Phosphorus-31 Nuclear Magnetic Resonance Spectra Research Articles

The Limitation of Unproductive Binding of Cellulases to Lignin by Ozone Pretreatment

The limitation of enzymatic saccharification of lignocellulose is attributed to the nonproductive adsorption between lignin and cellulase. This study aims to investigate the effects of ozone pretreatment on the physical structure and chemical properties of milled wood lignin (MWL). The objective is to reduce the non-productive adsorption of cellulase on lignin. The structure–activity relationship between the physical structure of MWL and the occurrence of nonproductive adsorption was analysed using two-dimensional heteronuclear single quantum coherence–nuclear magnetic resonance (HSQC-NMR) and phosphorus-31 nuclear magnetic resonance spectrum (31P-NMR), etc. The results indicate that ozone pretreatment resulted in a decrease in the phenolic hydroxyl content and S/G ratio, an increase in the carboxyl content, and a negative zeta potential of MWL. The maximum adsorption capacity decreased from 25.77 mg/g to 10.09 mg/g, the Langmuir constant decreased from 13.86 mL/mg to 10.11 mL/mg, and the binding strength decreased from 357.14 mL/g to 102.04 mL/g, as determined by Langmuir isothermal adsorption. This suggests that ozone pretreatment resulted in a reduction in the hydrophobicity of lignin and a weakening of the electrostatic attraction between lignin and cellulase, thereby effectively reducing the non-productive adsorption of cellulase on lignin. This study provides an environmentally friendly pretreatment technique and comprehensively analyses the structural changes of ozone-treated MWL. These findings contribute to a deeper understanding of the interaction between lignin and cellulase.

ATP Analogues for Structural Investigations: Case Studies of a DnaB Helicase and an ABC Transporter.

Nucleoside triphosphates (NTPs) are used as chemical energy source in a variety of cell systems. Structural snapshots along the NTP hydrolysis reaction coordinate are typically obtained by adding stable, nonhydrolyzable adenosine triphosphate (ATP) -analogues to the proteins, with the goal to arrest a state that mimics as closely as possible a physiologically relevant state, e.g., the pre-hydrolytic, transition and post-hydrolytic states. We here present the lessons learned on two distinct ATPases on the best use and unexpected pitfalls observed for different analogues. The proteins investigated are the bacterial DnaB helicase from Helicobacter pylori and the multidrug ATP binding cassette (ABC) transporter BmrA from Bacillus subtilis, both belonging to the same division of P-loop fold NTPases. We review the magnetic-resonance strategies which can be of use to probe the binding of the ATP-mimics, and present carbon-13, phosphorus-31, and vanadium-51 solid-state nuclear magnetic resonance (NMR) spectra of the proteins or the bound molecules to unravel conformational and dynamic changes upon binding of the ATP-mimics. Electron paramagnetic resonance (EPR), and in particular W-band electron-electron double resonance (ELDOR)-detected NMR, is of complementary use to assess binding of vanadate. We discuss which analogues best mimic the different hydrolysis states for the DnaB helicase and the ABC transporter BmrA. These might be relevant also to structural and functional studies of other NTPases.

Homogeneous and heterogeneous photocatalytic water oxidation by polyoxometalates containing the most earth-abundant transition metal, iron

Reported here, inorganic molecular tetrairon(III)-substituted polyoxotungstate [FeIII4(H2O)2(P2W15O56)2]12− is firstly used to catalyze visible-light-driven oxygen evolution, which displays excellent catalytic activity (O2 yield of 48%, turnover number (TONcatalyst) of 900 and quantum (ΦQY) yield of 24%) in a homogeneous system. The O2 yield and TON of Fe4POM catalyst rank third and fourth among thirty documented homogeneous photocatalytic water oxidation catalysts based on POMs, respectively. In order to recover and reuse the homogeneous catalysts, it is realistic and necessary to heterogenize the homogeneous molecular catalyst. The electrostatic interaction is rationally designed as solid connection between ((3-aminopropyl)triethoxysilane) apts-modified mesoporous SBA-15 and polyoxoanion (Fe4POM). The catalytically active species of immobilized polyoxometalate maintains intact before and after the photocatalytic water oxidation, which is adequately confirmed by various spectroscopic techniques such as solid phosphorus-31 nuclear magnetic resonance spectra (31P MAS NMR), X-ray photoelectron spectra (XPS) and transmission electron microscopy (TEM) images.

