Clinical, Echocardiographic, and Longitudinal Characteristics Associated With Heart Failure With Improved Ejection Fraction.

Heart failure with improved ejection fraction (HFimpEF) has better outcomes than HF with reduced EF (HFrEF). However, factors contributing to HFimpEF remain unclear. This study aimed to evaluate clinical and longitudinal characteristics associated with subsequent HFimpEF. This was a single-center retrospective HFrEF cohort study. Data were collected from 2014 to 2022. Patients with HFrEF were identified using International Classification of Diseases codes, echocardiographic data, and natriuretic peptide levels. The main end points were HFimpEF (defined as EF >40% at ≥3 months with ≥10% increase) and mortality. Cox proportional hazards and mixed effects models were used for analyses. The study included 1,307 patients with HFrEF with a median follow-up of 16.3 months (interquartile range 8.0 to 30.6). The median age was 65 years; 68% were male whereas 57% were White. On follow-up, 38.7% (n = 506) developed HFimpEF, whereas 61.3% (n = 801) had persistent HFrEF. A multivariate Cox regression model identified gender, race, co-morbidities, echocardiographic, and natriuretic peptide as significant covariates of HFimpEF (p <0.05). The HFimpEF group had better survival compared with the persistent HFrEF group (p <0.001). Echocardiographic and laboratory trajectories differed between groups. In this HFrEF cohort, 38.7% transitioned to HFimpEF and approximately 50% met the definition within the first 12 months. In a HFimpEF model, gender, co-morbidities, echocardiographic parameters, and natriuretic peptide were associated with subsequent HFimpEF. The model has the potential to identify patients at risk of subsequent persistent or improved HFrEF, thus informing the design and implementation of targeted quality-of-care improvement interventions.

Clinical, Echocardiographic, and Longitudinal Characteristics Associated with Heart Failure with Improved Ejection Fraction.

Background: Heart failure (HF) with improved ejection fraction (HFimpEF) has better outcomes than HF with reduced ejection fraction (HFrEF). However, factors contributing to HFimpEF remain unclear. This study aimed to evaluate clinical and longitudinal characteristics associated with subsequent HFimpEF. Methods: This was a single-center retrospective HFrEF cohort study. Data were collected from 2014 to 2022. Patients with HFrEF were identified using ICD codes, echocardiographic data, and natriuretic peptide levels. The main endpoints were HFimpEF (defined as ejection fraction >40% at ≥3 months with ≥10% increase) and mortality. Cox proportional hazards and mixed effects models were used for analyses. Results: The study included 1307 HFrEF patients with a median follow-up of 16.3 months (IQR 8.0-30.6). The median age was 65 years; 68% were male while 57% were white. On follow-up, 39% (n=506) developed HFimpEF, while 61% (n=801) had persistent HFrEF. A multivariate Cox regression model identified sex, race comorbidities, echocardiographic, and natriuretic peptide as significant covariates of HFimpEF ( p <0.05). The HFimpEF group had better survival compared to the persistent HFrEF group ( p <0.001). Echocardiographic and laboratory trajectories differed between groups. Conclusion: In this HFrEF cohort, 39% transitioned to HFimpEF and approximately 50% met the definition within the first 12 months. In a HFimpEF model, sex, comorbidities, echocardiographic parameters, and natriuretic peptide were associated with subsequent HFimpEF. The model has the potential to identify patients at risk of subsequent persistent or improved HFrEF, thus informing the design and implementation of targeted quality-of-care improvement interventions.

MapperPlus: Agnostic clustering of high-dimension data for precision medicine.

