bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimal Utilization of Ports’ Free‐of‐Charge Times in One Distribution Center and Multiple Ports Inventory Systems
Authors: Li Zhengyi
Published: February 2019

In this paper, we consider a distribution system consisting of one distribution center (DC), a set of ports, and a set of retailers, in which the product is distributed to the retailers from the DC through the ports by the water transport, and study inventory management for the distribution system with considering the effect of the free storage periods provided by the ports. Inventory management for the distribution system is to determine the order intervals of the DC and the retailers while minimizing the inventory ordering and holding costs. Focusing on stationary and integer‐ratio policies, we formulate this inventory management problem as an optimization problem with a convex objective function and a set of integer‐ratio constraints and present O(Nlog⁡N) time algorithm to solve the relaxed problem (relaxing the integer‐ratio constraints) to optimality, where N is the number of the retailers. We prove that the relaxed problem provides a lower bound on average cost for all the feasible policies (containing dynamic policies) for this inventory management problem. By using the optimal solution of the relaxed problem, we build a stationary integer‐ratio policy (a power‐of‐two policy) for this inventory management problem and prove that the power‐of‐two policy can approximate the optimal inventory policy to 83% accuracy.
Toxicity of DON on GPx1-Overexpressed or Knockdown Porcine Splenic Lymphocytes In Vitro and Protective Effects of Sodium Selenite
Authors: Ren Zhihua , Chen Changhao , Fan Yu , Chen Chaoxi , He Hongyi , Wang Xuemei , Zhang Zhuo , Zuo Zhicai , Peng Guangneng , Hu Yanchun , Xu Zhiwen , Tao Siyi , Mao Xinru , Deng Junliang
Published: February 2019

Deoxynivalenol (DON) is a common contaminant of grain worldwide and is often detected in the human diet and animal feed. Selenium is an essential trace element in animals. It has many biological functions. The role of selenium in the body is mainly orchestrated by selenoprotein. Glutathione peroxidase (GPx) also exists widely in the body and has attracted much attention due to its high antioxidant capacity. In order to explore the effect of the GPx1 gene on toxicity of DON, in this study, we overexpressed or knockdown GPx1 in porcine splenic lymphocytes, then added different concentrations of DON (0.1025, 0.205, 0.41, and 0.82 μg/mL) and sodium selenite (2 μmol/L) to the culture system. Using various techniques, we detected antioxidant function, free radical content, cell apoptosis, and methylation-related gene expression to explore the effect of GPx1 expression on DON-induced cell damage. We also explored whether selenium can antagonize the toxicity of DON in these two cell models and revealed the protective effect of sodium selenite on DON-induced cell damage in GPx1-overexpressing or knockdown splenic lymphocytes. Finally, our findings revealed the following: (1) GPx1 can regulate the antioxidant capacity, apoptosis rate, and expression of DNA methylation-related genes in pig splenic lymphocytes. (2) Na2SeO3 (2 μmol/L) can regulate the antioxidant capacity, apoptosis rate, and expression of DNA methylation-related genes in pig splenic lymphocytes, and this effect is more significant in GPx1-overexpressing cells than in GPx1-knockdown cells. (3) DON can cause oxidative damage, apoptosis, and methylation injury in GPx1-overexpressing or knockdown pig splenic lymphocytes in a concentration-dependent manner. (4) Na2SeO3 (2 μmol/L) can antagonize the toxic effect of DON on GPx1-overexpressing or knockdown pig splenic lymphocytes. Our findings may have important implications for food/feed safety, human health, and environmental protection.
Colorectal Cancer Characteristics and Outcomes after Solid Organ Transplantation
Authors: Merchea Amit , Shahjehan Faisal , Croome Kristopher P. , Cochuyt Jordan J. , Li Zhuo , Colibaseanu Dorin T. , Kasi Pashtoon Murtaza
Published: February 2019

