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Biomedical subjects

J M Stewart

Publications and source records attributed to J M Stewart.

At least 19 recordsLinked to original sources
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↗
PubMed1992

Chronic elevation of norepinephrine in conscious dogs produces hypertrophy with no loss of LV reserve.

Elevated plasma catecholamine levels may cause both myocardial hypertrophy and tissue damage. To determine whether the left ventricle from dogs with chronic norepinephrine infusion can sustain additional functional loads, we altered ventricular preload or afterload and determined both global and left ventricular (LV) wall function. Dogs were instrumented to measure LV wall function, LV internal base diameter, and LV pressures and were allowed to fully recover. Preload was altered by volume loading and afterload by injection of phenylephrine. Osmotic infusion pumps were implanted to continuously release norepinephrine at 0.5 micrograms.kg-1.min-1 for 28 days, and the volume loading and phenylephrine were repeated on days 14 and 28. Heart rate decreased, whereas there were no differences in mean arterial pressure, maximum first derivative of LV pressure (dP/dt), LV dP/dt/developed pressure of 40 mmHg, LV dP/dt/end-diastolic circumference, slope of the pressure-diameter relation, peak systolic wall stress, LV/end-diastolic diameter, or LV/end-systolic diameter during norepinephrine infusion. Diastolic and systolic wall thickness and chamber weights were increased (P less than 0.05). Indexes of diastolic function, including end-diastolic pressure, end-diastolic pressure-end-diastolic diameter relationship, maximum negative dP/dt, and the time constant (tau) were unchanged after chronic norepinephrine infusion, although maximum end-diastolic pressure during volume loading was increased from 17.7 +/- 2.0 to 21.7 +/- 1.0 mmHg. Chronic norepinephrine infusion did not alter tau, and tau increased equivalently with phenylephrine injection in both normal (36 +/- 1 to 62 +/- 5 ms) dogs and in those chronically infused with norepinephrine (36 +/- 1 to 56 +/- 5 ms).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Bilateral atrial appendectomy abolishes increased plasma atrial natriuretic peptide release and blunts sodium and water excretion during volume loading in conscious dogs.

The atrial appendages contain most of the atrial natriuretic factor (ANF) in the mammalian heart, and atrial appendage mechanical function predicts ANF secretion during volume loading. To demonstrate the crucial role of the atrial appendages in ANF release, we first measured hemodynamics and changes in plasma ANF after injection of 1,000 ml i.v. normal saline in conscious dogs and again after bilateral atrial appendectomy; we next measured changes in renal function using infusions of atriopeptin 24 to achieve plasma levels corresponding to levels achieved during volume loading; and we lastly measured renal function during acute volume expansion and also after atrial appendectomy. Plasma ANF increased from 65 +/- 11 to 246 +/- 54 pg/ml after volume loading but did not increase after atrial appendectomy. Atrial appendectomy did not alter the tachycardia or hemodynamic effects of volume loading. Infusion of 10 ng/kg/min atriopeptin 24 increased plasma ANF from 50 +/- 9 to 234 +/- 54 pg/ml, increased urine output 34 +/- 10%, and increased sodium excretion 62 +/- 10% in dogs with intact atrial appendages. Renal function was compared in dogs before atrial appendectomy: 20, 40, and 60 minutes after volume loading, urine flow rate increased by 5.9 +/- 0.5, 6.9 +/- 0.4, and 4.4 +/- 0.8 ml/min, while sodium excretion increased by 717 +/- 60, 839 +/- 84, and 582 +/- 57 mueq/min. After atrial appendectomy urine flow rate increased 2.1 +/- 0.7, 2.7 +/- 0.7, and 2.0 +/- 0.6 ml/min, and sodium excretion increased only by 327 +/- 110, 324 +/- 77, and 340 +/- 92 mueq/min (p less than 0.01) during volume loading.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Evidence for renal kinins as mediators of amino acid-induced hyperperfusion and hyperfiltration in the rat.

