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Biomedical subjects

D E Brown

Publications and source records attributed to D E Brown.

At least 55 records · Page 3Linked to original sources

Resonance Raman spectroscopy of methylamine dehydrogenase from bacterium W3A1.

Resonance Raman spectroscopy has been used to probe the structure of the covalently bound quinone cofactor in methylamine dehydrogenase from the bacterium W3A1. Spectra were obtained on the phenylhydrazine and 2-pyridylhydrazine derivatives of the native enzyme, on the quinone-containing subunit labeled with phenylhydrazine, and on an active-site peptide also labeled with phenylhydrazine. Comparisons of these spectra to the corresponding spectra of copper-containing amine oxidase derivatives indicate that the quinones in these two classes of quinoproteins are not identical. The resonance Raman spectra of the native enzyme and small subunit have also been measured. 16O/18O exchange permitted the carbonyl modes of the quinone to be identified in the resonance Raman spectrum of oxidized methylamine dehydrogenase: a band at 1614 cm-1, together with a shoulder at 1630 cm-1, are assigned as modes containing substantial C = O stretching character. D2O/H2O exchange has pronounced effects on the resonance Raman spectrum of the oxidized enzyme, suggesting that the quinone may have numerous hydrogen bonds to the protein or that it is unusually sensitive to the local environment. Resonance Raman spectra of the isolated small subunit, and its phenylhydrazine derivative, are considerably different from the corresponding spectra of the intact protein. An attractive explanation for these observations is that the quinone cofactor in methylamine dehydrogenase from W3A1 is located at the interface between the large and small subunits, as found for the enzyme from Thiobacillus versutus [Vellieux, F. M. D., Huitema, F., Groendijk, H., Kalk, K. H., Frank, J. Jzn., Jongejan, J. A., & Duine, J. A. (1989) EMBO J. 8, 2171-2178].(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteria↗

An optically scanned EMS reporting form and analysis system for statewide use: development and five years' experience.

Analysis of emergency medical services (EMS) systems data is crucial to planning, education, research, and quality assurance programs. Currently, comparative analysis of EMS data between regions or states is virtually impossible due to wide variations in data collection and analysis methods. To devise a practical and uniform EMS reporting system, we referenced the minimum data set (MDS) established by the federal government in 1974 and surveyed 22 states known to be using uniform reporting systems. In developing our final data set, elements were added based on inclusion in the MDS, national survey results, a review of current EMS literature, and consensus of local EMS providers. This set of 48 elements then was incorporated into a reporting form using narrative and optically scanned formats, allowing automated data collection for computer analysis. After a pilot study, the system was improved to allow high-speed ink reading and large volume data storage and analysis using a microcomputer. This system has subsequently been adopted by seven states. The combined data base exceeds 250,000 cases. Error screening algorithms ensure data integrity and are also used for quality assurance. Customized output reports can be generated within minutes and have assisted in EMS quality assurance, planning, and research. We believe that the successful performance of this system supports the use of the suggested data elements as well as optical scanning and microcomputer analysis of EMS data.

Data Collection↗

MR of the knee: the significance of high signal in the meniscus that does not clearly extend to the surface.

On MR images of the knee it is sometimes impossible to determine with confidence if a focus of high signal in the meniscus is confined to the substance of the meniscus or if it extends to involve the surface. This is a critical differentiation because the latter represents meniscal tears that can be found and treated at arthroscopy, whereas the former represents degeneration, tears, or perhaps normal variants that cannot be detected or treated arthroscopically. We make an equivocal diagnosis of a tear when it is difficult to decide if signal in a meniscus involves the meniscal surface. We studied MR scans of the knee in 142 consecutive patients for the presence of such equivocal tears. Their prevalence was 14% (20/142); 17 were in the posterior horn of the lateral meniscus and three were in the posterior horn of the medial meniscus. In 13 cases with arthroscopy/arthrotomy correlation, no tears were found. In one of the 20 patients in whom the meniscus was removed during arthroplasty, histologic examination of the meniscus showed separation of collagen bundles, which was caused by meniscal degeneration confined to the substance of the meniscus. These results suggest that a meniscal tear is unlikely when MR scans show a focus of high signal in a meniscus that does not unequivocally extend to involve the surface of the meniscus.

