Impact of intravascular ultrasound parameters and platelet reactivity on primary patency after drug-coated balloon angioplasty for femoropopliteal artery disease.

Although the superiority of DCBs to uncoated balloon angioplasty for the treatment of femoropopliteal (FP) lesions has been demonstrated, the association of clinical factors, including anatomical features evaluated by intravascular ultrasound (IVUS) and platelet reactivity, with the loss of patency has not been systematically studied. The current prospective, observational study enrolled 160 consecutive patients (male 67.5%, mean age 74.7 ± 9.7 years) with 213 FP lesions treated with DCBs under IVUS evaluation. The platelet reactivity was measured in P2Y12 reaction units for all of the patients at the DCB treatment. The primary end point was primary patency at 12 months, while the secondary end points were freedom from target lesion revascularization (TLR), all-cause death, major target limb amputation and bleeding events at 12 months. Mean lesion length was 11.9 ± 9.4 cm and 34 (16.0%) were chronic total occlusions (CTOs). Thirty-four (16.0%) were severely calcified lesions. Primary patency by Kaplan-Meier estimate was 79.2% at 12 months, while the 12-month freedom from TLR, all-cause death and bleeding events were observed in 89.1%, 93.4% and 97.4%, respectively. There were no major target limb amputations through 12 months. Multivariate analysis showed that subintimal angioplasty for CTO lesions was a sole risk factor for loss of 12-month primary patency, while other IVUS parameters and platelet reactivity were not.

Multiplexed measurement of cell type-specific calcium kinetics using high-content image analysis combined with targeted gene disruption.

Kinetic analysis of intracellular calcium (Ca2+) in cardiomyocytes is commonly used to determine the pathogenicity of genetic mutations identified in patients with dilated cardiomyopathy (DCM). Conventional methods for measuring Ca2+ kinetics target whole-well cultured cardiomyocytes and therefore lack information concerning individual cells. Results are also affected by heterogeneity in cell populations. Here, we developed an analytical method using CRISPR/Cas9 genome editing combined with high-content image analysis (HCIA) that links cell-by-cell Ca2+ kinetics and immunofluorescence images in thousands of cardiomyocytes at a time. After transfecting cultured mouse cardiomyocytes that constitutively express Cas9 with gRNAs, we detected a prolonged action potential duration specifically in Serca2a-depleted ventricular cardiomyocytes in mixed culture. To determine the phenotypic effect of a frameshift mutation in PKD1 in a patient with DCM, we introduced the mutation into Cas9-expressing cardiomyocytes by gRNA transfection and found that it decreases the expression of PKD1-encoded PC1 protein that co-localizes specifically with Serca2a and L-type voltage-gated calcium channels. We also detected the suppression of Ca2+ amplitude in ventricular cardiomyocytes with decreased PC1 expression in mixed culture. Our HCIA method provides comprehensive kinetic and static information on individual cardiomyocytes and allows the pathogenicity of mutations to be determined rapidly.

Infrapopliteal Anatomic Severity and Delayed Wound Healing in Patients With Chronic Limb-Threatening Ischemia in the Era of the Global Limb Anatomic Staging System.

Purpose: To investigate the prognostic impact of infrapopliteal (IP) artery anatomic severity according to the Global Limb Anatomic Staging System (GLASS) on delayed wound healing in patients with chronic limb-threatening ischemia (CLTI). Materials and Methods: This study retrospectively analyzed 639 limbs with tissue loss in 484 CLTI patients (mean age 74±10 years; 300 men) presenting IP lesions treated with endovascular therapy between April 2010 and December 2015. Two-thirds of patients had diabetes (323, 67%) and over half were on hemodialysis (255, 53%). More than a third of the limbs (251, 39%) were classified as clinical stage 4 according to the Wound, Ischemia, and foot Infection (WIfI) system. IP anatomic severity was classified based on preprocedural angiography according to the GLASS. Severity of arterial calcification was assessed using high-intensity fluoroscopy and classified into 3 groups: none (grade 0), unilateral (grade 1), and bilateral (grade 2). Poor below-the-ankle (BTA) runoff was defined as the lack of a pedal arch with 0 to 1-vessel runoff within the dorsal pedis artery and the lateral and medial plantar arteries. The outcome measure was 1-year wound healing. The association of anatomic characteristics with delayed wound healing was evaluated using Cox proportional hazards regression analysis. Outcomes are presented as the adjusted hazard ratio (HR) with 95% confidence interval (CI). Results: During a mean follow-up of 22±19 months, the 1-year cumulative wound healing rate was estimated to be 59.0% (95% CI 54.5% to 63.5%). Multivariable analysis demonstrated independent associations between delayed wound healing and IP calcification grade (HR 1.24, 95% CI 1.02 to 1.50, p=0.027) and poor BTA runoff (HR 1.39, 95% CI 1.04 to 1.85, p=0.025) but not the GLASS IP grade (HR 0.92, 95% CI 0.82 to 1.14, p=0.21). Conclusion: The current study revealed that IP arterial calcification and poor BTA runoff were significantly associated with delayed wound healing, whereas the GLASS was not predictive of wound healing.

