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Biomedical subjects

R W Alexander

Publications and source records attributed to R W Alexander.

At least 271 records · Page 15Linked to original sources
PubMed1969

Diabetes mellitus--current criteria for laboratory diagnosis.

Among the laboratory methods and test procedures used for the diagnosis of diabetes mellitus in clinical practice, the oral glucose tolerance test, if performed properly, remains the cornerstone in the definitive diagnosis of diabetes. The intravenous glucose tolerance or the tolbutamide response tests may be indicated especially when there is a question of proper oral carbohydrate assimilation. The value of the cortisone-glucose tolerance test is still in question. Insulin assays may be helpful in differentiating between the various types of diabetes. Accepted criteria for normal and abnormal values for all tests are tabulated.

Clinical Laboratory Techniques↗
PubMedDate not supplied

Interactions between coronary occlusion and the renin-angiotensin system in the dog.

Studies were carried out in 39 barbiturate-anesthetized dogs to determine whether the renin-angiotensin system is important in control of hemodynamics and coronary flow during myocardial ischemia. Plasma renin activity (PRA) was 2.2 +/- 0.4 ng x ml-l x hr-1 immediately before coronary artery occlusion (CAO) and increased to 3.8 +/- 0.5 (p less than .005) 15 minutes after CAO. In nephrectomized dogs, PRA was 0.76 +/- 0.14 ng x ml-l x hr-1 two hours after nephrectomy and remained unchanged after CAO. In contrast, hemodynamic changes following CAO were similar between nephrectomized and intact dogs: mean arterial pressure fell from 126 +/- 4 pre CAO to 116 +/- 4 mm Hg post CAO (p less than 0.005) in nephrectomized dogs and from 130 +/- 11 to 120 +/- 11 mm Hg (p less than 0.005) in intact dogs. Left atrial pressure rose from 5.4 +/- 0.9 pre CAO to 7.7 +/- 0.9 mm Hg (p less than 0.005) post CAO in nephrectomized from 5.4 +/- 0.9 pre CAO to 7.7 +/- 0.9 mm Hg (p less than 0.005) post CAO in nephrectomized dogs and 6.3 +/- 1.3 to 9.0 +/- 1.8 mm Hg (p less than 0.005) in intact dogs. Heart rate remained unchanged in both groups. In sham-operated dogs without CAO, neither the angiotensin II blocker Saralasin nor the converting enzyme inhibitor Captopril had significant effects on systemic (SVR) and coronary (CVR) vascular resistances. In contrast, in dogs with CAO, these drugs reduced CVR from 1.28 +/- 0.13 mm Hg x ml-1 x min x 100 g heart weight (resistance units = RU) to 0.85 +/- 0.08 RU (p less than 0.05) (Saralasin) 15 minutes after treatment and from 1.17 +/- 0.09 to 0.88 +/- 0.08 RU (p less than 0.025), (Captopril) respectively. However, only Captopril reduced SVR, from 10.7 +/- 1.13 to 8.2 +/- 0.8 RU (p less than 0.025). Both Captopril and Saralasin induced a significant increase in collateral blood flow. Nephrectomy, two hours prior to CAO, significant increase in collateral blood flow. Nephrectomy, two hours prior to CAO, significantly reduced the effect of Captopril on CVR and collateral blood flow while the effect on SVR persisted. Thus the reduction in CVR appears to be an effect of inhibition of the renin-angiotensin system; this system participates in control of CVR during CAO and may limit coronary collateral blood flow.

Animals↗
PubMedDate not supplied

An endothelial cell-smooth muscle cell co-culture model for use in the investigation of flow effects on vascular biology.

Flow and the associated shear stress have been shown to play an active role in the regulation of the structure and function of endothelial cells (EC) in vitro. Although cultured EC subjected to flow exhibit an elongated morphology and a decreased cell growth rate rather like those observed in vivo, there are differences in morphology and growth rate, as well as other characteristics, between in vitro and in vivo EC. This suggests that flow is only one of the many factors affecting EC differentiation in vivo. In this study, a co-culture model system was designed, which includes smooth muscle cells (SMC), a matrix of collagen type I, and a confluent monolayer of EC, and this simplified model of the arterial wall was subjected to a steady, laminar shear stress of 10 and 30 dyn/cm2. Under non-flow conditions, EC exhibited an elongated shape, but with a random orientation. In response to flow, there was an alignment with the direction of flow. This alignment occurred more rapidly at 30 dyn/cm2 than at 10 dyn/cm2. The collagen matrix was found to be primordial in the maintenance of a quiescent endothelium, even in the absence of SMC and flow, suggesting the importance of an organized extracellular matrix (ECM) in the differentiation of cells in vivo.

Animals↗
PubMedDate not supplied

Angiotensin-II-and endothelin-induced protein phosphorylation in cultured vascular smooth muscle cells.

In cultured vascular smooth muscle cells, angiotensin II and endothelin stimulate a variety of intracellular signals, including generation of inositol trisphosphate and diacylglycerol, mobilization of intracellular calcium, and activation of protein kinase C. These latter two events have been shown to mediate the phosphorylation of numerous proteins, but these substrates and the specific pathways mediating their phosphorylation have not been identified in vascular smooth muscle. Angiotensin II (100 nM, 10 min) induced a characteristic pattern of protein phosphorylation, which included the phosphorylation of many proteins, ranging in molecular mass from 20 to 76 kD. Three of these proteins have been identified as vimentin (M(r) 57,000), a specific protein kinase C substrate (M(r) 76,000) and the myosin light chain (M(r) 20,000). The 76-kD protein was one of the most highly phosphorylated proteins after agonist treatment. Endothelin-1 produced an identical pattern of phosphorylation. Five of these substrates were also phosphorylated by phorbol-12-myristate-13-acetate, and 5 were also phosphorylated after treatment with ionomycin. In general, the protein-kinase-C-dependent phosphorylations were sustained, while those mediated by calcium were rapid. Since these experiments were performed in cultured, phenotypically modulated cells stimulated with agents that promote cellular hypertrophy or hyperplasia, this pattern of phosphorylation may be representative of that seen during the growth response.

