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Biomedical subjects

B Chance

Publications and source records attributed to B Chance.

At least 289 records · Page 16Linked to original sources
PubMed1986

Cytochrome c peroxidase compound ES is identical with horseradish peroxide compound I in iron-ligand distances.

X-ray absorption studies of compound ES of cytochrome c peroxidase show a short iron-oxygen distance of 1.67 +/- 0.04 A, an iron-histamine distance of 1.91 +/- 0.03 A, and an iron-pyrrole nitrogen average distance of 2.02 +/- 0.02 A. This is identical within the error with the reported structure of horseradish peroxidase compound I [Chance, B., Powers, L., Ching, Y., Poulos, T., Yamazaki, I., & Paul, K. G. (1984) Arch. Biochem. Biophys. 235, 596-611]. Comparisons of the structures of myoglobin peroxide [Chance, M., Powers, L., Kumar, C., & Chance, B. (1986) Biochemistry (preceding paper in this issue)], compound ES, and the intermediates of horseradish peroxidase reveal the possible mechanisms for the stabilization of the free radical species generated during catalysis. The proximal histidine regulates the structure and function of the pyrrole nitrogens and the heme, allowing for the formation and maintenance of the characteristic intermediates.

Binding Sites↗
PubMed1986

Three-dimensional metabolic mapping of the freeze-trapped brain: effects of ischemia in the Mongolian gerbil.

The effects of unilateral carotid artery (left or right) occlusion on the three-dimensional metabolic mapping were studied in the Mongolian gerbil (Meriones unguiculatus). The brain was freeze-trapped by the liquid nitrogen funnel technique and was analyzed for the two-dimensional distribution of the reduced pyridine nucleotides and oxidized flavoprotein using a time-sharing micro-light guide surface fluorometer/reflectometer. The results could be summarized as follows. Bilateral carotid artery occlusion induced uniform effects in terms of redox state in the cerebral cortex and the olfactory bulbs. After unilateral carotid occlusion, the redox state of the ischemic ipsilateral hemisphere was higher and was not affected by blood volume changes as evaluated from the reflectance signal scanned in several brains. The narrow band of tissue near the midline of the ischemic hemisphere connected to the contralateral hemisphere blood supply appeared in a large number of gerbils. The anomaly in the redox state of the intracellular space also appeared in subcortical structures, as seen in all the two-dimensional mappings measured. It seems that blood circulation to the two olfactory bulbs is connected to the same blood vessel. It is suggested that unilateral carotid artery-occluded gerbils may show large variability inside the same hemisphere and therefore results must be evaluated very carefully.

Animals↗
PubMed1986

Uncoupling effects of 2,4-dinitrophenol on electron transfer reactions and cell bioenergetics in rat brain in situ.

We have examined the effects of uncoupling on electron transfer reactions and cell bioenergetics in rat brain in situ. The redox reactions of cyt aa3, cyt c and cyt b as well as the levels of phosphocreatine/inorganic phosphate ratio (PCr/Pi) were monitored in isolated perfused rat head, using optical and [31P]NMR techniques, respectively. In the first series of experiments anesthetized (pentobarbital) and mechanically ventilated rats underwent bilateral carotid arterial cannulation. The head (skull intact, muscle removed) was perfused with a perfluorochemical solution (FC-43), and the animal was then decapitated. By means of reflectance spectrophotometry, the redox reactions of cyt aa3, c and b were monitored before and after perfusing the head with uncoupler 2,4-dinitrophenol (DNP) as well as after complete anoxia. The second series of experiments was performed using [31P]NMR spectroscopy (24.33 MHz; 10.25 in. magnet). PCr/Pi ratios were first determined for living rats (in vivo) before and after exchange transfusion with FC-43 (Hct less than 0.5%), and then in isolated perfused head (in situ) before and after 2,4-DNP addition. Reduction of cyt aa3 (32 +/- 1.4%, mean +/- S.E.M.), cyt c (29 +/- 6%), and cyt b (19 +/- 2%) was induced by addition of 2,4-DNP. Compared to in vivo aerobic metabolism with a PCr/Pi ratio of 3.0-5.0, the value in the uncoupled state in situ was less than 0.1. We conclude that in contrast to in vitro, uncoupling of oxidative phosphorylation induces reduction of intramitochondrial cytochromes in situ.(ABSTRACT TRUNCATED AT 250 WORDS)

2,4-Dinitrophenol↗
PubMed1986

Magnetic resonance spectroscopy of normal and diseased muscles.

