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W J Ray

Publications and source records attributed to W J Ray.

At least 73 records · Page 4Linked to original sources

Microanatomy of the anterior cerebral artery.

The microanatomic features of the anterior cerebral artery were studied in 30 unfixed human brains which were injected with tinted polyester resin via cannulation of the internal carotid arteries under microscopic dissection. The outer diameter, length, and number of perforating branches were measured for each of the following vessels: anterior cerebral artery (proximal A1 segment, distal A2 segment), anterior communicating artery, and recurrent artery of Heubner. The perforating branches of the proximal segment of the anterior cerebral artery penetrated the brain at the anterior perforated substance, lateral chiasm, and optic tracts. The perforating branches of the anterior communicating artery penetrated the brain at the lamina terminalis, anterior perforated substance, and medial chiasm. The first 5 mm of the distal anterior cerebral artery (A2) had perforating branches penetrating the brain at the gyrus rectus and olfactory sulcus. The recurrent artery of Heubner originated from the A2 segment of the anterior cerebral artery in 57% of the cases, from the anterior cerebral artery-anterior communicating artery junction in 35%, and from the A1 segment in 8%. The depth of the interhemispheric fissure at the genu was 36.0 +/- 0.5 mm and at the midbody of the corpus callosum, 35.0 +/- 0.5 mm. Extension of the dissection to approach the anterior communicating artery from the genu of the corpus callosum using the anterior interhemispheric route was an additional 31.7 +/- 0.7 mm. The callosal arterial supply from the anterior cerebral artery showed short callosal branches in all brain specimens and long callosal vessels in 10% of the specimens.

Cerebral Arteries↗

Active-site serine phosphate and histidine residues of phosphoglucomutase: pH titration studies monitored by 1H and 31P NMR spectroscopy.

1H and 31P NMR pH titrations were conducted to monitor changes in the environment and protonation state of the histidine residues and phosphoserine group of rabbit muscle phosphoglucomutase on binding of metal ions at the activating site and of substrate (glucose phosphate) at the catalytic site. Imidazole C epsilon-H signals from 8 of the 10 histidines present in the free enzyme were observed in 1H NMR spectra obtained by a spin-echo pulse sequence at 470 MHz; their pH (uncorrected pH meter reading of a 2H2O solution measured with a glass electrode standardized with H2O buffer) titration properties (in 99% 2H2O) were determined. Three of these histidine residues, which have pKa values ranging from 6.5 to 7.9, exhibited an atypical pH-dependent perturbation of their chemical shifts with a pHmid of 5.8 and a Hill coefficient of about 2. Since none of the observed histidines has a pKa near 5.8, it appears that these three histidines interact with a cluster consisting of two or more groups which become protonated cooperatively at this pH. Binding of Cd2+ at the activating site of the enzyme abolishes the pH-dependent transition of these histidines; hence, the putative anion cluster may constitute the metal ion binding site, or part of it. Two separate 31P NMR peaks from phosphoserine-116 of the phosphoenzyme were observed between pH 6 and 9. Apparently, the metal-free enzyme exists as a pH-dependent mixture of conformers that provide two different environments, I and II, for the enzymic phosphate group; the transition of the phosphate group between these two environments is slow on the NMR time scale.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

EEG alpha activity reflects attentional demands, and beta activity reflects emotional and cognitive processes.

Two experiments were designed to examine the effects of attentional demands on the electroencephalogram during cognitive and emotional tasks. We found an interaction of task with hemisphere as well as more overall parietal alpha for tasks not requiring attention to the environment, such as mental arithmetic, than for those requiring such attention. Differential hemispheric activation for beta was found most strongly in the temporal areas for emotionally positive or negative tasks and in the parietal areas for cognitive tasks.

Alpha Rhythm↗

Multinuclear magnetic resonance studies of metal ion binding sites of phosphoglucomutase.

Metal binding at the activating site of rabbit muscle phosphoglucomutase has been studied by 31P, 7Li, and 113Cd NMR spectroscopy. A 7Li NMR signal of the binary Li+ complex of the phosphoenzyme was not observed probably because of rapid transverse relaxation of the bound ion due to chemical exchange with free Li+. The phosphoenzyme-Li+-glucose 6-phosphate ternary complex is more stable, kinetically, and yields a well-resolved peak from bound Li+ at -0.24 ppm from LiCl with a line width of 5 Hz and a T1 relaxation time of 0.51 +/- 0.07 s at 78 MHz. When glucose 1-phosphate was bound, instead, the chemical shift of bound 7Li+ was -0.13 ppm; and in the Li+ complex of the dephosphoenzyme and glucose bisphosphate a partially broadened 7Li+ peak appeared at -0.08 ppm. Thus, the bound metal ion has a somewhat different environment in each of these three ternary complexes. The 113Cd NMR signal of the binary Cd2+ complex of the phosphoenzyme appears at 22 ppm relative to Cd(ClO4)2 with a line width of 20 Hz at 44.4 MHz. Binding of substrate and formation of the Cd2+ complex of the dephosphoenzyme and glucose bisphosphate broaden the 113Cd NMR signal to 70 Hz and shift it to 75 ppm. The 53 ppm downfield shift upon the addition of substrate along with 1H NMR data suggests that one oxygen ligand to Cd2+ in the binary complex is replaced by a nitrogen ligand at some intermediate point in the enzymic reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

A simple procedure for removing contaminating aldehydes and peroxides from aqueous solutions of polyethylene glycols and of nonionic detergents that are based on the polyoxyethylene linkage.

Commercial samples of polyethylene glycol as well as detergents incorporating the polyoxyethylene linkage may contain substantial concentrations of peroxides and aldehydes, as well as aldehyde precursors. Facile procedures for essentially eliminating either the first or all three of these contaminants from concentrated aqueous stock solutions are described. Under some conditions, the purified solutions can be stored at room temperature under aerobic conditions for time periods of weeks without significant decomposition. Possible antioxidants that might be used under other conditions to stabilize such solutions during long-term aerobic studies are considered.

Aldehydes↗

EEG correlates of emotional tasks related to attentional demands.

This research brings together two separate areas: that of EEG processes associated with positive and negatively valenced emotional material; and that of traditional psychophysiological research related to the "intake" and "rejection" of environmental stimuli. Forty males on each of two days were presented with tasks reflecting both attentional demands and affectual processing. Heart rate and bilateral EEG measures from frontal, parietal and temporal sites were recorded. Using a FFT (fast Fourier transform) electrocortical activity in the 2-7 Hz, 8-15 Hz, and 16-24 Hz was determined and analyzed. The results suggest emotional valence (i.e. positive and negative) and attentional demands (i.e. intake vs rejection) are differentially represented in terms of EEG functioning. An interaction of attentional demand with hemisphere was found for EEG alpha activity in the temporal and parietal areas. For emotional valence there was a significant main effect for EEG beta activity in both the temporal and parietal areas. Differential hemispheric activity was found using a factor analytic technique (PARAFAC) with positively valenced tasks being associated with right temporal beta. Heart rate changes for the attentional dimension were consistent with previous research.

Adolescent↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