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Biomedical subjects

R Powers

Publications and source records attributed to R Powers.

At least 37 records · Page 2Linked to original sources
PubMed2000

Lead exposure from blood transfusion to premature infants.

OBJECTIVE: The objective of this study was to determine the exposure of premature infants to lead from blood transfusions. STUDY DESIGN: Blood lead concentrations were determined for 19 very premature infants at the time of admission, at 4 weeks of life, and before and after transfusions and in the donor packed red blood cells of 79 transfusions. RESULTS: The number of transfusions per patient was 4. 2 +/- 2.8 (mean +/- SD) with 15.7 +/- 1.9 mL/kg packed red blood cells for a lead dose of 1.56 +/- 1.77 microg/dL. The total dose of lead from these transfusions over the 4-week period was 4.0 +/- 2.8 microg/kg (range, 0.9-10.6 microg/kg). Increases in post-transfusion blood lead concentration were linear with doses higher than 1.5 microg/dL. Packed red blood cells with a blood lead concentration of > or = 5 microg/dL resulted in an elevated post-transfusion blood lead concentration in some infants. CONCLUSIONS: The lead exposure to these infants through blood transfusion exceeds the acceptable daily intake values for lead and may result in unacceptably high post-transfusion blood lead concentrations. Use of packed red blood cells with lead concentrations <3.3 microg/dL is one cost-effective means to reduce exposure.

Blood Transfusion↗
PubMed1999

NMR solution structure of the catalytic fragment of human fibroblast collagenase complexed with a sulfonamide derivative of a hydroxamic acid compound.

The solution structure of the catalytic fragment of human fibroblast collagenase (MMP-1) complexed with a sulfonamide derivative of a hydroxamic acid compound (CGS-27023A) has been determined using two-dimensional and three-dimensional heteronuclear NMR spectroscopy. The solution structure of the complex was calculated by means of hybrid distance geometry-simulated annealing using a combination of experimental NMR restraints obtained from the previous refinement of the inhibitor-free MMP-1 (1) and recent restraints for the MMP-1:CGS-27023A complex. The hydroxamic acid moiety of CGS-27023A was found to chelate to the "right" of the catalytic zinc where the p-methoxyphenyl sits in the S1' active-site pocket, the isopropyl group is in contact with H83 and N80, and the pyridine ring is solvent exposed. The sulfonyl oxygens are in hydrogen-bonding distance to the backbone NHs of L81 and A82. This is similar to the conformation determined by NMR of the inhibitor bound to stromelysin (2, 3). A total of 48 distance restraints were observed between MMP-1 and CGS-27023A from 3D 13C-edited/12C-filtered NOESY and 3D 15N-edited NOESY experiments. An additional 18 intramolecular restraints were observed for CGS-27023A from a 2D 12C-filtered NOESY experiment. A minimal set of NMR experiments in combination with the free MMP-1 assignments were used to assign the MMP-1 (1)H, 13C, and 15N resonances in the MMP-1:CGS-27023A complex. The assignments of CGS-27023A in the complex were obtained from 2D 12C-filtered NOESY and 2D 12C-filtered TOCSY experiments.

Catalysis↗
PubMed1999

HYPER: a hierarchical algorithm for automatic determination of protein dihedral-angle constraints and stereospecific C beta H2 resonance assignments from NMR data.

A new computer program, HYPER, has been developed for automated analysis of protein dihedral angle values and C beta H2 stereospecific assignments from NMR data. HYPER uses a hierarchical grid-search algorithm to determine allowed values of phi, psi, and chi 1 dihedral angles and C beta H2 stereospecific assignments based on a set of NMR-derived distance and/or scalar-coupling constraints. Dihedral-angle constraints are valuable for restricting conformational space and improving convergence in three-dimensional structure calculations. HYPER computes the set of phi, psi, and chi 1 dihedral angles and C beta H2 stereospecific assignments that are consistent with up to nine intraresidue and sequential distance bounds, two pairs of relative distance bounds, thirteen homo- and heteronuclear scalar coupling bounds, and two pairs of relative scalar coupling constant bounds. The program is designed to be very flexible, and provides for simple user modification of Karplus equations and standard polypeptide geometries, allowing it to accommodate recent and future improved calibrations of Karplus curves. The C code has been optimized to execute rapidly (0.3-1.5 CPU-sec residue-1 using a 5 degrees grid) on Silicon Graphics R8000, R10000 and Intel Pentium CPUs, making it useful for interactive evaluation of inconsistent experimental constraints. The HYPER program has been tested for internal consistency and reliability using both simulated and real protein NMR data sets.

