Ammonium and phosphate regenerationby the zooplankton of an Amazon floodplain lake

SUMMARY. 1. Regeneration of ammonium and phosphate by macro-zooplankton (Cladocera. adult copepods. and copepodites) was measured in Lake Calado. an Amazon floodplain lake, Macrozooplanktonabundances ranged between 1×10⁴ and 3×10⁵ individuals m⁻².
2. Phosphate regeneration ranged from 0.2 to 1.3 μ mol PO₄ m⁻² b⁻¹at station 1. located 2 km from the Solimoes River, and from 1.6 to8.3 μ mol PO₄ m⁻² h ⁻¹ at station 3, located 7 km from the SolimoesRiver. Ammonium regeneration at stations 1 and 3 ranged from 1.7 to11.9 and from 13.4 to 77.2 μ mol NH₄ m⁻² h⁻¹. respectively.
3. Zooplankton regenerated ammonium and phosphate at similarrates during rising and falling waier. Regeneration by macrozooplankton was low compared to other tropical lakes and compared to microbesand microzooplankton in Lake Calado.     

Microbial activity associated with seston in headwater streams: effects of nitrogen, phosphorus and temperature

SUMMARY 1. The influences of temperature and dissolved nitrates and phosphates on microbial activity associated with suspended fine particulate organic matter (seston) were evaluated in four headwater streams in the southern Appalachian Mountains.
2. Temperature manipulations of ± 5°C always induced significant changes in [¹⁴C] glucose mineralization (ANOVA; P <0.05) and [³H]thymidine incorporation (ANOVA; P <0.05).
3. Nutrient amendments of 1.0 mg NO₃ I⁻¹ and 0.05 mg PO₄I⁻¹ induced no significant alterations in bacterial mineralization of [¹⁴C]glucose (ANOVA; P >0.05) or incorporation of [³H]thymidine (ANOVA; P >0.05) in short-term (i.e. 3 h) experiments.
4. Microorganisms attached to refractory particulate organic matter do not appear to be limited by nitrogen or phosphorus even in streams with ambient nutrient concentrations as low as 0.06 mg NO₃ I⁻¹ and <0.03 mg PO₄ I⁻¹.
5. Our results indicate that variations in water temperature resulting from diurnal and seasonal temperature fluctuations, forest clear-cutting, and catchment elevation and aspect can have marked effects upon microbial activity and production, while short-term alterations in nutrient regime appear to have no significant effect on microbial activity associated with seston.     

Seasonal changes in the concentration and colour of humic substances in some aquatic environments

SUMMARY. 1. Between February and September 1974, concentrations of fulvic and humic acids in waters from a cryoboreal region of the Precambrian Shield in Quebec varied between 0.5–6.1 mg 1⁻¹ and 0.1–6.5 mg 1⁻¹ respectively. Whereas the fulvic acids usually remained relatively constant during this period, humic acids showed appreciable increases.
2. The number and weight average molecular weights of the fulvic acids (∼5000 and ∼22,000 daltons respectively) were lower than those of the humic acids (∼20,000 and ∼75,000 daltons respectively). The average molecular weight of the aquatic humic matter often decreased during late summer and early autumn. During spring floods the molecular weight of fulvic acids increased and that of humic acids declined.
3. Because the aquatic humic acids were more intensely coloured than fulvic acids, they contributed more than 30% of the colour of the water even though they accounted for only 15% of the concentration of humic matter. The colour intensity of both fulvic and humic acids generally appeared to increase during the latter part of the summer, when there was normally a decrease in the value of the colour quotient (E₄/E₆ ratio), indicating the accumulation in the waters of more humified material.     

Brain β-Spectrin Phosphorylation: Phosphate Analysis and Identification of Threonine-347 as a Heparin-Sensitive Protein Kinase Phosphorylation Site