One of the goals of precision medicine is to classify patients into subgroups that differ in their susceptibility and response to a disease, thereby enabling tailored treatments for each subgroup. Therefore, there is a great need to identify distinctive clusters of patients from patient data. There are three key challenges to three key challenges of patient stratification: 1) the unknown number of clusters, 2) the need for assessing cluster validity, and 3) the clinical interpretability. We developed MapperPlus, a novel unsupervised clustering pipeline, that directly addresses these challenges. It extends the topological Mapper technique and blends it with two random-walk algorithms to automatically detect disjoint subgroups in patient data. We demonstrate that MapperPlus outperforms traditional agnostic clustering methods in key accuracy/performance metrics by testing its performance on publicly available medical and non-medical data set. We also demonstrate the predictive power of MapperPlus in a medical dataset of pediatric stem cell transplant patients where a number of cluster is unknown. Here, MapperPlus stratifies the patient population into clusters with distinctive survival rates. The MapperPlus software is open-source and publicly available.

Total-Body Perfusion Imaging with [11C]-Butanol.

Tissue perfusion can be affected by physiology or disease. With the advent of total-body PET, quantitative measurement of perfusion across the entire body is possible. [11C]-butanol is a perfusion tracer with a superior extraction fraction compared with [15O]-water and [13N]-ammonia. To develop the methodology for total-body perfusion imaging, a pilot study using [11C]-butanol on the uEXPLORER total-body PET/CT scanner was conducted. Methods: Eight participants (6 healthy volunteers and 2 patients with peripheral vascular disease [PVD]) were injected with a bolus of [11C]-butanol and underwent 30-min dynamic acquisitions. Three healthy volunteers underwent repeat studies at rest (baseline) to assess test-retest reproducibility; 1 volunteer underwent paired rest and cold pressor test (CPT) studies. Changes in perfusion were measured in the paired rest-CPT study. For PVD patients, local changes in perfusion were investigated and correlated with patient medical history. Regional and parametric kinetic analysis methods were developed using a 1-tissue compartment model and leading-edge delay correction. Results: Estimated baseline perfusion values ranged from 0.02 to 1.95 mL·min-1·cm-3 across organs. Test-retest analysis showed that repeat baseline perfusion measurements were highly correlated (slope, 0.99; Pearson r = 0.96, P < 0.001). For the CPT subject, the largest regional increases were in skeletal muscle (psoas, 142%) and the myocardium (64%). One of the PVD patients showed increased collateral vessel growth in the calf because of a peripheral stenosis. Comorbidities including myocardial infarction, hypothyroidism, and renal failure were correlated with variations in organ-specific perfusion. Conclusion: This pilot study demonstrates the ability to obtain reproducible measurements of total-body perfusion using [11C]-butanol. The methods are sensitive to local perturbations in flow because of physiologic stressors and disease.

Educational video while "waiting-to-be-seen" in a cardiology outpatient clinic promotes opt-in self-consent for biobanking of remnant clinical biospecimens: A randomized-controlled trial.

Objectives: Consenting donors for remnant clinical biospecimen donation is critical for scaling research biorepositories. Opt-in, low-cost, self-consenting for donations that solely relied on clinical staff and printed materials was recently shown to yield ∼30% consent rate. We hypothesized that adding an educational video to this process would improve consent rates. Methods: Randomized patients (by clinic day) in a Cardiology clinic received either printed materials (control) or the same materials plus an educational video on donations (intervention) while waiting to be seen. Engaged patients were surveyed at the clinic checkout for an "opt-in" or "opt-out" response. The decision was documented digitally in the electronic medical record. The primary outcome of this study was the consent rate. Results: Thirty-five clinic days were randomized to intervention (18) or control (17). Three hundred and fifty-five patients were engaged, 217 in the intervention and 158 in the control. No significant demographic differences were noted between treatment groups. Following an intention-to-treat analysis, the rate of opt-in for remnant biospecimen donation was 53% for the intervention and 41% for the control group (p-value = 0.03). This represents a 62% increase in the odds of consenting (OR = 1.62, 95% CI = 1.05-2.5). Conclusion: This is the first randomized trial showing that an educational video is superior to printed materials alone when patients are self-consenting for remnant biospecimen donation. This result adds to the evidence that efficient and effective consenting processes can be integrated into clinical workflows to advance universal consenting in medical research.