Algorithms↗
PubMed1998

Homology model for oncostatin M based on NMR structural data.

Oncostatin M (OM) is a member of the cytokine family which regulates the proliferation and differentiation of a variety of cell types and includes interleukin-6 (IL-6), leukemia inhibitory factor (LIF), and granulocyte-colony stimulating factor (G-CSF). This family of proteins adopts a four-helix bundle fold with up-up-down-down topology and contains intramolecular disulfide bonds. Since an X-ray or NMR structure for OM is not currently available, a homology model for OM was determined from the X-ray structures of human growth hormone (hGH), LIF, and G-CSF where the alignment was based on secondary structure instead of sequence. The OM secondary structure was determined from NMR structural data, and the secondary structures for hGH, LIF, and G-CSF were obtained from the reported X-ray structures. The resulting homology model was refined using sequential NOE distance 13C restraints, chemical shift information, and a conformational database.

Amino Acid Sequence↗
PubMed1998

High-resolution solution structure of the inhibitor-free catalytic fragment of human fibroblast collagenase determined by multidimensional NMR.

The high-resolution solution structure of the inhibitor-free catalytic fragment of human fibroblast collagenase (MMP-1), a protein of 18.7 kDa, which is a member of the matrix metalloproteinase family, has been determined using three-dimensional heteronuclear NMR spectroscopy. A total of 30 structures were calculated by means of hybrid distance geometry-simulated annealing using a total of 3333 experimental NMR restraints, consisting of 2409 approximate interproton distance restraints, 84 distance restraints for 42 backbone hydrogen bonds, 426 torsion angle restraints, 125 3JNH alpha restraints, 153 C alpha restraints, and 136 C beta restraints. The atomic rms distribution about the mean coordinate positions for the 30 structures for residues 7-137 and 145-163 is 0.42 +/- 0.04 A for the backbone atoms, 0.80 +/- 0.04 A for all atoms, and 0.50 +/- 0.03 A for all atoms excluding disordered side chains. The overall structure of MMP-1 is composed of a beta-sheet consisting of five beta-strands in a mixed parallel and anti-parallel arrangement and three alpha-helices. A best-fit superposition of the NMR structure of inhibitor-free MMP-1 with the 1.56 A resolution X-ray structure by Spurlino et al. [Spurlino, J. C., Smallwood, A. M., Carlton, D. D., Banks, T. M., Vavra, K. J., Johnson, J. S., Cook, E. R., Falvo, J., and Wahl, R. C., et al. (1994) Proteins: Struct., Funct., Genet. 19, 98-109] complexed with a hydroxamate inhibitor yields a backbone atomic rms difference of 1.22 A. The majority of differences between the NMR and X-ray structure occur in the vicinity of the active site for MMP-1. This includes an increase in mobility for residues 138-144 and a displacement for the Ca(2+)-loop (residues 74-80). Distinct differences were observed for side-chain torsion angles, in particular, the chi 1 for N80 is -60 degrees in the NMR structure compared to 180 degrees in the X-ray. This results in the side chain of N80 occupying and partially blocking access to the active site of MMP-1.

Binding Sites↗
PubMed1998

Concomitant rash and blood dyscrasias in geriatric psychiatry patients treated with carbamazepine.