Abstract: Phosphorylation of brain spectrin was studied by a combination of in vivo and in vitro approaches. Chemical analysis of phosphate groups on electrophoretically purified mouse brain β-spectrin yielded a stoichiometry of 3.2 ± 0.18 mol of PO₄/mol of β-spectrin. The spectrin isolated by chromatographic methods from mouse brain, pig brain, and human erythrocytes yielded 4.1, 5.6, and 3.2 mol of PO₄/mol of spectrin heterodimer, respectively. The ³²P labeling of spectrin in retinal ganglion cell neurons or NB 2a/d1 neuroblastoma cells with [³²P]orthophosphate showed phosphorylation of only β-spectrin in vivo. Two-dimensional phosphopeptide map analyses showed that most of the in vivo sites on β-spectrin were phosphorylated by either a heparin-sensitive endogenous cytoskeleton-associated protein kinase or protein kinase A. Phosphoamino acid analysis of in vivo and in vitro phosphorylated β-spectrin showed that [³²P]phosphate groups were incorporated into both serine (>90%) and threonine residues. In vitro, phosphate groups were incorporated into threonine residues by the heparin-sensitive endogenous protein kinase. The amino acid sequence VQQQLQAFNTY of an α-chymotryptic ³²P-labeled peptide phosphorylated by the heparin-sensitive cytoskeleton-associated endogenous protein kinase corresponded to amino acid residues 338–348 on the β1 repeat of β-spectrin_G (βSPIIa) gene. These data suggest that phosphorylation of Thr³⁴⁷, which is localized on the presumptive synapsin I binding domain of β-spectrin_G, may play a role in synaptic function by regulating the binding of spectrin to synaptic vesicles.     

Myelin P0 Glycoprotein: Identification of the Site Phosphorylated In Vitro and In Vivo by Endogenous Protein Kinases

Abstract: Myelin membrane prepared from mouse sciatic nerve possesses both kinase and substrates to incorporate [³²P]PO₄³⁻ from [γ-³²P]ATP into protein constituents. Among these, P₀ glycoprotein is the major phosphorylated species. To identify the phosphorylated sites, P₀ protein was in vitro phosphorylated, purified, and cleaved by CNBr. Two ³²P-phosphopeptides were isolated by HPLC. The exact localization of the sequences around the phosphorylated sites was determined. The comparison with rat P₀ sequence revealed, besides a Lys¹⁷² to Arg substitution, that in the first peptide, two serine residues (Ser¹⁷⁶ and Ser¹⁸¹) were phosphorylated, Ser¹⁷⁶ appearing to be modified subsequently to Ser¹⁸¹. In the second peptide, Ser¹⁹⁷, Ser¹⁹⁹, and Ser²⁰⁴ were phosphorylated. All these serines are clustered in the C-terminal region of P₀ protein. This in vitro study served as the basis for the identification of the in vivo phosphorylation sites of the C terminal region of P₀. We found that, in vivo, Ser¹⁸¹ and Ser¹⁷⁶ are not phosphorylated, whereas Ser¹⁹⁷, Ser¹⁹⁹, Ser²⁰⁴, Ser²⁰⁸, and Ser²¹⁴ are modified to various extents. Our results strongly suggest that the phosphorylation of these serine residues alters the secondary structure of this domain. Such a structural perturbation could play an important role in myelin compaction at the dense line level.     

Dissolved organic carbon concentrations and fluxes along the Moisie River, Quebec

SUMMARY. 1. The main stream and tributaries of a 145 km reach of the Moisie River, Quebec, were examined for temperature, conductivity, pH, dissolved organic carbon (DOC),%DOC>100,000 nominal molecular weight (NMW), optical density (OD₃₅₀), and the ratio of OD₄₀₀ to OD₆₀₀ (E4:E6).
2. Dissolved organic carbon concentrations correlated closely with OD₃₅₀ ( r ²=0.92, P <0.001). However,%DOC>100,000 NMW did not correlate with the E4:E6 ratio.
3. Except for a slight increase in%DOC> 100,000 NMW ( r ²=0.37, P <0.05), no change in any characteristic occurred down the length of the Moisie River, despite consistently higher levels of DOC in the tributaries.
4. Results suggest that high concentrations of DOC in tributary waters are rapidly removed within the main river channel. These results are discussed in terms of both biotic and abiotic models of in-strcam processing.     