Medical Disruptions During Center-Based Cardiac Rehabilitation: A Necessary Appraisal for the Development of Emerging Remote and Virtual Care Models.

PURPOSE: The purpose of this study is to show that with remote and virtual cardiac rehabilitation (CR) care models rapidly emerging, CR core components must be maintained to prioritize safety and effectiveness. Currently, there is a paucity of data on medical disruptions in phase 2 center-based CR (cCR). This study aimed to characterize the frequency and types of unplanned medical disruptions. METHODS: We reviewed 5038 consecutive sessions from 251 patients enrolled in cCR program from October 2018 to September 2021. Quantification of events was normalized to sessions to control for multiple disruptions that occurred to a single patient. A multivariate logistical regression model was used to predict comorbid risk factors for disruptions. RESULTS: Fifty percent of patients experienced one or more disruptions during cCR. Glycemic events (71%) and blood pressure (12%) abnormalities accounted for most of these while symptomatic arrhythmias (8%) and chest pain (7%) were less frequent. Sixty-six percent of events occurred within the first 12 wk. The regression model showed that a diagnosis of diabetes mellitus was the strongest predictor for disruptions (OR = 2.66: 95% CI, 1.57-4.52; P < .0001). CONCLUSIONS: Medical disruptions were frequent during cCR, with glycemic events being most common and occurring early. A diagnosis of diabetes mellitus was a strong independent risk factor for events. This appraisal suggests that patients living with diabetes mellitus, particularly those on insulin, need to be the highest priority for monitoring and planning and suggests that a hybrid care model may be beneficial in this population.

Blanching Defects at the Pressure Points: Observations from Dynamic Total-Body PET/CT Studies.

Total-body PET/CT allows simultaneous acquisition of all the body parts in a single bed position during the radiotracer uptake phase. Dynamic imaging protocols employing total-body PET could demonstrate findings that may not have been previously visualized or described using conventional PET/CT scanners. We examined the characteristics of blanching defects, areas of markedly reduced (partial defect) or absent (complete defect) radiotracer uptake seen at the skin/subcutaneous tissues opposite the bony prominences at pressure points. Methods: In this observational study, 77 participants underwent dynamic total-body PET/CT imaging using 18F-FDG (Group 1, N = 47, 60-min dynamic, arms-down, divided into 3 subgroups according to the injected dose) or 18F-fluciclovine (Group 2, N = 30, 25-min dynamic, arms above the head). 40 out of 47 participants in Group 1 were re-imaged at 90 min after being allowed off the scanning table. Blanching defects, partial or complete, were characterized opposite the bony prominences at 7 pressure points (the skull, scapula, and calcaneus bilaterally, as well as the sacrum). Association of the blanching defects with different clinical and technical characteristics were analyzed using uni- and multi-variate analyses. Results: A total of 124 blanching defects were seen in 68 out of 77 (88%) participants at one or more pressure points. Blanching defects were higher, on average, in Group 2 participants (3.5±1.7) compared to Group 1 (2.1±1.4; P <0.001), but it did not vary within Group 1 for different 18F-FDG dose subgroups. All defects resumed normal pattern on delayed static (90-min) images except for 14 partial defects. No complete blanching defects were seen on the 90-min images. By multivariate analysis, arm positioning above the head was associated with skull defects; scapular and sacral defects were significantly more encountered in men and with lower BMI, while calcaneal defects could not be associated to any factor. Conclusion: Blanching defects opposite the bony pressure points are common on dynamic total-body PET/CT images using different radiopharmaceuticals and injection doses. Their appearance should not be immediately interpreted as an abnormality. The current findings warrant further exploration in a prospective setting and may be utilized to study various mechano-pathologic conditions, such as pressure ulcers.

Multiparametric cardiac18F-FDG PET in humans: pilot comparison of FDG delivery rate with82Rb myocardial blood flow.

