20-400 Sigma-AldrichMagna GrIP™ ラック(8ウェル)

The factors affecting micellar electrokinetic capillary chromatographic separation of hippuric and o-, m-, p-methylhippuric acid were investigated by changing the species of micelles, and adding urea to the micellar solution. The analysis of hippurates in human urine is demonstrated under optimum conditions using 20 mM phosphate buffer (pH 8.0) containing 100 mM dodecyltrimethylammonium bromide and 4 M urea at -22 kV applied voltage. This method proved suitable for the screening of hippurates in human urine following occupational exposure to toluene and xylene.

A novel iron-sulfur protein has been isolated from the sulfate-reducing bacterium Desulfovibrio vulgaris (Hildenborough). It is a stable monomeric protein, which has a molecular mass of 52 kDa, as determined by sedimentation-equilibrium centrifugation. Analysis of the metal and acid-labile sulfur content of the protein revealed the presence of 6.3 +/- 0.4 Fe/polypeptide and 6.2 +/- 0.7 S2-/polypeptide. Non-iron transition metals, heme, flavin and selenium were absent. Combining these data with the observation of a very anisotropic S = 1/2 [6Fe-6S]3+ prismane-like EPR signal in the dithionite-reduced protein, we believe that we have encountered the first example of a prismane-cluster-containing protein. The prismane protein has a slightly acidic amino acid composition and isoelectric point (pI = 4.9). The ultraviolet/visible spectrum is relatively featureless (epsilon 280 = 81 mM-1.cm-1, epsilon 400 = 25 mM-1.cm-1, epsilon 400,red = 14 mM-1.cm-1). The shape of the protein is approximately globular (S20.w = 4.18 S). The N-terminal amino acid sequence is MFS/CFQS/C QETAKNTG. Polyclonal antibodies against the protein were raised. Cytoplasmic localization was inferred from subcellular fractionation studies. Cross-reactivity of antibodies against this protein indicated the occurrence of a similar protein in D. vulgaris (Monticello) and Desulfovibrio desulfuricans (ATCC 27774). We have not yet identified a physiological function for the prismane protein despite trials for some relevant enzyme activities.

An HPLC method for the analysis of levamisole and thiabendazole has been developed with recoveries varying over 63-75%. The two anthelmintics are extracted from meat with ethyl acetate, purified by liquid/liquid extraction and analyzed quantitatively on a mu Bondapak C18 column. The optimum detection is achieved by means of a photodiode array detector at 240 nm for levamisole and at 300 nm for thiabendazole. The detection limits for both compounds in meat are 25 micrograms/kg and 5 micrograms/kg, respectively.

The effects of n-3 fatty acids on plasma lipids, lipoproteins and apoproteins have usually been studied in humans after feeding of purified fish oil. This study describes the effect of a natural diet, containing salmon as the source of n-3 fatty acids, on these parameters as compared to a diet very low in n-3 fatty acids. The subjects were nine normolipidemic, healthy males who were confined to a nutrition suite for 100 days. During the first 20 days of the study the participants were given a stabilization diet consisting of 55% carbohydrates, 15% protein, and 30% fat. The n-3 content of this diet was less than 1%, and it contained no 20- or 22-carbon n-3 fatty acids. After the stabilization period the men were split into two groups, one group continued on the stabilization diet while the other received the salmon diet that contained approximately 2.1 energy percent (En%) of calories from 20- and 22-carbon n-3 fatty acids. Both diets contained equal amounts of n-6 fatty acids. This regime continued for 40 days, then the two groups switched diets for the remainder of the study. Plasma triglycerides were lowered significantly (p less than 0.01) and high density lipoprotein cholesterol (HDL-C) was significantly elevated (p less than 0.01) after the men consumed the salmon diet for 40 days. The very low density lipoproteins (VLDL) were lowered, but the trend did not reach statistical significance during the intervention period.(ABSTRACT TRUNCATED AT 250 WORDS)

We have measured the fluctuations in the current through gramicidin A (GA) channels in symmetrical solutions of monovalent cations of various concentrations, and compared the spectral density values with those computed using E. Frehland's theory for noise in discrete transport systems (Frehland, E. 1978. Biophys. Chem. 8:255-265). The noise for the transport of NH4+ and Na+ ions in glycerol-monooleate/squalene membranes could be accounted for entirely by shot noise in the process of transport through a single-filing pore with two ion binding sites. However, in confirmation of results in a previous paper (Sigworth, F. J., D. W. Urry, and K. U. Prasad. 1987. Biophys. J. 52:1055-1064) currents of Cs+ showed a substantial excess noise at low ion concentrations, as did currents of K+ and Rb+. The excess noise was increased in thicker membranes. The observations are accounted for by a theory that postulates fluctuations of the entry rates of ions into the channel on a time scale of approximately 1 microsecond. These fluctuations occur preferentially when the channel is empty; the presence of bound ions stabilizes the high conductance conformation of the channel. The fluctuations are sensed to different degrees by the various ion species, and their kinetics depend on membrane thickness.

The primary structure of rabbit J chain, which occurs covalently bound to secretory IgA, was determined. J chain was isolated in its S-carboxymethylated form, in one step, by SDS/PAGE followed by electro-elution; 5 nmol of protein (approx. 75 micrograms), in all, was necessary for the determination of the complete sequence by the 'shot-gun' microsquencing technique; with the use of several site-specific endoproteinases, the various digests of S-carboxymethylated J chain were separated by micro-bore reverse-phase h.p.l.c. and the partial N-terminal sequences of all peptides were analysed. From the sequence alignment, gaps were filled by further extensive sequencing of the relevant overlapping fragments isolated from selected digests. Rabbit J chain comprises 136 amino acid residues, out of which eight are conserved cysteine residues, and is more closely similar to the human sequence (73.5% identify) than to the mouse sequence (68% identity). There is one unique glycosylation site at asparagine-48.