Biomedical subjects

J M Stewart

Publications and source records attributed to J M Stewart.

At least 109 records · Page 6Linked to original sources

PubMed1993

Programs for phasing by entropy maximization as implemented in Xtal3.2: a crystallographic software system.

Xtal3.2, a crystallographic software package, is an international development project involving about 40 researchers over a full spectrum of crystallographic interests. This development has been supported by many national and international agencies and commercial institutions since the first version in 1983. The 1992 release, Xtal3.2, contains software for 95 different calculations. These range from the processing of raw diffraction data to interactive molecular graphics, atomic charge estimation, electronic publication preparation, and the structure solution and refinement of small and large molecules. Tests of the Xtal programs for phase determination and phase refinement by the application of 'maximum entropy' are presented.

Journal Article↗

PubMed1993

A proton magnetic resonance study of two synthetic agonist-antagonist pairs of bradykinin analogues.

The conformation of two agonist-antagonist pairs of bradykinin (Arg1-Pro2-Pro2-Gly4-Phe5-Ser6-Pro7-Phe8-Arg9) analogues were studied in CD3OH/H2O solution by 1H-nmr techniques. The first agonist peptide studied, D-Arg0-Arg1-Pro2-Hyp3-Gly4-Thi5-Ser6-Pro7- Thi8-Arg9, differs from the bradykinin sequence by the addition of D-Arg0, the replacement of the Phe moieties in positions 5 and 8 by Thi (Thi = beta-(2-thienyl)-L-alanine), and Hyp3 (Hyp = L-4-hydroxy-L-proline) in position 3. In the corresponding antagonist sequence, Pro7 is replaced by D-Phe7. The second agonist-antagonist pair studied does not contain the D-Arg0 residue, which is present only to slow down the rate of metabolism. Based on complete resonance assignments from two-dimensional total correlation spectroscopy and rotating frame nuclear Overhauser effect spectroscopy spectra at 500 MHz, the peptides were analyzed in terms of intraresidue, sequential, and medium-range nuclear Overhauser effects, amide proton temperature coefficients, and vicinal coupling constants. Both agonist peptides show clear evidence for the existence of a type I beta-turn comprising the C-terminal residues Ser6-Pro7-Thi8-Arg9 in fast conformational equilibrium with extended structures throughout. Although the conformational space is dominated by extended structures, the presence of the beta-turn is spectroscopically clearly discernible. The two antagonist peptides, on the other hand, do not show evidence of turn formation but rather the presence of an extended conformation with some irregularities in the N-terminal region of the peptide. While the existence of a turn at the C-terminal end of bradykinin and its analogues with agonist activity has been predicted by empirical calculations and measurements in very apolar solvents, this study, for the first time, provides evidence based on physical data in a polar solvent environment that the turn is present, that it is type I and that it is essential for agonist activity. In the particular solvent used in these studies, the Pro7 to D-Phe7 substitution precluded the formation of the turn for the C-terminal residues of the antagonist.

Amino Acid Sequence↗

PubMed1993

Proton magnetic resonance studies of bradykinin antagonists.

Bradykinin (BK) is a peptide hormone with sequence Arg1-Pro2-Pro3-Gly4-Phe5-Ser6-Pro7-Phe8-Arg9 and has been implicated in a multitude of pathophysiological processes such as the ability to lower systemic blood pressure and stimulate pain. BK analogues having bulky, beta-branched D-aliphatic residues at position 7 combined with bulky L-aliphatic residues at position 8 have now been observed to be strong antagonists. Conformational studies based on two-dimensional nmr experiments in methanol/water (80/20 v/v) were carried out on several such active antagonists in a polar solvent. Included in this study were the very active antagonists, [D-Arg0,Hyp3,Thi5,D-Cpg7,Cpg8]-BK [Cpg: alpha-cyclo-pentyl-glycine; Hyp: trans-4-hydroxy-L-proline; Thi: beta-(2-thienyl)-L-alanine] (I), [D-Arg0,Hyp3,D-Cpg7,Cpg8]-BK (II), as well as its variant with D-Cpg7 replaced by Cpg7, namely [D-Arg0,Hyp3,Cpg7,Cpg8]-BK (III). A turn-like structure, which coexists with the extended conformation, was observed between residues 2 and 5 for the most active antagonists I and II, in direct correlation with the peptide activities. No turn-like structure was found for residues 6-9. In peptide III, a turn-like structure was not identified. The existence of a turn at the C-terminal end of bradykinin and its analogues has been predicted by empirical calculations and supported by nmr measurements. But the present nmr study on the most active antagonists (I, II) does not support this hypothesis. Instead, the data suggest that a turn-like structure between residues 2 and 5 could be important for antagonist activity. Finally, one weak inhibitor [D-Cpg7]-BK (IV) showed no defined secondary structure.

