Time-resolved immunofluorometric assay of 17 beta-hydroxysteroid dehydrogenase in plasma

1991 ◽  
Vol 37 (8) ◽  
pp. 1412-1415 ◽  
Author(s):  
O Mäentausta ◽  
M Menjivar ◽  
R Vihko
Keyword(s):  
Polyclonal Antibodies ◽  
Endometrial Adenocarcinoma ◽  
Plasma Concentrations ◽  
Hydroxysteroid Dehydrogenase ◽  
Minimum Detectable Concentration ◽  
Antibody Molecule ◽  
Time Resolved ◽  
Detectable Concentration ◽  
The Mean ◽  
Mean Concentration

Abstract We describe a time-resolved immunofluorometric assay (TR-IFMA) for human 17 beta-hydroxysteroid dehydrogenase (17HSD) in which antibody-coated microtiter strip wells and europium chelate-labeled polyclonal antibodies are used. In preparing the label, a polyclonal antibody is affinity-purified and derivatized with diethylenetriamine-pentaacetic acid. With this derivative, five to eight europium ions can be combined with one antibody molecule without decreasing the antibody's immunoreactivity. The minimum detectable concentration of 17HSD is 0.13 microgram/L; the intra- and interassay CVs are less than 8% and less than 15%, respectively, for concentrations between 0.3 and 100 micrograms/L. There is no difference between the concentrations of 17HSD in plasma specimens taken during the proliferative and luteal phases of the menstrual cycle, the measured mean concentration being 0.22 microgram/L. We found no correlation between plasma 17HSD and progesterone concentrations. The plasma concentrations of 17HSD increase during pregnancy, the mean concentrations being 1.5, 4.4, and 12.5 micrograms/L, during the first, second, and third trimesters of pregnancy, respectively. In the specimens from 18 men, the mean concentration was 0.18 microgram/L. In six plasma specimens from patients with endometrial adenocarcinoma, the mean concentration was 0.20 micrograms/L. Pre-analytical aspects are important in the assay of 17HSD because of the lability of the enzyme protein. Preferably, blood should be sampled into EDTA-containing tubes, plasma should be separated within 15 min, and glycerol must be added without delay to a final volume of 200 mL/L.

Time-resolved immunofluorometric assay of human prostate-specific antigen

1990 ◽  
Vol 36 (1) ◽  
pp. 92-95 ◽  
Author(s):  
P Vihko ◽  
R Kurkela ◽  
J Ramberg ◽  
I Pelkonen ◽  
R Vihko
Keyword(s):  
Human Serum ◽  
Prostate Specific Antigen ◽  
Prostatic Cancer ◽  
Specific Antigen ◽  
Prostatic Hyperplasia ◽  
Minimum Detectable Concentration ◽  
Antibody Molecule ◽  
Time Resolved ◽  
Radioactive Label ◽  
Detectable Concentration

Abstract Assay of human serum prostate-specific antigen (PSA) is gaining importance in diagnosis and follow-up of prostatic cancer. In this time-resolved immunofluorometric assay of PSA, strip-wells were coated with a polyclonal antibody against PSA. To prepare the label, a monoclonal antibody displaying high affinity towards PSA was purified and derivatized with diethylenetriaminepentaacetic acid. With use of this derivative, seven to eight Eu atoms could be combined with one antibody molecule with no decrease in immunoreactivity. The minimum detectable concentration of PSA was 0.12 microgram/L. In 60 of 63 women studied, the PSA concentration in serum was less than 0.2 microgram/L. The increase in PSA in serum of asymptomatic men 51 years old or older, as compared with that for younger subjects, was possibly a result of "occult" prostatic hyperplasia. Most of the patients with prostatic hyperplasia or prostatic cancer had higher PSA concentrations than did subjects of ages less than 50 years or asymptomatic age-matched healthy subjects. Results compared favorably with those by an established technique relying on the use of radioactive label. Our method for measuring PSA in human serum is convenient, inexpensive, and well compatible with present clinical practice.

scholarly journals Intact transferrin receptors in human plasma and their relation to erythropoiesis

