Oscillometric 24-h ambulatory blood pressure reference values in Hong Kong Chinese children and adolescents.

OBJECTIVES: We aimed to establish community-based normal reference values of 24-h ambulatory blood pressure monitoring (ABPM) for Chinese children and adolescents. Furthermore, we investigated how excluding overweight children affects BP percentiles and compared them with German references. METHODS: In this territory-wide cross-sectional prospective cohort study, 1445 Hong Kong Chinese children and adolescents aged 8-17 years with body height between 119 and 185 cm were recruited. Their ABPM assessment was performed using validated arm oscillometric recorders (A&D TM-2430) and complied with American Heart Association's recommendations. The reference tables were constructed using the LMS method to normalize skewed distribution of ABP data to sex and age or height. RESULTS: The ambulatory BP was higher among boys and the difference between boys and girls progressively widened with age. An increasing trend in daytime and night-time SBP and DBP with age and height was observed in both sexes. The age-specific and sex-specific 95th percentiles from nonoverweight children (n=1147; 79%) were lower than the whole cohort by up to 2.5 and 1 mmHg for SBP and DBP, respectively. In comparison, our overall and nonoverweight reference standards were generally higher than corresponding German references. CONCLUSION: The study provides ambulatory BP standards for Chinese children, with sex-related age-specific and height-specific percentiles. Further longitudinal studies are required for investigating its clinical utility in Chinese.

Early time-dependent dynamic changes of TBET and GATA3 mRNA expressions in patients with acute coronary syndrome.

BACKGROUND: T-box expressed in T cells (TBET) and guanine adenine thymine adenine sequence-binding protein 3 (GATA3) play important roles in the differentiation of Th1 and Th2 subsets, which contributes to the progression of acute coronary syndrome (ACS). OBJECTIVE: This study aimed to investigate the temporal change of TBET/GATA3 mRNA ratio in ACS. METHODS: Thirty-three patients suspected of ACS with symptom onset within 24 hours were recruited. Blood samples were taken after arrival at the emergency department and at hourly intervals until the 6th hour. The mRNA expressions of TBET and GATA3 were quantified by a real-time RT-qPCR. RESULTS: The TBET/GATA3 mRNA ratio was elevated dramatically in patients with acute myocardial infarction (AMI) and exhibited biphasic M-shaped release kinetics with two distinct peaks. The ratio was elevated 2 hours after symptom onset, dropped to the lowest level at 10 hours, and rose to the second peak at 14 hours. A similar biphasic M-shaped curve was observed in AMI patients with blood samples taken prior to any intervention. CONCLUSIONS: The TBET/GATA3 mRNA ratio was elevated in AMI patients throughout most of the first 20 hours after symptom onset. The biphasic M-shaped release kinetics was more likely to reflect pathophysiological changes rather than treatment effects.

Left atrial appendage closure with AMPLATZER cardiac plug for stroke prevention in atrial fibrillation: initial Asia-Pacific experience.

BACKGROUND: Left atrial appendage (LAA) is the main source of left atrial thrombus that causes stroke in patients with non-valvular atrial fibrillation (NVAF). This study reported the initial safety, feasibility, and 1-yr clinical outcomes following AMPLATZER cardiac plug (ACP) implantation in Asia-Pacific region. METHODS: Twenty NVAF patients (16 males, age 68 ± 9 yr) with high risk for developing cardioembolic stroke (CHADS(2) score: 2.3 ± 1.3) and contraindications to warfarin received ACP implants from June 2009 to May 2010. Patients received general anesthesia (n = 9) or controlled propofol sedation (n = 11) and the procedures were guided by fluoroscopy and transesophageal echocardiography (TEE). Clinical follow-up was arranged at 1 month and then every 3 months after implantation, whereas, a TEE was scheduled at 1 month upon completion of dual anti-platelet therapy. RESULTS: The LAA was successfully occluded in 19/20 patients (95%) at two Asian centers. One procedure was abandoned because of catheter-related thrombus formation. Other complications included coronary artery air embolism (n = 1) and TEE-attributed esophageal injury (n = 1). The median procedural and fluoroscopic times were 79 (IQR: 59-100) and 18 (IQR 12-27) minutes, respectively. The mean size of implant was 23.6 ± 3.1 mm. The average hospital stay was 1.8 ± 1.1 days. Follow-up TEE showed all the LAA orifices were sealed without device-related thrombus formation. No stroke or death occurred at a mean follow-up of 12.7 ± 3.1 months. CONCLUSIONS: Our preliminary data suggested LAA closure with ACP is safe, feasible with encouraging 1-yr clinical outcomes. Further large-scaled trials are needed to confirm the efficacy of this device.