Background. Individuals after solid organ transplant may develop secondary malignancies. In our clinical practice, we noted an increasing number of individuals who developed colorectal cancers after solid organ transplantation. The primary aim of this study was to describe the characteristics and outcomes of the patients who developed colorectal cancer after solid organ transplant. Materials and Methods. Data was gathered and merged from several registries at Mayo Clinic to identify all patients who received a diagnosis of colon or rectal cancer and solid organ transplant. Continuous variables were summarized as mean (standard deviation) and median (range), while categorical variables were reported as frequency (percentage). Time to colorectal cancer after transplant and overall survival after cancer diagnosis were estimated using Kaplan-Meier method. Results. Initially, 115 colorectal cancer patients who also had a transplant were identified. The diagnosis of colorectal cancer was noted after solid organ transplant in 63 patients. The mean age at transplant was 57 years. Majority had received a kidney transplant (44.4%) followed by liver (36.5%). The median time to develop colorectal cancer was 59.3 months (range: 4.4-251.4 months). 15 (24.6%) were stage 4 at diagnosis and 13 (21.3%) had stage 3 colorectal cancer. Median overall survival was 30.8 months; 5-, 10- and 15-year survival were noted to be 42.5%, 17.9%, and 7.5%, respectively. None of the stage 4 patients were alive at 5 years; 5-year survival rate for stage 1, 2, and 3 patients was 77%, 50%, and 42%, respectively. Conclusions. Our study reports on one of the largest cohorts of patients of colorectal cancer that developed the cancer after solid organ transplant. Survival is extremely poor for advanced cases. However, long-term survivors are noted who developed the cancer at a relatively early stage. Colorectal screening recommendations may need to be revised for patients after solid organ transplant.
Histobiomechanical Remodeling of the Cervix during Pregnancy: Proposed Framework
Authors: Torres Jorge , Faris Inas , Callejas Antonio
Published: February 2019

Pregnancy involves a gradual change in tissue consistency, where, as gestational age increases a drop in stiffness is noticeable. The extracellular matrix (ECM) of the cervix whose behavior is controlled by the collagen structure is reconfigured, due to hormonal and protein related factors that degraded it. There is an increase in cellular and water content. The collagen maintains a stable percentage, although the synthesis‐solubility interaction causes a transformation in the network increasing its diameter and waviness, which destabilizes the mechanical integrity. Further understanding of the remodeling of the cervix becomes a key element in the progress against preterm birth. However, since the woman is in a very delicate state, the available information is scattered. The aim is to provide a framework where the variation of the histological properties is linked with the individual biomechanical evolution of the constituents; therefore a multiscale approach to the problem can be made. The results of different authors were reviewed to obtain values of each component depending on the gestational age, and then they were combined with the mechanical development of each variable obtained from the outputs of numerical simulations of another author. An additive function is proposed, in which the mechanical contribution of each constituent is added to a reference value established for a nonpregnant state. Results show how biochemistry models the mechanical behavior of the tissue through the histology and morphology of the ECM.
Modified Mechanistic Model Based on Gaussian Process Adjusting Technique for Cutting Force Prediction in Micro‐End Milling
Authors: Liao Xiaoping , Zhang Zhenkun , Chen Kai , Li Kang , Ma Junyan , Lu Juan
Published: February 2019

Micro‐end milling is in common use of machining micro‐ and mesoscale products and is superior to other micro‐machining processes in the manufacture of complex structures. Cutting force is the most direct factor reflecting the processing state, the change of which is related to the workpiece surface quality, tool wear and machine vibration, and so on, which indicates that it is important to analyze and predict cutting forces during machining process. In such problems, mechanistic models are frequently used for predicting machining forces and studying the effects of various process variables. However, these mechanistic models are derived based on various engineering assumptions and approximations (such as the slip‐line field theory). As a result, the mechanistic models are generally less accurate. To accurately predict cutting forces, the paper proposes two modified mechanistic models, modified mechanistic models I and II. The modified mechanistic models are the integration of mathematical model based on Gaussian process (GP) adjustment model and mechanical model. Two different models have been validated on micro‐end‐milling experimental measurement. The mean absolute percentage errors of models I and II are 7.76% and 6.73%, respectively, while the original mechanistic model’s is 15.14%. It is obvious that the modified models are in better agreement with experiment. And model II performs better between the two modified mechanistic models.
Finding Plausible Optimal Solutions in Engineering Problems Using an Adaptive Genetic Algorithm
Authors: Kilinc Muslum , Caicedo Juan M.
Published: February 2019

In engineering, optimization applications are commonly used to solve various problems. As widely known, solution of an engineering problem does not have a unique result; moreover, the solution of a unique problem may totally differ from one engineer to another. On the other hand, one of the most commonly used engineering optimization methods is genetic algorithm that leads us to only one global optimum. As to mention, engineering problems can conclude in different results from the point of different engineers’ views. In this study, a modified genetic algorithm named multi‐solution genetic algorithm (MsGA) based on clustering and section approaches is presented to identify alternative solutions for an engineering problem. MsGA can identify local optima points along with global optimum and can find numerous solution alternatives. The reliability of MsGA was tested by using a Gaussian and trigonometric function. After testing, MsGA was applied to a truss optimization problem as an example of an engineering optimization problem. The result obtained shows that MsGA is successful at finding multiple plausible solutions to an engineering optima problem.
Hepatoprotective Potential ofSargassum muticumagainst STZ-Induced Diabetic Liver Damage in Wistar Rats by Inhibiting Cytokines and the Apoptosis Pathway
Authors: Safhi Mohammed M. , Alam Mohammad Firoz , Sivakumar Sivagurunathan Moni , Anwer Tarique
Published: February 2019

