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.

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.

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

Demonstration of specific dopamine receptors on human pituitary adenomas

1987 ◽  
Vol 114 (4) ◽  
pp. 595-602 ◽  
Author(s):  
Masafumi Koga ◽  
Haruyoshi Nakao ◽  
Masayo Arao ◽  
Bunzo Sato ◽  
Keizo Noma ◽  
...  
Keyword(s):  
Dopamine Receptors ◽  
Binding Sites ◽  
Pituitary Adenomas ◽  
Binding Capacity ◽  
Affinity Binding ◽  
Human Pituitary ◽  
Human Pituitary Adenoma ◽  
High Affinity ◽  
Number Of Binding Sites ◽  
Dissociation Constant Kd

Abstract. Dopamine receptors on human pituitary adenoma membranes were characterized using [3H] spiperone as the radioligand. The specific [3H]spiperone binding sites on prolactin (PRL)-secreting adenoma membranes were recognized as a dopamine receptor, based upon the data showing high affinity binding, saturability, specificity, temperature dependence, and reversibility. All of 14 PRL-secreting adenomas had high affinity dopamine receptors, with a dissociation constant (Kd) of 0.85 ±0.11 nmol/l (mean ± sem) and a maximal binding capacity (Bmax) of 428 ± 48.6 fmol/mg protein. Among 14 growth hormone (GH)-secreting adenomas examined, 8 (57%) had dopamine receptors with a Kd of 1.90 ± 0.47 nmol/l and a Bmax of 131 ± 36.9 fmol/mg protein. Furthermore, 15 of 24 (58%) nonsecreting pituitary adenomas also had dopamine receptors with a Kd of 1.86 ± 0.37 nmol/l and a Bmax of 162 ± 26.0 fmol/mg protein. These results indicate that some GH-secreting adenomas as well as some nonsecreting pituitary adenomas contain dopamine receptors. But their affinity and number of binding sites are significantly lower (P < 0.05) and fewer (P < 0.001) respectively, than those in PRL-secreting adenomas.

THE EFFECT OF ALTERATION OF THYROXINE BINDING CAPACITY ON THE DIALYZABLE AND ABSOLUTE FRACTIONS OF TRIIODOTHYRONINE IN CIRCULATION

1973 ◽  
Vol 72 (2) ◽  
pp. 265-271 ◽  
Author(s):  
J. H. Dussault ◽  
D. A. Fisher ◽  
J. T. Nicoloff ◽  
V. V. Row ◽  
R. Volpe
Keyword(s):  
Correlation Coefficient ◽  
Inverse Relationship ◽  
Binding Capacity ◽  
Hormone Concentration ◽  
Thyroxine Binding Globulin ◽  
Serum Thyroxine ◽  
A Value ◽  
The Mean ◽  
Male Subjects ◽  
Female Subjects

ABSTRACT In order to determine the effect of alterations in binding capacity of thyroxine binding globulin (TBG) on triiodothyronine (T3) metabolism, studies were conducted in 10 patients with idiopathically low (7 subjects) or elevated (3 subjects) TBG levels and 10 subjects given norethandrolone (7 male subjects) or oestrogen (3 female subjects). Measurements of serum thyroxine (T4) concentration, maximal T4 binding capacity, serum T3 concentration and per cent dialyzable T3 were conducted. Serum T3 was measured both by chemical and radioimmunoassay methods. In patients with idiopathically low TBG, the mean serum T4 concentration was low (2.4 μg/100 ml), the mean serum T3 level low (55 ng/100 ml), the mean per cent dialyzable T3 increased (0.52%), and the calculated free T3 concentration normal (186 pg/100 ml). In patients with idiopathically high TBG levels the mean T4 concentration was high (10.3 μg/100 ml), the mean T3 level slightly elevated (127 ng/100 ml), the% dialyzable T3 low (0.10%) and the calculated free T3 concentration low normal (123 pg/100 ml). The correlation coefficient between the per cent dialyzable T3 and maximal TBG binding capacity in the 20 subjects was 0.68, a value significant at the P < 0.01 level. Thus, alterations in binding capacity of TBG seem to influence T3 and T4 metabolism similarly; the inverse relationship between the % of dialyzable hormone and total hormone concentration tends to keep the absolue levels of free hormones stable.