Identification of Phytate in Phosphorus-31 Nuclear Magnetic Resonance Spectra: The Need for Spiking

Phosphorus-31 (31P) nuclear magnetic resonance (NMR) spectroscopy of sodium hydroxide–ethylenediaminetetra-acetic acid (NaOH-EDTA) extracts has recently become a widely used technique for the characterization of soil P. This technique has seemingly enabled easy identification and quantification of phytate (myo-inositol hexakisphosphate), a compound long believed to constitute a major proportion of organic P. Phytate is usually identified by its characteristic pattern of four resonances in the ratio 1:2:2:1. We report that the 31P NMR spectra of the NaOH-EDTA extracts of four Australian pasture soils contain a set of resonances that bear a striking resemblance to the phytate resonances but that are shown not to be phytate though careful addition (spiking) of pure phytate. The spiking experiments identify a much smaller set of resonances as being phytate. Quantification of these resonances shows that phytate comprises <5% of organic P and <3% of total P in these soils. We also show that the 31P chemical shift of phytate resonances is very sensitive to pH and ionic strength. These results highlight the potential for misassignment of resonances in 31P NMR spectra of NaOH-EDTA extracts of soil and the possibility that phytate concentrations may be overestimated using this technique and show the value of spiking as a definitive form of identification of 31P NMR resonances.

Proton and phosphorus-31 nuclear magnetic resonance spectroscopy of human bile in hepatopancreaticobiliary cancer

Hepatopancreaticobiliary cancers can be difficult to diagnose. Nuclear magnetic resonance (NMR) spectroscopy provides non-invasive information on phospholipid metabolism, and previous studies of liver tissue have highlighted changes in phospholipids in malignancy. We hypothesised that in-vitro NMR spectroscopy of human bile may provide independent diagnostic indices in cancer management through an assessment of the phospholipid content. Bile samples from 24 patients were collected at endoscopic retrograde cholangiopancreatography and from one subject at cholecystectomy. Thirteen patients had cancer: pancreatic carcinoma (eight), cholangiocarcinoma (three) and metastatic liver disease (two). The remaining 12 patients had non-malignant pathology. In-vitro proton (H) and phosphorus-31 (P) NMR spectra were obtained from all samples using an 11.7 Tesla NMR spectroscopy system. Complementary information was obtained from the H and P NMR spectra. Signals were assigned to phosphatidylcholine in both H and P NMR spectra. Phosphatidylcholine levels were significantly reduced in the bile from cancer patients when compared with bile from non-cancer patients (P=0.007). These preliminary studies suggest that H and P NMR spectroscopy of bile may be used to detect differences in phospholipid content between cancer and non-cancer patients. This may have implications for the development of novel diagnostic strategies in hepatopancreaticobiliary cancers. Further larger-scale studies are warranted.

Phosphorus-31 nuclear magnetic resonance study of the effect of pentachlorophenol (PCP) on the physiologies of PCP-degrading microorganisms.

