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Biomedical subjects

D C Wood

Publications and source records attributed to D C Wood.

At least 37 records · Page 2Linked to original sources
PubMed1991

Diagnosis, transport, and outcome in fetuses with left ventricular outflow tract obstruction.

Between March 1986 and April 1990, 22 consecutive fetuses (at gestational ages of 21 to 38 weeks) with a suspected diagnosis of critical (ductus-dependent) left ventricular outflow tract obstruction on fetal echocardiogram were referred to our center for delivery and surgical treatment. Diagnoses were hypoplastic left heart syndrome (n = 16), valvular aortic stenosis (n = 2), common atrioventricular canal with subaortic stenosis (n = 3), and single ventricle with subaortic stenosis (n = 1). Postnatal echocardiography revealed that fetal echocardiography was correct in predicting left ventricular outflow tract obstruction to be critical in all but one patient, for a positive predictive value of 96%. Of the 21 patients with true, critical left ventricular outflow tract obstruction, 17 patients underwent cardiac surgery as neonates (birth to 6 days of age, median 2 days); 13 (or 77%) survived and were discharged from the hospital. In addition, one patient underwent successful balloon aortic valvotomy for critical valvular aortic stenosis but later died of sepsis. Lethal chromosomal and congenital abnormalities should be sought and are contraindications for this approach. In utero transport of fetuses with suspected critical left ventricular outflow tract obstruction to a neonatal cardiac surgical center can result in improved neonatal condition and may improve overall survival.

Echocardiography, Doppler↗
PubMed1989

Purification and characterization of pituitary bovine somatotropin.

Bovine somatotropin (bST) has been isolated from pituitary glands and compared in a variety of chemical analyses and bioassays with somatotropin derived from recombinant Escherichia coli. Comparison of pituitary extracts and purified bST by Western blot analysis of two-dimensional gels suggested that the immunoreactive somatotropin species present in the extract were also present in the purified material, with no significant losses or degradation as a result of the purification method. NH2-terminal sequence analysis indicated the presence of equal quantities of Ala-Phe-Pro-Ala-Met-Ser-Leu-Ser- and Phe-Pro-Ala-Met-Ser-Leu-Ser- sequences. The Met-Ser-Leu-Ser-NH2-terminal sequence, a degradation product observed in NIH standard lots, was not detected. Assay of bioactivity in a bovine liver receptor-binding assay and in a female rat growth assay showed pituitary bST and recombinant methionyl-bovine somatotropin to be equipotent. Tryptic maps and sequence analysis of pituitary-derived somatotropin suggest the presence of isoaspartate derivatization at Asp128.

Amino Acid Sequence↗
PubMed1989

Localization of mechanoreceptors in the protozoan, Stentor coeruleus.

Mechanoreceptor channels were localized by using the ligands, tubocurarine (TC), decamethonium (Deca), and gallamine (Gall), which have been shown to bind specifically to these channels. The binding of radioactively labeled TC (TC*) was found to be directly proportional to the cell surface area suggesting that the channels are uniformly distributed over the cell surface. Intracellular TC and Gall injections did not depress mechanical stimulus sensitivity though these drugs did depress sensitivity when applied extracellularly at the same concentrations; therefore, the ligand binding sites are on or near the external surface of the cell. Autoradiographs revealed that radioactively labeled Deca (Deca*) bound to the pigmented stripes but not to the ciliary stripes or membranellar band. Further, Stentor induced to shed their membranellar band through exposure to 8% urea were more sensitive to mechanical stimuli than were controls; therefore, the membranellar cilia do not appear to contain mechanoreceptor channels. Collectively, these data indicate that the mechanoreceptor channels are located in the somatic surface covering the pigmented stripes. The density of mechanoreceptor channels in the plasma membrane covering the somatic surface is tentatively estimated to be between 5500 and 14,500 microns-2 based on the density of TC* binding, the apparent number of TC molecules binding per mechanoreceptor channel, and data suggesting that only one fifth to one fourth of the bound TC* is bound to structures in the plasma membrane.

Animals↗
PubMed1988

Habituation in Stentor: a response-dependent process.

The contractile protozoan Stentor coeruleus habituates during repetition of the mechanical stimulus used to elicit the initial contractions. This decrement in response probability was found to be highly correlated with a reduction in receptor potential amplitude, while the amplitude of the action potentials that triggered the contractions did not change. When mechanical stimulation elicited receptor potentials and not action potentials, the receptor potential did not habituate significantly. Conversely, action potentials repetitively elicited by current pulses habituated animals to mechanical stimuli. Similarly, voltage steps used to simulate action potentials produced pronounced decrements in receptor currents recorded from voltage-clamped cells, while mechanical stimulation produced only small decrements. Thus, habituation depends primarily on action potential production, while mechanical stimulation itself makes a much smaller, but significant, contribution. The temporal relation between mechanical stimuli and action potentials, when both occur, is inconsequential in determining the rate and degree of habituation produced.