This study examined the role of tissue kallikrein and kinins in renal vasodilation produced by infusion of amino acids (AA). In rats fed a 9% protein diet for 2 wk, intravenous infusion of a 10% AA solution over 60-90 min reduced total renal vascular resistance and increased glomerular filtration rate (GFR) by 25-40% and renal plasma flow (RPF) by 23-30% from baseline. This was associated with a two- to threefold increase in urinary kinin excretion rate. Acute treatment of rats with aprotinin, a kallikrein inhibitor, resulted in deposition of immunoreactive aprotinin in kallikrein-containing connecting tubule cells and inhibited renal kallikrein activity by 90%. A protinin pretreatment abolished the rise in urinary kinins and prevented significant increases in GFR and RPF in response to AA. In a second group of rats pretreated with a B2 kinin receptor antagonist, [DArg Hyp3, Thi5,8 D Phe7]bradykinin, AA infusion raised urinary kinins identically as in untreated controls, but GFR and RPF responses were absent. Aprotinin or the kinin antagonist produced no consistent change in renal function in rats that were not infused with AA.AA-induced increases in kinins were not associated with an increase in renal kallikrein activity. Notably, tissue active kallikrein level fell 50% in AA-infused rats. These studies provide evidence that kinins generated in the kidney participate in mediating renal vasodilation during acute infusion of AA.

Amino Acids↗
PubMed1992

Diversity of B2 bradykinin receptors with nanomolar affinity expressed in passaged IMR90 human lung fibroblasts.

IMR90 human fetal lung fibroblasts express bradykinin receptors activating the pathway for biosynthesis of PGE2. A receptor of the B2 subtype stimulates half-maximal PGE2 production at 4.8 nM bradykinin, and maximal output takes place at 25 nM bradykinin. Radioligand binding studies reveal a population of [3H]bradykinin binding sites whose affinity correlates with this B2 receptor's biologic activity, with a KD of 2.5 nM. As IMR90 cells reach 60% of their defined life span in culture, they spontaneously induce expression of a second site of lower affinity, with half-maximal binding of [3H]bradykinin at 44 nM. This second site displays a characteristic primary B2 receptor recognition profile, but differs from the 2.5 nM site on a secondary level in recognition among different B2 ligands. Bradykinin is the most potent ligand at both sites; they each preferentially recognize an N-terminal extended bradykinin peptide construct having selectivity for the rat myometrial B2 receptor, suggesting that both sites have structural features in common. However, they display diversity in their order of preference for Met-Lys-bradykinin versus Lys-Lys-bradykinin; at the 44 nM site this order is completely reversed from the order of potency exhibited at the 2.5 nM site. Expression of the second site changes the manner in which these fibroblasts control their PGE2 production; it affords a graded response of PGE2 production at bradykinin levels beyond those which would normally saturate the 2.5 nM site. The inducibility of the 44 nM site in cultured fibroblasts addresses in vivo conditions in an inflammatory environment where continuing generation of bradykinin-related peptides takes place and presents a possible mechanism for overriding constraints that would otherwise limit the progression of inflammation.

Amino Acid Sequence↗
PubMed1992

Influence of a kinin antagonist on acute hypotensive responses induced by bradykinin and captopril in spontaneously hypertensive rats.

We have evaluated the effects of a B2 receptor antagonist (B5630) of kinins on BK and captopril-induced acute hypotensive responses in anaesthetized SHR. Intravenous treatment of BK (1.0 microgram) and captopril (0.3 mg/kg) caused significant (p < 0.05) fall in the SBP and DBP. Whereas BK caused greater fall in the SBP (p < 0.05), DBP (p < 0.01) and duration of hypotension (p < 0.05) when administered after captopril (Fig 1 and 2). All the hypotensive effects of BK and captopril were significantly antagonised (p < 0.05) in the presence of B5630 (2.0 mg/kg). Further, the duration of hypotensive responses of BK and captopril were blocked (p < 0.05) by B5630. The agonists and BK-antagonist did not cause significant (p > 0.05) alterations in HR during the entire investigation. These findings provide evidence to support the suggestion that B2 receptor might be involved in the regulation of the hypotensive actions of BK and captopril. Kinins should also have valuable functions in the antihypertensive property of captopril-like drugs.

Animals↗
PubMed1992

Paraneoplastic syndromes for the primary care physician.

