PLASMA CORTICOSTEROIDS AND THE EFFECT OF ADRENOCORTICOTROPHIN IN A MACROPODID MARSUPIAL (SETONIX BRACHYURUS, QUOY & GAIMARD)

1977 ◽  
Vol 75 (3) ◽  
pp. 409-418 ◽  
Author(s):  
I. R. McDONALD ◽  
S. D. BRADSHAW
Keyword(s):  
Plasma Proteins ◽  
Binding Capacity ◽  
Control Value ◽  
High Affinity ◽  
Peripheral Plasma ◽  
Constant Rate ◽  
Moderate Disturbance ◽  
Maximum Binding Capacity ◽  
Repeated Sampling ◽  
The Mean

The total corticosteroid concentrations in the peripheral plasma of unanaesthetized, undisturbed quokkas were 0·75 ± 0·10 (s.e.m.) and 0·93 ± 0·14 μg/100 ml in male and female quokkas respectively. Repeated sampling for periods of 36–49 h disclosed irregular fluctuations over the range 0·4–5·0 μg/100 ml, but no evidence for a regular periodicity. The major corticosteroid was usually cortisol but corticosterone contributed 25–50% of the total unstimulated corticosteroid concentration. Relatively minor concentrations of 11-deoxycortisol were detected. Constant-rate i.v. infusion of ACTH caused a significant increase in the concentration of total corticosteroids in the plasma; this increase was detectable at a dose of 0·05 i.u. ACTH/kg/h, and rose to approximately 15 times the control value at a dose of 2·0 i.u./kg/h. This increase was due mainly to a change in the concentration of cortisol. Synthetic (β1–24) and porcine ACTH were equipotent. The sensitivity of the quokka to ACTH was approximately one-tenth that of another marsupial (Trichosurus vulpecula) or 1/160 that of man. Moderate disturbance increased the concentration of corticosteroids in the plasma to the same level as that caused by infusion of 0·1 i.u. ACTH/kg/h, the increase being mainly in the cortisol fraction. High-affinity binding of cortisol and corticosterone by plasma proteins was demonstrated. The maximum binding capacities for cortisol were 3·89 ± 0·5 and 3·02 ± 0·6 μg/100 ml in female and male quokkas respectively. The mean association constant was 3·2 × 1081/mol at 4 °C and 5·5 × 1071/mol at 36 °C. The maximum binding capacity for corticosterone was approximately one-third that of cortisol.

scholarly journals Presence of D4 dopamine receptors in human prefrontal cortex: a postmortem study

2007 ◽  
Vol 29 (2) ◽  
pp. 148-152 ◽  
Author(s):  
Donatella Marazziti ◽  
Antonio Lucacchini ◽  
Stefano Baroni ◽  
Laura Betti ◽  
Mario Catena ◽  
...  
Keyword(s):  
Prefrontal Cortex ◽  
Dopamine Receptors ◽  
Binding Capacity ◽  
Assay Method ◽  
Nonspecific Binding ◽  
D3 Receptors ◽  
High Affinity ◽  
Maximum Binding Capacity ◽  
Tissue Homogenates ◽  
The Mean

OBJECTIVE: The aim of our study was to explore the presence and the distribution of D4 dopamine receptors in postmortem human prefrontal cortex, by means of the binding of [³H]YM-09151-2, an antagonist that has equal affinity for D2, D3 and D4 receptors. It was therefore necessary to devise a unique assay method in order to distinguish and detect the D4 component. METHOD: Frontal cortex samples were harvested postmortem, during autopsy sessions, from 5 subjects. In the first assay, tissue homogenates were incubated with increasing concentrations of [³H]YM-09151-2, whereas L-745870, which has a high affinity for D4 and a low affinity for D2/D3 receptors, was used as the displacer. In the second assay, raclopride, which has a high affinity for D2/D3 receptors and a low affinity for D4 receptors, was used to block D2/D3. The L-745870 (500 nM) was added to both assays in order to determine the nonspecific binding. RESULTS: Our experiments revealed the presence of specific and saturable binding of [³H]YM-09151-2. The blockade of D2 and D3 receptors with raclopride ensured that the D4 receptors were labeled. The mean maximum binding capacity was 88 ± 25 fmol/mg protein, and the dissociation constant was 0.8 ± 0.4 nM. DISCUSSION AND CONCLUSIONS: Our findings, although not conclusive, suggest that the density of D4 receptors is low in the human prefrontal cortex.