Phosphorus magnetic resonance spectroscopy (P MRS) affords and innovative approach to the study of the oxidative enzyme content of normal and diseased muscles. Examples of the evaluation of the enzyme content of normal muscles by an exercise protocol are provided. The protocol affords a hyperbolic work/cost profile, the Vmax of which is calculated by the reciprocal plots giving the enzyme content and the "effective Michaelis constant" with an evaluation of the resting metabolism. This steady state protocol clearly illustrates enzyme adaptation, on the one hand, and tissue atrophy particularly in the case of tissue injury, Duchenne's dystrophy, and genetic deletion of specific enzymes, on the other hand. The method is rapid, safe, and affords a quantitative evaluation of the disease process and possibilities for following appropriate therapies. So far, approx 1000 examinations of normal and diseased human limbs have been carried out in our laboratory in over the past four years.

Acidosis↗
PubMed1986

Treatment of mitochondrial myopathy due to complex III deficiency with vitamins K3 and C: A 31P-NMR follow-up study.

A patient with mitochondrial myopathy due to complex III deficiency who was treated with vitamin K3 (menadiol sodium diphosphate, 40 mg daily) and vitamin C showed clinical improvement. A 1-year study with phosphorus 31 nuclear magnetic resonance (31P-NMR) monitoring has shown that clinical and metabolic improvement was maintained by this therapy; increasing the dose of vitamin K3 to 80 mg daily improved the bioenergetic state of the patient's muscles at rest; postexercise recovery was less responsive to the increased dose; and a higher dose of vitamin K3 (80 mg/day) did not produce side effects. The differential therapeutic effects of vitamin K3 at rest and during exercise recovery are probably due to the differential kinetics of each metabolic state. Monitoring muscle bioenergetics with 31P-NMR is valuable in documenting therapeutic improvements in mitochondrial myopathies.

Adult↗
PubMed1986

Cerebral metabolic effects of neonatal seizures measured with in vivo 31P NMR spectroscopy.

In vivo phosphorus 31 nuclear magnetic resonance (31P NMR) spectroscopy was used to evaluate changes in cerebral high-energy phosphate compounds in 8 infants with seizures. During the study 4 babies had seizures that caused a 50% decrease in the phosphocreatine to inorganic phosphate (PCr/Pi) ratio. Focal seizures caused lateralized decreases in the PCr/Pi ratio; generalized seizures caused bilateral decreases. Postictal spectra had increased PCr/Pi ratios, presumably due to postictal inhibition. Interictal 31P NMR spectra were normal. One patient's seizures were successfully treated with intravenously administered phenobarbital during NMR data acquisition, causing an immediate increase in the PCr/Pi ratio from 0.7 to 1.2. These studies indicate that cerebral PCr concentration decreases by approximately 33% and that oxidative metabolism increases by approximately 45% during neonatal seizures. Five babies had PCr/Pi ratios of less than 0.8 during seizures and subsequently developed long-term neurological sequelae, which suggests that neonatal seizures may cause or exacerbate cerebral injury by increasing cerebral metabolic demands above energy supply.

Adenosine Triphosphate↗
PubMed1986

Redox studies of the epiphyseal growth cartilage: pyridine nucleotide metabolism and the development of mineralization.

The objective of this investigation was to examine the redox status of chondrocytes in normal and rachitic growth cartilages and to relate energy metabolism to cell maturation and the initiation of mineralization. The redox status was evaluated by chemical analysis and by microfluorimetric scanning of rapidly frozen, freeze-fractured tibial growth cartilages. In the normal epiphysis, the redox pattern of both avian and lagomorph cartilages were very similar. Thus, in the proliferative tissue zone the NAD/NADH ratio was high; in the hypertrophic zone, the cells appeared to be reduced. The sharp border between the two zones suggested that the redox shift may be associated with development of hypoxia. Induction of rickets resulted in a fivefold decrease in the total concentration of pyridine nucleotides in the proliferating and hypertrophic zones. Furthermore, the NAD/NADH ratio was profoundly disturbed. In the mineralizing zone, there was an accumulation of reduced pyridine nucleotide. Healing, initiated by administration of vitamin D to the rachitic birds, caused a rapid increase in NAD and NADH in all zones of the growth cartilage. It was concluded that vitamin D deficiency leads to changes in the energy metabolism of growth cartilage and that these changes were related to the defective mineralization of the rachitic tissue.