Impact of the Temporal Relationship Between Atrial Fibrillation and Heart Failure on Prognosis After Ablation.

BACKGROUND: The purpose of this study was to elucidate the effect of the temporal relationship between atrial fibrillation (AF) and heart failure (HF) on clinical outcomes after catheter ablation.Methods and Results:We included 129 consecutive patients with AF and HF who underwent catheter ablation in hospital from December 2014 to September 2017. The patients were divided into 2 groups based on the temporal relationship between AF and HF. Group 1 consisted of 42 patients with AF following HF while Group 2 consisted of 87 patients with AF preceding HF or those who developed both of them simultaneously at the timing of first visit to a doctor. The primary endpoint was a composite of death and hospitalization due to HF during a 2-year follow-up. AF recurrence was more common in Group 1 (45% vs. 23%; hazard ratio [HR], 2.49; 95% confidence interval [CI], 1.25-4.94; P=0.009). Death and HF hospitalization were more frequent in Group 1 (19 [45%], 6 [7%] patients, respectively, P<0.0001). After adjustment for several covariates, patients in Group 1 were independently associated with poorer outcomes after AF ablation (HR, 8.66; 95% CI, 2.942-5.5; P<0.0001). CONCLUSIONS: Adverse clinical outcomes of death, HF hospitalization and AF recurrence were more frequent in patients with AF following HF than in those with AF preceding HF.

A saline contrast-enhanced echocardiography-guided approach to cryoballoon ablation.

BACKGROUND: Pulmonary vein isolation (PVI) with a cryoballoon usually uses iodinated contrast medium for confirmation of venous occlusion. We hypothesized that an alternative to angiography with iodinated contrast, saline bolus-enhanced echocardiography with the microbubbles formed in situ, could be used to assess venous occlusion. We sought to assess the efficacy and safety of the contrast-enhanced intracardiac echocardiographic (ICE) approach. METHODS: Thirty-four consecutive patients without iodine sensitivity or renal insufficiency were studied to assess the accuracy of ICE-guided PV occlusion underwent both angiographic guidance and ICE guidance (validation group). Twenty consecutive patients with paroxysmal atrial fibrillation (AF) and contraindications to iodinated contrast medium (ICE-guided group) underwent PV occlusion and ablation with ICE guidance alone. Procedural results and clinical outcomes were compared with those of 245 control patients undergoing PVI by the conventional angiographic method (control group). RESULTS: In the validation group, ICE-guided PV occlusion was as effective as angiography-guided PV occlusion. In the ICE-guided group, two patients required touch-up ablation using a radiofrequency ablation catheter with fluoroscopic guidance. Procedure time, radiation exposure, and requirements for touch-up ablation were similar between the ICE-guided group and the control group. In patients requiring only PVI, the ICE-guided approach involved significantly less radiation exposure than the conventional approach. There was no significant difference in atrial fibrillation (AF)-free survival rate between the two groups during a follow-up period of 14 ± 6 months. CONCLUSIONS: An echo-guided approach using saline infusion was effective in terms of avoidance of iodinated contrast use and radiation exposure.

Efficacy and safety of the cryoballoon-based atrial fibrillation ablation in patients aged ≥80 years.

