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PubMed1995

Acute trauma alters morphine clearance.

Trauma is accompanied by changes in liver perfusion and acute phase proteins. Such changes have the potential to alter drug metabolism. There are few studies describing drug disposition in acute trauma. We determined the pharmacokinetics of an intermediate extraction drug, morphine, in trauma patients. Nine patients with an Injury Severity Score (ISS) > or = 16 were studied within 48 hours of trauma. Morphine 5 mg was given intravenously and serial blood and urine samples were drawn to derive pharmacokinetic parameters. Alpha acid glycoprotein (AAG) levels were determined. Total morphine clearance (CL) and volume of distribution (Vss) were decreased compared to established literature values. Area under the curve (AUC) and terminal half-life (T 1/2 alpha) were increased. AAG levels were higher than reference range. Elimination half-life was increased. The decrease we observed in Vss may be attributed to increased binding of morphine by AAG, which is increased after trauma as in our patients. Decreased clearance and increased half-life of an intermediate extraction drug may be explained by increased protein binding, decreased liver blood flow, and reduced hepatocellular function. Decreased clearance of the magnitude observed in these patients could result in drug accumulation. Better understanding of the effects of trauma on the pharmacokinetics of low, high, and intermediate extraction drugs will prevent excessive or suboptimal drug dosing.

Acute Disease↗
PubMed1998

Assessment of the effectiveness of composting for the reduction of toxicity and mutagenicity of explosive-contaminated soil.

Composting is being developed as an economical method for remediating explosive-contaminated soils and has been found to reduce the concentrations of target contaminants such as 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). However, whether environmental safety is improved by composting can be determined only by assessing the effects of the treated material on living organisms. In this study two bioassays, the Mutatox assay and the earthworm acute toxicity test, were used to evaluate the effectiveness of a pilotscale composting demonstration in reducing environmental hazard. Explosive-contaminated soil was collected from a military installation and amended for composting in two adiabatic reactors. The unamended soil was lethal to all exposed earthworms, as were both amended replicates, prior to composting. Serial dilutions of the finished composts with artificial soil had earthworm 14-day LC50 values of 35.7 and 100% finished compost: artificial soil. Extracts of the initial materials were also toxic to bacteria in the Mutatox assay. Dilutions of those extracts to sublethal concentrations revealed a low level of mutagenicity. Extracts of the finished composts indicated reduced bacterial toxicity, but the mutagenicity was markedly increased by composting. The reduction in lethality reflected the attenuation of explosives caused by composting, as indicated by chemical analysis. However, the increased mutagenicity was a result that would not have been indicated by chemical analysis alone and is inferred to be the result of the formation of mutagenic metabolites of explosives during composting and their incomplete degradation.

Animals↗
PubMed1995

Effect of histamine on endothelial permeability and structure and adhesion molecules of the paracellular junctions of perfused human placental microvessels.

The microvessels of the human placenta resemble those of skeletal muscle, both in endothelial cell junctional organization and in the single passage extraction of radiolabeled tracers. Addition of histamine (100 microM) to the fetal perfusate in an isolated term lobule resulted in a rapid and sustained rise (40-80%) over a 30-min perfusion period in the single circulation extraction values of 57Co-labeled cyanocobalamin and of 51Cr-labeled EDTA, but not of 22Na. Extraction values for 125I-albumin were not increased in histamine-perfused vessels nor was any focal leakage of label observed in serial cryostat sections of lobules perfused with rhodamine-conjugated albumin. There was no electron microscopic evidence of transendothelial channels in the microvascular bed; no gaps were seen at the paracellular cleft regions or in neighboring cytoplasm in any microvessels. Tilting of sections on a goniometric stage showed a significant increase in the separation between adjoining endothelial membrane leaflets at tight junctional regions (from 4.1 to 6.1 nm) although the dimensions of the wide zones remained unchanged. Placental microvessels contain the endothelial adhesion molecules PECAM-1 and VE-cadherin in the wide regions of paracellular clefts: PECAM-1 is also localized on the luminal membrane. Histamine-stimulated microvessels showed an altered staining pattern for both of these molecules, at the expense of the cleft regions. These changes in adhesion molecule distribution and tight junctional membrane separation may be parts of a series of events which leads to increased permeability during inflammatory events.