Animals↗
PubMed1988

Habituation in Stentor: produced by mechanoreceptor channel modification.

Habituation in Stentor was shown in the previous paper (Wood, 1988) to result from a progressive diminution in mechanoreceptor potential amplitude during the course of repetitive mechanical stimulation. The source of this diminution was analyzed in the present studies by eliciting mechanoreceptor currents from control and habituated cells under voltage clamp control. The I-V plots obtained have a voltage-dependent region between -60 and -20 mV, and a linear voltage-independent region between -20 and 20 mV, with a reversal potential near 20 mV. The slope of the linear region did not change as a result of habituation indicating that the maximum mechanoreceptor current, and hence the total number of mechanoreceptor channels, was the same in control and habituated cells. The mechanoreceptor current reversal potential was changed by only a few millivolts, from 21.6 to 17.2 mV, leaving the driving force term for the mechanoreceptor current at resting potential relatively unchanged. On the other hand, the voltage-dependent region of the I-V plot became significantly steeper after habituation (9.6 mV/e-fold change) compared to its value before habituation (12.3 mV/e-fold change). This shift in voltage dependence is sufficient to explain the diminution in receptor potential amplitude seen during normal recordings at resting potential. The importance of the voltage-dependent characteristic of the mechanoreceptor channels to the habituation process was corroborated by the observation that the amplitude of receptor potentials elicited from cells depolarized by current pulses habituated to only a very limited degree while receptor potentials elicited at resting potential underwent marked habituation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

An improved method for measurement of common tricyclic antidepressant drugs and their metabolites using normal phase HPLC.

A sensitive HPLC method that allows the simultaneous determination of imipramine, amitriptyline and their desmethyl metabolites, using a 10 cm straight silica cartridge column with a mobile phase consisting of 0.004% perchloric acid in methanol is presented. Detection of the eluent is at 210 nm. The drugs are back-extracted from serum, giving both a good recovery and sensitivity of at least 5 micrograms/L. This method has proved to be extremely robust and reliable, with coefficients of variation around 3% being reported.

Antidepressive Agents, Tricyclic↗
PubMed1985

The mechanism of tubocurarine action on mechanoreceptor channels in the protozoan Stentor coeruleus.

(+)-Tubocurarine (TC) decreases the probability that the protozoan, Stentor coeruleus Ehrenberg, will contract in response to mechanical stimulation, because it selectively depresses mechanoreceptor currents. Resting membrane properties and action potentials are not significantly altered by the drug. Stentor incubated in media containing radioactively labelled TC (TC*) retain TC* after extensive washing despite a rather high apparent KD (19.7 mumol l-1). The incubation curve for TC* binding exhibits an initial exponential rise followed by a linear increase. Wash-out of bound TC* and elimination of the exponential component of the incubation curve is observed if the TC* incubation is followed by a 5-s exposure to 8% urea; therefore, the exponential component represents a reversible binding process. TC* binding in the exponential component is highly correlated (r less than -0.96) with the depression in receptor current and response probability when incubation time, drug concentration and drug (gallamine, TC, decamethonium and succinylcholine) are varied. These correlations suggest that the exponential binding is to functional mechanoreceptors. Mechanoreceptor currents are decreased by hyperpolarization and increased by depolarization, indicating that the mechanoreceptor channel is voltage-dependent. At hyperpolarized potentials the channels are in a form (the U form) which cannot be opened by mechanical stimulation; at depolarized potentials they are in a form (the R form) which can be opened. TC appears to bind to the U form with higher affinity than to the R form, since depolarization reduces the amount of bound TC* and relieves the depression of mechanoreceptor current produced by TC.

Action Potentials↗
PubMed1984

Effect of supercoiling on the abortive initiation kinetics of the RNA-I promoter of ColE1 plasmid DNA.

An analysis of the effect of DNA supercoiling on transcription of the RNA-I promoter of ColE1 has been undertaken using the abortive initiation assay developed by McClure (McClure, W. R., Cech, C. L., and Johnston, D. E. (1978) J. Biol. Chem. 253, 8941-8948). A tau plot was constructed, enabling the equilibrium constant for closed complex formation (KI) and the isomerization rate constant (k2) to be determined. The effect of supercoiling on RNA-I is an apparent 100-fold increase in the isomerization rate, k2, with little or no effect on KI. This is the first reported abortive initiation analysis of RNA-I transcription which serves in the regulation of plasmid replication.

DNA, Bacterial↗
PubMed1981

The N-ethylmaleimide-sensitive cysteine residue in the pH-dependent subunit interactions of malate dehydrogenase.