Free and agarose-encapsulated pentachlorophenol (PCP)-degrading Sphingomonas sp. isolates UG25 and UG30 were compared to Sphingomonas chlorophenolica ATCC 39723 with respect to the ability to degrade PCP. Pretreatment of the UG25 and UG30 strains with 50 microg of PCP per ml enabled the cells to subsequently degrade higher levels of this environmental pollutant. Similar treatment of ATCC 39723 cells had no effect on the level of PCP degraded by this strain. Phosphorus-31 nuclear magnetic resonance spectra of agarose-immobilized strains UG25 and UG30 grown in the absence of PCP showed that there was marked deenergization of the cells upon exposure to a nonlethal concentration of PCP (120 microg/ml). For example, no transmembrane pH gradient was observed, and the ATP levels were lower than the levels obtained in the absence of PCP. The transmembrane pH gradient and ATP levels were restored once the immobilized cells had almost completely degraded the PCP in the perfusion medium. PCP-pretreated cells, on the other hand, maintained their transmembrane pH gradient and ATP levels even in the presence of high levels of PCP. The ability of PCP-pretreated strain UG25 and UG30 cells to remain energized in the presence of PCP was shown to correlate with an altered membrane phospholipid profile; these cells had a higher concentration of cardiolipin than cells cultured in the absence of PCP. Strain ATCC 39723, which did not degrade higher levels of PCP after PCP pretreatment, did not show this response.

Differential uptake of FDG and DG during post-ischaemic reperfusion in the isolated, perfused rat heart.

Fluorine-18 2-fluoro-2-deoxyglucose (FDG) and 2-deoxyglucose (DG) are widely used as tracers of glucose uptake in the myocardium. Although there is agreement that the two analogues behave similarly to glucose under control conditions, there is growing evidence that some interventions (e.g. insulin stimulation or ischaemia/reperfusion) cause differential changes in their behaviour. The addition of a two-surface coil nuclear magnetic resonance (NMR) probe and a dual-perfusion cannula to our recently developed PET and NMR dual-acquisition (PANDA) system allows us to collect PET (FDG) images and phosphorus-31 NMR (2-deoxyglucose-6-phosphate) spectra simultaneously from each independently perfused coronary bed of the heart. We have used this technique to study the effect of regional ischaemia/reperfusion on FDG and DG uptake in the isolated, perfused rat heart. During control perfusion, FDG uptake was almost identical in both coronary beds. When one coronary bed was made ischaemic, FDG uptake ceased on that side but continued on the control side. Reperfusion failed to restore FDG uptake. In contrast, NMR spectra showed that, during reperfusion, the uptake and phosphorylation of DG did not differ between the two coronary beds. The results thus demonstrate that regional myocardial ischaemia/reperfusion has different effects on the uptake of FDG and DG in the isolated, perfused rat heart.

Phosphorus-31 magnetic resonance spectroscopy studies of pig spinal cord injury. Myelin changes, intracellular pH, and bioenergetics.

Phosphorus-31 (31P) nuclear magnetic resonance (NMR) spectroscopy was used to monitor changes in phosphocreatine (PCr), adenosine triphosphate (ATP), inorganic phosphate (Pi), intracellular pH (pHi), and free magnesium in the in vivo pig spinal cord after injury. Phosphorus-31 NMR spectra were acquired from healthy (n = 4) and injured pig spinal cords (n = 8) under in vivo conditions using a 4.7-tesla spectrometer. Spinal cords were injured by dropping a 20-g weight from 20 cm onto the surgically exposed cord surface. In vivo spectra of injured cords revealed a reduction in ATP, PCr, pHi, and an increase in Pi. In addition, a broad resonance that is likely to arise from myelin phospholipids was reduced significantly after injury. Phosphorus-31 NMR can be used to follow in vivo changes in high energy phosphates after injury and may have the potential to follow changes in myelin structure. This technique may prove important in the study of myelin breakdown after secondary, nonreversible spinal cord injury. Changes in high energy phosphates and pHi did not seem to parallel these putative changes in myelin structure.

Effects of thyroid hormone treatment on 31P-NMR spectroscopy of muscle and on nerve conduction studies in a patient with long-standing severe hypothyroidism

Hypothyroidism exerts a significant effect on several body systems, including the functioning of muscle and nerve. Described here is the case of a severely hypothyroid patient with weakness and numbness of the legs who showed marked clinical improvement with thyroid hormone treatment. Concurrently obtained phosphorus 31-nuclear magnetic resonance (NRM) spectra and nerve conduction studies showed changes in muscle phosphorus metabolism with increases in conduction velocities. Phosphorus 31-NMR spectroscopy is useful for evaluating metabolism in muscle in hypothyroid patients.