Liver inflammation and necrosis are the foremost problems interlinked with diabetes mellitus (DM). The methanolic extract ofSargassum muticum(MESM) plays a hepatoprotective role in streptozotocin- (STZ-) induced hepatic injury. In this study, STZ exposure induced diabetes that augmented hepatic damage, which was reflected in serum enzyme markers, the cytokine network, and caspase-3 and caspase-9 levels in Group 2. Exposure to the MESM tremendously modulated the levels of hepatic enzyme markers ALP, ACP, ALT, and AST in Groups 3 and 4. The cytokine network was well regulated by suppressing the release of cytokines, and the levels of caspase-3 and caspase-9 were also reduced in Groups 3 and 4. The present study suggests that MESM treatment at 200 and 500 mg protected the liver and also minimizes the glucose level. Thus, the MESM plays a key role in rejuvenating the liver and can modulate diabetes’s pathogenic effect by reducing the glucose level.
Consensus of Multiagent System in the Sense of Curvature and Torsion
Authors: Mou Jinping
Published: February 2019

In this paper, the consensus problem is investigated for a distributed multiagent system (MAS), where the consensus is characterized by curvature function and torsion function. According to the Frenet‐Serret formulas, a distributed consensus protocol is designed for the tangent, normal, and binormal unit vectors of trajectory of each agent, and then it gets a closed‐loop system. Based on the Lyapunov function, several sufficient conditions for consensus are derived for the closed‐loop system. Also the consensus problem of multiagent system on a surface is studied. Finally, the numerical examples show the reliability of the proposed methods.
Optimization of Energy Recovery Efficiency for Parallel Hydraulic Hybrid Power Systems Based on Dynamic Programming
Authors: Ning Xiaobin , Shangguan Jiarong , Xiao Yong , Fu Zhijun , Xu Gaolun , He Anqing , Li Bin
Published: February 2019

In this paper, an optimization algorithm of energy recovery efficiency is proposed for parallel hydraulic hybrid systems (PHHS) using dynamic programming (DP). Global optimal solution of pump displacement and transmission ratio under the known urban drive cycles is obtained by using the DP approach, where the total amount of energy recovery is defined as the cost function, and the pump displacement and the transmission ratio of the torque coupler are defined as the deciding variables. Two major steps are involved in verifying the proposed approach. Firstly, a PHHS Simulink model is accurately obtained by repeated comparison with the bench test. Subsequently, we derive a parallel hydraulic hybrid vehicle (PHHV) from adding a hydraulic hybrid system to an electric vehicle in ADVISOR (advanced vehicle simulator). This vehicle is used to validate the effectiveness of the proposed method in energy recovery efficiency.
Loneliness Shapes the Relationship between Information and Communications Technology Use and Psychological Adjustment among Older Adults
Authors: Fang Yang , Chau Anson K.C. , Fung Helene H. , Woo Jean
Published: February 2019

Background: Given findings that generally support the benefits of information and communication technology (ICT) for older adults’ psychosocial adjustment, one might surmise that lonely older adults, who have a stronger need for psychological support, would reap more psychosocial benefits from ICT use. However, scant research has examined this view, much less the likelihood that ICT use might worsen the psychological well-being of lonely older adults, as has been shown to be the case in younger adults. Objective: To examine whether the association between ICT use and psychological adjustment (i.e., psychological distress and sense of community [SOC]) among older adults depends on their loneliness levels. Methods: A representative sample of 738 Hong Kong SAR Chinese older adults aged 60 years or older (56% female) was interviewed in 2017 on loneliness, frequency of ICT use (i.e., Internet and smart devices), psychological distress (6-item Kessler scale; K6), and SOC. Results: Regression analyses showed that loneliness significantly moderated the relationship between ICT use frequency and psychological adjustment (psychological distress and SOC); more frequent ICT use was associated with more psychological distress and less SOC, with higher levels of loneliness. Conclusion: These findings suggest that ICT use may be a mixed blessing for older adults, i.e., using more ICT might predict worse psychological adjustment among lonelier older adults. Efforts that promote ICT use among older adults should take their loneliness levels into account.
Multidisciplinary Tumor Board Recommendations for Oligometastatic Malignancies: A Prospective Single-Center Analysis
Authors: Galata Christian , Wimmer Elisabeth , Kasper Bernd , Wenz Frederik , Reißfelder Christoph , Jakob Jens
Published: February 2019

