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D H Powell

Publications and source records attributed to D H Powell.

14 recordsLinked to original sources
PubMed2001

Capillary LC-MS2 at the attomole level for monitoring and discovering endogenous peptides in microdialysis samples collected in vivo.

Fused-silica capillary LC columns (25-microm i.d.) with 3-microm-i.d. integrated electrospray emitters interfaced to a quadrupole ion trap mass spectrometer were evaluated for high-sensitivity LC-MS2. Column preparation involved constructing frits by in situ photopolymerization of glycidyl methacrylate and trimethylolpropane trimethacrylate, preparing the electrospray emitter by pulling the column outlet to a fine tip with a CO2 laser puller, and slurry-packing the column with 5-microm reversed-phase particles. Large-volume injections were facilitated by an automated two-pump system that allowed high-flow rates for sample loading and low-flow rates for elution. Small electrospray emitters, low elution flow rates, and optimization of gradient steepness allowed a detection limit of 4 amol, corresponding to 2 pM for 1.8 microL injected on-column, for a mixture of peptides dissolved in artificial cerebral spinal fluid. The system was coupled on-line to microdialysis sampling and was used to monitor and discover endogenous neuropeptides from the globus pallidus of anesthetized male Sprague-Dawley rats. Time-segmented MS2 scans enabled simultaneous monitoring of Met-enkephalin, Leu-enkephalin, and unknown peptides. Basal dialysate levels of Met-enkephalin and Leu-enkephalin were 60 +/- 30 and 70 +/- 20 pM while K+-stimulated levels were 1,900 +/- 500 and 1,300 +/- 300 pM, respectively (n = 7). Data-dependent and time-segmented MS2 scans revealed several unknown peptides that were present in dialysate. One of the unknowns was identified as peptide I(1-10) (SPQLEDEAKE), a novel product of preproenkephalin A processing, using MS2, MS3, and database searching.

Amino Acid Sequence↗
PubMed2001

First solvation shell of the Cu(II) aqua ion: evidence for fivefold coordination.

We determined the structure of the hydrated Cu(II) complex by both neutron diffraction and first-principles molecular dynamics. In contrast with the generally accepted picture, which assumes an octahedrally solvated Cu(II) ion, our experimental and theoretical results favor fivefold coordination. The simulation reveals that the solvated complex undergoes frequent transformations between square pyramidal and trigonal bipyramidal configurations. We argue that this picture is also consistent with experimental data obtained previously by visible near-infrared absorption, x-ray absorption near-edge structure, and nuclear magnetic resonance methods. The preference of the Cu(II) ion for fivefold instead of sixfold coordination, which occurs for other cations of comparable charge and size, results from a Jahn-Teller destabilization of the octahedral complex.

Journal Article↗
PubMed2001

Integration of mass spectrometry into early-phase discovery and development of central nervous system agents.

The early-phase discovery and development of useful central nervous system (CNS) agents present ample opportunities to exploit mass spectrometry and provide detailed compound/mixture characterization, or to make the process faster and/or more economic. Neuropeptide FF antagonists and centrally active thyrotropin-releasing hormone analogues were used as specific examples in this work. We evaluated the characterization of focused libraries of peptide derivatives by electrospray ionization, tandem mass spectrometry and liquid chromatography/tandem mass spectrometry on a quadrupole ion trap and nanoelectrospray on a Fourier transform ion cyclotron resonance mass spectrometer. Immobilized artificial-membrane chromatography was employed as a model to predict/rank new agents against lead compounds for their potential to reach the central nervous system in pharmacologically significant amounts. Measuring brain concentrations in rodents after the intravenous administration of test compounds was used as an in vivo approach, and we took advantage of microdialysis sampling that furnished samples without interfering tissue matrix and afforded the estimation of extracellular concentrations in a localized part of the brain. Overall, making atmospheric-pressure ionization mass spectrometry an integral part of the process has played a major role in increasing throughput, selectivity, specificity and detection sensitivity and thereby providing useful information about the extent or mechanism of transport and metabolic activation/inactivation in early-phase discovery and development of CNS agents.

Animals↗
PubMed2000

The missing organic molecules on Mars.

GC-MS on the Viking 1976 Mars missions did not detect organic molecules on the Martian surface, even those expected from meteorite bombardment. This result suggested that the Martian regolith might hold a potent oxidant that converts all organic molecules to carbon dioxide rapidly relative to the rate at which they arrive. This conclusion is influencing the design of Mars missions. We reexamine this conclusion in light of what is known about the oxidation of organic compounds generally and the nature of organics likely to come to Mars via meteorite. We conclude that nonvolatile salts of benzenecarboxylic acids, and perhaps oxalic and acetic acid, should be metastable intermediates of meteoritic organics under oxidizing conditions. Salts of these organic acids would have been largely invisible to GC-MS. Experiments show that one of these, benzenehexacarboxylic acid (mellitic acid), is generated by oxidation of organic matter known to come to Mars, is rather stable to further oxidation, and would not have been easily detected by the Viking experiments. Approximately 2 kg of meteorite-derived mellitic acid may have been generated per m(2) of Martian surface over 3 billion years. How much remains depends on decomposition rates under Martian conditions. As available data do not require that the surface of Mars be very strongly oxidizing, some organic molecules might be found near the surface of Mars, perhaps in amounts sufficient to be a resource. Missions should seek these and recognize that these complicate the search for organics from entirely hypothetical Martian life.

