Biomedical subjects

W J Ray

Publications and source records attributed to W J Ray.

At least 55 records · Page 3Linked to original sources

PubMed1985

EEG activity during cognitive processing: influence of attentional factors.

The research draws upon two separate areas of inquiry: that of lateralized electrocorical processing of cognitive material; and that of psychophysiological information processing studies related to foci of attentional demands. Eighteen subjects on each of three separate days were presented with tasks considered to be 'right hemispheric' or 'left hemispheric'. These tasks were paired in a 2 X 2 design with an attentional factor requiring attention to the environment or to internal processing. All subjects received all types of tasks. Bilateral EEG measures were taken from the frontal and parietal areas referenced to linked ears. Major frequency bands were determined by a Fast Fourier Transform (FFT) and appropriate ANOVAs performed. The results suggest that task factors (left vs right hemisphere tasks) and attentional demands (internal vs external) are differentially represented in terms of EEG functioning. In general the higher beta frequencies were more sensitive to the hemispheric tasks demands and the middle frequencies (alpha and low beta) more sensitive to the attentional demands especially in the parietal areas.

Alpha Rhythm↗

PubMed1984

Acceleration of the NAD cyanide adduct reaction by lactate dehydrogenase: the equilibrium binding effect as a measure of the activation of bound NAD.

The binary complex of NAD and lactate dehydrogenase reacts reversibly with cyanide to produce a complex (E X NAD-CN) whose noncovalent interactions are similar to those in the E X NADH complex (where E is one-fourth of the tetrameric dehydrogenase). The reaction apparently is a simple bimolecular nucleophilic addition at the 4 position of the bound nicotinamide ring; viz., cyanide does not bind to the enzyme prior to reaction. The value of the dissociation constant for E X NAD-CN is about 1 X 10(-6) M and is independent of pH over the range of 6-8. The equilibrium constant for the reaction of cyanide with E X NAD is about 400-fold larger than that for the nonenzymic process after a statistical correction. This increment in Ke is accounted for by a 220-fold increase in the rate of the forward enzymic reaction (20 M-1 s-1) as compared with an approximately 2-fold decrease for the reverse process (9 X 10(-5) s-1). Thus, the increased value of the rate constant for bond formation in the enzymic reaction is attributed to an equilibrium binding effect that is translated almost entirely into a rate effect on that step (bond formation). Since the nonenzymic reaction is sensitive to solvent composition, this equilibrium binding effect likely is produced by environmental effects at the nicotinamide/dehydronicotinamide part of the coenzyme binding site on the enzyme.

Animals↗

PubMed1984

The lactate dehydrogenase catalyzed pyruvate adduct reaction: simultaneous general acid-base catalysis involving an enzyme and an external catalyst.

The pH dependence of the reaction catalyzed by lactate dehydrogenase, where pyruvate adds covalently to NAD to yield a NAD-Pyr adduct, together with published data on the pH dependence of parameters in the normal redox reaction suggests similar binding modes for enolpyruvate and lactate in their complexes with E X NAD (where E is one-fourth of the tetramer), for ketopyruvate in its complexes with the protonated species, E X H X NAD and E X H X NADH, and for the NAD--Pyr adduct and NADH plus pyruvate in their complexes with E X H. These similarities, together with previous data, suggest a reaction scheme for the formation of the enzyme-adduct complex that includes the relevant proton-transfer steps. Seven different amine chloride buffers were used in a study of the reverse adduct reaction, i.e., the decomposition of E X H X NAD--Pyr. These act with varying efficiencies as external general acid catalysts; the enzyme apparently acts as a (internal) general base. The involvement of the amine chloride buffers as external general catalysts is supported by the concentration dependence of the buffer effect, by a Brönsted plot, and by solvent deuterium isotope effects. The involvement of the enzyme as an internal general catalyst is inferred from the pH dependence of the reaction and the identities of the nearby groups in the E X H X NAD--Pyr complex (from crystallographic studies). The dependence of the adduct reaction on chloride concentration indicates the presence of dead-end inhibitor complexes of E X H X Cl and E X H X NAD X Cl. Chloride also accelerates the decomposition of the adduct in the complex E X H X NAD--Pyr by binding to this complex.