INTRODUCTION: The prevalence of atrial fibrillation (AF) increases with age. Although the efficacy of cryoballoon ablation (CB-A) in patients with AF has been demonstrated, data on the efficacy of this CB-A in elderly patients are limited. The aim of this study was to evaluate the 1-year efficacy and safety of CB-A in patients aged ≥80 years compared with those less than 80 years. METHODS: A total of 49 consecutive patients ≥80 years with paroxysmal AF (PAF) who underwent CB-A were compared with 241 patients aged <80 years. AF-free survival rate at 1 year and procedural data were compared between the groups. RESULTS: Median (25th, 75th quartile) age was 83 (82, 85) years in the older group and 68 (61, 73) years in the younger group. At 1-year follow-up, the success rate did not significantly differ between the groups. In Cox regression analysis, age ≥80 years was not significantly related to AF recurrence. Low-voltage areas were more frequently observed in the elderly group than the control group (39% vs 17%; P = .01). Fluoroscopy time and procedure time did not differ significantly. There was no significant difference in the occurrence rate of transient phrenic nerve palsy. No severe complications occurred in either group, including procedure-related deaths, atrioesophageal fistula, cardiac tamponade, and cerebrovascular embolic events. CONCLUSION: The results of our study showed that CB-A for PAF is a feasible and safe procedure even in elderly patients, with similar success and complication rates when compared with a younger population.

Left atrial voltage mapping with a direction-independent grid catheter: Comparison with a conventional circular mapping catheter.

INTRODUCTION: A recently introduced grid mapping catheter (GMC) is designed for better electrode-tissue contact and can collect bipolar signals both along and across the splines, which may allow more efficient voltage map generation independent of propagation direction. We compared the GMC with a conventional circular mapping catheter (CMC) for left atrial (LA) voltage mapping. METHODS: This study included 20 consecutive patients undergoing repeat ablation for recurrent atrial fibrillation who had demonstrated LA low-voltage areas (LVAs, <0.10 mV). Following pulmonary vein isolation, LA voltage mapping was performed twice, once using the GMC and once using the CMC. RESULTS: Voltage mapping was more efficient using the GMC than the CMC in terms of mapping time (459 [404, 543] vs 602 [496, 814] seconds; P = .014) and the number of mapping points (2446 [2099, 3104] vs 1841 [1494, 2314]; P = .002). The incidence of catheter-induced ectopies was lower (44 [28, 62] vs 114 [74, 188]; P < .0001) using the GMC. The GMC utilizing all bipoles detected LVAs in 85% of patients with LVAs detected by CMC. LVA measurements were significantly smaller on maps generated by the GMC using bipoles along or across the splines than those measured with the CMC (11.1 [4.6, 17.2] or 9.7 [2.5, 16.0] vs 16.4 [6.8, 26.8] cm2 ; P = .008 and P = .001, respectively), and were even smaller when using all bipoles (7.9 [1.1, 13.5] cm2 , P = .0001). CONCLUSION: The GMC allowed a more efficient mapping procedure and enabled more selective identification of LVAs with smaller LVA size.

Impact of Renal Dysfunction on Left Atrial Low-Voltage Areas in Patients With Atrial Fibrillation.

BACKGROUND: The presence of residual left atrial low-voltage areas (LVA) has been shown to be strongly associated with atrial fibrillation (AF) recurrence after pulmonary vein isolation. A preliminary study showed that concomitant chronic kidney disease (CKD) increased the rate of AF recurrence. The association between CKD and LVA, however, has not been elucidated. In the present study, we investigated the association between CKD severity and LVA prevalence. Methods and Results: In total, 183 consecutive AF patients who underwent initial ablation for AF were enrolled in this retrospective observational study. Serum cystatin C before ablation was measured, and the estimated glomerular filtration rate (eGFR) was calculated. LVA were defined as sites of left atrial electrogram amplitude <0.5 mV. Of 183 patients, 76 (42%) had LVA. Patients with LVA had lower eGFR calculated using cystatin C (74±22 vs. 86±24 mL/min/1.73 m2, P=0.001). The optimal cut-off of the calculated eGFR was 71.5 mL/min/1.73 m2, corresponding to a 79.4% sensitivity, 50% specificity, and 67.2% predictive accuracy. LVA occurred more frequently in patients with more severe categories of CKD. On multivariate analysis, eGFR <71.5 mL/min/1.73 m2was an independent predictor of LVA (odds ratio, 3.3; 95% CI: 1.4-7.8; P=0.006). CONCLUSIONS: CKD severity was correlated with left atrial LVA prevalence in patients with AF undergoing catheter ablation.