Atrial dysfunction and interatrial dyssynchrony predict atrial high rate episodes: insight into the distinct effects of right atrial appendage pacing.

INTRODUCTION: Right atrial (RA) appendage pacing may prolong atrial conduction time (ACT). This study aimed to investigate if RA appendage pacing can induce intra- and interatrial dyssynchrony and if atrial dysfunction and dyssynchrony can predict atrial high rate episodes (AHREs) in the first year after pacing. METHODS AND RESULTS: Patients implanted with dual-chamber pacemakers for symptomatic bradycardia were enrolled. Cumulative percentage of RA appendage pacing (Cum%AP) during 1-year follow-up and AHREs were recorded. Full Doppler echocardiography studies were performed before implantation and 1 year after pacing. ACT and peak atrial velocities (Sm-la, Em-la, Am-la) were measured. One hundred ten patients (age 70.5 ± 11 years; 53 males) were recruited and completed 1-year follow-up. ACT of both RA and left atrial (LA) were more prolonged in patients with Cum%AP > 75% than those with <25%. Intra- and interatrial dyssynchrony was more obvious in patients with Cum%AP > 75% (22.3 ± 12.2 milliseconds vs 9.5 ± 6.2 milliseconds; 53.9 ± 29.7 milliseconds vs 19.7 ± 17.3 milliseconds; both P < 0.001). AHREs occurred in 29% of patients. Atrial pump function and interatrial dyssynchrony independently predicted AHREs in multivariate analysis. Receiver operating characteristic curve provided a cutoff value of Am-la <5.3 cm/s, which predicted AHREs with a sensitivity of 71% and a specificity of 75% (area under the curve, 0.822; P < 0.001). CONCLUSION: RA appendage pacing causes atrial conduction delay with intra- and interatrial dyssynchrony. Atrial dysfunction and interatrial dyssynchrony are related to AHREs in the first year after pacing.

Deleterious effect of right ventricular apical pacing on left ventricular diastolic function and the impact of pre-existing diastolic disease.

AIMS: Right ventricular apex (RVA) pacing may have deleterious effects on left ventricular (LV) systolic function, but its impact on LV diastolic function has not been explored. METHODS AND RESULTS: Ninety-seven patients with sinus node dysfunction and ejection fraction (EF) ≥ 50% with permanent RVA pacing were randomly programmed to V-sense and V-pace modes and examined by echocardiography. Tissue Doppler imaging was employed to assess myocardial systolic velocity (S') and early diastolic velocity (E') at the mitral annulus. Systolic dyssynchrony was assessed using 12 LV segmental model (Ts-SD). Switching from V-sense to V-pace resulted in the worsening of both diastolic and systolic functions as shown by the decreased EF, reduced mean E' and S' velocities, as well as increase in LV volume and Ts-SD (all P< 0.001). Reduction of mean E' and S' of ≥ 1 cm/s occurred in 35 (36%) and 45 (46%) patients, respectively. In pre-defined subgroup analysis, only patients with pre-existing LV diastolic dysfunction had a significant reduction of mean E' and S' (both P< 0.001) even after age adjustment. Multivariate logistic regression analysis showed that independent factors for the reduction of mean E' ≥ 1 cm/s or mean S' ≥ 1 cm/s at V-pace were pre-existing LV diastolic dysfunction [odds ratio (OR): 4.735, P= 0.007 for E'; OR: 3.307, P= 0.022 for S'] and systolic dyssynchrony at V-pace (OR: 5.459, P= 0.007 for E'; OR: 2.725, P= 0.035 for S'). CONCLUSION: In patients with preserved EF, RVA pacing is associated with the deterioration of both LV diastolic and systolic functions, which is particularly obvious in those with pre-existing LV diastolic dysfunction and V-pace-induced systolic dyssynchrony.

Rho-kinase inhibition: a novel therapeutic target for the treatment of cardiovascular diseases.

