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Biomedical subjects

R L Smith

Publications and source records attributed to R L Smith.

At least 361 records · Page 20Linked to original sources

Flowthrough reactor flasks for study of microbial metabolism in sediments.

Flowthrough reactor flasks are described that allow continuous low-level nutrient input to mixed anoxic sediments without dilution of the sediment. The flasks were tested by simulating sulfate inputs into sediments collected from a freshwater eutrophic lake. After an initial 2-day adaptation within the reactor system, rates of methane production and sulfate consumption were constant for the duration of a 12-day incubation. A sulfate input rate of 0.15 mmol liter of sediment day resulted in an equivalent rate of sulfate removal, which was unaffected by inputs of acetate (1.0 mmol liter of sediment day). The rate of methane production in control reactors, 0.18 mmol liter of sediment day, was doubled by the addition of acetate, whereas sulfate consumption was only stimulated by additions of high concentrations of sulfate plus acetate (1.5 and 1.0 mmol liter of sediment day, respectively). The reactor system appears to be effective in maintaining the balance between sulfate reduction and methane production in freshwater sediments and is potentially useful for study of the response of sediment populations to varying inputs of naturally occurring substrates, selected inhibitors, or xenobiotic compounds.

Journal Article↗

Alteration of the Fe protein of nitrogenase by oxygen in the cyanobacterium Anabaena sp. strain CA.

Changes in protein composition were noted when heterocysts of Anabaena sp. strain CA were isolated from filaments grown in 1% CO2-99% N2 and subsequently exposed to oxygen. Immunospecific Western blot analysis showed that the Fe protein of nitrogenase is altered. In cells grown under microaerobic conditions, the Fe protein was found in a form with an apparent molecular weight of 30,000. Exposure to oxygen caused a shift in the migration of this polypeptide to a position corresponding to an apparent molecular weight of 31,500. This modification was reversible upon removal of oxygen from the culture. Chloramphenicol did not inhibit the alteration in either direction. Suppression by ammonium nitrate of the recovery of nitrogenase activity from the effects of oxygen did not prevent the alteration of the protein. Other inhibitors of nitrogenase activity, (metronidazole, carbonyl cyanide m-chlorophenylhydrazone, and phenazine methosulfate) were tested for their effect on Fe protein modification. Alteration of the Fe protein may relate to the protection of nitrogenase from the deleterious effects of oxygen.

Cyanobacteria↗

The bioavailability of phenylbutazone in the horse.

[phenyl-14C]-Phenylbutazone was administered to 2 horses p.o. and i.v. on separate occasions. Plasma levels and urinary and faecal elimination of 14C were monitored for up to 7 days after dosing. Phenylbutazone was rapidly and extensively absorbed after oral administration, and its bioavailability was 91% assessed by comparison of plasma AUCs of unchanged drug after p.o. and i.v. administration. The plasma elimination half-life of phenylbutazone was 9.7 h and this was independent of the route of administration. The pattern of elimination of phenylbutazone was independent of the route of administration, with 55% of the dose being found in the urine in 3 days and a further 39% in the faeces in 7 days. These data, which are the first reports of the absolute bioavailability and excretion pathways of phenylbutazone in the horse, are discussed in terms of their significance for the gastrointestinal toxicity of this drug.

Administration, Oral↗

The metabolism of 14C-labelled trimethylamine and its N-oxide in man.

The metabolism and elimination of 14C-labelled trimethylamine and its N-oxide (100 mg orally) were studied in three male volunteers. For both compounds the urine was the major route of elimination, with 95% of the administered 14C being voided in the first 24 h. No radioactivity was found in expired air. The majority (greater than 95%) of the urinary 14C from both compounds was excreted as trimethylamine N-oxide.

Biotransformation↗

Studies on the metabolism of arylacetic acids. 6. Comparative metabolic conjugation of 1- and 2-naphthylacetic acids in the guinea pig, mouse, hamster and gerbil.

