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Biomedical subjects

J A Blake

Publications and source records attributed to J A Blake.

34 records · Page 2Linked to original sources
PubMed1996

Coexpression of a human P450 (CYP3A4) and P450 reductase generates a highly functional monooxygenase system in Escherichia coli.

The catalytic activities of recombinant cytochrome P450s expressed in E. coli have been impeded by the absence of endogenous P450 reductase. To solve this problem, we coexpressed P450 reductase with CYP3A4. Membranes from this strain contained 215 pmol P450/mg protein and a reductase activity of 1315 nmol cytochrome c reduced/min per mg. We detected 6beta-hydroxylation of testosterone and oxidation of nifedipine in vivo with turnover numbers of 15.2 and 17.3 min(-1), respectively. These values compare favourably with those obtained using an optimally reconstituted system. Our data demonstrate that a catalytically efficient human P450 system can be generated in E. coli.

Base Sequence↗
PubMed1996

Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii.

The complete 1.66-megabase pair genome sequence of an autotrophic archaeon, Methanococcus jannaschii, and its 58- and 16-kilobase pair extrachromosomal elements have been determined by whole-genome random sequencing. A total of 1738 predicted protein-coding genes were identified; however, only a minority of these (38 percent) could be assigned a putative cellular role with high confidence. Although the majority of genes related to energy production, cell division, and metabolism in M. jannaschii are most similar to those found in Bacteria, most of the genes involved in transcription, translation, and replication in M. jannaschii are more similar to those found in Eukaryotes.

Amino Acid Sequence↗
PubMed1995

Prodrugs to improve the oral bioavailability of a diacidic nonpeptide angiotensin II antagonist.

DMP 811 is a diacidic angiotensin II antagonist. It has relatively low oral bioavailability in rats. A prodrug approach to improving oral bioavailability was tested. Five esters were synthesized and their stability in rat plasma in vitro was determined. The hydrolysis rates of these five esters ranged from almost immediate to negligible. A simple n-propyl ester was hydrolyzed very slowly (< 10% in 24 hr) in rat plasma in vitro, and after oral dosing in rats plasma prodrug concentrations were much greater than DMP 811 concentrations. A pivaloyloxymethyl ester (1) was hydrolyzed relatively rapidly in rat plasma in vitro. Prodrug 1 was rapidly hydrolyzed by the intestine in vitro, and the intestinal permeation of DMP 811 was increased. DMP 811 oral bioavailability was 47% in rats dosed with 10 mg/kg 1, compared to 11% for rats dosed with 10 mg/kg DMP 811. However, DMP 811 bioavailability was only 27% after a 2 mg/kg dose of 1. In vitro plasma hydrolysis of 1 was highly species-dependent, with a half-life of 13 hr in human plasma but only 1 min in rat plasma. The prodrug approach has potential for improving the oral bioavailability of DMP 811, but selection of the optimal prodrug must be done in humans or in a species, such as dogs, with hydrolysis characteristics closer to humans.

Administration, Oral↗
PubMed1991

An in vitro evaluation of metabolism and poor membrane permeation impeding intestinal absorption of leucine enkephalin, and methods to increase absorption.

The goals of this study were to evaluate how metabolism and poor membrane permeability act as barriers to absorption of a model peptide, leucine enkephalin (YGGFL), and how those barriers can be overcome. The in vitro everted rat intestine method was used. YGGFL was rapidly metabolized when exposed to the mucosa of the jejunum, and destyrosyl leucine enkephalin was formed. Metabolism was slower for ileal intestinal segments, however, and was almost absent when colonic segments were used. The aminopeptidase inhibitor boroleucine, an aminoboronic acid derivative, reduced the YGGFL metabolism rate 2-fold at 1/100th the concentration of substrate when the intestine was simultaneously exposed to substrate and inhibitor. Pretreatment of the intestine with boroleucine further reduced the metabolism rate. Thiorphan, an inhibitor of endopeptidase 24.11, had additive inhibitory effects with boroleucine. Inhibition of metabolism alone did not enable YGGFL to permeate the membrane. The permeability enhancer, sodium glycocholate, also did not alone enable membrane permeation. Substantial YGGFL permeated the membrane only when metabolism was inhibited and permeability was enhanced. EDTA had both these effects.

Animals↗
PubMed1990

Transdermal oxymorphone formulation development and methods for evaluating flux and lag times for two skin permeation-enhancing vehicles.

Oxymorphone is a candidate for transdermal delivery since it is a very potent analgesic, is not very effective orally, and has a short duration of action. In developing a transdermal delivery system, two criteria that were considered important were achieving adequate flux and minimizing the lag time. Oxymorphone skin permeation rates in vitro were very low unless skin permeation enhancers were included in the vehicle. After an initial screen of 17 formulations, two skin permeation-enhancing formulations were selected for further study. These were myristic acid:propylene glycol:oxymorphone base (A), and decylmethylsulfoxide:ethanol:water:oxymorphone.HCl (B). With either formulation and either human or hairless guinea pig skin, there was little dependence of either in vitro flux or lag time on the section of skin used (stratum corneum, epidermis, epidermis/dermis). There were significant differences between human skin and hairless guinea pig skin when comparing in vitro fluxes with the two formulations. With formulation A, fluxes through hairless guinea pig skin were three-to fivefold greater than through human skin. With B, however, fluxes through human skin were up to fivefold greater than through hairless guinea pig skin. In vitro lag times with A were generally long (approximately 24 h), whereas those with B were much lower (approximately 1 to 10 h). The species dependence of permeation enhancement and the differences in lag time between formulations could be related to differences in the mechanisms of permeation enhancement. In vivo lag times with the fatty acid:propylene glycol vehicle were estimated in hairless guinea pigs based on plasma oxymorphone concentrations. These were much lower than in vitro lag times.

