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Query: UMLS:C0009443 (cold)
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1. A critical study was made of the quantitative extraction of nucleotide and sugar phosphates from plant tissue by either boiling aqueous ethanol or cold trichloroacetic acid. The effect of the extraction technique on the inactivation of the enzymes in the plant tissue and the possibility of adsorption of the phosphate esters on the cell wall were especially considered. 2. In the recommended method the plant tissue was frozen in liquid nitrogen, ground to a powder and then blended with cold aqueous trichloroacetic acid containing 8-hydroxyquinoline to prevent adsorption. 3. The extract contained large amounts of trichloroacetic acid, cations, chloride, sugars, amino acids, hydroxy organic acids, phytic acid, orthophosphoric acid and high-molecular-weight material including some phosphorus-containing compounds. All of these were removed as they were liable to interfere with the chromatographic or enzymic assay of the individual nucleotide or sugar phosphates. 4. The procedure was as follows: the last traces of trichloroacetic acid were extracted with ether after the solution had been passed through a column of Dowex AG 50 in the hydrogen form to remove all cations. High-molecular-weight compounds were removed by ultrafiltration and low-molecular-weight solutes by a two-stage chromatography on cellulose columns with organic solvents. In the first stage, sugars, amino acids, chloride and phytic acid were separated by using a basic solvent (propan-1-ol-water-aqueous ammonia) and, in the second stage, the organic acids and orthophosphoric acid were separated by using an acidic solvent (di-isopropyl ether-formic acid-2-methylpropan-2-ol-water). The final solution of nucleotide and sugar phosphates was substantially free from other solutes and was suitable for the detection of individual phosphate esters by either chromatography or enzymic assay. 5. The recovery of d-glucose 6-phosphate or adenosine 5'-triphosphate added to a trichloroacetic acid extract simulating that from peas and potatoes, and isolated according to the standard procedures, was better than 95%. Estimation of naturally occurring d-glucose 6-phosphate and adenosine 5'-triphosphate in the initial extract of peas and potatoes and in the final purified extract also indicated a recovery of about 95%. A similar estimation of uridine diphosphate glucose in potatoes showed that little or no breakdown occurred.
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PMID:Analysis of phosphate esters in plant material. Extraction and purification. 604 32

1. A study was made of the composition and structure of walls isolated from yeast grown in continuous culture at different rates, under three conditions of glucose limitation in which the concentrations of glucose and ammonium sulphate in the medium and the oxygen-transfer rate in the culture were varied, and one condition of NH(4) (+) limitation. 2. The contents of total glucan and total mannan in the walls were relatively little affected by the growth rate under any of the four sets of conditions. The phosphorus and protein contents of walls from yeast grown under each of the four conditions increased as the growth rate was decreased. Walls from yeast grown under NH(4) (+) limitation contained only half as much protein as walls from cells grown under glucose limitation. The proportion of lipid was greatest in walls from yeast grown under NH(4) (+) limitation. 3. A procedure was devised for fractionating isolated walls, based on the ease with which the glucan and mannan were extracted with water and with hot and cold 6% (w/v) potassium hydroxide solution. The percentage of glucan, mannan, protein and phosphorus in each of the fractions was affected by the rate of growth and by the nature of the substrate limitation. 4. The beta-fructofuranosidase activities of yeast grown under glucose limitation increased as the growth rate was lowered, but decreased at very low growth rates. The effects at low growth rates were probably due to repression of enzyme synthesis by residual glucose in the culture filtrate. The beta-fructofuranosidase activities of yeast grown under NH(4) (+) limitation were much lower than those from yeast grown under any of the conditions of glucose limitation. 5. Yeast cells grown at any of the rates under NH(4) (+) limitation were longer and thinner than those grown at the same rate under any of the conditions of glucose limitation. Mean cell volumes were dependent on growth rate but not on the nature of the substrate limitation. 6. Electron micrographs of thin sections of isolated walls showed that cells grown under NH(4) (+) limitation had a more porous structure than those from cells grown under any of the conditions of glucose limitation.
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PMID:Effect of growth rate and substrate limitation on the composition and structure of the cell wall of Saccharomyces cerevisiae. 605 21

The composition of normal, hypertrophied, or sodium-loaded rabbit portal anterior mesenteric vein and of normal and sodium-loaded guinea pig taenia coli smooth muscle was measured in cryosections with electron probe analysis, and the effects of wash with cold sodium-free (Lithium) solutions were determined. There was no significant difference in the cytoplasmic, nuclear, or mitochondrial concentrations of any of the measured elements (sodium, potassium, chloride, magnesium, calcium, phosphorus, sulfur) between hypertrophied, sham-operated, or control veins. The cytoplasmic potassium:sodium:chloride ratio in rabbit portal anterior mesenteric vein was 1:0.26:0.46, and the average sodium concentration (198 mmol/kg dry cytoplasmic weight) was nearly twice as high as estimated from ion flux measurements. The cytoplasmic sodium concentration of normal guinea pig taenia coli was 61 mmol/kg dry weight. The existence of a rapidly exchanging, relatively low affinity, and temperature-insensitive component of cytoplasmic sodium efflux was indicated by the reduction in cytoplasmic sodium after washout in cold, sodium-free (lithium or Tris-substituted) solutions. This reduction, by 62% in normal, 71% in sodium-loaded portal anterior mesenteric vein, and 36% in sodium-loaded guinea pig taenia coli smooth muscle, suggests that the lithium wash method can underestimate cell sodium. In sodium-loaded guinea pig taenia coli and portal anterior mesenteric vein smooth muscles, the cytoplasmic sodium analyzed in individual cells showed a bimodal distribution; in cryosections, the cells having the highest sodium and lowest potassium and phosphorus content also had a more electronlucent (light cell) appearance.
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PMID:Electron probe analysis of sodium and other elements in hypertrophied and sodium-loaded smooth muscle. 632 Oct 54