Phosphorus-31 nuclear magnetic resonance spectroscopy of the spinal cord in the pig, rat, and rabbit.

To ensure that contamination-free phosphorus-31 nuclear magnetic resonance (31P-NMR) spectra of the spinal cord could be obtained, a porcine model was adopted that provided a large cord sample and a greater area free from adjacent muscle tissue. Phosphorus-31 NMR spectra were acquired from the porcine spinal cord under in vivo conditions using a 4.7-T spectrometer. Spectra also were collected from perchloric acid and lipid extracts, and excised freeze trapped samples of the rat, rabbit, and pig spinal cord. The in vivo spectrum showed resonances corresponding to adenosine triphosphate, phosphocreatine, inorganic phosphate, phosphomonoesterase, and phosphodiesterase as confirmed by extracts. In addition, a broad resonance was observed that was assigned to myelin phospholipids. Phosphorus-31 NMR spectra of the spinal cord revealed resonances common to brain tissue. Importantly, the existence of a previously undetected resonance, which is likely to correspond to myelin phospholipids, also is reported. This resonance may prove important in future studies monitoring changes in myelin in response to trauma and ischemia.

Water-soluble reserve polysaccharides fromAchlyaare (1→3)-β-glucans

The cytoplasmic water-soluble p-glucans of Achlya bisexualis are (1?>3)-linked polysaccharides of two types: a small neutral and a large phosphorylat- ed. The carbon-13 nuclear magnetic resonance (NMR) spectrum of neutral glucan is consistent with a (l-^3)-(3-glucan with some (1?>6)-(3-linkages. Meth- ylation analysis suggests a similar structure, but also shows about 6% each of other linkages and other sugars. Acid and enzyme hydrolysis data also support a moderately branched (1??3)-p-glucan probably with single (1?>6)-p-glucopyranosyl segments. The data, however, do not exclude the possibility that the branches are extended (1?>3)-linked residues. The molecular mass of neutral glucan is approximately 9000 with a dp of 50, when compared with algal 1am- inaran by gel filtration chromatography. The carbon- 13 NMR spectrum of phosphoglucan is qualitatively similar to that of neutral glucan. Aqueous hydrofluor- ic acid effectively cleaves phosphate from the phos? phoglucan and produces a polymer of the size of neutral glucan. This material is both qualitatively and quantitatively identical to neutral glucan in its car? bon-13 NMR spectrum. The phosphorus-31 NMR spectrum of the phosphoglucan reveals both mono- and diphosphoester linkages. We propose that the phosphodiester covalent bonds convert the smaller polymer into the very large phosphoglucan.

Effects of the Mycelial Extract of Cultured Cordyceps Sinensis on In Vivo Hepatic Energy Metabolism in the Mouse

Mice were given the extract of cultured Cordyceps sinensis (Cs) (200 mg/kg daily, p.o.) for 3 weeks. In vivo phosphorus-31 nuclear magnetic resonance (NMR) spectra of the liver were acquired at weekly intervals using a surface coil. From 1 to 3 weeks, a consistent increase in the ATP/inorganic phosphate ratio, which represents the high energy state, was observed in the Cs extract-treated mice. The intracellular pH of the Cs extract-treated mice was not significantly different from that of the control mice. No steatosis, necrosis, inflammation or fibrosis were observed in the liver specimens from Cs extract-treated mice.