Blood ◽  
1990 ◽  
Vol 75 (1) ◽  
pp. 102-107 ◽  
Author(s):  
HA Huebers ◽  
Y Beguin ◽  
P Pootrakul ◽  
D Einspahr ◽  
CA Finch
Keyword(s):  
Human Plasma ◽  
Polyclonal Antibodies ◽  
Close Correlation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transferrin Receptors ◽  
Erythroid Hyperplasia ◽  
The Mean ◽  
Tissue Receptors ◽  
Mean Concentration ◽  
Normal Adults

Abstract Intact transferrin receptor molecules complexed with transferrin were found in human plasma. The concentration of receptors was determined by an enzyme-linked immunosorbent assay that uses polyclonal antibodies. The mean concentration of 8,279 micrograms/L in 56 normal adults appears to be unrelated to age or sex. Additional receptor measurements were performed on plasmas from 260 subjects with erythropoietic disorders. Decreased concentration of plasma receptors was found in patients with erythroid hypoplasia and increased numbers in those with erythroid hyperplasia. Ferrokinetic measurements of erythropoiesis were compared with numbers of receptors in 148 subjects, and a close correlation was found (r = .86). Both sets of values, measured in different conditions and expressed in relation to normal, were consistent with expected values. Receptor values were unproportionally increased only in conditions of iron deficiency. It is concluded that plasma receptors have a constant relationship to tissue receptors, and their number in most instances reflects the rate of erythropoiesis.

Immunofluorometric demonstration and quantification of placental protein 5 in the absence of pregnancy.

1988 ◽  
Vol 34 (8) ◽  
pp. 1591-1593 ◽  
Author(s):  
R Bützow ◽  
H Alfthan ◽  
U H Stenman ◽  
A M Suikkari ◽  
H Bohn ◽  
...  
Keyword(s):  
Detection Limit ◽  
Solid Phase ◽  
Polyclonal Antiserum ◽  
Serum Samples ◽  
Measuring Range ◽  
Time Resolved ◽  
Healthy Men ◽  
Placental Protein ◽  
The Mean ◽  
Mean Concentration

Abstract This time-resolved immunofluorometric assay (IFMA) developed for measurement of placental protein 5 (PP5) involves two antibodies: a monoclonal anti-PP5 antibody attached to a solid phase and an europium(III) chelate-labeled polyclonal anti-PP5 antibody as a tracer. The measuring range is 0.05-100 micrograms/L and the detection limit is 20 times lower than that of a PP5 radioimmunoassay (RIA) performed with the same polyclonal antiserum. By IFMA, PP5 could be detected and quantified in all plasma and serum samples of nonpregnant and pregnant individuals, whereas PP5 was undetectable by RIA in serum of healthy men and nonpregnant women. The mean concentration of PP5 in sera from men was 0.43 micrograms/L (SD 0.13, range 0.19-0.75, n = 47) and in sera from nonpregnant women 0.49 micrograms/L (SD 0.19, range 0.20-0.90, n = 41). PP5 concentrations in serum showed no systematic variation during the menstrual cycle. In serum samples from 60 pregnant women the results obtained by IFMA and RIA correlated well (r = 0.97).

scholarly journals Plasma and ovarian oestradiol and the variability in the LH surge induced in ewes by the ram effect

Reproduction ◽  
2015 ◽  
Vol 149 (5) ◽  
pp. 511-521 ◽  
Author(s):  
Claude Fabre-Nys ◽  
Audrey Chanvallon ◽  
Nathalie Debus ◽  
Dominique François ◽  
Frédéric Bouvier ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Plasma Concentrations ◽  
Feedback Signal ◽  
Sexually Active ◽  
Lh Surge ◽  
Before And After ◽  
Ram Effect ◽  
The Mean ◽  
Mean Concentration