Myocardial blood flow (MBF) and flow reserve are usually quantified in the clinic with positron emission tomography (PET) using a perfusion-specific radiotracer (e.g.82Rb-chloride). However, the clinical accessibility of existing perfusion tracers remains limited. Meanwhile,18F-fluorodeoxyglucose (FDG) is a commonly used radiotracer for PET metabolic imaging without similar limitations. In this paper, we explore the potential of18F-FDG for myocardial perfusion imaging by comparing the myocardial FDG delivery rateK1with MBF as determined by dynamic82Rb PET in fourteen human subjects with heart disease. Two sets of FDGK1were derived from one-hour dynamic FDG scans. One was the original FDGK1estimates and the other was the correspondingK1values that were linearly normalized for blood glucose levels. A generalized Renkin-Crone model was used to fit FDGK1with Rb MBF, which then allowed for a nonlinear extraction fraction correction for converting FDGK1to MBF. The linear correlation between FDG-derived MBF and Rb MBF was moderate (r= 0.79) before the glucose normalization and became much improved (r> 0.9) after glucose normalization. The extraction fraction of FDG was also similar to that of Rb-chloride in the myocardium. The results from this pilot study suggest that dynamic cardiac FDG-PET with tracer kinetic modeling has the potential to provide MBF in addition to its conventional use for metabolic imaging.

Multiparametric Cardiac 18F-FDG PET in Humans: Kinetic Model Selection and Identifiability Analysis.

Cardiac 18F-FDG PET has been used in clinics to assess myocardial glucose metabolism. Its ability for imaging myocardial glucose transport, however, has rarely been exploited in clinics. Using the dynamic FDG-PET scans of ten patients with coronary artery disease, we investigate in this paper appropriate dynamic scan and kinetic modeling protocols for efficient quantification of myocardial glucose transport. Three kinetic models and the effect of scan duration were evaluated by using statistical fit quality, assessing the impact on kinetic quantification, and analyzing the practical identifiability. The results show that the kinetic model selection depends on the scan duration. The reversible two-tissue model was needed for a one-hour dynamic scan. The irreversible two-tissue model was optimal for a scan duration of around 10-15 minutes. If the scan duration was shortened to 2-3 minutes, a one-tissue model was the most appropriate. For global quantification of myocardial glucose transport, we demonstrated that an early dynamic scan with a duration of 10-15 minutes and irreversible kinetic modeling was comparable to the full one-hour scan with reversible kinetic modeling. Myocardial glucose transport quantification provides an additional physiological parameter on top of the existing assessment of glucose metabolism and has the potential to enable single tracer multiparametric imaging in the myocardium.

Modeling Clinical Processes to Consent Research Donors of Remnant Biospecimens in an Outpatient Cardiology Clinic.

Introduction: Informed consent for research biospecimen donations is traditionally obtained through a face-to-face interaction with research staff and by signing an Institutional Review Board (IRB)-approved printed form. Electronic signatures (eSign) are routinely used in the electronic medical record (EMR) for the consenting of clinical services after patients review printed documentation. Our goal was to develop an electronic self-consenting workflow that mimicked clinical services. Specifically, we tested a research consent process for the biobanking of remnant clinical samples that relies solely on clinical resources in a busy outpatient practice. Materials and Methods: The Biorepositories Core Resource (BCR) unit initiated a new enterprise-wide biobanking infrastructure for consenting patients, termed Biospecimen Use for Research-Related Investigations and Translational Objectives (BURRITO). BURRITO is modeled after an established clinical process called Terms and Conditions of Service (TACOS). The TACOS requires patients to annually review printed documentation and self-consent electronically for clinical services. BURRITO also requires patients to review printed documentation and self-consent with eSign to opt-in for remnant biospecimen banking, but patients must complete this process only once. We captured eSign for consents directly into the EMR without research staff. Results: Patients reviewed the IRB-approved documents and self-consented during their cardiology clinic visit. At checkout, their participation preferences were electronically documented by clinic staff. During a 6-month period, 123 patients agreed to donate. After a review of process, a second 3-month period identified 202 patients agreeing to donate. BURRITO did not require face-to-face interactions with research staff, used a "no-paper" eSign for consent, and created discrete fields in the clinical EMR of the patient's preference. Conclusions: BURRITO electronically documents informed consent using an EMR functionality and the least amount of clinical and research resources. Our results show promise for developing institutionally adopted processes, which could leverage existing clinical workflows for universal research consenting and scalability.