Influence of dissolved humic materials on carbon assimilation and alkaline phosphatase activity in natural algal-bacterial assemblages

SUMMARY. Mixed natural assemblages of algae and bacteria exhibited lower rates of ¹⁴C assimilation and high rates of dissimilation of recent photosynthate when amended with low concentrations (7.2 mg 1^-1) of unfractioned dissolved humic materials (DHM). The extent of the inhibition or stimulation was greatest in the smaller (1–5 μm) assemblage particles. In different algal-bacterial assemblages, additions of DHM markedly enhanced community alkaline phosphatase activity (APA), particularly under low light regimes, DHM of low apparent molecular weight was much more stimulatory to both ¹⁴C assimilation and APA than DHM of high apparent molecular weight, supporting the belief that DHM molecular weight is an important determinant of DHM interactive capacity. Higher concentrations of D HM (either unfractionated, or molecular weight fractionated) produced greater APA responses. Addition of phosphate enhanced the disparity in rates of ¹⁴C assimilation of samples incubated under low and high light regimes, increased the rates of ¹⁴C assimilation, and depressed APA. There were indications of interactions between DHM and phosphorus in several experiments. Two hypotheses were invoked to explain increases in APA in response to DHM: (1) increased competition between algae and bacteria for phosphate following bacterial release from substrate limitation, or (2) DHM may have acted as a sequestering agent for organophosphorus compounds, and in so doing, gradually depleted available phosphate. In either case, it is clear that DHM alters phosphorus cycling. This DHM characteristic may be ecologically as important as its ability to complex trace metals.     

Grazing by oligochaetes and snails on epiphytes

SUMMARY. 1. The isotope ³³P was used to assess the effect of grazitig by oligochaetes (mainly Stylaria lacttstris L.) and the snail, Lymnaea peregra (Muller), on epiphytes within an Equisetum fluviatile L. stand.
2. Two 1 m² polystyrene enclosures were set up within the emergent macrophyte zone of the lake. At the start of the experiment ³³P-solution was mixed with the water in both tanks. Algal and animal samples for ³³P- analysis were collected during the peak occurrence of epiphytes in June.
3. Phosphorus release rates from the animals through defaecation and excretion were measured in the laboratory. The grazing rate of oligochaetes was 2.2–4.1 mg P m⁻² (of bottom) d⁻¹ of which about two - thirds was released and recycled. The oligochaete density ranged from 13,400 to 20,900 m⁻². The snails (25 ind. m⁻²) grazed 1.2-1.5 mg P m⁻²d⁻¹of whieh about a quarter was released through defaecation and excretion.
4. Daily consumption by the oligochaetes and snails corresponded to 22–45% of the average phosphorus uptake by the epiphytes.     

Phosphate transport in potato cuttings: Effect of gibberellic acid and abscisic acid

Apical cuttings of Solanum tuberosum L. cv. Sirtema were used al different stages of development to study long-distance transport of phosphate. The effects of two hormones, gibberellic acid (GA₃) and abscisic acid (ABA), on this process were also investigated. Before tuberization, phosphate (³²P) supplied to a single leaf was transported preferentially in the young and growing parts of the plant: apical bud, young leaves and roots. After tuberization, the tuber became the principal site of phosphate accumulation. GA³ treatment (10⁻⁴ M) of the tuber as well as of the leaves led to reduced transport of ³²P into the tuber. By contrast, treatment of the tuber with ABA (10⁻⁴M) did not change the ³²P distribution within the plant, while foliar spray with ABA greatly increased the transport into the tuber. The opposite effects of the two hormones on phosphate accumulation by tubers are discussed with regard to their opposite effects on the tuberization process.     

METHODS FOR THE STUDY OF BLOWFLY POPULATIONS

1. A simple method of radioactive labelling of L. sericata larvae is described.
2. More uniform count levels are obtained in individuals which have fed entirely on radioactive foods than in those which first feed on non-active food.
3. The radioactive count rates of the larvae are correlated with larval live weight and the counting rates of resulting imagines correlated both with corresponding larval counting rates and with imaginal live weight.
4. The loss of ³²P in the larvae due to causes other than decay of the isotope is traced from the prepupal stage to 3 weeks after emergence. The first and major loss occurs at emergence, when about 10% of the original ³¹P is shed as empty puparium and meconium. Adults lose about 1.5% per day.
5. The distribution of ³²P in the adult body of L. sericata labelled in the larval stage has been determined. The abdominal tissues have a lower activity than the rest of the body.     