The patterns of metabolic conjugation of the isomeric 1- and 2-naphthylacetic acids have been compared in guinea pig, mouse, hamster and gerbil. Equimolar doses of [carboxy-14C]1- and 2-naphthylacetic acids were given to these species by i.p. injection, their urine collected and urinary metabolites examined by t.l.c. before and after treatment with beta-glucuronidase or mild alkali. 2. Urinary excretion of 14C following administration of 14C-1-naphthylacetic acid was 76-93% of dose in 72 h, the bulk being eliminated in 24 h. Urinary metabolites comprised 1-naphthylacetic-glycine and -glucuronide together with the unchanged acid. 3. Following administration of 14C-2-naphthylacetic acid, some 68-94% of the 14C dose was recovered in the urine in 72 h, with the majority in the 0-24 h urine. All four species excreted 2-naphthylacetyl-glucuronide and -glycine: additionally, 2-naphthylacetyl-taurine was excreted by mouse, gerbil and hamster and the glutamine conjugate was also present in hamster urine.

Animals↗

Studies on the metabolism of arylacetic acids. 7. The influence of varying dose size upon the conjugation pattern of 2-naphthylacetic acid in the guinea pig, mouse and hamster.

1. The influence of dose size upon the metabolic conjugation of 2-naphthylacetic acid with various amino acids and glucuronic acid has been studied in the guinea pig, mouse and hamster. 2. Guinea pigs conjugated 2-naphthylacetic acid with glycine and glucuronic acid. 3. Mice conjugated 2-naphthylacetic acid with glycine, taurine and glucuronic acid. Taurine conjugation had the highest capacity, and both this and the glycine mechanism were saturated at doses above 100 mg/kg. 4. Hamsters utilized glutamine, glycine, taurine and glucuronic acid for the conjugation of 2-naphthylacetic acid. No conjugation pathway was saturated by doses up to 200 mg/kg. 5. The thus-far unique ability of 2-naphthylacetic acid to evoke multiple amino acid conjugations, using the taurine and glutamine mechanisms hitherto unknown in these species, appears to be due to its affinity for previously unrecognized enzyme systems, rather than to saturation of 'normal' pathways revealing novel routes at high doses.

Animals↗

The metabolic disposition of [methoxy-14C]-labelled trans-anethole, estragole and p-propylanisole in human volunteers.

1. The metabolic fates of the naturally occurring food flavours trans-anethole and estragole, and their synthetic congener p-propylanisole, have been investigated in human volunteers using the [methoxy-14C]-labelled compounds. The doses used were close to those encountered in the diet, 1 mg, 100 micrograms and 100 micrograms respectively. 2. In each case, the major routes of elimination of 14C were in the urine and in the expired air as 14CO2. 3. Urinary metabolites were separated by solvent extraction, t.l.c. and h.p.l.c., and characterized by comparison of chromatographic mobilities with standards and by radioisotope dilution. Nine 14C urinary metabolites were found after trans-anethole administration, four after p-propylanisole and five after estragole. All were products of side chain oxidations. 4. The principal metabolites of p-propylanisole were 4-methoxyhippuric acid (12%) and 1-(4'-methoxyphenyl)propan-1-ol (2%) and -2-ol (8%). 5. The major metabolite of trans-anethole was 4-methoxyhippuric acid (56% of dose), accompanied by much smaller amounts of the two isomers of 1-(4'-methoxyphenyl)propane-1,2-diol (together 3%). 6. After estragole administration, the two volunteers eliminated 0.2 and 0.4% of the dose respectively as 1'-hydroxyestragole. 7. The human metabolic data is discussed with reference to the comparative metabolic disposition of these compounds in the mouse and rat, species commonly used in their safety assessment.

Adult↗

Metabolizable energy content of pearl millet [Pennisetum americanum (L.) Leeke].