Amino Acid Sequence↗

PubMed1993

The kinin system in inflammation.

Bradykinin and related kinins are important mediators of most kinds of inflammation. Mechanisms of kinin production in injury, infection and immune system activation are discussed. Bradykinin antagonists show great promise for development of new drugs for inflammation.

Amino Acid Sequence↗

PubMed1993

Neuropeptide processing in pathophysiology.

Neuropeptides (peptides synthesized and secreted by neurons) perform many functions in the central nervous system as neurotransmitters, neuromodulators and neurotrophic factors. Neuropeptides are important regulators of amine neurotransmitter release, and can be identified as playing important roles in several pathological states. Neuropeptides are synthesized as protein precursors that are processed enzymatically to yield the biologically active peptides. Many neuropeptides having defined roles are further processed enzymatically to yield fragments having totally different actions. Examples discussed are substance P, adrenocorticotrophic hormone, endorphins and vasopressin.

Amino Acid Sequence↗

PubMed1993

The aggregation properties of some bradykinin analogs.

Amino Acid Sequence↗

PubMed1993

Synthesis and stability of 3-nitro-2-pyridinesulfenyl chloride (NpysCl).

3-Nitro-2-pyridinesulfenyl chloride (NpysCl) is the starting material for the synthesis of N-, O- and S-Npys-protected amino acids. Two efficient, novel synthetic routes to NpysCl are described. The stability of NpysCl was determined in a variety of solvents, with and without base, to determine the most suitable solvent and base for the synthesis of N-Npys amino acids. The syntheses of Npys-Ala and Boc-Lys(Npys) tert-butylammonium salt are also described.

Alanine↗

PubMed1993

Bradykinin-induced mesenteric vasodilation is mediated by B2-subtype receptors and nitric oxide.

We investigated mechanisms mediating bradykinin (BK)-induced anterior mesenteric artery (AMA) vasodilation in anesthetized rats. The velocity of blood flowing (VBF) in the AMA was measured with pulsed Doppler velocimetry, and arterial pressure (BP) was measured with a pressure transducer. Drugs were infused through an intra-aortic catheter positioned proximal to the AMA origin. AMA conductance (C) was calculated from mean VBF/BP and expressed as percent of control C. BK infusion (10-1,000 ng.kg-1.min-1) increased C significantly (Cmax = 201 +/- 18%, ED50 = 100 ng.kg-1.min-1, P < 0.01 for all doses). A B2-subtype receptor antagonist, D-Arg,[Hyp3,Thi5.8,D-Phe7]BK, administered at 10(5) ng.kg-1.min-1 before or during BK infusion, inhibited the vasodilation by 73 +/- 7 and 103 +/- 7%, respectively. A nitric oxide (NO) synthesis inhibitor, NG-nitro-L-arginine, administered at 5.0 mg/kg 15 min before BK, inhibited the hyperemia by 61 +/- 8%. Neither a B1-receptor antagonist nor intrajejunal capsaicin inhibited BK-induced vasodilation. BK-evoked, dose-dependent, mesenteric vasodilation in rats appears to be mediated partly by B2-receptors and endogenous NO generation.

Animals↗

PubMed1993

Role of tachycardia and V wave wall stress in the release of ANF during volume loading.

