[Electrolyte balance in the nephrotic syndrome with cation exchange resin and ACTH administration].
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A combined precipitation-ion-exchange chromatographic method is described for the production of immunoglobulin G (IgG) from human plasma. The crude IgG fraction was precipitated with polyethylene glycol (PEG) and further purified on CM-Sephadex C-50. PEG and the contaminating proteins were not bound to the ion-exchange resin; they were eluted with the starting buffer. The purified IgG fraction was obtained in the second step by elution of the proteins bound to the column. In addition to column chromatography a batch procedure has been developed that requires relatively simple equipment. IgG of high purity was obtained by both column chromatography and batch procedure.
A method has been devised for the quantitative separation of trace amounts of alditols from mixtures with large amounts of monosaccharides, using a strongly basic ion-exchange resin. Ten kinds of common reducing monosaccharides used were strongly retained by a very basic anion-exchange resin in the hydroxyl form, whereas the corresponding alditols showed no significant affinity for the basic resin. Model studies showed excellent recoveries of alditol from known mixtures of alditol and the corresponding aldose at ratios in the range from 1 : 1X10(3) TO 1 : 1X10(4) (by weight). Application of this procedure to dextrans after reduction and hydrolysis resulted in quantitative separation of the terminal alditol.
A chromatin fraction enriched for Xenopus 5S RNA genes has been isolated by restriction endonuclease digestion and sucrose gradient velocity sedimentation. Soluble chromatin sedimenting at 70-80S contains approximately 50% of the oocyte-expressed 5S RNA genes and only 1.5-3% of total chromatin DNA; this represents a 15- to 30-fold purification of the 5S genes. Such chromatin isolated from somatic cells (blood and cultured kidney cells) retains the transcriptionally-inactive state of the oocyte-expressed 5S genes. Soluble chromatin from somatic cells prepared by micrococcal nuclease digestion also retains the inactive state of the oocyte-type 5S genes. It is likely that the level of chromatin structure responsible for inactivity of the oocyte genes in somatic cells is the nucleosome or short chains of nucleosomes and not supranucleosomal structures. The oocyte-type genes can be rendered transcriptionally active in somatic cell chromatin either by salt extraction of some chromosomal proteins or by treatment with the ion exchange resin Dowex A50W-X2. Alternatively, activation of these genes can be achieved by incubating somatic cell chromatin or nuclei with an extract prepared from Xenopus oocytes. This effect is not specific for 5S RNA genes as the transcription of other small RNAs (including pre-tRNA) is stimulated by the oocyte extract. The activating factor(s) is resistant to micrococcal nuclease, nondialyzable, heat labile and sensitive to trypsin; thus it is highly likely to be a protein or a group of proteins. Partial purification of the activating factor(s) has been achieved by ion exchange chromatography.
The concentration of the amino acids has been determined in granulocytes from healthy men and women by chromatographic analysis on ion-exchanging resins. Except taurine, ornithine and aspartic acid, the concentrations of all the amino acids are greater in the cells isolated from women. The most variable amino acids are alanine, valine, metionine, isoleucine, leucine, tyrosine, phenylalanine, lysine and histidine. Taurine represents 50% of the pool determined from men, whereas for women it represents only 25%.
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A method is presented for the preparation and assay of lactate dehydrogenase isoenzymes 1 and 2 as pure fractions. The total method involves the use of heat treatment to destroy the heat-labile fractions (isoenzymes 3, 4 and 5); fractionation using an ion-exchange resin; and immunochemical blocking of the muscle subunit, to remove any contamination of the fraction containing isoenzyme 1 with isoenzyme 2 and vice versa. Finally, the enzyme activities in the various fractions are measured using a standard kinetic assay procedure. Estimates can be obtained of all five isoenzymes if the heat treatment step is omitted.
The results of a series of pulse experiments carried out in a fixed bed column packed with a strongly acid ion-exchange resin and with reference to the separation of an aqueous sucrose-betaine mixture are presented. These data were used to evaluate equilibrium and diffusion parameters for both key components in the frame of linear chromatography theory. The estimated values were used to design a simulated moving bed plant aimed to separate betaine from sucrose in beet molasses.
