due to IRHGH in the urine. A linear relationship between dilution of urine and the measured IRHGH concentration was obtained. Recovery of exogenous HGH was between 70–105%. The recovery of IRHGH from different volumes of urine following dialysis and lyophilization was between 97–110%. Plasma IRHGH and urinary IRHGH was measured simultaneously after HGH injection in a normal subject. A correlation was shown between plasma IRHGH and urinary IRHGH. In 9 normal subjects, the urinary IRHGH ranged from 28–53 ng/24 h. The excretion of urinary IRHGH was increased in acromegaly and was diminished in some, but not in all patients with adult hypopituitarism. The urinary IRHGH was further studied by gel filtration. It was recovered in one peak corresponding to a molecular weight of approximately 20 000 – 30 000. However, in the present work it was not clarified whether the urinary IRHGH represents pituitary HGH excreted in the urine or a metabolite of high molecular weight with retained immunological properties.

scholarly journals Purification and properties of the hexosaminidase A-activating protein from human liver

1977 ◽  
Vol 167 (3) ◽  
pp. 693-701 ◽  
Author(s):  
Peter Hechtman ◽  
Dorothy LeBlanc
Keyword(s):  
Human Liver ◽  
Gel Filtration ◽  
Disc Electrophoresis ◽  
Exchange Chromatography ◽  
Hydrolase Activity ◽  
Gm2 Ganglioside ◽  
Purification And Properties ◽  
Hexosaminidase A ◽  
Hydrolysis Of

Human liver extracts contain an activating protein which is required for hexosaminidase A-catalysed hydrolysis of the N-acetylgalactosaminyl linkage of GM2 ganglioside [N-acetylgalactosaminyl-(N-acetylneuraminyl) galactosylglucosylceramide]. A partially purified preparation of human liver hexosaminidase A that is substantially free of GM2 ganglioside hydrolase activity is used to assay the activating protein. The proceudres of heat and alcohol denaturation, ion-exchange chromatography and gel filtration were used to purify the activating protein over 100-fold from crude human liver extracts. When the purified activating protein is analysed by polyacrylamide-gel disc electrophoresis, two closely migrating protein bands are seen. When purified activating protein is used to reconstitute the GM2 ganglioside hydrolase activity, the rate of reaction is proportional to the amount of hexosaminidase A used. The activation is specific for GM2 ganglioside and and hexosaminidase A. The activating protein did not stimulate hydrolysis of asialo-GM2 ganglioside by either hexosaminidase A or B. Hexosaminidase B did not catalyse hydrolysis of GM2 ganglioside with or without the activator. Kinetic experiments suggest the presence of an enzyme–activator complex. The dissociation constant of this complex is decreased when higher concentrations of substrate are used, suggesting the formation of a ternary complex between enzyme, activator and substrate. Determination of the molecular weight of the activating protein by gel-filtration and sedimentation-velocity methods gave values of 36000 and 39000 respectively.

The Sequential Bithermal Binaural Caloric Test: I. A Statistical Analysis of Normal Subject Responses

1973 ◽  
Vol 82 (3) ◽  
pp. 3-8 ◽  
Author(s):  
David D. Custer ◽  
Franklin O. Black ◽  
William G. Hemenway ◽  
John I. Thornby
Keyword(s):  
Statistical Analysis ◽  
Confidence Intervals ◽  
Normal Subjects ◽  
Significant Response ◽  
Caloric Test ◽  
Test Results ◽  
Normal Limits ◽  
Normal Subject

Statistical analysis of bithermal binaural caloric test results gave a statistically significant response difference between ears for right-handed normal subjects. There were no significant response differences due to temperature of stimulation or due to the interaction between temperature of stimulation and ear stimulated. Of the four different orders of stimulation tested, all were statistically equivalent. Because intrasubject variance from a statistical analysis of normal subject responses was evidently much smaller than percentage based variance, the use of absolute differences or confidence intervals for determination of normal limits should yield more precise normal criteria than percentage based comparisons allow.

scholarly journals Antibody-dependent lymphocyte-mediated granulocyte cytotoxicity in man

Blood ◽  
1978 ◽  
Vol 51 (1) ◽  
pp. 97-108 ◽  
Author(s):  
GL Logue ◽  
R Kurlander ◽  
P Pepe ◽  
W Davis ◽  
H Silberman
Keyword(s):  
Gradient Centrifugation ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Normal Subjects ◽  
Exchange Chromatography ◽  
Phagocytic Cells ◽  
Igg Antibody ◽  
Microtiter Plates ◽  
Granulocyte Antibodies