Three experiments were conducted to determine the nitrogen-corrected metabolizable energy (MEn) content of ground pearl millet [Pennisetum americanum (L.) Leeke]. On a dry matter basis, 40% ground pearl millet was substituted for an equivalent amount of glucose monohydrate (reference substance) to determine MEn using male broiler chicks. Food and excreta samples were analyzed for nitrogen, gross energy, chromic oxide, and dry matter. On a dry matter basis, the MEn content of the ground pearl millet ranged from 2.891 to 3.204 kcal/g, depending upon the pearl millet cultivar used. These results suggest that previously reported MEn values for pearl millet may be underestimated by up to 21%.

Animal Feed↗

Spontaneous secretion of a proteoglycan releasing factor by mononuclear cells in juvenile arthritis.

Patients with juvenile arthritis (JA) spontaneously produced a substance which accelerated proteoglycan loss from cultured articular cartilage. Peripheral blood mononuclear cells (PBMC) from 15 patients with JA were cultured for varying days, and cell-free PBMC conditioned media were added to articular cartilage cultures. Release of proteoglycan and collagen from cartilage was quantified by analysis of chondroitin sulfate and hydroxyproline content, respectively, after 4 days of culture. Conditioned media from PBMC of patients with systemic onset JA (3/3) and 5/7 patients with polyarticular JA increased release of proteoglycan when added to cartilage cultures. Mitogen stimulation of the PBMC was unnecessary for activity and addition of mitogen did not alter proteoglycan release. The PBMC conditioned media from the other patients (2/7) with polyarticular JA, from patients (3/3) with systemic onset JA which had progressed to polyarticular JA, and from patients with pauciarticular JA, did not enhance proteoglycan release without mitogen stimulation. PBMC of normal children produced media which enhanced proteoglycan release after mitogen stimulation. No conditioned medium accelerated proteoglycan release if cartilage was freeze killed before culture and none tested reduced cartilage collagen content.

Arthritis↗

The effect of antibiotics on the destruction of cartilage in experimental infectious arthritis.

UNLABELLED: In joints with bacterial arthritis, continuing prolonged destruction of cartilage may occur in spite of prompt, effective antibiotic therapy. We measured the extent to which early antibiotic therapy with ceforanide altered the degradation of the cartilage after arthritis due to Staphylococcus aureus had been produced in the knee joint in rabbits. Degradation of the cartilage was quantified by analyses for glycosaminoglycan and collagen. Three weeks after the infection was produced, the cartilage had lost more than half of its glycosaminoglycan whether the antibiotic therapy had been started at one, two, or seven days after infection. Beginning the antibiotic treatment one day after infection reduced over-all loss of collagen by 37 per cent and decreased the area of erosion of the infected articular surfaces. When antibiotic treatment was begun at four, eight, or twelve hours after infection, the loss of glycosaminoglycan averaged 18 per cent. Prophylaxis with antibiotics completely prevented any degradation of the cartilage. CLINICAL RELEVANCE: The findings reported here show how rapidly cartilage loses glycosaminoglycan when it is involved by arthritis caused by staphylococci and how early treatment of the infection reduces the loss of collagen. There is less protection against loss of glycosaminoglycan. The results emphasize the need for early diagnosis and treatment of infectious synovitis and support the rationale for early administration of antibiotics without waiting for identification of the responsible bacteria.

Animals↗

Two plasma fibronectin fragments with different gelatin-binding properties.

Two subtilisin-generated fragments of human plasma fibronectin exhibit significantly different gelatin affinities. One fragment of 60 kdaltons elutes from gelatin-Sepharose, after intact fibronectin, with pH 5.5, 50 mM citrate, 0.1 M NaCl. A 40 kdalton fragment, which can be derived from the 60 kdalton fragment, remains bound to gelatin under the same conditions. Furthermore, upon urea gradient elution, the 60 kdalton fragment dissociates from gelatin before the smaller fragment. Of the two fragments, the 60 kdalton fragment also demonstrates affinity for heparin. Identical amino-terminal sequences for the two fragments indicate that the heparin-binding activity resides in the difference region at the carboxyl-terminal of the 60 kdalton fragment.