To summarize, we have reviewed the major endocrine, hematologic, neurologic, dermatologic, and miscellaneous paraneoplastic syndromes. These entities are of great importance to the primary care physician because, when recognized, they can permit early diagnosis of the neoplasm. Further, treatment can afford palliation of symptoms. We can expect more definite identification of the causes of the paraneoplastic syndromes in the future because more of the substances that are released by tumors will be identified and analyzed. We need to watch with anticipation for the development of specific treatments for the paraneoplastic syndromes as the causes are identified.

Endocrine System Diseases↗
PubMed1992

Pharmacologic myocardial perfusion stress tests. Adenosine thallium stress test.

Thallium-201 myocardial perfusion scintigraphy assists in detecting coronary artery disease and assessing the significance of known stenoses. An adenosine infusion protocol is discussed in conjunction with presentation of results obtained in a community hospital setting. Primary indications for pharmacologic stress maneuvers include: Evaluation of patients unable or unwilling to exercise to adequate levels of workload and patients undergoing preoperative evaluation (particularly in patients undergoing vascular surgery). Experience in a community hospital setting attests to the safety of this pharmacologic protocol while showing a diagnostic accuracy similar to results reported in peer review literature. The method has been readily received by referring internists and cardiologists and is associated with a 21% increase in nuclear myocardial perfusion stress tests during the first six months of 1991.

Adenosine↗
PubMed1991

Antagonist binding reveals two heterogenous B2 bradykinin receptors in rat myometrial membranes.

In rat myometrial membranes, two bradykinin binding sites with K1 values of 18 pM and 5.6 nM were identified. Three potent bradykinin antagonists were tested for their ability to compete for [3H]bradykinin binding. Two of them, D-Arg[Hyp3,Thi5.8,D-Phe7]bradykinin and [Hyp3,Thi5.8,D-Phe7]bradykinin, also bound to both the high- (KH) and the low-affinity (KL) site whereas [Thi5.8,D-Phe7]bradykinin identified only the low-affinity bradykinin receptor. There is a close correlation between the antagonistic potencies and the KL site affinities.

Animals↗
PubMed1991

Bradykinin-induced burst of prostaglandin formation in osteoblasts is mediated via B2 bradykinin receptors.

The receptor subtype mediating the bradykinin (Bk)-induced burst of prostaglandin formation in osteoblasts has been studied. Bk, but not des-Arg9-Bk, induced the formation of prostaglandin E2 and prostacyclin in neonatal mouse calvarial bones incubated for 30 minutes. Bk-induced prostaglandin synthesis is neonatal mouse calvarial bones was significantly inhibited by the B2 Bk receptor antagonist D-Arg0[Hyp3,Thi5,8,D-Phe7]-Bk. The B2 Bk receptor agonists Bk and Lys-Bk, but not the B1 Bk receptor agonist des-Arg9-Bk, caused a rapid burst (5 minutes) of prostaglandin E2 and prostacyclin formation in isolated osteoblast-like cells from neonatal mouse calvarial bones and in the murine osteoblastic cell lineage MC3T3-E1. When comparing the relative potency of different kinin analogs on their stimulatory effect on prostaglandin formation in isolated osteoblast-like cells and in MC3T3-E1 cells, we found that Bk = Lys-Bk greater than Met-lys-Bk much much greater than Des-Arg9-Bk. Bk-induced prostaglandin synthesis in isolated osteoblast-like cells and in MC3T3-E1 cells was inhibited by D-Arg0[Hyp3,Thi5,8,D-Phe7]-Bk, whereas the B1 Bk receptor antagonist des-Arg9-Leu8-Bk had no effect. Total binding of 3H-Bk (3-20 nM; 4 degrees C) to whole MC3T3-E1 cells reached a maximum after 4-5 h. An excess of nonradioactive Bk (1 microM) reduced cell-associated radioactivity by 20-30%. The B2 Bk receptor agonist Bk and the B2 Bk receptor antagonist D-Arg0[Hyp3,Thi5,8,D-Phe7]-Bk were able to reduce specific binding, but the B1 Bk receptor agonist des-Arg9-Bk was unable to reduce the specific binding of 3H-Bk to whole MC3T3-E1 cells. These findings indicate the presence of B2 Bk receptors on osteoblasts coupled to enhanced prostaglandin synthesis.

Amino Acid Sequence↗