Genome-wide analysis revealed sex-specific gene expression in asthmatics.

Global gene-expression analysis has shown remarkable difference between males and females in response to exposure to many diseases. Nevertheless, gene expression studies in asthmatics have so far focused on sex-combined analysis, ignoring inherent variabilities between the sexes, which potentially drive disparities in asthma prevalence. The objectives of this study were to identify (1) sex-specific differentially expressed genes (DEGs), (2) genes that show sex-interaction effects and (3) sex-specific pathways and networks enriched in asthma risk. We analyzed 711 males and 689 females and more than 2.8 million transcripts covering 20 000 genes leveraged from five different tissues and cell types (i.e. epithelial, blood, induced sputum, T cells and lymphoblastoids). Using tissue-specific meta-analysis, we identified 439 male- and 297 female-specific DEGs in all cell types, with 32 genes in common. By linking DEGs to the genome-wide association study (GWAS) catalog and the lung and blood eQTL annotation data from GTEx, we identified four male-specific genes (FBXL7, ITPR3 and RAD51B from epithelial tissue and ALOX15 from blood) and one female-specific gene (HLA-DQA1 from epithelial tissue) that are disregulated during asthma. The hypoxia-inducible factor 1 signaling pathway was enriched only in males, and IL-17 and chemokine signaling pathways were enriched in females. The cytokine-cytokine signaling pathway was enriched in both sexes. The presence of sex-specific genes and pathways demonstrates that sex-combined analysis does not identify genes preferentially expressed in each sex in response to diseases. Linking DEG and molecular eQTLs to GWAS catalog represents an important avenue for identifying biologically and clinically relevant genes.

Conspicuity of suspicious breast lesions on contrast enhanced breast CT compared to digital breast tomosynthesis and mammography.

OBJECTIVE: Compare conspicuity of suspicious breast lesions on contrast-enhanced dedicated breast CT (CEbCT), tomosynthesis (DBT) and digital mammography (DM). METHODS: 100 females with BI-RADS 4/5 lesions underwent CEbCT and/or DBT prior to biopsy in this IRB approved, HIPAA compliant study. Two breast radiologists adjudicated lesion conspicuity scores (CS) for each modality independently. Data are shown as mean CS ±standard deviation. Two-sided t-test was used to determine significance between two modalities within each subgroup. Multiple comparisons were controlled by the false-discovery rate set to 5%. RESULTS: 50% of studied lesions were biopsy-confirmed malignancies. Malignant masses were more conspicuous on CEbCT than on DBT or DM (9.7 ±0.5, n = 25; 6.8 ± 3.1, n = 15; 6.7 ± 3.0, n = 27; p < 0.05). Malignant calcifications were equally conspicuous on all three modalities (CEbCT 8.7 ± 0.8, n = 18; DBT 8.5 ± 0.6, n = 15; DM 8.8 ± 0.7, n = 23; p = NS). Benign masses were equally conspicuous on CEbCT (6.6 ± 4.1, n = 22); DBT (6.4 ± 3.8, n = 17); DM (5.9 ± 3.6, n = 24; p = NS). Benign calcifications CS were similar between DBT (8.5 ± 1.0, n = 17) and DM (8.8 ± 0.8, n = 26; p = NS) but less conspicuous on CEbCT (4.0 ± 2.9, n = 25, p < 0.001). 55 females were imaged with all modalities. Results paralleled the entire cohort. 69%(n = 62) of females imaged by CEbCT had dense breasts. Benign/malignant lesion CSs in dense/non-dense categories were 4.8 ± 3.7, n = 33, vs 6.0 ± 3.9, n = 14, p = 0.35; 9.2 ± 0.9, n = 29 vs. 9.4 ± 0.7, n = 14; p = 0.29, respectively. CONCLUSION: Malignant masses are more conspicuous on CEbCT than DM or DBT. Malignant microcalcifications are equally conspicuous on all three modalities. Benign calcifications remain better visualized by DM and DBT than with CEbCT. We observed no differences in benign masses on all modalities. CS of both benign and malignant lesions were independent of breast density. ADVANCES IN KNOWLEDGE: CEbCT is a promising diagnostic imaging modality for suspicious breast lesions.

