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Biomedical subjects

M R Gray

Publications and source records attributed to M R Gray.

68 records · Page 4Linked to original sources
PubMed1987

High-performance liquid chromatography of lidocaine and nine of its metabolites in human plasma and urine.

A simple high-performance liquid chromatographic procedure for quantifying lidocaine and nine of its metabolites in human plasma and urine is described. This method involves aqueous acetylation of the metabolites with acetic anhydride under mild basic conditions (pH approximately 8.5). Lidocaine and the derivatized metabolites are extracted under basic conditions with ethyl acetate. Calibration curves constructed for these compounds are linear in the concentration range studied (0.2-500 micrograms/ml). The relevance of this method is shown by measuring the plasma and urine profiles of two patients with suspected myocardial infarction.

Acetylation↗
PubMed1985

Rapid serodiagnosis of leptospirosis using the IgM-specific Dot-ELISA: comparison with the microscopic agglutination test.

The dot enzyme-linked immunosorbent assay (Dot-ELISA) was compared to the microscopic agglutination test (MA test) for the diagnosis of human leptospirosis. Of 177 sera from 68 soldiers who trained in the Republic of Panama, 102 sera were positive in the MA test and 93 of these sera were positive in the IgM-specific Dot-ELISA. Incidence of infection was 50 of 68 patients with the MA test and 48 of 68 in the IgM Dot-ELISA. Five MA test-positive sera were reactive only in the IgG-specific Dot-ELISA, suggesting previous exposure. All 21 infecting serovars of Leptospira interrogans, as determined by positive reactions in the MA test or culture of blood and urine specimens, were reactive in the Dot-ELISA. Of 75 sera negative in the MA test, 61 were nonreactive in the Dot-ELISA. However, 9 of these 14 Dot-ELISA-positive/MA test-negative sera were acute samples from patients whose later sera were MA test-positive. Positive reactions in the IgM Dot-ELISA occurred in 2 of 30 control, 4 of 10 Lyme disease, 1 of 11 relapsing fever, and 1 of 8 yaws sera; 10 syphilis patient sera were nonreactive. The IgM-specific Dot-ELISA appears to be sensitive and specific for the serodiagnosis of acute leptospirosis. In addition, this rapid test is inexpensive, simple to perform, utilizes minute volumes of killed leptospiral antigen and is easily adaptable to field use.

Agglutination Tests↗
PubMed1984

An efficacy trial of doxycycline chemoprophylaxis against leptospirosis.

Because leptospirosis has been an important cause of morbidity in U.S. soldiers training in the Republic of Panama, we conducted a randomized, double-blind, placebo-controlled field trial during the fall of 1982 to determine whether doxycycline was an effective chemoprophylactic agent against this infection. Doxycycline (200 mg) or placebo was administered orally on a weekly basis and at the completion of training to 940 volunteers from two U.S. Army units deployed in Panama for approximately three weeks of jungle training. Twenty cases of leptospirosis occurred in the placebo group (an attack rate of 4.2 per cent), as compared with only one case in the doxycycline group (attack rate, 0.2 per cent, P less than 0.001), yielding an efficacy of 95.0 per cent. This study demonstrated the value of doxycycline as a prophylactic drug against leptospirosis.

Clinical Trials as Topic↗
PubMed1984

Drosophila maternal and embryo mRNAs transcribed from a single transcription unit use alternate combinations of exons.

We have investigated the organization and transcription of several genes in Drosophila melanogaster which are clustered on an 18-kb cloned DNA fragment (c25) that maps at 99D on the cytogenetic map. Multiple mRNAs, transcribed from genes which lie adjacent to a ribosomal protein ( rp49 ) gene, are present during oogenesis, embryogenesis, or both. At least five mRNAs are transcribed from one of these genes ( EH8 ); three are zygotic transcripts, of which two are blastoderm stage-specific, whereas two others accumulate during oogenesis and are therefore matenal mRNAs. The complex transcription pattern of this gene indicates that alternate usage of protein-coding exons results in the production of different mRNAs with different coding capabilities during oogenesis and embryogenesis. The EH8 transcription unit is framed by genes actively expressed in the adult male fly; thus, the blastoderm stage-specific promoter may be silent although within a region of transcriptionally active chromatin.

