Effects of fractions from biodegraded Alaska North Slope crude oil on embryonic inland silversides, Menidia beryllina.

Embryonic inland silversides, Meinida beryllina, were exposed to neutral, water-soluble fractions (WSFs) resulting from microbial degradation of artificially weathered Alaska North Slope (ANS) crude oil. Three individual microbes obtained from Prince William Sound, Alaska, and designated Phe#6 (enriched on phenanthrene), Hexaco#2 (enriched on the straight-chain alkane, hexacosane), and EI2V (grown by enrichment on Bushnell-Haas medium containing 0.2% pristane, a branched alkane) were used to individually biodegrade weathered ANS crude oil for 14 days in darkness in 20-L glass carboys containing nutrient enriched, sterilized 20% salinity sea water at 20 +/- 1 degrees C. Neutral WSFs resulting from biodegradation of ANS (lot 521) by each microbe were recovered and weighted. Neutral WSFs recovered were: 1.76 mg/L for Phe#6, 1.85 mg/L for Hexaco#2, and 13.02 mg/L for the EI2V microbe. Embryo toxicity and teratogenicity tests revealed that exposure of embryos to the WSFs from the EI2V incubation (with a total recovered neutral fraction approximately seven times greater than the Phe#6 and Hexaco#2 incubations) resulted in the most severe responses in craniofacial, cardiovascular, and skeletal organ systems. The total neutral WSFs recovered from the EI2V biodegradation of weathered ANS 521 were subfractionated into saturated (eluted with hexane), aromatic (eluted with CH2Cl2), polar (eluted with ethyl ether), and recombined (saturated + aromatic + polar) fractions. Developing fish embryos were then exposed to each subfraction and the recombined subfractions. The polar subfraction and recombined subfractions proved to be the most embryo toxic and teratogenic. They resulted in statistically significant (p < or = 0.05) responses (compared to controls) for craniofacial, cardiovascular, skeletal, and total severity effects in one or both tests with these subfractions.

Impact of abciximab and coronary stenting on outcomes and costs of percutaneous coronary interventions in a community hospital.

OBJECTIVE: To assess costs and outcomes of coronary stenting and balloon angioplasty with and without adjunctive treatment with abciximab for 3758 consecutive elective percutaneous coronary interventions at a single community center over the 2.5-year period between 1 January 1995 and 30 June 1997. RESULTS: Abciximab was more common among patients who had recently suffered myocardial infarction, patients with unstable angina, and patients with more complex coronary lesions. Use of abciximab in conjunction with balloon angioplasty or stenting and stenting alone was associated with significant reductions in incidence of major adverse cardiovascular events in hospital. Multivariate analysis indicated that use of abciximab and stenting were associated with significant independent effects on risk of an event. Hospital costs were increased for patients administered abciximab, treated with stenting, or both. Total costs and costs inclusive of those incurred in catheterization laboratory and pharmacy increased significantly with increasing complexity of lesions. Multivariate regression analysis (baseline cost US$5621) identified death (US$16098), emergency revascularization (US$13678), usage of multiple stents (US$1423 for each stent), and use of abciximab (US$1269) as independent predictors of a greater cost. One-year follow-up revealed significant differences among treatment strategies in terms of risk of need for subsequent revascularization procedures. Lack of stenting but not use of abciximab was identified as a significant predictor of need for repeat revascularization procedures. CONCLUSIONS: Our findings are in general agreement with cost analyses of use of abciximab for populations in clinical trials and suggest that improvements of early clinical outcome with abciximab treatment and stenting justify the incremental cost of treatment in a community hospital setting.

Degradation of weathered oil by mixed marine bacteria and the toxicity of accumulated water-soluble material to two marine crustacea.

Artificially weathered crude oil was degraded by four diverse cultures of mixed marine bacteria under optimized conditions for 7 and 14 days. Loss in total weight of starting oil (30 g) ranged from 6.8-17.3% in biologically active incubations compared with only 0. 9-1.1% in sterile and nutrient-limited controls. In all incubations, both neutral and acidic water-soluble fractions (WSF) were accumulated. In biologically active systems, 50.9-249.0 mg neutral and 63.3-406.8 mg acidic WSF were accumulated whereas only 6.5-11.1 mg neutral and 1.7-2.2 mg acidic WSF were accumulated in control incubations. Analysis by gas chromatography demonstrated that accumulated WSF in biologically active systems contained compounds different from those washed from the starting crude oil. Exposure of grass shrimp (Palaemonetes pugio) embryos to neutral WSF from each of the biologically active cultures resulted in high embryo mortalities relative to sterile and nutrient-limited controls which exhibited >90% hatching success and larval survival. Toxicity of neutral WSF was also demonstrated on larvae of mysids (Mysidopsis bahia). In both cases, toxicity occurred only on exposure to neutral material accumulated by active, oil-degrading cultures and not with material washed from the weathered crude oil. These results imply that unique compounds were accumulated during degradation that may have been responsible for increased toxicity.

