RADIOIMMUNOASSAY OF TETRAHYDROCORTICOSTERONE (THB) IN HUMAN URINE

1978 ◽  
Vol 88 (1) ◽  
pp. 139-148 ◽  
Author(s):  
K.-H. Kohl ◽  
K.-H. Gless ◽  
S. Abdelhamid ◽  
B. Penke ◽  
P. Vecsei
Keyword(s):  
New Zealand ◽  
Bovine Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Normal Values ◽  
Chromatographic System ◽  
Acth Stimulation ◽  
Cross Reactions ◽  
Dexamethasone Suppression ◽  
New Zealand Rabbits

ABSTRACT Antisera against tetrahydrocorticosterone (THB) (one of the metabolites of the glucocorticoid corticosterone) were raised in white New Zealand rabbits. A 3α,5β-THB-20-oxime-bovine serum albumin complex was used as antigen. The resulting titers were 1/300–1/19 000. The cross-reactions of the best antiserum were: THDOC 2.25%, THB-21-glucuronide 39%, and THF-21-glucuronide 4.8%. Other steroids and steroid metabolites have given negligible cross-reactions, e. g. allo-(3α,5α-)-THB 0.53 %, tetrahydro-11-dehydro-corticosterone (THA) 0.92%, allo-THA 0%, and THB-3-glucuronide 0 %. Two methods were introduced: a) for screening an estimation of THB-glucuronide, mostly THB-21-glucuronide ("THB-gluc."), in unprocessed urine after simple dilution b) specific estimation of THB using one paper chromatographic system. The values estimated by method a) showed a highly significant correlation (r = 0.9654, P < 0.01) with the values obtained by technique b). Normal values (method b) measured in subjects hospitalized under standardized conditions were: 21.8–177.6; mean 89.1 μg/24 h. THB levels and "THB-gluc." levels react to ACTH stimulation and dexamethasone suppression and thus we propose that the methods are useful for the diagnosis of hyper- and hypocorticosterone production states.

RADIOIMMUNOASSAYS OF TETRAHYDROCORTISONE AND TETRAHYDROCORTISOL IN HUMAN URINE

1977 ◽  
Vol 86 (2) ◽  
pp. 369-379 ◽  
Author(s):  
H. Will ◽  
R. Aderjan ◽  
Th. Winkler ◽  
B. Penke ◽  
P. Vecsei
Keyword(s):  
New Zealand ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Secretion Rate ◽  
Cortisol Secretion ◽  
Cross Reactions ◽  
New Zealand Rabbits

ABSTRACT A simple radioimmunoassay for the determination of tetrahydrocortisone (THE)1), and tetrahydrocortisol (THF) as well as their 21-glucosiduronate conjugates is described. White New Zealand rabbits were immunized with THF- and THE-20-oxime-bovine serum-albumin conjugates. The resulting antisera proved to be very specific. Both allo-THE and allo-THF gave minimal cross-reactions; however, THE- and THF-21-glucosiduronate bind with the corresponding antibodies at a level of 100 %. The following types of estimations were introduced: a) Estimation of the sum of unconjugated THF and THF-21-glucosiduronate in highly diluted unprocessed human urine. b) A similar estimation of THE and THF-21-glucosiduronate as in a). c) Estimation of THF after β-glucuronidase treatment and dilution without further procedures. d) A similar estimation for THE as in c). THF and THE values correlate significantly with known parameters that characterize the cortisol production, such as cortisol secretion rate and the urinary corticoid (C21-α-ketolic-steroid) levels.

RADIOIMMUNOASSAYS OF TETRAHYDROALDOSTERONE (TH-ALDO) IN HUMAN URINE

1978 ◽  
Vol 87 (3) ◽  
pp. 596-608 ◽  
Author(s):  
K.-H. Kohl ◽  
P. Vecsei ◽  
S. Abdelhamid
Keyword(s):  
Bovine Serum Albumin ◽  
Normal Values ◽  
Rapid Screening ◽  
Chromatographic System ◽  
Polar Material ◽  
Butanol Extract ◽  
Specific Antisera ◽  
Aldosterone Production ◽  
Rapid Screening Test ◽  
New Zealand Rabbits

ABSTRACT Specific antisera against tetrahydroaldosterone (TH-Aldo) were raised in two white New Zealand rabbits. 3α,5β-TH-aldo-20-oxime-bovine-serum albumin complex was used as antigen. The resulting titers were 1:18 000 and 1:16 000. Except tetrahydrocortisol (THF) (0.23%) and tetrahydro-18-hydroxy-11-dehydrocorticosterone (18-OH-THA) (3.2 %), all steroids and steroid metabolites gave negligible cross-reactions. Immunograms of the paper chromatograms made from the n-butanol-extract of the urines, as well as after β-glucuronidase treatment and dichlormethane extraction, were studied to further define the specificity of the antiserum. Antibody H1 (used in this study) reacted with aldosterone-18-gluc., a TH-aldosterone-glucuronide (probably the 21-glucuronide) and an unidentified less polar material. Two methods were developed: a) TH-Aldo-glucuronidc(s) estimation after ethylacetate pre-extraction as a rapid screening test of endogenous aldosterone production. b) estimation of TH-aldosterone using one chromatographic system. The results of method a) showed a significant correlation with the values obtained by technique b). Normal values (method b) were 25.88 ± 16.50 μg/24h (range 9.5–64.8 μg/24h). A significant correlation was also shown between the TH-aldo (technique b) and 18-gluc. values.

