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B D Hoit

Publications and source records attributed to B D Hoit.

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

beta2-adrenergic receptor polymorphisms at amino acid 16 differentially influence agonist-stimulated blood pressure and peripheral blood flow in normal individuals.

BACKGROUND: The Gly16 beta(2)-adrenergic receptor (beta(2)AR) polymorphism is a common variant of the beta(2)AR that displays depressed function caused by enhanced receptor downregulation in vitro compared with the Arg16 receptor. METHODS AND RESULTS: We studied 20 healthy, normotensive, nonsmoking white individuals who were homozygous for either the Arg16 (n = 10) or the Gly16 (n = 10) genotype. Plethysmographic lower-limb blood flow, blood pressure, and 2-dimensional echocardiograms were recorded at baseline and after 15-minute incremental infusions of terbutaline (100 to 300 ng/kg per minute). Baseline heart rates, blood pressures, and flows were similar in both groups, but at the maximum dose of terbutaline, limb blood flow was less (P <.05), calculated vascular resistance was greater (P <.05), and systolic and diastolic blood pressures were greater in patients with Gly16 than in those with Arg16 (both P <.05). In contrast, terbutaline-stimulated heart rates were not different. In a separate group of 20 homozygous individuals (12 Arg16, 8 Gly16), there were no differences in 2-dimensional echocardiographically determined ventricular function. CONCLUSIONS: We conclude that the Gly16 beta(2)AR polymorphism imparts attenuated vasodilatory responses to catecholamines in normal human beings and is an important genetic component in the regulation of peripheral blood flow and systemic arterial pressure.

Adrenergic beta-2 Receptor Agonists↗
PubMed2000

Alterations in Ca2+ cycling proteins and G alpha q signaling after left ventricular assist device support in failing human hearts.

OBJECTIVE: Left ventricular assist device support mechanically unloads the failing ventricle with resultant improvement in cardiac geometry and function in patients with end-stage heart failure. Activation of the G alpha q signaling pathway, including protein kinase C, appears to be involved in the progression of heart failure. Similarly down-regulation of Ca2+ cycling proteins may contribute to contractile depression in this clinical syndrome. Thus we examined whether protein kinase C activation and decreased Ca2+ cycling protein levels could be reversed by left ventricular assist device support. METHODS: Left ventricular myocardial specimens were obtained from seven patients during placement of left ventricular assist device and heart transplantation. We examined changes in protein levels of G alpha q, phospholipase C beta 1, regulators of G protein signaling (RGS), sarcoplasmic reticulum Ca2+ ATPase, phospholamban and translocation of protein kinase C isoforms (alpha, beta 1, and beta 2). RESULTS: The paired pre- and post-left ventricular assist device samples revealed that RGS2, a selective inhibitor of G alpha q, was decreased (P < 0.01), while the status of G alpha q, phospholipase C beta 1, RGS3 and RGS4 were unchanged after left ventricular assist device implantation. Translocation of protein kinase C isoforms remained unchanged. Left ventricular assist device support increased sarcoplasmic reticulum Ca2+ ATPase protein level (P < 0.01), while phospholamban abundance was unchanged. CONCLUSIONS: We conclude that altered protein expression and stoichiometry of the major cardiomyocyte Ca2+ cycling proteins rather than reduced phospholipase C beta 1 activation may contribute to improved mechanical function produced by left ventricular assist device support in human heart failure.

Adolescent↗
PubMed2000

Influence of sarcoplasmic reticulum calcium loading on mechanical and relaxation restitution.

