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1.
D. T. Edmonds 《European biophysics journal : EBJ》1987,14(4):195-201
Most current models of membrane ion channel gating are abstract compartmental models consisting of many undefined states connected by rate constants arbitrarily assigned to fit the known kinetics. In this paper is described a model with states that are defined in terms of physically plausible real systems which is capable of describing accurately most of the static and dynamic properties measured for the sodium channel of the squid axon. The model has two components. The Q-system consists of charges and dipoles that can move in response to an electric field applied across the membrane. It would contain and may compose the gating charge that is known to transfer prior to channel opening. The N-system consists of a charged group or dipole that is constrained to move only in the plane of the membrane and thus does not interact directly with the trans-membrane electric field but can interact electrostatically with the Q-system. The N-system has only two states, its resting state (channel closed) and its excited state (channel open) and its response time is very short in comparison with that of the Q-system. On depolarizing the membrane the the N-system will not make a transition to its open state until a critical amount of Q-charge transfer has occurred. Using only four adjustable parameters that are fully determined by fitting the equilibrium properties of the model to those of the sodium channel in the squid axon, the model is then able to describe with some accuracy the kinetics of channel opening and closing and includes the Cole and Moore delay. In addition to these predictions of the behaviour of assemblies of channels the model predicts some of the individual channel properties measured by patch clamp techniques. 相似文献
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3.
Results from this laboratory have demonstrated that14C-labeled myelin opsonized with antibodies raised to purified CNS myelin in rabbit is phagocytized by cultured macrophages in larger amounts than untreated myelin or myelin opsonized with preimmune serum. The cultured macrophages produced high amounts of radioactive cholesterol ester and triglyceride from the antibody-treated myelin while much less was formed from preimmune serum-treated or untreated myelin. Antiserum to galactocerebroside also greatly enhanced the formation of radioactive cholesterol ester, while that to myelin basic protein as well as to other myelin constituents had little or no effect. Serum from Lewis rats with acute EAE 13–14 days after immunization with whole CNS myelin also stimulated radioactive cholesterol ester formation compared to serum from Freund's adjuvant-injected controls (FAC). Serum from EAE rats as a result of myelin basic protein injection was as active as that from rats with whole myelin injection. No galactocerebroside antibody could be demonstrated in the EAE sera, although a strong immunostaining to myelin basic protein and proteolipid protein was demonstrated. IgG prepared from EAE serum also showed stimulatory effects compared to IgG from FAC serum, but much of the activity was lost, and the possibility that other factors may be involved is discussed. These experiments provide evidence that myelin phagocytosis and digestion by macrophages is enhanced by the presence of antibody to myelin. In EAE this antibody may leak into CNS with the breakdown of the blood-brain barrier. A humoral involvement in demyelination in EAE is implicated, and these findings may be extended eventually to the demyelinative mechanism in multiple sclerosis where IgG is found in large amounts in the CNS.Special Issue Dedicated to Dr. Abel Lajtha. 相似文献
4.
Robust estimation of standard curves for protein molecular weight and linear-duplex DNA base-pair number after gel electrophoresis 总被引:13,自引:0,他引:13
An accurate procedure for estimating linear-duplex DNA base-pair numbers and protein molecular weights after electrophoresis in single concentration gels is presented. A robust modified hyperbola was found to be superior for determining molecular weights and base-pair numbers for a set of known standards when compared with the conventional log transformation and a similar hyperbolic model. We describe the use of a soft laser-scanning densitometer to measure band-migration distances of wet, stained polyacrylamide gels for proteins and photographic negatives of agarose gels containing DNA stained with ethidium bromide. This automated densitometric method was more accurate than existing methods. A BASIC computer program detailing the procedure is included. 相似文献
5.
Marion Edmonds Smith Florentino P. Somera Terry J. Sims 《Journal of neurochemistry》1985,45(4):1205-1212
The enzyme UDP-N-acetylglucosamine: dolichyl phosphate, N-acetylglucosamine-1-phosphate transferase initiates the synthesis of the oligosaccharide chain of complex-type glycoproteins. In view of the high content of glycoprotein in peripheral nerve myelin, the properties of this enzyme, its changes with age, and the effect of the specific inhibitor tunicamycin were investigated. The enzyme activity in rat peripheral nerve homogenate was completely dependent on the presence of exogenous dolichyl phosphate as well as Mg2+ and a detergent (Triton X-100) and was also greatly stimulated by a high salt concentration (0.4 M KCl) and AMP. The highest specific activity was present in the postmitochondrial membranes. The specific activity in postmitochondrial membranes in the presence of exogenous dolichyl phosphate reached a maximum at 17 days and remained relatively high throughout development, up to 2 years of age, but the activity was much lower when dolichyl phosphate was not added. This indicates that the enzyme level does not decrease with age, but that the content of the lipid cofactor may limit glycoprotein synthesis in vivo. Tunicamycin (5 micrograms) was injected intraneurally into 24-day-old rat sciatic nerve, and the enzyme was assayed from 1 to 24 days after injection. The specific activity of the transferase remained at low levels (5-40% of the level in control nerve) in most injected nerves assayed throughout this postinjection period. A protein previously identified as the unglycosylated P0 protein was synthesized in vitro by the tunicamycin-injected nerve and could be demonstrated to be incorporated into myelin in large amounts at 2 days and in small amounts at 6 days after injection.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
6.