A 31P-Nuclear magnetic resonance study of intermittent warm blood cardioplegia

This study was designed to assess the effects of intermittent warm blood cardioplegia on myocardial energy metabolites, intracellular pH, and contractile function. The isolated blood-perfused pig hearts were divided into three groups. After 30 minutes of control perfusion, the hearts in group 1 ( n = 10) received 90 minutes of continuous warm (37° C) blood cardioplegia; the hearts in group 2 ( n = 9) received six 5-minute periods of warm blood cardioplegia, interrupted by six 10-minute episodes of ischemia (37° C). The hearts were then reperfused for 30 minutes. The hearts in group 3 underwent 150 minutes of control perfusion without cardioplegia or ischemic episodes. Phosphorus 31-nuclear magnetic resonance spectra showed that a 10-minute interruption of warm blood cardioplegia decreased phosphocreatine levels and intracellular pH by approximately 47% ( p < 0.01) and 0.12 unit ( p < 0.05), respectively, and increased inorganic phosphate levels by approximately 87%, whereas resumption of cardioplegia for 5 minutes resulted in almost 100% recovery of phosphocreatine and inorganic phosphate levels and intracellular pH. More important, subsequent interruptions did not result in any cumulative changes in phosphocreatine level, inorganic phosphate level, or intracellular pH beyond those changes observed after the initial cardioplegic interruption. Moreover, during reperfusion there were no significant differences in adenosine triphosphate and phosphocreatine levels among the three groups of hearts. Furthermore, hearts from groups 1 and 2 showed comparable recovery of contractile function. These results indicate that six 10-minute interruptions and six 5-minute restorations of warm blood cardioplegia caused only mild and reversible changes in myocardial energy metabolites and intracellular pH and these changes were not cumulative. This study suggests that antegrade intermittent warm blood cardioplegia may provide as much myocardial protection as does antegrade continuous warm blood cardioplegia in the normal heart . (J T HORAC C ARDIOVASC S URG 1995;109:1155-63)

Phosphate metabolites of Tenebrio molitor (Coleoptera: tenebrionidae) infected with metacestodes of Hymenolepis diminuta.

In vivo phosphorus-31 nuclear magnetic resonance (31P NMR) spectra and those of extracts of Tenebrio molitor L. infected with metacestodes of Hymenolepis diminuta R. showed modifications in the amounts of phosphorous-containing metabolites when compared with those of uninfected beetles. Infected females were more affected than infected males, having significantly more glucose-6-phosphate (Glu-6-P), glycerol-3-phosphate (Gly-3-P), and phosphorylethanolamine (PE), but less inorganic orthophosphate (Pi), glycerophosphorylethanolamine (GPE), gylcerolphosphorylcholine (GPC), phosphoarginine (PAr), and adenosine diphosphate (ADP). Infected males had more Glu-6-P and PE, but less Pi, GPE, and GPC. These changes directly reflected the phosphorylation potential of infected hosts, which increased approximately 100% and 50% in infected females and males, respectively.

31P nuclear magnetic resonance spectra and dissociation constants of lac repressor headpiece.duplex operator complexes: the importance of phosphate backbone flexibility in protein.DNA recognition.

An alkaline phosphatase assay was used to determine the dissociation constants (KD) of the lac repressor N-terminal 56 amino acid fragment of the wild type and of a Y7I mutant complexed to 22 base pair (bp) wild-type and mutant symmetrical operator sequences. KD's in 0.35 M monovalent salt ranged from 5.4 X 10(-8) M for the wild-type repressor.wild-type operator complex to approximately > 1 X 10(-6) M for the wild-type repressor.nonspecific DNA complex. Mutant operators O2 (G5 --> A5 and G16 --> T16) and O4 (G5 --> C5 and C16 --> G16) bind nearly as tightly as the wild-type headpiece, while mutant O3 (A8 --> T8 and T13 --> A13) binds over 5-fold poorer. Operators O1, O2, and O4 bind ca. 10-fold poorer to the Y7I mutant headpiece. Operator O3 binds 2-fold poorer to the mutant headpiece. The temperature and salt dependence on the dissociation constants of wild-type headpiece binding to 22-bp operator support the conclusion that the headpiece contains the major DNA recognition portion of the protein and that electrostatics plays as important a role in the binding of operator to headpiece as it does in the whole repressor. The 31P NMR spectra of shortened 14-bp wild-type and mutant symmetrical operators bound to the N-terminal 56-residue headpiece of the Y7I mutant repressor were compared to the spectra of the same operator bound to the wild-type repressor headpiece. These results are consistent with a recent proposal [Karslake, C., Botuyan, M. V., & Gorenstein, D. G. (1992) Biochemistry 31, 1849-1858] that specific, tight-binding operator.protein complexes retain the inherent phosphate ester conformational flexibility of the operator itself, whereas the phosphate esters are conformationally restricted in the weak-binding operator-protein complexes. This retention of backbone torsional freedom in tight complexes is entropically favorable and provides a mechanism for protein discrimination of different operator binding sites.

