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C R Meyer

Publications and source records attributed to C R Meyer.

At least 91 records · Page 5Linked to original sources
PubMed1992

Quantitative assessment of cartilage surface roughness in osteoarthritis using high frequency ultrasound.

Osteoarthritis (OA) is a common disease which affects nearly 50% of people over age 60. Histologic evaluation suggests that fibrillations approximately 20-150 microns are among the earliest changes in the articular cartilage. We propose a technique to quantify these surface fibrillatory changes in osteoarthritic articular cartilage by considering the angular distribution of the envelope-detected backscattered pressure field from an incident 30-MHz focused transducer. The angular distribution of the scattered acoustic field from an inosonifying source will directly relate to the distribution of surface fibrillatory changes. Data are presented for three different grades (400, 500 and 600 grit) of commercially available emory paper and three samples of osteoarthritic femoral head articular cartilage, which were visually assessed as having smooth, intermediate and rough surfaces, respectively. Our preliminary results indicate a probable monotonic relationship between articular cartilage roughening and the degree of broadening in the angle-dependent pressure amplitude. When applied to the emory paper, the technique indicates sensitivity to differences as small as approximately 5-10 microns in mean roughness. This procedure may provide an extremely sensitive and reproducible means of quantifying and following the cartilage changes observed in early osteoarthritis.

Cartilage, Articular↗
PubMed1991

Regulation of Phosphoenolpyruvate carboxylase from Crassula argentea: effect of incubation with ligands and dilution on oligomeric state, activity, and allosteric properties.

The relationship between the aggregation state and allosteric properties of purified phosphoenolpyruvate carboxylase from Crassula argentea was examined using both kinetic and physical techniques. Analysis by native polyacrylamide gel electrophoresis showed that dilution induced a dissociation of the active tetramer to a less active dimer. Kinetic assays showed that inhibition of phosphoenolpyruvate carboxylase by 5 mM malate measured at a saturating phosphoenolpyruvate concentration rose to nearly 80% with increasing preassay dilution while the activity in the absence of malate remained constant. Kinetic bursts were observed when enzyme-initiated assays were measured at a subsaturating phosphoenolpyruvate concentration. At saturating phosphoenolpyruvate concentrations, however, increasing lags developed in response to increasing the preassay dilution of the enzyme. Further, dynamic laser-light scattering measurements showed that preincubation of the dilute enzyme with phosphoenolpyruvate stabilized the tetramer while the presence of malate induced dimer formation. These observations confirm and extend earlier work with the extracted active malate insensitive night and less active, malate-sensitive day forms of the enzyme (Wu and Wedding [1985] Plant Physiol. 77, 667-675). Activity measured at subsaturating phosphoenolpyruvate concentrations dropped with increasing preassay dilution of enzyme, while activation by 3.2 mM glucose 6-phosphate, assayed at a low phosphoenolpyruvate concentration (0.044 mM), increased with dilution to nearly 400%. In this case activation results from a decrease in the control rate as the activity measured in the presence of glucose 6-phosphate was nearly constant, similar in effect to saturating phosphoenolpyruvate in the assay. Glucose 6-phosphate induced tetramer formation of the dilute enzyme as measured by light-scattering similar to the effects induced by PEP. In addition, when diluted (dimeric) PEPC was preincubated with PEP or glucose 6-phosphate the enzyme became less sensitive to malate inhibition, while the active-site directed ligand 2-phosphoglycolate had no effect on malate inhibition. These results indicate that both the substrate PEP and the activator glucose 6-phosphate stabilize the active tetramer via binding and interaction at an activator site separate from the active site.

Allosteric Regulation↗
PubMed1991

Pre- to postnatal reduction in ultrasound attenuation coefficient of the liver.

A recent study showed the ultrasound attenuation coefficient of fetal liver between 26 and 40 weeks of gestation to be 26% higher than after birth. To test the hypothesis that ultrasound attenuation is sensitive to fetal liver glycogen concentration, the livers of 24 fetuses were examined at 5 MHz just prior to and just after birth. The mean pre- to post-delivery reduction in attenuation coefficient was 0.08 dB cm-1 MHz-1 +/- 0.02 (SEM), or 17% of the post-delivery mean. This is consistent with the increase in attenuation measured by others in liver homogenate when glycogen was added. An increase in measurement accuracy, correlation with glycogen content, and, possibly, control for biological variability will be required to make predictions in individual cases, as opposed to these averages. A simple test of glycogen content would be of value scientifically and in prenatal and postnatal management.