BACKGROUND: Rashes and blood dyscrasias are disconcerting adverse effects associated with carbamazepine therapy. Rashes are quite common, as are mild blood dyscrasias, such as mild leukopenias. Fortunately, severe rashes and blood dyscrasias are rare. There are few reports on the relationship between carbamazepine-induced rashes and blood dyscrasias, including a prospective study in which rash appeared concomitantly with leukopenia and/or thrombocytopenia in 10 patients, two case reports in which simultaneous rash and agranulocytosis occurred, and two case reports in which rashes served as harbingers of fatal aplastic anemia. CASE REPORTS: We report two cases of concomitant rashes and blood dyscrasias in geriatric psychiatry patients receiving carbamazepine therapy for bipolar disorder. One patient was found to have a severe leukopenia within several days after rash onset. The other patient was discovered to have a severe leukopenia and thrombocytopenia within about a month after rash onset. DISCUSSION: Current hematologic monitoring guidelines for carbamazepine rely heavily on the recognition of signs and symptoms of blood dyscrasias by clinicians and patients. We believe that our cases support the suggestion that patients who develop rashes receive more vigilant monitoring of the complete blood count, should carbamazepine therapy by continued. Given the currently available case reports and the fact that the incidence of drug-induced blood dyscrasias increases with advanced age, this recommendation may be particularly relevant for geriatric patients. CONCLUSIONS: Further study is required to establish whether carbamazepine-induced concomitant rashes and blood dyscrasias are valid associations insofar as monitoring is concerned.

Aged↗
PubMed1998

The oral health status of individuals on renal dialysis.

The primary purpose of this investigation was to determine the oral health status of individual's undergoing renal dialysis in southeastern Virginia. A sample was identified for this cross-sectional study via a local health maintenance organization. Three subgroups of the population were studied: 1) those who have been on renal dialysis for less than a year; 2) those on renal dialysis for 1 to 3 years; and 3) those on renal dialysis for longer than 3 years. Three dental indices the Periodontal Disease Index; the Decayed, Missing. Filled index; and the Simplified Oral Hygiene Index--were used to identify periodontal disease, dental caries activity, and oral hygiene status. Data were compiled and analyzed by using the parametric test, 1-way analysis of variance. Results suggested that 100% (n = 45) of the individuals undergoing renal dialysis presented with some form of periodontal disease (X = 3.15, SD = 1.41). The majority (64%) of the sample displayed either severe gingivitis (28%) or early periodontitis (36%). Sixty-four percent of the sample displayed a high DMF index (X = 11.77, SD = 7.55), while 98% of the sample accumulated calculus. Oral debris was present in 100% of the sample, resulting in a high Simplified Oral Hygiene Index score (X = 3.24, SD = 1.26), suggesting an increased need for oral care instruction. Findings led to the conclusion that the renal dialysis population in southeastern Virginia, regardless of length of time on dialysis, is in need of comprehensive professional oral care and self-care instruction. Oral disease was present and is a source of active infection in these medically compromised individuals and, as such, has dire implication for morbidity and mortality.

Analysis of Variance↗
PubMed1998

Opioid withdrawal in neonates after continuous infusions of morphine or fentanyl during extracorporeal membrane oxygenation.

BACKGROUND: Complications of opioid analgesia include tolerance and withdrawal. OBJECTIVES: To determine the effects of morphine and fentanyl on the prevalence of withdrawal after extracorporeal membrane oxygenation. METHODS: Two groups of neonates were compared during and after extracorporeal membrane oxygenation: a prospective group receiving a continuous infusion of morphine for analgesia and sedation and a retrospective group who had received a continuous infusion of fentanyl. RESULTS: Neonates receiving morphine required significantly less supplemental analgesia (P < .001) than did neonates who had received fentanyl and had a significantly lower prevalence of withdrawal after the therapy (P = .01). Neonates receiving morphine were discharged from the hospital a mean of 9.6 days sooner (P = .01) than neonates who had received fentanyl. CONCLUSIONS: Morphine may offer marked advantages over fentanyl for providing continuous analgesia and sedation in neonates.

Analgesics, Opioid↗
PubMed1997

Automated analysis of protein NMR assignments using methods from artificial intelligence.

An expert system for determining resonance assignments from NMR spectra of proteins is described. Given the amino acid sequence, a two-dimensional 15N-1H heteronuclear correlation spectrum and seven to eight three-dimensional triple-resonance NMR spectra for seven proteins, AUTOASSIGN obtained an average of 98% of sequence-specific spin-system assignments with an error rate of less than 0.5%. Execution times on a Sparc 10 workstation varied from 16 seconds for smaller proteins with simple spectra to one to nine minutes for medium size proteins exhibiting numerous extra spin systems attributed to conformational isomerization. AUTOASSIGN combines symbolic constraint satisfaction methods with a domain-specific knowledge base to exploit the logical structure of the sequential assignment problem, the specific features of the various NMR experiments, and the expected chemical shift frequencies of different amino acids. The current implementation specializes in the analysis of data derived from the most sensitive of the currently available triple-resonance experiments. Potential extensions of the system for analysis of additional types of protein NMR data are also discussed.