Alteration of Tubulin-Gi Protein Interaction in Rat Cerebral Cortex with Aging

Abstract: The ability of the tubulin dimer to interact with and to modulate the G_i function inhibiting adenylyl cyclase was examined in cerebral cortex membranes from 2-month-old and 24-month-old rats. The hydrolysis-resistant GTP analogue 5'-guanylylimidodiphosphate (GppNHp)-dependent inhibition of adenylyl cyclase was significantly decreased in cerebral cortex membranes from 24-month-old rats. Tubulin, prepared from rat brains by polymerization with GppNHp, caused inhibition of adenylyl cyclase (∼28%) in 2-month-old rats. Tubulin-GppNHp-dependent inhibition of adenylyl cyclase in 24-month-old rats was significantly attenuated (∼15%). In 2-month-old rats, when tubulin, polymerized with the hydrolysis-resistant photoaffinity GTP analogue [³²P] P ³(4-azidoanilido)- P ¹-5'-GTP ([³²P]AAGTP), was incubated with cerebral cortex membranes, AAGTP was transferred from tubulin to G_iα. Transfer of AAGTP from tubulin to G_iα was reduced in 24-month-old rats. Furthermore, photoaffinity labeling of [³²P]AAGTP to G_iα in cortex membranes was significantly decreased in 24-month-old rats. No differences were observed in the amounts of G_sα, G_iα, or G_β subunits and tubulin, estimated by immunoblotting, in cortex membranes from 2-month-old and 24-month-old rats. These results suggest that the ability of tubulin to interact with G_i and thereby modulate the inhibitory regulation of adenylyl cyclase is reduced in the cerebral cortex of 24-month-old rats.     

Participation of Tubulin in the Stimulatory Regulation of Adenylyl Cyclase in Rat Cerebral Cortex Membranes

Abstract: This study examined effects of tubulin on the activation of adenylyl cyclase in rat cerebral cortex membranes. Tubulin, prepared from rat brain by polymerization with the hydrolysis-resistant GTP analogue 5'-guanylylimidodiphosphate (GppNHp) caused significant activation of the enzyme by ∼156% under conditions in which stimulation rather than inhibition of the enzyme was favored. Tubulin-GppNHp activated isoproterenol-sensitive adenylyl cyclase, potentiated forskolin-stimulated activity of the enzyme, and reduced agonist binding affinity for β-adrenergic receptors. When tubulin, polymerized with the hydrolysis-resistant photoaffinity GTP analogue [³²P] P ³(4-azidoanilido)- P ¹-5'-GTP ([³²P]AAGTP), was incubated with cerebral cortex membranes, AAGTP was transferred from tubulin to G_sα as well as G_iα. These results suggest that, in rat cerebral cortex membranes, the tubulin dimer participates in the stimulatory regulation of adenylyl cyclase by transferring guanine nucleotide to G_sα, as well as affecting the G_i-mediated inhibitory pathway.     

Reactions of singlet oxygen in humic waters

SUMMARY. The oxidation of 2, 5-diniethylfuran (DMF) to cis-1, 2- diacetylethylene (DAE) is a specific test for singlet oxygen (¹O₂). A method has been developed for the measurement of DAB by direct injection gas chromatography. By the use of this method, the photochemical generation of ¹O2 has been demonstrated in samples of two Canadian humic waters.
Two other photochemical reactions probably mediated by ¹O₂generation, the oxidation of histidine and the inactivation of a-chymotrypsin, have been demonstrated in these waters.
The possible ecological and environmental implications of these findings are discussed.     

Purification and biochemical characterization of a membrane-bound [NiFe]-hydrogenase from a hydrogen-oxidizing, lithotrophic bacterium, Hydrogenophaga sp. AH-24

Membrane-bound [NiFe]-hydrogenase from Hydrogenophaga sp. AH-24 was purified to homogeneity. The molecular weight was estimated as 100±10 kDa, consisting of two different subunits (62 and 37 kDa). The optimal pH values for H₂ oxidation and evolution were 8.0 and 4.0, respectively, and the activity ratio (H₂ oxidation/H₂ evolution) was 1.61 × 10² at pH 7.0. The optimal temperature was 75 °C. The enzyme was quite stable under air atmosphere (the half-life of activity was c . 48 h at 4 °C), which should be important to function in the aerobic habitat of the strain. The enzyme showed high thermal stability under anaerobic conditions, which retained full activity for over 5 h at 50 °C. The activity increased up to 2.5-fold during incubation at 50 °C under H₂. Using methylene blue as an electron acceptor, the kinetic constants of the purified membrane-bound homogenase (MBH) were V _max=336 U mg⁻¹, k _cat=560 s⁻¹, and k _cat/ K _m=2.24 × 10⁷ M⁻¹ s⁻¹. The MBH exhibited prominent electron paramagnetic resonance signals originating from [3Fe–4S]⁺ and [4Fe–4S]⁺ clusters. On the other hand, signals originating from Ni of the active center were very weak, as observed in other oxygen-stable hydrogenases from aerobic H₂-oxidizing bacteria. This is the first report of catalytic and biochemical characterization of the respiratory MBH from Hydrogenophaga .     

