Inductive electron-withdrawal from ammonium ion headgroups of cationic lipids and the influence on DNA transfection

We have prepared a panel of lipidic ammonium tetrafluoroborate salts that contain trifluoromethyl, trichloromethyl, and methyl groups attached to the headgroup. 19F-NMR analyses of the cationic lipid panel revealed that the differences in electron-withdrawal from the ammonium ion headgroup accounted for differences in ion-pairing. Exchange of the tetrafluoroborate counterion by complexation to DNA-phosphate of a reporter gene enabled us to probe the influence of inductive electron-withdrawal in cationic lipid-mediated DNA transfection. We tested the lipid panel for transfection activity in two cell lines. The results indicate that the inductive effects of electron-withdrawing functionality diminish transfection activity in modest (2-4-fold) increments. The present study suggests that the mechanism whereby poly(alcohol)- or poly(ether)-substituted headgroups improve DNA transfection is not based on electronic activation of the ammonium ion.     

Prenylation of olivetolate by a hemp transferase yields cannabigerolic acid, the precursor of tetrahydrocannabinol

A new enzyme, geranylpyrophosphate:olivetolate geranyltransferase (GOT), the first enzyme in the biosynthesis of cannabinoids could be detected in extracts of young leaves of Cannabis sativa. The enzyme accepts geranylpyrophosphate (GPP) and to a lesser degree also nerylpyrophosphate (NPP) as a cosubstrate. It is, however, specific for olivetolic acid; its decarboxylation product olivetol is inactive as a prenyl acceptor.     

Serum factors releasing serotonin from normal platelets: relation to the manifestations of systemic lupus erythematosus

In 13 patients with systemic lupus erythematosus, globulin fractions of sera from serial bleedings were assayed for the induction of serotonin release from normal platelets. Releasing activity appeared at some time in the sera of seven patients, five of whom had episodes of thrombocytopenia. Conversely, only one patient manifested thrombocytopenia without the occurrence of serotonin releasing activity at any time. In three patients with episodes of thrombocytopenia, increases in serotonin releasing activity temporally coincided with drops in platelet count. These data show that levels of circulating platelet serotonin releasing factor(s) vary in the course of systemic lupus erythematosus and these variations may be inversely related to the platelet count in particular patients.     

Regulation of acetylcholine synthesis: does cytoplasmic acetylcholine control high affinity choline uptake?

When brain synaptosomes are obtained from animals that have been injected intravenously with [2H4]choline 1 minute before being killed, their high affinity [3H] choline uptake is correlated inversely with their acetylcholine content and directly with the rate at which they synthesize [2H4]acetylcholine. The control of such choline uptake by the cytoplasmic acetylcholine concentration is proposed as a mechanism regulating acetylcholine synthesis in cholinergic nerve terminals.     

Analysis of a large pedigree with elliptocytosis, multiple lipomatosis, and biological false-positive serological test for syphilis

Elliptocytosis, multiple lipomatosis, and biological false-postive serological test for syphilis (BFPSTS) were found in a single individual. One hundred eighty relatives were tested for the three diseases: 74 were typed for seven blood group antigens, and 58 were typed for four electrophoretic enzyme markers. Likelihood analysis of the pedigree data confirmed independent dominant inheritance for elliptocytosis and lipomatosis. BFPSTS appears dominant, but the analysis was inconclusive. No linkages were found between any disease gene and any marker gene. Two female pedigree members with BFPSTS developed systemic lupus erythematosus, a finding in agreement with the previously described association. The analysis did not lead to any conclusions about the causal relationship between the two traits.     

Exercise tolerance at 4 and 6 ATA

Seven normal male subjects performed 5-min bicycle exercise ranging from 50-100% maximum oxygen uptake at 4 ATA and three were also studied at 6 ATA. At all pressures, the subjects breathed 0.2 ATA O2 plus nitrogen. All subjects were able to perform maximum work at all pressures. No pressure-dependent variations in heart rate, O2 uptake, or CO2 output were noted. At both 4 and 6 ATA, ventilation was decreased at exercise levels greater than 80% maximum O2 uptake. The magnitude of the decrease was not great, however, and signified only minor CO2 retention. In some instances exercise ventilation closely approached the 15-S maximum breathing capacity and these subjects noted severe dyspnea, possibly due to dynamic compression of large airways. In three subjects, respiratory frequency was measured as well as minute ventilation; this relationship did not change with depth. Subjects performing heavy exercise at 6 ATA noted disturbances of consciousness, presumably due to N2 narcosis.     