Amino Acid Sequence↗

Bacterial arthritis. A staphylococcal proteoglycan-releasing factor.

We report the effect of staphylococci on induction of proteoglycan release from cartilage in organ culture and document the presence of a staphylococcal proteoglycan-releasing factor in the bacterial growth medium. Staphylococci cocultured with cartilage increased release of proteoglycan 3-4-fold within 48 hours. Viable cartilage was essential for maximum loss of proteoglycan. Killed bacteria had no effect on proteoglycan release. Four Staphylococcus aureus strains yielded growth media preparations active in proteoglycan release. Analysis of the proteoglycan subunits released from cartilage showed only limited degradation, and glycosaminoglycan chain length was not significantly altered.

Animals↗

Biochemistry and antigenicity of osteoarthritic and rheumatoid cartilage.

The purpose of this study was to test whether cartilage serves as the source or repository of antigenic components active in the stimulation of inflammation in rheumatoid arthritis through an analysis of peripheral blood lymphocyte proliferation. Articular cartilage samples were obtained from patients with osteoarthritis, rheumatoid arthritis, and ankylosing spondylitis undergoing joint replacement surgery. Each sample was homogenized and characterized biochemically with respect to the content of proteoglycan, collagen, and immunoglobulin. Proteoglycan content of rheumatoid cartilage was reduced by 71% when compared to osteoarthritic cartilage; the proteoglycan content of ankylosing spondylitis cartilage was reduced by 40% when compared to osteoarthritic cartilage. Immunoglobulins were detectable in all cartilage samples when analyzed by ELISA or end-plate titration. Lymphocyte proliferation, quantified by uptake of 3H-thymidine, was unaltered by addition of cartilage fragments, low (saline) and high salt extracts (2.0 M CaCl2), or cartilage residues. Both autologous and heterologous lymphocytes were tested against the cartilage samples with no difference in reactivity. Purified bovine articular proteoglycans and Type II collagen were also inactive. Although tetanus toxoid and phytohemagglutinin were effective stimulants of proliferation, lymphocytes from arthritis patients were suppressed relative to those of normal individuals. Analysis of arthritic articular cartilage by these techniques failed to demonstrate the presence of antigen(s) stimulating proliferation of peripheral blood lymphocytes.

Adult↗

Polymorphism in drug metabolism--implications for drug toxicity.

Genetic polymorphism of the drug metabolism pathways is commonly encountered both for man and laboratory animal species. It is a major source of variable metabolism and linked events such as response to drugs and toxic substances. In man the cytochrome P-450 isozyme system exhibits considerable polymorphism. Several independent genetic polymorphisms regulating metabolic oxidation at C-, N-, and S-centres have been recently characterised. This phenomenon appears to be a powerful factor in determining biochemical individuality with respect to the oxidative metabolism of drugs and responsiveness to therapeutic agents. Of considerable importance is the recognition of the existence of phenotypes within the individual polymorphism, characterised by an impaired ability to effect metabolic oxidation. Evidence suggests that this factor can determine an increased susceptibility to experience exaggerated pharmacological effects and adverse reactions to several drugs. Laboratory animal species also exhibit polymorphism with respect to several drug metabolic pathways but compared with man, this has been less extensively researched. The study of intra-species differences in metabolism of drugs and toxic substances can be of value: when it occurs it may signal its possible occurrence in man and animal strain models of the human metabolic polymorphisms facilitate the laboratory study of inherited susceptibility to toxicants.

Animals↗

Rapid adaptation of auditory-nerve fibers: fine structure at high stimulus intensities.

Simple phenomenological models of rapid adaptation do not account for details in the response of auditory-nerve fibers to the onset of high-intensity sound stimulus. Simple refractory effects, in combination with higher-order discharge-history effects appear necessary to fully account for the oscillations and plateaus observed in post-stimulus time histograms.

Acoustic Stimulation↗