Comparison of electrogram waveforms between a multielectrode mapping catheter and a linear ablation catheter.

BACKGROUND: Smaller low-voltage areas (LVAs) obtained by multielectrode catheters were reported than those by linear ablation catheters. However, the underlying electrogram difference has not been elucidated. This study aimed to compare the two mapping catheters' measurements of electrogram waveforms and LVAs. METHODS: This prospective observational study included 17 consecutive patients undergoing ablation for persistent atrial fibrillation. Following pulmonary vein isolation, voltage mapping during sinus rhythm was performed once using the ablation catheter, and once using the multielectrode catheter. Approximately 20 pairs of mapping points at approximately the same position between the two voltage maps were manually selected evenly throughout the left atrium. RESULTS: Voltage mapping with the multielectrode catheter demonstrated smaller LVAs, defined as <0.50 mV (5.9 [3.3, 11.0] vs 9.7 [6.6, 16.9] cm2 ) than those mapped with the ablation catheter. The two mapping catheters' voltage amplitudes of all pairs of mapping points correlated well (r = 0.81, P < 0.0001) overall, but they did not correlate within diseased areas (either voltage <0.50 mV). The voltage amplitude difference between the two catheters ([VolMulti  - VolAbl ]/VolAbl × 100) was greater in the diseased areas (37.4% [-9.8%, 147%]) than in the healthy areas (both voltages ≥0.50 mV, 26.2% [-13.0%, 92.8%], P = 0.014). The electrogram waveform of the multiple electrode catheter displayed a higher voltage amplitude, shorter duration, greater number of peaks, and lower dull peak ratio (number of dull peaks/total peaks) than that of the ablation catheter. CONCLUSION: The multielectrode catheter produced smaller LVA measurements with sharper and higher voltage electrograms compared to the ablation catheter, specifically in diseased areas.

Comparison of Clinical Outcomes between Endovascular Therapy with Self-Expandable Nitinol Stent and Femoral-Popliteal Bypass for Trans-Atlantic Inter-Society Consensus II C and D Femoropopliteal Lesions.

BACKGROUND: Femoral-popliteal (F-P) bypass is the standard treatment for lower extremity peripheral artery disease with severe femoropopliteal (FP) artery lesions. However, in clinical settings, these patients are treated with endovascular therapy (EVT) because of frailty and difficulties with general anesthesia. We compared the clinical outcomes of F-P bypass and EVT for severe FP artery lesions and investigated the types of patients for whom EVT would be as appropriate as F-P bypass. METHODS: This multicenter, retrospective study included 452 Trans-Atlantic Inter-Society Consensus (TASC) II C and D FP artery lesions in 352 patients. A total of 350 lesions in 260 patients (74 ± 9 years, 66% male, 60% claudication) were treated with EVT with nitinol self-expandable stents, and 102 lesions in 92 patients (71 ± 9 years, 75% male, 40% claudication) were treated with F-P bypass. The primary outcome measure was primary patency, and the influence of baseline characteristics on its association with the treatment strategy (EVT versus F-P bypass) was assessed using a Cox proportional hazards regression model. RESULTS: Kaplan-Meier analysis indicated that the 3-year primary patency rate was significantly lower for EVT than F-P bypass (60% vs. 74%, P < 0.01). The body mass index (BMI) and C-reactive protein (CRP) levels significantly interacted with the treatment strategy for restenosis (P < 0.05). The adjusted hazard ratios of EVT versus F-P bypass for restenosis were 0.77 (P = 0.46) in cases with a low BMI (≤18 kg/m2) or an elevated CRP level (≥1 mg/dL) and 3.35 (P < 0.01) in other cases. The 3-year primary patency rate was not significantly different between the EVT and F-P bypass groups in patients with BMI ≤ 18 kg/m2 or CRP ≥ 1 mg/dL (57% vs. 45%, P = 0.84). CONCLUSIONS: In TASC II C and D lesions, EVT appears to yield patency comparable to that of F-P bypass in patients with a low BMI or an elevated CRP level, but lower patency in other patients.

Association of Subclinical Intrastent Thrombus Detected 9 Months After Implantation of 2nd-Generation Drug-Eluting Stent With Future Major Adverse Cardiac Events　- A Coronary Angioscopic Study.