Preliminary observations on responses of embryonic and larval Pacific herring, Clupea pallasi, to neutral fraction biodegradation products of weathered Alaska North Slope oil.

Weathered Alaska North Slope crude oil (ANS 521) was subjected to biodegradation in vigorously stirred incubations for 14 days at 15 +/- 1 degrees C in 20/1000 salinity sterilized seawater, amended with nutrients and inoculated with a hydrocarbon-degrading microorganism (EI2V) isolated from an oil-contaminated beach in Prince William Sound, Alaska. A total of 13.7 mg/L water-soluble neutral fraction (WSF) was recovered from the incubation of weathered ANS 521. Toxicity/ teratogenicity tests were conducted with WSF recovered from the biodegradation system using embryonic and larval Pacific herring, Clupea pallasi. Exposures were begun at 4, 48, and 96 h postfertilization of herring eggs. Exposure concentrations were 1, 10, and 100% of the original concentration of WSF recovered from incubations (redissolved in 20/1000 salinity sterile seawater at 15 +/- 1 degrees C). Sterile 20/1000 salinity seawater without the addition of redissolved neutral fraction was used as a control. Significant (p < or = 0.05) embryo mortality or teratogenic responses were observed at WSF concentrations of 10 and 100%. On days 5 through 8 of embryogenesis, counts of heart contraction rates were significantly lower (p < or = 0.05) at the 100% WSF concentration for embryos exposed beginning at 4 and 48 h postfertilization. Grow-out of larvae from selected exposures was conducted. High mortality was noted in larvae exposed to the 10% WSF concentration beginning at 4 and 48 h postfertilization. Most of these larvae died 5 to 8 days after hatching when they elicited vertebral displacements at a time concurrent with the onset of feeding behavior.

Modified heparin-bonded catheter for cannulation of the coronary sinus from the femoral vein.

Serial sampling from the coronary sinus is an attractive technique for drawing blood samples to be used in the characterization of procoagulant activity during coronary interventions. We have developed a modified Simmons catheter for rapid cannulation of the coronary sinus from the femoral approach. The catheter design incorporates heparin bonding and distal side-holes to minimize blood sampling artifacts. Coronary sinus cannulation was performed via the femoral approach in 186 patients by use of a multipurpose catheter (n = 8), an unmodified Simmons I or II catheter (n = 64), or the modified Simmons catheter (n = 114). The coronary sinus was cannulated successfully with the modified Simmons catheter in 97% of patients; the success rate with the unmodified Simmons II catheter was 87% (P = 0.02). The modified Simmons catheter represents an improved technique for cannulation of the coronary sinus from the femoral vein.

Association of heparin-resistant thrombin activity with acute ischemic complications of coronary interventions.

BACKGROUND: Acute thrombosis is thought to contribute to abrupt coronary occlusion during percutaneous coronary revascularization despite the administration of heparin and aspirin. This study was designed to detect the presence of heparin-resistant thrombin activity and to define its relationship to the acute ischemic complications of coronary interventions. METHODS AND RESULTS: Plasma levels of fibrinopeptide A (FPA) and prothrombin fragment 1.2 (F1.2), markers of thrombin and factor Xa activity, respectively, were measured in the coronary sinus with heparin-bonded catheters in 58 patients undergoing coronary interventions. Activated coagulation times were maintained > 300 seconds by the Hemochron method. Mean FPA levels decreased significantly, from 7.0 +/- 0.9 nmol/L before the procedure to 5.2 +/- 0.5 nmol/L after the heparin bolus and to 2.9 +/- 0.2 nmol/L after the procedure (P = .0001). In 26 patients (45%), FPA levels remained above the threshold for suppression angioplasty of thrombin activity determined during angiography in 7 patients without coronary artery disease (> 3.0 nmol/L). FPA concentrations after coronary interventions were increased in patients with intracoronary thrombus (P = .01), abrupt coronary occlusion (P = .06), postprocedural non-Q-wave myocardial infarction (P = .04), and clinically unsuccessful procedures (P = .04). F1.2 levels were relatively low before the procedures and did not change significantly. CONCLUSIONS: Heparin administration suppresses thrombin activity in most but not all patients undergoing coronary interventions. Heparin-resistant thrombin activity is associated with angiographic evidence of intracoronary thrombus and ischemic complications of coronary interventions.