METHODOLOGICAL STUDY OF THE RADIOIMMUNOASSAY OF HUMAN THYROTROPHIN

1972 ◽  
Vol 71 (1) ◽  
pp. 24-36 ◽  
Author(s):  
Ariel Gordin ◽  
Pirkko Saarinen
Keyword(s):  
Healthy Subjects ◽  
Binding Capacity ◽  
Paper Electrophoresis ◽  
Storage Conditions ◽  
Methodological Study ◽  
Double Antibody ◽  
The Mean ◽  
Specific Activities ◽  
Initial Binding ◽  
In Pregnancy

ABSTRACT An account is given of a methodological study of the double-antibody radioimmunoassay of human TSH, using highly purified labelled human TSH as tracer. It was shown that conventional paper electrophoresis was not adequate for studying the purity of labelled human TSH. When polyvinylchloride (Pevikon®) electrophoresis was used, four subfractions could still be separated, even though, on paper electrophoresis, the material seemed to be homogeneous. Only two of the four Pevikon fractions were immunoreactive. Purification of labelled human TSH by Pevikon electrophoresis also improved the sensitivity of the assay. Specific activities of about 100 mCi/mg gave the highest initial binding capacity, produced least damage to the labelled hormone and showed the best stability of the tracer without influencing the sensitivity of the method. In different storage conditions, labelled human TSH was found to be most stable at −20°C and diluted 1/100. Only in pregnancy did the addition of HCG seem necessary. The mean TSH value in healthy subjects was 3.6 ± 1.4 μU/ml (mean±sd) with a range from 1.6 μU/ml to 8.8 μU/ml.

The Expression of Dopamine Receptors Gene and their Potential Role in Targeting Breast Cancer Cells with Selective Agonist and Antagonist Drugs. Could it be the Novel Insight to Therapy?

2019 ◽  
Vol 16 (2) ◽  
pp. 184-197 ◽  
Author(s):  
Hossein Bakhtou ◽  
Asiie Olfatbakhsh ◽  
Abdolkhaegh Deezagi ◽  
Ghasem Ahangari
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cells ◽  
Dopamine Receptors ◽  
Breast Cancer Cells ◽  
D2 Receptors ◽  
Healthy Individuals ◽  
Agonist And Antagonist ◽  
The Mean ◽  
Mcf 7

Background:Breast cancer is one of the common causes of mortality for women in Iran and other parts of the world. The substantial increasing rate of breast cancer in both developed and developing countries warns the scientists to provide more preventive steps and therapeutic measures. This study is conducted to investigate the impact of neurotransmitters (e.g., Dopamine) through their receptors and the importance of cancers via damaging immune system. It also evaluates dopamine receptors gene expression in the women with breast cancer at stages II or III and dopamine receptor D2 (DRD2) related agonist and antagonist drug effects on human breast cancer cells, including MCF-7 and SKBR-3.Methods:The patients were categorized into two groups: 30 native patients who were diagnosed with breast cancer at stages II and III, with the mean age of 44.6 years and they were reported to have the experience of a chronic stress or unpleasant life event. The second group included 30 individuals with the mean age of 39 years as the control group. In order to determine the RNA concentration in all samples, the RNA samples were extracted and cDNA was synthesized. The MCF-7 cells and SKBR-3 cells were treated with dopamine receptors agonists and antagonists. The MTT test was conducted to identify oxidative and reductive enzymes and to specify appropriate dosage at four concentrations of dopamine and Cabergoline on MCF-7 and SKBR-3 cells. Immunofluorescence staining was done by the use of a mixed dye containing acridine orange and ethidiume bromide on account of differentiating between apoptotic and necrotic cells. Flow cytometry assay was an applied method to differentiate necrotic from apoptotic cells.Results:Sixty seven and thirty three percent of the patients were related to stages II and III, respectively. About sixty three percent of the patients expressed ER, while fifty seven percent expressed PR. Thirty seven percent of the patients were identified as HER-2 positive. All types of D2-receptors were expressed in PBMC of patients with breast cancer and healthy individuals. The expression of the whole dopamine receptor subtypes (DRD2-DRD4) was carried out on MCF-7 cell line. The results of RT-PCR confirmed the expression of DRD2 on SKBR-3 cells, whereas the other types of D2- receptors did not have an expression. The remarkable differences in gene expression rates between patients and healthy individuals were revealed in the result of the Real-time PCR analysis. The over expression in DRD2 and DRD4 genes of PBMCs was observed in the patients with breast cancer at stages II and III. The great amount of apoptosis and necrosis occurred after the treatment of MCF-7 cells by Cabergoline from 25 to 100 µmolL-1 concentrations.Conclusion:This study revealed the features of dopamine receptors associated with apoptosis induction in breast cancer cells. Moreover, the use of D2-agonist based on dopamine receptors expression in various breast tumoral cells could be promising as a new insight of complementary therapy in breast cancer.

