Acid-soluble Phosphorus Research Articles

A simplified method for determining acetic acid soluble phosphorus in plant tissue

Abstract The proposed procedure greatly simplifies the determination of 2% HOAc soluble P in plant tissue. Extraction with Darco G 60 eliminates the need for the H2O2 oxidation of the extract. The use of vanadomolybdic acid as the chromogen results in a high degree of color stability eliminating the need for careful timing of the period between color development and transmission readings.

Acid-soluble phosphorus compounds in mammalian semen.

1. A method is described for the extraction, purification and separation of acid-soluble phosphorus compounds from mammalian semen. [8-(14)C]ATP and [8-(14)C]AMP were used as internal recovery standards to measure the breakdown and loss of these nucleotides in the procedure. 2. Bull, ram, boar and stallion semen was separated into seminal plasma and spermatozoa and the two fractions were examined separately. The overall composition of the mixture of the phosphorus compounds extracted from the two fractions was similar for the four species. 3. Glycerylphosphorylcholine and glycerylphosphorylinositol were the two phosphorus compounds identified in extracts of seminal plasma. ATP, ADP, AMP, GTP, GDP, NAD, fructose 1,6-diphosphate and glucose 6-phosphate were identified in extracts prepared from spermatozoa.

Factors Influencing Shoot Production and Mineral Nutrient Levels in Typha Latifolia

Shoot standing crops for Typha latifolia ranged from 428 to 2,252 g dry wt/m2. Standing crops were positively correlated with concentrations of dilute acid soluble phosphorus in hydrosoils and dissolved phosphorus in the waters. Except for a weak0 correlation for dissolved calcium, additional site fertility parameters were not correlated with standing crop. Tissue nutrient levels varied considerably, maximum values for most minerals being three or four times as great as the smallest values. Correlations between environmental levels of several nutrients and tissue concentrations were significant, but not very strong. Tissue concentrations of most nutrients were positively correlated with nitrogen content. Despite variations in tissue levels of nutrients, standing crop was the decisive factor determining quantities of nutrients per unit area of stand.

登熟中におけるイノシトールリン酸の変化ならびに未熟米よりイノシトールの単離

Changes of inositol phosphate and inositol in rice seeds during the ripening were studied. Total phosphorous, acid soluble phosphorus and phytin phosphorus showed a markedly rapid increase during the ripening, while inorganic phosphorus decreased. Also total inositol showed increase during ripening while free inositol decreased. Inositol phosphate were analyzed by the ion exchange chromatography (Dowex 1×2, Cl-type, 200_??_400 mesh). The main components of inositol phosphate in immature and ripe rice seeds were also inositol hexaphosphate. From 1kg of immature rice bran 533 mg of crystal was isolated by using 80% ethyl alcohol. The Rf value of crystal was 0.14 in an ascending solvent system of butanol: pyridine: H2O=6:4:3. The melting point of this crystal was at 221_??_223°C. The infrared spectrum was same with inositol standard. From these results this crystal was identified as inositol. In rice plant, it seems that inositol is synthesized at first and then inositol phosphate is produced from inositol.

The metabolism of polyphosphoinositides in hen brain and sciatic nerve.

1. The distribution of individual phospholipids was determined in hen brain and compared with that in sciatic nerve obtained in a previous investigation. Sciatic nerve is more enriched in the myelinic phospholipids ethanolamine plasmalogen, phosphatidylserine and sphingomyelin, but it contains relatively less triphosphoinositide, and much less diphosphoinositide, than the brain. 2. The course of incorporation of intraperitoneally injected (32)P into the acid-soluble phosphorus, phosphoinositides and total phospholipids of hen brain and sciatic nerve was followed. Although the maximum specific radioactivity in sciatic nerve of acid-soluble phosphorus is 4.5 times, and that of triphosphoinositide six times, that in the brain, the relative rate of triphosphoinositide phosphorus synthesis per gram of brain is three times that in sciatic nerve. 3. Administration of the demyelinating agent tri-o-cresyl phosphate to hens has no significant effect on the amounts or the rate of (32)P incorporation into the total phospholipids of the sciatic nerve. However, the rate of incorporation of (32)P into triphosphoinositide, although not its concentration, is raised from the first day after administration of the drug and remains thus 13 and 23 days later. 4. The incorporation of (32)P into polyphosphoinositides of hen brain slices in vitro was studied. The recovery of triphosphoinositide from the slices is markedly increased in the presence of EDTA, although the rate of incorporation of (32)P is unaffected. The incorporation of (32)P is dependent on the presence of Mg(2+) and Ca(2+) in the medium, and is decreased when Na(+) is replaced with K(+) or cholinium ions.