Mechanical and relaxation restitution represent the restoration of contractile force and relaxation, respectively, in premature beats having progressively longer extrasystolic intervals (ESI); these phenomena are related to intracellular activator Ca(2+) by poorly defined mechanisms. We tested the hypothesis that the level of phospholamban [which modulates the affinity of the sarcoplasmic reticulum (SR) Ca(2+)-ATPase for Ca(2+), and thus the SR Ca(2+) load] may be an important determinant of both mechanical and relaxation restitution. Five mice with ablation of the phospholamban (PLB) gene (PLBKO), eight isogenic wild-type controls (129SvJ), eleven mice with PLB overexpression (PLBOE), and nine isogenic wild-type (FVB/N) controls were anesthetized and instrumented with a 1.4-Fr Millar catheter in the left ventricle and a 1-Fr pacemaker in the right atrium. At a cycle length of 200 ms, extrastimuli with increasing ESI were introduced, and the peak rates of left ventricular isovolumic contraction (+/-dP/dt(max)) were normalized and fit to monoexponential equations. In a subset, the protocols were repeated after ryanodine (4 ng/g) was administered to deplete SR Ca(2+) stores. The time constant of mechanical restitution in PLBKO was significantly shorter [6.3 +/- 1.2 (SE) vs. 47.7 +/- 7.6 ms] and began earlier (50 +/- 10 vs. 70 +/- 19 ms) than in 129SvJ. In contrast, the time constant of mechnical restitution was significantly longer (80.3 +/- 7.6 vs. 54.1 +/- 9.2 ms) in PLBOE than in FVB/N. The time constant of relaxation restitution was less in PLBKO than in 129SvJ (26.2 +/- 9.9 vs. 44.6 +/- 3.3, P < 0.05) but was similar in PLBOE and FVB/N (21.1 +/- 6.3 vs. 20.5 +/- 5.7 ms). Intravenous ryanodine decreased significantly the time constants of mechanical restitution in PLBOE, 129SvJ, and FVB/N but was lethal in PLBKO. In contrast, ryanodine increased the time constant of relaxation restitution. Thus 1) the phospholamban level is a critical determinant of mechanical restitution and (to a lesser extent) relaxation restitution in these transgenic models, and 2) ryanodine differentially affects mechanical and relaxation restitution. Furthermore, our data suggest a dissociation of processes within the SR that govern contraction and relaxation.

Animals↗
PubMed1999

Influence of transgenic overexpression of phospholamban on postextrasystolic potentiation.

Twelve mice with PLB overexpression (PLBOE), and 11 isogenic FVB/N wild-type (WT) controls, were anesthetized and instrumented with a 1.4 F Millar catheter in the LV and a 1 F pacemaker in the right atrium. At a cycle length of 200 ms and a fixed extrastimulus of 120 ms, extrastimuli with increasing intervals (PESI) up to 1000 ms were introduced, and the peak rates of LV isovolumic contraction (+/- dP/dtmax) were normalized and fit to monoexponential equations. In a subset of animals, the protocols were repeated after ryanodine (4 ng/g) was given to deplete SR Ca2+ stores. The time constant and the plateau of the exponential curve fits were significantly greater in PLBOE than WT (107.8 +/- 7.0 v 75.2 +/- 5.5 ms and 1.39 +/- 0.03 v 1.08 +/- 0.02, both P < 0.05). At 200, 600 and 1000 ms, the normalized dP/dt was significantly greater in PLBOE than WT. After ryanodine, normalized dP/dt was significantly decreased in PLBOE, but unchanged in WT. The protein levels of the sodium-calcium exchanger normalized to calsequestrin were increased 3.7 +/- 0.3-fold in PLBOE compared to controls. In conclusion, the phospholamban level is a critical determinant of postextrasystolic potentiation in this transgenic model, and is differentially impaired by ryanodine at long diastolic intervals in PLBOE v controls. These differences may be due in part to changes in the protein level and resultant activity of the sodium calcium exchanger.

Adenosine Triphosphatases↗
PubMed1999

Effect of congestive heart failure on in vivo canine aortic elastic properties.