Specific binding of serotonin in rat lung

1985 ◽  
Vol 248 (1) ◽  
pp. E58-E63 ◽  
Author(s):  
D. K. Das ◽  
H. Steinberg
Keyword(s):  
Binding Capacity ◽  
Specific Binding ◽  
Direct Action ◽  
High Capacity ◽  
Mitochondrial Fraction ◽  
Temperature Sensitive ◽  
Inner Mitochondrial Membrane ◽  
High Affinity ◽  
Single Temperature ◽  
Maximum Binding Capacity

Mammalian lungs have been shown to store and to inactivate serotonin (5-HT) by an active process involving uptake and metabolism. 5-HT has direct action on lung including constrictor effects of pulmonary vascular and tracheobronchial smooth muscle, suggesting the presence of 5-HT receptors in lung. We have identified specific 5-HT binding of high affinity to the different lung portions and have shown that there was a different capacity for this binding. Two different 5-HT-binding capacities are present in a purified mitochondrial fraction. Saturation analysis of 5-[3H]HT binding to outer mitochondrial membranes demonstrates a single, temperature-sensitive, high-affinity and high-capacity binding (Kd = 8.3 +/- 1.2 nM, maximum binding capacity = 0.819 +/- 0.046 pmol/mg protein). The dissociation constant of inner mitochondrial membrane demonstrates a low-capacity site (Kd = 25.2 +/- 2.2 nM, maximum binding capacity = 0.453 +/- 0.037 pmol/mg protein). The purified microsomal fraction of lung exhibits a high-capacity binding site for 5-[3H]HT (Kd = 14.8 +/- 1.6 nM, maximum binding capacity = 0.760 +/- 0.03 pmol/mg protein). In addition to the lung being the major site for its inactivation, the presence of several specific 5-HT receptors may be related to some of the known 5-HT actions in lung and may suggest other unknown actions of this amine.

ATP-MgCl2 treatment after trauma-hemorrhage/resuscitation increases hepatocyte P2-purinoceptor binding capacity

1994 ◽  
Vol 266 (6) ◽  
pp. R1810-R1815
Author(s):  
M. S. Mahmoud ◽  
P. Wang ◽  
S. R. Hootman ◽  
S. S. Reich ◽  
I. H. Chaudry
Keyword(s):  
Binding Capacity ◽  
Scatchard Analysis ◽  
Binding Characteristics ◽  
Beneficial Effects ◽  
High Affinity ◽  
Receptor Component ◽  
Receptor Dynamics ◽  
Maximum Binding Capacity ◽  
High Affinity Receptor ◽  
Affinity Receptor

Although our studies indicate that P2-purinoceptor binding capacity decreases after hemorrhage and resuscitation, it is not known whether ATP-MgCl2 administration after hemorrhage has any beneficial effects on the receptor dynamics. To study this, we performed laparotomy (i.e., trauma induced) on rats and bled them to and maintained them at a mean arterial pressure of 40 mmHg until 40% of maximum bleedout volume was returned in the form of Ringer lactate (RL). The animals were then resuscitated with 3 times the volume of maximum bleedout with RL over 45 min followed by 2 times RL along with ATP-MgCl2 (50 mumol/kg body wt) over 95 min. Hepatocytes were isolated at 4, 17, and 27 h after resuscitation. P2-purinoceptor binding characteristics were determined by using [alpha-35S]ATP. Scatchard analysis revealed high-affinity and low-affinity receptor components in the hepatocytes isolated from sham-operated or hemorrhaged animals with or without ATP-MgCl2 infusion. ATP-MgCl2 ameliorated and subsequently restored the decreased maximum binding capacity (Bmax) of the high-affinity receptor component and significantly improved Bmax of the low-affinity receptor component. ATP-MgCl2 administration also produced a progressive enhancement in the affinity of the low-affinity receptor component. Thus the beneficial effects of ATP-MgCl2 observed after trauma-hemorrhage and resuscitation may be, in part, due to the restoration of P2-purinoceptor binding capacity and the enhancement of the receptor affinity.

