Biomedical subjects

A H Goldstein

Publications and source records attributed to A H Goldstein.

At least 19 recordsLinked to original sources

PubMed2001

Cloning of a Serratia marcescens DNA fragment that induces quinoprotein glucose dehydrogenase-mediated gluconic acid production in Escherichia coli in the presence of stationary phase Serratia marcescens.

Serratia marcescens ER2 was isolated from an endorhizosphere sample based on its high level of mineral phosphate solubilizing (MPS) activity. This phenotype was correlated with expression of the direct oxidation pathway. An ER2 plasmid library constructed in Escherichia coli strain DH5alpha was screened for MPS activity. A recombinant clone DH5alpha (pKG3791) was capable of gluconic acid (GA) production and tricalcium phosphate solubilization but only in the presence of stationary phase ER2 cells. GA production in DH5alpha (pKG3791) was apparently the result of the quinoprotein glucose dehydrogenase activity because AG121 (a Tn5 knockout of gcd) carrying pKG3791 did not produce GA under the same conditions. GA production by DH5alpha (pKG3791) was not observed when ER2 was replaced by another PQQ-producing strain bacterium. These data add to a growing body of evidence that E. coli contains some type of PQQ biosynthesis pathway distinct from those previously characterized in Gram-negative bacteria and that these genes may be induced under appropriate conditions.

Calcium Phosphates↗

PubMed2001

Large carbon isotope fractionation associated with oxidation of methyl halides by methylotrophic bacteria.

The largest biological fractionations of stable carbon isotopes observed in nature occur during production of methane by methanogenic archaea. These fractionations result in substantial (as much as approximately 70 per thousand) shifts in delta(13)C relative to the initial substrate. We now report that a stable carbon isotopic fractionation of comparable magnitude (up to 70 per thousand) occurs during oxidation of methyl halides by methylotrophic bacteria. We have demonstrated biological fractionation with whole cells of three methylotrophs (strain IMB-1, strain CC495, and strain MB2) and, to a lesser extent, with the purified cobalamin-dependent methyltransferase enzyme obtained from strain CC495. Thus, the genetic similarities recently reported between methylotrophs, and methanogens with respect to their pathways for C(1)-unit metabolism are also reflected in the carbon isotopic fractionations achieved by these organisms. We found that only part of the observed fractionation of carbon isotopes could be accounted for by the activity of the corrinoid methyltransferase enzyme, suggesting fractionation by enzymes further along the degradation pathway. These observations are of potential biogeochemical significance in the application of stable carbon isotope ratios to constrain the tropospheric budgets for the ozone-depleting halocarbons, methyl bromide and methyl chloride.

Bacteria↗

PubMed2001

Continuous flow stable isotope methods for study of delta(13)C fractionation during halomethane production and degradation.

Gas chromatography/mass spectrometry/isotope ratio mass spectrometry (GC/MS/IRMS) methods for delta(13)C measurement of the halomethanes CH(3)Cl, CH(3)Br, CH(3)I and methanethiol (CH(3)SH) during studies of their biological production, biological degradation, and abiotic reactions are presented. Optimisation of gas chromatographic parameters allowed the identification and quantification of CO(2), O(2), CH(3)Cl, CH(3)Br, CH(3)I and CH(3)SH from a single sample, and also the concurrent measurement of delta(13)C for each of the halomethanes and methanethiol. Precision of delta(13)C measurements for halomethane standards decreased (+/-0.3, +/-0.5 and +/-1.3 per thousand) with increasing mass (CH(3)Cl, CH(3)Br, CH(3)I, respectively). Given that carbon isotope effects during biological production, biological degradation and some chemical (abiotic) reactions can be as much as 100 per thousand, stable isotope analysis offers a precise method to study the global sources and sinks of these halogenated compounds that are of considerable importance to our understanding of stratospheric ozone destruction.

Biodegradation, Environmental↗

PubMed2001

Clinical results with the AB-180 left ventricular assist device.