Animals↗

PubMed1984

On the origin of the lactate dehydrogenase induced rate effect.

To evaluate the ability of lactate dehydrogenase to facilitate the bond making/breaking steps for both the addition of pyruvate enol to NAD (pyruvate adduct reaction) and the normal redox reaction, the ability of the enzyme to facilitate the tautomerization of bound pyruvate is assessed. In addition, the equilibrium constants for the adduct reaction are obtained for both bound and free reactants from the ratio of the rate constants in the forward and reverse reactions (at pH 7). The latter comparison indicates that the enzyme facilitates bond making/breaking in the (forward) pyruvate adduct reaction by a factor of about 10(11) M. Similar comparisons suggest that reactant immobilization accounts for about 1000 M of this 10(11) M rate effect. Since the (pH-independent) rate constant for the ketonization of bound pyruvate enol assisted by the external buffer, imidazolium ion, is 2 X 10(7) M-1 s-1 and the corresponding rate constant for free pyruvate enol, again assisted by imidazolium ion, is 35 M-1 s-1 [Burger, J. W., II, & Ray, W. J., Jr. (1978) Biochemistry 17, 1664], the enzyme facilitates the bond making/breaking steps associated with the conversion of bound HO-C less than to bound O = C less than by a factor of about 10(6)-fold. The product of the above two rate enhancement factors and the rate factor suggested previously for the environmental effect on NAD produced by its binding to lactate dehydrogenase, 100-fold, is 10(11) M, and it accounts for the bond making/breaking effects exerted by the enzyme in the pyruvate adduct reaction. The rate constant for oxidation of ethanol (a model for lactate) by 1-methylnicotinamide (a model for NAD) is about 5 X 10(-12) M-1 s-1 at 25 degrees C in pure ethanol (delta H for this reaction is about 30 kcal/mol). The ratio of the rate constants for E X NAD X Lac----E X NADH X Pyr and the above model reaction is estimated as about 10(14) M in water; i.e., the LDH-induced rate effect is about 10(14) M. The product of the values for the above rate factors for the normal redox reaction is about 10(12) M. Although the value of this product is less certain than that for the adduct reaction, these rate factors do account for much of the LDH-induced rate effect.

Animals↗

PubMed1984

Enzyme-bound intermediates in the conversion of glucose 1-phosphate to glucose 6-phosphate by phosphoglucomutase. Phosphorus NMR studies.

The interactions between metal ions and the phospho form of rabbit muscle phosphoglucomutase (EC 2.7.5.1) have been studied by 31P NMR. In the metal-free enzyme, the width at half-height of the 31P signal is 10 +/- 1 Hz at 81 MHz. In enzyme-Cd2+ complexes, the presence of spin-spin coupling with 113Cd2+ (J113Cd-O-31P = 16 Hz) and the absence of such splitting with 114Cd2+ indicate that Cd2+ binds directly to the enzymic phosphate. The absence of detectable splitting on transfer of the phosphate group to the acceptor hydroxyl group of bound glucose 1-phosphate, or glucose 6-phosphate (to give the 113Cd2+ complex of the dephospho-enzyme and glucose 1,6-bisphosphate), indicates that this transfer eliminates the direct metal ion-phosphate interaction. The enzyme-catalyzed reaction is slowed sufficiently by the addition of Li+ to allow studies of three discrete intermediate complexes by NMR techniques: glucose 1-phosphate bound to the phosphoenzyme, glucose 1,6-bisphosphate bound to the dephosphoenzyme (only one complex of this type was observed), and glucose 6-phosphate bound to the phosphoenzyme. Complete assignments of the phosphorus resonances of these intermediates have been made by labeling the phosphate ester group of either the enzyme or the sugar with 17O and by NMR polarization transfer studies. The effect of bound metal ions on these resonances also was determined. A 31P NMR titration study of the Li+ complex of the dephosphoenzyme with glucose 1,6-bisphosphate and a 31P NMR polarization transfer experiment indicate that beta-glucose 1,6-bisphosphate binds to the enzyme less tightly than alpha-glucose 1,6-bisphosphate.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1984

