Estrogen reduces proliferation and agonist-induced calcium increase in coronary artery smooth muscle cells.

Epidemiological evidence and estrogen replacement studies suggest that estrogen has a protective effect on the cardiovascular system against coronary artery disease. Vascular smooth muscle (VSM) cell replication has been shown to play a causative role in the pathogenesis of atherosclerosis. Therefore, in this study, we investigated the effect of chronic treatment of cultured guinea pig coronary artery VSM cells with physiological concentrations of 17beta-estradiol (E2) on thymidine incorporation, cell proliferation, and bradykinin-stimulated cytosolic calcium concentration ([Ca2+]i). Bradykinin at physiological concentrations causes contraction of endothelium-denuded guinea pig coronary artery rings in a concentration-dependent manner. VSM cells were first treated with low doses of E2 (10 pg/ml) for 1-2 days followed by treatment for 4-6 days with 50 pg/ml of E2, a concentration similar to that found in pregnancy. Using these protocols, we consistently observed the presence of E2-receptor mRNA in VSM cells by a ribonuclease protection assay. Fetal calf serum-stimulated [3H]thymidine incorporation was significantly reduced (P < 0.05) in E2-treated cells compared with untreated control cells. Similarly, E2 treatment significantly inhibited fetal calf serum-stimulated VSM cell proliferation compared with untreated control cells (P < 0.05). We also tested the hypothesis that E2 treatment attenuates agonist-stimulated [Ca2+]i in VSM cells because acute E2 treatment has been shown to produce relaxation of precontracted isolated coronary artery preparations. E2 treatment of VSM cells resulted in a significant decrease in bradykinin-stimulated [Ca2+]i compared with untreated cells (P < 0.05). In conclusion, our data demonstrate that estrogen at physiological concentrations directly regulates coronary VSM cell function.

Vascular effects of metformin. Possible mechanisms for its antihypertensive action in the spontaneously hypertensive rat.

Metformin, an antidiabetic agent, potentiates insulin action and reduces insulin resistance. We examined the antihypertensive effects and vascular effects of metformin in spontaneously hypertensive rats (SHR). Wistar-Kyoto normotensive (WKY) and SHR were injected with metformin (100 mg/kg) or saline subcutaneously twice daily for 4 weeks. Blood pressure was recorded by a tail-cuff plethesmographic method. Metformin treatment significantly attenuated (P < .05) the increase in blood pressure in metformin treated SHR versus untreated control SHR. At the end of the experimental period of 4 weeks, metformin-treated SHR had a mean blood pressure that was 34 mm lower than that of untreated SHR. Metformin treatment had no significant effect on blood pressure in WKY rats. Treatment of SHR aortic smooth muscle (SM) cells with metformin (2 micrograms/mL) for 24 h significantly decreased (P < .05) arginine vasopressin- and thrombin- stimulated increase in [Ca2+]i. However, metformin treatment did not have a significant effect on the basal [Ca+]i. Incubation of SHR aortic SM cells with OH-L-arginine (25 to 100 mumol/L) for 24 h increased nitrite production in a dose dependent manner. Metformin (5 micrograms/mL) treatment of SM cells increased nitrite production at all concentrations of OH-L-arginine; however, differences were significant (P < .05) only at 25 and 50 mumol/L OH-L-arginine. These results suggest that metformin may be decreasing arterial pressure in the SHR, at least in part, by attenuating the agonist-stimulated [Ca2+]i response in SHR vascular smooth muscle cells.

Phylogenetic relationships among members of the class Mollicutes deduced from rps3 gene sequences.

A gene for a ribosomal protein, rps3, was amplified by PCR and sequenced from representatives of the class Mollicutes. Alignments of the deduced amino acid sequences allowed the construction of a phylogeny that is consistent with the phylogenetic trees created from 5S and 16S rRNA comparisons, including the position of the former Acholeplasma florum on the Mycoplasma branch, rather than with the classical Acholeplasmataceae. Additional confirmation of the phylogeny comes from the deduction that the UGA triplet encodes tryptophan in the rps3 gene from Mesoplasma florum, as it does in the mycoplasmas and spiroplasmas. The sequence data from Acholeplasma axanthum 743 and Acholeplasma sp. strain J233 allow refinements to the phylogenetic tree within the Acholeplasmataceae, providing evidence that the sterol requirement of Anaeroplasma abactoclasticum (order Anaeroplasmatales) is a derived trait. It was also evident that the nonhelical plant-pathogenic members of the class Mollicutes, referred to as mycoplasma-like organisms or phytoplasmas, are more closely related to the true acholeplasmas (Acholeplasma laidlawii and strain J233) than to other members of the Mollicutes.