Role of sodium/calcium exchange in the mechanism of myocardial stunning: Protective effect of reperfusion with high sodium solution

Objectives. This study was conducted to elucidate the role of sodium/calcium (Na+/Ca2+) exchange in the mechanism of myocardial stunning.Background. Cellular Ca2+overload mediated by Na+/Ca2+exchange during reperfusion has been proposed as a mechanism for myocardial stunning. Because no specific pharmacologic inhibitors of the exchanger are available, we increased extracellular sodium concentration ([Na]0) during the early phase of reperfusion to decrease the driving force for Ca2+influx through the pathway.Methods. Isovolumetric left ventricular pressure and phosphorus-31 nuclear magnetic resonance spectra were measured in isolated perfused ferret hearts. Hearts were reperfused with different solutions after 15 min of total global ischemia at 37 °C.Results. Hearts reperfused with standard solution ([Na]0= 140 mmol/liter; the stunned hearts, n = 8) showed only 69 ± 3% (mean ± SEM) recovery of developed pressure relative to preischemic control developed pressure, In contrast, hearts reperfused with a high [Na]0solution ([Na]0= 268 mmol/liter) during the initial 5 min, followed by a gradual decrease of [Na]0to the standard level over 25 min (the high [Na]0group, n = 8) showed significantly better recovery of developed pressure (85 ± 2%, p < 0.05 vs. the stunned hearts). in contrast, reperfusion with solutions in which the additional Na was substituted either by 256 mmol/liter sucrose or 128 mmol/liter chollne chloride did not improve functional recovery, indicating that the beneficial effects of high [Na]0reperfusion are not due to either high ionic strength or high osmolarity. Phosphorus-31 nuclear magnetic resonance spectra showed no correlation between functional recovery and intramyocardial contents of phosphorus compounds or pH.Conclusions. High [Na]0reperfusion protects against stunning, supporting the concept that Na+/Ca2+exchange plays an important role in the mechanism of stunned myocardium.

Phosphorus-31 nuclear magnetic resonance study on the effects of endurance training in rat skeletal muscle.

To evaluate changes in muscle energetics following endurance training, we measured phosphorus-31 nuclear magnetic resonance (31P NMR) spectra on rat muscle in vivo before and after training in the same animals. The endurance training lasted for 3 months. The 31P NMR spectra were obtained serially at rest, during exercise by electrical stimulation, and during recovery. Intramuscular phosphocreatine (PCr), inorganic phosphate (P(i)), adenosine 5'-triphosphate (ATP) and pH were determined from the NMR spectra. The ratio of PCr:(PCR + P(i) at rest showed no difference between the trained and control groups even after 3 months of training. During exercise, however, this ratio was significantly higher in the trained group than in the control group. The ratio also recovered more rapidly after exercise in the trained group. The intramuscular pH decreased slightly by approximately 0.1 pH unit during exercise but did not show a significant difference between the groups. These results indicated that endurance training of 3 months duration improved the ATP supply system in the muscle. They also demonstrated that 31P NMR is a potent method for evaluating the effects of training in the same individuals.