The Rho/rho-kinase (ROCK) pathway has an important role in the pathogenesis of several cardiovascular diseases. The activation of ROCK is involved in the regulation of vascular tone, endothelial dysfunction, inflammation and remodeling. The inhibition of ROCK has a beneficial effect in a variety of cardiovascular disorders. Evidence from animal models and from clinical use of ROCK inhibitors, such as Y-27632, fasudil and statins (i.e. pleiotropic effects), supports the hypothesis that ROCK is a potential therapeutic target. This review provides a current understanding of the role of ROCK pathway in the regulation of vascular function and the use of ROCK inhibitors in the treatment of cardiovascular disorders.

A novel method for assessing arterial stiffness by a hydrostatic approach.

Arterial stiffness is an important index for cardiovascular events. The objective of this study is to examine possible parameters related to arterial stiffness that can be estimated during simple arm movements. An experiment was conducted on 32 subjects divided into two groups, one with an age of 26+/-4 years old, and the other 61+/-9. The pulse transit time measured from electrocardiogram to finger photoplethysmogram (PPG) and the amplitude of PPG were calculated beat-to-beat for the subjects while they had their arms lowered. The results of the study showed that the ratio between percentage changes in PTT and finger height are significantly different for the two groups of subjects with different age and health conditions, indicating that parameters can be potentially extracted from this procedure to represent the difference in arterial stiffness of the two groups of subjects.

Incremental prognostic value of combining left ventricular lead position and systolic dyssynchrony in predicting long-term survival after cardiac resynchronization therapy.

The efficacy of CRT (cardiac resynchronization therapy) can be affected by a number of factors; however, the prognostic significance of the LV (left ventricular) lead position has not been explored. The aim of the present study was to examine whether a PL (posterolateral) lead position has an additional value to systolic dyssynchrony in predicting a better survival after CRT. Patients (n=134) who received CRT were followed-up for 39+/-24 months. The LV lead position was determined by cine fluoroscopy, and baseline dyssynchrony was assessed by TDI (tissue Doppler imaging). The relationship between the LV lead position/dyssynchrony and mortality was compared using Kaplan-Meier curves, followed by Cox regression analysis. The all-cause and cardiovascular mortalities were 38 and 31% respectively. The presence of dyssynchrony and a PL lead position predicted a lower all-cause mortality (29 compared with 47%; log-rank chi2=5.38, P=0.02) and cardiovascular mortality (21 compared with 41%; log-rank chi2=6.75, P=0.009) than when absent. The all-cause mortality was as high as 62% when patients had neither dyssynchrony nor a PL lead position, but was reduced to 29% when both criteria were present, and was between 45 and 46% when only one criterion was present (chi2=6.79, P=0.01). The corresponding values for cardiovascular mortality were 62% when patients had neither dyssynchrony nor a PL lead position, 36-38% when patients had either dyssynchrony or a PL lead position, and 21% when patients had both criteria present (chi2=9.54, P=0.004). Combining dyssynchrony and a PL lead position independently predicted a lower all-cause morality {HR (hazard ratio), 0.496 [95% CI (confidence interval), 0.278-0.888]; P=0.018} and cardiovascular mortality [HR, 0.442 (95% CI, 0.232-0.844); P=0.013]. In conclusion, the placement of the LV lead at a PL position provides additional value to baseline dyssynchrony in predicting a lower all-cause and cardiovascular mortality during long-term follow-up after CRT.

Impact of cardiac contractility modulation on left ventricular global and regional function and remodeling.