Uptake of 3,5,3′-l-tri-iodothyronine in human erythrocytes

1989 ◽  
Vol 121 (3) ◽  
pp. 585-591 ◽  
Author(s):  
K. Yamauchi ◽  
R. Horiuchi ◽  
H. Takikawa
Keyword(s):  
Binding Sites ◽  
Binding Capacity ◽  
Human Erythrocytes ◽  
The Other ◽  
Transport Pathway ◽  
High Affinity ◽  
High Affinity Site ◽  
Maximum Binding Capacity ◽  
Energy Dependent ◽  
Dependent Pathway

ABSTRACT The mechanisms of 3,5,3′-l-tri-iodothyronine (T3) uptake into human erythrocytes were examined. Purified membranes of human erythrocytes were shown to have two classes of T3-binding sites with one being a high-affinity site (dissociation constant, 59·2±17·8 nmol/l; maximum binding capacity, 344·3 ± 95·5 fmol/μg protein). Furthermore, it was shown that there were two pathways for T3 uptake in human erythrocytes; one was saturable, stereospecific (T3»thyroxine > 3,5,3′-d-tri-iodothyronine), energydependent and dominant at 15 °C; the other was not displaced by unlabelled T3 and was energyindependent but did not occur by passive diffusion. The former pathway which, it is suggested, is a receptor-mediated transport pathway, was inhibited by monodansylcadaverine, phloretin or oligomycin at 15 or 37 °C, but the latter pathway was not inhibited by these inhibitors. Our results strongly suggest that uptake of T3 by the energy-independent pathway became predominant over the energy-dependent pathway at 37 °C and accounted for 83% of total T3 uptake of human erythrocytes. Journal of Endocrinology (1989) 121, 585–591

Annual and diurnal patterns of plasma calcium fractions in anuran Rana pipiens

1985 ◽  
Vol 249 (3) ◽  
pp. R290-R295
Author(s):  
D. R. Robertson
Keyword(s):  
Plasma Proteins ◽  
Rana Pipiens ◽  
Binding Capacity ◽  
Apparent Movement ◽  
Total Calcium ◽  
Annual Cycles ◽  
Diurnal Patterns ◽  
Calcium Complexes ◽  
The Mean ◽  
C 30

Ionized calcium [Ca2+] in the amphibian Rana pipiens was measured over 18 mo. An annual variation about the mean of 1.10 mmol/l was observed that peaked in April (1.36 mmol/l) and attained minimal levels in October (0.84 mmol/l). Plasma total calcium [Ca]t, plasma proteins, and the bound calcium fraction also exhibited annual oscillations. The annual mean binding capacity of plasma proteins for calcium was established at 0.0175 mmol/g. Of the plasma total calcium in spring animals, nonprotein-bound calcium complexes [Ca]c were less than 3% of the total whereas 80% was composed of [Ca2+] and 17% as protein-bound calcium [Ca]pb. In autumn [Ca2+] composed 40%, [Ca]pb 30%, and [Ca]c 30% of plasma total calcium. An equilibrium exists between [Ca2+] and [Ca]c, which maintains a combined annual mean of 1.48 mmol/l calcium. Diurnal variations superimposed on the annual cycles are modulated by apparent movement of plasma water and an intrinsic 12-h rhythm for [Ca2+]. No definite associations could be made with the entrainment of specific rhythms of the three calcium fractions with the light-dark cycle.