Background: The treatment of oligometastatic disease is challenging and few data exist to guide treatment decisions. The objective of this study was to improve the data on the prevalence and treatment of patients with oligometastatic disease. Methods: We conducted a prospective single-center analysis that included all consecutive patients discussed in multidisciplinary tumor boards (MDT) between February and July 2017. Patients with oligometastatic disease were identified and treatment strategies were evaluated. Results: 1,673 patients were included in this study, 609 (36.4%) presented with metastatic disease, 151 (9%) had oligometastatic disease. Common metastatic sites were brain, liver, and lung. Lung cancer patients were the largest cohort (20.5%) among all patients with oligometastatic disease compared with other tumor entities. The majority of oligometastatic patients (68.9%) received local treatment with or without additional chemotherapy, 17.9% were recommended systemic therapy alone. MDT recommended watchful waiting for 4.6% of the patients. Conclusion: Patients with oligometastatic disease represent a considerable proportion of all patients in MDT. In this study, 68.9% of patients with oligometastatic disease received regional treatment. This shows a possible treatment shift from palliative to potentially curative intent. These data may be used to design prospective clinical trials to optimize the treatment of oligometastatic disease.
The Association between Long- and Intra-Dialytic Blood Pressure Variability with All-Cause Mortality in Hemodialysis Patients
Authors: Liao Ruoxi , Li Jiameng , Lin Liping , Sun Si , Wang Liya , Xiong Yuqin , Li Yupei , Han Mei , Liu Qiang , Su Baihai
Published: February 2019

Background: Blood pressure variability (BPV) is a potential prognostic predictor for all-cause mortality. Objectives: We conducted a retrospective cohort study to compare the prognostic value of long-term BPV with intra-dialytic BPV in hemodialysis (HD) patients. Materials and Methods: We included 611 HD patients and collected their baseline blood pressure (BP) measurements for 1 year and monitored them for 40 months. Long-term BPV was assessed by pre-dialysis BP SD and pre-dialysis absolute BP residual metric. Intra-dialytic BPV was assessed by intra-dialytic BP average real variability and intra-dialytic absolute BP residual. Results: Long-term systolic BPV showed a weak correlation with mean BP, but a stronger correlation with intra-dialytic BPV. High long-term systolic blood pressure (SBP) SD and long-term SBP residual metrics were associated with high all-cause mortality (p = 0.0084 and 0.0056, respectively), while no such association was found for intra-dialytic BPV or diastolic BPV. According to receiver operating characteristic curve with mortality as dependent variable, long-term SBP residual metric showed the strongest prognostic ability (area under curve [AUC] 0.679, p = 0.0006), which was even stronger in patients with BP ≥140/90 mm Hg (AUC 0.713, p = 0.0004). After completely adjusting for confounders, long-term SBP residual metric remained significantly associated with all-cause mortality (hazard ratio 1.628 per quartile; 95% CI 1.086–2.441). Conclusions: Our results suggest long-term SBP residual metric to be a better predictor of all-cause mortality in HD patients, which could be used as an additional target for BP management.
Antidiabetic Medications and the Risk of Endometrial Cancer in Patients
Authors: Tian Jing , Liang Yuan , Qu Pengpeng
Published: February 2019

Background: Recent epidemiological studies suggest that antidiabetic medications (ADMs) may reduce the risks of cancer and cancer mortality among patients with type 2 diabetes. However, data on its effect on endometrial cancer (EC) are unclear. In this systematic review and meta-analysis, we evaluated the association between metformin, sulfonylureas (SUs), insulin, and the risk of EC in patients with diabetes mellitus (DM). Methods: Studies were identified from PubMed and EMBASE database through October 2017. Observational studies reporting relative risks OR or provided data for their estimation were selected. Summary OR estimates with 95% CIs were estimated using the random-effects model. Results: Six studies (2 retrospective cohorts and 4 case-control trials) reported 4,739 cases of EC in 510,344 patients with DM. Meta-analysis of observational studies showed no significant association between metformin (n = 6 studies; unadjusted OR 1.15; 95% CI 0.70–1.88; p = 0.59), SUs (unadjusted OR 1.11, 95% CI 0.94–1.32, p = 0.23), or insulin (n = 3 studies; unadjusted OR 1.15; 95% CI 0.93–1.40; p = 0.19) use and risk of developing EC. However, when we do heterogeneity and sensitivity analysis, the use of metformin was associated with an increased incidence of endometrial EC compared to no metformin use (unadjusted OR 1.29; 95% CI 1.16–1.44; p < 0.01). Conclusion: Our data indicate that metformin, SUs, and insulin did not meaningfully affect the risk of EC. Further investigation is warranted.
Novel Association between Flavin-Containing Monooxygenase 3 Gene Polymorphism and Antithyroid Drug-Induced Agranulocytosis in the Han Population
Authors: He Ya-Yi , Hasan Abdulbaqi Mohamed Esmail , Zhang Qian , Li Shao-Qing , Yang Jing-Si , Yan Chun-Xia , Chen Pu , Liu Yan , Nadeem Asif , Zhang Bao
Published: February 2019

