Biomedical subjects

Keith W Jones

Publications and source records attributed to Keith W Jones.

21 records · Page 2Linked to original sources

PubMed2002

The CRF(1) receptor antagonist DMP696 produces anxiolytic effects and inhibits the stress-induced hypothalamic-pituitary-adrenal axis activation without sedation or ataxia in rats.

RATIONALE: CRF(1) antagonists may be effective in the treatment of anxiety disorders while having fewer side effects compared with classical benzodiazepines. OBJECTIVES: The effects of a small molecule selective CRF(1) antagonist DMP696 on anxiety-like behaviors and stress-induced increases in corticosterone in rats exposed to a novel environment and on locomotor activity and motor coordination were determined in rats. These effects of DMP696 were compared with those produced by the classical benzodiazepine chlordiazepoxide (CDP). METHODS: DMP696 or CDP were administered PO, 60 minutes before behavioral testing in rats. Their effects on latency to exit a dark chamber and stress-induced increase in corticosterone in the Defensive Withdrawal test (an animal model of anxiety), locomotor activity, and rotorod performance (measure of ataxia) were determined. RESULTS: DMP696 significantly reduced exit latency and reversed the stress-induced increase in corticosterone in the Defensive Withdrawal test at doses of 3.0-10 mg/kg and higher. In contrast, CDP significantly decreased exit latency at 10 and 30 mg/kg, but not at 100 mg/kg, due to concurrent non-specific side effects. Unlike DMP696, CDP had no effect on the stress-induced increase in corticosterone at lower doses, but resulted in a significant increase at higher doses. DMP696 did not reduce locomotor activity or impair motor coordination at doses up to 30-fold higher than doses effective in the Defensive Withdrawal model. In contrast, CDP produced significant sedation and ataxia at the same doses that were effective in reducing exit latency. CONCLUSIONS: These data suggest that the CRF(1) antagonist DMP696 might retain the therapeutic benefits of classical benzodiazepines but have fewer motoric side effects.

Adrenocorticotropic Hormone↗

PubMed2002

Calcium compartments in brain.

Excellent progress has been made toward understanding the physiology and pharmacology of specific calcium-related cellular processes of the brain, but few studies have provided an integrated view of brain calcium kinetics. To further the knowledge of the size and binding properties of brain calcium compartments, the authors have conducted a series of experiments in hippocampal brain slices exposed to high and low extracellular calcium. Slices were incubated in buffers containing 0.001 to 4.5 mmol/L calcium for up to 75 minutes. Slice calcium content was analyzed by three methods: exchange equilibrium with 45Ca, synchrotron-radiation-induced x-ray emission, and inductively coupled plasma. Data were analyzed using a model based on a Langmuir isotherm for two independent sites, with additional extracellular and bound compartments. In parallel experiments, altered low calcium had no effect on slice histology and only mild effects on slice adenylates. When combined with prior 45Ca and fluorescent probe binding experiments, these results suggest that there are at least five kinetically distinct calcium compartments: (1) free extracellular (approximately 10%); (2) loosely associated extracellular plasma membrane (approximately 55%); (3) intracellular compartment with moderate avidity (approximately 17%); (4) tightly bound, nonexchangeable intracellular compartment ( approximately 15%); and (5) free cytoplasmic (<0.01%). If only the third compartment is considered a potential calcium buffer, then the buffering ratio is calculated to be approximately 2,700:1, but if the second compartment is also included, then the buffering ratio would be approximately 13,000:1. This may explain the wide range of estimates observed by fluorescent probe studies.

Animals↗

PubMed2002

Synchrotron radiation-induced X-ray emission to identify metal ions in preparations of purified protein.

The suitability of synchrotron radiation-induced X-ray emission (SRIXE) for the detection and identification of metal ions in preparations of purified, soluble proteins was tested. Glutathione S-transferase fused to the proximal zinc finger motif of human transcription factor IIIA or to the cysteine-rich motif of poliovirus protein 2C was expressed in bacteria and purified by affinity chromatography. Aqueous samples containing the purified proteins were analyzed with SRIXE, and trace amounts of zinc and iron were detected. Mutation of the zinc-coordinating residues in the cysteine-rich motif of poliovirus protein 2C resulted in the loss of the zinc-binding ability. Relative quantities of metal in the protein preparations as determined by SRIXE corresponded well with the metal:protein ratios calculated by using a 4-(2-pyridylazo)resorcinol-based assay. We conclude that SRIXE is an accurate, sensitive, and simple method for the detection and identification of protein-bound metal ions in small amounts of sample. Thus, SRIXE may have wide use as a particularly effective method for rapidly determining trace metals in microarray samples.

Amino Acid Sequence↗