Animals↗
PubMed1986

Electrical stimulation of human muscle studied using 31P-nuclear magnetic resonance spectroscopy.

This study used phosphorous nuclear magnetic resonance (31P-NMR) spectroscopy to examine the metabolic demand resulting from electrical muscle stimulation (EMS) applied to human skeletal muscle. For each of six subjects, the forearm flexor muscle group was monitored with 31P-NMR during both maximum voluntary and 6-s EMS-induced contractions. A simple protocol using a tourniquet was added in one subject to assess the role of blood flow in this model. Eight hertz (nontetanic) EMS showed less (p less than 0.025) depletion of phosphocreatine (36%) than did tetanic 70-Hz EMS (60%), voluntary isometric (66%), and voluntary isokinetic (68%). The results of the tourniquet studies suggested that the nontetanic EMS allowed relatively increased muscle blood flow and oxygen supply during contraction. Tetanic EMS provided a similar metabolic demand to that of conventional resistive exercise, as measured by 31P-NMR spectroscopy.

Adult↗
PubMed1986

31P NMR detection of mobile dog brain phospholipids.

The in vivo dog brain 31P NMR spectrum has a large peak in the phosphodiester region accounting for more than 35% of the total observable phosphorus metabolites. It is possible to reduce the intensity of this peak by off-resonance saturation. To characterize the nature of this peak, extracts of dog brain frozen in situ were analyzed by high resolution 31P NMR. ATP, phosphocreatine, inorganic phosphate, and phosphomonoesters were recovered in appropriate amounts in the methanol:HCl extract. However, acid soluble phosphodiesters accounted for only 8% of the observable phosphorus. More NMR observable phosphodiesters were selectively recovered following CHCl3:methanol extraction of phospholipids. These results suggest that the in vivo phosphodiester resonance has substantial contributions from a fraction of mobile brain phospholipids.

Animals↗
PubMed1986

Myocardial high energy phosphate metabolism in closed chest dog: creation of an animal model.

A chronic closed chest dog model was developed to study myocardial metabolism with NMR spectroscopy. Cardiac windows were surgically created in 10 dogs by removal of two ribs and accompanying skeletal muscle. Marlex mesh was sewn between the two exposed ribs, and fascia and skin were closed. 31P spectra were obtained using a surface coil placed into the surgically created pouch and using routine NMR pulsing techniques.

Adenosine Triphosphate↗
PubMed1986

P NMR evaluation of hypoxic stress in brain of animal models.

Quantification by NMR of hypoxic stress is best afforded by the ratio of phosphocreatine (PCr) to inorganic phosphate (Pi). The thin cranium and sternum of neonates affords ideal conditions for NMR evaluation of impaired oxidative metabolism. Brains of animal models (dog and cat) are best studied with muscles retracted with the rf coil placed on the bare skull. 31P and 1H (lactate) signals are time shared with a doubly tuned coil. Servo-stabilized hypoxia gives simplified steady state analysis. Hypoxic insults of the brain are evaluated by the integral of the deviation of the PCr/Pi ratio from the normoxic values. Generally, a deviation of 1 unit of PCr/Pi for an interval of 3 hr will lead to a severe metabolic damage and recovery from hypoxia is often 30-fold delayed. Comprehensive monitoring of energy metabolism (PCr/Pi), respiratory chain redox states, intracellular pH and lactate accumulation allow the calculation of integrated pH and lactate "insults" and thus offer new insights on the effects of hypoxia on brain oxidative metabolism.

Animals↗
PubMed1986

Relaxation and imaging of lithium in vivo.

It has recently been demonstrated that NMR imaging can be used to record the distribution of lithium. We now report on the parameters pertinent to this imaging. Specifically, the relaxation of Li in aqueous solution, in agarose gel, and in vivo has been investigated. In the latter case, both the longitudinal and transverse relaxations were biexponential, consistent with the behavior expected for a spin 3/2 quadrupole relaxed nucleus. The overall relaxation rate was quite slow in vivo with T'1 = 3.5 sec and T''1 = 6.6 sec.