OBJECTIVES: The aim of this study was to characterize fully in vivo aortic compliance over a wide range of passive distending pressures, and to study pharmacologically induced alterations in compliance using an intravascular ultrasound-based technique in the canine model of heart failure. BACKGROUND: Altered aortic compliance may influence considerably the function of the failing heart. Although some studies demonstrate that patients with heart failure have decreased aortic compliance, data from other studies are conflicting. METHODS: Aortic pressures and dimensions in seven dogs were determined both before and after pacing-induced congestive heart failure (CHF) using simultaneous micromanometer and intravascular ultrasound transducers. Decreases in aortic pressure were produced at baseline and after nitroprusside and dobutamine infusions. Inner and outer aortic circumferences were drawn at the lumen-intimal and media-adventitial borders. RESULTS: Aortic pressure-dimension (chamber) stiffness constants were greater after heart failure was produced (10.0+/-1.5 vs. 6.7+/-1.5, p < 0.05), but stress-strain stiffness (material) constants were similar (11.4+/-1.8 vs. 11.3+/-1.0, p=NS). Equivasodilating doses of nitroprusside and 10 microg/kg/min dobutamine decreased pressure-dimension stiffness constants after pacing-induced heart failure but not beforehand. The aortic wall thickness to diameter ratio was significantly greater in CHF than in the control condition (0.30+/-0.08 vs. 0.16+/-0.03, p < 0.01). CONCLUSIONS: Aortic compliance is decreased in this model of CHF, and this change is attributable primarily to vessel geometry rather than material properties. Equivasodilating doses of nitroprusside and equivalent doses of dobutamine increase aortic chamber compliance in dogs with CHF, but not in normal dogs. These data suggest that the beneficial effects of nitroprusside and dobutamine in CHF occur in part from improvement in aortic compliance.

Animals↗
PubMed1999

Noninvasive evaluation of cardiac dysfunction by echocardiography in streptozotocin-induced diabetic rats.

BACKGROUND: There has not been a noninvasive in vivo longitudinal evaluation of cardiac function in diabetic rats. The objective of this study is to examine the time course of development of cardiac dysfunction in streptozotocin (STZ)-induced diabetic rats. METHODS AND RESULTS: Cardiac function was evaluated by M-mode and Doppler echocardiography in anesthetized Wistar rats at 2, 4, 5, 6, and 8 weeks after injection with 65 mg of STZ/kg and in age-matched control rats before and after the administration of isoproterenol. Body weight (BW) was significantly less and blood glucose level significantly greater in diabetic rats compared with controls at 2 weeks and remained at these levels at all time points. The calculated left ventricular (LV) mass appeared slightly decreased in diabetic rats. However, LV mass-BW ratios were similar in controls and diabetic rats at 2, 4, and 5 weeks, but were significantly greater in diabetic rats at 6 and 8 weeks. Basal heart rate (HR) was significantly lower in diabetic rats at all time points studied. Basal LV systolic and diastolic dimensions, fractional shortening (FS), velocity of circumferential shortening (Vcf), peak emptying rate (PER), peak filling rate (PFR), and aortic peak velocity (APV) were not significantly different between controls and diabetic rats at 2 and 4 weeks. PER and PFR were significantly less in 5-week diabetic rats. However, Vcf, PER, and PFR were significantly less and FS and APV were similar at 6 and 8 weeks. Administration of isoproterenol increased HR, Vcf, FS, PFR, and PER in controls at all time points, but the increases in diabetic rats at 5, 6, and 8 weeks were less compared with those in controls. The increase in APV was significantly less in diabetic rats at all time points studied. CONCLUSION: STZ-induced diabetic rats showed bradycardia before contractile dysfunction. Overt and covert contractile dysfunction unmasked by isoproterenol begins at 5 weeks of diabetes. The overt LV systolic and diastolic dysfunction are fully manifested after 6 weeks of diabetes.

Animals↗
PubMed1999

Modulation of force-frequency relation by phospholamban in genetically engineered mice.