OBJECTIVES: This study aimed to evaluate the impact of cardiac contractility modulation (CCM) on left ventricular (LV) size and myocardial function. BACKGROUND: CCM is a device-based therapy for patients with advanced heart failure. Previous studies showed that CCM improved symptoms and exercise capacity; however, comprehensive assessment of LV structure, function, and reverse remodeling is not available. METHODS: Thirty patients (60 + or - 11 years, 80% male) with New York Heart Association (NYHA) functional class III heart failure, ejection fraction <35%, and QRS <120 ms were assessed at baseline and 3 months. LV reverse remodeling was measured by real-time 3-dimensional echocardiography. Using tissue Doppler imaging, the peak systolic velocity (Sm) and peak early diastolic velocity (Em) were calculated for LV function, while the standard deviation of the time to peak systolic velocity (Ts-SD) and the time to peak early diastolic velocity (Te-SD) were calculated for mechanical dyssynchrony. RESULTS: LV reverse remodeling was evident, with a reduction in LV end-systolic volume by -11.5 + or - 10.5% and a gain in ejection fraction by 4.8 + or - 3.6% (both p < 0.001). Myocardial contraction was improved in all LV walls, including sites remote from CCM delivery (all p < 0.05); hence, the mean Sm of 12 (2.2 + or - 0.6 cm/s vs. 2.5 + or - 0.7 cm/s) or 6 basal LV segments (2.5 + or - 0.6 cm/s vs. 3.0 + or - 0.7 cm/s) were increased significantly (both p < 0.001). In contrast, CCM had no impact on regional or global Em (2.9 + or - 1.3 cm/s vs. 2.9 + or - 1.1 cm/s), whereas Ts-SD (28.2 + or - 11.2 ms vs. 27.9 + or - 12.7 ms) and Te-SD (30.0 + or - 18.3 ms vs. 30.1 + or - 20.7 ms) remained unchanged (all p = NS). Mitral regurgitation was reduced (22 + or - 14% vs. 17 + or - 15%, p = 0.02). Clinically, there was improvement of NYHA functional class (p < 0.001) and 6-min hall walk distance (p = 0.015). A 24-h Holter monitor showed that premature ventricular contractions were not increased during CCM. CONCLUSIONS: CCM improves both global and regional LV contractility, including regions remote from the impulse delivery, and may contribute to LV reverse remodeling and gain in systolic function. Such improvement is unrelated to diastolic function or mechanical dyssynchrony.

Effect of left ventricular lead concordance to the delayed contraction segment on echocardiographic and clinical outcomes after cardiac resynchronization therapy.

INTRODUCTION: The optimal left ventricular (LV) pacing site for cardiac resynchronization therapy (CRT) is unclear. The current study aims to explore the clinical significance of LV lead concordance to delayed contraction segment in CRT. METHODS AND RESULTS: Concordant LV lead position was defined as the lead tip located by fluoroscopy at or immediately adjacent to the LV segment with latest contraction determined by tissue Doppler imaging. Echocardiographic and clinical outcomes among 101 consecutive patients with or without concordant LV lead positions were compared. There was no significant difference in changes in LV volumes and clinical parameters between patients with concordant (n = 46) or nonconcordant (n = 55) LV lead positions at 3 and 6 months. In multivariate analysis, the baseline asynchrony index (beta= 1.092, 95% CI: 1.050-1.114; P < 0.001), but not LV lead concordance, was the only independent predictor of LV reverse remodeling. By Cox regression analysis, ischemic etiology, and LV reverse remodeling, but not LV lead concordance, were independent predictors of mortality (beta= 2.475, 95% CI: 1.183-5.178; P = 0.016, and beta= 0.272, 95% CI: 0.130-0.567; P < 0.001, respectively), cardiovascular hospitalization (beta= 1.551, 95% CI: 1.032-2.333; P = 0.035, and beta= 0.460, 95% CI: 0.298-0.708; P < 0.001, respectively), and heart failure hospitalization (beta= 0.486, 95% CI: 0.320-0.738; P = 0.001 for LV reverse remodeling). CONCLUSION: LV lead concordance to the delayed contraction segment may not be a major determining factor for favorable echocardiographic and clinical outcomes after CRT.

Reversible left ventricular dyssynchrony and heart failure induced by right ventricular pacing.

Right ventricular (RV) pacing related heart failure is reported in some patients after long term pacing. The exact mechanism is not yet clear but may be related to left ventricular (LV) dyssynchrony induced by RV apical pacing. We report one case with baseline normal LV ejection fraction but complicated by heart failure and ventricular tachycardia after 4 months of pacing for complete heart block together with illustration of LV dyssynchrony demonstrated by tissue Doppler imaging.

Improvement of left atrial function is associated with lower incidence of atrial fibrillation and mortality after cardiac resynchronization therapy.

