Substrate specificity of CTP:phosphocholine cytidylyltransferase.

The specificity of CTP:phosphocholine cytidylyltransferase from rat liver for phosphorylated bases has been investigated. The apparent Km for phosphocholine was 0.17 mM. As the number of methyl substituents on the phospho-base decreased, the apparent Km increased: 4.0 mM for phosphodimethylethanolamine, 6.9 for phosphomonomethylethanolamine and 68.4 for phosphoethanolamine. The Vmax for the reaction was similar for phosphocholine (12.6 mumol/min per mg protein), phosphomonomethylethanolamine (13.5 mumol/min per mg protein) and phosphoethanolamine (9.2 mumol/min per mg protein). When phosphodimethylethanolamine was the substrate, the Vmax was 3-fold higher (40.3 mumol/min per mg protein). Phosphoethanolamine, phosphomonomethylethanolamine and phosphodimethylethanolamine were competitive inhibitors of the cytidylyltransferase when phosphocholine was used as substrate with Ki values of 18.5 mM, 9.3 mM and 1.5 mM, respectively. The results show that the cytidylyltransferase is highly specific for phosphocholine.     

Mechanism of hyperoxia-induced pulmonary vascular paralysis: effect of antioxidant pretreatment

We designed experiments using isolated rabbit lungs to determine the effect of hyperoxia on the pulmonary vasoconstriction caused by the infusion of the lipid peroxide tert-butyl hydroperoxide (t-bu-OOH), which produces vasoconstriction by stimulating the pulmonary synthesis of thromboxane. Exposure to 48-60 h of 100% O2 at 1 ATA markedly reduced the increase in pulmonary artery pressure caused by t-bu-OOH infusion. We also investigated whether the mechanism for the attenuated vasoconstriction was due to altered production of arachidonate mediators or oxidant-induced damage to the contractile mechanism. In addition to infusing t-bu-OOH, which selectively stimulates thromboxane production, we also infused Intralipid, an esterified fatty acid emulsion that stimulates production of both thromboxane and prostacyclin. These experiments were done to study the effect of hyperoxia on prostacyclin synthesis. To determine if antioxidant therapy would prevent the changes in mediator production and vascular reactivity caused by hyperoxia, we pretreated animals with the antioxidants butylated hydroxyanisole (BHA) or vitamin E. The lack of vascular reactivity to t-bu-OOH was not due to a decrease in thromboxane synthesis or an increase in prostacyclin synthesis. Hyperoxia did not affect thromboxane synthesis during basal conditions or after stimulation of synthesis by t-bu-OOH. 100% O2 also did not effect the basal synthesis of prostacyclin by the lung. Hyperoxia did, however, markedly reduce prostacyclin synthesis when it was stimulated by Intralipid infusion. Antioxidant pretreatment did not reverse the inhibition of prostacyclin synthesis but did prevent the loss of vascular reactivity caused by hyperoxia. Thus hyperoxia causes vascular paralysis through oxidant-induced injury to the pulmonary vasculature.     

Mechanisms by which endothelin 1 induces pulmonary vasoconstriction in the rabbit.

To investigate the mechanisms by which endothelin 1 (ET-1) causes pulmonary vasoconstriction, we studied the effect of synthetic ET-1 on pulmonary vascular tone in the buffer-perfused isolated rabbit lung. In nanomolar concentrations (1.2-8 nM), ET-1 causes a dose-dependent increase in pulmonary arterial pressure that persists for greater than or equal to 1 h (increase in pressure 19 +/- 2 mmHg with ET-1 vs. 2 +/- 1 with vehicle, P less than 0.0001). Reduction of calcium availability with verapamil, cadmium, or a calcium-free buffer significantly blunts the increase in pressure caused by ET-1. Pretreatment with a calcium-free buffer plus the chelator ethylene glycol-bis(beta-aminoethyl ether)-N,N,N', N'-tetraacetic acid (EGTA) completely eliminates the vasoconstriction. Three different inhibitors of protein kinase C, phloretin, staurosporine, and dihydrosphingosine, significantly diminish the response to ET-1. Indomethacin and a thromboxane synthase inhibitor partially decrease the response to the highest concentration of ET-1. Isoproterenol and dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP) are significantly more effective in preventing the vasoconstriction caused by ET-1 than are nitroprusside and guanosine 5'-cyclic monophosphate (cGMP) analogues. ET-1 in doses of 1.2-8 nM is a potent pulmonary vasoconstrictor in the isolated rabbit lung. ET-1 appears to cause pulmonary vasoconstriction by increasing calcium entry and by activating protein kinase C. Vasodilators that increase cAMP are substantially more effective in preventing the increase in pressure than are drugs that increase cGMP.     