Phospholamban levels regulate cardiac sarcoplasmic reticulum Ca2+ pump activity and myocardial contractility. To determine whether and to what extent phospholamban modulates the force-frequency relation and ventricular relaxation in vivo, we studied transgenic mice overexpressing phospholamban (PLBOE), gene-targeted mice without phospholamban (PLBKO), and isogenic wild-type controls. Contractility was assessed by the peak rate of left ventricular (LV) isovolumic contraction (+dP/dtmax), and diastolic function was assessed by both the peak rate (-dP/dtmax) and the time constant (tau) of isovolumic LV relaxation, using a high-fidelity LV catheter. Incremental atrial pacing was used to generate heart rate vs. -dP/dtmax (force-frequency) relations. Biphasic force-frequency relations were produced in all animals, and the critical heart rate (HRcrit) was taken as the heart rate at which dP/dtmax was maximal. The average LV +dP/dtmax increased in both PLBKO and PLBOE compared with their isogenic controls (both P < 0.05). The HRcrit for LV +dP/dtmax was significantly higher in PLBKO (427 +/- 20 beats/min) compared with controls (360 +/- 18 beats/min), whereas the HRcrit in PLBOE (340 +/- 30 beats/min) was significantly lower compared with that in isogenic controls (440 +/- 25 beats/min). The intrinsic heart rates were significantly lower, and the HRcrit and the +/-dP/dtmax at HRcrit were significantly greater in FVB/N than in SvJ control mice. We conclude that 1) the level of phospholamban is a critical negative determinant of the force-frequency relation and myocardial contractility in vivo, and 2) contractile parameters may differ significantly between strains of normal mice.

Animals↗
PubMed1998

Pentameric assembly of phospholamban facilitates inhibition of cardiac function in vivo.

Phospholamban has been proposed to coexist as pentamers and monomers in native sarcoplasmic reticulum membranes. To determine its functional unit in vivo, we reintroduced wild-type (pentameric) or monomeric mutant (C41F) phospholamban in the hearts of phospholamban knockout mice. Transgenic lines, expressing similar levels of mutant or wild-type phospholamban, were identified, and their cardiac phenotypes were characterized in parallel. Sarcoplasmic reticulum Ca2+ transport assays indicated similar decreases in SERCA2 Ca2+ affinity by mutant or wild-type phospholamban. However, the time constants of relaxation and Ca2+ transient decline in isolated cardiomyocytes were diminished to a greater extent by wild-type than mutant phospholamban, even without significant differences in the amplitudes of myocyte contraction and Ca2+ transients between the two groups. Langendorff perfusion also indicated that mutant phospholamban was not capable of depressing the enhanced relaxation parameters of the phospholamban knockout hearts to the same extent as wild-type phospholamban. Moreover, in vivo assessment of mouse hemodynamics revealed a greater depression of cardiac function in wild-type than mutant phospholamban hearts. Thus, the mutant or monomeric form of phospholamban was not as effective in slowing Ca2+ decline or relaxation in cardiomyocytes, hearts, or intact animals as wild-type or pentameric phospholamban. These findings suggest that pentameric assembly of phospholamban is necessary for optimal regulation of myocardial contractility in vivo.

Animals↗
PubMed1998

Cardiac-specific overexpression of mouse cardiac calsequestrin is associated with depressed cardiovascular function and hypertrophy in transgenic mice.

Calsequestrin is a high capacity Ca2+-binding protein in the sarcoplasmic reticulum (SR) lumen. To elucidate the functional role of calsequestrin in vivo, transgenic mice were generated that overexpressed mouse cardiac calsequestrin in the heart. Overexpression (20-fold) of calsequestrin was associated with cardiac hypertrophy and induction of a fetal gene expression program. Isolated transgenic cardiomyocytes exhibited diminished shortening fraction (46%), shortening rate (60%), and relengthening rate (60%). The Ca2+ transient amplitude was also depressed (45%), although the SR Ca2+ storage capacity was augmented, as suggested by caffeine application studies. These alterations were associated with a decrease in L-type Ca2+ current density and prolongation of this channel's inactivation kinetics without changes in Na+-Ca2+ exchanger current density. Furthermore, there were increases in protein levels of SR Ca2+-ATPase, phospholamban, and calreticulin and decreases in FKBP12, without alterations in ryanodine receptor, junctin, and triadin levels in transgenic hearts. Left ventricular function analysis in Langendorff perfused hearts and closed-chest anesthetized mice also indicated depressed rates of contraction and relaxation of transgenic hearts. These findings suggest that calsequestrin overexpression is associated with increases in SR Ca2+ capacity, but decreases in Ca2+-induced SR Ca2+ release, leading to depressed contractility in the mammalian heart.

Amino Acid Sequence↗
PubMed1998

The Ile164 beta2-adrenergic receptor polymorphism adversely affects the outcome of congestive heart failure.