Alkanes↗
PubMed1996

Auditory and visual recall of forward and backward digit spans.

Increasingly, clinicians obtain data from tests across modalities. Little actual information, however, exists as to normal patterns. An example is auditory and visual recall of digit span. The effects of modality on recall of digit span, sequence, and order were examined with 80 subjects (M age 24.3 yr.). Subjects were given the Wechsler Adult Intelligence Scale-Revised, and a research version of MicroCog: Assessment of Cognitive Functioning. Both contain forward and backward digit spans. Recall for visual presentation was significantly stronger than for auditory presentation of backward digit span. Recall for visual backward digit span was also superior to auditory backward digit span when the sequence of presentation was controlled. The mean number of digits recalled was higher when visual digit span was preceded by auditory digit span.

Adult↗
PubMed1993

Steroidogenesis in Fundulus heteroclitus V.: purification, characterization, and metabolism of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one by intact follicles and its role in oocyte maturation.

In vitro steroidogenesis of ovarian follicles incubated with radioactive precursors or a Fundulus heteroclitus pituitary extract (FPE) was investigated. Steroids were extracted from both the medium and follicular tissue and fractionated by liquid or thin-layer chromatography. A similar pattern of steroid metabolites was obtained with either [14C]pregnenolone or [14C]progesterone as exogenous precursor. Several metabolites comigrated with known reference steroids and thus were tentatively identified. Some have been previously reported to induce germinal vesicle breakdown (GVBD) in this and other species, particularly 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (DHP). [3H]DHP added to intact follicles or denuded oocytes was also extensively metabolized. All the DHP metabolites produced by the intact follicle were tested for biological activity. Three of the metabolites were almost as effective inducers of GVBD as DHP itself, and two were tentatively identified as 5 alpha-pregnan-3 alpha,17 alpha,20 beta-triol and 5 alpha-pregnan-3 beta,17 alpha,20 beta-triol. However, DHP was the most potent and the quickest inducer of GVBD, indicating that its maturational action is not due to metabolic conversion to a more active form. In addition, we found two very active fractions after HPLC analysis of steroid extracts from FPE-stimulated follicles: one that corresponded to and was further identified (mass spectroscopy) as DHP and a second tentatively identified as the DHP metabolite 5 alpha-pregnan-3 beta,17 alpha,20 beta-triol. This study provides strong evidence that DHP plays the major role as a maturation inducing-steroid in F. heteroclitus, even though DHP is not the only active steroid produced by maturing follicles.

Animals↗
PubMed1976

Radiation-induced peritoneal mesothelioma.

A case report of a patient who developed peritoneal mesothelioma 7 years after internal and external irradiation for carcinoma of the cervix is reported. No previous reports of induction of this tumor by irradiation have been found. The subject of radiation-induced tumors and peritoneal mesothelioma is briefly discussed.

Female↗
PubMedDate not supplied

Bioconjugation of ribonuclease A: a detailed chromatographic and mass spectrometric analysis of chemical modification by a cross-linking reagent.

The modification of ribonuclease A with the heterobifunctional cross-linker, 4-succinimdyloxycarbonyl-methyl-alpha-[2-pyridyldithio]-toluene (SMPT) is described. RNase A has 11 potential sites of modification by the SMPT reagent. Tracking the two-dimensional separation and proteolytic digestion of SMPT-modified RNase A with ESI/FTICR-MS and HPLC/ESI/QIT-MS demonstrates the detailed information about number of SMPT modifications and sites of modification that can be obtained by application of these techniques. Analysis of native and modified RNase A tryptic digests by ESI/FTICR-MS resulted in the identification of the sites of modification. Semiquantitative results of the reactivity of certain lysine residues toward the coupling reagent SMPT are presented. Two sites (lysines 1 and 37) are highly reactive, while three sites (lysines 41, 61, and 104) appear to be unreactive toward SMPT under the conditions used. Experimental results demonstrate that quantitative comparison of relative intensities of peptide sequences of different charge states is not possible. No correlation was found between number of basic residues and sensitivity to detection. Digestion of the modified and unmodified RNase A by subtilisin followed by examination by HPLC/ESI/QIT-MS and MS(n) enabled further investigation of modification on lysines 1 and 7, including modification at the epsilon- and alpha-amino positions on lysine 1.

Amino Acid Sequence↗