Antigens, CD↗
PubMed1991

The role of inflammation in nonspecific abdominal aortic aneurysm disease.

The predominant pathologic feature of abdominal aortic aneurysm is elastin destruction, and elastin destruction may be mediated by inflammation. In this investigation serial sections of abdominal aortic aneurysm specimens were selectively stained to study the relationship between inflammation and elastin degradation. In addition, soluble aortic extracts were examined for the presence of immunoglobulins. An inflammatory infiltrate was present in 8 of 10 of the abdominal aortic aneurysm specimens examined. The infiltrate was mononuclear, commonly located at the junction of the media and adventitia; it did not codistribute with loss of elastin. The presence of an inflammatory component in abdominal aortic aneurysm was associated with a large amount of immunoglobulin in soluble extracts from aneurysmal tissue compared to atherosclerotic and normal control extracts. This study further characterizes the microscopic pathology of abdominal aortic aneurysm and describes the presence of immunoglobulin in soluble tissue extracts. In addition, the possible role of inflammation in abdominal aortic aneurysm as it relates to protease expression is detailed.

Aorta, Abdominal↗
PubMed1988

Diazepam binding inhibitor-like immunoreactivity (DBI-LI) in human CSF. Correlations with neurological disorders.

Cerebrospinal fluid (CSF) levels of the anxiogenic neuropeptide diazepam binding inhibitor (DBI) were determined by radioimmunoassay in 281 patients who underwent evaluation for neurological problems. Serial dilution curves and reverse-phase high pressure liquid chromatography showed that the immunoreactive material in CSF behaved just as authentic DBI extracted from human brain. Furthermore in the assay there was no evidence of interference from CSF samples deprived of DBI by immunoaffinity. In 82 patients with no evidence of major lesions in the central nervous system, who acted as controls, the CSF DBI content was shown to be age- and sex-related. No correlation was observed with the CSF protein concentration. In patients with different types of dementia, the levels of CSF DBI were significantly increased in a group with normal pressure hydrocephalus. No significant differences were found between Alzheimer's disease, multi-infarct dementia, or dementia with Parkinson's disease and controls. In non-demented patients with Parkinson's disease the levels of DBI were increased in a subgroup with depressive disturbances whereas no differences was observed in the non-depressed cases. The content of DBI was markedly reduced in 5 cases with olivopontocerebellar atrophy and in 4 with spinocerebellar ataxia. In all the other disorders studied the levels of DBI were similar to or slightly lower (multiple sclerosis) than those of the controls. The origin of DBI in cerebrospinal fluid is uncertain; a number of various possibilities are discussed concerning the proposed role of DBI as modulator of brain GABAergic transmission.

Adolescent↗
PubMed1975

Bronchoprovocation: effect on priming and desensitization phenomenon in the lung.

Priming, or increased sensitivity to antigen, has not been demonstrated in the lung and could play a role in asthmatic symptomatology during seasonal pollen exposure. It is also an important consideration in the design of any experimental protocol requiring serial bronchoprovocations with antigen. Thirteen patients with a history of asthma symptoms during the pollen season and a positive skin test to ragweed extract were selected. Patients were given bronchial challenge out of season on 4 successive days with stepwise inhalations of antigen, and airways conductance was monitored in the body plethysmograph. Antigen dose-response curves were drawn, and the cumulative dose required for a 35% reduction in specific airway conductance was calculated and designated Provocation Dose (PD35). No regular trend toward either priming or desensitization was noted. The daily changes in antigen sensitivity did not correlate with daily variation of baseline pulmonary function. To determine if there was any priming due to natural exposure to pollen, 9 patients were brought back and rechallenged during the pollen season with no significant increase in bronchial sensitivity to ragweed extract. The PD35 method provides figures useful for comparing dose-response curves and shows a one-log variation from day to day. Any evaluation by bronchial challenge of antigen sensitivity or drug efficacy must take into account such variation.

Aerosols↗
PubMed1989

Transcutaneous oxygen tension measurements during hemorrhagic hypoperfusion using Trendelenburg and the pneumatic antishock garment.