Administration, Cutaneous↗
PubMed1990

Contributions of drug solubilization, partitioning, barrier disruption, and solvent permeation to the enhancement of skin permeation of various compounds with fatty acids and amines.

The contributions of several proposed mechanisms by which fatty acids and amines might increase skin permeation rates were assessed. Permeation rates of model diffusants with diverse physicochemical properties (naloxone, testosterone, benzoic acid, indomethacin, fluorouracil, and methotrexate) through human skin were measured in vitro. The enhancers evaluated were capric acid, lauric acid, neodecanoic acid, and dodecylamine. Increased drug solubility in the vehicle, propylene glycol (PG), in some cases accounted for the increases in flux in the presence of adjuvants, since permeability coefficients were unchanged. Partition coefficients of some drugs into isopropyl myristate or toluene were increased by the adjuvants, but this did not occur for combinations of an acid with a base (adjuvant-drug or drug-adjuvant). Increases in flux not accounted for by increases in drug solubility or partitioning were assumed to involve disruption of the barrier function of skin (increased skin diffusivity). All fatty acids increased skin diffusivity of naloxone, testosterone, indomethacin, and fluorouracil but not of methotrexate or benzoic acid. Dodecylamine increased skin diffusivity only for fluorouracil. Capric acid and dodecylamine, but not lauric acid or neodecanoic acid, increased the skin permeation rate of PG, suggesting that enhanced solvent penetration could also be involved as a mechanism for increased skin permeation of the drug. However, the increase in PG flux due to dodecylamine was nullified when methotrexate was added to the vehicle, possibly because of a dodecylamine/methotrexate interaction. These studies demonstrate that drug solubilization in the vehicle, increased partitioning, increased solvent penetration, and barrier disruption each can contribute to increased skin permeation rates in the presence of fatty acids and amines.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Pharmaceutic↗
PubMed1990

Effects of plasma protein binding displacement on the pharmacokinetics, tissue and tumor concentrations and efficacy of brequinar, a highly protein-bound antitumor agent.

Brequinar is a developmental antitumor agent which is highly bound to plasma proteins. The effects of plasma protein binding displacement on brequinar pharmacokinetics, tissue distribution, tumor distribution and antitumor efficacy were evaluated. Sodium salicylate and ibuprofen increased the percentage of free brequinar in serum in vitro, in proportion to their added concentrations. Sodium salicylate also altered the pharmacokinetics of i.v. brequinar in rats when adminstered i.v. or p.o. at 10- or 50-fold higher doses than brequinar. At the highest salicylate/brequinar dose ratio, significant increases were observed for terminal half-life, mean residence times in the body and tissues, systemic clearance, distribution clearance, the volume of the central compartment and volume of distribution at steady state. Neither salicylate nor ibuprofen increased brequinar concentrations in lung and muscle specimens from rats, 4 or 24 hr after dosing. Tumor, lung and muscle brequinar concentrations in mice were also unaffected by coadministered sodium salicylate, 4 or 24 hr after a single i.v. brequinar dose. In rats infused for 48 hr with brequinar or brequinar plus salicylate, salicylate increased the percentage of free brequinar in plasma and lungs, but total brequinar concentrations were reduced. Antitumor efficacy was evaluated by measuring the survival times of mice implanted with L1210 leukemia cells. Salicylate-treated mice had a similar brequinar dose/response profile as mice not coadministered salicylate. Ibuprofen also did not increase brequinar's antitumor potency.

Animals↗
PubMed1978

Recent progress in dietary fiber (plantix) in human nutrition.

Dietary fiber was extensively discussed in an article by G.A. Spiller and R.J. Amen in this journal in 1975 (Volume 7 Issue 1). The progress in this field has been tremendous in the past 2 years. What was an uncertain field in 1975 and what at that time to some investigators still appeared as a hypothesis or fad without much proof has turned into a much more respected part of nutritional sciences. The scientific quality of recent studies on dietary fiber in human nutrition shows a great deal of sophistication and care. Improved analytical methodologies are being used more extensively. Many nutritionists and clinicians have accepted the fact that a reasonable increase in the dietary fiber intake in the U.S. and U.K would be advisable. In this article, the possible beneficial effects of dietary fiber on health, the possible harmful side effects, and the food science aspects are carefully discussed after an update on chemistry, analytical procedures, and nomenclature. Undoubtedly, dietary fiber has found a new niche in the sciences of nutrition, medicine, epidemiology, and foods.

Adult↗
PubMed1978

Death by hand grenade: altruistic suicide in combat.

Durkheim's theory of suicide is examined as it bears on the concept of altruistic suicide. A subtype called heroic altruistic suicide or self-sacrifice is identified. Relationships are hypothesized between cohesion and self-sacrifice and rank and self-sacrifice, and they are tested in combat situations, utilizing official published information on the Congressional Medal of Honor. Altruistic suicide is found to be higher in more cohesive than in less cohesive groups and more likely among enlisted men than among career officers and noncommissioned officers. In addition to testing hypotheses empirically, this study elaborates Durkheim by extending his analysis of altruistic suicide and clarifies his work by delineating sources of error in his hypotheses.

Adult↗