BACKGROUND: Detection of yellow plaques (YP) by coronary angioscopy (CAS) 1 year after 1st-generation drug-eluting stent (DES) implantation has been related to future coronary events. However, the association between CAS findings and clinical outcomes following 2nd-generation DES implantation has not been investigated. Methods and Results: This study included a total of 248 2nd-generation DES in 179 patients, who were examined by CAS 9±2 months after implantation. Angioscopic evaluation included dominant neointimal coverage (NIC) grade, heterogeneity of NIC, presences of YP and intrastent thrombus. The outcome measure was major adverse cardiac events (MACE) defined as a composite of cardiac death, acute myocardial infarction and any coronary revascularization. The association between the CAS findings and MACE was evaluated using the Kaplan-Meier method. A Cox proportional hazards model was used to assess the predictors of MACE. The mean follow-up duration was 1,367±843 days. Dominant NIC grade (P=0.98), heterogeneity of NIC (P=0.20) and YP (P=0.53) were not associated with the incidence of MACE. However, intrastent thrombus was significantly associated with MACE (P=0.033). Intrastent thrombus (adjusted hazard ratio: 2.22; 95% confidence interval [CI]: 1.12-4.39), acute coronary syndrome (2.83; 95% CI: 1.42-5.67) and B2/C lesion (2.13; CI: 1.12-4.05) were independent predictors of MACE. CONCLUSIONS: Subclinical intrastent thrombus observed by CAS at 9 months after 2nd-generation DES implantation was independently associated with poor clinical outcome.

A novel predictor of clinical outcomes in patients with heart failure with preserved left-ventricular ejection fraction: a pilot study.

Predictors of outcomes in patients with heart failure with preserved left-ventricular ejection fraction (HFpEF) remain unclear. The ratio of early diastolic transmitral flow velocity to early diastolic myocardial velocity (E/e') has been proposed, but the predictive accuracy remains unsatisfactory. We hypothesized that E/e' normalized by the stroke volume (SV), E/e'/SV, could be a good predictor of outcome in HFpEF patients by reflecting the terminal slope of the end-diastolic pressure-volume relation, i.e., stiffness of the left ventricle. This pilot study was conducted to propose a novel echocardiographic index for predicting the outcomes of patients with HFpEF. Echocardiography including E/e'/SV measurement was performed in consecutive 80 HFpEF patients at discharge in index hospitalization due to HF. The end points included the readmission for heart failure and cardiovascular death at 1 year after discharge. 19 patients (24%) met the end points. Receiver-operating characteristic analysis showed that E/e''/SV was a strong predictive factor (AUC = 0.78) compared to E/e' (AUC = 0.74). Kaplan-Meier analysis showed that patients with E/e'/SV > 0.40 had a poorer prognosis than those with E/e'/SV < 0.40 (p < 0.01). By Cox regression multi-variate analysis, a high E/e'/SV was an independent predictor of event-free survival [adjusted hazard ratio (95% CI) 14.26 (3.18, 63.93) (p = 0.01)]. E/e'/SV has potential to predict clinical outcomes in patients with HFpEF.

Impact of Calcification on Clinical Outcomes After Endovascular Therapy for Superficial Femoral Artery Disease: Assessment Using the Peripheral Artery Calcification Scoring System.