BACKGROUND: This report reviews the initial clinical experience with the AB-180 ventricular assist device. METHODS: Between Dec 1997 and July 2000, the AB-180 was implanted in 17 patients at five institutions. The mean age was 52 years (range 21 to 68 years) and 14 of 17 were male. The indications for implantation were postcardiotomy shock (12 of 17, 70%), decompensated cardiomyopathy (2 of 17, 12%), viral myocarditis (2 of 17, 12%), and acute myocardial infarction (1 of 17, 6%). RESULTS: The mean duration of support was 8.5 days (range 1 to 28 days). In the group of 17 patients, 8 were weaned from the device and 2 underwent transplantation. Four of the weaned patients (4 of 8, 50%) and 1 of the transplant patients (1 of 2, 50%) survived. The overall weaning and survival rates were 58% (10 of 17) and 29% (5 of 17). There were no major device-related complications and no major device malfunctions. CONCLUSIONS: The AB-180 provides reliable circulatory support for reversible forms of heart failure.

Adult↗

PubMed2001

Carbon dioxide and water vapor exchange by young and old ponderosa pine ecosystems during a dry summer.

We investigated key factors controlling mass and energy exchange by a young (6-year-old) ponderosa pine (Pinus ponderosa Laws.) plantation on the west side of the Sierra Nevada Mountains and an old-growth ponderosa pine forest (mix of 45- and 250-year-old trees) on the east side of the Cascade Mountains, from June through September 1997. At both sites, we operated eddy covariance systems above the canopy to measure net ecosystem exchange of carbon dioxide and water vapor, and made concurrent meteorological and ecophysiological measurements. Our objective was to understand and compare the controls on ecosystem processes in these two forests. Precipitation is much higher in the young plantation than in the old-growth forest (1660 versus 550 mm year-1), although both forests experienced decreasing soil water availability and increasing vapor pressure deficits (D) as the summer of 1997 progressed. As a result, drought stress increased at both sites during this period, and changes in D strongly influenced ecosystem conductance and net carbon uptake. Ecosystem conductance for a given D was higher in the young pine plantation than in the old-growth forest, but decreased dramatically following several days of high D in late summer, possibly because of xylem cavitation. Net CO2 exchange generally decreased with conductance at both sites, although values were roughly twice as high at the young site. Simulations with the 3-PG model, which included the effect of tree age on fluxes, suggest that, during the fall through spring period, milder temperatures and ample water availability at the young site provide better conditions for photosynthesis than at the old pine site. Thus, over the long-term, the young site can carry more leaf area, and the climatic conditions between fall and spring offset the more severe limitations imposed by summer drought.

California↗

PubMed2001

Response of stomatal conductance to drought in ponderosa pine: implications for carbon and ozone uptake.

To gain insight into the limitations imposed by a typical Mediterranean-climate summer drought on the uptake of carbon and ozone in the ponderosa pine (Pinus ponderosa Dougl. ex Laws.) ecosystem, we compared diurnal trends in leaf physiology of young trees in a watered and a control plot located in the Sierra Nevada Mountains, CA, USA (Blodgett Forest, 38 degrees 53' N, 120 degrees 37' W, 1315 m elevation). Predawn water potential of trees in the watered plot remained above -0.3 MPa throughout the growing season, whereas it dropped in the control plot from -0.24 to -0.52 MPa between late May and mid-August. Photosynthesis and stomatal conductance of trees in the watered plot were relatively insensitive to atmospheric vapor pressure deficit (VPD), whereas gas exchange of trees in the control plot varied with changes in soil water, VPD and temperature. Although the 1998 growing season was abnormally wet, we saw a pronounced drought effect at the control site. Over the 2 months following the onset of watering, carbon and ozone uptake were measured on three days at widely spaced intervals. Carbon uptake per unit leaf area by 1-year-old foliage of trees in the control plot was 39, 35 and 30% less, respectively, than in the watered plot, and estimated ozone deposition per unit leaf area (ozone concentration times stomatal conductance) was 36, 46 and 41% less.

California↗

PubMed1999

An isotopic approach for understanding the CH(3)Br budget of the atmosphere.