Two distinct carbohydrate antigens were isolated from the cell surface of group B streptococcus, type II. One antigen was extracted from SDS-purified cell walls by cold trichloroacetic acid and contained galactose, glucose, rhamnose, glucosamine and sialic acid in the approximate molar proportions 1.7:1.0:3.4:0.9:0.21 respectively. The serological activity of this polymer indicated that it is the group-specific antigen common to all group B streptococci. The second antigen was extracted by phenol from cell membranes and contained galactose, glucose, glucosamine, phosphorus and fatty acid in a molar ratio of 1.6:1.0:0.35:2.6:0.016 respectively. This antigen was shown to be specific for type II, group B streptococcus.
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PMID:The isolation and immunochemical characterisation of a cell-wall carbohydrate and a membrane lipocarbohydrate antigen of group B streptococcus, type II. 633 63

Serum chemical values were determined in cold-stressed Holstein bull calves ranging from 1 to 7 days of age. The animals were anesthetized and cold-stressed until their core body temperature (colonic) was lowered 10 C. Animals were then rewarmed in warm water, with heat pads or heat lamps, or were allowed to recover naturally (unassisted) at room temperature. Blood samples were collected at selected intervals during cooling and recovery. Increases (P less than 0.05) were observed in the concentrations of glucose, calcium, phosphorus, iron, alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, total protein, albumin, total globulin, serum urea nitrogen, uric acid, total bilirubin, indirect bilirubin, and cholesterol in the cold-stressed calves during cooling. Concentrations of chloride and insulin decreased (P less than 0.05) during the same period. Changes observed in many of the serum chemical values during rewarming were generally the reverse of the respective changes that occurred during cooling, although insulin values became exceedingly high in some cases midway or near the end of recovery. Serum enzyme values also remained high during most of recovery. Data did not indicate a clear advantage of one method of rewarming over the other methods used in terms of return of the serum chemical values to normal.
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PMID:Serum chemical values in hypothermic and rewarmed young calves. 634 64

Microbial requirements for P were assumed to be a function of the amount of microbial protein synthesis (microbial growth) and of the quantity of organic matter (OM) fermented in the rumen. The relationships among P incorporation into microbial matter and protein synthesis, ammonia utilization, volatile fatty acid (VFA) production and organic matter fermented (OMF) were studied in short-term incubations (3 h) using 32P-labelled phosphate. The amount of P incorporated was calculated from extracellular phosphate pool specific activity and the radioactivity incorporated into the microbial sediment during incubation (table 1). The inocula came from sheep fed a protein-free purified diet. In order to vary the intensity of fermentation, carbohydrates with a wide range of degrees of enzymatic susceptibility were used as substrates and the medium was either provided or was deficient in S and trace elements (table 4). Nitrogen was supplied as ammonium salts. Linear regression analyses showed that P incorporation was positively correlated with the criteria of protein synthesis and OM fermentation (figs. 1, 2, 3, 4). However, there was significant phosphorus incorporation when the value for nitrogen incorporation was zero (equation A: (Pi (mg) = 0.162 NH3-N + 0.376; r = 0.9). This was assumed to result either from energetic uncoupling (fermentation without concomitant bacterial growth) or from the lysis of cold microbial cells only. Equation A would reflect total P incorporation and equation A' Pi (mg) = 0.162 NH3-N (mg), net P incorporation. It was assumed that in vitro microbial requirements for P were in the range of 30-70 mg of P/liter of medium for 3-hour incubation, depending on the intensity of fermentation. From a mean value of microbial N yield of 30 g/kg of DOMR (organic matter apparently digested in the rumen), it was calculated that the total and net P requirements in vivo were 6 and 4.9 g/kg of DOMR, respectively, corresponding to 3.9 and 3.2 g/kg of DOM (digestible organic matter). From equation D, relating Pi to OMF, the P requirements were about 4.4 g/kg of DOM. It is suggested that microbial requirements for P varied from 3 to 5 g of P/kg of DOM, depending on the efficiency of microbial synthesis and the extent of carbohydrate fermentation. These results, considered as indicative, should be checked in in vivo experiments.
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PMID:[In vitro estimation using radioactive phosphorus of the phosphorus requirements of rumen microorganisms]. 635 Dec 7