Heart rate is believed to be a primary determinant of atrial natriuretic factor (ANF) release; however, we postulated that V wave wall stress is the main signal for ANF release during volume loading. Conscious dogs were volume loaded with 1,000 ml of saline while the heart rate was allowed to vary and on another day while the heart was paced at 240 beats/min. During volume loading, plasma ANF increased from 58 +/- 10 to 221 +/- 31 pg/ml (P < 0.05), and heart rate increased from 97 +/- 3 to 182 +/- 9 beats/min. During pacing, plasma ANF increased from 42 +/- 8 to 85 +/- 10 pg/ml. A wave atrial pressures were increased during pacing and were fixed due to contraction against a closed atrioventricular valve. Volume loading increased plasma ANF to 180 +/- 25 pg/ml. V wave atrial pressure increased markedly, and V wave atrial wall stress increased from 25 +/- 10 to 131 +/- 44 and from 7 +/- 4 to 116 +/- 25 g/cm2, respectively. A wave pressure and wall stress were not increased by volume loading. V wave wall stress and ANF were significantly correlated. V wave atrial wall stress accounts for the majority of change in plasma ANF (60%), while tachycardia and A wave wall stress account for a smaller fraction (40%) of change.

Animals↗

PubMed1993

Is bradykinin (BK) a physiological vasodilator in the gut?

The physiological role of bradykinin (BK) as a mesenteric vasoregulator was explored. This nonapeptide is a potent vasodilator substance when administered exogenously in multiple in vivo models and is a smooth muscle relaxant when added to in vitro preparations. BK is naturally occurring in the gut wall. The substrate for BK, as well as the biosynthetic and metabolizing systems are present in the blood, the vascular wall, immunological cells, and perivascular neurons. BK B2 and B1 receptors have been characterized with sympathetic agonist and antagonist substances, and the receptors are present on mesenteric endothelial cells and myocytes. BK interacts with multiple endogenous mesenteric vasodilator mediators, such as nitric oxide, prostacyclin, and neuropeptides. Taken together this evidence supports the functional importance of BK as a normal vasodilator in the gut.

Amino Acid Sequence↗

PubMed1993

Congenital conductive hearing loss: the need for early identification and intervention.

Pediatricians are familiar with conductive hearing loss, but to many it is synonymous with otitis media of some type. There is a group of children who have anatomic abnormalities of the external or middle ear causing their hearing losses. The records of 565 hearing-impaired children were reviewed, and 54 with non-otitis-related conductive hearing loss were found. Date of birth, degree of loss, diagnosis or presence of any associated abnormalities, and ages of confirmation and intervention were studied. There were 22 children with microtia, 14 with abnormalities of the external auditory canal and normal pinnae, 15 with obvious dysmorphic features including 9 who had syndromes well known to be associated with hearing abnormalities, and only 3 who had isolated hearing loss. Despite all these clues, hearing loss was diagnosed in only 10 children (excluding those with bilateral microtia) before 12 months of age although their average hearing loss was 45 dB. It is important for the primary care provider to recognize these children as early as possible as their loss is permanent and almost always more severe than that seen in otitis.

Age Factors↗

PubMed1992

Kinetics of amide proton exchange in helical peptides of varying chain lengths. Interpretation by the Lifson-Roig equation.

The kinetics of amide proton exchange (1H----2H) have been measured by proton nuclear magnetic resonance spectroscopy for a set of helical peptides with the generic formula Ac-(AAKAA)m Y-NH2 and with chain lengths varying from 6 to 51 residues. The integrated intensity of the amide resonances has been measured as a function of time in 2H2O at pH* 2.50. Exchange kinetics for these peptides can be modeled by applying the Lifson-Roig treatment for the helix-to-coil transition. The Lifson-Roig equation is used to compute the probability that each residue is helical, as defined by its backbone (phi, psi) angles. A recursion formula then is used to find the probability that the backbone amide proton of each residue is hydrogen bonded. The peptide helix can be treated as a homopolymer, and direct exchange from the helix can be neglected. The expression for the exchange kinetics contains only three unknown parameters: the rate constant for exchange of a non-hydrogen-bonded (random coil) backbone amide proton and the nucleation (v2) and propagation (w) parameters of the Lifson-Roig theory. The fit of the exchange curves to these three parameters is very good, and the values for v2 and w agree with those derived from circular dichroism studies of the thermally-induced unfolding of related peptides [Scholtz, J.M., Qian, H., York, E.J., Stewart, J.M., & Baldwin, R.L. (1991) Biopolymers (in press]).