Identification of an Ectokinase Activity in Cerebellar Granule Primary Neuronal Cultures

Abstract: Primary neuronal cultures from 8-day-old rat cerebellum were incubated in the presence of exogenously added 16 n M [γ-³²P]ATP. Phosphorylation of a 45-kDa endogenous protein was detected within 1 min and increased linearly for ∼20 min. Unlike what was seen with [γ-³²P]ATP, in the presence of [³²P] orthophosphate no visible phosphorylation of protein was detected after 10 min, but a different pattern of phosphorylation was obtained in 30 min. The phosphorylation of the 45-kDa protein was reduced by 80–90% in the presence of 1 µ M unlabeled ATP, 5 U/ml of apyrase, or 0.01% trypsin but not 1 m M PO₄³⁻. Phosphorylation was inversely proportional to cell density and was unaffected by addition to the cells of 56 m M KCl or 100 µ M glutamate for 3 min. The presence of exogenously added cellular protein extracts or pretreatment of the cells for up to 20 min in phosphorylation buffer also did not affect the observed phosphorylation of the 45-kDa protein. The phosphorylation was found to be insensitive to MgCl₂ but inhibited in the presence of MnCl₂ or NaF and in the absence of CaCl₂. Analogues of ATP suppressed phosphorylation of the 45-kDa protein by 80–90%. A similar inhibition was obtained in the presence of ADP or AMP. In this study, we establish via several different means that the phosphorylation of the 45-kDa protein in primary neuronal granule cultures occurs extracellularly through an ectokinase activity, which is furthermore distinguishable from a series of other presently characterized ecto-protein enzymes and intracellular kinases.     

Polypeptide Elongation Factors of the Developing Chick Brain

Abstract: The polypeptide elongation factors (EF-1_L, EF-1_H, and EF-2) of the developing chick brain were separated and purified by means of a combination of gel chromatographic methods. The molecular weight of EF-1_H of the chick brain ranged from 5 to 10 × 10⁵, and was different from that of the chick liver (about 7 × 10⁵). The molecular weight of other purified factors was about 5 × 10⁴ for EF-1_L. and 9.4 × 10⁴ for EF-2. High activities of polyphenylalanine (poly-Phe) synthesis per mg protein in the developing chick brain were observed between the 3rd embryonic week and the 1st post-hatch week and declined afterwards. On the other hand, the levels of both EF-1 and EF-2 per mg protein in the brain were observed to be high in an early embryonic stage, gradually declining afterwards to the adult level. The brain EF-1_L was a major component of EF-1 in an early embryonic stage, while EF-1_H became recognizable in the 3rd embryonic week. Moreover, the EF-1_H activities were found to be more than double with regard to the binding reaction and to be more than 10-fold as active in respect to poly-Phe synthesis in comparison with the activities of EF-1_L. It is proposed that the brain EF-1_H could be due to aggregates consisting of EF-1_L, a stimulatory factor, and other components.     

Microbial transformation of labile dissolved organic matter into humic-like matter in seawater

Abstract Microbial transformation of labile, low molecular weight dissolved organic matter (DOM) into dissolved humic matter (DHM) was studied in seawater. Surface water samples were amended with [¹⁴C into ¹⁴CO₂, TO¹⁴C (total organic ¹⁴C), and PO¹⁴C (particulate organic ¹⁴C), was measured over time in confined samples. The humic and non-humic fractions of DO¹⁴C (dissolved organic ¹⁴C) were separated according to a common operational definition of DHM based on adsorption on XAD-8 macroporous resin. Both TO¹⁴C and non-humic DO¹⁴C decreased during the experiments. However, ¹⁴C-labelled DHM increased during the first week of the incubations, to a level where it comprised 15% of the TO¹⁴C remaining in the samples, or 3% of the initially added ¹⁴C. Towards the end of experiments (ca 70 days), the humic fraction of DO¹⁴C gradually approached the background level of poisoned control samples. Provided that the XAD-8 operational definition of DHM is accepted, this study indicates that humic matter may be formed in seawater within days from labile monomers such as glucose.     