Transcutaneous oxygen tension (PtCO2) was observed during hemorrhagic hypoperfusion using four therapeutic modalities: pneumatic antishock garment (PASG), 20 degrees Trendelenburg positioning, combined PASG-Trendelenburg, and whole blood infusion. Anesthetized mongrel dogs were mechanically ventilated. A heated transcutaneous oxygen sensor was applied to the skin overlying the sternum. Animals were bled over 10 min of 25% of their calculated blood volume. A therapeutic intervention was applied at the onset of hemorrhage (PASG, Trendelenburg, PASG-Trendelenburg or control). All animals were observed for 20 min, then during a 10-min shed blood reinfusion period, and for 20 min thereafter. PtCO2 was measured continuously and the following were measured serially: cardiac output, mean arterial pressure (MAP), mixed venous oxygen tension (MvO2), and arterial oxygen tension (PaO2). Cardiac index (CI) and the oxygen extraction ratio were calculated. PtCO2 decreased immediately after hemorrhage in all animals. Control values remained consistently below values for active interventions during this time. All groups regained baseline levels of PtCO2 after reinfusion of shed blood volume. PaO2 remained nearly constant during all experiments. MAP and CI fell in all groups following hemorrhage but did so less precipitously in the PASG group. The PASG and PASG-Trendelenburg groups showed the greatest increase in CI during reinfusion. These results suggest that when PtCO2 is relied upon as an indicator of adequacy of resuscitation during moderate hemorrhagic shock, that cutaneous perfusion may be improved by the PASG or Trendelenburg position, and that perfusion is most effectively restored by blood infusion.

Animals↗
PubMed2003

Reconstruction of serially acquired slices using physics-based modeling.

This paper presents an accurate, computationally efficient, fast, and fully automated algorithm for the alignment of two-dimensional (2-D) serially acquired sections forming a 3-D volume. The approach relies on the determination of interslice correspondences. The features used for correspondence are extracted by a 2-D physics-based deformable model parameterizing the object shape. Correspondence affinities and global constrains render the method efficient and reliable. The method accounts for one of the major shortcomings of 2-D slices alignment of a 3-D volume, namely variable and nonuniform thickness of the slices. Moreover, no particular alignment direction is privileged, avoiding global offsets, biases, and error propagation. The method was evaluated on real images and the experimental results demonstrated its accuracy, as reconstruction errors were smaller than 1 degree in rotation and smaller than 1 pixel in translation.

Algorithms↗
PubMed2000

Identification and characterization of humic substances-degrading bacterial isolates from an estuarine environment.

Bacterial isolates were obtained from enrichment cultures containing humic substances extracted from estuarine water using an XAD-8 resin. Eighteen isolates were chosen for phylogenetic and physiological characterization based on numerical importance in serial dilutions of the enrichment culture and unique colony morphology. Partial sequences of the 16S rRNA genes indicated that six of the isolates were associated with the alpha subclass of Proteobacteria, three with the gamma-Proteobacteria, and nine with the Gram-positive bacteria. Ten isolates degraded at least one (and up to six) selected aromatic single-ring compounds. Six isolates showed ability to degrade [(14)C]humic substances derived from the dominant salt marsh grass in the estuary from which they were isolated (Spartina alterniflora), mineralizing 0.4-1.1% of the humic substances over 4 weeks. A mixture of all 18 isolates did not degrade humic substances significantly faster than any of the individual strains, however, and no isolate degraded humic substances to the same extent as the natural marine bacterial community (3.0%). Similar studies with a radiolabeled synthetic lignin ([beta-(14)C]dehydropolymerisate) showed measurable levels of degradation by all 18 bacteria (3.0-8.8% in 4 weeks), but mineralization levels were again lower than that observed for the natural marine bacterial community (28.2%). Metabolic capabilities of the 18 isolates were highly variable and generally did not map to phylogenetic affiliation.

Journal Article↗
PubMed1998

The influence of prostaglandins on steroid conversions by human gingival fibroblasts.