Female↗
PubMed1991

Intravascular ultrasound imaging of vascular responsiveness in isolated perfused canine arteries.

We investigated the feasibility of using intravascular ultrasound imaging to analyze vascular physiology in various arterial beds. Canine superficial femoral, external iliac, and common carotid arteries were harvested and suspended and perfused in a bath of oxygenated, heated, physiologic salt solution. A 6-Fr, 20-MHz ultrasound imaging catheter was inserted into the lumen of the arteries and serial images were acquired after bolus injections of either serotonin or normal saline into the extravascular bathing medium. Serotonin resulted in a significant time- and dose-dependent decrease in cross-sectional area in muscular femoral arteries (P less than .001): -5.2% with 10(-8) M serotonin, -15% with 10(-7) M, and -28% with 10(-6) M. Histologically transitional iliac arteries demonstrated less marked changes, while elastic carotid arteries demonstrated no significant changes. Our results indicate that intravascular ultrasound may be used to quantify and differentiate responses to vasoconstrictive agents in different vascular beds.

Animals↗
PubMed1991

Fluorescence Study of Chemical Modification of Phosphoenolpyruvate Carboxylase from Crassula argentea.

The chemical modification of phosphoenolpyruvate carboxylase purified from Crassula argentea leaves was studied using the fluorescence of the extrinsic probe 8-anilino-1-naphalenesulfonate. The effects of ligands on kinetic parameters of phosphoenolpyruvate carboxylase activity, and its response to pH and metal cations, were associated with the binding of the ligands to the enzyme as measured by fluorescence. Binding of the ligands phosphoenolpyruvate, malate, and glucose-6-phosphate revealed by fluorescence measurements corresponds to competitive phenomena observed in kinetic studies. The fluorescence measurements also suggest the involvement of specific amino acids in the binding of a given ligand. Arginyl residues modified by 2,3-butanedione appear to be directly involved in the binding of phosphoenolpyruvate and malate to the active and the inhibition sites, respectively. A histidyl residue was involved in the binding of malate, accounting for the lack of inhibition by malate in kinetic studies of the enzyme treated with diethylpyrocarbonate. Although activity was lost, there was no decrease in the ability of the treated enzyme to bind phosphoenolpyruvate, suggesting that additional histidyl residues are essential for activity although not directly involved in the binding of phosphoenolpyruvate. The lysine reagent trinitrobenzenesulfonate caused a loss of activity and a reduction in malate inhibition and glucose-6-phosphate activation, but these modifications were not related to changes in the ability of the enzyme to bind any of the three ligands. This suggests that lysine residues were not directly involved in the binding of these ligands.

Journal Article↗
PubMed1990

The influence of pH on substrate form specificity of phosphoenolpyruvate carboxylase purified from Crassula argentea.

Purified phosphoenolpyruvate carboxylase from both the crassulacean acid metabolism plant Crassula argentea and the C4 plant Zea mays was shown by kinetic studies at saturating fixed-varying concentrations of free mg2+ to selectively use the metal-complexed form of phosphoenolpyruvate when assayed at pH 8.0. A similar response to added magnesium at high free phosphoenolpyruvate concentrations was obtained for both enzymes, consistent with the use of the complex as the substrate. Kinetic studies at pH 7.0 indicated that at this pH the total concentration of phosphoenolpyruvate (including both free and metal-complexed forms) could be used by the enzyme from C.argentea while the C4 enzyme still utilized the complex. The loss of specificity induced by the decrease in the pH of the assay medium was accompanied by a decrease in the Km of this enzyme for phosphoenolpyruvate whatever the form considered and an increase in Vmax/Km. In contrast, a similar decrease of pH led to an increased Km of the C4 enzyme for phosphoenolpyruvate and a decrease of Vmax/Km. For the enzyme from C. argentea (previously shown to contain an essential arginine at the active site), protection of activity by the different forms of substrate against inactivation by the specific arginyl reagent 2,3-butanedione changes markedly with pH. At pH 8.1, the metal complex is the better protector while at pH 7.0 free phosphoenolpyruvate gives the best protection consistent with the observed kinetic changes in substrate form utilization. The relationship between the enzyme affinity for substrate, substrate specificity, and the requirement for magnesium for substrate turnover is discussed.