PURPOSE: To investigate whether the severity of lesion calcification assessed by the novel peripheral artery calcification scoring system (PACSS) was associated with clinical outcomes after endovascular therapy (EVT) for superficial femoral artery (SFA) lesions. METHODS: A retrospective analysis was conducted of 394 consecutive patients (mean age 72±8 years; 290 men) with intermittent claudication [223 (57%) with diabetes, 81 (21%) on hemodialysis] who underwent successful EVT for de novo SFA lesions [length 152.1±95.7 mm; 199 (50%) TransAtlantic Inter-Society Consensus II class C/D] between January 2010 and December 2013. The patients were retrospectively categorized using the PACSS classification (grades 0-4: no visible calcification of the target lesion, unilateral wall calcification <5 cm, unilateral calcification ≥5 cm, bilateral wall calcification <5 cm, and bilateral calcification ≥5 cm, respectively). The main outcome was primary patency, while the secondary outcome measures were mortality and major adverse limb events [MALE: any intervention (repeat EVT or surgical revision) or major (above ankle) amputation]. Cox proportional hazards analysis was used to explore whether the PACSS classification was an independent predictor of clinical outcomes. Results are presented as the hazard ratio (HR) and 95% confidence interval (CI). RESULTS: The distribution of PACSS grades was 0 in 54%, grade 1 in 16%, grade 2 in 12%, grade 3 in 9%, and grade 4 in 9%. The 2-year primary patency rates in these grades, respectively, were 70.0%, 66.6%, 72.1%, 55.6%, and 36.3% (p<0.001). After multivariate analysis, PACSS grade 4 (HR 2.74, 95% CI 1.56 to 4.83, p<0.001), diabetes (HR 1.52, 95% CI 1.06 to 2.20, p=0.022), lesion length (HR 1.04, 95% CI 1.01 to 1.07, p=0.006), and vessel diameter (HR 0.80, 85% CI 0.65 to 0.98, p=0.038) were associated with loss of primary patency. PACSS grade 4 was also associated with MALE and mortality (p=0.048 and 0.011, respectively). Bare metal stent use (HR 0.47, 95% CI 0.30 to 0.73, p<0.001) was positively associated with primary patency. CONCLUSION: PACSS grade 4 calcification was independently associated with clinical outcomes after EVT for de novo SFA lesions.

Modeling Reduced Contractility and Stiffness Using iPSC-Derived Cardiomyocytes Generated From Female Becker Muscular Dystrophy Carrier.

Study investigators encountered a female Becker muscular dystrophy (BMD) carrier with advanced heart failure (HF) and identified a stop-gain variant in procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 (PLOD3) as a potential second-hit variant. Isogenic induced pluripotent stem cells (iPSCs) with dominant expression of WT-DMD, Δ45-48-DMD, or Δ45-48-DMD with corrected PLOD3 variant were established. Microforce testing using 3-dimensional self-organized tissue rings (SOTRs) generated from iPSC-derived cardiomyocytes (iPSC-CMs) demonstrated that correction of the heterozygous PLOD3 variant did not improve the reduced force, but it significantly recovered the reduced stiffness in Δ45-48-DMD SOTRs. Correction of the PLOD3 variant restored collagen synthesis in iPSC-CMs. Our findings revealed the pathogenesis underlying advanced HF in a female BMD carrier.

SARS-CoV-2 spike receptor-binding domain is internalized and promotes protein ISGylation in human induced pluripotent stem cell-derived cardiomyocytes.

Although an increased risk of myocarditis has been observed after vaccination with mRNA encoding severe acute respiratory syndrome coronavirus 2 spike protein, its underlying mechanism has not been elucidated. This study investigated the direct effects of spike receptor-binding domain (S-RBD) on human cardiomyocytes differentiated from induced pluripotent stem cells (iPSC-CMs). Immunostaining experiments using ACE2 wild-type (WT) and knockout (KO) iPSC-CMs treated with purified S-RBD demonstrated that S-RBD was bound to ACE2 and internalized into the subcellular space in the iPSC-CMs, depending on ACE2. Immunostaining combined with live cell imaging using a recombinant S-RBD fused to the superfolder GFP (S-RBD-sfGFP) demonstrated that S-RBD was bound to the cell membrane, co-localized with RAB5A, and then delivered from the endosomes to the lysosomes in iPSC-CMs. Quantitative PCR array analysis followed by single cell RNA sequence analysis clarified that S-RBD-sfGFP treatment significantly upregulated the NF-kß pathway-related gene (CXCL1) in the differentiated non-cardiomyocytes, while upregulated interferon (IFN)-responsive genes (IFI6, ISG15, and IFITM3) in the matured cardiomyocytes. S-RBD-sfGFP treatment promoted protein ISGylation, an ISG15-mediated post-translational modification in ACE2-WT-iPSC-CMs, which was suppressed in ACE2-KO-iPSC-CMs. Our experimental study demonstrates that S-RBD is internalized through the endolysosomal pathway, which upregulates IFN-responsive genes and promotes ISGylation in the iPSC-CMs.

Impact of Longer Hemodialysis Vintage with Higher Serum Phosphorus Level on Clinical Outcomes in Patients with Chronic Limb-Threatening Ischemia Presenting Tissue Loss after Endovascular Therapy.