The Doctor-Patient Relationship With Artificial Intelligence.

OBJECTIVE: The doctor-patient relationship has been evolving from benevolent paternalism to a more patient-centered relationship in the modern era. Although artificial intelligence (AI) has the potential to improve nearly every aspect of health care, many physicians are skeptical about integrating AI into their current medical practice. The purpose of this article is to explore what AI means for the doctor-patient relationship and for breast imaging radiologists. CONCLUSION: The promise of AI is its potential to release physicians from tasks that are better performed by automation. AI may enhance our diagnostic accuracy to the point that we are able to refocus on the art of the doctor-patient relationship.

Engaging heart failure patients from a clinical data research network: A survey on willingness to participate in different types of research.

The willing participation of patients in clinical research is a critical element in national efforts to collect health data for precision medicine and large cohort studies. However, recruiting patients is challenging. Clinical data research networks (CDRN) have primarily been used for observational studies, but may be able to enhance recruitment efforts. We need a better understanding of patient motivation and preferences for research participation and their interest in different types of research activities, particularly among those who are already represented in CDRNs. We surveyed a heart failure patient cohort constructed from EHRs in a CDRN to assess research participation. Results showed that CDRN recruitment is feasible. Respondents were most interested in completing a one-time survey and giving a blood sample one time. They were least interested in a study about weight control that require surgery. We found statistically significant associations between race and research activity interests.

Implementation of High-Sensitivity Cardiac Troponin: Challenges From the International Experience.

OBJECTIVE: Implementation of the newly approved high-sensitivity cardiac troponin (hs-cTn) in the United States presents a challenge for clinical practice. Sex-specific cutoffs, clinical protocols, and workflows will likely require modifications before implementation. METHODS: We conducted a cross-sectional survey of international physicians and laboratorians already utilizing hs-cTn for the evaluation of acute myocardial infarction. RESULTS: Twenty-two of 54 (41%) eligible participants completed the survey, representing 9 countries and 18 hospitals. All reported successful hs-cTn implementation and diagnostic utility (mean 8.6 + 1.2 out of 10 for best implementation). The major perceived benefit was more rapid evaluation of acute myocardial infarction (14/19, 74%), and the most frequently cited limitation was an increase in the number of measurable hs-cTn values that required further evaluation (8/18, 44%). Institutions using the hs-cTnI assay favored sex-specific cutoffs (5/6, 83%), whereas institutions employing the hs-cTnT assay favored a combined cutoff (12/12, 100%). Timing of serial hs-cTn measurements varied, with 0-3 hours (8/17, 47%) most frequent, followed by 0-2 hours (4/17, 24%), 0-1 hour (3/17, 18%), and other (2/17, 12%). CONCLUSIONS: Our survey of hs-cTn implementation at international institutions reveals satisfaction with new assays but reflects important variations in clinical practice. The use of sex-specific vs. combined cutoffs and timing of serial hs-cTn measurements varies across institutions and are subjects that United States centers must define without consensus from international practices.

Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles.

PURPOSE: The delivery of transgenes into human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiPSC-CMs) represents an important tool in cardiac regeneration with potential for clinical applications. Gene transfection is more difficult, however, for hiPSCs and hiPSC-CMs than for somatic cells. Despite improvements in transfection and transduction, the efficiency, cytotoxicity, safety, and cost of these methods remain unsatisfactory. The objective of this study is to examine gene transfection in hiPSCs and hiPSC-CMs using magnetic nanoparticles (NPs). METHODS: Magnetic NPs are unique transfection reagents that form complexes with nucleic acids by ionic interaction. The particles, loaded with nucleic acids, can be guided by a magnetic field to allow their concentration onto the surface of the cell membrane. Subsequent uptake of the loaded particles by the cells allows for high efficiency transfection of the cells with nucleic acids. We developed a new method using magnetic NPs to transfect hiPSCs and hiPSC-CMs. HiPSCs and hiPSC-CMs were cultured and analyzed using confocal microscopy, flow cytometry, and patch clamp recordings to quantify the transfection efficiency and cellular function. RESULTS: We compared the transfection efficiency of hiPSCs with that of human embryonic kidney (HEK 293) cells. We observed that the average efficiency in hiPSCs was 43%±2% compared to 62%±4% in HEK 293 cells. Further analysis of the transfected hiPSCs showed that the differentiation of hiPSCs to hiPSC-CMs was not altered by NPs. Finally, robust transfection of hiPSC-CMs with an efficiency of 18%±2% was obtained. CONCLUSION: The difficult-to-transfect hiPSCs and hiPSC-CMs were efficiently transfected using magnetic NPs. Our study offers a novel approach for transfection of hiPSCs and hiPSC-CMs without the need for viral vector generation.

Impact of the California Breast Density Law on Screening Breast MR Utilization, Provider Ordering Practices, and Patient Demographics.

PURPOSE: To assess the impact of California's Breast Density Law (BDL) on MRI utilization and clinician ordering practices. MATERIALS AND METHODS: Our institutional review board approved this study that retrospectively compared the ordering pattern for screening breast MRI examinations in the 30-month period before and after the BDL was enacted. Examinations were subcategorized into those with breast density mentioned as an examination indication. Patients were classified into (1) high risk; (2) above average risk, defined but not quantified; and (3) undefined or average risk. χ2 test or Fisher's exact test was used to compare MRI utilization, use of breast density as an indication, patient demographics, and provider characteristics. RESULTS: Screening MRI examinations with breast density as the indication increased from 8.5% (32 of 376) to 21.1% (136 of 646, P < .0001) after BDL. When high-risk patients were excluded, the increase was from 8% to 17.2% (P < .0001). Patient demographics before and after BDL were, by race: white 71.8% versus 71.2%; Asian 6.4% versus 10.5%; black 3.7% versus 3.1%; American Indian 0.3% versus 1.4%; Native Hawaiian or Pacific Islander 1.6% versus 1.7%; by ethnicity: Hispanic or Latino 10.6% versus 7.9%. Before and after BDL, predominantly female providers (81.4% and 77.4%, P = not significant [NS]) and specialists (62.5% and 63.5%, P = NS) ordered the majority of breast MRI examinations compared with males (18.6% and 22.6%, P = NS). CONCLUSION: Screening breast MRI utilization for non-high-risk women more than doubled after the California BDL went into effect. BDL has had an impact on MRI utilization, and its clinical value for changing outcomes deserves further study.

Aspirin and clopidogrel high on-treatment platelet reactivity and genetic predictors in peripheral arterial disease.

