Beat U. Raess

The Teaching of Complementary and Alternative Medicine in U.S. Medical Schools: A Survey of Course Directors.

Purpose The number of U.S. medical schools offering courses in complementary and alternative medicine (CAM) has risen sharply in recent years. This study gauged the current state of CAM instruction by gathering details about the specific topics being taught and the objectives behind the instruction. Method Data were collected from questionnaires mailed to 123 CAM course directors at 74 U.S. medical schools. Results Questionnaires were returned by 73 course directors at 53 schools. The topics most often being taught were acupuncture (76.7%), herbs and botanicals (69.9%), meditation and relaxation (65.8%), spirituality/faith/prayer (64.4%), chiropractic (60.3%), homeopathy (57.5%), and nutrition and diets (50.7%). The amounts of instructional time devoted to individual CAM topics varied widely, but most received about two contact hours. The “typical” CAM course was sponsored by a clinical department as an elective, was most likely to be taught in the first or fourth year of medical school, and had fewer than 20 contact hours of instruction. Most of the courses (78.1%) were taught by individuals identified as being CAM practitioners or prescribes of CAM therapies. Few of the courses (17.8%) emphasized a scientific approach to the evaluation of CAM effectiveness. Conclusion A wide variety of topics are being taught in U.S. medical schools under the umbrella of CAM. For the most part, the instruction appears to be founded on the assumption that unconventional therapies are effective, but little scientific evidence is offered. This approach is questionable, especially since mainstream medicine owes much of its success to a foundation of established scientific principles.

Verapamil, diltiazem and nifedipine interactions with calmodulin stimulated (Ca2++Mg2+)-ATPase

The functional interactions of the three prototype Ca2+ antagonists, verapamil, diltiazem and nifedipine, were examined in relation to the calmodulin regulated plasma membrane Ca2+ pump ATPase. For this we used low ionic strength derived, calmodulin depleted, human red cell ghost membranes. Exogenously added calmodulin activated basal (Ca2+ + Mg2+)-ATPase in a concentration-dependent manner. Half-maximal activation by 6 nM calmodulin was antagonized by 10(-3) M verapamil and 10(-3) M diltiazem 25.1 and 12.1% respectively. The inhibition appeared to be specific for calmodulin activation since basal activity was not affected by these agents. Nifedipine had no effects on basal or calmodulin stimulated (Ca2+ + Mg2+)-ATPase activity. Unlike dihydropyridine modulation of verapamil and diltiazem binding at high affinity channel sites, nifedipine in this system did not alter the inhibitory responses of verapamil and diltiazem. The calmodulin directed antagonism of the two drugs was shown to be strictly additive over a full range of calmodulin concentrations and appeared to change predominantly the Vmax and, to a lesser degree, the affinity of calmodulin for the (Ca2+ + Mg2+)-ATPase. It is concluded that this model system provides evidence for additional functional discrepancies among the various classes of Ca2+ antagonists.

Calmodulin-stimulated plasma membrane (Ca2+ + Mg2+)-ATPase: inhibition by calcium channel entry blockers.

Calcium channel entry blockers representing different structural classes were studied for their effects on human erythrocyte basal and calmodulin-stimulated (Ca2+ + Mg2+)-ATPase. Effects on the activity of (Mg2+)-ATPase and (Na+ + K+)-ATPase were also assessed. Of the four Ca2+ entry blockers tested, only verapamil and diltiazem specifically inhibited the calmodulin-stimulated (Ca2+ + Mg2+)-ATPase activity, the basal enzyme activity being unaltered by these drugs. Other membrane-associated ATPases were not affected. Calmodulin concentration effect curves showed the inhibition by verapamil (10(-3) M) and diltiazem (10(-3) M) to be non-competitive. This concentration inhibited the calmodulin-dependent increment (5.1 nM calmodulin) of the ATPase activity by 35 and 36% respectively. Similarly, both drugs inhibited the Ca2+-activation process of calmodulin-stimulated activity in a non-competitive manner, decreasing Vmax by 23 and 17% respectively. Basal (Ca2+ + Mg2+)-ATPase activity was not affected by verapamil or diltiazem at any calcium concentration. In contrast, cinnarizine non-specifically inhibited all four membrane ATPases including calmodulin-stimulated (Ca2+ + Mg2+)-ATPase activity at concentrations above 3 X 10(-6) M. Nifedipine was without effect on any of the four membrane ATPases. From this we conclude that certain calcium channel entry blockers can inhibit calmodulin-regulated plasma membrane Ca2+-pump ATPase. Therefore, this identifies an additional functional low affinity receptor in the plasma membrane for some of the calcium channel entry blockers.