Animals↗
PubMed1986

Detection of skeletal muscle hypoperfusion during exercise using phosphorus-31 nuclear magnetic resonance spectroscopy.

Blood flow to working skeletal muscle is frequently reduced in patients with heart failure or peripheral vascular disease. To determine if phosphorus nuclear magnetic resonance (NMR) can noninvasively detect such muscle underperfusion, gated phosphorus-31 NMR spectroscopy was used to compare muscle inorganic phosphate, phosphocreatine and pH during mild wrist flexion exercise at 0.2, 0.4 and 0.6 W in eight normal men, before and after reduction of forearm blood flow. Forearm flow was reduced by cuff inflation to a pressure determined by Doppler ultrasound to bring flow to 40 to 60% of control. Attention was focused on the inorganic phosphate to phosphocreatine (Pi/PCr) ratio and pH, two variables potentially sensitive to muscle oxygen delivery. At rest with normal flow, Pi/PCr averaged 0.12 +/- 0.03 and pH averaged 7.02 +/- 0.04. Exercise produced a progressive increase in Pi/PCr (0.2 W = 0.43 +/- 0.12; 0.4 W = 0.75 +/- 0.31; 0.6 W = 1.04 +/- 0.47) and a modest decrease in pH (0.2 W = 6.94 +/- 0.04; 0.4 W = 6.86 +/- 0.18; 0.6 W = 6.85 +/- 0.06). Flow reduction had no effect on Pi/PCr or pH at rest. In contrast, flow reduction during exercise was associated with higher Pi/PCr at all three work loads (0.2 W = 0.60 +/- 0.27; 0.4 W = 0.99 +/- 0.50; 0.6 W = 2.00 +/- 1.26 [all p less than 0.05 versus normal flow]) and lower pH (0.2 W = 6.78 +/- 0.12; 0.4 W = 6.69 +/- 0.23; 0.6 W = 6.65 +/- 0.30 [p less than 0.01 versus normal flow at 0.2 and 0.4 W; p = 0.05 at 0.6 W]).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1986

31P nuclear magnetic resonance spectroscopy: noninvasive biochemical analysis of the ischemic extremity.

The biochemical effects of peripheral vascular disease on skeletal muscle have not been characterized precisely because of the lack of satisfactory noninvasive analytic methods. 31P nuclear magnetic resonance (NMR) spectroscopy was used to measure the high-energy phosphate compounds, phosphocreatine (PCr) and adenosine triphosphate, as well as metabolic byproducts, such as inorganic phosphates (Pi) and phosphate monoesters in calf muscles of 214 limbs with peripheral vascular disease. Intracellular pH was also measured. The NMR index (Pi[PCr + Pi]) was used to quantitate the impairment of oxidative phosphorylation as a result of ischemia. Studies done at rest documented the impairment of oxidative metabolism only in limbs with severe ischemia (ankle-brachial pressure index (API) less than 0.4). Exercise resulted in a significant elevation of the NMR index in all limbs and the rate of return of this value toward normal following exercise was prolonged even in limbs with moderate ischemia (0.4 less than or equal to API less than or equal to 0.9). Correlation of 31P NMR parameters with arteriograms showed that infrapopliteal occlusions resulted in prolonged recovery times only when the superficial femoral artery was occluded and emphasized the metabolic consequences of multisegmental disease. Accumulation of glycolytic pathway intermediates correlated with the decrease in muscle cell pH observed with exercise. Despite immediate improvement in symptoms and hemodynamic parameters following revascularization, return to normal biochemical function occurs over a prolonged period of time. This study demonstrates that 31P NMR spectroscopy can successfully measure noninvasively the important phosphorus-containing compounds involved in the bioenergetics of skeletal muscle in vivo rapidly enough to permit real-time determination during exercise and recovery.

Adenosine Triphosphatases↗
PubMed1986

Multiple controls of oxidative metabolism in living tissues as studied by phosphorus magnetic resonance.