Amides↗

PubMed1992

Differential and coordinate responses of the human genes encoding the heat stable alkaline phosphatases to cAMP and sodium butyrate in the choriocarcinoma line JEG-3.

Human heat stable alkaline phosphatases are encoded by two closely related genes: the PLAP-1, which specifies the term placental enzyme, and the PLAP-2, which is expressed primarily in germ cells. In the choriocarcioma line JEG-3, 8-Br-cAMP induced the accumulation of the mRNA of both genes, while sodium butyrate induced the accumulation of PLAP-2 transcripts only. Each agent increased the transcription rate of one or both of the genes, as assayed by run-on transcription. In transfection of JEG-3 cells with PLAP promoters fused to the firefly luciferase gene, the activity of the PLAP-2 promoter (but not PLAP-1) was induced with sodium butyrate, while both promoters were induced by 8-Br-cAMP. Inducibility of the PLAP-2 promoter by 8-Br-cAMP was still observed when the promoter was shortened to -103, leaving intact a sequence resembling a cAMP response element. The extent of transcriptional activation by either agent was not sufficient to explain the accumulation of PLAP mRNA. These studies suggest that both transcriptional and posttranscriptional processes are involved in the induction of the PLAP-1 and PLAP-2 gene in JEG-3 cells.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1992

Bradykinin antagonists: the state of the art.

The availability of practical bradykinin antagonists has revolutionized research in the kinin field. Development of the first antagonists and progress to the present time are reviewed.

Amino Acid Sequence↗

PubMed1992

Pseudopeptide analogs of bradykinin and bradykinin antagonists.

Over 100 reduced-bond analogs of bradykinin and bradykinin antagonists were designed, synthesized, and assayed in the classic smooth muscle and blood pressure assays in a further study of the structure-activity relationships of bradykinin. Both potent BK-like agonists and antagonists of BK activity were found among the reduced-bond analogs.

Amino Acid Sequence↗

PubMed1992

Bradykinin antagonists do not require a D-aromatic amino acid residue at position 7.

More than 200 new analogs of bradykinin have been synthesized and assayed. Analogs containing a D-aliphatic amino acid residue at position 7 and an L-aliphatic residue at position 8 are very potent antagonists of BK action on classical systems. Thus the "conventional wisdom" that a D-aromatic amino acid residue is required at sequence position 7 of BK antagonists must be modified.

Amino Acid Sequence↗

PubMed1992

Evaluation of cutaneous algogenic responses to bradykinin and inhibition by bradykinin analogs.

Bradykinin has long been postulated to have a major role in physiologic human pain production. We have developed a method to systematically quantify the algogenic response to bradykinin in human skin by the direct application of bradykinin to suction blister bases. After determination of the pain threshold to bradykinin, we examined the ability of synthetic peptide analogs of bradykinin to block bradykinin-induced pain. One of the six analogs (B4642) inhibited bradykinin-induced pain with at least one concentration in all subjects tested. This result could not be predicted on the basis of inhibition of bradykinin binding to its guinea-pig ileum receptor or to previous studies of bradykinin-induced pain production in animal models. This peptide may have an eventual role as a physiologic pain inhibitor.

Amino Acid Sequence↗

PubMed1992

The substance P fragment SP(1-7) stimulates motor behavior and nigral dopamine release.

Earlier studies have shown that the undecapeptide substance P (SP) alters motor behavior and dopamine metabolism following injection into the substantia nigra (SN) in rat, even though the SN appears largely devoid of SP-specific (NK-1) receptors. In this report, intra-nigral injections of the amino-terminal SP fragment SP(1-7) enhanced rearing, sniffing and locomotor activity, and increased the nigral DOPAC-to-DA ratio. In addition, SP(1-7) increased 3H-DA release from the SN in vitro. These findings suggest that some of the effects of nigral SP on motor behavior and dopamine release are mediated by amino-terminal fragments of SP.

3,4-Dihydroxyphenylacetic Acid↗