Some effective antithyroid drugs (ATDs) have been widely used for patients with Graves’ disease (GD) but are associated with ATD-induced agranulocytosis. We selected 29 ATD-induced agranulocytosis patients, 44 ATD-induced neutropenia patients, and 140 GD controls among the Chinese Han population who were recruited at the First Affiliated Hospital of Xi’an Jiao Tong University. We assessed their response to ATDs treatment by performing genotyping for a candidate gene association study of samples from patients receiving treatment. Human flavin-containing monooxygenase 3 (FMO3), which is the major hepatic enzyme involved in the production of N-oxide of trimethylamine, catalyzes the oxygenation of a variety of drug compounds. Six single SNP, genotype, haplotype (HAP), and association analyses of the FMO3 gene with ATD-induced agranulocytosis/neutropenia under different models (i.e., additive, dominant, and recessive models) were performed. Rs1736557, which caused an amino acid variation V257M, showed a strong association between ATD-induced agranulocytosis and GD controls after Bonferroni correction (p = 0.011, OR 2.301, 95% CI 1.201–4.409). The presence of HAP 3 (HAP3) in the FMO3 gene HAP was statistically associated with ATD-induced agranulocytosis (p = 0.038, permutation p value). Our findings indicate that genetic variations in the FMO3 gene are associated with the response to ATDs maintenance treatment in ATD-induced agranulocytosis patients of ­Chinese Han population.
Serum miR-192 Is Related to Tubulointerstitial Lesion and Short-Term Disease Progression in IgA Nephropathy
Authors: Fan Qichen , Lu Renhua , Zhu Mingli , Yan Yucheng , Guo Xiangjiang , Qian Yingying , Zhang Lulu , Dai Huili , Ni Zhaohui , Gu Leyi
Published: February 2019

Background/Aims: Clinical manifestation and renal histology serve as the current “gold standard” for evaluation of renal lesions in IgA nephropathy. MiR-192 is regarded as a potential noninvasive biomarker for kidney disease. We sought to elucidate a relationship between intrarenal, serum, and urinary exosomal miR-192 with renal lesions and disease progression in IgA nephropathy. Methods: Serum and urinary exosomal miR-192 were detected and correlated with clinical and pathological parameters from consecutive IgA nephropathy patients (n = 50) and healthy control patients (n = 25). Patients then received a follow-up of 24 months after biopsy. Disease progression was defined as a 40% reduction in estimated glomerular filtration rate. Expression of miR-192 was quantified by Taqman RT-PCR. Intrarenal ­miR-192 was detected in 8 IgA nephropathy patients and 4 matched patients receiving kidney nephrectomy as controls via in situ hybridization. TGF-β1 and E-cadherin expression in renal tissue was evaluated using immunohistochemistry. Results: Intrarenal miR-192 was correlated with serum miR-192 (r = 0.690, p = 0.013). Both intrarenal and serum miR-192 decreased in IgA nephropathy patients and were correlated with estimated glomerular filtration ratio. Patients with lower intrarenal and serum miR-192 levels had a higher interstitial fibrosis and tubular atrophy score, more severe lesions in tubular atrophy, and interstitial inflammation. Renal E-cadherin expression was correlated (r = 0.484, p = 0.004) and TGF-β1 was inversely correlated (r = –0.527, p < 0.001) with serum miR-192 in IgA. Patients with higher serum miR-192 had a lower disease progression rate over the course of 2 years (0 vs. 16%, p = 0.028). No correlation was found in urinary exosomal miR-192 with the clinical and pathological parameters mentioned earlier. Conclusions: Lower serum miR-192 in IgA nephropathy patients indicates lower intrarenal miR-192 expression, more severe tubular atrophy, interstitial inflammation, and fibrotic tendency (with higher TGF-β and E-cadherin in renal miR-192). IgA nephropathy patients with higher serum miR-192 are less likely to have renal function decline over the course of 2 years.
Novel Antiatherosclerotic Therapies
Authors: Libby Peter , Everett Brendan M.
Published: February 2019

