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Biomedical subjects

C Meredith

Publications and source records attributed to C Meredith.

46 records · Page 3Linked to original sources
PubMed1990

The effect of Biostim (RU-41740) on the expression of cytokine mRNAs in murine peritoneal macrophages in vitro.

The immunomodulatory agent Biostim (RU-41740) was investigated for its ability to induce the expression of cytokine mRNAs in murine peritoneal macrophages in vitro. Northern blot analysis showed that in quiescent macrophage populations, both IL-1 alpha and IL-1 beta mRNA levels were dramatically increased in response to 1 microgram/ml Biostim. Dot-blot analysis showed that in quiescent macrophage populations the expression of mRNAs for IL-1 alpha, IL-1 beta, IL-6 and TNF-alpha could be elevated by concentrations of Biostim as low as 1-10 pg/ml, detectable after 3 h exposure. In parallel experiments LPS was effective only at the higher concentration of 10 ng/ml. Time-course analysis showed that the expression of these cytokine mRNAs was transient, peaking after 1-3 h; only transcripts of IL-1 beta were detectable after 23 h exposure. No effects were seen on the expression of actin, a high-turnover housekeeping gene. We propose that this type of analysis represents a sensitive, specific and reproducible method for assessing the ability of drugs and chemicals to modulate the expression of cytokines that play a pivotal role in the induction of the immune response.

Animals↗
PubMed1989

Species distribution of paraoxon-resistant brain polypeptides radiolabelled with diisopropyl phosphorofluoridate ([3H]DiPF): electrophoretic assay for the aged polypeptide of [3H]DiPF-labelled neuropathy target esterase.

Brain neuropathy target esterase is identified as a paraoxon-resistant, mipafox-sensitive esterase that can be labelled with [3H]diisopropyl phosphorofluoridate. During "aging" of the labelled (inhibited) esterase, half the label (one isopropyl group) is transferred to a site (of the same molecular weight in sodium dodecyl sulphate) whence it may be released in volatile form by treatment with alkali. Our previously published procedure for complete extraction in a form suitable for scintillation counting of tritium-labelled proteins from polyacrylamide gels includes treatment of part-solubilised gels with alkali. Particles from brain of the hen, pig, sheep, guinea-pig, and rat were preincubated with paraoxon with or without mipafox, treated with [3H]diisopropyl phosphorofluoridate, and solubilised in sodium dodecyl sulphate. Labelled polypeptides (except from the rat) were separated by electrophoresis. Both mipafox-sensitive labelling and "volatilisable counts" were located principally in the 155-kilodalton region, with the residues dispersed throughout the gels. The quantities of paraoxon-resistant, mipafox-sensitive labelling sites and of "volatilisable counts" (in pmol/particles from 1 g) were, respectively, 12.2 and 8.65 in hen brain, 9.80 and 6.82 in pig, 8.48 and 5.46 in sheep, 4.46 and 4.01 in guinea-pig, and 4.91 and 2.08 in rat. The "volatilisable count" assay seems more specific for neuropathy target esterase and is easier and more precise than assays based on differences in labelling of two samples, each subjected to much processing. Hydrolytic activity of particles taken before labelling was measured against phenyl valerate.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1988

Phenyl di-n-pentylphosphinate: a convenient reactivatible inhibitor for studies on neuropathy target esterase (NTE) and protection against organophosphate-induced delayed polyneuropathy.

Phenyl di-n-pentylphosphinate was synthesised by interaction of phenyl phosphorodichloridate and n-pentyl magnesium bromide. The product was purified by silica chromatography (yield 25%). Although much more stable at physiological pH than its 4-nitrophenyl analogue, this ester is a good inhibitor of neuropathy target esterase (NTE): kappa a = 1.7 X 10(5) M-1 min-1. It is a very weak anticholinesterase (kappa a congruent to 10 M-1 min-1). In vivo only 5-10 mg/kg is required to inhibit hen brain and spinal cord NTE. The inhibited NTE can be reactivated fully by incubation in vitro with iso-nitrosoacetophenone (INAP) (19 mM at 37 degrees C and pH 8.5 for 60 min): this property enables study to be made of the fate of inhibited NTE in vivo.