Carboxy-Lyases↗
PubMed1990

The role of oligomerization in regulation of maize phosphoenolpyruvate carboxylase activity. Influence of Mg-PEP and malate on the oligomeric equilibrium of PEP carboxylase.

A purification procedure which yields a near homogenous preparation of phosphoenolpyruvate (PEP) carboxylase from the leaves of Zea mays is reported. The enzyme had a final specific activity of 33.3 micromoles per minute per milligram protein. Size exclusion high performance liquid chromatography and dynamic laser-light scattering spectroscopy showed that PEP carboxylase exists in an equilibrium of aggregates. Enzyme predominantly in the dimeric configuration is less active (when assayed at sub-optimal Mg-PEP concentrations, less than 0.4 millimolar) than when in its tetrameric arrangement. The difference in activity diminishes and disappears as the concentration of the substrate Mg-PEP increases. The substrate drives the equilibrium toward the tetramer, while malate, an inhibitor of PEP carboxylase, shifts the equilibrium toward the dimer. It thus appears that the quaternary structure (oligomeric state) of maize PEP carboxylase can be regulated by the naturally occurring effector molecules Mg-PEP and malate which in turn can control the enzyme's activity.

Chromatography, High Pressure Liquid↗
PubMed1990

Regulation of the aggregation state of maize phosphoenolpyruvate carboxylase: evidence from dynamic light-scattering measurements.

The molecular weights of different aggregational states of phosphoenolpyruvate carboxylase purified from the leaves of Zea mays have been determined by measurement of the molecular diameter using a Malvern dynamic light scattering spectrometer. Using these data to identify the monomer, dimer, tetramer, and larger aggregate(s) the effect of pH and various ligands on the aggregational equilibria of this enzyme have been determined. At neutral pH the enzyme favored the tetrameric form. At both low and high pH the tetramer dissociated, followed by aggregation to a "large" inactive form. The order of dissociation at least at low pH appeared to be two-step: from tetramer to dimers followed by dimer to monomers. The monomers then aggregate to a large aggregate, which is inactive. The presence of EDTA at pH 8 protected the enzyme against both inactivation and large aggregate formation. Dilution of the enzyme at pH 7 at room temperature results in driving the equilibrium from tetramer to dimer. The presence of malate with EDTA stabilizes the dimer as the predominant form at low protein concentrations. The presence of the substrate phosphoenolpyruvate alone and with magnesium and bicarbonate induced formation of the tetramer, and decreased the dissociation constant (Kd) of the tetrameric form. The inhibitor malate, however, induced dissociation of the tetramer as evidenced by an increase in the Kd of the tetramer.

Carboxy-Lyases↗
PubMed1990

Ultrasound attenuation coefficient in the fetal liver as a function of gestational age.

An apparent increase in the ultrasound attenuation coefficient per unit frequency, alpha f, of fetal liver as a function of gestational age has been observed. Measurements were made in utero with a 25 megasample/sec RF digitizer and a real time ultrasound system with a 5 MHz scan head. A precise measurement of alpha f was employed in which the intercept was tied to 0 at a frequency of 0. In 178 examinations of normal pregnancies, the linear regression of the alpha f increased 26% between 26 and 40 weeks gestation. This statistically significant increase (p less than 0.0001) is consistent with several observations, those of Parker et al. of increased attenuation in liver when glycogen is added, the increasing glycogen storage in the liver before birth, and our own pre- and postnatal measurements reported elsewhere. A noninvasive assay for glycogen content would have important applications in medicine and biomedical science. However, an increase in measurement accuracy and precise correlation with glycogen content will be required to make meaningful predictions in individual cases, as opposed to the present statistical trends.

Embryonic and Fetal Development↗
PubMed1990

Inhibition of phosphoenolpyruvate carboxylase by malate.