Responses of embryonic and larval inland silversides, Menidia beryllina, to a water-soluble fraction formed during biodegradation of artificially weathered Alaska North Slope crude oil.

Weathered Alaska North Slope crude oil (ANS 521) was stirred for 2 and 14 days in 20 per thousand salinity sterile seawater or for 14 days in seawater with nutrients and a group of three (GO3) microorganisms from Prince William Sound, Alaska, that were capable of biodegrading hydrocarbons. A total of 0.65 and 0.69 mg/L water soluble fraction (WSF) of neutral fraction hydrocarbons was recovered from the 2- and 14-day stirred sterile systems, respectively. In comparison, a total of 7.5 mg/L WSF neutral fraction hydrocarbons was recovered from systems containing ANS 521 that were stirred and biodegraded by the GO3 microbes for 14 days. Toxicity/teratogenicity tests were conducted with neutral fraction hydrocarbons recovered from the sterile and biodegraded systems using embryonic inland silversides, Menidia beryllina. Hydrocarbons from the sterile systems did not cause statistically significant teratogenic responses at concentrations of 1%, 10%, and 100% (w/v) of recovered fractions (redissolved in 20 per thousand salinity sterile seawater). Counts of heart contraction rates were significantly lower (alpha

Usefulness of dobutamine stress echocardiography for the prospective identification of the physiologic significance of coronary narrowings of moderate severity in patients undergoing evaluation for percutaneous transluminal coronary angioplasty.

Dobutamine stress echocardiography (DSE) was performed after coronary angiography to evaluate the need to perform percutaneous transluminal coronary angioplasty (PTCA) for 46 stenoses of moderate severity (50% to 80%) in 46 patients. Patients were divided into 2 groups according to the DSE results in the distribution of the coronary artery with the lesion of moderate severity: group I (n = 32) were those without inducible myocardial ischemia; PTCA was not performed. Group II (n = 14) were those who exhibited myocardial ischemia; PTCA was performed in 12. The 2 groups were comparable in terms of clinical characteristics. Follow-up DSE was performed < or = 48 hours after PTCA, at 3 months, and 6 to 12 months after the first DSE. In group I at 3 months, DSE results were still negative in the distribution of the vessel with the moderately severe lesion in 24 patients; only 1 patient had a positive result, and 8 patients who refused DSE remained clinically stable. At 6 to 12 months (mean 7 +/- 2), 26 patients had negative study results; 3 patients who refused follow-up DSE remained clinically stable. In group II, 12 of 14 patients with inducible ischemia on the initial DSE underwent PTCA. Early follow-up DSE (< or = 48 hours) was negative in 7, and 4 had persistent inducible wall motion abnormalities in the myocardium subtended by the coronary artery in which the PTCA had been performed; 1 study was not performed.(ABSTRACT TRUNCATED AT 250 WORDS)

The Drosophila molybdenum cofactor gene cinnamon is homologous to three Escherichia coli cofactor proteins and to the rat protein gephyrin.

Essentially all organisms depend upon molybdenum oxidoreductases which require a molybdopterin cofactor for catalytic activity. Mutations resulting in a lack of the cofactor show a pleiotropic loss of molybdoenzyme activities and thereby define genes involved in cofactor biosynthesis or utilization. In prokaryotes, two operons are directly associated with biosynthesis of the pterin moiety and its side chain while additional loci play a role in the acquisition of molybdenum and/or activation of the cofactor. Here we report the cloning of cinnamon, a Drosophila molybdenum cofactor gene encoding a protein with sequence similarity to three of the prokaryotic cofactor proteins. In addition, the Drosophila cinnamon protein is homologous to gephyrin, a protein isolated from the rat central nervous system. Our results suggest that some portions of the prokaryotic cofactor biosynthetic pathway composed of monofunctional proteins have evolved into a multifunctional protein in higher eukaryotes.