Many measures can control lipid risk factors for atherosclerosis. Yet, even with excellent control of dyslipidemia, other sources of risk remain. Hence, we must look beyond lipids to address residual risk. Lifestyle measures should form the foundation of cardiovascular risk control. Many pharmacological interventions targeting oxidation have proven disappointing. A large program tested inhibition of a LpPLA
2
(lipoprotein-associated phospholipase A
2
), culminating in 2 large-scale clinical trials that did not meet their primary end points. A variety of antioxidants have not shown benefit in clinical trials. Numerous laboratory and clinical studies have inculpated inflammatory pathways in the pathogenesis of atherosclerotic events. The p38 MAPK (mitogen-activated protein kinase) inhibitor losmapimod and an inhibitor of a leukocyte adhesion molecule, P-selectin, did not alter adverse events in trials. Low-dose methotrexate, despite the promising observational studies, did not lower biomarkers of inflammation or alter cardiovascular outcomes in the CIRT (cardiovascular inflammation reduction trial). Four large-scale investigations underway will determine colchicine’s ability to reduce recurrent events in secondary prevention. The CANTOS (Canakinumab Anti-inflammatory Thrombosis Outcomes Study) showed that an antibody that neutralizes IL (interleukin)-1β can reduce recurrent cardiovascular events in secondary prevention. The success of CANTOS points to the pathway that leads from the NLRP3 (NOD-like receptor family, pyrin domain-containing protein 3) inflammasome through IL-1β to IL-6 as an attractive target for further study and clinical development beyond lipid therapies to address the unacceptable burden of risk that remains despite our best current care in secondary prevention.
DKK3 (Dikkopf-3) Transdifferentiates Fibroblasts Into Functional Endothelial Cells—Brief Report
Authors: Chen Ting , Karamariti Eirini , Hong Xuechong , Deng Jiacheng , Wu Yutao , Gu Wenduo , Simpson Russell , Wong Mei Mei , Yu Baoqi , Hu Yanhua , Qu Aijuan , Xu Qingbo , Zhang Li
Published: February 2019

Objective—

To determine the role of a cytokine-like protein DKK3 (dikkopf-3) in directly transdifferentiating fibroblasts into endothelial cells (ECs) and the underlying mechanisms.

Approach and Results—

DKK3 overexpression in human fibroblasts under defined conditions for 4 days led to a notable change in cell morphology and progenitor gene expression. It was revealed that these cells went through mesenchymal-to-epithelial transition and subsequently expressed KDR (kinase insert domain receptor) at high levels. Further culture in EC defined media led to differentiation of these progenitors into functional ECs capable of angiogenesis both in vitro and in vivo, which was regulated by the VEGF (vascular endothelial growth factor)/miR (microRNA)-125a-5p/Stat3 (signal transducer and activator of transcription factor 3) axis. More importantly, fibroblast-derived ECs showed the ability to form a patent endothelium-like monolayer in tissue-engineered vascular grafts ex vivo.

Conclusions—

These data demonstrate that DKK3 is capable of directly differentiating human fibroblasts to functional ECs under defined media and provides a novel potential strategy for endothelial regeneration.
Endothelial TFEB (Transcription Factor EB) Restrains IKK (IκB Kinase)-p65 Pathway to Attenuate Vascular Inflammation in Diabetic
db/db
Mice
Authors: Song Wencong , Zhang Cheng-Lin , Gou Lingshan , He Lei , Gong Yao-Yu , Qu Dan , Zhao Lei , Jin Nana , Chan Ting Fung , Wang Li , Tian Xiao Yu , Luo Jiang-Yun , Huang Yu
Published: February 2019

Objective—

TFEB (transcription factor EB) was recently reported to be induced by atheroprotective laminar flow and play an anti-atherosclerotic role by inhibiting inflammation in endothelial cells (ECs). This study aims to investigate whether TFEB regulates endothelial inflammation in diabetic
db/db
mice and the molecular mechanisms involved.