Malate has been noted to be a ;mixed' inhibitor of phosphoenolpyruvate (PEP) carboxylase. The competitive portion of this inhibition appears to be fairly constant regardless of the condition of the enzyme being measured, but the noncompetitive (V-type) inhibition is subject to variation depending on the source of the enzyme, its storage condition, the presence or absence of various ligands, and differences in pH. In the case of the maize (Zea mays L.) phosphoenolpyruvate carboxylase (PEPC), the V-type inhibition by malate is much less pronounced at pH 8 than at pH 7. Examination of the response of the maize PEPC to PEP concentration reveals a pronounced cooperativity at pH 8 which is not present at pH 7, and which results in the disappearance of the V-type inhibition at pH 8. The ability of high concentrations of PEP to convert PEPC from a form readily inhibited by malate to one resistant to malate inhibition has been previously demonstrated and we attribute the cooperativity shown at pH 8 to this response to high levels of PEP. Support for this proposal is provided by studies of the enzyme at pH 7 and pH 8 run in 20% glycerol. In this case there was no V-type inhibition of PEPC at either pH. Treatment with 20% glycerol has been shown to result in the aggregation of maize PEPC.

Journal Article↗
PubMed1989

A kinetic study of the effects of phosphate and organic phosphates on the activity of phosphoenolpyruvate carboxylase from Crassula argentea.

The effects of phosphate and several phosphate-containing compounds on the activity of purified phosphoenolpyruvate carboxylase (PEPC) from the crassulacean acid metabolism plant, Crassula argentea, were investigated. When assayed at subsaturating phosphoenolpyruvate (PEP) concentrations, low concentrations of most of the compounds tested were found to stimulate PEPC activity. This activation, variable in extent, was found in all cases to be competitive with glucose 6-phosphate (Glc-6-P) stimulation, suggesting that these effectors bind to the Glc-6-P site. At higher concentrations, depending upon the effector molecule studied, deactivation, inhibition, or no response was observed. More detailed studies were performed with Glc-6-P, AMP, phosphoglycolate, and phosphate. AMP had previously been shown to be a specific ligand for the Glc-6-P site. The main effect of Glc-6-P and AMP on the kinetic parameters was to decrease the apparent Km and increase Vmax/Km. AMP also caused a decrease in the Vmax of the reaction. In contrast, phosphoglycolate acted essentially as a competitive inhibitor increasing the apparent Km for PEP and decreasing Vmax/Km. Inorganic phosphate had a biphasic effect on the kinetic parameters, resulting in a transient decrease in Km followed by an increase of the apparent Km for PEP with increasing concentration of phosphate. The Vmax also was decreased with increasing phosphate concentrations. Further, the enzyme appeared to respond to the complex of phosphate with magnesium. In the presence of a saturating concentration of AMP, no activation but rather inhibition was observed with increasing phosphate concentration. This is consistent with the binding of phosphate to two separate sites--the Glc-6-P activation site and an inhibitory site, a phenomenon that may be occurring with other phosphate containing compounds. High concentrations of phosphate with magnesium were found to protect enzyme activity when PEPC, previously shown to contain an essential arginine at the active site, was incubated with the specific arginyl reagent 2,3-butanedione, consistent with the binding of phosphate at the active site. Data were successfully fitted to a rapid equilibrium model allowing for binding of the phosphate-magnesium complex to both the activation site and the active site which accounts for the activation/deactivation observed at low substrate concentrations. Effects on the Vmax of the reaction are also addressed. Factors controlling the differential affinity of various effectors to the active site or activation site appear to include charge distribution, size, and other steric factors.

Adenosine Monophosphate↗
PubMed1989

Assessment of ultrasonic computed tomography in symptomatic breast patients by discriminant analysis.

From 95 subjects imaged with both speed of sound and attenuation ultrasonic computed tomography (UCT), quantitative analyses are presented on 40 cases where unequivocal correlating clinical diagnoses are available. Using four attenuation and speed of sound parameters from different regions of interest in the breast, a linear discriminator detects cancer with approximately 90% sensitivity and specificity. Increased confidence in the predictive power of this small study is given by a modern test of predictive power (jackknifing) and by the fact that diagnostic discrimination remains as high as 85% when only two parameters are employed--attenuation and speed of sound in the lesion minus those values in the remaining central mammary tissues. Speed of sound images appear particularly useful in older, fatty breasts where pulse echo ultrasound is particularly lacking. While UCT in the form studied here is not likely to receive wide clinical acceptance in the near future, a combined UCT/pulse echo system might find wide clinical utility if it can be sufficiently convenient and inexpensive.