Self-report and heart rate responses to a stressful task.

Data were obtained from a risk task, the zipwire, in order to examine self-report and heart rate responses of males and females. Using this task, heart rate data demonstrated similar patterns to those found in earlier parachuting studies. ANOVAs demonstrated significant differences between sexes in self-report measures, but no significant differences in heart rate. In addition, examination of the relationships between heart rate and self-report showed a positive correlation for the women, while there was a negative correlation for the men.

Adolescent↗

PubMed1984

Seasonal changes of food and water consumption and urine production of the marmot, Marmota flaviventris.

In early spring, food and water consumption and the excretion and clearances of urine and solutes reached maximal rates. Water consumption exceeded food intake and urine production and plasma osmolality was lowest. Toward early and late summer, water intake decreased faster than food consumption and urine production. Urea excretion and clearances diminished with food consumption, while creatinine clearance decreased only slightly. Plasma osmolality increased. The data are consistent with rehydration soon after hibernation is completed, followed by a period of weight gain and dehydration in preparation for the next prolonged period of hibernation.

Animals↗

PubMed1984

Microsurgical anatomy of the recurrent artery of Heubner.

The recurrent arteries of Heubner were studied in 30 unfixed human brains (60 hemispheres) obtained from routine autopsies of individuals with a mean age of 34 years. The arteries were injected with tinted polyester resin via cannulation of the internal carotid arteries, and dissected under microscopic magnification. The recurrent artery of Heubner was absent in two hemispheres and double in seven hemispheres, either with a separate origin (in two) or from a common stem (in five). The artery of Heubner had a mean outer diameter of 0.8 +/- 0.04 mm (range 0.3 to 1.5 mm) and a mean length of 23.4 +/- 1.1 mm (range 12 to 38 mm). It originated from the A2 segment of the anterior cerebral artery (ACA) in 57% of the specimens, from the junction of the ACA and the anterior communicating artery in 35%, and from the A1 segment of the ACA in 8%. Three types of recurrent courses were observed. In the Type I or superior course, seen in 41 (63%) of the 65 arteries, the artery followed the superior wall of the A1 segment of the ACA. In the Type II or anterior course, the arteries found in 22 (34%) of specimens maintained a rostral position in relation to the A1 segment. In the Type III or posterior course, taken by two (3%) arteries, a posterior course of the vessel in the anterior perforated substance was found. The branching pattern was identified down to a range of 100 to 200 mu, and the average number of branches was 6.5 +/- 0.4 (range 3 to 12). Four groups of branches were observed. The olfactory group was represented in 91% by a single olfactory branch, with a mean outer diameter of 0.3 +/- 0.03 mm. On average, 1.9 +/- 0.27 frontal branches were found with a mean diameter of 0.13 +/- 0.01 mm. The branches penetrating the anterior perforated substance had a larger outer diameter (mean 0.4 +/- 0.03 mm) with an average number of 2.5 +/- 0.2 branches. The Sylvian fissure branches were more numerous (mean 3 +/- 0.3) with a mean outer diameter of 0.4 +/- 0.03 mm. The point of penetration of the main trunk was found to be constant at the level of the lateral perforated substance-medial Sylvian fissure in 85% of the cases. This report emphasizes the advantages of the intravascular casting resin injection technique in unfixed human brains over other conventional methods. It also describes the application of these anatomic data to the surgical strategy for the anterior circle of Willis, including the possible use of the recurrent artery of Heubner for microvascular reconstructive procedures.