The aim of this investigation was to study the effects of prostaglandins E1 and E2 (PGE1 and PGE2) on the accumulation, release and metabolism of C19 steroids by human gingival fibroblasts (HGF) and gingivae, due to their anabolic potential in inflammatory repair. For the accumulation studies, HGF were incubated with 14C-testosterone at timed intervals and the cell-digests analysed for label uptake. The release of 5 alpha-dihydrotestosterone (DHT) by HGF was studied by preincubating the cells with 14C-DHT and reincubating with cold steroid to quantify its release at timed intervals. For the metabolic studies, HGF/gingival tissue were incubated with 14C-testosterone and serial concentrations of PGE1 and PGE2 to study their effects on the synthesis of DHT. The incubations were terminated at 24 h and extracted metabolites were analysed and quantified. The accumulation of 14C-testosterone by human gingival fibroblasts was elevated 3-fold at 24 h by PGE1 (n = 3, p < 0.001; 1-way ANOVA). The release of 14C-DHT was enhanced nearly 2-fold by PGE1 (n = 3, p < 0.001), compared with controls. Both PGE1 and PGE2 caused 2-fold increases in DHT synthesis by HGF and 3-fold increases in 4-androstenedione formation (n = 4, p < 0.001). With the tissue incubations PGE1/PGE2 caused 3-4 fold increases in DHT synthesis (n = 5, p < 0.005; Wilcoxon signed rank statistic for paired observations). Direct stimulation of the accumulation/release and metabolism of these steroids by prostaglandins in gingivae may contribute to the anabolic potential of androgens in inflammatory periodontal disease.

Acute-Phase Reaction↗
PubMed1975

Extraction of sleep-promoting factor S from cerebrospinal fluid and from brains of sleep-deprived animals.

Sleep-promoting factor (factor S) was extracted, partially purified, and concentrated from cerebrospinal fluid and from acid-acetone extracts of brain stem anc cortex of sleep-deprived goats and sheep. 2. Solutes greater than 500 daltons were largely removed by serial ultrafiltrations through molecular sieves (Amicon membranes UM10 and UM05); solutes less than 350 daltons were largely eliminated by gel filtration through Sephadex G10 columns. Sleep-promoting activity was found in a fraction eluted prior to [14C] sucrose marker. 3. Concentrated fraction were infused intraventricularly in rats (0.1 ml in 30 min just prior to 12-h dark cycle) and in rabbits (0.3 ml in 90 min in morning). Sleep-promoting activity was assayed by decrease in nocturnal locomotor activity of rats and by duration and amplitude of slow-wave cortical EEG in rabbits.

Animals↗
PubMed1982

Evaluation of the intradermal allergy test in normal dogs.

In 45 normal dogs, serial dilutions of different allergens were injected intradermally to determine the skin threshold concentrations. For housedust, housedust mite (Dermatophagoides pteronyssinus) and human dandruff extracts these were found to be the same in the dog as in man, while those for animal dandruffs, pollens and fungi were 10 times higher in the dog. The term 'histamine reactive value' (RH) is introduced for the dog. The mean (+/-SD) was found to be 49.0 +/- 2.5 in normal dogs, using histamine diphosphate solutions of 0.01, 0.1 and 1 per cent.

Allergens↗
PubMed1995

Simple quantification of benzodiazepine receptor binding and ligand transport using iodine-123-iomazenil and two SPECT scans.

UNLABELLED: The feasibility of simplified procedures for the quantification of benzodiazepine receptor binding using [123I]iomazenil and SPECT was assessed. METHODS: Six normal male volunteers were studied. Following intravenous injection of 111 MBq [123I]iomazenil, serial dynamic SPECT scanning was performed for 120 min and the concentration of lipophilic compounds in the arterial plasma was determined by chloroform extraction. Kinetic parameters were estimated by a curve-fitting procedure using the following four models: three-compartment, four-parameter (K1-k4) (3C4P), three-compartment, three-parameter (fixed K1/k2) (3C3P), three-compartment, two-parameter (fixed K1/k2 and k4) (3C2P) and two-compartment, two-parameter (K1 and k2) (2C2P). Kinetic parameters were also calculated by a table look-up procedure with 3C2P using only two SPECT data acquisitions. Parametric images were generated based on the same procedure. RESULTS: In all models, the curve-fitting procedure gave similar outcomes for ligand transport (K1) and receptor density parameters (i.e., binding potential or distribution volume). The 3C4P parameters showed significant correlation between k2 and k3, while 3C3P did not show such a correlation, suggesting the stability of 3C3P. The 3C2P provided parameters essentially identical to those with the 3C3P, indicating the suitability of this model, while 2C2P gave similar distribution volume but obviously low K1. CONCLUSION: Table look-up procedures based on the 3C2P model permit quantification of benzodiazepine receptor binding using [123I]iomazenil with two SPECT scans. This method may be of clinical value in the diagnosis of various diseases.