Effects of repeated doses of azapirones on rat brain 5-HT1A receptors and plasma corticosterone levels

Chronic buspirone or ipsapirone (3 mg/kg, twice daily) administration to rats for 10 days decreased the sensitivity of inhibition of single-unit activity of serotonergic dorsal raphe neurons to a challenge by each drug. The ED50 for buspirone was increased from 0.1 mg/kg to 1.8 mg/kg, and the ED50 for ipsapirone was increased from 0.7 mg/kg to 1.2 mg/kg. The binding properties (Kd and Bmax) of [3H]8-OH-DPAT to membranes of cerebral cortex and hippocampus were unaffected by chronic administration of either buspirone or ipsapirone. Chronic buspirone or ipsapirone administration increased the tolerance of the hypothalamic-pituitary-adrenal axis (HPAA) following a challenge by each drug. The ED50 for elevation of plasma corticosterone levels was increased from 4.0 mg/kg to 7.6 mg/kg for buspirone and 6.2 mg/kg to 8.0 mg/kg for ipsapirone. Chronic buspirone administration decreased the basal activity of the HPAA by 63%. Chronic buspirone administration did not alter the plasma corticosterone response of the HPAA to a 1-min episode of rotational stress. (Mg2+)-ATPase, (Na+ + K+)-ATPase, (Ca2+ + Mg2+)-ATPase and calmodulin-stimulated (Ca2+ + Mg2+)-ATPase activities of erythrocyte plasma membrane were unaffected by either chronic or acute buspirone treatment, or by the addition of the drug to the in vitro assay systems.

Inhibition of calmodulin-stimulated (Ca2+ + Mg2+)-ATPase activity by dimethyl sulfoxide

Membrane-bound (Ca2+ + Mg2+)-ATPase activity from human erythrocyte white ghosts in the calmodulin-activated state was inhibited by DMSO in concentrations of 3% (v/v) and above. At 10%, DMSO inhibited calmodulin activation by 47.7%, while basal, calmodulin-independent (Ca2+ + Mg2+)-ATPase and (Mg2+)-ATPase activity remained unaffected. (Na+ + K+)-ATPase activity was also reduced but exhibited a greater IC50. Concentration-effect analyses showed the inhibition by 10% DMSO to be a reversible, non-competitive effect with regard to calmodulin, Ca2+, and substrate activation. Calmodulin-stimulated processes may be more susceptible to inhibition by DMSO than related enzymatic catalysis, and thus may help explain the multitude of reported cellular events caused by the solvent. Furthermore, DMSO affected membrane-associated enzymatic mechanisms opposite to those reported for purified enzyme outside its native membrane environment.

Intracellular Ca 2+ Homeostatic Regulation and 4-Hydroxynonenal-Induced Aortic Endothelial Dysfunction

The aldehydic lipid peroxidation product 4-hydroxynonenal (HNE) is known to compromise erythrocyte passive Ca2+ permeability and to irreversibly inhibit the plasma membrane (Ca2+ + Mg2+)-ATPase and Ca2+-transport. To measure the effects of HNE on passive and active Ca2+ transport in endothelial cells, we first characterized 45Ca2+ uptake and efflux in cultured porcine aortic endothelial cells (PAEC). PAEC exchanged 45Ca2+ to a cumulative near-isotopic equilibrium of about 4.5 pmole 45Ca2+/10(6) cells in 120 min at 37 degrees C. This Ca2+ pool was diminished by thapsigargin, cyclopiazonic acid, oligomycin B, and sodium azide. In contrast, ouabain enhanced Ca2+ uptake capacity from 5.17 to 5.77 pmole/10(6) cells. Accumulated 45Ca2+ was extruded at rate of 8.7 fmole 45Ca2+/10(6) cells/min or shunted rapidly by the ionophore A23187. HNE increased total 45Ca2+ accumulation in a time- and concentration-dependent manner by as much as 562% with an EC50 of 64.0 wM. Concomitant morphological analysis of PAEC revealed vacuolization, nuclear swelling, cell shrinking, and cell detachment. Initial structural changes, such as vacuolization, began well before any changes in Ca2+ accumulation were observed. These functional and morphological changes indicate that HNE significantly increases intracellular Ca2+ accumulation in vascular endothelium, which may explain the cytotoxic effects associated with HNE exposure and provide further evidence that atherogenic effects of HNE may, in part, be caused by disturbances in Ca2+ homeostasis.