Adult↗

PubMed1984

Microsurgical anatomy of the proximal segments of the middle cerebral artery.

The microvascular anatomy of the proximal segments (M1 and M2) of the middle cerebral artery (MCA) was studied in 70 unfixed brain hemispheres from 35 cadavers. The arteries were injected with a tinted polyester resin and dissected under magnification by microsurgical techniques. The authors studied the outer diameter (OD), length, site of origin, and pattern of branching of the main trunk, secondary trunks, and the initial insular portion of the cortical branches of the MCA. The degree of mobilization of the arteries lying over the insular cortex was also assessed. The main trunk of the MCA, which had an OD of 3 +/- 0.1 mm bilaterally and a length of 15 +/- 1.1 mm in the right hemisphere and 15.7 +/- 1.3 mm in the left hemisphere, could be divided into four groups: Group I: absence of a main division (that is, a single-trunk type of MCA) (in 6% of cases); Group II: bifurcation (64%); Group III: trifurcation (29%); and Group IV: quadrifurcation (1%). The secondary trunks resulting from the division of the main trunk of the MCA had a mean OD ranging from 1.4 to 2.3 mm and a mean length that varied from 12.1 to 14.9 mm. The mean OD of the cortical branches measured near their origin in the main and secondary trunks indicated that the angular artery was the largest vessel, with a mean OD of 1.5 mm on both sides of the brain. The temporopolar artery was the smallest, with a mean OD of 0.8 mm in the right hemisphere and 0.9 mm in the left hemisphere. The authors also describe the patterns of origin of the cortical vessels from the main trunk (early branches) and from the secondary trunks, as well as their branching pattern at the site of origin (single vessels and common stems). These anatomical data indicate that it is possible to perform microvascular reconstructive procedures, such as anastomosis, grafting, and reimplantation of branches in the insular area. The advantages of using unfixed specimens, intravascular injections, and magnification to reproduce in vivo conditions as closely as possible are also discussed.

Cerebral Arteries↗

PubMed1983

Rate-limiting step: a quantitative definition. Application to steady-state enzymic reactions.

The generality of the concept of a rate-limiting step in enzymic reactions recently has been questioned [Northrop, D. B. (1981) Biochemistry 20, 4056-4061] because, in simulated isotopic experiments, alterations of the step identified as rate limiting by current definitions do not consistently affect Vmax in the expected manner. In this paper a definition for a rate-limiting step is posed that eliminates such inconsistencies while the thrust of the original concept is retained. Thus, for any steady-state process involving a linear reaction sequence the rate-limiting step is taken as the "most sensitive" step, or the step which, if perturbed, causes the largest change in overall velocity, v. In both V and V/K enzymic systems the most sensitive step is identified by the relative magnitude of the sensitivity function, SFj, for the various forward steps. If forward steps are identified by kj, SFj is equal to delta(1/v)/[delta(1/kj)/(1/kj)], when the equilibrium constant for the step involving kj is maintained constant. The corresponding sensitivity index, SIj, is a normalized function of SFj (the normalizing factor is v) such that the sum of the values for SIj is equal to 1. In addition, there is an exact relationship between the sensitivity index for the isotopic step and the fraction of the intrinsic isotopic effect that is expressed in the overall rate of the reaction (when the intrinsic effect is taken as the fractional difference in reciprocal rate constant produced by the isotope). A procedure is described for approximating the sensitivity function for the various steps in a reaction sequence on the basis of the Gibbs energy profile for that reaction and thus identifying the most sensitive step. This approach also is used to consider the general question of whether a rate-limiting step should be specified for a multistep enzymic reaction. Identifying the rate-limiting step as the most sensitive step in a reaction sequence means that no aspect of the concept of minimal rate should be automatically considered as a property of a rate-limiting step.