Ecology of Peridinium willei and P. volzii (Dinophyceae) in Danish lakes

The distribution of Peridinium willei and P. volzii was studied in Danish lakes. Both species were confined to lakes with concentrations of Total P < 0.15 mg 1^-1, with the majority of occurrences at Total P concentration between 0.020–0.040 mg 1^-1 and concentrations of PO₄ P between detection limit and 0.040 mg 1^-1. The occurrence of the species in relation to inorganic N compounds (NH₄ N and NO₂+ NO₃ N) was significantly broader for P. willei than for P. volzii: P. willei had an almost even distribution within a wide range of NH₄ N, whereas P. volzii mainly occurred between 0.001 and 0.10 NH₄ N 1^-1. P. willei had an almost even distribution at values beween 0.005 and 0.42 mg NO₂+ NO₃ N 1^-1, whereas P. volzii mainly occurred below 0.050 mg NO₂+ NO₃ N 1¹. P. willei was found at pH values between 4.2 and 8.5, whereas P. volzii was confined to lakes with a slightly basic pH. The study confirmed the broad limits of P. willei and the much more narrow limits of P. volzii in relation to seasonal occurrence and pH, as well as an affinity of the former to ponds and lakes with a rich bottom vegetation. The study also showed, however, that the species were not as widespread and common in recent Danish lake phytoplankton as generally stated by previous authors. The use of different ecological factors to give weight to species separation is discussed. The inclusion of P. volzii in P. willei proposed by Popovsky & Phiester is not supported by the present study, as the two taxa appear to have different ecological tolerances.     

UPTAKE OF [14C]ASPARTIC ACID AND [14C]GLUTAMIC ACID BY RETINAL SYNAPTOSOMAL FRACTIONS

Abstract— Uptake systems for [¹⁴C]aspartate and [¹⁴C]glutamate were characterized in two distinct synaptosomal fractions solated from rabbit retina. The P, synaptosomal fraction was highly enriched in large photoreceptor cell synaptosomes but contained very few conventional sized synaptosomes from amacrine, horizontal or bipolar cells. In contrast, the P₂ synaptosomal fraction contained numerous conventional sized synaptosomes and was virtually free of photoreceptor cell synaptosomes. Both synaptosomal fractions took up [¹⁴C]aspartate and [¹⁴C]glutamate with high affinity [ K _m= 1–2μM). Uptake characteristics were similar to those described for high affinity uptake systems in brain synaptosomes, i.e. saturation kinetics; temperature and Na⁺ dependence. Although the presence of a high affinity uptake system is not a definitive criterion for demonstration of functional neurotransmitter systems, it is an important and necessary prerequisite and can thus be considered as supportive evidence for the involvement of asparate and glutamate in neurotransmission in rabbit retina.     

31P NMR Relaxation Does Not Affect the Quantitation of Changes in Phosphocreatine, Inorganic Phosphate, and ATP Measured In Vivo During Complete Ischemia in Swine Brain

Abstract: Ischemia-induced changes in ³¹P NMR relaxation were examined in 16 piglets. NMR spectra were acquired under control conditions and during complete cerebral ischemia induced via cardiac arrest. Changes in T ₁ were assessed directly in six animals during control conditions and after 30–45 min of complete ischemia when changes in brain P₁ levels had reached a plateau. The T ₁ for P₁ did not change, i.e., 2.3 ± 0.5 s during control conditions versus 2.4 ± 1.0 s during ischemia. To evaluate phosphocreatine and ATP, two types of spectra, with a long (25-s) or short (1-s) interpulse delay time, were collected during the first 10 min of ischemia (n = 10). Both types of spectra showed the same time course of changes in phosphocreatine and ATP levels, implying that the T ₁ relaxation times do not change during ischemia. There were no changes in the linewidths of phosphocreatine, ATP, or P₁ during ischemia, implying that the T *₂ values remain constant. Our results suggest that the ³¹P T ₁ and T *₂ for phosphocreatine, P_i, and ATP do not change during ischemia, and therefore changes in ³¹P NMR peak intensity accurately reflect changes in metabolite concentrations.