Calcium-stressed erythrocyte membrane structure and function for assessing glipizide effects on transglutaminase activation.

Abstract A proposed mechanism of action of hypoglycemic sulfonylureas is the prevention of transglutaminase-mediated endocytosis of insulin receptors. When activated by high levels of intracellular calcium, transglutaminase (TG) catalyzes the cross-linking of intracellular proteins to membrane proteins and modifies membrane structure and function. This study examined the effects of the sulfonylurea glipizide on TG activity in an erythrocyte model by assessing various membrane ATPase activities and high molecular weight protein polymer formation using sodium dodecyl sulfate-polyacryl-amide gel electrophoresis. To activate TG, red blood cells were exposed to 1 mM intracellular Ca2+ using 10-5 M Ca2+-ionophore A23187. In Ca2+-stressed cells, calmodulin stimulation (0.1 μg/ml) of (Ca2+ + Mg2+)-ATPase was decreased to 21.2% of control activity. Increasing concentrations of calmodulin (0.1–3.0 μg/ml) could not overcome the inhibitory effects of TG on the (Ca2+ + Mg2+)-ATPase in Ca2+-stressed cells with or without glipizide. An increased Ca2+ sensitivity of calmodulin-independent (Ca2+ + Mg2+)-ATPase due to Ca2+ stress was seen in all Ca2+-stressed cells even in the presence of 1 mM glipizide. Structural changes were observed in the form of high molecular weight polymer formation. Cells exposed to high Ca2+ and glipizide (3 × 10-5–10-3 M) showed no improvement in ATPase activity or protection from protein cross-linking compared with cells without the drug. We conclude that in this model glipizide fails to inhibit TG induced protein cross-linking and does not prevent the decrease in (Ca2+ + Mg2+)-ATPase activation in Ca2+-stressed red blood cells. This finding considerably weakens the proposal that sulfonylureas act by inhibiting TG activity.

Regulation of Rabbit Erythrocyte Ca2+-Pump Sensitivity to Calmodulin in Experimental Hyperlipidemia

Abstract Intracellular free calcium activity is in part determined by a calmodulin-regulated plasma membrane Ca2+-pump. Since changes in Ca2+ permeability have been implicated in atherosclerotic plaque formation, we initiated a lipid hyperalimentation protocol during which we measured various erythrocyte calcium flux parameters and early atheroma development. Adolescent New Zealand White rabbits were fed a diet with 0.5% cholesterol and 2.5% lard over a 3-month period. Plasma cholesterol and triacylglycerols increased on average 18.7- and 13.9-fold respectively, while erythrocyte membrane cholesterol content decreased 18% and total phospholipids by 54%. After 3 months of lipid hyperalimentation, 22% of the aortic arch was covered with large, early-stage, raised atheroma. Basal and calmodulin-activated (Ca2+ + Mg2+)-ATPase activities in erythrocyte membranes increased by 31% and 123%, respectively at 2 months, with a concomitant increase in calmodulin affinity (Km ) from 15.6 to 4.2 nM. These differences were transient on account of changes in the control animals which exhibited a slowly developing sensitivity to calmodulin during maturation. Basal Ca2+ transport and passive Ca2+ permeability increased about 7-fold during the hyperlipidemic phase. This suggests that overt hyperlipidemia, leading to atherosclerotic plaque development, alters plasma membrane Ca2+ regulatory mechanisms including passive Ca2+ permeability. The changes in enzymatic function, membrane composition, and Ca2+ permeability seen in this red cell model system may be a reflection of early changes in cells that are directly involved in the development of atherosclerotic plaques.