AIMS: Hemodialysis vintage and serum phosphorus levels adversely affect outcomes in patients on hemodialysis. Whether these factors have a similar prognostic impact on patients who are on hemodialysis and have chronic limb-threatening ischemia (CLTI) has not been systematically studied. We aimed to explore the risk factors, including hemodialysis vintage and serum phosphorus levels, on clinical outcomes after endovascular therapy (EVT) in hemodialysis patients with CLTI. METHODS: The current study rerospectively analyzed 374 hemodialysis patients with CLTI presenting with ischemic tissue loss (age: 72.3±9.0 years, male: 73.3%, diabetes mellitus: 68.2%, Rutherford 5: 75.9%, 6: 24.1%, WIfI stage 4: 50.0%) primarily treated with EVT between April 2007 and December 2016. The primary outcome measure was 1-year amputation-free survival (AFS), while the secondary outcome measure was 1-year wound healing. Predictors for each outcome were evaluated by Cox proportional hazards model. RESULTS: Multivariate analysis significantly associated longer hemodialysis vintages with higher serum phosphorus levels (hazard ratio [HR], 0.599; 95% confidence interval [CI], 0.394-0.910; p=0.016) with 1-year AFS. Longer vintages for hemodialysis with higher serum phosphorus levels were marginally, but not significantly, associated with 1-year wound healing. (HR, 0.684; 95% CI, 0.467-1.000; p=0.050). CONCLUSION: Longer hemodialysis vintages with higher serum phosphorus levels adversely affect outcomes after EVT for hemodialysis patients with CLTI presenting with ischemic tissue loss.

Modeling reduced contractility and impaired desmosome assembly due to plakophilin-2 deficiency using isogenic iPS cell-derived cardiomyocytes.

Loss-of-function mutations in PKP2, which encodes plakophilin-2, cause arrhythmogenic cardiomyopathy (AC). Restoration of deficient molecules can serve as upstream therapy, thereby requiring a human model that recapitulates disease pathology and provides distinct readouts in phenotypic analysis for proof of concept for gene replacement therapy. Here, we generated isogenic induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) with precisely adjusted expression of plakophilin-2 from a patient with AC carrying a heterozygous frameshift PKP2 mutation. After monolayer differentiation, plakophilin-2 deficiency led to reduced contractility, disrupted intercalated disc structures, and impaired desmosome assembly in iPSC-CMs. Allele-specific fluorescent labeling of endogenous DSG2 encoding desmoglein-2 in the generated isogenic lines enabled real-time desmosome-imaging under an adjusted dose of plakophilin-2. Adeno-associated virus-mediated gene replacement of PKP2 recovered contractility and restored desmosome assembly, which was sequentially captured by desmosome-imaging in plakophilin-2-deficient iPSC-CMs. Our isogenic set of iPSC-CMs recapitulates AC pathology and provides a rapid and convenient cellular platform for therapeutic development.

Human-Induced Pluripotent Stem Cell-Derived Cardiomyocyte Model for TNNT2 Δ160E-Induced Cardiomyopathy.

BACKGROUND: The Δ160E mutation in TNNT2, which encodes troponin T, is a rare pathogenic variant identified in patients with hypertrophic cardiomyopathy and is associated with poor prognosis. Thus, a convenient human model recapitulating the pathological phenotype caused by TNNT2 Δ160E is required for therapeutic development. METHODS: We identified a heterozygous in-frame deletion mutation (c.478_480del, p.Δ160E) in TNNT2 in a patient with familial hypertrophic cardiomyopathy showing progressive left ventricular systolic dysfunction, leading to advanced heart failure. To investigate the pathological phenotype caused by Δ160E, we generated a set of isogenic induced pluripotent stem cells carrying the heterozygous Δ160E, homozygously corrected or homozygously introduced Δ160E using genome editing and differentiated them into cardiomyocytes (Hetero-Δ160E-, wild type-, and Homo-Δ160E-induced pluripotent stem cells [iPSC]-derived cardiomyocytes [iPSC-CMs]). RESULTS: Hetero-Δ160E-iPSC-CMs exhibited prolonged calcium decay, relaxation impairment, and hypertrophy compared to wild type-iPSC-CMs. Notably, these phenotypes were further exacerbated in Homo-Δ160E-iPSC-CMs. Overexpression of R-GECO-fused Δ160E mutant troponin T prolonged decay time and time to peak of the myofilament-localized calcium transient in iPSC-CMs, indicating that sarcomeric calcium retention with Δ160E may affect intracellular calcium concentration. High-content imaging analysis detected remarkable nuclear translocation of NFATc1, especially in Homo-Δ160E-iPSC-CMs, indicating that the Δ160E mutation promotes hypertrophic signaling pathway in a dose-dependent manner. Increased phosphorylation of CaMKIIδ (calcium/calmodulin-dependent protein kinase IIδ) and phospholamban at Thr17 was observed in Homo- and Hetero-Δ160E-iPSC-CMs. Epigallocatechin-3-gallate, a calcium desensitizing compound, shortened prolonged calcium decay and relaxation duration in Δ160E-iPSC-CMs. CONCLUSIONS: Isogenic iPSC-CMs recapitulate the prolonged calcium decay, relaxation impairment, and subsequent calcium-regulated signaling pathways caused by the TNNT2 Δ160E mutation and can serve as a human model for therapeutic development to prevent hypertrophic cardiomyopathy pathology.