Approach and Results—

Endothelial denudation shows that TFEB is mainly expressed in ECs in mouse aortas. Western blotting shows TFEB total protein level decreases whereas the p-TFEB S142 (phosphorylated form of TFEB) increases in
db/db
mouse aortas, suggesting a decreased TFEB activity. Adenoviral TFEB overexpression reduces endothelial inflammation as evidenced by decreased expression of vascular inflammatory markers in
db/db
mouse aortas, and reduced expression of a wide range of adhesion molecules and chemokines in human umbilical vein ECs. Monocyte attachment assay shows TFEB suppresses monocyte adhesion to human umbilical vein ECs. RNA sequencing of TFEB-overexpressed human umbilical vein ECs suggested TFEB inhibits NF-κB (nuclear factor-kappa B) signaling. Indeed, luciferase assay shows TFEB suppresses NF-κB transcriptional activity. Mechanistically, TFEB suppresses IKK (IκB kinase) activity to protect IκB-α from degradation, leading to reduced p65 nuclear translocation. Inhibition of IKK by PS-1145 abolished TFEB silencing-induced inflammation in human umbilical vein ECs. Lastly, we identified KLF2 (Krüppel-like factor 2) upregulates TFEB expression and promoter activity. Laminar flow experiment showed that KLF2 is required for TFEB induction by laminar flow and TFEB is an anti-inflammatory effector downstream of laminar flow-KLF2 signaling in ECs.

Conclusions—

These findings suggest that TFEB exerts anti-inflammatory effects in diabetic mice and such function in ECs is achieved by inhibiting IKK activity and increasing IκBα level to suppress NF-κB activity. KLF2 mediates TFEB upregulation in response to laminar flow.
Rivaroxaban in Acute Coronary Syndromes: We Have a Compass and an Atlas, But Where Are We Headed?
Authors: Harrison Robert W. , Newby L. Kristin
Published: February 2019

See Article by
Gibson et al
Abstract 104: AAV-Exosomes: A Novel Platform for Myocardial Gene Delivery for Cardioprotection
Authors: Adamiak Marta , Liang Yaxuan , Mathiyalagan Prabhu , Agarwal Neha , Kohlbrenner Erik , Jha Divya , Chepurko Elena , Jeong Dongtak , Ceholski Delaine , Dubois Nicole , Hajjar Roger , Sahoo Susmita
Published: February 2019

Introduction:
Adeno-associated viruses (AAVs) are viral vectors of choice for delivering genes for long-term expression due to their safety in clinics. However, pre-existing immunity to AAVs from naturally present neutralizing antibodies (NAbs, present in between 60% and 90% of population) poses a significant challenge for AAV-mediated gene delivery. NAbs prevent AAVs from infecting target cells, greatly reducing transduction efficiency and, therefore, the clinical efficacy. Thus, it is essential to develop novel AAV-based vectors that circumvent the effect of NAbs.

Objectives:
We aimed to investigate the ability of exosome-encapsulated AAVs (AAVExo) to evade NAbs and serve as a highly efficient gene transfer tool for cardiovascular therapeutics.

Methods and Results:
We developed a multi-step purification strategy using iodixanol density gradient to isolate AAVExo with minimal contamination from free AAVs (AAV1, 6 or 9). Biochemical assays, flow cytometry, IVIS Spectrum
in vivo
optical imaging and echocardiography were used to detect AAV-mediated gene delivery and evaluate cardiac function. AAV6Exo-mCherry and AAV9Exo-FLuc were resistant to NAbs and significantly improved expression of mCherry and firefly luciferase (FLuc) both in mouse and human (iPSC-derived) cardiomyocytes
in vitro
, and in murine hearts
in vivo
(in nude mice preinjected with NAbs), compared to free AAVs. To test the therapeutic efficacy of AAVExo-mediated gene delivery in the presence of NAbs, we injected AAV9-SERCA2a or AAV9Exo-SERCA2a into post-MI hearts of nude mice preinjected with IVIg (human intravenous immunoglobulin) preparation. Hearts treated with AAV9Exo-SERCA2a had significantly improved cardiac function compared to those treated with free AAV9-SERCA2a (%EF 62.6 ± 9.8 vs. 28.4 ± 5.3, respectively; 2 weeks after surgery).

Conclusion:
Delivery of AAVs protected by carrier exosomes is a promising approach to circumvent the issue of NAbs in AAV-based gene therapy, which can be used in the entire population of patients and may result in higher gene delivery efficacy.
Abstract 117: Regulation of Cardiac Mitochondrial Function by Chaperone Mediated Autophagy
Authors: Chen Yun , Lee Jaehoon , Zheng Min , Paulino Victor , Garcia Christian , Matute Julio M , Li Hong , Kaludercic Nina , Owusu-Ansah Edward , Cuervo Ana Maria , Kitsis Richard N
Published: February 2019

Macroautophagy is required for normal cardiac function. Chaperone mediated autophagy (CMA) is distinct in that substrates are transported to lysosomes in complex with Hsc70, rather than within autophagosomes; restricted to cytoplasmic proteins; and selectively marked with a CMA-targeting motif. The role of CMA in the heart has not been assessed.