The proportion of anoestrous ewes ovulating after exposure to a sexually active ram is variable mainly due to whether an LH surge is induced. The aim of this study was to determine the role of oestradiol (E2) in the ram-induced LH surge. In one study, we measured the plasma concentrations of E2 in ewes of different breeds before and after the ‘ram effect’ and related these patterns to the presence and latency of the LH surge, while another compared ovarian responses with the ‘ram effect’ following exposure to rams for 2 or 12 h. In all ewes, the concentration of E2 increased 2–4 h after rams were introduced and remained elevated for 14.5±0.86 h. The quantity of E2 secreted before the LH surge varied among breeds as did the mean concentration of E2. The granulosa cells of IF ewes collected after 12 h exposure to rams secreted more E2 and progesterone and had higher levels of StAR than the 2 h group but in MV ewes there was no differences between these groups for any of these parameters. These results demonstrate that the LH surge induced by the rams is a result of increased E2 secretion associated with increased levels of STAR in granulosa cells and that these responses varied among breeds. The results suggest that the variable occurrence of a LH surge and ovulation may be the result of variable ovarian responses to the ‘ram effect’ and insensitivity of the hypothalamus to the E2-positive feedback signal.Free French abstract: A French translation of this abstract is freely available at http://www.reproduction-online.org/content/149/5/511/suppl/DC1.

Plasma oestradiol-17β and testosterone concentrations as possible causes of the infertility of congenitally obese Zucker rats

1983 ◽  
Vol 99 (3) ◽  
pp. 485-490 ◽  
Author(s):  
E. M. Whitaker ◽  
M. A. Shaw ◽  
G. R. Hervey
Keyword(s):  
Plasma Concentrations ◽  
Gonadal Hormones ◽  
Oestrous Cycle ◽  
Zucker Rats ◽  
Obese Female ◽  
Obese Rats ◽  
Obese Zucker Rats ◽  
The Mean ◽  
Plasma Oestradiol ◽  
Mean Concentration

The plasma oestradiol-17β concentrations of obese and non-obese female Zucker rats have been measured in three phases of the oestrous cycle. The oestradiol concentrations of both phenotypes were similar, and changed normally with the oestrous cycle. The weights of the uteri also changed normally with the cycle. Plasma androgen concentrations in male Zucker rats have also been measured: the mean concentration was slightly but significantly lower in obese rats, and androgen-sensitive tissues were slightly reduced in weight. The oestradiol-17β concentrations in males of both phenotypes were similar. It seems unlikely that deficient plasma concentrations of gonadal hormones cause the infertility of obese rats of either sex.

VARIATIONS IN PLASMA CONCENTRATIONS OF VASOPRESSIN DURING THE MENSTRUAL CYCLE

1981 ◽  
Vol 89 (2) ◽  
pp. 263-266 ◽  
Author(s):  
MARY L. FORSLING ◽  
M. ÅKERLUND ◽  
P. STRÖMBERG
Keyword(s):  
Menstrual Cycle ◽  
Plasma Concentrations ◽  
Hormonal Changes ◽  
Vasopressin Release ◽  
Healthy Women ◽  
Plasma Vasopressin ◽  
The Mean ◽  
Mean Concentration

Plasma vasopressin concentrations, determined by radioimmunoassay, were followed throughout the menstrual cycle in eight healthy women. The concentrations were found to depend on the day of the menstrual cycle. The mean concentration on day 1 was 0·5±0·08 (s.e.m.) μu./ml, while that on days 16–18 was 1·1±0·16 μu./ml. These values were significantly (P <0·02) different. Vasopressin release in women may thus depend on the hormonal changes during the menstrual cycle.

Is hirsutism an evolving syndrome?

1983 ◽  
Vol 97 (3) ◽  
pp. 379-387 ◽  
Author(s):  
Vincenzo Toscano ◽  
M. V. Adamo ◽  
Stefania Caiola ◽  
Sonia Foli ◽  
Elisa Petrangeli ◽  
...  
Keyword(s):  
Plasma Concentrations ◽  
Static Condition ◽  
Dehydroepiandrosterone Sulphate ◽  
Acth Stimulation ◽  
Group V ◽  
Peripheral Plasma ◽  
Group I ◽  
Menstruating Women ◽  
The Mean ◽  
Mean Concentration