The beta2-adrenergic receptor (beta2AR), an important modulator of cardiac inotropy and chronotropy, has significant genetic heterogeneity in the population. Because dysfunctional betaARs play a role in the pathogenesis of the failing ventricle, we tested the hypothesis that beta2AR polymorphisms alter the outcome of congestive heart failure. 259 patients with NYHA functional class II-IV heart failure due to ischemic or dilated cardiomyopathy were genotyped and prospectively followed, with the endpoint defined as death or cardiac transplantation. The allele frequencies between this group and those of 212 healthy controls also were compared and did not differ between the groups. However, those with the Ile164 polymorphism displayed a striking difference in survival with a relative risk of death or cardiac transplant of 4.81 (P < 0.001) compared with those with the wild-type Thr at this position. Age, race, gender, functional class, etiology, ejection fraction, and medication use did not differ between these individuals and those with the wild-type beta2AR, and thus the beta2AR genotype at position 164 was the only clear distinguishing feature between the two groups. The 1-yr survival for Ile164 patients was 42% compared with 76% for patients harboring wild-type beta2AR. In contrast, polymorphisms at amino acid positions 16 (Arg or Gly) or 27 (Gln or Glu), which also alter receptor phenotype, did not appear to have an influence on the course of heart failure. Taken together with cell-based and transgenic mouse results, this study establishes a paradigm whereby genetic variants of key signaling elements can have pathophysiologic consequences within the context of a disease. Furthermore, patients with the Ile164 polymorphism and heart failure may be candidates for earlier aggressive intervention or cardiac transplantation.

Adult↗
PubMed1998

Decompensation of pressure-overload hypertrophy in G alpha q-overexpressing mice.

BACKGROUND: Receptor-mediated activation of myocardial Gq signaling is postulated as a biochemical mechanism transducing pressure-overload hypertrophy. The specific effects of Gq activation on the functional and morphological adaptations to pressure overload are not known. METHODS AND RESULTS: To determine the effects of intrinsic myocyte G alpha q signaling on the left ventricular hypertrophic response to experimental pressure overload, transgenic mice overexpressing G alpha q specifically in the heart (G alpha q-25) and nontransgenic siblings underwent microsurgical creation of transverse aortic coarctation and the morphometric, functional, and molecular characteristics of these pressure-overloaded hearts were compared at increasing times after surgery. Before aortic banding, isolated G alpha q-25 ventricular myocytes exhibited contractile depression (depressed +dl/dt and -dl/dt) and G alpha q-25 hearts showed a pattern of fetal gene expression similar to the known characteristics of nontransgenic pressure-overloaded mice. Three weeks after transverse aortic banding, G alpha q-25 left ventricles hypertrophied to a similar extent (approximately 30% increase) as nontransgenic mice. However, whereas nontransgenic mice exhibited concentric left ventricular remodeling with maintained ejection performance (compensated hypertrophy), G alpha q-25 left ventricles developed eccentric hypertrophy and ejection performance deteriorated, ultimately resulting in left heart failure (decompensated hypertrophy). The signature hypertrophy-associated progress of fetal cardiac gene expression observed at baseline in G alpha q-25 developed after aortic banding of nontransgenic mice but did not significantly change in aortic-banded G alpha q-25 mice. CONCLUSIONS: Intrinsic cardiac myocyte G alpha q activation stimulates fetal gene expression and depresses cardiac myocyte contractility. Superimposition of the hemodynamic stress of pressure overload on G alpha q overexpression stimulates a maladaptive form of eccentric hypertrophy that leads to rapid functional decompensation. Therefore G alpha q-stimulated cardiac hypertrophy is functionally deleterious and compromises the ability of the heart to adapt to increased mechanical load. This finding supports a reevaluation of accepted concepts regarding the mechanisms for compensation and decompensation in pressure-overload hypertrophy.

Animals↗
PubMed1998

Pulmonary venous flow Doppler velocities in children.

