Choline measurement

Phosphatidylcholine is an important component of cell membranes but cell mem branes are more than simply lipid bilayers Although their composition varies with their source a typical membrane contains about equal amounts of lipid and protein and the amount of cholesterol m the lipid fraction can approximate that of phosphatidylcholine The lipid fraction is responsible for the structure of the membrane Phosphatidyl choline provides the bilayer that is the barrier between what is inside the cell and what IS outside Cholesterol intermingles with the phosphatidylcholine to confer an extra measure of rigidity to the membrane... 

Johnson has described a method for the estimation of physostigmine in the salicylate for the determination of minute amounts of the alkaloid Ellis, Plaehte and Straus have devised processes depending on (a) inhibition of serum choline-esterase by the alkaloid, or (b) measurement of the colour intensity produced by the conversion of physostigmine to rubreserine in an alkaline medium. i ... 

There is also inside-outside (transverse) asymmetry of the phospholipids. The choline-containing phospholipids (phosphatidylcholine and sphingomyelin) are located mainly in the outer molecular layer the aminophospholipids (phosphatidylserine and phos-phatidylethanolamine) are preferentially located in the inner leaflet. Obviously, if this asymmetry is to exist at all, there must be limited transverse mobility (flip-flop) of the membrane phospholipids. In fact, phospholipids in synthetic bilayers exhibit an extraordinarily slow rate of flip-flop the half-life of the asymmetry can be measured in several weeks. However, when certain membrane proteins such as the erythrocyte protein gly-cophorin are inserted artificially into synthetic bilayers, the frequency of phospholipid flip-flop may increase as much as 100-fold. 

ACh is widely distributed throughout the brain and parts of the spinal cord (ventral horn and dorsal columns). Whole brain concentrations of lOnmolg" tissue have been reported with highest concentrations in the interpeduncular, caudate and dorsal raphe nuclei. Turnover figures of 0.15-2.0 nmol g min vary with the area studied and the method of measurement, e.g. synthesis of labelled ACh from [ " C]-choline uptake or rundown of ACh after inhibition of choline uptake by hemicholinium. They are all sufficiently high, however, to suggest that in the absence of synthesis depletion could occur within minutes. 

Heig, M.E., Ulrich, D., Sagelsdorff, P., Richter, C. and Lutz, W.K. (1990). No measurable increase in thymidine glycol or 8-hydroxydeoxyguanosine in liver DNA of rats treated with nafenopin or choline-devoid low-methionine diet. Mut. Res. 238, 325-329. 

A flow injection optical fibre biosensor for choline was also developed55. Choline oxidase (ChOX) was immobilized by physical entrapment in a photo-cross-linkable poly(vinyl alcohol) polymer (PVA-SbQ) after adsorption on weak anion-exchanger beads (DEAE-Sepharose). In this way, the sensing layer was directly created at the surface of the working glassy carbon electrode. The optimization of the reaction conditions and of the physicochemical parameters influencing the FIA biosensor response allows the measurement of choline concentration with a detection limit of 10 pmol. The DEAE-based system also exhibited a good operational stability since 160 repeated measurements of 3 nmol of choline could be performed with a variation coefficient of 4.5%. 

With the specific suitable oxidases, lactate, choline and glucose could be assayed. Concentration measurements of these metabolites could be performed over at least two decades wit a detection limit of 10 pM for lactate and choline and 20 pM for glucose. 

Although the stabilizing interactions between the amino acid side chains of PLC/j, and the choline headgroup are readily apparent in the PLC fc-phosphonate inhibitor complex, it is more difficult to identify contacts between the protein and the acyl chains of the inhibitor [45]. In part this is because thermal motion in the acyl side chains, especially the sn-1 chain, renders them somewhat disordered. Consequently, the measured distances between the side chain carbons... 

Sanders The esterase destroys acetylcholine as an effective agonist, so you can still see the consequences of acetylcholine release by measuring tritiated choline, but it is not an effective agonist. 