scholarly journals Role of Cardiac Biomarkers in the Assessment of Acute Cerebrovascular Accident

2021 ◽  
Vol 12 (01) ◽  
pp. 106-111
Author(s):  
Srinivasan Radhakrishnan ◽  
Swathy Moorthy ◽  
Sudish Gadde ◽  
Krishnaswamy Madhavan
Keyword(s):  
Acute Stroke ◽  
Cerebrovascular Accident ◽  
Troponin I ◽  
Serum Levels ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cardiac Enzymes ◽  
Mean Values ◽  
Creatine Kinase Mb ◽  
The Mean

Abstract Background Stroke (cerebrovascular accident) has for long been a global burden in terms of its morbidity and mortality. Serum levels of cardiac enzymes such as creatine kinase-MB (CK-MB) component, troponin T, and brain natriuretic peptide have been found to be elevated among the patients with stroke and also serve to prognosticate these patients. The serum levels of these enzymes correlate directly to the severity of stroke in these patients. Objective Elevated cardiac enzymes among patients with acute cerebrovascular accidents are not uncommon despite the patients not having any cardiac problems. We aimed to identify the occurrence of elevated cardiac enzymes among patients with acute stroke and their correlation with the severity of stroke. Materials and Methods Our study included 100 patients of acute stroke with no previous history of cardiac ailments. Serum levels of troponin I and CK-MB were analyzed among these patients using enzyme-linked immunosorbent assay method within the first 2 hours of admission. Patients outcome during the hospital stay were analyzed. Stroke severity was assessed using the National Institute of Health Stroke score (NIHS score) and the modified Rankin Score (mRS). The cardiac enzyme levels were correlated with these scores. Results Twenty-eight percent of patients had elevated troponin I, while 72% patients had normal levels with the mean values of 10.36 to 106.54 ng/mL and 0.00 to 0.02 ng/mL, respectively. CK-MB levels were found elevated among 14% patients and normal among 86% patients with mean values of 5.8 to 124.36 and 0.0 to 4.3 ng/mL, respectively. Among the six patients who succumbed to death, three patients had increased troponin I and four had elevated CK-MB. NIHS scores of 21.0357±6.79 and 105.277±5.564 were seen in patients with elevated and normal troponin I, whereas NIHS scores of 20.4285±8.658 and 11.8721±9.273 were seen among patients with increased and normal CK-MB, respectively. The mRS scores were 4.3214±0.367, 2.4305±1.374, 4.2143±1.412, and 2.756±1.749 ng/mL among the patients with elevated and normal troponin I and CK-MB, respectively. Conclusion The mean values of cardiac enzymes troponin I and CK-MB were higher among patients with higher scores of NIHS and mRS. Among them, troponin I was very significant and it may serve as an early biomarker for the severity of stroke and hint on early cardiac evaluation among these patients.

scholarly journals Co60 Vitamin B12 Binding Capacity of Human Leukocytes

Blood ◽  
1962 ◽  
Vol 19 (2) ◽  
pp. 229-235 ◽  
Author(s):  
LEO M. MEYER ◽  
EUGENE P. CRONKITE ◽  
INEZ F. MILLER ◽  
CLAIRE W MULZAC ◽  
IRVING JONES
Keyword(s):  
Vitamin B12 ◽  
Human Serum ◽  
Polycythemia Vera ◽  
Binding Capacity ◽  
Chronic Myelocytic Leukemia ◽  
Nonspecific Binding ◽  
Two Phase ◽  
Intact Cells ◽  
Myelocytic Leukemia ◽  
Neutrophilic Leukocytes

Abstract 1. Mature neutrophilic leukocytes show the highest Co60B12 binding capacity. 2. Less mature granulocytes, "blast" forms and eosinophils have little or no Co60B12 binding capacity. 3. Disintegrated mature leukocytes from chronic myelocytic leukemia and polycythemia vera show higher B12 binding capacity than intact cells. 4. Mature leukocytes from patients with chronic myelocytic leukemia and polycythemia vera show a two-phase B12 curve suggesting specific and nonspecific binding, similar to that observed in human serum. 5. Disintegration products from mature neutrophilic leukocytes probably contribute largely to increased B12 binding capacity of serum in chronic myelocytic leukemia and polycythemia vera.

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