The field dependence of the longitudinal and transverse nuclear magnetic relaxation rates of 23Na+ in aqueous crosslinked Na-poly(styrene sulfonate) (PSS) systems (ion exchange resins) has been obtained as a function of the degree of crosslinking. The relaxation is considerably enhanced relative to solutions of non-crosslinked NaPSS at equal ionizable group concentration. This is due to the dynamic constraints of the polymer chains, which render the averaging of the counter-ion chain interaction less efficient. The field dependence of the relaxation rates in the crosslinked NaPSS systems reveals two processes that are out of the extreme narrowing limit. This is in contrast to the relaxation behavior found in non-crosslinked NaPSS systems. To characterize these processes their correlation times were combined with constants of selfdiffusion to estimate the distances diffused by an ion in order to average the electric field gradient at its nucleus. These two distances are interpreted as characteristic length scales in the network. At all degrees of crosslinking it was found that the smallest of these length scales is roughly equal to the distance between two neighbouring crosslinks. The largest characteristic distance extends over several crosslinks and reflects inhomogeneities in the crosslink concentration. These conclusions were also reached from similar experiments on 7Li+ in LiPSS systems.
Phenolic compounds derived from the amino acid tyrosine are endogenous toxins, which are believed to be involved in the pathogenesis of hepatic coma. There are also xenobiotic phenolic substances, such as p-hydroxy-acetanilide (paracetamol or acetaminophen), which can lead to serious complications if taken in an overdose. In both cases, a drastic therapeutic measure such as haemoperfusion may be indicated to eliminate the toxin from the blood. In the present work, human serum has been dosed with the phenolic compounds of immediate relevance in exogenous and endogenous intoxication, and the effectiveness of various adsorbent materials for the elimination of the toxins from the serum has been investigated. Resins and charcoal in the native state have been compared with those encapsulated into large agarose beads, a process which improves the haemocompatibility and thus the practicability of the adsorbents. A certain degree of specificity has been observed. Whereas phenolic acids are adsorbed quite effectively onto the strongly basic ion exchange resins of the Dowex 1X type, particularly 1X8 or 2X8, phenol or paracetamol are less effectively eliminated. In contrast to many other classes of toxin, the Amberlite XAD-type resins are ineffective for all the phenolic substances investigated. Charcoal is the most effective adsorber in most cases, particularly when encapsulated in powder form into agarose beads.
A method is developed for determining the AMP-aminohydrolase activity by means of chromatography on the home KY-2 ion-exchange resin. It differs from the method of differential spectrophotometry by the fact that its application enables the content of IMP formed due to the enzymic reaction to be determined by its absolute quantity. This method yields satisfactory results independently of the substrate concentration. However, it cannot be applied for determination of the AMP-aminohydrolase activity in the presence of ATP.
A method is described for the purification of 3,3-dihydroxyazetidine (DHA), which accelerates the growth of Bifidobacterium spp., from the culture medium of Bacillus mesentericus (BM). Purification involved adsorption to an ion-exchange resin; it required less time and gave a higher yield than a previously reported method. Monitoring the inhibition of E. coli NIHJ JC-2, we searched for other strains that produced 3,3-dihydroxyazetidine. We found that not only B. mesentericus TO-A but also B. subtilis IFO13719 produced the compound of interest.
Several inborn errors of metabolism with abnormal polyol concentrations in body fluids are known to date. Most of these defects can be diagnosed by the assessment of urinary concentrations of polyols. We present two methods using tandem mass spectrometry for screening for inborn errors affecting polyol metabolism. Urine samples supplemented with internal standards ([13C4]erythritol, [13C2]arabitol and [2H3]sorbitol) were desalted by a mixed-bed ion-exchange resin. Separation was achieved by two different columns. Sugar isomers could not be separated using a Prevail Carbohydrate ES 54 column (method 1), whereas with the other column (Aminex HPX-87C) separation of the isomers was achieved (method 2). Multiple reaction monitoring polyol detection was achieved by tandem mass spectrometry with an electron ion-spray source operating in the negative mode. Age-related reference ranges of polyols (erythritol, treitol, arabitol, ribitol, xylitol, galactitol, mannitol, sorbitol, sedoheptitol and perseitol) in urine were established. The applicability of the method was demonstrated by the abnormal polyol concentrations observed in patients with transaldolase deficiency, ribose-5-phosphate isomerase deficiency and classical galactosaemia. This paper describes two methods for the analysis of urinary polyols by liquid chromatography-tandem mass spectrometry. Method 1 is a fast screening method with the quantification of total isomers and method 2 is a more selective method with the separate quantification of the polyols. Both methods can be used for diagnosing inborn errors of metabolism affecting polyol metabolism.