Respiration Rate and Adenosine Triphosphate Formation in Tissues4

When leaves of Nicotiana tabacum L. var. Xanthi were inoculated with TMV, local lesion formation gave enhanced ATP concentration accompanied by increased respiration. Phosphorus metabolism in healthy and infected leaves was investigated either by floating leaf tissue on 32P1-phosphate buffer or by allowing intact leaves to take up 32P1-phosphate buffer through the petioles. Both methods gave increased 32P1 incorporation into inorganic and organic acid-soluble phosphorus fractions of the infected leaf. Comparison of the specific activity of 32P1-phosphate buffer supplied with the specific activity of the γ-phosphate group of ATP demonstrated that 50 per cent of the ATP in the healthy leaf compared with over 78 per cent of the ATP in the infected leaf was metabolically active in respiration. As the ATP concentration in the infected leaf is much greater than that in the healthy leaf, this means that the amount of ATP being utilized and resynthesized as a result of respiratory metabolism in the infected leaf is more than twice that in the healthy leaf. Pulse-labelling experiments demonstrated that the rate of ATP turnover was very similar in healthy and infected leaves; therefore the increased respiration in the infected leaf results from the larger ATP pool with a turnover rate of ATP similar to that in the healthy leaf. With tobacco callus tissue cultures infected by TMV, where infection does not result in local-lesion formation, phosphorus metabolism was unaltered in the infected tissue. It is concluded that necrotic local-lesion formation results from increased availability of ATP at sites of ATP utilization, and this is aggravated by over-production of ATP in the infected leaf.

The deposition and metabolism of polyphosphoinositides in rat and guinea-pig brain during development.

1. The deposition of triphosphoinositide and diphosphoinositide in rat and guinea-pig cerebral hemispheres during growth was measured. 2. The maximum increase in concentration of both of these phospholipids occurs during the period of myelination, but in the rat some di- and tri-phosphoinositide is present before significant myelination begins. 3. In guinea-pig cerebral hemispheres the polyphosphoinositides remaining after post-mortem breakdown are selectively enriched in dissected white matter compared with grey matter. 4. The polyphosphoinositides in the cerebral hemispheres of rats were labelled with injected (32)P very rapidly; the specific radioactivities were in the order triphosphoinositide>diphosphoinositide>monophosphoinositide>total lipid phosphorus. 5. The synthesis of triphosphoinositide in rat forebrain occurs at an appreciable rate before, and at the start of, myelination, but the amount formed per gram of tissue is four to five times greater in adult rat brains, thus maintaining a constant turnover time (about 1hr.) for the whole triphosphoinositide fraction. This indicates that the rapid turnover of triphosphoinositide is independent of myelin deposition. 6. The specific radioactivity of the brain acid-soluble phosphorus pool referred to a constant dose of (32)P/g. body wt. falls rapidly with age, reaching a minimum at 13-14 days, and then rises again. The specific radioactivities of the polyphosphoinositides reflect this change. 7. Part of the polyphosphoinositides in rat and guinea-pig cerebral hemispheres is rapidly hydrolysed post mortem leaving a stable portion resistant to further breakdown. 8. The rate and extent of post-mortem hydrolysis of the polyphosphoinositides in both species decrease with age. 9. After (32)P labelling, the specific radioactivity of the triphosphoinositide remaining in the cerebral hemispheres of the rat after post-mortem breakdown is lower than the original triphosphoinositide fraction, suggesting two metabolically distinct pools.