Long-Term Outcomes and Clinical Predictors of Mortality Following Occurrence of Stent Thrombosis.

Background Stent thrombosis (ST) remains a significant medical issue. In particular, longer-term mortality and clinical predictors after ST occurrence have yet to be elucidated. Methods and Results This was a multicenter, retrospective, observational study. A total of 187 definite ST cases from January 2008 to December 2017 were enrolled, and the long-term clinical outcomes were investigated. The primary outcome measure was the cumulative mortality after ST occurrence. In addition, independent predictors of mortality were assessed. Among the stent types causing ST, bare-metal stent, first-generation drug-eluting stent, second-generation drug-eluting stent, and third-generation drug-eluting stent comprised 31.0%, 19.3%, 36.9%, and 6.4% of cases, respectively. Median duration from stent implantation to ST was 680.5 (interquartile range, 33.8-2450.5) days. Cumulative mortality was 14.6%, 17.4%, 21.2%, 24.4%, and 33.8% at 1, 2, 3, 5 and 10 years, respectively. The cumulative mortality did not significantly differ by type of stent, and mortality of late ST was higher than that of early ST and very late ST; however, it did not reach statistical significance after the multivariate analysis. Independent predictors of mortality were hemodialysis (hazard ratio [HR], 7.80; 95% CI, 3.07-19.81; P<0.001), culprit lesions in the left main trunk (HR, 8.14; 95% CI, 1.71-38.75; P=0.008), culprit lesions in the left coronary artery (HR, 2.77; 95% CI, 1.10-6.96; P=0.030), and peak creatine kinase (HR, 1.017; 95% CI, 1.011-1.022; P<0.001). Conclusions The 10-year cumulative mortality after ST reached 33.8%. Close follow-up is thus mandatory for patients with ST, especially with hemodialysis, culprit lesions in the left main trunk and left coronary artery, and high peak creatine kinase.

Effect of Rehabilitation Nutrition Care Process on Physical Function in Lung Cancer Cachexia: A Case Report.

OBJECTIVES: Patients with cancer cachexia have poor adherence to treatment, which affects their prognosis. Currently, there are many studies on the effects of rehabilitation on cancer cachexia, but there is a lack of evidence on the effects of nutrition therapy alone or in combination with rehabilitation and nutrition therapy. This article describes a case in which rehabilitation nutrition care process was effective in a patient with lung cancer who developed cancer cachexia. METHODS: A 68-year-old woman was hospitalized for treatment of lung adenocarcinoma. The patient had moderate malnutrition, sarcopenia, and cachexia at the time of admission, so the authors intervened according to rehabilitation nutrition care process. The physiotherapist mainly prescribed resistance training and aerobic exercise, 40-60 minutes a day, 5-6 days a week. And the dietitian provided oral nutritional supplements (100 kcal, branched-chain amino acid: 3.0 g) in addition to hospital food and adjusted the patient's energy intake to 26.96-33.05 kcal/kg/day and protein intake to 1.07-1.14 g/kg/day. OUTCOMES: Comparing the initial evaluation with the discharge, nutritional status, such as body mass index and skeletal muscle mass, and physical functions, such as maximum grip strength, gait speed, and functional independence measure (motor items), were improved. CONCLUSIONS: Rehabilitation nutrition care process-based interventions may improve nutritional status and physical functions more than exercise therapy alone in patients with lung cancer cachexia.