Automation↗
PubMed1997

Properly oriented heparin-decasaccharide-induced dimers are the biologically active form of basic fibroblast growth factor.

Interaction of basic fibroblast growth factor (FGF-2) with heparin or heparan sulfate proteoglycans (HSPGs) is required for receptor activation and initiation of biological responses. To gain insight into the mechanism of activation of the FGF receptor by FGF-2 and heparin, we have used NMR, dynamic light scattering, and HSPG-deficient cells and cell-free systems. The first 28 N-terminal residues in FGF-2 were found to be highly mobile and flexible, consistent with the disorder found in both the NMR and X-ray structures. The structure of an FGF-2-heparin-decasaccharide complex that binds to and activates the FGF receptor was compared to a heparin-tetrasaccharide-induced complex that does not promote an interaction with the receptor. The major change observed upon the addition of the tetrasaccharide to FGF-2 was an increase in the correlation time consistent with the formation of an FGF-2 dimer. The NMR line widths of FGF-2 in the presence of the decasaccharide are severely broadened relative to the tetrasaccharide, consistent with dynamic light scattering results which indicate FGF-2 is a tetramer. The interaction of these heparin species with FGF-2 does not induce a significant conformational change in the overall structure of FGF-2, but small chemical shift changes are observed in both heparin and receptor binding sites. A trans-oriented symmetric dimer of FGF-2 is formed in the presence of the tetrasaccharide whereas two cis-oriented dimers in a symmetric tetramer are formed in the presence of the decasaccharide. This suggests that the cis-oriented FGF-2 dimer is the minimal biologically active structural unit of FGF-2. These data allow us to propose a novel mechanism to explain the functional interaction of FGF-2 with heparin and its transmembrane receptor.

Animals↗
PubMed1997

Assignments, secondary structure and dynamics of the inhibitor-free catalytic fragment of human fibroblast collagenase.

Fibroblast collagenase (MMP-1), a 169-residue protein with a molecular mass of 18.7 kDa, is a matrix metalloproteinase which has been associated with pathologies such as arthritis and cancer. The assignments of the 1H, 15N, 13CO and 13C resonances, determination of the secondary structure and analysis of 15N relaxation data of the inhibitor-free catalytic fragment of recombinant human fibroblast collagenase (MMP-1) are presented. It is shown that MMP-1 is composed of a beta-sheet consisting of five beta-strands in a mixed parallel and antiparallel arrangement (residues 13-19, 48-53, 59-65, 82-85 and 94-99) and three alpha-helices (residues 27-43, 112-124 and 150-160). This is nearly identical to the secondary structure determined from the refined X-ray crystal structures of inhibited MMP-1. The major difference observed between the NMR solution structure of inhibitor-free MMP-1 and the X-ray structures of inhibited MMP-1 is the dynamics of the active site. The 2D 15N-1H HSQC spectra, the lack of information in the 15N-edited NOESY spectra, and the generalized order parameters (S2) determined from 15N T1, T2 and NOE data suggest a slow conformational exchange for residues comprising the active site (helix B, zinc ligated histidines and the nearby loop region) and a high mobility for residues Pro138-Gly144 in the vicinity of the active site for inhibitor-free collagenase. In contrast to the X-ray structures, only the slow conformational exchange is lost in the presence of an inhibitor.

Amino Acid Sequence↗
PubMed1996

High-resolution solution structure of basic fibroblast growth factor determined by multidimensional heteronuclear magnetic resonance spectroscopy.