BACKGROUND: Left atrial (LA) volume is a predictor of cardiovascular events in patients with heart failure. Improvement of LA function and reverse remodeling was observed after cardiac resynchronization therapy (CRT). OBJECTIVE: The purpose of this study was to explore the clinical significance of improvement in LA function after CRT. METHODS: Echocardiographic studies were performed before and 3 months after CRT in 97 patients (72 men and 25 women; age 63.8 +/- 13.3 years) with standard CRT indication but no history of atrial fibrillation (AF). LA active emptying fraction based on the change in volumes (LAV-EF) were calculated, and significant improvement in LA function (LA responder) was defined as a relative increase >/=50% from baseline LAV-EF. The primary end-points were newly developed AF detected by ECG or device and all-cause mortality. RESULTS: After 1,200 +/- 705 days of follow-up, LA responders (n = 47 [48.5%]) had a significantly lower incidence of AF (12.8% vs 40%, P = .002) and mortality (17% vs 44%, P = .004) than did LA nonresponders. In Cox proportional hazard analysis, LA responders was the only independent predictor of lower risk of new-onset AF (hazard ratio 0.22, 95% confidence interval 0.08-0.61, P = .003), whereas both LA responders (hazard ratio 0.22, 95% confidence interval 0.09-0.53, P <.001) and left ventricular reverse remodeling (>10% reduction in left ventricular end-systolic volume at 3 months; hazard ratio 0.96, 95% confidence interval 0.93-0.99, P = .03) were independent predictors of lower risk of death after CRT. CONCLUSION: Improvement of LA function after CRT was associated with a lower incidence of AF and mortality in AF naïve patients with severe heart failure.

Impact of atrial fibrillation in heart failure with normal ejection fraction: a clinical and echocardiographic study.

BACKGROUND: The clinical significance of atrial fibrillation (AF) in heart failure with normal ejection fraction (HFNEF) remains undetermined. METHODS AND RESULTS: We compared the clinical and echocardiographic characteristics among 238 patients hospitalized for HF. Using the cutoff of left ventricular EF of 50%, there were 146 patients with HFNEF (AF = 42) and 92 with systolic HF (AF = 30). When compared among HFNEF, the New York Heart Association (NYHA) class (2.61 +/- 0.51 versus 2.21 +/- 0.46; P < .05), 6-minute walk distance (279.7 +/- 66.0 versus 338.0 +/- 86.1 m; P < .01), quality of life score (26.1 +/- 14.3 versus 19.5 +/- 10.3; P < .05), and previous HF hospitalization were significantly worse in the AF group. These variables were significantly better in HFNEF than systolic HF with sinus rhythm, but the differences were not detected among those with AF. Patients with HFNEF and AF were associated with more severe diastolic dysfunction when compared to sinus rhythm. With a median follow-up of 10.5 months, the proportion of HFNEF patients in AF with recurrent HF hospitalization or death was significantly higher than those in sinus rhythm (28.6% versus 10.6%; P < .01). Both AF and restrictive diastolic dysfunction were independent predictors of HF hospitalization or death in HFNEF. CONCLUSION: Patients with HFNEF and AF were associated with more severe diastolic dysfunction and worse clinical outcomes than those in sinus rhythm.

Improvement of atrial function and atrial reverse remodeling after cardiac resynchronization therapy for heart failure.

OBJECTIVES: We sought to examine whether cardiac resynchronization therapy (CRT) improves atrial function and induces atrial reverse remodeling. BACKGROUND: Cardiac resynchronization therapy is an established therapy for advanced heart failure with prolonged QRS duration, which improves left ventricle (LV) function and is associated with LV reverse remodeling. METHODS: A total of 107 heart failure patients (66 +/- 11 years) who received CRT and were followed up for 3 months were studied. Atrial function was assessed by M-mode, 2-dimensional echocardiography, transmitral Doppler, tissue Doppler velocity, and strain (epsilon) imaging. Left atrial (LA) emptying fraction based on the change in areas (LAA-EF) and volumes (LAV-EF) were calculated. The LV reverse remodeling was defined by a reduction of LV end-systolic volume >10%. RESULTS: In the responders of LV reverse remodeling (n = 62), LAA-EF and LAV-EF were significantly increased (p < 0.001). Responders also had significant decrease in LA size area and volumetric measurements, both before (p < 0.05) and after atrial systole (p < 0.001). However, these parameters were unchanged in the nonresponders (n = 45, p = NS). In the responders, tissue Doppler velocity analysis showed improvement of contraction velocity in both left (p = 0.005) and right atria (p = 0.018), whereas epsilon in both atria were increased in all the phases of cardiac cycle, namely ventricular end-systole (p < 0.001), early diastole (p < 0.001), and late diastole (p = 0.007). CONCLUSIONS: Cardiac resynchronization therapy improves both left and right atrial pump function. The increase in atrial epsilon throughout the cardiac cycle is likely reflecting the improvement of atrial compliance. These changes lead to LA reverse remodeling with reduction of LA size before and after atrial systole.