To develop an efficient method for the production of l-citrulline, optimum conditions for the conversion of l-arginine to l-citrulline by microbial l-arginine deiminase and for production of the enzyme were studied. A number of micro-organisms were screened to test their ability to form and accumulate l-citrulline from l-arginine. Pseudomonas putida was selected as the best organism. With this organism, enzyme activity as high as 9.20 units per ml could be produced by a shaking culture at 30 C in a medium containing glucose, ammonium phosphate, l-arginine hydrochloride, yeast extract, peptone, and inorganic salts. Appropriate addition of a surface active agent to the reaction mixture was found to shorten the time required for the conversion. A large amount of l-arginine hydrochloride was converted stoichiometrically to l-citrulline in 62 hr at 37 C. Accumulated l-citrulline was readily isolated in pure form by ordinary procedures with ion-exchange resins. Yields of isolated l-citrulline of over 90.5% from l-arginine hydrochloride were easily attainable.
The phenolic constituents of male and female flowers of Hagenia abyssinica were examined using chromatographic and spectroscopic techniques. Chromatographic analysis of kosins (phloroglucinols) was carried out using TLC, reversed-phase LC and glass-capillary GLC. Purification of the crude kosins was effected by silica gel column chromatography and purification of phenolic acids using ion-exchange resins and preparative TLC on silica gel. The kosins were characterized by TLC, LC, UV, IR, MS and H1 NMR spectroscopy while the phenolic acids were characterized by UV-spectroscopy, TLC and LC. The phenolic acids found were protocatechuic acid, p-hydroxybenzoic acid and vanillic acid. The content of the kosins and phenolic acids was established quantitatively for both male and female flowers.
The acidic glycosaminoglycans (AGAG) in normal portions of human gastric tissue were separated by electrophoresis in 3 buffer systems. Paper chromatographic separation of the constitutional disaccharide units by digestion of chondroitin sulfates (CS) with chondroitinase-ABC and chondroitinase-AC was carried out after fractionation of CS by ion-exchange resin column chromatography. Thin-layer chromatography of hexosamines and other biochemical analysis were also performed. The presence of hyaluronic acid in the gastric tissue was substantiated by the enzymatic susceptibility to streptomyces hyaluronidase. The results indicated that human gastric AGAG consisted of, in the order of amount, heparan sulfates, dermatan sulfate, hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate and presumably oversulfated chondroitin sulfate.
A thin-layer chromatographic method for the detection and determination of Paraquat in biological materials is presented. Paraquat was concentrated efficiently with the ion-exchange resin Dowex 50W-4 using a batch operation. Of the 16 solvent systems used the best results were obtained on silica gel G plates with methanol : 6N hydrochloric acid (20 : 30) or methanol : 6N sulfuric acid (20 : 30); and on cellulose plates with methanol : 6N hydrochloric acid (35 : 15) or methanol : 6N sulfuric acid (40 : 10). Also described is a microplate method for the thin-layer chromatography of Paraquat.
In the semiconductor industry, there is interest in determining borate at sub-ppb levels in ultrapure water, since borate is an early breakthrough ion from ion-exchange resin beds. Although dissolved silica is the most common species currently used to monitor the breakdown of the deionization systems, it is thought that borate probably breaks through earlier than silicate. To be of use as an early-warning indicator, borate must be determined at ppt levels. This paper discusses benchtop results with several new column products designed to deliver low-ppt detection limits for boron as borate. The system uses a prototype borate-specific concentrator column that is coupled to an ion-exclusion separator and suppressed-conductivity detection. The acidic eluent, containing mannitol, quantitatively elutes the borate from the concentrator. The analytical separation is performed using a specially designed ion-exclusion column. Data presented are from two multilevel calibration studies. Included is a discussion of detection-limit calculations and recommended formats for reporting results.