OBJECTIVES: Our aims were to examine the prevalence and genetic predictors of aspirin and clopidogrel high on-treatment platelet reactivity (HoTPR), and associated adverse cardiovascular outcomes in patients with peripheral arterial disease (PAD). BACKGROUND: The association of aspirin and clopidogrel HoTPR with outcomes in PAD remains unclear. METHODS: This is a prospective cohort study of patients with angiographically documented PAD involving carotid and lower extremity arteries. Aspirin and clopidogrel HoTPR (using the VerifyNow Assay) and associated genetic predictors were compared to clinical outcomes. The primary end-point was a composite of major adverse cardiovascular events: all-cause mortality, myocardial infarction, stroke, target vessel revascularization (TVR) and limb-loss in patients who underwent extremity intervention. RESULTS: The study was stopped prematurely due to slow patient enrolment. Of 195 patients enrolled, the primary analysis was performed in 154 patients taking both drugs. Aspirin HoTPR was present in 31 (20%) and clopidogrel HoTPR in 76 (49%) patients. There was a trend toward more primary composite outcome events with PRU ≥ 235 (52% freedom-from-event rate vs. 70% for PRU < 235; P = 0.09). TVR was higher in those with PRU ≥ 235 (20 vs. 6%, unadjusted P = 0.02). There was no association between aspirin HoTPR and combined outcomes. Single nucleotide polymorphisms in serum paraoxonase/arylesterase 1 (PON1) gene was associated with aspirin HoTPR (P = 0.005) while SNP in phospholipase A2, group III (PLA2G3) gene was associated with clopidogrel HoTPR (P = 0.002). CONCLUSION: Clopidogrel HoTPR was significantly associated with TVR, while aspirin HoTPR was not associated with adverse clinical outcomes in patients with PAD.

A method to increase reproducibility in adult ventricular myocyte sizing and flow cytometry: Avoiding cell size bias in single cell preparations.

RATIONALE: Flow cytometry (FCM) of ventricular myocytes (VMs) is an emerging technology in adult cardiac research that is challenged by the wide variety of VM shapes and sizes. Cellular variability and cytometer flow cell size can affect cytometer performance. These two factors of variance limit assay validity and reproducibility across laboratories. Washing and filtering of ventricular cells in suspension are routinely done to prevent cell clumping and minimize data variability without the appropriate standardization. We hypothesize that washing and filtering arbitrarily biases towards sampling smaller VMs than what actually exist in the adult heart. OBJECTIVE: To determine the impact of washing and filtering on adult ventricular cells for cell sizing and FCM. METHODS AND RESULTS: Left ventricular cardiac cells in single-cell suspension were harvested from New Zealand White rabbits and fixed prior to analysis. Each ventricular sample was aliquoted before washing or filtering through a 40, 70, 100 or 200µm mesh. The outcomes of the study are VM volume by Coulter Multisizer and light-scatter signatures by FCM. Data are presented as mean±SD. Myocyte volumes without washing or filtering (NF) served as the "gold standard" within the sample and ranged from 11,017 to 46,926µm3. Filtering each animal sample through a 200µm mesh caused no variation in the post-filtration volume (1.01+0.01 fold vs. NF, n = 4 rabbits, p = 0.999) with an intra-assay coefficient of variation (%CV) of <5% for all 4 samples. Filtering each sample through a 40, 70 or 100µm mesh invariably reduced the post-filtration volume by 41±10%, 9.0±0.8% and 8.8±0.8% respectively (n = 4 rabbits, p<0.0001), and increased the %CV (18% to 1.3%). The high light-scatter signature by FCM, a simple parameter for the identification of ventricular myocytes, was measured after washing and filtering. Washing discarded VMs and filtering cells through a 40 or 100µm mesh reduced larger VM by 46% or 11% respectively (n = 6 from 2 rabbits, p<0.001). CONCLUSION: Washing and filtering VM suspensions through meshes 100µm or less biases myocyte volumes to smaller sizes, excludes larger cells, and increases VM variability. These findings indicate that validity and reproducibility across laboratories can be compromised unless cell preparation is standardized. We propose no wash prior to fixation and a 200µm mesh for filtrations to provide a reproducible standard for VM studies using FCM.