Figure 16 shows the experimental arrangement for the measurement of the surface pressure. The trough (200 mm long, 50 mm wide and 10 mm deep) was coated with Teflon. The subphase temperature was controlled within 0.1 C by means of a jacket connected to a thermostated water circulator, and the environmental air temperature was kept at 18 °C. The surface tension was measured with a Wilhelmy plate of platinum(24.5 x 10.0 x 0.15 mm). The surface pressure monitored by an electronic balance was successively stored in a micro- computer, and then Fourier transformed to a frequency domain. The surface area was changed successively in a sinusoidal manner, between 37.5 A2/molecule and 62.5 A2/molecule. We have chosen an unsaturated phospholipid(l,2-dioleoyl-3-sn-phosphatidyI-choline DOPC) as a curious sample to measure the dynamic surface tension with this novel instrument, as the unsaturated lipids play an important role in biomembranes and, moreover, such a "fluid" lipid was expected to exhibit marked dynamic, nonlinear characteristics. The spreading solution was 0.133 mM chloroform solution of DOPC. The chloroform was purified with three consecutive distillations. 

The pA a shift can be directly measured by the solvatochromic shift of the ultraviolet absorption spectra. For PCP, the p%lir is 5.97 in phosphatidyl choline membranes, and increases up to 6.78 in the negatively charged phosphatidyl glycerol membranes [123], The addition of cholesterol decreases the pATam again slightly in both types of membranes. 

Fig. 15. Comparison of a water suppressed muscle spectrum and a spectrum from yellow bone marrow containing almost pure fat (triglycerides). Measurement parameters STEAM sequence, TE=10 ms, TM=15 ms, TR = 2 s, 40 acq., VOI (11 X 11 X 20) mm. (a) Spectrum from TA muscle recorded after careful positioning of the VOI, avoiding inclusion of macroscopic fatty septa allows separation of extramyocellular (EMCL, broken lines) and intramyocellular lipid signals (IMCL, dotted lines) based on susceptibility differences. For this reason characteristic signals from fatty acids occur double. Signals of creatine (methyl, Crs, and methylene, Cr2) show triplet and doublet structure, respectively, due to dipolar coupling effects. Further signals of TMA (including carnitine and choline compartments), Taurine (Tau), esters, unsaturated fatty acids (-HC=CH-), and residual water are indicated, (b) Spectrum from yellow fatty bone marrow of the tibia with identical measuring parameters, but different amplitude scale.

They did not report the minimum lethal dose or any other quantitative measure of toxicity, but nobody reading their paper would have missed the fact that the compounds are violently poisonous. During World War II, research on the fluorophosphonates was carried on for military purposes, and Adrian and his coworkers in Britain noted the similarity between the physiological action of the fluorophosphonates and that of reversible inhibitors of choline esterase (Adrian et al., 1947). This led to a number of scientific investigations of the action of nerve gases on various esterases. 

Fig. 11. Evidence that a membrane-associated immunochemical reaction (complement fixation) depends on the mobility of the target hapten (IX) in the plane of a model membrane. The extent of the immunochemical reaction, complement fixation, is measured by A Absorbance at 413 nm. Temperature is always 32°C, which is above the chainmelting temperature (23°C) of dimyristoylphosphatidylcholine used for the data given in A and below the chain-melting transition temperature (42°C) of dipalmitoylphosphatidyl-choline used for the data in B. Thus A refers to a fluid membrane and B refers to a solid membrane. The numbers by each curve are equal to c, the mole % of spin-label hapten IX in the plane of the lipid membrane. It will be seen that complement fixation, as measured by A Absorbance at 413 nm is far more effective in the fluid membrane than in the solid membrane at low hapten concentrations (i.e., c 0.3 mo e%). In C the lipid membrane host is a 50 50 mole ratio mixture of cholesterol and dipalmitoylphosphatidylcholine. The immunochemical data suggest that this membrane is in a state of intermediate fluidity. Specific affinity-purified IgG molecules were used in these experiments. (For further details, see Ref. 5.)... 

Measurement of Acetylcholine Levels by pH Changes The concentration of acetylcholine (a neuro-transmitter) in a sample can be determined from the pH changes that accompany its hydrolysis. When the sample is incubated with the enzyme acetylcholinesterase, acetylcholine is quantitatively converted into choline and acetic acid, which dissociates to yield acetate and a hydrogen ion ... 

Alkyl and alkenyl ethers have been identified in phosphatidyl choline and phosphatidyl ethanolamine fractions recovered from whole milk (Hay and Morrison 1971). To our knowledge, levels of alkyl and alkenyl ethers in phospholipids of milk lipid globule membrane have not been measured. 

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