We report the results of a prospective study of pulmonary venous (PV) flow Doppler velocities in 68 normal children. We sought to establish the normal PV flow velocities in the broad pediatric population, compare these velocities to heart rate and age. In normal children, there is a wide range of PV flow velocities, most of which correlate with age and heart rate. However, the PV flow velocities and their durations do not distinguish the age groups. The peak velocity of systolic forward flow and atrial reversal flow in the pulmonary vein were independent of heart rate. The PV flow peak systolic velocity showed a weak correlation with the velocity of the mitral inflow early wave. However, the velocity of atrial reversal flow showed no correlation with the mitral inflow A velocity (late wave), which is generated by the same force of atrial contraction.

Adolescent↗
PubMed1998

Left atrial systolic and diastolic function accompanying chronic rapid pacing-induced atrial failure.

The objective of this study was to examine the hypothesis that long-term, rapid atrial pacing produces a model of atrial systolic and diastolic dysfunction but does not alter ventricular function. Eight dogs were atrially paced at 400 beats/min (3:1-5:1 ventricular response) for 6 wk and subsequently instrumented with left atrial (LA) and left ventricular (LV) sonomicrometers and micromanometers. Data were compared with those from six sham-operated controls at matched heart rates and mean LA pressures of 10 mmHg. Dogs with rapid pacing had slightly greater LA volume (10.3 +/- 4.0 vs. 7.9 +/- 4.4 ml) and reduced ejection fraction (2.2 +/- 1.4 vs. 13.0 +/- 4.0, P < 0.05), systolic ejection rate (0.3 +/- 0.1 vs. 2.8 +/- 1.2 vol/s, P < 0.05), and reservoir fraction (0.07 +/- 0.04 vs. 0.35 +/- 0.06, P < 0.05) compared with controls. LA diastolic chamber stiffness was greater after rapid atrial pacing than before (stiffness constant kc, 5.7 +/- 2.3 vs. 3.4 +/- 0.6, P < 0.05), and the ratio of transesophageal echo-determined pulmonary venous systolic to diastolic integrated flow (a measure of relative reservoir to conduit function of the LA) was less in rapidly paced dogs compared with control dogs (0.41 +/- 0.19 vs. 0.68 +/- 0.23, P < 0.05). In contrast, rapid atrial pacing did not influence LV systolic performance or lusitropy, because the LV pressure time derivative and the time constant of LV relaxation were similar in both groups. In this model of isolated atrial myopathy, increased atrial stiffness and enhanced conduit function compensate for impaired atrial booster pump and reservoir functions.

Animals↗
PubMed1997

Targeted overexpression of protein kinase C beta2 isoform in myocardium causes cardiomyopathy.

Increased cardiovascular mortality occurs in diabetic patients with or without coronary artery disease and is attributed to the presence of diabetic cardiomyopathy. One potential mechanism is hyperglycemia that has been reported to activate protein kinase C (PKC), preferentially the beta isoform, which has been associated with the development of micro- and macrovascular pathologies in diabetes mellitus. To establish that the activation of the PKCbeta isoform can cause cardiac dysfunctions, we have established lines of transgenic mice with the specific overexpression of PKCbeta2 isoform in the myocardium. These mice overexpressed the PKCbeta2 isoform transgene by 2- to 10-fold as measured by mRNA, and proteins exhibited left ventricular hypertrophy, cardiac myocyte necrosis, multifocal fibrosis, and decreased left ventricular performance without vascular lesions. The severity of the phenotypes exhibited gene dose-dependence. Up-regulation of mRNAs for fetal type myosin heavy chain, atrial natriuretic factor, c-fos, transforming growth factor, and collagens was also observed. Moreover, treatment with a PKCbeta-specific inhibitor resulted in functional and histological improvement. These findings have firmly established that the activation of the PKCbeta2 isoform can cause specific cardiac cellular and functional changes leading to cardiomyopathy of diabetic or nondiabetic etiology.

Animals↗
PubMed1997

Effects of thyroid hormone on cardiac beta-adrenergic responsiveness in conscious baboons.