The atmospheric budget of methyl bromide (CH(3)Br), an ozone-depleting gas, is highly uncertain, because it has complex sources and sinks. Although oceans, biomass burning, and industrial production are identified as the major sources, the fraction of CH(3)Br that is contributed by each source is not well known. A mass-balance approach that exploits differences in the carbon isotopic signature (delta(13)C) of CH(3)Br sources and sinks may provide a means of reducing uncertainties in the atmospheric budget. This approach depends on the distinctiveness of industrially produced methyl bromide. Our delta(13)C measurements of industrial CH(3)Br from the three largest manufacturers worldwide yield a weighted average of -54.4 per thousand relative to the Peedee Belemnite standard. This result suggests that industrial CH(3)Br is isotopically distinct and that the carbon isotopic composition of atmospheric CH(3)Br may indicate what fraction of atmospheric CH(3)Br is anthropogenic.

Air↗

PubMed1996

Small, low-cost implantable centrifugal pump for short-term circulatory assistance.

BACKGROUND: In 1991, Allegheny General Hospital and Allegheny-Singer Research Institute purchased a centrifugal pump, then a 2-year-old technology, from Medtronic Bio-Medicus, as part of its research program for novel treatments of acute and chronic heart failure. During a 4-year development program, we then established and met goals of durability, performance, thromboresistance, and low cost. METHODS: In vitro testing involved extensive hydraulic characterizations using Penn State mock loops. Calorimetry was used to determine efficiency. Durability studies used heated (37 degrees C) seawater for 28 to 45 days. In vivo studies used 46 sheep to test performance and engineering changes and to determine myocardial oxygen consumption, thromboresistance, and long-term durability. A left atrium-to-aorta circuit was used in all. RESULTS: Hydraulic testing showed no preload sensitivity but moderate afterload sensitivity at all impeller speeds (2,000 to 6,000 rpm). The heat load was low, and overall efficiency was 13% to 15%. Bench durability studies showed no electrical malfunction of the stator or console without degradation of the biomaterials used. Acute in vitro studies showed a near-linear relationship of myocardial oxygen consumption and left ventricular stroke work, pump flow, and pump speed. At speeds of 2 to 3 L/min (50% bypass), left ventricular stroke work and myocardial oxygen consumption were decreased approximately 50%. Additionally, 5 animals have had implants for 28 to 154 days with no macroemboli or microemboli detected in any animal. Hematologic and biochemical studies became normal 3 to 7 days after implantation. Hemolysis was low at less than 10 mg/dL. Clinical costs of the device are estimated to be 80% less than those of currently available devices. CONCLUSIONS: We conclude that an old technology has been made into new technology by application of sound engineering design principles, microchips, and new biomaterials. Qualifying trails for a Food and Drug Agency investigational device exemption application are in progress.

Animals↗

PubMed1995

Cloning of a mineral phosphate-solubilizing gene from Pseudomonas cepacia.

We have recently shown that the ability of some gram-negative bacteria to dissolve poorly soluble calcium phosphates (Mps+ phenotype) is the result of periplasmic oxidation of glucose to gluconic acid via the quinoprotein glucose dehydrogenase (GDH), a component of the direct oxidation pathway. Escherichia coli K-12 derivatives synthesize apo-GDH but not the cofactor pyrroloquinoline-quinone (PQQ) essential for formation of the holoenzyme. Therefore, in the absence of exogenous PQQ, these strains do not produce gluconic acid and are Mps-. Evidence is presented to show that expression of a single 396-base Pseudomonas cepacia open reading frame (designated gabY) in E. coli JM109 (a K-12 derivative) was sufficient to induce the Mps+ phenotype and production of gluconic acid. We present the nucleotide sequence of this open reading frame which coded for a protein (GabY) with a deduced M(r) of 14,235. Coupled transcription-translation of a plasmid (pSLY4 or pGAB1) carrying gabY resulted in production of a protein with an M(r) of 14,750. Disruption of the open reading frame of gabY via site-directed mutagenesis changed the phenotype to Mps- and eliminated gluconic acid production. The deduced amino acid sequence of gabY has no apparent homology with those of previously cloned direct oxidation pathway genes but does share regions highly homologous with the histidine permease system membrane-bound protein HisQ as well as other proteins in this family. In the presence of 1 microM exogenous PQQ, both JM109(pSLY4) and JM109(pGAB1) produced 10 times as much gluconic acid as was seen with either the plasmid or exogenous PQQ alone.(ABSTRACT TRUNCATED AT 250 WORDS)

ATP-Binding Cassette Transporters↗

PubMed1994

Predictable reduction in left ventricular stroke work and oxygen utilization with an implantable centrifugal pump.

