Relaxin-induced changes in adenosine 3',5'-monophosphate levels in the human cervix

1985 ◽  
Vol 109 (1) ◽  
pp. 122-125 ◽  
Author(s):  
Anders Norström ◽  
Ingrid Wiqvist
Keyword(s):  
Pregnant Women ◽  
Needle Biopsy ◽  
Binding Assay ◽  
First Trimester ◽  
Cervical Tissue ◽  
Protein Binding Assay ◽  
Induced Changes ◽  
Camp Levels ◽  
Human Cervix

Abstract. The effects of porcine relaxin on the levels of cAMP in human cervical tissue were studied in vitro. The specimens were obtained by needle biopsy from women undergoing hysterectomy, legal abortion in the first trimester or elective Casearean section at term, and were incubated in Krebs-Ringer buffer for 15 min in the presence of porcine relaxin (5 μg/ml, 3000 GPU/mg). cAMP was determined using a modified protein binding assay. The concentration of cAMP was higher in pregnant than in non-pregnant women. Relaxin stimulated the production of cAMP in the 7th–8th week of gestation and at term but did not significantly alter the cervical cAMP levels in neither non-pregnant women nor in women in the 10th–12th week of pregnancy. Previous studies have shown that porcine relaxin reduces collagen synthesis in tissue from the human cervix and lower uterine segment. The present observations indicate that these effects can be mediated by cAMP.

Adenosine 3',5'-monophosphate in relation to inhibition of cervical smooth muscle activity in early pregnant women

1991 ◽  
Vol 125 (2) ◽  
pp. 122-126 ◽  
Author(s):  
A. Norström ◽  
I. Bryman
Keyword(s):  
Smooth Muscle ◽  
Pregnant Women ◽  
Muscle Activity ◽  
Needle Biopsy ◽  
Contractile Activity ◽  
First Trimester ◽  
Prostaglandin E ◽  
Cervical Tissue ◽  
First Trimester Of Pregnancy ◽  
Camp Levels

Abstract. Contractile activity was registered in strips of cervical tissue obtained by needle biopsy from women in the first trimester of pregnancy. Dibutyryl cyclic adenosine-3',5'-monophosphate (5 × 10−6mol/l), isobutyryl methylxanthine (10−4mol/l), and forskolin (10−510−4mol/l), the latter two drugs known to increase the levels of endogenous cAMP, inhibited spontaneous muscle activity. The levels of tissue cAMP were determined in strips during relaxation induced by prostaglandin E2 or purified porcine relaxin and compared with cAMP levels in strips from the same women during contractile activity. Exposure to prostaglandin E2 but not to relaxin was followed by increased levels of cAMP. It is suggested that cAMP has a role as a second messenger in the prostaglandin E2-mediated relaxation of cervical smooth muscle.

Alpha 1-adrenoceptor-mediated phosphoinositide breakdown and inotropic responses in diabetic hearts

1991 ◽  
Vol 260 (2) ◽  
pp. H557-H562 ◽  
Author(s):  
H. Xiang ◽  
J. H. McNeill
Keyword(s):  
Positive Inotropic Effect ◽  
Binding Assay ◽  
Diabetic Rats ◽  
Diabetic Rat ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Protein Binding Assay ◽  
Rat Hearts ◽  
Induced Changes ◽  
Inotropic Responses

The mechanism(s) involved in diabetes-induced changes in the heart is still unclear, but one defect appears to occur in the alpha 1-adrenoceptor system. We evaluated the possibility that the changes in the inotropic responsiveness to alpha 1-adrenoceptor stimulation in streptozotocin-diabetic rat hearts may be linked to altered phosphoinositide turnover. Stimulation of alpha 1-adrenoceptor by norepinephrine (in the presence of propranolol) in right ventricles resulted in the formation of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] [measured with an Ins(1,4,5)P3 protein binding assay kit] in a time- and concentration-dependent manner in both control and diabetic rats. The increase in Ins(1,4,5)P3 preceded the increase in the norepinephrine-mediated positive inotropic effect. Diabetic hearts showed a greater maximum inotropic response to norepinephrine stimulation and also had higher Ins(1,4,5)P3 levels. These observations suggest that the changes in Ins(1,4,5)P3 levels may be implicated in the increased inotropic responsiveness to alpha 1-adrenoceptor stimulation in diabetic hearts. Ca2+ overload, induced by Ins(1,4,5)P3, could further be involved in the development of diabetic cardiomyopathy.