Enzymes↗

PubMed1983

The complete amino acid sequence of rabbit muscle phosphoglucomutase.

The complete amino acid sequence of rabbit muscle phosphoglucomutase has been determined by isolating the 11 peptide fragments produced by the cyanogen bromide cleavage reaction and subjecting these to automated sequencing procedures. Products produced by treatment of some of these fragments with hydroxylamine, iodosobenzoic acid, mild acid, cyanogen bromide in formic and heptafluorobutyric acids, Staphylococcus aureus V8 protease, and trypsin (with or without blocking at lysine residues) were used to complete the sequence for each of the cyanogen bromide fragments. The cyanogen bromide fragments were ordered by isolating the four tryptic peptides produced by a limited tryptic digest of the native enzyme in the presence of its substrates and its bivalent metal ion activator, Mg2+, degrading these by means of trypsin, after blocking digestion at lysine residues, and isolating and identifying all fragments thus produced that contained 10 or more residues. The 561-residue sequence thus obtained is one of the longest that has been determined by chemical means. There is excellent agreement between this sequence and published compositions after appropriate normalization. The absorbance of the enzyme is about 7.0 at 278 nm for a 1% solution; this value is 9% lower than that previously used.

Amino Acid Sequence↗

PubMed1983

Factors that alter the plasma renin activity of the marmot.

The effects of low doses of norepinephrine (NE) and furosemide and a volume load (nonhibernators only) on plasma renin activity (PRA), mean arterial pressure (MAP), heart rate (HR), left renal (RBF) and right iliac (IBF) blood flow, cardiac index (CI), and total peripheral resistance (TPR) were determined in euthermic and hibernating marmots. In nonhibernating marmots NE produced an increase in CI and TPR and a decrease in RBF. In hibernators this dose of NE caused an increase in MAP, HR, and renal resistance, whereas it decreased PRA and did not alter iliac resistance. Furosemide infusions led to an increase in PRA in both groups and an increase in TPR in nonhibernators. The volume load in nonhibernators produced only a decrease in PRA. A comparison of control data from the two groups indicated that the renal and iliac beds contribute only a small portion to the increase in TPR that occurs during hibernation.

Animals↗

PubMed1982

Renal function of the pony and the horse.

Simultaneous renal clearances of inulin (CIN), p-aminohippurate (CPAH), and creatinine (CCR) were measured in hydrated mares (6 ponies and 2 horses). The CIN and CPAH were determined during steady-state infusion at 3 different infusion rates. A 6-fold change in plasma IN concentration did not produce alteration in CIN, nor was there a difference between the ponies and horses (P greater than 0.2). The overall average (mean +/- SEM) was 190.6 +/- 5.89 ml . min-1 . 100 kg of body weight-1. There was no difference noted between simultaneous CIN and CPAH. Clearance of PAH remained essentially constant during the change in plasma PAH from 0.33 mg/dl to 5.27 mg/dl. The extraction ratio of PAH for the nonanesthetized pony was 0.966. Effective renal plasma flow (CPAH) of the pony exceeded that of the horse.

Aminohippuric Acids↗

PubMed1981

The structure determination of rabbit phosphoglucomutase.

Tetragonal crystals of rabbit phosphoglucomutase have been grown from solutions containing ammonium sulphate, polyethylene glycol solution and enzyme. There are two molecules, each of relative molecular mass 64 000 per asymmetric unit. A rotation function suggests that these are related by a twofold axis. X-ray diffraction data for five heavy-atom derivatives and native crystals have been collected by using oscillation photography. A tentative and partial solution of the KAu(CN)2 sites has been obtained. The enzyme in the native crystals is phosphorylated, but the phosphate can be removed without harm to the crystals. Similarly the essential Mg2+ ion can be removed or replaced by Zn2+. The enzyme is active in the native crystals.

Animals↗

PubMed1981