Adolescent↗

Maternal endocrine and fetal metabolic responses to heat stress.

Heat-stressed pregnant ewes deliver intrauterine growth-retarded lambs. Selected maternal and fetal changes were investigated during acute heat stress in order to elucidate the mechanism for this growth retardation. Uterine blood flow decreased 20 to 30% in pregnant ewes during 1 degree C increases in core temperature. The decreases were accompanied by 60 and 100% increases in serum oxytocin and antidiuretic hormone, respectively. These effects were mimicked by salt loading or injections of antidiuretic hormone or oxytocin, suggesting a role for either or both hormones in regulating uterine blood flow during pregnancy. Chronically heat-stressed pregnant ewes were delivered by Caesarean section. Their fetuses were approximately 20% smaller than thermoneutral controls. Within each pair of heat-stressed twins, one fetus weighted one-third less than its litter mate. No difference in weights were observed within the control twins. The livers and brains of the heat-stressed fetuses were disproportionate in size. The livers from the small heat-stressed twins contained only one-half the protein of the controls and one-fourth the protein of their litter mates. Muscle protein was decreased in the heat-stressed fetuses, and liver and muscle glycogen were elevated as were liver arginase, glutamate-pyruvate transaminase and muscle creatinine. These results are consistent with the following hypothesis: heat stress stimulates the release of maternal antidiuretic hormone or oxytocin, which reduces uterine blood flow and causes a shift in fetal metabolism from anabolic to catabolic pathways; one fetus of heat-stressed twins is more severely affected than its litter mate.

Animals↗

Single-exposure dual-energy computed radiography: improved detection and processing.

Recent reports have emphasized the potential for dual-energy computed radiographic applications. An improved method for single-exposure material-selective imaging with a photostimulable phosphor computed radiography system was investigated. The essential elements of the technique are (a) prefiltration with gadolinium, which divides the incident broad-beam x-ray spectrum into low-energy and high-energy peaks; (b) a cassette consisting of four photostimulable phosphor plates that record images of increasing mean energies, with a computed energy separation of 23 keV from the front to the rear plate; (c) spatially dependent scatter and beam-hardening corrections; and (d) a noise-reduction algorithm based on noise correlations between bone-selective and soft-tissue-selective dual-energy images. These elements result in improved material cancellation and signal-to-noise ratio throughout the image.

Algorithms↗

Preparation of serum oxytocin and arginine vasopressin prior to radioimmunoassay: simultaneous extraction and separation on C18 Sep-Pak cartridges.

Oxytocin (OT) and arginine vasopressin (AVP) are often secreted in response to the same stimuli. The hormones are seldom assayed together, however, because of labor intensive sample preparation and the duplicate volumes required. A method has been developed for the simultaneous extraction and separation of OT and AVP from a single serum sample. The method is suited for sample preparation prior to radioimmunoassay (RIA) and reduces sample volume and processing time by 50%. Serum, supplemented with labeled and unlabeled OT and AVP, was adsorbed onto C18 (octadecasilyl-silica, ODS) Sep-Pak cartridges. After washing with phosphosaline and 3% aqueous acetone, OT was eluted with 98% aqueous acetone followed by AVP with 80% acidified (0.02 mol/L HCl) acetone. The recoveries, determined by radioactivity and RIA measurements, were 86 +/- 3% (OT) and 71 +/- 7% (AVP). Cross contamination was less than 10%. Sep-Paks extracted up to 100 pg/mL of the hormones from 10 mL of serum. The method was employed to measure OT and AVP in the pregnant ewe. Both hormones were elevated during salt-loading and dehydration and were decreased by carotid infusions of ethanol.

Animals↗

Effect of blood sampling and shipment to slaughter on plasma catecholamine concentrations in bulls.