Pseudoaneurysm following successful excimer laser coronary angioplasty of a restenotic left internal mammary artery graft ostial lesion.

Although significant left internal mammary artery graft ostial stenosis is extremely rare, the clinical importance can be profound. In this report we describe a case in which a restenotic left internal mammary artery graft ostial lesion was successfully opened with excimer laser coronary angioplasty. A resulting pseudoaneurysm spontaneously closed after conservative therapy.

Concordance of nutritive myocardial perfusion reserve and flow velocity reserve in conductance vessels in patients with chest pain with angiographically normal coronary arteries.

We have previously shown that myocardial perfusion can be quantified by positron emission tomography (PET) with 15O-labeled water (H2(15)O), as experimentally validated with radiolabeled microspheres in animal hearts. The purpose of our study was to determine whether myocardial nutritive perfusion reserve assessed with PET in human subjects was parallel to flow velocity reserve assessed in conductance vessels measured with intracoronary Doppler probes. We studied nine patients with chest pain and angiographically normal coronary arteries with intracoronary Doppler flow velocity assessments before and after administration of 16 micrograms of intracoronary adenosine. We also assessed myocardial nutritive perfusion with PET and H2(15)O before and after intravenous administration of dipyridamole (0.56 mg/kg). Perfusion reserve (the ratio of absolute values of myocardial perfusion after dipyridamole administration to perfusion at rest) estimated with PET (3.5 +/- 0.9 s.d.) correlated closely with flow velocity reserve (the ratio of hyperemic intracoronary flow velocity to flow velocity at rest) (3.5 +/- 1.2, r = 0.80, p < 0.01). Absolute values of perfusion assessed tomographically averaged 1.22 +/- 0.19 ml/g/min in patients at rest and 4.16 +/- 0.93 after dipyridamole administration. Our data indicate that noninvasive assessment of myocardial perfusion with PET provides results that parallel intracoronary Doppler flow velocity measurements. Because PET delineates nutritive perfusion throughout the heart in absolute terms, its use may facilitate detection of impaired coronary arterial function and enhance delineation of the efficacy of potentially therapeutic interventions in patients with chest pain and angiographically normal coronary arteries.

Implementation and evaluation of a two-compartment model for quantification of myocardial perfusion with rubidium-82 and positron emission tomography.

Positron emission tomography offers the ability to noninvasively assess regional myocardial perfusion in absolute terms (i.e., milliliters per gram per minute). Accurate estimates have been difficult to achieve with generator-produced 82Rb because of the complex behavior of this tracer in the myocardium. The aim of the present study was to determine whether regional myocardial blood flow could be assessed quantitatively with 82Rb and positron emission tomography by using a two-compartment kinetic model. Regional perfusion in milliliters per gram per minute was estimated from dynamic tomographic scans after intravenous administration of 82Rb in 18 studies in 13 intact dogs studied without intervention, after 2 and 24 hours of induced ischemia, during reperfusion after transient occlusion, or at rest and after pharmacological hyperemia after induced coronary artery stenosis. Regional flow was estimated along with the forward and backward rates of transport (k1 and k2 [minutes-1]) after the relative volume of distribution of the first compartment was fixed to 0.53 ml/ml and the tomographic parameters, the recovery and spillover fractions, were fixed to averaged values obtained in previous studies. In 36 comparisons, estimates of regional flow with 82Rb correlated well with flow measured with concomitantly administered radiolabeled microspheres (r = 0.91, p less than 0.05) over the flow range from 0.14 to 4.25 ml/g/min. A putative index of viability, k2, increased significantly in regions with severe ischemia. The results suggest that quantification of regional myocardial perfusion is possible in centers using 82Rb for estimates of myocardial perfusion when a physiologically appropriate, two-compartment model is used.

Noninvasive quantification of regional myocardial perfusion with rubidium-82 and positron emission tomography. Exploration of a mathematical model.