Three types of metabolic control of oxidative metabolism are observed in the various tissues that have been studied by phosphorous magnetic resonance spectroscopy. The principal control of oxidative metabolism in skeletal muscle is by ADP (or Pi/phosphocreatine). This conclusion is based upon studies of arm muscles of humans during steady-state exercise. A work-cost (Vm vs. Pi/phosphocreatine) relationship follows a Michaelis-Menten rectangular hyperbola, where Km values from 0.5 to 0.6 and Vmax values from 50 to 200 (at nearly constant pH) are found in linearized plots of the equation V/Vmax = 1/(1 + 0.6 phosphocreatine/Pi) where V is work level (which is equal to the velocity of the enzymatic reaction) and Vmax is the maximal work capacity that is a measure of the enzyme activity (E) of oxidative metabolism. Adaptation to exercise enhances the slope of the work-cost relationship and causes large changes in Vmax or E. A second metabolic control may enhance the slope of the work-cost relationship but not Vmax. For example, the initiation of exercise can lead to an improved characteristic that can be explained by 2-fold increased substrate delivery, for example, increased oxygen delivery by microcirculatory control. Cardiac tissue of the adult dog affords an example of optimal endurance performance adaptation and exhibits the steepest work-cost relationship observed and is attributed to a coordinated control of substrate delivery that may involve Ca2+ and inorganic phosphate control of NADH; control of O2 delivery may also be involved. The calculated work-cost relationship is similar to that observed in the beagle heart. The theoretical curve illustrates that the liability of multiple controls is a sharp break point in metabolic control at the end of the multiple control range--a possible cause of instability of cardiac performance at high V/Vmax.

Acidosis↗
PubMed1986

Acute intestinal ischemia studies by phosphorus nuclear magnetic resonance spectroscopy.

31P nuclear magnetic resonance (NMR) spectroscopy has been used to follow the metabolism of acutely ischemic rat small intestine and its recovery after reversal of ischemia. Loops of small intestine were subjected to occlusive external pressure for up to 60 minutes, followed by a recovery period. The depletion of PCr and ATP is rapid and complete within 20 minutes. Recovery from ischemia is also rapid but with recovery ATP levels lower than initial values after prolonged ischemic periods. Intestinal shock was avoided. Clinical recovery correlated with shorter ischemic periods. 31P NMR spectroscopy thus appears to be a suitable technique for studying the effects of pharmacological agents and other treatments for amelioration of ischemic effects on the bowel.

Adenine Nucleotides↗
PubMed1986

Phosphorus magnetic resonance spectroscopy studies of the role of mitochondria in the disease process.

The incisive detection of bioenergetic insufficiency in an organ of known workload by P MRS is noninvasive and nondestructive, and in some cases the portion of the organ involved can be determined, particularly if both PCr and ATP are depleted. The fractional loss of ATP and hence the relative volumes of viable and "metabolically dead" tissue are thereby evaluated. In addition, the value of P MRS in following a therapy complements its value in diagnosis as this has been demonstrated in cases followed over 6 months to three years. The fact that deficiencies of the enzymes and substrates of oxidative metabolism can be detected by P MRS affords a global overview of energy metabolism that can be a key to rapid diagnosis. The distinction of the enzyme and/or substrate deficiency, while not directly indicated by steady state P MRS, can be identified by use of the "Crossover Theorem" and its impact upon blood and tissue levels of substrates (including oxygen). In the case of neonatal systemic hypoxia, there is no doubt about which of the equations applies, and similarly in metabolic disease, a glutaric acid urea is a direct consequence of the crossover response of metabolism and signifies that an enzyme deficiency may be involved. Furthermore, the clinical danger of a high Pi/PCr value is clarified by our observations, both from the animal models and from the theory, the high clues; i.e. 2 and over, suggest work stresses near the capability of oxidative metabolism and imminent failure of the negative feedback afforded by metabolic regulation, particularly ADP control of oxidative metabolism. This control is lost because of the fall of phosphocreatine to the point where creatine kinase is no longer in equilibrium, leading to the loss of ATP and its conversion to large amounts of ADP and its breakdown products. ATP then stimulates glycolysis and results in a massive lactic acidosis. At the same time, the low thermodynamic capability of glycolytic metabolism is unable to prevent irreversible ion disequilibration, water movements, edema, and eventually rupture of the cell membrane. The pathway of resynthesis of ATP is then tortuous, particularly as AMP is deaminated and adenosine is converted eventually to hypoxanthine. Thus, NMR reports that metabolic control is operating in the region where homeostasis of biochemical parameters is feasible. It further reports regions where the metabolic control is susceptible to failure and most aggressive clinical care is required.

Adenosine Triphosphate↗