BACKGROUND: Many of the cardiovascular manifestations of thyroid hormone excess resemble those produced by sympathoadrenal stimulation. The objective of this study was to determine the effects of thyroid hormone excess on myocardial beta-adrenergic expression and responsiveness to infused agonists in the primate heart. METHODS AND RESULTS: The responses of left ventricular isovolumic contraction (dP/dt(max)) and relaxation (tau) during graded dobutamine infusion were studied both before and after 4 weeks of thyroid hormone administration in 8 chronically instrumented baboons. At matched (atrially paced) heart rates, thyroid hormone significantly increased resting dP/dt(max) (3073+/-1034 versus 2318+/-829 mm Hg/s, P<.05) and decreased tau (24.0+/-5.5 versus 28.2+/-5.4 ms, P<.05). The change from baseline for dP/dt(max) and tau in response to beta1-adrenergic stimulation was significant at each dobutamine dose (2.5 to 10 microg x kg(-1) x min(-1)), but when expressed as a percent change, it was similar before versus after thyroid hormone. Similar changes were found when beta2-adrenergic stimulation was produced by terbutaline infusion in three additional baboons. beta-Adrenergic receptor (betaAR) expression was higher in five thyroxine-treated than in five control baboons (37.4+/-1.2 versus 15.7+/-3.2 fmol/mg, P<.001), and this was due to a greater increase in the beta2AR (5.9+/-1.5 to 20.6+/-1.2 fmol/mg, P<.001) than the beta1AR (9.7+/-1.7 to 16.8+/-0.1 fmol/mg, P<.01) subtype. CONCLUSIONS: In the primate heart, thyroid hormone produces positive inotropic and lusitropic effects in the resting state and upregulates both beta1AR and beta2AR, with the beta2AR increase predominating. At equivalent rates, however, thyroid hormone excess does not appear to enhance the sensitivity of left ventricular contractility and relaxation to either beta1- or beta2-adrenergic stimulation.

Adrenergic beta-Agonists↗
PubMed1997

Selective reporter expression in mast cells using a chymase promoter.

Primate alpha-chymases are mast cell neutral proteases that are involved in regulating several regulatory peptides including angiotensin II. Because of significant substrate specificity differences among the chymase group of enzymes, animal models that overexpress primate chymases are crucial for delineating the in vivo function of these enzymes. Activation of alpha-prochymase requires processing enzymes and proteoglycans found in mast cell secretory granules. Thus, the development of models overexpressing active primate chymase requires a mast cell-specific promoter. We show that the 571-base pair (bp) 5'-upstream sequence of the baboon chymase gene, which encodes an alpha-chymase, coupled to the prokaryotic lacZ gene allows the targeting of beta-galactosidase to mast cells in transgenic mice. Tissue expression of the transgene is similar to the expression of the endogenous mouse alpha-chymase mouse mast cell protease-5. A mouse mast cell line that endogenously expresses mouse mast cell protease-5 (JKras mast cells) also selectively supports the expression of this transgene. In vitro transcription studies in JKras mast cells shows the critical role of a GATA cis-regulatory motif in baboon chymase promoter, located approximately 430-bp upstream of the transcription start site. These results suggest that the 571-bp domain of the baboon chymase promoter contains most, if not all, of the mast cell-specific region of the promoter. We describe here for the first time a promoter that directs expression of transgenes specifically to mouse mast cells. This promoter should be generally applicable for dominant expression of mast cell regulatory proteins.

3T3 Cells↗
PubMed1997

Global and regional atrial function after rapid atrial pacing: an echo Doppler study.

We examined diastolic transmitral, left atrial appendage, and pulmonary venous flow pulsed-wave Doppler velocities in five dogs sedated with Valium and ketamine both before and 1 week after rapid (400 beats/min) atrial pacing with transesophageal echocardiography. Left ventricular systolic function was assessed with long-axis fractional area shortening with the use of transesophageal echocardiography. Transmitral and left atrial appendage atrial systolic and pulmonary vein systolic velocities decreased significantly. The transmitral and left atrial appendage early-to-late velocity ratios increased significantly and the pulmonary venous systolic-to-diastolic velocity ratio decreased significantly after rapid atrial pacing. Thus, 1 week of rapid atrial pacing results in impaired global and regional atrial pump function and enhanced conduit function of the left atrium with preservation of left ventricular systolic function. These findings may have implications for anticoagulation therapy in patients with atrial arrhythmia.

Animals↗