In vitro RNA-protein Binding Assay by UV Crosslinking

BIO-PROTOCOL ◽  
2012 ◽  
Vol 2 (21) ◽  
Author(s):  
Hailong Zhang ◽  
Muxiang Zhou
Keyword(s):  
Protein Binding ◽  
Binding Assay ◽  
Uv Crosslinking ◽  
Protein Binding Assay

SERUM THYROXINE-BINDING GLOBULIN: DETERMINATION BY COMPETITIVE LIGAND-BINDING ASSAY IN THYROID DISEASE AND PREGNANCY

1977 ◽  
Vol 85 (2) ◽  
pp. 314-324 ◽  
Author(s):  
C. Mulaisho ◽  
R. D. Utiger
Keyword(s):  
Ligand Binding ◽  
Pregnant Women ◽  
Binding Assay ◽  
Normal Subjects ◽  
First Trimester ◽  
Ligand Binding Assay ◽  
Hormone Production ◽  
Thyroxine Binding Globulin ◽  
Serum Thyroxine ◽  
Competitive Ligand

ABSTRACT Serum thyroxine-binding globulin (TBG) concentrations were measured by competitive ligand-binding assay in normal subjects and patients with a variety of abnormalities of thyroid hormone production or TBG production. The mean serum TBG concentration in 73 normal adults was 3.4 ± 0.6 mg/100 ml. No correlations with age, sex or serum thyroxine (T4) or triiodothyronine (T3) concentrations were found. Serum TBG concentrations were normal in patients with hyperthyroidism. They were significantly elevated in hypothyroid patients, and fell to normal with thyroxine treatment. In pregnant women, serum TBG concentrations were markedly increased, being 7.1 ± 1.2 (mean ± sd) mg/100 ml in women in their first trimester, 9.0 ± 1.0 mg/100 ml in the second trimester and 8.9 ± 1.6 mg/100 ml in the third trimester. There was a positive correlation between serum TBG concentration and both serum T4 and serum T3 concentrations in pregnant women. Competitive ligand-binding assay is a simple and reliable method for TBG assay and yields results similar to those of electrophoretic saturation techniques.

CORTICOSTEROID RELEASE BY ADRENAL TISSUE FROM FOETAL AND NEWBORN LAMBS IN RESPONSE TO CORTICOTROPHIN STIMULATION IN A PERIFUSION SYSTEM IN VITRO

1973 ◽  
Vol 58 (1) ◽  
pp. 75-87 ◽  
Author(s):  
DENISE MADILL ◽  
J. M. BASSETT
Keyword(s):  
Adrenal Glands ◽  
Binding Assay ◽  
Developmental Changes ◽  
Acth Stimulation ◽  
Adult Sheep ◽  
Adrenal Tissue ◽  
A Value ◽  
Protein Binding Assay ◽  
Perifusion System

SUMMARY Adrenal glands from 24 foetal lambs (100–145 days gestation) and eight postnatal lambs (birth to 25 days of age) were used to study developmental changes in the responsiveness of the lamb adrenal to stimulation by synthetic adrenocorticotrophin (ACTH). A continuous flow incubation system in vitro (perifusion) was used in these studies. Addition of ACTH to the perifusion medium (0·3 μg Synacthen/ml) significantly increased the rate of corticosteroid release (as measured by a protein-binding assay) by adrenal tissue from foetuses of all ages studied. With tissue from foetuses of 137 days gestation or less, corticosteroid release during ACTH stimulation was < 2 μg/100 mg tissue in the 3 h following addition of ACTH to the perifusion medium. With tissue from foetuses older than this, the rate of corticosteroid release stimulated by ACTH increased as foetal age increased. At 145 days gestation (just before birth) release was 6·8 μg/100 mg tissue and in postnatal lambs less than 1 day old it was 12·3 μg/100 mg tissue during the 3 h following the start of stimulation. This increase in the response of the foetal adrenal to ACTH stimulation paralleled the increases in adrenal weight and plasma corticosteroid concentration which occurred before birth. With tissue from older lambs ACTH-stimulated corticosteroid release was 5·5 μg/100 mg tissue/3 h, a value comparable to that of 5·6 μg/100 mg tissue/3 h obtained with adrenal tissue from adult sheep. Separation of corticosteroids in the perifusion medium by chromatography on LH 20 Sephadex indicated that adrenals from younger foetuses (< 137 days gestation) released approximately equal amounts of cortisol and corticosterone, whereas adrenals from older foetuses and postnatal lambs released mainly cortisol. Pituitary tissue from all foetuses stimulated corticosteroid release from adrenal tissue of adult sheep, implying that foetal pituitary stores of ACTH do not limit development of foetal adrenal cortex function during the last third of gestation. The results indicate that there is a marked increase during the last week of gestation in the responsiveness of the foetal adrenal to ACTH stimulation and that this increase is related to maturation of corticosteroid biosynthetic pathways during this period.