The high-resolution solution structure of recombinant human basic fibroblast growth factor (FGF-2), a protein of 17.2 kDa that exhibits a variety of functions related to cell growth and differentiation, has been determined using three-dimensional heteronuclear NMR spectroscopy. A total of 30 structures were calculated by means of hybrid distance geometry--simulated annealing using a total of 2865 experimental NMR restraints, consisting of 2486 approximate inteproton distance restraints, 50 distance restraints for 25 backbone hydrogen bonds, and 329 torsion angle restraints. The atomic rms distribution about the mean coordinate positions for the 30 structures for residues 29-152 is 0.43 +/- 0.03 A for the backbone atoms, 0.83 +/- 0.05 A for all atoms, and 0.51 +/- 0.04 A for all atoms excluding disordered side chains. The overall structure of FGF-2 consists of 11 extended antiparallel beta-strands arranged in three groups of three or four strands connected by tight turns and loop regions creating a pseudo-3-fold symmetry. Two strands from each group come together to form a beta-sheet barrel of six antiparallel beta-strands. A helix-like structure was observed for residues 131-136, which is part of the heparin binding site (residues 128-138). The discovery of the helix-like region in the primary heparin binding site instead of the beta-strand conformation described in the X-ray structures may have important implications in understanding the nature of heparin--FGF-2 interactions. A total of seven tightly bound water molecules were found in the FGF-2 structure, two of which are located in the heparin binding site. The first 28 N-terminal residues appear to be disordered, which is consistent with previous X-ray structures. A best fit superposition of the NMR structure of FGF-2 with the 1.9 A resolution X-ray structure by Zhu et al. (1991) yields a backbone atomic rms difference of 0.94 A, indicative of a close similarity between the NMR and X-ray structures.

Binding Sites↗
PubMed1996

Resonance assignments for Oncostatin M, a 24-kDa alpha-helical protein.

Oncostatin M (OM) is a cytokine that shares a structural and functional relationship with interleukin-6, leukemia inhibitory factor, and granulocyte-colony stimulating factor, which regulate the proliferation and differentiation of a variety of cell types. A mutant version of human OM in which two N-linked glycosylation sites and an unpaired cysteine have been mutated to alanine (N76A/C81A/N193A) has been expressed and shown to be active. The triple mutant has been doubly isotope-labeled with 13C and 15N in order to utilize heteronuclear multidimensional NMR techniques for structure determination. Approximately 90% of the backbone resonances were assigned from a combination of triple-resonance data (HNCA, HNCO, CBCACONH, HBHACONH, HNHA and HCACO), intraresidue and sequential NOEs (3D 15N-NOESY-HMQC and 13C-HSQC-NOESY) and side-chain information obtained from the CCONH and HCCONH experiments. Preliminary analysis of the NOE pattern in the 15N-NOESY-HMQC spectrum and the 13C alpha secondary chemical shifts predicts a secondary structure for OM consisting of four alpha-helices with three intervening helical regions, consistent with the four-helix-bundle motif found for this cytokine family. As a 203-residue protein with a molecular weight of 24 kDa, Oncostatin M is the largest alpha-helical protein yet assigned.

Amino Acid Sequence↗
PubMed1996

Exploratory factor analysis of MRI brain structure measures in schizophrenia.

Much of the literature shows various regional structural brain abnormalities in schizophrenia, but the complexity and variability of brain makes it difficult to determine how these regions are related. Statistical methods which estimate factors underlying patterns of covariance have not been widely used, but could be useful for analyzing such complex data. We applied exploratory and confirmatory factor analysis procedures to specific cortical and subcortical regional brain volume measures from MRI data in 60 normal and 44 schizophrenic subjects. Basal ganglia, heteromodal cortical gray, and medial temporal lobe factors were present in both the normal and the schizophrenia groups. The factor structure observed in the normal group showed a high degree of bilateral symmetry which is present but disrupted in the schizophrenia group. In the bilateral data, the disruption is most pronounced with medial and lateral temporal lobe structures including entorhinal cortex and anterior and posterior superior temporal gyri. There was a significant correlation between the basal ganglia factor and the heteromodal cortical gray factor in the normal group that was not present in the schizophrenia group. In the unilateral data, left posterior superior temporal gyrus did not load onto any factor in the schizophrenia group. Confirmatory factor analyses showed significant differences between the two groups in factor structure. A number of specific brain regions are affected in schizophrenia, and structural relationships between groups of regions also are abnormal. The results suggest that heteromodal dorsolateral prefrontal and superior temporal cortical gray regions are structurally related, whereas inferior parietal cortical gray is less so. These results should be viewed as preliminary as the ratio of parameters to subjects was relatively low, and replication is needed. However, the results demonstrate the potential utility of latent structure methods such as factor analysis in study of complex relationships in neuropsychiatric data.