EFFECTS OF ACUTE NEUROTROPHIC STRESS ON PERIPHERAL METABOLISM OF CORTISOL IN CONSCIOUS MALE GUINEA-PIGS

1981 ◽  
Vol 89 (3) ◽  
pp. 443-449 ◽  
Author(s):  
M. MANIN ◽  
P. DELOST
Keyword(s):  
Adult Male ◽  
Guinea Pigs ◽  
Binding Capacity ◽  
Apparent Volume ◽  
Volume Of Distribution ◽  
Metabolic Clearance ◽  
Control Value ◽  
Cortisol Metabolism ◽  
Free Cortisol ◽  
The Mean

Cortisol metabolism was studied in conscious adult male guinea-pigs subjected to a neurotrophic stress (immobilization and stimulation by light for 3 h). The disappearance curves of tracer quantities of [3H]cortisol were represented by a two-pool model. In stressed animals, there was a marked increase in the mean plasma level of cortisol (184% of control value; P <0·001) and in the metabolic clearance rate (MCR; 17% of control value; 0·001 <P <0·01). This rise in the MCR of plasma cortisol resulted from an increase in the mean total apparent volume of distribution (49%, P < 0·001). The lack of significant differences in the slopes of the second exponential phase of the disappearance curves indicated that the stress did not significantly increase the half-life of cortisol. The mean binding capacity of transcortin for cortisol (ST) was significantly higher in the animals which had been subjected to the neurotrophic stress than in the control guinea-pigs (0·02 < P <0·05). However, ST values remained very low and accounted for the very high levels of free cortisol found after the stress. The results suggest that the raised concentrations of unbound cortisol found in the plasma of conscious adult male guinea-pigs in response to neurotrophic stress reflect a hypersecretion of corticosteroid.

THE BINDING OF CORTISOL BY OVINE PLASMA PROTEINS

1967 ◽  
Vol 37 (3) ◽  
pp. 261-268 ◽  
Author(s):  
J. Y. F. PATERSON ◽  
F. HILLS
Keyword(s):  
Human Plasma ◽  
Plasma Proteins ◽  
Binding Capacity ◽  
Equilibrium Dialysis ◽  
Affinity Constant ◽  
Albumin Solution ◽  
Plasma Albumin ◽  
The Mean ◽  
Mean Concentration

SUMMARY Albumin was isolated from ovine plasma and its affinity for cortisol was determined by equilibrium dialysis at 37°. The value of Ka[σpa] for a 1 % (w/v) albumin solution was 0·275 which is similar to the value for human plasma albumin. The affinity constant of transcortin in ovine plasma was determined by equilibrium dialysis of diluted plasma at several concentrations of cortisol. The value found, Kt (37°) = 0·87 x 108 l./mole, is close to that found for human plasma transcortin by Mills (1962). The concentration of transcortin in ovine plasma, expressed as cortisolbinding capacity, was 6–49 μg. (mean 24 μg.) cortisol/l. These concentrations are much lower than those found in human plasma. The observation of Lindner (1964) that cortisol binding capacity did not increase during pregnancy in sheep has been confirmed. In sheep which were accustomed to handling, the mean concentration of cortisol in plasma was 17·8 μg./l. and of this amount 59% was bound to transcortin, 19 % to albumin and 22 % was not bound to protein.