Myelin basic protein purified on an ion-exchange continuous polymer bed in the presence of ethylene glycol and salt possesses activity againstp-nitrophenyl acetate

In this paper we describe a fast and mild method based on the use of a unique cation exchanger and buffers containing ethylene glycol and salt for the purification of the myelin basic protein (MBP; MW 18.5 kDa). MBP thus purified hydrolyses catalytically p-nitrophenyl acetate. This esterase activity facilitates not only the purification of MBP but also indicates that probably it is in its native state, i.e. there is a good chance that the purified molecules are structurally and chemically identical. This is a prerequisite to obtain crystals appropriate for x-ray diffraction and other studies.Abbreviations used MBP myelin basic protein - MW molecular weight - kDa kilo Dalton - octyl-POE n-octylpolydisperse oligooxyethylene - CHAPS 3-3-cholamidopropyl dimethylammonio-1-propane-sulfonate - CTAB cetyltrimethylammonium bromide - SDS sodium dodecyl sulfate - SDS-PAGE polyacrylamide gel electrophoresis in the presence of SDS - G 3707 heptaoxyethylene lauryl ether - TWEEN-20 polyoxyethylenesorbitan-monolaurat - EDTA ethylenediaminetetraacetic acid - HEPES N-(2-hydroxyethyl)-piperazine-N-(2-ethanesulfonic acid)     

Salinity induced changes in the reproductive physiology of wheat plants

The effect of salinity on reproductive physiology of wheat wasinvestigated. One set of wheat plants was subjected to increasingsalt levels up to a certain concentration, whereas another setwas given the same concentration in a single application. Theformer was called "gradual" and latter "shock" treatment. Theireffects on pollen viability, germination and activity of starchsynthetase were studied. Gradual treatment seemed to reducethe toxic effects of salts on the viability of pollen grainsand their germination. Seeds obtained from the two sets weregerminated in the same salinities in which their plants hadbeen growing, and the results were compared with those of seedsobtained from control plants growing in a non-saline medium.The seeds of plants from the gradual treatment were better suitedfor germination on a saline medium than those from the shocktreatment or the control group. Salt treatment also increasedthe activity of starch synthetase at the midmilky stage in developinggrains. This phenomenon was considered essential for synthesisof starch in a saline environment. The increase in Na⁺ and Cl^– and decrease in K⁺ contentsof wheat grains in both treatments suggest that the effect ofsalinity on the physiological phenomenon studied is due to changesin the ionic content of the plants. ¹ In partial fulfilment of a Ph.D. degree from the Universityof Karachi, Pakistan. ² Professor of Botany, Director of Research Projects, Head,Plant Physiology Section, University of Karachi, Pakistan. (Received July 11, 1977; )     

The fauna of benthic sediments from the organically enriched Oslofjord,Norway

The macrobenthic fauna of the organically enriched Oslofjord was sampled in Sept.–Nov. 1977. Seventy-six stations were sampled using a stratified random plan. A total of 146 species were identified, dominated by polychaetes, molluscs and echinoderms. The faunal data were analysed by a variety of methods including diversity indices, log-normal distribution of individuals among species, factor analysis and numerical classification. The primary aim was the detection of pollution-induced disturbance and to delimit the extent of pollution on the benthic fauna of Oslofjord. All of the methods used showed a gradient from heavy pollution at the innermost part of the fjord where few species occurred, together with high dominance and low diversity, to normal unpolluted conditions at the Drøbak sill with high species diversity. From the classification analysis a combination of both site and species data revealed a trend in species groupings which were broadly similar to data from other Scandinavian fjords and from Scotland. Hydrographical and biological data taken at the turn of the century and in the 1960's compared with the present data show declining conditions in Oslofjord, due primarily to organic enrichment combined with a naturally poor water exchange which leads to stagnation of the water mass.     

Coassemble dopamine and GHK tripeptide into fluorescent nanoparticles for pH sensing

Fluorescent nanostructures have been widely applied to biomedical researches and clinical diagnosis such as biolabeling/imaging/sensing and have even acted as therapy reagents. Peptide‐based fluorescent nanostructures attract recent interest from biomedical researchers. Inspired by the natural existence of GHK‐Cu complex with a growth factor‐like effect in human blood, here we have developed a novel approach for designing nanosensors through the co‐assembling of two kinds of biomolecules. By making best use of both π‐π stacking between carbon rings and the easy‐oxidation property of an important transmitter molecule, dopamine (DA), we successfully built up a supersensitive and robust fluorescent pH nanosensor by co‐assembling oxidized DA (DA_ox) with a tripeptide GHK. The GHK‐DA_ox nanostructures have a quantum yield of 20.82%, which might be the brightest one among all the current co‐assembling structures merely through unmodified biomolecules. We envision this approach could open a new avenue for not only hybrid nanostructure construction, but also may inspire the bioengineering of in vivo luminescent probes.