Adult↗
PubMed1995

1H, 15N, 13C and 13CO assignments and secondary structure determination of basic fibroblast growth factor using 3D heteronuclear NMR spectroscopy.

The assignments of the 1H, 15N, 13CO and 13C resonances of recombinant human basic fibroblast growth factor (FGF-2), a protein comprising of 154 residues and with a molecular mass of 17.2 kDa, is presented based on a series of three-dimensional triple-resonance heteronuclear NMR experiments. These studies employ uniformly labeled 15N- and 15N-/13C-labeled FGF-2 with an isotope incorporation > 95% for the protein expressed in E. coli. The sequence-specific backbone assignments were based primarily on the interresidue correlation of C alpha, C beta and H alpha to the backbone amide 1H and 15N of the next residue in the CBCA(CO)NH and HBHA(CO)NH experiments and the intraresidue correlation of C alpha, C beta and H alpha to the backbone amide 1H and 15N in the CBCANH and HNHA experiments. In addition, C alpha and C beta chemical shift assignments were used to determine amino acid types. Sequential assignments were verified from carbonyl correlations observed in the HNCO and HCACO experiments and C alpha correlations from the HNCA experiment. Aliphatic side-chain spin systems were assigned primarily from H(CCO)NH and C(CO)NH experiments that correlate all the aliphatic 1H and 13C resonances of a given residue with the amide resonance of the next residue. Additional side-chain assignments were made from HCCH-COSY and HCCH-TOCSY experiments. The secondary structure of FGF-2 is based on NOE data involving the NH, H alpha and H beta protons as well as 3JHNH alpha coupling constants, amide exchange and 13C alpha and 13C beta secondary chemical shifts. It is shown that FGF-2 consists of 11 well-defined antiparallel beta-sheets (residues 30-34, 39-44, 48-53, 62-67, 71-76, 81-85, 91-94, 103-108, 113-118, 123-125 and 148-152) and a helix-like structure (residues 131-136), which are connected primarily by tight turns. This structure differs from the refined X-ray crystal structures of FGF-2, where residues 131-136 were defined as beta-strand XI. The discovery of the helix-like region in the primary heparin-binding site (residues 128-138) instead of the beta-strand conformation described in the X-ray structures may have important implications in understanding the nature of heparin-FGF-2 interactions. In addition, two distinct conformations exist in solution for the N-terminal residues 9-28. This is consistent with the X-ray structures of FGF-2, where the first 17-19 residues were ill defined.

Escherichia coli↗
PubMed1994

[3H]PtdIns hydrolysis in postmortem human brain membranes is mediated by the G-proteins Gq/11 and phospholipase C-beta.

A method utilizing exogenously added [3H]PtdIns incubated with membranes prepared from postmoretem human brain has been shown to provide a means of measuring agonist-induced, guanosine 5'-O-(thiotriphosphate) (GTP[S])-dependent hydrolysis of [3H]PtdIns, thus allowing investigations of the activity of the phosphoinositide second-messenger system in accessible human brain tissue. Agonists inducing [3H]PtdIns hydrolysis include carbachol, trans-1-aminocyclopentyl-1,3-dicarboxylate (ACPD; a glutamatergic metabotropic receptor agonist), serotonin and ATP, with the latter two agonists producing the largest responses. In addition to ATP, [3H]PtdIns hydrolysis was induced by ADP and by 2-methylthio-ATP, indicating that P2-purinergic receptors mediate this process. Subtype-selective antibodies we used to identify Gq/11 and phospholipase C-beta as the G-protein and phospholipase C subtypes that mediated GTP[S]-induced and agonist-induced [3H]PtdIns hydrolysis. These results demonstrate that this method reveals that agonist-induced, GTP[S]-dependent [3H]PtdIns hydrolysis is retained in postmortem human brain membranes with properties similar to rat brain. This method should allow studies of the modulation of phosphoinositide hydrolysis in human brain and investigations of potential alterations in postmortem brain from subjects with neurological and psychiatric diseases.

Adenosine Triphosphate↗