Previous investigations with roller pumps and pneumatic pulsatile assist devices have demonstrated that nearly complete capture of normal left ventricular end-diastolic volume was necessary for appreciable reductions in oxygen consumption and stroke work. We tested the hypothesis that a centrifugal pump would decrease left ventricular stroke work and oxygen consumption as a function of pump flow. Ten sheep (35 to 50 kg) were instrumented and placed on left atrium-to-descending aorta bypass with a small, lightweight (112 g), implantable centrifugal pump. The relations between pump flow as a percent of cardiac output (% bypass), left ventricular stroke work, and oxygen consumption were studied. Left ventricular stroke work was calculated from the pressure-volume loops obtained with micromanometer and conductance catheters and was indexed per 100 g of left ventricular wet weight. Oxygen consumption was calculated from left main coronary artery blood flow and the arterial-coronary sinus oxygen content difference, normalized to 100 g of left ventricular wet weight and a heart rate of 100 beats/min. Measurements were made in stepwise increments of pump flow from zero to the maximum obtainable and then reversed in similar decrements. Analyses were made for 27 complete runs. Our data demonstrate that reductions in left ventricular stroke work and oxygen consumption were achieved from zero to maximal bypass. There was an approximate 66% and 50% reduction in left ventricular stroke work and oxygen consumption, respectively, at 60% bypass.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Cloning of an Erwinia herbicola gene necessary for gluconic acid production and enhanced mineral phosphate solubilization in Escherichia coli HB101: nucleotide sequence and probable involvement in biosynthesis of the coenzyme pyrroloquinoline quinone.

Escherichia coli is capable of synthesizing the apo-glucose dehydrogenase enzyme (GDH) but not the cofactor pyrroloquinoline quinone (PQQ), which is essential for formation of the holoenzyme. Therefore, in the absence of exogenous PQQ, E. coli does not produce gluconic acid. Evidence is presented to show that the expression of an Erwinia herbicola gene in E. coli HB101(pMCG898) resulted in the production of gluconic acid, which, in turn, implied PQQ biosynthesis. Transposon mutagenesis showed that the essential gene or locus was within a 1.8-kb region of a 4.5-kb insert of the plasmid pMCG898. This 1.8-kb region contained only one apparent open reading frame. In this paper, we present the nucleotide sequence of this open reading frame, a 1,134-bp DNA fragment coding for a protein with an M(r) of 42,160. The deduced sequence of this protein had a high degree of homology with that of gene III (M(r), 43,600) of a PQQ synthase gene complex from Acinetobacter calcoaceticus previously identified by Goosen et al. (J. Bacteriol. 171:447-455, 1989). In minicell analysis, pMCG898 encoded a protein with an M(r) of 41,000. These data indicate that E. coli HB101(pMCG898) produced the GDH-PQQ holoenzyme, which, in turn, catalyzed the oxidation of glucose to gluconic acid in the periplasmic space. As a result of the gluconic acid production, E. coli HB101(pMCG898) showed an enhanced mineral phosphate-solubilizing phenotype due to acid dissolution of the hydroxyapatite substrate.

Acinetobacter calcoaceticus↗

PubMed1992

Effects of crystalloid, blood, and University of Wisconsin perfusates on weight, water content, and left ventricular compliance in an edema-prone, isolated porcine heart model.

Coronary perfusion with blood and cardioplegic solutions was examined in isolated, arrested, hypothermic porcine hearts. Myocardial water content, heart weight, and left ventricular diastolic pressure-volume curves were measured before and after coronary perfusion. Statistics were based on exponential curve fitting to pressure-volume data and analysis of variance. Thirty-two pig hearts were divided into five experimental groups and a control group; after control measurements, each experimental group underwent three successive coronary perfusions with 1 L of unmodified blood or a solution of controlled osmolarity, 150 mOsm/L (diluted Plegisol solution), 280 mOsm/L (Plegisol solution and albumin), 334 mOsm/L (University of Wisconsin solution), or 380 mOsm/L (Stanford solution). After each perfusion, measurements were repeated. All experiments were completed within 90 minutes. The first perfusion was delayed 20 minutes after excision of the heart to allow for instrumentation. Each experimental group demonstrated a statistically significant increase in heart weight and myocardial water content and a significant decrease in left ventricular compliance after perfusion. Changes were less pronounced with blood than crystalloids. Edema effects were minimized but not prevented by hyperosmolarity. University of Wisconsin solution appeared unique in minimizing progressive edema after the first perfusion. Over the 81 perfusions studied, changes in left ventricular compliance were linearly related to heart weight and water content. We conclude that in this model, in which edema sensitivity is increased by delayed perfusion and venous occlusion, edema is minimized but not eliminated by whole blood and University of Wisconsin solution. The model appears useful in assessing properties of cardioplegia vehicles intended for use in the injured myocardium.