Effect of left ventricular endocardial activation pattern on echocardiographic and clinical response to cardiac resynchronization therapy.

OBJECTIVE: To explore the left ventricular (LV) electrical activation pattern in heart failure (HF) and its implication to cardiac resynchronization therapy (CRT). DESIGN AND SETTING: Observational study at the University Teaching Hospital. PATIENTS: 23 optimally treated patients with HF with New York Heart Association class III, QRS duration >120 ms and LV ejection fraction <35%. INTERVENTIONS: The LV endocardial activation pattern and total activation time (Tat) was determined by non-contact mapping and the LV mechanical dys-synchrony was determined by standard deviation (Ts-SD) and maximal difference (Ts-diff) of time to peak systolic contraction (Ts) among 12 LV segments using tissue Doppler imaging before receiving CRT. MAIN OUTCOME MEASURES: Correlation between electrical and mechanical dys-synchrony; volumetric responder to CRT at 3 months; HF hospitalisation or death by Kaplan-Meier analysis. RESULTS: Homogenous (type I, n = 8) and presence of conduction block (type II, n = 15) patterns were identified. Significant correlation between Tat and Ts-SD/Ts-diff was noted only in type II (r = 0.73/0.56, p = 0.002/0.03). Ts-SD and Ts-diff in type II were significantly longer than type I. 12 patients in type II and 2 in type I were CRT responders (p = 0.01). After 487 (447) days, patients with type II pattern had significantly lower risk of HF hospitalisation or death than those with type I (log rank chi(2) = 5.25; p = 0.02). CONCLUSION: Patients with type II LV endocardial activation pattern had a more favourable echocardiographic and clinical response to CRT than those with type I pattern.

Diastolic and systolic asynchrony in patients with diastolic heart failure: a common but ignored condition.

OBJECTIVES: The present study aimed to examine whether diastolic and systolic asynchrony exist in diastolic heart failure (DHF) and their prevalence and relationship to systolic heart failure (SHF) patients. BACKGROUND: Few data exist on mechanical asynchrony in DHF. METHODS: Tissue Doppler echocardiography was performed in 373 heart failure patients (281 with SHF and 92 with DHF) and 100 normal subjects. Diastolic and systolic asynchrony was determined by measuring the standard deviation of time to peak myocardial systolic (Ts-SD) and peak early diastolic (Te-SD) velocity using a 6-basal, 6-mid-segmental model, respectively. RESULTS: Both heart failure groups had prolonged Te-SD (DHF vs. SHF vs. controls subjects: 32.2 +/- 18.0 ms vs. 38.0 +/- 25.2 ms vs. 19.5 +/- 7.1 ms) and Ts-SD (31.8 +/- 17.0 ms vs. 36.7 +/- 15.2 ms vs. 17.6 +/- 7.9 ms) compared with the control group (all p < 0.001 vs. control subjects). Based on normal values, the DHF group had comparable diastolic (35.9% vs. 43.1%; chi-square = 1.48, p = NS), but less systolic asynchrony than the SHF group (39.1% vs. 56.9%; chi-square = 8.82, p = 0.003). Normal synchrony, isolated systolic, isolated diastolic, and combined asynchrony were observed in 39.1%, 25.0%, 21.7%, and 14.1% of DHF patients, respectively, and these were 25.6%, 31.3%, 17.4%, and 25.6%, correspondingly, in SHF (chi-square = 10.01, p = 0.019). The correlation between systolic and diastolic asynchrony, and between the myocardial velocities and corresponding mechanical asynchrony appeared weak. A wide QRS duration (>120 ms) was rare in DHF (10.9% vs. 37.7% in SHF) (chi-square = 16.69, p < 0.001). CONCLUSIONS: Diastolic and/or systolic asynchrony was common in 61% of DHF patients despite narrow QRS complex. The presence of asynchrony was not related to myocardial systolic or diastolic function. Systolic and diastolic asynchrony were not tightly coupled, implying distinct mechanisms.