Dose study of 32P incorporation into the nucleic acids of rat lymphoid tissues.

SummaryThere was linear incorporation of doses of 32P ranging from 0.191 to 1.91 μCi/g of body weight into DNA, RNA, and the acid soluble phosphorus fraction of the lymph nodes and thymus in rats at 4 and 48 hr after injection. The intercepts of the regression lines of these dose-incorporation curves extrapolated to 0 dose did not differ significantly from 0. These observations indicated that intraperitoneal doses of 32P up to 1.91 μCi/g of body weight did not significantly modify the rate of DNA or RNA synthesis in the lymphoid tissues of rats. There was, however, a highly significant depression of DNA synthesis in the thymus, but not in the lymph nodes 48 hr after the intraperitoneal injection of 9.5 μCi of 32P/g of body weight. It is concluded that doses of 32P up to 1.9 μCi/g of body weight may be used to study nucleic acid synthesis for time intervals up to 48 hr in the lymphoid tissues of rats.

Polyphosphates of the yeast cell vacuole.

SUMMARY: The distribution of phosphorus compounds in fractions isolated from metabolically lysed yeast protoplasts by centrifugation at 2000 g was examined. About 40% of the acid-soluble phosphorus compounds was associated with a particulate fraction which sedimented at 2000 g and which was rich in vacuoles. This phosphorus was present largely as polyphosphates of relatively high molecular weights. Evidence is presented to support the view that this polyphosphate fraction is contained in the vacuole sap. The phosphorus of the crude vacuole fraction was labelled only slowly when the protoplasts were allowed to take up 32P-orthophosphate. The phosphorus entering this fraction appeared to be derived from precursors present in the 2000 g supernatant fluid fraction.

Influence of hydrogen fluoride fumigation on acid soluble phosphorus compounds in bean seedlings

Influence of hydrogen fluoride fumigation on acid soluble phosphorus compounds in bean seedlings

A physico-chemical study of fetal calf sera used as tissue culture nutrient correlated with biological tests for toxicity

1. 1. Several physical and chemical measurements were made on a total of 96 fetal calf serum FCS samples, 84 of which were obtained under highly-standardized collection and processing procedures at cooperating slaughter houses in Omaha, Nebraska, and in St. Paul, Minnesota. Other FCS samples, and four non-FCS samples, were measured either as controls or for comparison. 2. 2. Biological tests were carried out using individual FCS samples as a part of the culture medium with MDBK, HeLa, and primary dog kidney cells. 3. 3. Toxicity was demonstrated in two control FCS samples known to be toxic, and one of the commercial FCS samples. Limited biological efficacy was observed in three of the FCS samples obtained from cooperating slaughter houses. All other FCS samples allowed cellular attachment and growth to take place in biological tests. 4. 4. The chemical parameters that correlated with toxicity in FCS included a high total protein content, an abnormality in the protein electrophoretic pattern, low acid soluble phosphorus content, and a high phospholipid and total cholesterol content. 5. 5. Total protein showed an increase in FCS with increasing age of the fetus regardless of breed or geographical location of the sampling. 6. 6. Other physical and chemical parameters that were measured included specific gravity, refractive index, pH, A G ratios Na +, Cl −, Ca 2+, Ca P ratios, glucose, hemoglobin, urea nitrogen, and total lipid content. These measurements were within expected normal values for FCS and did not correlate with biological tests.

An assay for organic phosphorus fractions in microgram quantities of tissue

A fluorometric technique for the determination of acid-soluble phosphorus, lipid phosphorus, and nucleic acid phosphorus in 0.5 to 1.0 μg quantities of tissue is described. The assay is based on the stoichiometric generation of NADPH from P i by the use of purified crystalline enzymes, a feature which confers the advantage of extension to even greater sensitivity by application of the “NADPH cycling” system. Assays of mouse brain and mouse brain tumor were performed at both the microgram and milligram level and compared. The fluorometric assay offers about fivefold increase in sensitivity with a sacrifice of analytical accuracy of approximately 6% (4% vs. 10%) when compared to an established spectrophotometric procedure.