The specific chemical modification by N-ethylmaleimide of a cysteine residue at pH 5.0 in porcine heart mitochondrial malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37) has been shown to result in an enzymatically inactive, monomeric product, which does not reassociate at pH 7.5 to yield the native dimer. In this report, an investigation of proton release and uptake upon NADH binding to the native enzyme and to the N-ethylmaleimide-modified enzyme has implicated the above cysteine residue as being directly linked to the pH-dependent subunit dissociation of mitochondrial malate dehydrogenase. The results are consistent with the view that the modified cysteine residue is not located at the subunit interaction site, although it is probably near this site. A recent study from this laboratory has demonstrated that the monomeric enzyme obtained at pH 5.0 exists in a conformation which is enzymatically inactive and which has an enhanced intrinsic protein fluorescence. Interpretation of protein fluorescence data has suggested that the N-ethylmaleimide modification results in inactivation of the enzyme by preventing the pH-induced conformational change to the active dimer. However, NADH is able to induce reassociation of the N-ethylmaleimide-modified enzyme at pH 7.5 but not at pH 5.0. This reassociation at pH 7.5 is accompanied by a significant regain of enzymatic activity, indicating that NADH binding is able to partially overcome the negative effect of the cysteine modification on the pH-dependent subunit reassociation of mitochondrial malate dehydrogenase.

Animals↗
PubMed1981

Subunit interactions in mitochondrial malate dehydrogenase. Kinetics and mechanism of reassociation.

The pH-dependent dissociation of porcine heart mitochondrial malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37) has been more extensively characterized. The native, dimeric form of the enzyme (Mr = 70,000) which exists at pH 7.5 has previously been shown to dissociate into its constituent subunits (Mr = 35,000) at pH 5.0 (Bleile, D. M., Schulz, R. A., Gregory, E. M., and Harrison, J. H. (1977) J. Biol. Chem. 252, 755-758). The dissociation is accompanied by a concomitant decrease in enzymatic specific activity and an increase in intrinsic protein fluorescence. By using the characteristics of specific activity and intrinsic protein fluorescence as probes of dimerization, the kinetics of subunit reassociation was investigated. In order to facilitate reassociation, a pH jump method was utilized in which enzyme at pH 5.0 was diluted into a large excess of pH 7.5 buffer. The regain of enzymatic specific activity and the decrease in protein fluorescence were observed to follow first order kinetics. The rate constant in both cases was dependent upon the protein concentration, and in all cases, full recovery of either enzymatic activity or native protein fluorescence was obtained. The Arrhenius activation energy for the reassociation of the subunits was found to be approximately 20 kcal/mol, an observation which is consistent with a refolding process whose rate-limiting step may be the cis/trans-isomerization about one or more proline imino bonds. A model for subunit reassociation which is consistent with the kinetic data is proposed.

Animals↗
PubMed1981

The immobilization of mitochondrial malate dehydrogenase on Sepharose beads and the demonstration of catalytically active subunits.

Porcine heart mitochondrial malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37) has been immobilized by covalent attachment to CNBr-activated Sepharose 4B-Cl gel. The gel was activated with low levels of CNBr to produce a low density of linkage sites and, hence, to facilitate linkage of the enzyme through a single subunit. Matrix-bound mitochondrial malate dehydrogenase was found to possess 50-65% of the native mitochondrial malate dehydrogenase specific activity when assayed in the NAD+ leads to NADH direction but only 5-15% of the native enzyme specific activity when assayed in the NADH leads to NAD+ direction. MB-dimeric mitochondrial malate dehydrogenase was dissociated to MB-monomer by exposure to pH 5.0 buffer. The MB-monomer was found to be catalytically active, possessing only a slightly decreased specific activity when compared to MB-dimer. The reconstitution of Mb-monomer to MB-dimer was accomplished by adding dissociated mitochondrial malate dehydrogenase, which exists at pH 5.0, to MB-monomer and adjusting to pH 7.5. The kinetic parameters, pH activity profile, and stability toward heat denaturation for MB-mitochondrial malate dehydrogenase (monomer and dimer) were determined and compared to native mitochondrial malate dehydrogenase. MB-mitochondrial malate dehydrogenase exhibited enhanced stability and similar pH activity profiles when compared to native mitochondrial malate dehydrogenase. Immobilization of mitochondrial malate dehydrogenase altered the enzyme's kinetic parameters in such a manner as to increase the values of Km for the substrates and decrease the values of Vmax.

Animals↗
PubMed1981

Radiation-induced parotid cancer.

A retrospective analysis of 72 cases of primary malignant tumors of the parotid gland treated at the University of Illinois Hospital, Chicago, from 1950 through 1978 revealed that six of these had developed from two to 24 years after irradiation of the head or neck for various benign and malignant neoplastic conditions. At the time of irradiation, ages ranged from 7 to 73 years; the sex distribution was equal. From our findings and those in 26 cases reported by various other authors, the following criteria are proposed for the designation of a parotid tumor as being radiation induced: (1) well-documented radiation exposure; (20 port of irradiation must incorporate the gland in which the cancer subsequently arises; (3) exposure to a minimum of 300 rads; and (4) minimum latent period of two years. In view of the widespread use in the past of head and neck irradiation of benign neoplastic disease, the surgeon should be aware of this possible link with parotid gland tumor.

Adenocarcinoma↗