Abstract Sera from two patients with granulocytopenia associated with collagen vascular disease caused the destruction of normal human granulocytes by autologous lymphocytes in vitro. Granulocyte cytotoxicity was measured by the release of 51Cr during incubation with test sera and lymphocytes in microtiter plates. Between 8% and 46% granulocytoxicity was produced in granulocytes from 8 normal donors by the sera from these two patients. Less than 6% granulocytotoxicity was seen with the sera from 14 normal subjects and 29 patient controls. Treatment of lymphocyte preparations with carbonyl iron and magnetic separation to remove phagocytic cells or treatment with complement-coated red cells followed by repeated gradient centrifugation to remove complement receptor- bearing lymphocytes did not reduce the granulocytotoxicity. There was a dose-response relationship between the concentration of positive sera and granulocytotoxicity. When these sera were fractionated by Sephadex G-200 gel filtration and by ion-exchange chromatography with DEAE- cellulose, the active component appeared in the IgG-containing fractions. Thus, IgG antibody-dependent, lymphocyte-mediated granulocyte cytotoxicity represents a means of detecting human granulocyte antibodies and is a possible mechanism of autoimmune neutropenia in these two patients.

Determination of free follistatin levels in sera of normal subjects and patients with various diseases

1996 ◽  
Vol 135 (3) ◽  
pp. 345-351 ◽  
Author(s):  
Yukihiro Sakamoto ◽  
Yasumi Shintani ◽  
Kazuyo Harada ◽  
Masahiro Abe ◽  
Keiji Shitsukawa ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Gel Filtration ◽  
Normal Subjects ◽  
Activin A ◽  
Solid Cancer ◽  
Serum Samples ◽  
Response Curves ◽  
Standard Curve ◽  
Human Sera

Sakamoto Y. Shintani Y, Harada K, Abe M, Shitsukawa K, Saito S. Determination of free follistatin levels in sera of normal subjects and patients with various diseases. Eur J Endocrinol 1996:135:345–51. ISSN 0804–4643 We developed an assay system for measuring free follistatin by using an anti-follistatin mouse monoclonal antibody and [125I]activin A. The sensitivity of this assay was 0.5 μg/l and crossreactivities with inhibin. luteinizing hormone, follicle-stimulating hormone and growth hormone were all less than 0.5%. The dose-response curves of human sera and follicular fluid were parallel to the standard curve, and the follicular fluid contained a large amount of follistatin (6.4 ± 0.5 mg/l, mean ± SEM; N = 13). The within- and between-assay coefficients of variation calculated from the analysis of serum samples of four different concentrations were 3.3–7.8% and 3.9–11.0%, respectively. The recovery rates of free follistatin at five different doses were 86.4–102.4%. When activin A was added to the same sample, free follistatin recovery rate declined dose-dependently. Gel filtration analyses of human serum and follicular fluid resulted in a single peak corresponding to authentic follistatin. Using this assay, free follistatin concentrations in sera were measured in normal, pregnant and diseased subjects. The free follistatin level in serum of normal adults was 3.5 ± 0.2 μg/l (N = 60), which was significantly elevated in pregnant women (16.7 ± 1.3 μg/l, N = 56), and in patients with chronic liver disease (8.1 ± 1.1 μg/l, N = 20), chronic renal failure (6.7 ± 0.9 μg/l, N = 42), advanced solid cancer (8.5 ± 1.0 μg/l, N = 39) and hematological malignancies (6.8 ± 1.0 μg/l, N = 18). These data indicated that the free follistatin concentration in serum is detectable and varies during pregnancy and in various diseased states. Yukihiro Sakamoto, First Department of Internal Medicine, School of Medicine, The University of Tokushima, 3-18-15 Kuramoto-cho, Tokushima 770, Japan

scholarly journals Very-high-field n.m.r. studies of bovine lung heparan sulphate tetrasaccharides produced by nitrous acid deaminative cleavage. Determination of saccharide sequence, uronate composition and degrees of sulphation

1984 ◽  
Vol 223 (2) ◽  
pp. 495-505 ◽  
Author(s):  
P N Sanderson ◽  
T N Huckerby ◽  
I A Nieduszynski
Keyword(s):  
Nitrous Acid ◽  
High Field ◽  
General Structure ◽  
Heparan Sulphate ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Very High ◽  
Specific Sequences