PROTEIN BINDING OF PLASMA CORTISOL IN THE FOETAL LAMB NEAR TERM

1975 ◽  
Vol 67 (3) ◽  
pp. 333-341 ◽  
Author(s):  
R. J. FAIRCLOUGH ◽  
G. C. LIGGINS
Keyword(s):  
Plasma Proteins ◽  
Binding Capacity ◽  
Equilibrium Dialysis ◽  
Progressive Increase ◽  
Mean Value ◽  
Affinity Constant ◽  
Total Cortisol ◽  
The Mean ◽  
Near Term ◽  
Foetal Lamb

SUMMARY Binding of cortisol to plasma proteins was studied in the foetal lamb by equilibrium dialysis at 37 °C. At 122 days of pregnancy the mean level of transcortin expressed as cortisolbinding capacity was 28 ± 6 (s.d.) ng cortisol/ml plasma. During the last 14 days of pregnancy there was a progressive increase in transcortin-binding capacity to 85 ± 14 ng cortisol/ ml plasma. A sharp increase in the concentration of both protein-bound and unbound cortisol was observed over the same period. A rise in the concentration of total cortisol from around 3 to 42 ng/ml was associated with an increase in unbound cortisol from 0·2 to a maximum of 2·1 ng/ml. The concentration of albumin-bound cortisol was approximately equal to that of unbound cortisol. The mean value for the transcortin–cortisol affinity constant was 1·15 × 108 l/mol. It is concluded that an increase in transcortin-binding capacity is partly responsible for the prepartum increase of corticosteroid levels observed in normal foetal lambs.

Investigation of the Free and Total Thyroxine-binding Capacity of Plasma Proteins in Thyroid Disorders

1971 ◽  
Vol 10 (04) ◽  
pp. 299-304
Author(s):  
József Takó ◽  
János Fischer ◽  
Jusztina Juhász ◽  
Ilona Sztraka ◽  
István Kapus ◽  
...  
Keyword(s):  
Blood Cell ◽  
Thyroid Function ◽  
Oral Contraceptives ◽  
Red Blood Cell ◽  
Plasma Proteins ◽  
Binding Capacity ◽  
Thyroid Disorders ◽  
Thyroid Function Tests ◽  
Total Thyroxine

SummaryThe results of thyroid function tests have been compared with data on the thyroxine-binding capacity of plasma proteins in hyper-, hypo- and euthyroid cases, the latter including women taking oral contraceptives (Infecundin). It was found that there exists a significant correlation of exponential nature between the in vitro red blood cell 125I-triiodothyronine uptake (RCU) and the free thyroxine-binding capacity of the thyroxine-inding globulin (TBG).

RADIOIMMUNOASSAY OF OESTRONE AND OESTRADIOL IN THE PERIPHERAL PLASMA OF THE DOMESTIC FOWL IN VARIOUS PHYSIOLOGICAL STATES AND OF HYPOPHYSECTOMIZED AND OVARIECTOMIZED FOWLS

1974 ◽  
Vol 75 (1) ◽  
pp. 133-140 ◽  
Author(s):  
B. E. Senior
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Diethyl Ether ◽  
Bovine Serum ◽  
Domestic Fowl ◽  
Laying Hens ◽  
Peripheral Plasma ◽  
The Mean ◽  
Precision And Accuracy ◽  
Chicken Ovary

ABSTRACT A radioimmunoassay was developed to measure the levels of oestrone and oestradiol in 0.5–1.0 ml of domestic fowl peripheral plasma. The oestrogens were extracted with diethyl ether, chromatographed on columns of Sephadex LH-20 and assayed with an antiserum prepared against oestradiol-17β-succinyl-bovine serum albumin using a 17 h incubation at 4°C. The specificity, sensitivity, precision and accuracy of the assays were satisfactory. Oestrogen concentrations were determined in the plasma of birds in various reproductive states. In laying hens the ranges of oestrone and oestradiol were 12–190 pg/ml and 29–327 pg/ml respectively. Levels in immature birds, in adult cockerels and in an ovariectomized hen were barely detectable. The mean concentrations of oestrone and oestradiol in the plasma of four non-laying hens (55 pg/ml and 72 pg/ml respectively) and one partially ovariectomized hen (71 pg/ml and 134 pg/ml respectively) were well within the range for laying hens. It is evident that the large, yolk-filled follicles are not the only source of oestrogens in the chicken ovary.

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