Adenosine↗

PubMed1989

Phosphate Starvation Inducible Metabolism in Lycopersicon esculentum: III. Changes in Protein Secretion under Nutrient Stress.

Phosphate starvation increased the secretion of at least six proteins by suspension cultured tomato (Lycopersicon esculentum L. and L. pennellii) cells. Cells exhibited a biphasic response to phosphate (Pi) starvation. The early phase involved enhanced secretion of three proteins in response to transfer to a Pi-depleted media, while biomass accumulation continued at the same rate as in the Pi-sufficient cells. Severe starvation, defined as inhibition of biomass accumulation, induced enhanced secretion of three additional proteins. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis, media proteins were immunoblotted with antibodies reacting specifically to oligosaccharides processed by the Golgi apparatus. Binding patterns showed that the enhancement in secretion during both phases of starvation was Golgi-mediated. Cells undergoing severe starvation had a respiration rate approximately twice that of unstressed cells and secreted 4.4 times more protein into the media per unit biomass. These data suggest overlapping Pi starvation-specific and global stress responses in plant cells. Under these conditions, Golgi-mediated protein secretion is enhanced. We present evidence for phosphate starvation inducible enhancement of Pi uptake. Secreted proteins specific for N and Fe starvation are also identified.

Journal Article↗

PubMed1988

Phosphate Starvation Inducible Metabolism in Lycopersicon esculentum: I. Excretion of Acid Phosphatase by Tomato Plants and Suspension-Cultured Cells.

Both tomato (Lycopersicon esculentum cv VF 36) plants and suspension cultured cells show phosphate starvation inducible (psi) excretion of acid phosphatase (Apase). Apase excretion in vitro was proportional to the level of exogenous orthophosphate (Pi). Intracellular Apase activity remained the same in both Pi-starved and sufficient cells, while Apase excreted by the starved cells increased by as much as six times over unstressed control cells on a dry weight basis. At peak induction, 50% of total Apase was excreted. Ten day old tomato seedlings grown without Pi showed slight growth reduction versus unstressed control plants. The Pi-depleted roots showed psi enhancement of Apase activity. Severely starved seedlings (17 days) reached only one-third of the biomass of unstressed control plants but, because of a combination of psi Apase excretion by roots and a shift in biomass to this organ, they excreted 5.5 times the Apase activity of the unstressed control. Observed psi Apase excretion may be part of a phosphate starvation rescue system in plants. The utility of the visible indicator dye 5-bromo-4-chloro-3-indolyl-phosphate-p-toluidine as a phenotypic marker for plant Apase excretion is demonstrated.

Journal Article↗

PubMed1988

Phosphate Starvation Inducible Metabolism in Lycopersicon esculentum: II. Characterization of the Phosphate Starvation Inducible-Excreted Acid Phosphatase.

Three-day-old suspension cultured cells of Lycopersicon esculentum transferred to a Pi-depleted medium had 2.7 times the excreted acid phosphatase (Apase) activity of cells transferred to a Pi-sufficient medium. Cell growth during this time period was identical for the two treatments. Excreted Apase activity was resolved into two fractions on a Sephadex G-150 column. Most of the phosphate starvation inducible (psi) enhancement in activity was in the lower molecular weight fraction. These two fractions exhibited different substrate versus pH activity profiles. With a native polyacrylamide gel electrophoresis assay, the lower molecular weight fraction resolved into two bands of activity. Both column fractions resolved into the same single band of activity with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The apparent molecular weight of this enzyme was 57 kilodalton. These data indicate that L. esculentum has at least two isozymes of the psi-excreted Apase and that these isozymes may associate to form high molecular weight aggregates. Labeling studies using [(35)S]methionine show that the psi response in tomato cells is complex and involves changes in the steady state levels of several excreted proteins.