Amino Acid Sequence↗
PubMed1982

Open reading frame cloning: identification, cloning, and expression of open reading frame DNA.

A plasmid was constructed that facilitates the cloning and expression of open reading frame DNA. A DNA fragment containing a bacterial promoter and the amino terminus of the cI gene of bacteriophage lambda was fused to an amino-terminally deleted version of the lacZ gene. An appropriate cloning site was inserted between these two fragments such that a frameshift mutation was introduced upstream of the lacZ-encoding DNA. This cloning vehicle produces a relatively low level of beta-galactosidase activity when introduced into Escherichia coli. The insertion of foreign DNA at the cloning site can reverse the frameshift mutation and generate plasmids that produce a relatively high level of beta-galactosidase activity. A large fraction of these plasmids produce a fusion protein that has a portion of the lambda cI protein at the amino terminus, the foreign protein segment in the middle, and the lacZ polypeptide at the carboxyl terminus. The production of a high level of beta-galactosidase and a large fusion polypeptide guarantees the cloning of a DNA fragment with at least one open reading frame that traverses the entirety of the fragment. Hence, the method can identify, clone, and express (as part of a larger fusion polypeptide) open reading frame DNA from among a large collection of DNA fragments.

Base Sequence↗
PubMedDate not supplied

Effect of dissolved oxygen control on growth and antibiotic production in Streptomyces clavuligerus fermentations.

A proportional-integral control system was used to control dissolved oxygen in a fermentor at constant shear and mass transfer conditions. Growth and antibiotic production in Streptomyces clavuligerus were studied at different dissolved oxygen levels during the fermentation. Three protocols were employed: no-oxygen control to provide a base case, oxygen controlled to a preset saturation level throughout the fermentation, and oxygen controlled at a high level only during the growth phase. The last protocol was aimed at optimizing the consumption of oxygen. Lower specific growth rates and cephamycin C yields were obtained when dissolved oxygen was controlled at 50% throughout the fermentation, compared to the base case. A 2.4-fold increase in the final cephamycin yield was observed when dissolved oxygen was controlled at saturation levels during the growth phase, compared to the experiments without dissolved oxygen control. This enhancement in yield was independent of the dissolved oxygen (DO) level after exponential growth, in the range of 50-100% saturation. The most effective control strategy, therefore, was to control DO only during active growth when the biosynthetic enzymes were probably synthesized.

Anti-Bacterial Agents↗
PubMedDate not supplied

Mechanisms of lidocaine kinetics in the isolated perfused rat liver. II. Kinetics of steady state elimination.

The steady state kinetics of lidocaine and its metabolites were modeled using nonlinear elimination pathways for multiple enzymes. The main metabolites, monoethylglycinexylidide and 3-hydroxy-lidocaine, were infused in the absence of lidocaine to measure the kinetic parameters for secondary elimination. Data from continuous perfusion of lidocaine in the isolated perfused rat liver at concentrations ranging from 9.6 to 278 microM (N = 16) were used to calculate the kinetic parameters for formation of the main metabolites. The elimination of lidocaine in the liver was approximated by the well stirred model. The whole liver study gave higher elimination rates than were predicted from microsomal studies. The major pathways for elimination of lidocaine in the rat were deethylation and hydroxylation, and subsequent elimination along these pathways accounted for the poor material balance at low dosage levels. The observed competitive inhibition of hydroxylation was in agreement with the predictions of the model.

Animals↗
PubMedDate not supplied

Mechanisms of lidocaine kinetics in the isolated perfused rat liver. III. Evaluation of liver models for time-dependent behavior.

Three models for the elimination of lidocaine in the isolated perfused rat liver were used to simulate the time course of both lidocaine and its metabolites in a single-pass perfusion system: well stirred, parallel tube, and a two-compartment model to test the effects of enzyme heterogeneity. All models included multiple enzymes and multiple metabolic pathways, as well as varying degrees of tissue binding. Although the well stirred and parallel tube models gave qualitatively different results, neither model predicted that the concentration of monoethylglycinexylidide would pass through a maximum and then decline to a lower steady state value, as observed in continuous perfusion experiments. Although each of the three models tested would give reasonable agreement with steady state observations, the test of the time-dependent behavior of both lidocaine and monoethylglycinexylidide was more discriminating. Each model gave characteristic predictions for the time course of the metabolites.