Effect of Ion Exchange Resins on Composition of Milk and Its Fractions

Ultracentrifugation and dialysis studies were made to determine the distribution of calcium, phosphorus, and nitrogen in milk treated with ion exchange resins to remove radionuclides. In System I, milk was passed through Dowex 2-X8 (Cl :P :Cit) resin, acidified to pH 5.35 with citric acid, passed through IR-120 (Ca:Mg:K:Na) resin, and neutralized to pH 6.8 with KOH. In System II, the same cycle was followed, except that HCl was used to acidify milk and the milk was neutralized with IRA-68 (P:Cit:OH) weak base anionic resin.A comparison of the test results for treated samples and those for the control sample showed increased concentrations in the casein nitrogen and in acid soluble phosphorus in serum, and a decreased amount of sedimented casein. The amount of calcium and acid soluble phosphorus associated with the sedimented caseinates also decreased. The extent of these alterations depended upon the type of acid used to acidify the milk. The effect of these changes in the ionic balance on the coagulation of milk by rennin and ethanol is briefly discussed.

Tissue distribution and transport of electrolytes Mg 28 and Ca 47 in hypermagnesemia

The effect of acute hypermagnesemia (9–14 mEq./L.) on the distribution of electrolytes Mg 28 and Ca 47 in various tissues was studied in renal-ligated dogs. The sodium content of 5 of 8 tissues was decreased by 10–30 per cent, the potassium content of 3 tissues was increased 5–30 per cent and the acid-soluble phosphorus concentration of 2 tissues was decreased by 15 per cent. Total and/or exchangeable cellular concentrations of calcium were decreased by 25–35 per cent in skeletal muscle, myocardium, brain and pancreas. Total and/or exchangeable cellular concentrations of magnesium were increased by 8–50 per cent in skeletal muscle, nerve, pancreas and aorta and were unchanged in liver, myocardium and brain. These data suggest that magnesium has inhibitory effects on the cellular transport of electrolytes which are similar to those of calcium. The cellular influx of calcium is inhibited by magnesium. The stability of the cellular concentrations of magnesium in relation to the degree of hypermagnesemia is compatible with in vitro studies in rat liver indicating that the fractional rate of cellular influx of magnesium is decreased and the fractional rate of cellular efflux of magnesium is increased during extreme hypermagnesemia.

1,2-Propanediol-2-Phosphate in Ascaris lumbricoides

No phosphagen is present in the body-wall muscle of Ascaris lumbricoides. A stable phosphorus compound, which on the basis of its chemical reactions seems to be 1,2-proepanediol-2-phosphate, constitutes more than half of the acid-soluble phosphorus compounds.

1,2-Propanediol-2-Phosphate in Ascaris lumbricoides

No phosphagen is present in the body-wall muscle of Ascaris lumbricoides . A stable phosphorus compound, which on the basis of its chemical reactions seems to be 1,2-propanediol-2-phosphate, constitutes more than half of the acid-soluble phosphorus compounds.

Effect of vitamin D on tissue distribution and transport of electrolytes, 47-Ca and 28-Mg.

The administration of 100,000 to 200,000 U of calciferol to adult dogs daily for 5 weeks caused marked hypercalcemia and significant decreases in plasma concentrations of sodium, potassium, magnesium and chloride. Cellular concentrations of calcium were increased by 30 to 300 % in muscle, myocardium, pancreas and aorta and were unchanged in liver, brain and nerve. Bone uptake of 47Ca was increased by 45 %. The exchangeable body and tissue pools of calcium were increased by 100% but the fractional rate of exchange of tissue calcium was unaltered. The magnesium concentrations of liver, pancreas and bone were increased by 16 to 20 % by calciferol and the fractional rate of exchange of tissue magnesium with 28Mg was increased by 23 %. Calciferol administration had a minor effect on tissue concentrations of acidsoluble phosphorus. The sodium concentrations of 5 tissues and the chloride concentrations of 3 tissues were significantly decreased, whereas the potassium concentrations of 2 tissues were increased sig...