scholarly journals Effects of weak bases on the degradation of endogenous and exogenous proteins by rat yolk sacs

1980 ◽  
Vol 188 (3) ◽  
pp. 895-903 ◽  
Author(s):  
G Livesey ◽  
K E Williams ◽  
S E Knowles ◽  
F J Ballard
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Normal Values ◽  
Free Medium ◽  
Weak Base ◽  
Weak Bases ◽  
Endogenous Proteins ◽  
Hydrolysis Of

In rat yolk sacs incubated in vitro, the rates of degradation of endogenous [3H]leucine-labelled proteins and of pinocytically ingested 125I-labelled bovine serum albumin were both decreased in the presence of either ammonium, methylammonium or ethylammonium ions (0-20 mM) or much lower concentrations of chloroquine (0-500 microM). These effects were also accompanied by an inhibition of pinocytosis, as measured by the rate of uptake of 125I-labelled polyvinylpyrrolidone, and by a fall in the [ATP]/[ADP] ratio within the tissue. Re-incubation in inhibitor-free medium of yolk sacs previously exposed to a weak base restored pinocytic and proteolytic capacities, except for tissues exposed to chloroquine at concentrations above 0.1 mM (these appeared to be cytotoxic); an attendent rise in [ATP]/[ADP] ratios to near normal values was also observed. Weak bases, at concentrations that fully arrested the breakdown of 125I-labelled albumin, failed to inhibit by more than 45% the degradation of [3H]leucine-labelled endogenous proteins. Since 125I-labelled bovine serum albumin has been shown to be degraded entirely intralysosomally by yolk sacs, this suggests either that the hydrolysis of endogenous proteins is shared between lysosomes and some other site or that, unlike 125I-labelled albumin, some endogenous proteins can be degraded within lysosomes at abnormally high pH.

scholarly journals A modified radioimmunoassay of aldosterone

1995 ◽  
Vol 41 (2) ◽  
pp. 33-34
Author(s):  
N. P. Goncharov ◽  
G. V. Katsiya ◽  
V. Yu. Butnev ◽  
L. B. Khagundokova
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Adrenal Cortex ◽  
Bovine Serum ◽  
High Specificity ◽  
New Method ◽  
Isolation And Purification ◽  
Cross Reactions ◽  
Chromatographic Isolation

A method of radioimmunoassay of aldosterone in the blood of humans and monkeys has been developed. Antiserum was prepared on rabbits against aldosterone-3-(0-carboxymethyloxime) conjugate with bovine serum albumin. The specificity of antiserum was tested in cross-reactions with 32 related steroids. High specificity of the antiserum in this method helped measure aldosterone levels without preliminary chromatographic isolation and purification of this compound. The sensitivity of the new method is 12.5 pg per 0.1 ml. Reproducibility of the method between different series of measurements was no more than 15%, within the same series below 12%. The method was used to assess the mineralocorticoid function of the adrenal cortex of man and monkeys.

scholarly journals An enzyme-linked immunoassay for the collagen cross-link pyridinoline

1982 ◽  
Vol 207 (3) ◽  
pp. 617-620 ◽  
Author(s):  
S P Robins
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Urine Samples ◽  
Cross Link ◽  
Human Urine Samples

An inhibition immunoassay method for the determination of pyridinoline was developed with the use of microtitre plates coated with a pyridinoline-gelatin conjugate and rabbit antisera directed against pyridinoline linked to bovine serum albumin. The sensitivity of the assay is about 2pmol of pyridinoline, and the presence of related pyridinium and lysine-derived compounds does not significantly interfere with the procedure. Its application to tissue and human urine samples is described.