Tetrasaccharides with the general structure UA-GlcNAc-GlcUA-aManOH (where UA represents uronate, GlcNAc N-acetylglucosamine, GlcUA glucuronate and aManOH anhydromannitol) were prepared from low-sulphated heparan sulphates of bovine lung origin by complete nitrous acid deaminative cleavage followed by reduction and fractionated by gel filtration. Ion-exchange chromatography of the tetrasaccharides yielded three major fractions in approximate yields of 37%, 45% and 14%. These were shown to be non-, mono- and di-sulphated respectively. Complete structural characterization of the tetrasaccharide fractions by quantitative high-field n.m.r. spectroscopy showed that each fraction contained only two discrete species and led to the following observations. (1) All of the uronate residues in the tetrasaccharides (and in larger oligosaccharides) are unsulphated, and hence sulphated iduronate [IdUA(2SO3)] must occur exclusively within -GlcNSO3-IdUA(2SO3)-GlcNSO3- sequences (where GlcNSO3 represents N-sulpho-glucosamine) in the parent polymers. (2) The GlcNAc residues in the tetrasaccharides are more highly C-6-O-sulphated than are the aManOH residues, and furthermore sulphation on the aManOH appears to occur only where the GlcNAc is also sulphated. (3) Where the GlcNAc is C-6-O-sulphated, iduronate is the major non-reducing terminal residue, whereas glucuronate predominates in this position if the GlcNAc is unsulphated. The quantitative data obtained are used to determine the degree of C-6-O-sulphation of glucosamine residues in specific sequences within the parent heparan sulphates.

Purification, Biochemical Characterization and Decolorization Efficiency of Laccases from Peach and Cherry Cultures of Pleutorus eryngii: A Comparative Study

2020 ◽  
Vol 27 (7) ◽  
pp. 623-634 ◽  
Author(s):  
Merve Akpinar ◽  
Raziye Ozturk Urek
Keyword(s):  
Biochemical Characterization ◽  
Gel Filtration ◽  
Industrial Effluents ◽  
Anion Exchange Chromatography ◽  
Biochemical Properties ◽  
Exchange Chromatography ◽  
Kinetic Characteristics ◽  
Chemical Agents ◽  
Decolorization Efficiency

Background:: Laccases (Lacs) are used potentially in industrial and biotechnological applications such as decolorization of dyes, degradation of industrial effluents, delignification, etc. thanks to their large varieties of substrate specificities and excellent catalytic efficiencies. The efficient utilizations of Lacs in these applications mostly depend on the identifying their biochemical properties. Objective: The goal of this research is to investigate the purification, biochemical characterization and decolorization efficiencies of Lacs. Methods: Pleurotus eryngii was incubated on peach (PC) and cherry (CC) wastes under optimized solid state fermentation conditions. Then, the enzymes extracts were purified by ammonium sulfate precipitation, anion exchange chromatography, gel filtration, respectively. Lacs fractions were subjected to electrophoretic analyses as well as their structural and kinetic characteristics. Also, the effects of selected chemical agents on purified Lacs activities and determination of decolorization efficiencies were studied. Results: As the results of purification processes of Lacs from both cultures, 3.94-fold purification was obtained for PC, while it was 5.34 for CC. The electrophoretic results of purified Lacs illustrated the single bands of protein (30±1 kDa) in accordance with the results after gel filtration. The Km values of Lacs from PC and CC were respectively detected as 1.1381 and 0.329 mM for ABTS. The selected agents partially/completely inhibited Lac activities. The highest decolorization efficiencies of purified Lacs from PC and CC were separately obtained as 53 and 11.8%. Conclusion: The results clearly indicated that the performances of Lacs from both cultures in decolorization application are different from each other depending their activities, biochemical and kinetic characteristics.

The Sequential Bithermal Binaural Caloric Test

1973 ◽  
Vol 82 (6_suppl) ◽  
pp. 3-8
Keyword(s):  
Statistical Analysis ◽  
Confidence Intervals ◽  
Normal Subjects ◽  
Significant Response ◽  
Caloric Test ◽  
Test Results ◽  
Normal Limits ◽  
Normal Subject

Statistical analysis of bithermal binaural caloric test results gave a statistically significant response difference between ears for right-handed normal subjects. There were no significant response differences due to temperature of stimulation or due to the interaction between temperature of stimulation and ear stimulated. Of the four different orders of stimulation tested, all were statistically equivalent. Because intrasubject variance from a statistical analysis of normal subject responses was evidently much smaller than percentage based variance, the use of absolute differences or confidence intervals for determination of normal limits should yield more precise normal criteria than percentage based comparisons allow.

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