Sampling and quantifying invertebrates from drinking water distribution mains

Water utilities in the Netherlands aim at controlling the multiplication of (micro-) organisms by distributing biologically stable water through biologically stable materials. Disinfectant residuals are absent or very low. To be able to assess invertebrate abundance, methods for sampling and quantifying these animals from distribution mains were optimised and evaluated. The presented method for collecting invertebrates consists of unidirectionally flushing a mains section with a flow rate of 1 ms(-1) and filtering the flushed water in two separate flows with 500 microm and 100 microm mesh plankton gauze filters. Removal efficiency from mains was evaluated in nine experiments by collecting the invertebrates removed from the mains section by intensive cleaning immediately subsequent to sampling. Of 12 taxa distinguished, all except case-building Chironomidae larvae (2%) and Oligochaeta (30%) were removed well (51-75%). Retention of invertebrates in 100 microm filters was evaluated by filtering 39 filtrates using 30 microm filters. Except for flexible and small invertebrates such as Turbellaria (13%), Nematoda (11%) and Copepoda larvae (24%), most taxa were well retained in the 100 microm filters (53-100%). During sample processing, the method for taking sub-samples with a 10 ml pipette from the suspension of samples with high sediment concentrations was found to perform well in 75% of the samples. During a 2-year national survey in the Netherlands and consecutive investigations, the method appeared to be very suitable to assess the abundance of most invertebrate taxa in drinking water distribution systems and to be practicable for relatively inexperienced sampling and lab technicians. Although the numbers of small, less abundant or sessile taxa were not accurately assessed using the method, these taxa probably should not be the primary focus of monitoring by water utilities, as consumer complaints are not likely to be caused by these invertebrates. The accuracy of quantifying small invertebrates was further improved, however, by filtering the 100microm filtrate with a 30microm mesh plankton gauze filter.     

Dynamic diffusion model for tracing the real-time potential response of polymeric membrane ion-selective electrodes

A numerical solution for the prediction of the time-dependent potential response of a polymeric-based ion-selective electrode (ISE) is presented. The model addresses short- and middle-term potential drifts that are dependent on changes in concentration gradients in the aqueous sample and organic membrane phase. This work has important implications for the understanding of the real-time response behavior of potentiometric sensors with low detection limits and with nonclassical super-Nernstian response slopes. As a model system, the initial exposure of membranes containing the well-examined silver ionophore O,O' '-bis[2-(methylthio)ethyl]-tert-butylcalix[4]arene was monitored, and the large observed potential drifts were compared to theoretical predictions. The model is based on an approximate solution of the diffusion equation for both aqueous and organic diffusion layers using a numerical scheme (finite difference in time and finite elements in space). The model may be evaluated on the basis of experimentally available parameters and gives time-dependent information previously inaccessible with a simpler steady-state diffusion model. For the cases studied, the model gave a very good correlation with experimental data, albeit with lower than expected diffusion coefficients for the organic phase. This model may address numerous open questions regarding the response time and memory effects of low-detection-limit ion-selective electrodes and for other membrane electrodes where ion fluxes are relevant.     

Elimination of dimer formation in InIIIporphyrin-based anion-selective membranes by covalent attachment of the ionophore

The spontaneous hydroxy-bridged dimer formation of metalloporphyrins in ion-selective membranes gives rise to a short sensor lifetime (typically days), triggered by solubility problems, the occurrence of a super-Nernstian response slope, and a pH cross response. This dimer formation is eliminated here by covalent attachment of the ionophore to the polymer matrix. Specifically, two different indium(III)porphyrins containing polymerizable groups, the chloride-selective chloro(3-[18-(3-acryloyloxypropyl)-7,12-bis(1-methoxyethyl)-3,8,13,17-tetramethylporphyrin-2-yl]propyl ester)indium(III) and the nitrite-selective Chloro(5-(4-acryloyloxyphenyl)-10,15,20-triphenylporphyrinato)indium(III), were synthesized and copolymerized with methyl methacrylate and decyl methacrylate. The covalent attachment of the ionophore to the polymer matrix indeed prevents the metalloporphyrin from forming dimeric species, as confirmed by UV/visible spectroscopy. The ion-selective membranes with grafted indium porphyrin showed Nernstian response slopes to chloride, nitrite, perchlorate, and thiocyanate anions, with a selectivity comparable to membranes with freely dissolved or underivatized metalloporphyrin. The membranes containing grafted ionophores showed a lifetime of at least two months, apparently since crystallization of the poorly soluble dimeric species may no longer occur. This is one of the first examples where the covalent attachment of an ionophore drastically improves on a number of important sensor characteristics.