Determination of erythrocyte folate by competitive protein binding assay preceded by extraction.

1976 ◽  
Vol 22 (7) ◽  
pp. 982-992 ◽  
Author(s):  
E Mortensen
Keyword(s):  
Protein Binding ◽  
Binding Assay ◽  
Extraction Procedure ◽  
Molecular Configuration ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
Erythrocyte Folate

Abstract Determination of the concentration of erythrocyte folate by means of competitive protein binding assay critically depends on the extraction procedure applied. Results will be influenced by variable factors such as the in vitro age of the blood samples, the degree of hemolysis, the presence of ascorbic acid, and the pH during extraction and elimination of proteins. The radioassay is strongly influenced by the pH of the final reaction mixture, the method used to separate free and protein-bound molecules, and the molecular configuration of the folates present. Based on experimental results presented, I describe a method for the determination of erythrocyte folate.

Determination of cyclosporine by a competitive binding assay with cyclophilin

1991 ◽  
Vol 37 (3) ◽  
pp. 403-410 ◽  
Author(s):  
G Steiner ◽  
B Küllinger ◽  
G Mayer ◽  
Y Jie ◽  
H Leibl ◽  
...  
Keyword(s):  
Binding Assay ◽  
Biological Fluids ◽  
Bioactive Metabolites ◽  
Binding Affinities ◽  
Competitive Binding Assay ◽  
Protein Binding Assay ◽  
Analytical Recovery ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding

Abstract A competitive protein-binding assay for cyclosporine based on use of the intracellular cyclosporine-binding protein cyclophilin (CYP) was used to measure cyclosporin A (CsA) and its bioactive metabolites in whole blood. CYP from cytoplasmic extracts or erythrocyte lysates was applied in the binding assay with use of [3H]CsA as tracer and charcoal adsorption for separating bound from free tracer. Binding affinities of various CsA analogs and metabolites correlated well with their reported in vitro immunosuppressive activities. The assay detected as little CsA as 50 micrograms/L (1 g = 0.832 mmol of CsA), analytical recovery was greater than 80%, and CVs were less than 8% for intra-assay and less than 11% for interassay precision in the range of 150-1000 micrograms/L. We used this assay to measure CsA concentrations in blood and compared the results with those measured by HPLC or by CsA-specific (monoclonal) and CsA-nonspecific (polyclonal) radioimmunoassays. Binding assay results were, in nearly all cases, less than those measured by the nonspecific RIA and frequently greater than 20% above the values determined by the CsA-specific assays. Individual patients had pronounced differences in the relative proportions of CsA, CYP-binding (bioactive) metabolites, and cross-reacting CsA metabolites. Because the presence of bioactive metabolites may considerably contribute to the immunosuppressive activity of CsA, we consider the binding assay clinically useful for measuring CsA in biological fluids.

CHARACTERISTICS OF THE CYCLIC 3′, 5′-AMP FORMATION IN ISOLATED OVARIAN FOLLICLES FROM PMSG-TREATED IMMATURE RATS AFTER STIMULATION IN VITRO WITH GONADOTROPHINS AND PROSTAGLANDINS

1974 ◽  
Vol 77 (3) ◽  
pp. 559-574 ◽  
Author(s):  
Lars Nilsson ◽  
Sten Rosberg ◽  
Kurt Ahrén
Keyword(s):  
Binding Assay ◽  
Prostaglandin E ◽  
Maximal Effect ◽  
Bicarbonate Buffer ◽  
Protein Binding Assay ◽  
Immature Rats ◽  
Significant Release ◽  
Time Courses ◽  
Ovulatory Follicles

ABSTRACT Isolated pre-ovulatory follicles from PMSG-injected immature rats, described in a previous publication, have been used for the investigation of the pattern of cyclic 3′,5′-AMP synthesis after in vitro addition of gonadotrophins and prostaglandins. The follicles were incubated in a modified Krebs bicarbonate buffer for periods up to 4 h and the cyclic 3′,5′-AMP content in tissue and medium was analyzed with the protein binding assay of Gilman (1970). Addition of LH, FSH or prostaglandin E2 (PGE2) increased the cyclic 3′,5′-AMP level in follicular tissue dramatically. HCG and TSH, but not STH or prolactin, also induced a similar increase. The effect of FSH was blocked by a highly specific antiserum to LH. The effect of LH had a maximum after approximately 1 h of incubation, whereas the maximal effect of PGE2 was seen between 5 and 15 min of incubation. When LH was present in the incubation medium a significant release of cyclic 3′,5′-AMP into the medium occurred. Theophylline potentiated the effects of LH and PGE2. The different time-courses of the effects of LH and PGE2 on the cyclic 3′,5′-AMP production by follicular tissue are discussed in relation to the hypothesis of prostaglandins as obligatory mediators of all LH effects on the ovary (Kuehl et al. 1970).