Adenofibroma↗
PubMed1989

Liver-tumor boundary detection: human observer vs computer edge detection.

As a preliminary step in computing tumor volume, we developed a computer edge detection program to define the liver-tumor interface in computed tomography (CT) images. Computer program performance was tested using CT images from a lucite liver/tumor phantom; from normal livers containing computer-generated pseudotumors of known size, object contrast, and liver-tumor edge gradients; and from 12 abdominal livers containing 19 focal tumors, eight with well-defined and 11 with ill-defined borders. Calculated sizes of the tumor phantom and pseudo-tumors were compared with measured volumes and predetermined cross-sectional areas, respectively. In the absence of a truth standard for the size of the focal hepatic tumors, computer-calculated cross-sectional areas of the tumors were compared with the measurements made by an experienced interpreter of CT images using the trackball cursor at the CT console. The console measurements were made five times on separate days during a one-week period. The variability in the measured areas of these tumors averaged 7.1% for the well-defined tumors and 14.0% for the poorly defined tumors (P = 0.05). The edge-linking algorithm systematically overestimated the volumes of individual slices of the hemispherical tumors in the lucite phantom. Nevertheless, because of algorithm failure in the slices containing the poles of the hemispheres, errors in total tumor volumes were -2.1% for the 5.1 cm radius tumor, +1.2% for the 2.7 cm radius tumor, and +15% for the 1.8 cm radius tumor. The edge-linking algorithm was reasonably successful in calculating areas of pseudotumors with object contrast of 3.0% or greater and steep edge gradients.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗
PubMed1989

Activation of higher plant phosphoenolpyruvate carboxylases by glucose-6-phosphate.

Studies of the response of phosphoenolpyruvate carboxylase from C(3) (wheat [Triticum aestivum L.]), C(4) (maize [Zea mays L.]), and Crassulacean acid metabolism (CAM) (Crassula) leaves to the activator glucose-6-phosphate as a function of pH showed that the binding of the activator and the response path to activation were essentially identical for all three enzymes. The level of affinity for the activator differed, with the CAM enzyme having the highest affinity and the maize enzyme the lowest. The observed pK values suggest that histidine and cysteine groups may be involved in activation by glucose-6-phosphate. The presence of glucose-6-phosphate protected the enzyme against inactivation of the activation response by p-chloromercuribenzoate. The maximal activation response to glucose-6-phosphate showed differences among the three enzymes including different pH optima and different pH profiles. Here the maize leaf enzyme showed a potential response about twice as great as that of the C(3) and CAM enzymes.

Journal Article↗
PubMed1989

A crisis in spirit.

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Attitude of Health Personnel↗
PubMed1988

Identification of substrate and effector binding sites of phosphoenolpyruvate carboxylase from Crassula argentea. A possible role of phosphoenolpyruvate as substrate and activator.

Phosphoenolpyruvate carboxylase (EC 4.1.1.31) purified from leaves of the crassulacean acid metabolism plant (Crassula argentea) was chemically modified by the specific arginyl reagent 2,3-butanedione. Modification resulted in enzyme inactivation which followed pseudo first-order kinetics. Participation of arginyl residues involved in the binding of or response to both phosphoenolpyruvate and malate, respectively, was established. Inactivation and protection studies suggest the presence of three sites involved in the binding of the substrate, phosphoenolpyruvate, the activator, glucose 6-phosphate, and the inhibitor, malate. Studies using both fluorescence measurements of binding and steady-state kinetic methods indicate that phosphoenolpyruvate can bind both to the active site and to the activator site. Evidence for stimulation of the activity of phosphoenolpyruvate carboxylase upon the binding of substrate to the activation site was provided by kinetic studies using AMP, previously shown to be a specific ligand for the activation site.

Carboxy-Lyases↗