Animals↗
PubMedDate not supplied

The series-compartment model for hepatic elimination.

An empirical model is presented for hepatic elimination of substrates which treats the liver as a series of compartments. The number of compartments serves as an adjustable parameter and determines the effect of changes in blood flow on elimination. By allowing variable dependence of clearance on blood flow, the model serves as a bridge between the extreme cases of venous equilibrium (the "well-stirred" model) and the "parallel-tube" model. The adjustable parameter in the model can be determined by measuring the clearance at different rates of blood flow, or at different extents of binding of the substrate to blood components. Examples are given for steady state elimination of several different substrates with linear and nonlinear elimination kinetics.

Binding Sites↗
PubMedDate not supplied

Kinetic interactions of lidocaine, diphenhydramine, and verapamil with diltiazem. A study using isolated perfused rat liver.

The effect of several tertiary amines, which are known enzyme inhibitors, on the disposition of diltiazem (DZ) was evaluated using a single-pass isolated rat liver perfusion system. Coinfusion of lidocaine (LID) or diphenhydramine (DPH) at the steady state of DZ resulted in a sharp increase in the perfusate concentration of DZ, which was followed by a decline to a new steady-state concentration (Cnss) that was higher than the original Cnss value (46 and 45%, respectively). The initial sharp increase in DZ concentration was attributed to the displacement of DZ from its tissue binding sites; the higher Css values were due to the inhibition of N-oxidation and O-demethylation, and some unknown primary metabolic pathways. The kinetics of LID were altered by DZ; the steady-state extraction ratio of LID was reduced and the characteristic maximum in the concentration-time profile of its N-deethylated metabolite, MEGX, was abolished. These results suggest that DZ and LID share common isozymes in their disposition and that the two drugs are also capable of inactivating similar enzymes. The effect of enzyme inactivation on DZ disposition was evaluated by intraperitoneal pretreatment of rats with either saline (0.4 ml) or 1 of the 4 drugs--DZ (20 mg/kg), LID (30 mg/kg), DPH (20 mg/kg), and verapamil (10 mg/kg)--daily for at least 3 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Amines↗
PubMedDate not supplied

Mechanisms of time-dependent kinetics of diltiazem in the isolated perfused rat liver.

The time-dependent mechanisms of diltiazem (DZ) disposition were studied in a single-pass isolated perfused rat liver system. DZ (2-100 microM) was infused continuously until a steady state was achieved. The time required to achieve steady state (Tss) ranged from 15 to 75 min and was inversely related to infusion concentration. Steady-state extraction (E) of DZ decreased from 0.98 to 0.73 as the outlet concentration increased from 0.03 to 26.34 microM. This reduction of E is not related to the saturation of the primary deacetylation and N-demethylation pathways of metabolism. The N-demethylated metabolite went through a characteristic maximum prior to reaching a steady state, indicating enzyme inactivation. During a second DZ infusion, spaced with a 30-min washout period (stop-infusion experiment), the concentration vs. time profile of DZ was similar to that of the first one. Washout data from stop-infusion and [3H]DZ infusion studies showed that DZ and its metabolites were tightly bound to liver proteins. This observation is consistent with the unusually long Tss of DZ. Infusion studies with [3H]DZ showed that 669.5 +/- 156.5 and 974.2 +/- 99.2 nmol of DZ and its metabolites (calculated as DZ) were bound and/or distributed/g of liver at inlet concentrations of 35.5 +/- 3.2 and 67.2 +/- 3.4 microM, respectively. The amounts of DZ and its metabolites bound irreversibly to the whole liver, hepatic microsomal and hepatic cytosolic proteins were not different at the two inlet concentrations studied and were 24.5 +/- 6.6, 48.8 +/- 11.8, and 23.7 +/- 5.8 pmol/mg of protein, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