Adult↗
PubMed1996

Further elucidation of pharmacokinetic interaction between diltiazem and warfarin.

Diltiazem and warfarin are both highly protein-bound providing the opportunity for a protein-binding displacement interaction to occur. Additionally, diltiazem has demonstrated the ability to inhibit the metabolic clearance of hepatically cleared drugs (e.g. warfarin). We evaluated these pharmacokinetic mechanisms for a potential interaction. A single warfarin 25 mg oral dose was given with and without oral diltiazem 30 mg 3 times daily to 20 healthy male volunteers in a randomized crossover design with a washout period of at least 6 days. Serial blood samples for INRs and free and total warfarin concentrations were obtained over 144 hours after each warfarin dose. Free fraction was extracted by equilibrium dialysis and free and total warfarin concentrations were analyzed by HPLC. Anticoagulation was assessed using AUC0-6,INR (days). Diltiazem was found not to potentiate anticoagulation by warfarin. Furthermore, warfarin protein binding displacement did not occur. Diltiazem inhibited both free and total warfarin clearance, though not statistically significant. Although slightly inhibiting metabolism of warfarin, based on warfarin clearance, diltiazem did not increase warfarin's anticoagulant effect making it an acceptable addition to therapy if required.

Adult↗
PubMed1996

Coronary flow and flow reserve by PET simplified for clinical applications using rubidium-82 or nitrogen-13-ammonia.

UNLABELLED: To validate routine, noninvasive determination of absolute myocardial perfusion and coronary flow reserve (CFR), cardiac PET was performed in animals using a simplified imaging protocol, high-dose dipyridamole and a simplified quantitative algorithm specific for 82Rb and 13N-ammonia. METHODS: One hundred thirty-five PET scans were obtained in eight dogs after intravenous 13N-ammonia or 82Rb using serial dynamic PET or a simple two-image dataset. A simple flow model using the two-image dataset was developed for each radionuclide to account for varying arterial input function, flow-dependent myocardial extraction and increased permeability surface area (PS) product due to capillary recruitment at high flows not incorporated into previous models. Myocardial perfusion by the simple model was compared to standard, complete, two-compartment kinetic models validated by comparison to electromagnetic flow meter. RESULTS: For 13N-ammonia, myocardial perfusion by the simple PET model correlated with that by complete compartmental analysis of multiple serial PET images with r = 0.94, slope = 0.96; CFR by compartmental analysis correlated with CFR by electromagnetic flow meter with r = 0.94, slope = 0.97. For 82Rb, myocardial perfusion determined by the simple model correlated with that determined by complete compartmental analysis of multiple serial PET images with r = 0.98, slope = 1.06; CFR determined by compartmental analysis correlated with CFR by electromagnetic flow meter with r = 0.88, slope = 1.13. CONCLUSION: A simplified PET protocol using 13N-ammonia or 82Rb and simple flow models provide noninvasive measurement of CFR up to six times baseline flow throughout the heart and diagnostic image quality for routine clinical application.

Ammonia↗
PubMed2006

[Tutorial session using cross sections of upper and lower limbs in human gross anatomy course].

We performed tutorial education in gross anatomy using thick serial cross sections (3-4 cm) of one of the upper or lower limbs of a cadaver donated for use in the macroscopic dissection course. Group learning (four people) proceeded with the extraction of the point in question from an arbitrary section. Consequently students obtained a deeper understanding of the cross sectional structure of an upper or lower limb and further deepened their understanding of the three-dimensional structure of limbs. Each group collected clinical images of the part corresponding to the selected section to present, thus bringing together the relationship between the anatomical knowledge studied from the specimen and the related disease. A questionnaire survey after the practicum suggested that the practice of using serial cross sections and the presentation of relevant images using Microsoft PowerPoint were effective means of the study. Useful points on the student's understanding were collected with respect to the following three opinions: Fresh aspects, Grasp of the position, and Three-dimensional views. Moreover, the dissection program in gross anatomy combined with thick cross sections was thought to be helpful in that not only was observing the cross structure but also to dissect it if necessary, with the students obtaining a deeper understanding of the structure as a result. In our anatomy practice thereafter, the practicum was done with awareness of the issues faced by the students in mind. It was concluded that the practicum that uses the cross sections is an effective means in gross anatomy education.