The effect of venipuncture on blood serum catecholamine concentrations was studied in five yearling Red Angus bulls. Treatments consisted of sampling jugular vein blood via indwelling cannulas in a squeeze chute every 10 min for 1 h; sampling jugular vein blood from an indwelling catheter before, during and after venipuncture simulation; sampling jugular vein blood by venipuncture in a chute after shipment of 320 km to a slaughter plant; and sampling pooled blood from the neck during exsanguination at slaughter. Plasma epinephrine (E) and norepinephrine (NE) concentrations were determined by high performance liquid chromatography (HPLC) with electrochemical detection (ECD). Prevenipuncture NE and E in blood sampled from an indwelling catheter extended through a solid wall were 1.0 and .5 pmol/ml plasma, respectively. Plasma NE and E during venipuncture (1.6 and 1.1 pmol/ml, respectively) and immediately after cannulation in a squeeze chute (1.8 and 1.2 pmol/ml, respectively) were higher than prevenipuncture concentrations. Plasma NE and E, 1.9 and 1.2 pmol/ml plasma after shipment of 320 km, increased during exsanguination to 135.8 and 81.3 pmol/ml plasma, respectively. The NE and E in blood samples collected by venipuncture in a chute or box stall do not represent resting concentrations of these catecholamines.

Animals↗

Reversal of depressed miduterine arterial flow during hyperthermia-induced respiratory alkalosis.

The influence of ambient and arterial PCO2 on miduterine arterial flow of pregnant sheep acutely exposed to hot environments was investigated. Five mixed-breed ewes between 120 and 130 days of gestation were subjected to hot environments (increasing from thermoneutral 23 to 40 degrees C), and arterial blood pH, PCO2, and PO2 were determined at 5-min intervals. Respiratory rate, heart rate, rectal temperature, blood pressure, and miduterine arterial flow were continuously monitored prior to and during elevation of ambient air temperature. When miduterine arterial flow had decreased to 50% of thermoneutral control levels, ambient air CO2 was increased to 2.5%. Elevated ambient inspired CO2 caused a reversal in arterial pH and PCO2 to near thermoneutral levels. Miduterine arterial flow increased to 77% of the control levels following the elevated ambient PCO2 period. Respiratory rate also decreased when ambient CO2 was increased but remained 136% greater than the thermoneutral control level. All other parameters remained near their heat stress (40 degrees C) level during the elevation of ambient CO2. These data indicate that heat-stress-induced depression of miduterine arterial flow is vasoactively regulated, and cause-effect related to both arterial pH and PCO2, and thermoregulatory shunting of blood to heat-dissipating surfaces.

Alkalosis, Respiratory↗

Bupivacaine disposition in mother, fetus, and neonate after spinal anesthesia for cesarean section.

Uptake of bupivacaine from the subarachnoid space and its placental transfer were measured in six patients undergoing elective cesarean section. Maternal plasma levels (59 +/- 32 ng/ml) were only about 5% of those found in a comparable previous study of epidural anesthesia. Mean plasma umbilical venous bupivacaine levels (20.2 +/- 21 ng/ml) were 7% of those found after epidural anesthesia. Mean umbilical venous/maternal venous bupivacaine ratios were 0.34 +/- 0.12 and mean umbilical arterial/umbilical venous ratios were 0.81 +/- 0.30. No bupivacaine was detectable in neonatal plasma 24 hr after delivery. Neonatal urine had measurable levels of both bupivacaine and its inactive metabolite, 2,6-pipecolylxylidine (PPX), for at least 36 hr after delivery. The results demonstrate that bupivacaine crosses the placenta and reaches the fetus, but in very low amounts. This transplacental passage occurs despite injection of only small doses of a very highly protein bound drug into the subarachnoid space.

Anesthesia, Obstetrical↗

Lack of influence of cimetidine on bupivacaine levels during parturition.