Response of the headgroup of phosphatidylglycerol to membrane surface charge as studied by deuterium and phosphorus-31 nuclear magnetic resonance

The response to membrane surface charge of the glycerol headgroup of dimyristoyl-phosphatidylglycerol (DMPG) was investigated via deuterium and phosphorus-31 nuclear magnetic resonance spectroscopy. The membrane surface charge was manipulated by adding various amounts of neutral dimyristoylphosphatidylcholine (DMPC) and/or positively charged didodecyldimethylammonium bromide (DDAB) to the negatively charged DMPG, selectively deuterated at the alpha and beta segments of its glycerol headgroup. The deuterium and phosphorus-31 nuclear magnetic resonance spectra were all characteristic of random dispersions of liquid-crystalline lipids in a bilayer configuration. Differential scanning calorimetry showed that all mixtures investigated exhibited gel to liquid-crystalline phase transitions below 35 degrees C. Measurements of the deuterium quadrupole splitting and of the phosphorus-31 chemical shift anisotropy lead to the following observations. (1) Dilution of the negative surface charge density by the addition of DMPC had little effect on the quadrupole splitting from either alpha- or beta-deuterated DMPG. (2) Direct cancellation of the negative surface charge density by addition of DDAB led to a progressive decrease in the quadrupole splitting measured from alpha-deuterated DMPG, while the quadrupole splitting measured from beta-deuterated DMPG increased. For alpha-deuterated DMPG addition of 0.3 mole fraction of DDAB resulted in the appearance of two distinct quadrupole splittings. No such effect was observed for beta-deuterated DMPG.

Effect of cyclosporine on hepatic energy status and on fructose metabolism after portacaval shunt in dog as monitored by phosphorus-31 nuclear magnetic resonance spectroscopyin vivo

The effect of cyclosporin A on the hepatic energy status and intracellular pH of the liver and its response to a fructose challenge has been investigated using in vivo phosphorus-31 nuclear magnetic resonance spectroscopy in dogs. Three experimental groups were studied: (a) control dogs (n = 5), (b) dogs 4 days after the creation of an end-to-side portacaval shunt (n = 5), and (c) dogs 4 days after portacaval shunt and continuous infusion of cyclosporin A (4 mg/kg/day) by way of the left portal vein (portacaval shunt plus cyclosporin A, n = 5). The phosphorus-31 nuclear magnetic resonance spectra were obtained at 81 MHz using a Bruker BIOSPEC II 4.7-tesla nuclear magnetic resonance system equipped with a 40-cm horizontal bore superconducting solenoid. The phosphomonoesters (p less than 0.01), inorganic phosphate and ATP levels (p less than 0.05) were decreased significantly in portacaval shunt-treated and in portacaval shunt-plus-cyclosporin A-treated dogs compared with unshunted control dogs. After a fructose challenge (750 mg/kg body wt, intravenously), fructose-1-phosphate metabolism was reduced in portacaval shunt-treated dogs compared with either the normal or portacaval shunt-plus-cyclosporin A-treated dogs (p less than 0.05). Both portacaval shunt- and portacaval shunt-plus-cyclosporin A-treated dogs demonstrated a reduced decline in ATP levels after fructose infusion when compared with the controls (p less than 0.05). Immediately after the fructose challenge, the intracellular pH decreased from 7.30 +/- 0.03 to 7.00 +/- 0.05 in all animals (p less than 0.01) and then gradually returned to normal over 60 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Phosphorus-31 and selenium-77 nuclear magnetic resonance spectra of phosphorus-poor phosphorous selenide compounds

The compounds P3Se4X (X = I, Br, Cl, SC2H5, SC5H11 or SC6H5) have been made as phosphorus-poor analogues of the corresponding compounds α- or β-P4E3X2(E = S or Se), in which a PX unit has been replaced by Se. Analysis of 31P and (in part)77Se NMR spectra of the P3Se4X compounds has shown remarkable correspondence of the coupling constants and chemical shifts between the P3Se4X and α-P4S3X2 series of compounds. Phosphorus-31 and 77Se NMR spectra of α-P4Se3I2 are reported for comparison. Further replacement of a PX unit by Se gives the ultimate phosphorus-poor cluster compound P2Se5, for which a synthesis from the elements and full NMR data are reported. This sought after higher selenide of phosphorus is thus a phosphorus-poor molecule with concatenated selenium, rather than a selenium-rich structure with exocyclic selenium. The interconversion of optical isomers of the P3Se4X halides (X = I, Br or Cl), by skeletal rearrangement on the NMR time-scale, has been studied at several temperatures, yielding kinetic data.