Angiotensin II↗
PubMedDate not supplied

Characterization of postsynaptic alpha-adrenergic receptors by [3H]-dihydroergocryptine binding in muscular arteries from the rat mesentery.

Alpha-adrenergic receptors are likely to be important determinants of the effects of catecholamines on vascular resistance. To study the alpha-adrenergic receptor in muscular arteries of the type that determine vascular resistance, we characterized and quantitated alpha-adrenergic receptors in a particulate fraction of the highly reactive, richly innervated arteries of the rat mesentery. With the ligand [3H]-dihydroergocryptine ([3H]-DHEC), specific binding (displaceable by 5 microM phentolamine) is saturable. There is a single class of binding sites with a dissociation constant (Kd) of 2.9 nM and a maximal binding capacity of 68 fmoles of [3H]-DHEC per mg of particulate fraction protein. Catecholamines compete for [3H]-DHEC binding stereospecifically and with the alpha-adrenergic potency series of (-)epinephrine greater than (-)norepinephrine greater than (-)isoproterenol. Binding is rapid (t 1/2 less than or equal to 2 mins) and rapidly reversible (t 1/2 less than or equal to 2 mins). Inhibition of [3H]-DHEC binding by the alpha-adrenergic antagonist phentolamine (Kd = 3.0 nM) is much greater than by the beta-adrenergic antagonist propranolol (Kd = 8200 nM). The alpha 1-selective antagonist prazosin (Kd = 63 nM) is 20 times more potent in competing for [3H]-DHEC binding than is the alpha 2-selective antagonist yohimbine (Kd = 1250 nM), thus suggesting that the alpha-adrenergic receptor identified is predominantly of the alpha 1 subtype that is responsible for vascular smooth muscle contraction. This extension of radioligand binding techniques to highly innervated muscular arteries of the type contributing to vascular resistance will allow the study of the role of the vascular alpha-adrenergic receptor in various physiologic states and models of hypertension.

Adrenergic alpha-Antagonists↗
PubMedDate not supplied

Regulation of alpha 1-adrenergic receptor-coupled calcium flux in cultured vascular smooth muscle cells.

To study the role of the alpha-adrenergic receptor (AAR) pathway in the regulation of catecholamine-induced vascular contraction, we developed methods for the direct evaluation of AAR and AAR-coupled calcium efflux in cultured vascular smooth muscle cells derived from the rabbit aorta by enzymatic dissociation. AAR were characterized by the binding of the alpha 1- and alpha 2-selective radioligands [3H]-prazosin and [3H]-yohimbine, respectively. Norepinephrine-stimulated efflux of 45calcium from preloaded cells was measured as an index of AAR-mediated calcium flux. The [3H]-prazosin binding was of high affinity (Kd = 0.15 nM), saturable (Bmax = 75-125 fmol/mg protein), and competed for by agonists and antagonists in the order expected for an alpha 1 receptor. There was no specific binding of [3H]-yohimbine. Norepinephrine-stimulated 45Ca efflux was concentration-dependent (EC50 congruent to 100 nM), and potently blocked by prazosin (IC50 congruent to 0.1 nM), but not yohimbine (IC50 greater than 100 nM), indicating, together with the binding data, that norepinephrine-stimulated 45Ca efflux in this system is alpha 1 mediated. Following treatment of cells with 1-norepinephrine (0.1 nM) for 24 hours, the density of AAR and maximal norepinephrine-stimulated 45Ca efflux were decreased by 72% +/- 14% and 91% +/- 9%, respectively. This cultured cell system offers several advantages for study of the mechanisms of coupling and regulation of alpha-adrenergic receptors in vascular smooth muscle.

Animals↗
PubMedDate not supplied

Angiotensin increases cytosolic free calcium in cultured vascular smooth muscle cells.

We used the calcium-sensitive fluorescent indicator quin 2 to monitor changes in cytosolic free calcium concentration ([Ca2+]i) associated with angiotensin II receptor activation in cultured vascular smooth muscle cells isolated from rat aorta. Resting [Ca2+]i in unstimulated vascular smooth muscle cells was 198 +/- 7 nM. Angiotensin II induced concentration-dependent rapid increases in [Ca2+]i (threshold congruent to 10(-11) M; effective concentration, 50% congruent to 5 X 10(-10) M; maximum congruent to 10(-8) M); the rate of increase in [Ca2+]i also appeared to be concentration dependent. The angiotensin II-induced changes were completely blocked by the angiotensin II receptor antagonist [Sar1, Ile8]-angiotensin II. In the presence of extracellular calcium, 10(-8) M angiotensin II induced an increase in [Ca2+]i that reached peak values of five to six times the resting levels within 15 seconds, followed by a gradual decline to a plateau at two to three times the resting level. When EGTA was added to chelate external calcium, the angiotensin II-induced increases in peak [Ca2+]i were attenuated and the plateau phase was abolished. These data show that (1) quin 2 can be used in cultured vascular smooth muscle cells to study changes in calcium homeostasis induced by angiotensin II, and (2) angiotensin II acts on cultured vascular smooth muscle cells to cause a rapid increase in [Ca2+]i that appears to depend on both the mobilization of intracellular calcium and the influx of extracellular calcium.

Aminoquinolines↗