The concurrent use of cimetidine as a prophylactic antacid and bupivacaine as a local anesthetic for epidural anesthesia for cesarean section has been promoted. However, cimetidine is known to inhibit clearance of many drugs by reducing hepatic blood flow and by inhibiting cytochrome P-450 enzymes. The purpose of this study was to determine whether cimetidine during epidural anesthesia alters the metabolism, disposition, protein binding, or placental transfer of bupivacaine. Thirty-six patients undergoing cesarean section with 0.5% bupivacaine for epidural anesthesia were studied. Sixteen patients received cimetidine (300 mg IM) 1-4 hr before cesarean section and 20 control patients received sodium citrate (30 ml PO) 10 min preoperatively. Bupivacaine and its inactive metabolite, 2,6-pipecolylxylidine (PPX) were quantitated by gas chromatography/mass spectrometry. Maternal plasma concentration time curves, fetal/maternal ratios at delivery, cord vein/cord artery ratios. PPX/bupivacaine ratios, and maternal and neonatal urinary excretion were compared between the two groups. No significant differences could be found between the groups in any of the outcome variables. However, in the presence of cimetidine, the percentage of unbound bupivacaine significantly increased from 1.36 +/- 0.63 to 1.66 +/- 0.78%. The results suggest that a single 300-mg IM dose of cimetidine does not significantly affect bupivacaine disposition or metabolism in parturients when given 1-4 hr before anesthesia.

Adult↗

Midcarpal instability.

The pathomechanics of midcarpal instability are described, particularly as they relate to the ring model of the wrist. A review of the clinical history, evaluation, and treatment of patients with midcarpal instability is presented.

Arthrodesis↗

Fed-batch culture technology.

The use of fed-batch procedures offers distinct advantages over other modes of operation of bioreactors, and is a widely researched technique. These advantages are discussed; some uses of fed-batch procedures and the associated methods of modelling and control are reviewed.

Journal Article↗

Ganglioside modulation of lipopolysaccharide-initiated complement activation.

Highly purified preparations of mouse gangliosides have been demonstrated to bind to purified preparations of lipopolysaccharide (LPS). In some instances, the binding has been demonstrated to be dependent upon the presence of sialic acid in the ganglioside preparation. The binding of gangliosides to LPS from the deep rough Salmonella minnesota Re mutant has suggested that the interaction involves the lipid A-2-keto-3-deoxyoctulosanate region of the LPS macromolecule. The interaction between gangliosides and LPS has been demonstrated to result in an abrogation of lipid A dependent activation of the classical pathway of serum complement by Re LPS. Surprisingly, however, the presence of sialic acid containing glycolipids has been shown to enhance significantly the capacity of LPS to initiate activation of the alternative pathway of complement. These data suggest that sialic acid can enhance as well as inhibit the formation of a stable alternative-pathway C3 convertase.

Animals↗

Lack of peripheral sympathetic control of uterine blood flow during acute heat stress.

Peripheral alpha- and beta-adrenergic control of uterine blood flow (UBF) during acute heat stress of the gravid ewe was investigated. An electromagnetic blood flow probe was surgically implanted around the left miduterine artery and catheters inserted in the left carotid artery and right jugular vein in ewes between d 120 and 130 of gestation. Four or more days postsurgery, ewes were fitted with instruments to measure rectal temperature (Tr), heart rate (HR), respiratory rate (RR), blood pressure (BP) and UBF. One-half hour after instrument calibration, a 15-min thermoneutral control period was initiated with carotid artery blood samples taken at 5-min intervals for pH and PCO2 determinations. Ewes were then subjected to a heat challenge that reached 40 C at 2 h. All physiological data were recorded every 5 min as 1-min mean values. In seven experiments on five ewes, an alpha-adrenergic blocking drug, phenoxybenzamine (PB) was infused at 1 mg/min for 15 min subsequent to maximum depression of UBF. A beta-adrenergic blocking drug, propranolol (PR) was infused at .35 mg/min for 15 min in eight experiments on five ewes. Analysis of variance comparisons were made between the control period and heat stress infusion periods within the PB and PR experiments. Further comparisons were made between the start and 5, 10 or 15 min of PB or PR infusion in order to test drug effects during an acute heat stress. Rectal temperature HR, RR and arterial pH were higher (P less than .05) at the start of PR and PB infusions than during the thermoneutral control period.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