Systemic Effects of ADP-induced Platelet Aggregation and Their Modification by Aspirin and by Pyridinolcarbamate

1973 ◽  
Vol 30 (01) ◽  
pp. 178-190 ◽  
Author(s):  
Itsuro Kobayashi ◽  
Paul Didisheim
Keyword(s):  
Platelet Aggregation ◽  
Venous Pressure ◽  
Systemic Effects ◽  
Central Venous ◽  
Platelet Aggregates ◽  
Induced Changes ◽  
Second Wave ◽  
Carotid Arterial ◽  
Arterial Po2

SummaryADP, AMP, or ATP was injected rapidly intravenously in rats. ADP injection resulted in the f olio wing transient changes: a drop in platelet count, a rise in central venous pressure, a fall in carotid arterial PO2, bradycardia, arrhythmia, flutter-fibrillation, and arterial hypotension. AMP and ATP produced some of these same effects; but except for hypotension, their frequency and severity Avere much less than those following ADP.Prior intravenous administration of acetylsalicylic acid or pyridinolcarbamate, two inhibitors of the second wave of ADP-induced platelet aggregation in vitro, significantly reduced the frequency and severity of all the above ADP-induced changes except hypotension. These observations suggest that many of the changes (except hypotension) observed to follow ADP injection are produced by platelet aggregates which lodge transiently in various microcirculatory beds then rapidly disaggregate and recirculate.

Optimisation of tactics of conducting pregnancy and labours at women after auxiliary reproductive technologies

2016 ◽  
pp. 160-164
Author(s):  
D.N. Maslo ◽  
Keyword(s):  
Pregnant Women ◽  
Resistance Index ◽  
First Trimester ◽  
Reproductive Technologies ◽  
Premature Delivery ◽  
Control Group ◽  
Uterine Arteries ◽  
Placental Dysfunction ◽  
Placental Blood ◽  
Two Stages

The objective: frequency decrease perinatal pathologies at women after ART on the basis of studying clinical-ehografical, endocrinological, biochemical, dopplerometrical, cardiotokografical and morphological researches, and also improvement of algorithm of diagnostic and treatment-and-prophylactic actions. Patients and methods. The work basis is made spent by us from 2012 on 2015 by complex inspection of 300 pregnant women from which 250 were after ART and 50 – firstlabours which pragnency without ART, and also their newborns. For the decision of an object in view of research spent to two stages. At 1 stage spent prosperctive research which included 150 pregnant women: з them 100 women pregnancy at which has come out ART (1 group) and 50 healthy women (control group). At 2 stage spent prospective randomization in which result of patients after ART have divided on two equal groups by therapy principle: 2 basic group - 75 pregnant women after ART at which used the algorithm improved by us; 3 group of comparison - 75 pregnant women after ART which have been spent on the standard treatment-and-prophylactic actions. Results. The results suggest that women after using ART is a high frequency of reproductive losses in the first trimester (10.0%), 3.0% of spontaneous abortion from 16 to 22 weeks, and 3.0% "early" premature delivery (22 to 28 weeks of pregnancy). The frequency of violations of the functional state of placenta in women after using IVF is 63.0%, which is the main cause of high levels of perinatal losses (40.0 ‰), and delivery by cesarean section (96.0%). Placental dysfunction in women after using ART characterized by retrohorialnyh hematoma (21.0%); size mismatch fruit (30.0%) and hypertonicity of the uterus (73.0%) against changes in fruit-placental blood flow - increased resistance index in umbilical artery and increased vascular resistance in the uterine arteries. Endocrinological and biochemical changes in placental dysfunction in women after using IVF starting from 28 weeks of pregnancy and are in significant reduction in progesterone, placental b1-microglobulin, B2-microglobulin of fertility and trophic в-glycoprotein. Conclusion. The received results: use of the algorithm of diagnostic and treatment-and-prophylactic actions improved by us allows to lower frequency of spontaneous interruption of pregnancy till 22 weeks – from 13.0% to 5.7%; «early» premature birth – from 3.0% to 1.0%; placentary dysfunction from 63.0% to 40.6%; cesarean sections – from 96.0% to 56.5%, and also perinatal losses – from 40.0‰ to 16.2‰. Key words: pregnancy, childbirth, auxiliary reproductive technologies.

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