Protein-protected red emittive copper nanoclusters as a fluorometric probe for highly sensitive biosensing of creatinine

2018 ◽  
Vol 10 (29) ◽  
pp. 3666-3674 ◽  
Author(s):  
Ramar Rajamanikandan ◽  
Malaichamy Ilanchelian
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Urine Samples ◽  
Copper Nanoclusters ◽  
Human Urine Samples ◽  
Fluorescent Nanoprobe ◽  
Highly Sensitive ◽  
Optical Recognition

We describe red emittive bovine serum albumin-modified copper nanoclusters (BSA-CuNCs) applied as a precise fluorescent nanoprobe for the optical recognition of creatinine in human urine samples.

scholarly journals Recommendations for the detection of Leptospira in urine by PCR

2004 ◽  
Vol 37 (2) ◽  
pp. 131-134 ◽  
Author(s):  
Paula M.A. Lucchesi ◽  
Guillermo H. Arroyo ◽  
Analía I. Etcheverría ◽  
Alberto E. Parma ◽  
Alfredo C. Seijo
Keyword(s):  
Bovine Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Room Temperature ◽  
Freezing And Thawing ◽  
Sample Processing ◽  
Negative Results ◽  
Inhibitory Compounds ◽  
Increased Sensitivity ◽  
Acidic Urine

In the present study PCR was applied to detect leptospires in human urine. Several approaches for sample processing were evaluated to optimize the detection of leptospires in urine mixed with this bacterium. Furthermore, some changes in the composition of the reaction mix were studied. No amplification was observed in acidic urine, therefore neutralization of the sample immediately after collection is strongly recommended. PBS gave better results than Tris or NaOH as neutralizing reagents. Freezing and thawing of samples before processing yielded negative results. Elimination of epithelial cells, leukocytes and crystals by centrifugation at 3,000 rpm at room temperature increased sensitivity. In addition, both the washing step after collecting leptospires by centrifugation and the inclusion of 0.1% bovine serum albumin in the reaction mix minimized the interference of other inhibitory compounds. These modifications were useful to improve the detection of Leptospira in urine by PCR.

Effects of blood-free and protein-free perfusion on CFC in the isolated cat hindlimb

1983 ◽  
Vol 245 (6) ◽  
pp. H911-H919 ◽  
Author(s):  
P. D. Watson
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Water Flow ◽  
Electrolyte Solution ◽  
Bovine Serum ◽  
Normal Values ◽  
Filtration Coefficient ◽  
Constant Flow ◽  
Capillary Filtration ◽  
Capillary Filtration Coefficient

To investigate the effects of blood and protein on capillary filtration coefficient (CFC), isolated cat hindlimbs were perfused at constant flow from a reservoir containing one of three perfusates: 1) an electrolyte solution also used for dialyzing bovine serum albumin (BSA) and hence labeled dialysate, 2) dialyzed 6.5–7.0 g/100 ml BSA, labeled albumin, and 3) albumin plus cat blood, labeled control. CFC during control perfusion averaged 0.0137 +/- 0.003 (SD, n = 26) and increased slowly with time. CFCs with dialysate and albumin were 3.1 +/- 0.56 (n = 11) and 1.7 +/- 0.52 (n = 15) times control, respectively. However, the ratio in a particular experiment depended on control CFC, the ratio being low when the control CFC was low and high when control CFC was high. The data can be explained if water flow occurs through two types of transcapillary pathways, one having a hydraulic capacity constant from animal to animal and insensitive to blood and protein, the second varying in hydraulic capacity and requiring blood and protein for its normal values.

Localization of Intracellular Antigens in thin Sections with Ferritin Labeled Antibody

1970 ◽  
Vol 28 ◽  
pp. 174-175
Author(s):  
M.S. Shahrabadi ◽  
T. Yamamoto
Keyword(s):  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Antigenic Determinants ◽  
Conjugate Prior ◽  
Thin Sections ◽  
Specific Attachment ◽  
Intracellular Antigen ◽  
Antigen Antibody ◽  
Ideal Method ◽  
The Ideal

The technique of labeling of macromolecules with ferritin conjugated antibody has been successfully used for extracellular antigen by means of staining the specimen with conjugate prior to fixation and embedding. However, the ideal method to determine the location of intracellular antigen would be to do the antigen-antibody reaction in thin sections. This technique contains inherent problems such as the destruction of antigenic determinants during fixation or embedding and the non-specific attachment of conjugate to the embedding media. Certain embedding media such as polyampholytes (2) or cross-linked bovine serum albumin (3) have been introduced to overcome some of these problems.

Improvement of an artificial test substrate technique for evaluation of em immunocytochemical labeling

1987 ◽  
Vol 45 ◽  
pp. 762-763
Author(s):  
G. D. Gagne ◽  
M. F. Miller
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Artificial Substrate ◽  
Chemical Fixation ◽  
As If

We recently described an artificial substrate system which could be used to optimize labeling parameters in EM immunocytochemistry (ICC). The system utilizes blocks of glutaraldehyde polymerized bovine serum albumin (BSA) into which an antigen is incorporated by a soaking procedure. The resulting antigen impregnated blocks can then be fixed and embedded as if they are pieces of tissue and the effects of fixation, embedding and other parameters on the ability of incorporated antigen to be immunocyto-chemically labeled can then be assessed. In developing this system further, we discovered that the BSA substrate can also be dried and then sectioned for immunolabeling with or without prior chemical fixation and without exposing the antigen to embedding reagents. The effects of fixation and embedding protocols can thus be evaluated separately.

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