The effect of temperature, anoxia, or injection of various substances on haemolymph composition and supercooling in larvae of Anagasta kuehniella (Zell.)

The supercooling and freezing points of last-instar larvae of the Mediterranean flour moth ( Anagasta kuehniella (Zell.)) were highest in larvae stored at 20°C, and were gradually lowered after storage at 6°, 0°, or −6°C. Depressions of supercooling points were, however, much larger than those of freezing points. Concurrent with this increase in amount of supercooling, an increase in concentrations of several haemolymph solutes was found. A rise in total content of ninhydrin-positive substances was mainly due to an increase in alanine content. The content of glycerol and glucose also increased at low temperatures. Concentrations of trehalose increased during storage at 0°C, but decreased at −6°. There were also some increases in content of acid-soluble phosphorus compounds, in particular of inorganic phosphorus. Anoxia for 12 or 24 hr also resulted in an increased amount of supercooling, concurrent with increases in the haemolymph of ninhydrin-positive substances, glycerol, glucose, lactate, and inorganic phosphorus. The concentration of trehalose was lowered during anoxia. When present the concentrations of glycerol were relatively low, and it was absent or almost absent in larvae exposed to 6°C or to 12 hr of anoxia. It is therefore concluded that the observed increases in amount of supercooling are probably an effect of increases in concentrations of several haemolymph solutes. Oral injection of glucose and trehalose further demonstrated that substances other than glycerol may affect supercooling. Oral injection of glycerol clearly showed a relation between this substance and supercooling. A rise in supercooling points following injection of l-alanine could not be explained.

Composition of bull epididymal and testicular fluid.

<section class="abstract"><h2 class="abstractTitle text-title my-1" id="d549896e2">Summary.</h2> Analyses were made of fluid collected from the rete testis and the caput, corpus and cauda epididymidis of the bull. Both the major electrolytes and some organic constituents in the fluids were examined. There was a decrease in sodium and chloride as the fluid passed from the testis through the epididymis and at the same time there was a general increase in organic constituents. In testicular fluid, sodium and chloride made up the bulk of the ions present. In the epididymis, however, potassium replaced approximately half of the sodium and at the same time there was a general decrease in total electrolytes. Of the organic constituents, orcinol-reactive carbohydrate, protein and acidsoluble phosphorus, especially glycerylphosphorylcholine, were present in much higher concentrations in the epididymal than in the testicular fluid. Although little reducing sugar was found, lactic acid was present in all the fluids examined. </section>

Glycolytic metabolites and their distribution at death in the white and red muscle of cod following various degrees of antemortem muscular activity.

Muscular activity just before death critically affected the levels of glycolytic metabolites in the skeletal muscle of feeding, aquarium-held cod. Relaxed cod could be obtained by slow anesthetization without visible excitation; these contained very little lactate in the white muscle (7–28 mg/100 g), and exhibited high, uniformly distributed levels of high-energy phosphate compounds (about 17 μmoles acid-labile P/g plus equal or higher amounts of creatine phosphate). A trend to lower glycogen levels in the anterior portion of the fillet was indicated. Unexercised fish caught and killed with negligible struggling appeared to have undergone some antemortem activity. Compared with relaxed cod, they showed higher lactate levels (about 100 mg/100 g), much lower levels of creatine phosphate and other high-energy phosphate compounds, together with some ammonia. Again, glycogen was lower in the anterior than in the posterior portion. Exhaustion uniformly depleted energy reserves to low levels in the white muscle; in the red muscle, however, high glycogen levels were maintained, although the content of other glycolytic metabolites indicated partial activation of the glycolytic processes during the antemortem struggle. In all fish examined, levels of inorganic and total acid-soluble phosphorus were lower in the red than in the white muscle.