The possibility that hirsutism is an evolving syndrome rather than a static condition involving only one gland has been considered. To assess this proposal 60 untreated hirsute patients aged 12–32 years were divided into five groups according to the duration of the hirsutism (< 1, 1–2, 2–3, 3–5 and > 5 years). Peripheral plasma concentrations of LH and FSH, androstenedione, dehydroepiandrosterone sulphate, testosterone, 5α-dihydrotestosterone, 5α-androstane-3α, 17β-diol, 5α-androstane-3β, 17β-diol, cortisol, oestradiol-17β and oestrone were determined by radioimmunoassay. When the values obtained were compared with those from normal menstruating women, the results showed that in group I there was a significant increase only in the mean plasma 5α-androstane-3α, 17β-diol concentration. The mean concentration of this steroid was also raised in all other groups. In groups II and III mean basal levels of plasma dehydroepiandrosterone sulphate were also significantly increased and showed a marked increase after ACTH stimulation (1 mg tetracosactide acetate, i.m.) as did the concentrations of androstenedione and 17α-hydroxyprogesterone. Finally, in groups IV and V, a significant increase in mean plasma concentrations of LH, androstenedione, oestrone and testosterone was found in the basal condition. The clinical picture also became gradually more severe from group I to group V. These data suggest that hirsutism could be an evolving syndrome progressively involving peripheral androgen metabolism, the adrenal gland and finally the ovary possibly through alterations of hypothalamic-pituitary function.

scholarly journals Validation of FFQ-based assessment of dietary lignans compared with serum enterolactone in Swedish women

2012 ◽  
Vol 109 (10) ◽  
pp. 1873-1880 ◽  
Author(s):  
Yulan Lin ◽  
Alicja Wolk ◽  
Niclas Håkansson ◽  
Jose Luis Peñalvo ◽  
Jesper Lagergren ◽  
...  
Keyword(s):  
Dietary Intake ◽  
Main Product ◽  
Human Subjects ◽  
Average Energy ◽  
Serum Concentrations ◽  
Time Resolved ◽  
Food Items ◽  
The Mean ◽  
Mean Concentration ◽  
Better Than

The validity of using FFQ to assess dietary lignans is uncertain. We aimed to validate the use of FFQ for the assessment of dietary intake of lignans compared to the serum biomarker enterolactone, the main product of dietary lignans' metabolism in human subjects. A random sample of women, aged 55–75 years, from the Swedish Mammography Cohort was selected. Information from two FFQ, the FFQ-87 (sixty-seven food items) and the FFQ-97 (ninety-three food items), and blood samples were collected. Dietary intake of lignans (secoisolariciresinol, matairesinol, lariciresinol, pinoresinol, medioresinol and syringaresinol) was assessed by the FFQ. Serum concentrations of enterolactone were analysed by time-resolved fluoroimmunoassay. The correlation coefficient between energy-adjusted lignan intake and serum enterolactone was estimated in crude and multivariable-adjusted models, taking into account the factors potentially influencing the serum enterolactone. Among the 135 participants aged 55–75 years, with a mean BMI of 26·7 kg/m2, the average energy-adjusted intake of total lignans was 1616 (sd 424) and 1516 (sd 409) μg/d according to the FFQ-87 (forty-five food items containing lignans) and the FFQ-97 (sixty-five food items containing lignans), respectively. The mean concentration of serum enterolactone was 23·2 (sd 15·4) nmol/l. The adjusted Pearson's correlation between dietary intake of lignans assessed by the FFQ-97 and serum enterolactone was statistically significant (r 0·22, P= 0·01). No significant correlation was observed for the FFQ-87 (r 0·09, P= 0·30). The present study indicates that the FFQ-97 might be better than the FFQ-87 for assessing dietary intake of lignans, although the correlation was low.

Plasma concentrations of progesterone, oestradiol-17β and 13,14-dihydro-15-oxo-prostaglandin F2α in the pregnant wallaby (Macropus rufogriseus rufogriseus)

1983 ◽  
Vol 97 (3) ◽  
pp. 369-377 ◽  
Author(s):  
M. T. Walker ◽  
R. T. Gemmell
Keyword(s):  
Plasma Concentrations ◽  
Prostaglandin F2α ◽  
Maternal Plasma ◽  
Gestation Length ◽  
Birth Process ◽  
Plasma Progesterone ◽  
Prostaglandin F ◽  
The Mean ◽  
Uterine Secretions ◽  
Mean Concentration