D T Edmonds 《Proceedings of the Royal Society of London. Series B, Containing papers of a Biological character. Royal Society (Great Britain)》1986,228(1250):71-84
An alternative model is presented for an ionic counterport that depends upon electrostatic rather than steric forces. It consists of two passive ion channels, one selective for I-type ions and the other for J-type ions. The ions interact electrostatically such that the presence of one type of ion within its channel affects the motion of the second type of ion within its channel. In these circumstances it is possible to arrange that the spontaneous flow of I ions across the membrane, down their electrochemical potential gradient, pumps J ions in the opposite direction across the membrane, against their electrochemical gradient. To illustrate this type of model, a particular example of interionic coupling is described in which both types of ion interact with the electric dipole moments of some membrane-spanning alpha-helical sections of the counterport protein complex. By assuming that a group of four alpha-helices is free to rotate slightly about an axis perpendicular to the membrane, the desired form of coupling is obtained. Making simplifying assumptions, it is possible to calculate the kinetics of the model and to compare these with those expected in real counterports. Finally it is shown that, if the helix group rotation is powered by an external energy source, the pair of coupled passive ion channels can mimic a primary exchange pump such as Na+-K+ ATPase. Here both types of ion are propelled in opposite directions across the membrane and simultaneously against their electrochemical potential gradients. 相似文献
7.
A technique for differentiating high-resolution NMR signals from different regions of small objects is outlined and some initial results on model systems are given. This method uses inorganic paramagnetic or diamagnetic ions to create magnetic field gradients at phase boundaries. 相似文献
8.
Lipids of Pseudomonas aeruginosa Cells Grown on Hydrocarbons and on Trypticase Soy Broth 总被引:3,自引:1,他引:2
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Lipids were extracted from cells of Pseudomonas aeruginosa grown on a pure hydrocarbon (tridecane), mixed hydrocarbons (JP-4 jet fuel), and on Trypticase Soy Broth. Total lipids produced from each substrate represented from 7.1 to 8.2% of cellular dry weight, of which 5.0 to 6.4% were obtained before cellular hydrolysis (free lipids) and 1.7 to 2.0% were extracted after cellular hydrolysis (bound lipids). Free lipids from cells grown on each medium were separated into four fractions by thin-layer chromatography. All fractions were present in cells from each type of medium, and the "neutral fraction" constituted the largest fraction. The fatty acid composition of free lipids was determined by gas-liquid chromatography. Cells grown on each medium contained saturated and unsaturated C(14) to C(20) fatty acids. Trace amounts of C(13) fatty acids were found in tridecane-grown cells. Saturated C(16) and C(18) were the major acids present in all cells. Quantitative differences were found in fatty acids produced on the three media, but specific correlations between substrate carbon sources and fatty acid content of cells were not evident. Tridecane-grown cells contained only traces of C(13) acid and small amounts of C(15) and C(17) acids, suggesting that the organism's fatty acids were derived from de novo synthesis rather than by direct incorporation of the hydrocarbon. 相似文献
9.
Two fuel utilizers, a strain of Pseudomonas aeruginosa and a Hormodendrum sp., and two fuel isolates which did not use fuel, Staphylococcus and Bacillus spp., were tested for ability to survive and grow in systems containing emulsified or nonemulsified forms of JP-4 jet fuel. Neither emulsion (Alamac no. 1 or Alamac no. 2) supported microbial growth without a water phase. Growth of P. aeruginosa in liquid systems containing either emulsion was not significantly different from growth in liquid systems with nonemulsified fuel. The Hormodendrum sp. grew well in a liquid medium containing nonemulsified JP-4, but when either of the emulsions served as carbon source no growth was observed. However, good growth was noted on spread plates with either emulsion. Viable cells of Bacillus sp. did not increase over a 4-day period, and Staphylococcus sp. did not survive in liquid systems containing JP-4 or emulsions. 相似文献
10.
Seventy-two samples from jet aircraft fuel systems were examined for microbial contamination. Ten contaminated samples yielded 43 microorganisms which were classified into nine genera of bacteria and three genera of fungi. The predominant types, comprising about 37% of the isolated cultures, were identified as Bacillus spp. The remaining cultures were distributed among 11 genera, each of which represented 2 to 9% of the total isolates. Four cultures could not be assigned to a genus on the basis of the diagnostic criteria used. Only five isolates, in the genera Pseudomonas and Hormodendrum (Cladosporium), grew abundantly in a mineral salts solution with JP-4 fuel as the sole source of carbon. The presence of fuel utilizers in a fuel system may be a better index to potential problems that have been correlated with microbial contamination than the presence of aerobic sporeforming bacilli. 相似文献