Reduction in number of sarcolemmal KATP channels slows cardiac action potential duration shortening under hypoxia

The cardiovascular system operates under demands ranging from conditions of rest to extreme stress. One mechanism of cardiac stress tolerance is action potential duration shortening driven by ATP-sensitive potassium (K_ATP) channels. K_ATP channel expression has a significant physiologic impact on action potential duration shortening and myocardial energy consumption in response to physiologic heart rate acceleration. However, the effect of reduced channel expression on action potential duration shortening in response to severe metabolic stress is yet to be established. Here, transgenic mice with myocardium-specific expression of a dominant negative K_ATP channel subunit were compared with littermate controls. Evaluation of K_ATP channel whole cell current and channel number/patch was assessed by patch clamp in isolated ventricular cardiomyocytes. Monophasic action potentials were monitored in retrogradely perfused, isolated hearts during the transition to hypoxic perfusate. An 80–85% reduction in cardiac K_ATP channel current density results in a similar magnitude, but significantly slower rate, of shortening of the ventricular action potential duration in response to severe hypoxia, despite no significant difference in coronary flow. Therefore, the number of functional cardiac sarcolemmal K_ATP channels is a critical determinant of the rate of adaptation of myocardial membrane excitability, with implications for optimization of cardiac energy consumption and consequent cardioprotection under conditions of severe metabolic stress.     

Direct measurement of spatial autocorrelation at the community level in four plant communities

Abstract. Four sites were sampled to determine spatial autocorrelation in vegetation at the community level. All were in western New Zealand, but on different substrates and of different physiognomy: a terrace forest, a floodplain forest, a mire and the middle of a logging road. In ‘dissimograms’the four communities all showed steady increases in dissimilarity with distance, but with shoulders in the curve for some sites, which could be related to plant morphology. Most of the increase in dissimilarity occurred over very short distances: less than 0.5 m in the forests, less than 1 m in the mire and less than 2 m in the road. Separate analyses of the woody and herbaceous guilds in the floodplain forest showed that herbaceous dissimilarities remained low at distances up to 20 m, probably because of clonal structure in some species. The mire showed low overall dissimilarity, which is attributed to the uniform substrate and the small species pool. Simulations showed that the approach is capable of indicating structure when it is present. Although the dissimogram was clearest when analysing a simulated grid of patches, other types of simulated patchiness showed dissimograms that were clearly distinguishable from those obtained from the vegetation studied. The almost continuous rise in dissimilarity with distance found in the four sites offers no support to the Hierarchy theory, fitting much more closely the alternative Continuum theory.     

Production of Highly Labeled Adenoviruses

A method is described for increasing the incorporation of radioactive thymidine into adenovirus deoxyribonucleic acid by the use of amethopterin. In addition, a modified procedure is presented for the preparation of highly purified adenoviruses. This procedure, which employs enzymatic digestion of cellular debris, obviates the necessity for fluorocarbon treatment of crude virus suspensions, and routinely provides excellent recovery of virus.     

Radium in the Suwannee River and estuary

A two-year study of radium in the Suwannee River has shown that groundwater discharge, via springs, is a very important source of radium both to the river and to offshore Gulf of Mexico waters. Dissolved radium is maintained within relatively narrow limits in the river by uptake into suspended particles. In the estuary, dissolved radium versus salinity profiles show distinctive nonconservative behavior with radium in significant excess of its linear mixing value at mid-salinities. Unlike the situation in many other estuaries, however, desorption of radium from particles cannot account for most of the observed excess. Thus, the anomalously high radium characteristic of much of the west Florida shelf apparently does not have a riverine source. Direct effusion of high-radium groundwater into these coastal waters is thought to be the major supplier of radium, and perhaps other elements as well.     

Phorbol myristate acetate inhibits growth in S49 cells: isolation of resistant variants

We have used S49 mouse lymphoma cells to study phorbol ester effects on growth. Treatment of wild-type (wt) cells with phorbol 12-myristate 13-acetate (PMA) results in growth arrest within 72 hr. We have selected variants that are resistant to PMA-induced growth arrest, based on a selection in the presence of 10 nM PMA. We have characterized one of these variants, termed 21.1, in detail. The 21.1 and wt cells contain similar levels of protein kinase C (PKC) as determined by [3H]phorbol 12,13-dibutyrate ([3H]PDBu) binding. Treatment of both wt and 21.1 cells with PMA results in translocation of PKC to the membrane, suggesting that the coupling between PKC and an immediate biological response is intact. PMA treatment leads to the phosphorylation of many similar proteins in wild-type and 21.1 cells. However, in the 21.1 cells there is a prominent substrate of approximately 70 kilodaltons (kD) which is no longer phosphorylated after PMA treatment. In wild-type cells ornithine decarboxylase (ODC) activity and mRNA levels are decreased within 1 hr of PMA treatment. Likewise, ODC levels are decreased in the 21.1 cells after exposure to PMA even though PMA only slightly modulates the growth of these cells. The 21.1 cells represent a unique line with a dominant phenotype in which ODC expression is uncoupled from the growth state of the cell. These cells may represent a good model system in which to examine the steps involved in phorbol ester growth regulation in S49 cells.     

An endothelial growth-stimulating factor from salivary glands

Bovine parotid gland and mouse submaxillary gland yield substances of apparent molecular weight 86000 and 80000, respectively which, when injected into either newborn or adult mice induce a hypertrophic growth response from endothelium in numerous organs. The hypertrophic endothelium thus induced closely resembles the endothelium formed in the blood islands of early embryos.     

Rabbit and human liver contain a novel pentacyclic triterpene ester with acyl-CoA: cholesterol acyltransferase inhibitory activity

Acyl-coenzyme A (CoA):cholesterol acyltransferase (ACAT) catalyzes the intracellular fatty acid esterification of cholesterol and is thought to play a key role in lipoprotein metabolism and atherogenesis. Herein we describe the purification and characterization of a novel pentacyclic triterpene ester from rabbit liver that has ACAT inhibitory activity. The inhibitor was purified by a combination of silicic acid chromatography and preparative thin layer chromatography. The compound inhibited both rabbit and rat liver microsomal ACAT activity with an IC50 = 20 microM. The lipid did not inhibit fatty acid incorporation into triglycerides, diglycerides, monoglycerides, or phospholipids nor did it inhibit plasma lecithin:cholesterol acyltransferase activity. However, rat liver microsomal acyl-CoA:retinol acyltransferase activity was inhibited by the terpene ester. Kinetic data are consistent with a mechanism in which ACAT is inhibited by the compound in an irreversible manner. The subcellular fractionation pattern of both ACAT activity and the ACAT inhibitor were similar in rabbit liver (both were approximately equally distributed in membranes that pelleted at 10,000 X g and 100,000 X g). A lipid with similar properties to the rabbit liver inhibitor was found in many other rabbit tissues, including adrenal and spleen, as well as in human liver. Rat liver did not contain this lipid. Structural analysis by NMR, mass spectrometry, and x-ray crystallography indicated that the rabbit liver inhibitor was a fatty acid ester (mostly stearate) of a pentacyclic triterpene acid. The carbon skeleton of the triterpene moiety is a new member of the olean-12-ene triterpene family. Both the negatively charged carboxylic acid group of the triterpene moiety and the esterified fatty acid group were necessary for the ACAT-inhibitory activity of the triterpene ester. Lastly, we present preliminary data which, together with the structural homology of the rabbit triterpene with known plant compounds, suggest the hypothesis that the triterpene moiety of the rabbit ACAT inhibitor arises from dietary absorption of a plant triterpene.