A general procedure is described for the determination of acrylonitrile (AN) in foods such as margarine, honey butter, cold-pack cheese, and peanut butter, which are likely to be packaged in AN-based plastic. The entire sample is blended with water and salt at less than 5 degrees C, aliquots are sealed in crimp-top vials, and the vials are equilibrated in a boiling water bath. The headspace is sampled by using a heated syringe, and AN is determined by gas chromatography with a nitrogen-phosphorus selective detector. The inclusion of propionitrile as an internal standard allows quantitation of AN with detection at 4,4, and 10 ppb for margarine, honey butter, and cold-pack cheese, respectively. A peak corresponding to about 5 ppb apparent AN in all non-AN-packaged peanut butter samples examined limits detection in peanut butter to about 15 ppb. The coefficients of variation at 20 ppb for margarine, honey butter, cold-pack cheese, and peanut butter were 7.5, 8.3, 7.3, and 10.2%, respectively.
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PMID:Determination of acrylonitrile in foods by headspace gas-liquid chromatography with nitrogen-phosphorus detection. 663 Jan 23

The purpose of our study was to examine the ischemia induced enzymatic changes of decaylation-reacylation cycle of membrane phospholipids in dog brain. In this study, we developed new modified method for assay of phospholipase A1, A2 and lysophospholipase which is simpler and needs only a smaller amount of materials. For the first report, we introduced this new method and demonstrated some properties of phospholipase A1, A2 and lysophospholipase in dog brain. Crude enzyme solution for assays of phospholipase A1, A2 and lysophospholipase was gained from extraction of frozen brain with aceton, butanol and saline. The level of phosphorus in the enzyme extract was determined and only those extracts which had a level of phosphorus within a certain range were used. The substrates for assays were L-alpha-[beta-palmitoyl-1-14C] phosphatidylcholine, dipalmitoyl for phospholipase A1 and A2 and L-lysophosphatidylcholine-1-[1-14C] palmitoyl for lysophospolipase respectively. Each radioactive substrates was diluted with cold carrier lipid to give the proper specific activity. Reaction system including substrate, buffer [pH 7.0] and enzyme extract was incubated for 10 hours at 38 degrees C. But for the assay of phospholipase A1 and A2, enzyme solution was pre-incubated at 70 degrees C for 5 minutes. In our new method, reaction mixture was directly separated by TLC without extracting lipids. Enzyme activities were calculated from radio thin-layer chromatograms. Furthermore, we made a comparison between our method and the former one. The value of each enzyme activity was slightly higher in our method than in the former one. However, it was revealed that the results were reproducible in both methods.
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PMID:[Simplified microdetermination of cerebral phospholipase A1, A2 and lysophopholipase]. 663 6

The ability of the lungs to release fatty acids from circulating triglycerides or lipoproteins for its own phospholipid synthesis may be one of the factors which limit the rate of surfactant formation. Therefore, the development of the lipolytic activity of lungs obtained from late fetal and neonatal rabbits has been studied and the results correlated to the phospholipid content of lungs of similar ages. Isolated lungs were perfused with a medium which contained cold and radioactive triglyceride, and the release of fatty acids into the perfusion medium was analyzed by both colorimetric and radiochemical methods. The phospholipids of the postmitochondrial supernatant fractions of the lungs were extracted and quantified by measuring inorganic phosphorus. Finally, the influence of maternal cigarette smoke exposure on the lipolytic activity of the lungs of their litters were studied. A high lipolytic activity in the lungs of 28-day-old fetuses was detected. The activity decreased towards birth, and was lowest on the first day after birth (about 20% of that observed in 28-day-old fetuses). However, it increased again during the first week after birth. Exposure of the mothers to cigarette smoke during the last 10 days before delivery did not affect the pulmonary lipolytic activity of the offspring. Although the lung phospholipid content increased 3.6-fold from 28 days of fetal life to 1 week after birth, it remained unchanged on the days when the lung lipolytic activity was lowest. We conclude that changes in lung lipolytic activity influence lung phospholipid synthesis, and consequently influence also surfactant formation in the lungs.
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PMID:Development of the lipolytic activity in isolated perfused perinatal rabbit lungs and the influence of maternal cigarette smoke exposure on it. 663 96

Bone scintigraphy with 99 m technetium labelled phosphorus compounds was achieved in 50 west Africans migrant workers in Paris. Bone and joint tuberculosis was assumed in 20 cases. In 5 of these 20 cases, bone scan, but not X-Ray, showed abnormalities, and in 4, bone scan disclosed more localisations than X-Rays. In 7 cases, yet, bone scan was normal, with major osteolytic X-Rays lesions in 3 cases, minor in 2 cases, and isolated cold abscesses in two more cases: these means 7 false-negative results. Among the 30 other cases, 29 were considered as mechanical vertebral pathology, and 1 sacro-iliitis Brucellosis. Bone scan was normal in 28 cases the 2 others are unexplained false-positive. Although non-specific and not completely reliable, we think that bone-scanning is useful in bone-tuberculosis check-up, especially to obtain early diagnosis and detect multifocal localisations.
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PMID:[Bone scintigraphy and diagnosis of osteoarticular tuberculosis in migrant workers. Study of a series of 50 scintigraphies]. 711 56


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