Anatomy↗
PubMed1988

Natriuretic activity of digoxin-like immunoreactive substance extracted from cord blood.

It was examined whether the digoxin-like immunoreactive substance (DLIS) extracted from cord blood has a natriuretic activity. The DLIS was prepared from cord blood of healthy fullterm infants by acetone-HCl extraction and a gel filtration column. A solution (solution A) containing 1.0 ng/ml of DLIS or another solution (solution B) consisting of solution A from which the DLIS had been completely absorbed by rat brain synaptosome, a crude digoxin receptor, were infused directly into the renal arteries of rats. Serum and urine were serially sampled. The excretion of sodium into the urine increased gradually after the initiation of infusion and reached a level two or three times higher than that before infusion (p less than 0.05). The infusion of a buffer solution or of the extract from which the DLIS had been absorbed by rat brain synaptosome did not significantly increase the urinary excretion of sodium. Statistical analysis showed a clear difference in the natriuretic activity between solutions A and B (p less than 0.01, p less than 0.05). Well-known natriuretic substances such as atrial natriuretic hormone, prostaglandin E2, F2 alpha, bradykinin and oxytocin dopamine were not detected enough to contribute to natriuresis in the extracts. From this data, we speculated that the DLIS in cord blood has a natriuretic activity and that it plays a role in water and sodium homeostasis in perinatal life.

Animals↗
PubMed2005

Evaluation of COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen HBV DNA, HCV RNA and HIV-1 RNA amplification and detection.

BACKGROUND AND OBJECTIVES: This report describes the evaluation of the COBAS AmpliPrep instrument for fully automated generic nucleic acid extraction in conjunction with hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA, and human immunodeficiency virus (HIV)-1 RNA COBAS AmpliScreen amplification and detection using serial dilutions of the WHO international standards (IS) and the PeliCheck reference panels. MATERIALS AND METHODS: Serial diluted samples of the WHO IS and the PeliCheck reference panels were tested 24 times to determine the HBV DNA, HCV RNA and HIV-1 RNA detection limits by Probit analysis. The existence and extent of cross-contamination were assessed by testing alternating high titre HBV DNA-positive and -negative samples. The specificity of the AmpliPrep-AmpliScreen test for HBV was determined by testing 232 minipools consisting of six donations, all negative for HCV/HIV-1 nucleic acid testing (NAT) and HBsAg. In addition, a HBV genotypes A-G panel was tested. RESULTS: The respective 95% detection limits (and 95% CI) on the WHO IS and on the PeliCheck reference panels were 6.7 (4.3-13) IU/ml and 123 (68-301) gEq/ml for HBV DNA, 23 (11-106) IU/ml and 126 (84-233) gEq/ml for HCV RNA, and 187 (108-422) IU/ml and 183 (108-434) gEq/ml for HIV-1 RNA. Based on the WHO IS and the PeliCheck reference panels, no significant differences in sensitivity for HBV and HCV were found between AmpliPrep and the licensed MultiPrep extraction method. The sensitivity of AmpliPrep-AmpliScreen for HIV-1 was probably twofold lower as compared to the MultiPrep-AmpliScreen method. No cross contamination was observed. All 232 minipools were HBV NAT-negative. The AmpliPrep-AmpliScreen test for HBV detected HBV genotypes A-G with equal sensitivity. CONCLUSIONS: The AmpliPrep instrument combined with the AmpliScreen assays for HBV, HCV and HIV-1 is robust and suitable for NAT donor screening. The sensitivity criteria for HIV-1 and HCV as defined by the Paul Ehrlich Institute and the Food and Drug Administration for minipool NAT screening are met by this system. SINGLE SENTENCE SUMMARY: Generic COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen detection for HBV, HCV and HIV-1.

DNA, Viral↗