Positron emission tomography (PET) centers without cyclotrons use generator-produced rubidium-82 (82Rb) for assessment of myocardial perfusion. The aim of the present study was to determine whether myocardial blood flow could be assessed quantitatively with 82Rb and PET. Because the myocardial extraction fraction of 82Rb varies inversely and nonlinearly with flow and cannot be measured conveniently with PET, we used an experimentally derived mathematical function defining the relation between single-pass extraction fraction of 82Rb and flow to obviate the necessity of measuring the extraction fraction directly. Myocardial blood flow in absolute terms (ml/g/min) was estimated from dynamic PET scans after intravenous administration of 82Rb in intact dogs and compared with flows measured with radiolabeled microspheres. In 36 comparisons in 13 dogs studied at rest, or after coronary occlusion, reperfusion, or after coronary hyperemia induced with intravenous dipyridamole, over the flow range from 0.2 to 2.0 ml/g/min, estimates of perfusion with rubidium correlated well with flows measured concomitantly with microspheres, although there was a slight underestimation of flow with rubidium (flow by 82Rb = 0.92 x flow by microspheres-0.021, r = 0.83). In general, estimates of flow in ischemic regions were less reliable than estimates for regions with normal flow. Thus, although the relation between myocardial extraction and retention of 82Rb and flow can vary under a variety of physiological and pathophysiological conditions, this study demonstrates the ability to obtain quantitative estimates of myocardial blood flow with 82Rb and PET under carefully defined conditions without measuring the extraction fraction directly.

Assessment of regional myocardial and renal blood flow with copper-PTSM and positron emission tomography.

We recently demonstrated in isolated, perfused hearts that radiolabeled pyruvaldehyde bis(N4-methylthiosemicarbazonato)copper(II) (Cu-PTSM) is well extracted throughout a range of conditions including ischemia, hypoxia, and hyperemia. Once extracted, binding of radioactivity by the isolated heart was essentially irreversible, giving this tracer microspherelike qualities. Because Cu-PTSM can be readily prepared with the generator-produced positron-emitting copper 62 and other gamma- or positron-emitting copper radionuclides, we evaluated its usefulness for measuring regional myocardial and renal blood flow in vivo in intact dogs at rest, after ischemia, or after coronary hyperemia was induced by intravenous administration of dipyridamole. After intravenous administration of radiolabeled Cu-PTSM, the tracer cleared rapidly from the blood. Myocardial uptake of single photon-emitting 67Cu-labeled Cu-PTSM was measured directly in myocardial samples 15 minutes after tracer administration, and it increased proportionally with blood flow throughout the flow range (estimated concomitantly with radiolabeled microspheres) of 0.0-6.0 ml/g/min (n = 340 samples from 17 dogs, r = 0.99, Ycopper radioactivity = 85Xmicrosphere flow -7 chi 2 + 17). Renal uptake of radiolabeled Cu-PTSM was also proportional to blood flow. Positron emission tomography was performed in four intact dogs after intravenous administration of 64Cu-labeled Cu-PTSM (19% positron decay, t1/2 = 12.8 hours). High-quality images of heart and kidney were obtained. Accordingly, radiolabeled Cu-PTSM should be a useful, generator-produced tracer for estimating regional myocardial and renal blood flow with positron emission tomography.

In vivo delineation of myocardial hypoxia during coronary occlusion using fluorine-18 fluoromisonidazole and positron emission tomography: a potential approach for identification of jeopardized myocardium.

Previous studies have demonstrated that the positron-emitting fluorine-18 (18F)-labeled fluoromisonidazole is a specific tracer of myocardial hypoxia. Its fractional extraction is enhanced in ischemic or hypoxic myocardium but returns to baseline levels on reperfusion and recovery of normal function. Thus, this agent might be useful in delineating acutely hypoxic but potentially salvageable myocardium. Accordingly, to delineate the relation between the myocardial extraction of 18F-fluoromisonidazole after intravenous administration and the time of antecedent ischemia in vivo, uptake of tracer was measured with positron emission tomography and direct postmortem tissue analysis in 14 dogs in which tracer was administered within 3 h of coronary occlusion (a time associated with marked potential for salvage on reperfusion); in 4 dogs after 6 h of coronary occlusion (a time associated with minimal salvage of myocardium on reperfusion); and in 8 dogs after greater than 24 h of coronary occlusion (to delineate uptake in tissue that is irreversibly damaged). The residual fraction (that is, the amount of tracer extracted and retained in a region) in ischemic myocardium in the dogs in which 18F-fluoromisonidazole was administered within 3 h after occlusion averaged (+/- standard deviation) 23 +/- 18%, which was higher than the residual fraction in myocardium subjected to ischemia for either 6 or greater than 24 h before tracer administration (12 +/- 7% and 5 +/- 2%, respectively, p less than 0.01 for both). Retention of tracer in remote normal myocardium averaged 2 +/- 1%.(ABSTRACT TRUNCATED AT 250 WORDS)