Animals↗
PubMed1988

Neuropathy target esterase: rates of turnover in vivo following covalent inhibition with phenyl di-n-pentylphosphinate.

Phenyl di-n-pentylphosphinate is a reasonably stable easily synthesized inhibitor of neuropathy target esterase (NTE) with low anticholinesterase activity. Like phenylmethylsulphonyl fluoride it protects hens against neuropathic effects of compounds such as diisopropylphosphorofluoridate. At intervals up to 15 days after dosing hens (10 mg/kg s.c. to inhibit 90% NTE) assays were made of catalytically active and of phosphinylated NTE in autopsy tissue. The sum of these components was always within the range of catalytic activity in undosed controls. However, the half-life of reappearance of active NTE was 2.07 days +/- 0.13 (SD, n = 6) for brain and 3.62 days +/- 0.23 (SD, n = 6) for spinal cord--shorter than after dosing with phenylmethylsulphonyl fluoride. It is proposed that: (1) The physiological turnover mechanism cannot distinguish between catalytically active and di-n-pentylphosphinylated NTE although initiation of organophosphate-induced delayed polyneuropathy might involve recognition of aged di-alkyl-phosphorylated NTE as "foreign". (2) The short half-lives indicate a slow spontaneous dephosphinylation of inhibited NTE occurs in vivo as well as de novo synthesis. The difference in half-lives for brain and spinal cord NTE may be due to different rates of synthesis de novo or (more likely) to different rates of spontaneous reactivation of the inhibited NTE in the two tissues.

Animals↗
PubMed1987

Complete extraction in a form suitable for liquid scintillation counting of tritium-labeled proteins from polyacrylamide gels.

Large (200 mm3) slices of polyacrylamide gels crosslinked with N,N'-diallyltartardiamide which contain tritium-labeled protein are readily solubilized in periodic acid for liquid scintillation counting of radioactivity, but the apparent recovery of label never exceeds 82%. Extraction of the slices with two commercial solubilizers at 60 degrees C gave recoveries of 82-90% which were not improved by prolonged incubation. Treatment of the slices at ambient temperature with 1.0 ml of 2% sodium periodate for 30 min followed by the addition of 0.7 ml of aqueous tetrabutylammonium hydroxide (40% w/v) gives solutions which can be immediately counted at 35% efficiency with low background and with 100% recovery of tritiated protein

Electrophoresis, Polyacrylamide Gel↗
PubMed1987

Influence of a 3.5 day fast on physical performance.

Eight young men were tested for strength, anaerobic capacity and aerobic endurance in a post absorptive state and after a 3.5 day fast. Strength was tested both isokinetically (elbow flexors, 0.52 rad x s-1 and 3.14 rad x s-1) and isometrically. Anaerobic capacity was evaluated by having subjects perform 50 rapidly repeated isokinetic contractions of the elbow flexors at 3.14 rad x s-1. Aerobic endurance was measured as time to volitional fatigue during a cycle ergometer exercise at 45% VO2max. Measures of VO2, VE, heart rate, and ratings of perceived exertion were obtained prior to and during the cycle exercise. The 3.5 day fast did not influence isometric strength, anaerobic capacity or aerobic endurance. Isokinetic strength was significantly reduced (approximately 10%) at both velocities. VO2, VE and perceived exertion were not affected by fasting. Fasting significantly increased heart rate during exercise but not at rest. It was concluded that there are minimal impairments in physical performance parameters measured here as a result of a 3.5 day fast.

Adult↗
PubMed1986

Isolation and properties of the signal region from ovalbumin.

A tryptic fragment (residues 21-47) containing the signal peptide of ovalbumin has been isolated by reverse-phase chromatography. The peptide is more active at inhibiting the processing of pre-prolactin in an in vitro translation system than fragments of ovalbumin isolated previously, and is similar in properties to synthetic signal peptides. The ovalbumin signal fragment is shown to bind to a protein component of salt-stripped pancreatic microsomal membranes, which is cross-linked under UV irradiation by a radioactive synthetic photoaffinity signal peptide probe to yield a radiolabelled 45 kDa protein.

Amino Acid Sequence↗