Journal Article↗

PubMed1985

Phosphate Transport across the Plasma Membrane of Wheat Leaf Protoplasts: Characteristics and Inhibitor Specificities.

The kinetics and inhibitor specificities of phosphate transport across the plasma membrane of wheat leaf mesophyll protoplasts have been examined. Studies were also carried out on the effects of light and pH on phosphate transport and the plasma membrane electropotential. At pH 5.8 (30 degrees C), protoplasts accumulated phosphate at the rate of 3.9 +/- 0.2 nanomoles per milligram protein per hour. Phosphate uptake rates and inhibitor specificities for the leaf cell plasma membrane phosphate transporter were qualitatively similar to those observed with root protoplasts. Neither picrylsulfonic acid, or p-chloromercuribenzene sulfonate affected phosphate uptake significantly at 0.1 millimolar. Of all compounds tested, carbonyl cyanide-p-trifluoromethoxy phenylhydrazone was the most effective inhibitor of phosphate uptake (60% at 0.1 millimolar). Tribenzylphosphate inhibited uptake by 34% while dibenzylphosphate had no effect. The plasma membrane electropotential was found to be -37 +/- 3 millivolts. Initiation of photosynthesis lowered the membrane potential to -39 +/- 3 millivolts. Inhibition of phosphate uptake by 34% with the substrate analog tribenzylphosphate resulted in a measured membrane potential of -33 +/- 3 millivolts. These changes in potential were not significant at the 5% probability level. Phosphate uptake rates remained constant under photosynthetic and nonphotosynthetic conditions. The utility of tribenzylphosphate as an inhibitor in plant systems is demonstrated.

Journal Article↗

PubMed1981

Effects of protectants on the activity of wheat mitochondria.

A mitochondrial fraction was isolated from durum wheat seedlings via differential centrifugation using three different tissue grinding buffers. One buffer contained bovine serum albumin (BSA), a protein proven effective in protecting mitochondria from damage during isolation. The other two buffers contained either polyethyleneglycol of M.W. 20,000D (PEG-20), a phenolic binding compound or PEG-20 plus XAD-2, a non-ionic adsorbent material. Organelles isolated with the BSA exhibited an average ADP/O ratio of 2.3 and an average respiratory control ratio of 5.1, substantially higher than those obtained with the other media. Addition of the XAD-2 to a buffer containing PEG-20 resulted in an increase in the respiratory control ratio to an average value of 3.7 vs 2.7 for organelles isolated with PEG-20 alone without causing a concomitant rise in the ADP/O ratio which remained an average of 1.9. Protein values for the mitochondrial fraction isolated with BSA ranged from 2.5 to 3 times greater than those obtained from the PEG-20 preparations. This difference in organelle protein is discussed in relation to the accurate measurement of mitochondrial respiration.

Cell Fractionation↗

PubMed1981

Cyanide-insensitive and Cyanide-sensitive O(2) Uptake in Wheat: II. GRADIENT-PURIFIED MITOCHONDRIA LACK CYANIDE-INSENSITIVE RESPIRATION.

Enriched mitochondrial fractions isolated from durum wheat seedlings via differential centrifugation exhibited classical cyanide- or antimycin A-insensitive O(2) uptake which was inhibited by either salicylhydroxamic acid or propyl gallate. Further purification of this fraction using Percoll density gradients resulted in two discrete bands which were essentially homogeneous mitochondrial populations, as verified by electron microscopy. Respiratory O(2) uptake in these two fractions was completely inhibited by cyanide or antimycin A. Addition of linoleic acid to a third-step gradient band, which was shown to contain virtually no mitochondria, resulted in demonstrable cyanide-insensitive O(2) uptake. This O(2) consumption was completely inhibited by propyl gallate or salicylhydroxamic acid, two known lipoxygenase inhibitors. In contrast, addition of linoleic acid to the two purified mitochondrial fractions did not stimulate O(2) uptake. These data indicate that lipoxygenase oxygenation, the enzyme physically separable from the mitochondria, is responsible for the cyanide-insensitive component of O(2) uptake that was observed in subcellular fractions isolated from etiolated wheat seedlings.

Journal Article↗