The concentrations of progesterone and oestradiol-17β in the maternal plasma of Bennett's wallaby, Macropus rufogriseus rufogriseus, were measured daily throughout gestation after reactivation of the diapausing corpus luteum by removal of the suckling pouch young (RPY). Progesterone increased from mean concentrations of 382–424 pmol/l (120–133 pg/ml) during lactation to reach peak concentrations of 908 ± 172 (s.e.m.) pmol/l (285 ± 54 pg/ml) (n = 8) 4 days after RPY and 971 ± 220 and 971 ± 229 pmol/l (305 ± 69 and 305 ± 72 pg/ml) (n = 7) 24 and 25 days after RPY respectively. The mean gestation length (RPY to birth) was 26·8 ± 0·6 (s.d.) days (n = 6, range 25·75–27·50 days). Immediately after birth the plasma progesterone concentration declined to 299 ± 51 (s.e.m.) pmol/l (94 ± 16 pg/ml) (n=6). Oestradiol-17β increased from mean concentrations of 291–553 pmol/l (80–152 pg/ml) during lactation to reach a peak concentration of 967 ± 331 pmol/l (266 ± 91 pg/ml) (n = 9) 1 day after RPY. The concentration declined from 7 days after RPY and fluctuated between mean concentrations of 273 and 480 pmol/l before reaching a minimum of 207 ± 69 pmol/l (57 ± 19 pg/ml) (n = 6) 19 days after RPY. A transient increase to 542 ± 207 pmol/l (n = 7) occurred at 22 days after RPY. Plasma concentrations declined to a low of 156 ± 55 pmol/l (43 ± 15 pg/ml) (n = 6) 5 days after parturition. The mean concentration of plasma 13,14-dihydro-15-oxo-prostaglandin F2α was less than 2·8 nmol/l (1 ng/ml) for all samples from 13 days after RPY until 4 days after parturition. The results suggest that oestradiol-17β may be important in the early stages of blastocyst reactivation to synergize with progesterone in stimulating uterine secretions. 13,14-Dihydro-15-oxo-prostaglandin F2α is unlikely to be involved in the birth process and any luteolytic effect is likely to be from a local production of PGF2α.

scholarly journals Time-resolved fluoroimmunoassay for bactericidal/permeability-increasing protein

1996 ◽  
Vol 5 (1) ◽  
pp. 47-50 ◽  
Author(s):  
J.-O. Häggblom ◽  
A. B. Jokilammi-Siltanen ◽  
H. Peuravuori ◽  
T. J. Nevalainen
Keyword(s):  
Human Serum ◽  
Polymorphonuclear Leukocytes ◽  
Antimicrobial Protein ◽  
Plasma Samples ◽  
Serum Samples ◽  
Time Resolved ◽  
Reactive Protein ◽  
The Mean ◽  
The Difference ◽  
Mean Concentration

Bactericidal/permeability-increasing protein (BPI) is a cationic antimicrobial protein produced by polymorphonuclear leukocytes, that specifically interacts with and kills Gram-negative bacteria. BPl competes with lipopolysaccharide-binding protein (LBP) secreted by liver cells into blood plasma for binding to lipopolysaccharide (LPS) and thus reduces the proinflammatory effects of LPS. We have developed a time-resolved fluoroimmunoassay for BPI and measured the concentration of BPI in human serum and plasma samples. The assay is based on a rabbit antibody against recombinant BPI. This antibody specifically adheres to polymorphonuclear leukocytes in immunostained human tissues. The difference in the serum concentration of BPI between unselected hospitalized patients with and without an infection was statistically significant. The mean concentration of BPI in serum samples was 28.3 μg/l (range 1.64–132, S.D. 26.8,n= 83). In contrast, there was no difference between the two groups in the BPI levels in plasma samples. For all individuals tested, BPI levels were consistently higher in plasma samples compared to the matched serum samples. The mean concentration of BPI in plasma samples was 52.3 μg/l (range 0.9–403, S.D. 60.6,n= 90). There was a positive correlation between the concentration of BPI and the white blood cell count as well as between the BPI concentration and C-reactive protein (CRP) in serum samples. In conclusion, the present study demonstrates that BPI can be quantified reliably by time-resolved fluoroimmunoassay in human serum samples.

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