We determined that CMA is induced in the heart by pressure overload with kinetics that differ markedly from those of macroautophagy in response to the same stimulus. In CMA, cargo is imported into the lysosome by LAMP2A, which is necessary, specific, and rate-limiting for this process. Accordingly, we created knockout mice lacking LAMP2A in cardiomyocytes to inactivate CMA specifically in those cells. Unexpectedly, these mice exhibited resistance to stress-induced cardiac dysfunction in both pressure overload and myocardial infarction models. Functional assessment of cardiac mitochondria from L2AKO mice and MEFs exhibited increased rates of O
2
consumption and ATP generation. Mitochondrial proteomics showed increased levels of ATP5L, a component of mitochondrial Complex V (ATP synthase). Overexpression of ATP5L phenocopied LAMP2A KO with respect to augmentation of mitochondrial function. Conversely, knockdown of ATP5L reversed increases in mitochondrial function resulting from L2AKO. We showed that ATP5L, which possesses a KFERQ motif, is a
bona fide
CMA substrate as it interacts with Hsc70 in a KFERQ-dependent manner; and localizes to lyososmes in a LAMP2A-dependent manner when CMA is induced. ATP5L is thought to mediate oligomerization of Complex V to enhance ATP production. In fact, we observed increases in complex V oligomerization in L2A KO mouse hearts in both basal and TAC-induced states.

In summary, cardiomyocyte-specific inhibition of CMA, which renders mice resistant to stress-induced cardiac dysfunction, augments cardiac mitochondrial function through accumulation of ATP5L. Conversely, activation of CMA by cardiac stress may function as a new pathway to heart failure.
Abstract 206: Electrical Stimulation Enhances the Therapeutic Potential of Pediatric Cardiac Progenitor Cells
Authors: Maxwell Joshua , Zaladonis Carly , Jacobs Daniel , Davis Michael
Published: February 2019

Nearly 1 in every 120 children born has a congenital heart defect, and many of these children go on to develop heart failure (HF). The emergence of cardiovascular regenerative medicine as a potential therapeutic strategy for pediatric HF has provided new avenues for treatment. Stem cells such as cardiac progenitor cells (CPCs) have shown promise in restoring cardiac function in adult pathologies; however, few studies have focused on the pediatric population. Recently, pediatric CPCs >1year old have been shown to have reduced efficacy compared to younger cells. This indicates a need to enhance the regenerative potential of this population of cells for an autologous treatment for HF in pediatric patients. The regenerative potential of adult stem cells can be enhanced by
ex vivo
manipulation, and electrical stimulation (ES) is one such treatment we have examined in pediatric CPCs. We have previously shown that pediatric CPCs (1-5 years old) respond to ES by initiating intracellular Ca
2+
oscillations, and this induces phenotypic and genotypic changes in these cells. We then sought to examine the regenerative capacity of pediatric CPCs after conditioning with ES. In this study, we characterized the ES-induced changes in pediatric CPCs and showed they had a positive effect on the regenerative potential of these cells. ES-treated CPCs resulted in improved RV function following delivery compared with untreated CPCs in a juvenile rat model of right ventricular HF. Analysis of Ca
2+
handling in CPCs after chronic ES showed that these cells do not develop Ca
2+
transients or contractility similar to adult myocytes, and likely do not directly contribute to the contractility of the heart after injection. To determine if the effects were due to paracrine signaling, we tested the ability of conditioned media from these cells to regulate the function of failing cardiac myocytes. Isolated failing myocytes treated with conditioned media from CPCs showed improved Ca
2+
handling and contractility compared to untreated controls. Our data suggest that ES represents a novel treatment to enhance the therapeutic potential of pediatric CPCs, and the means of action of the modified cells is in part through a paracrine effect on resident cardiomyocytes.
Abstract 207: In Vitro Modeling of Variable Heart Diseases Due to Lamin A/C Mutation via Patient induced Pluripotent Stem Cell Derived Cardiomyocytes
Authors: Mehrabi Mehrsa , Asad Mira , Becker Danielle , Widyastuti Halida , Nguyen Cecilia , Mccarthy Linda A , Zaragoza Michael V , Grosberg Anna
Published: February 2019

According to numerous fundamental studies, a mutated Lamin A/C gene (
LMNA
) can cause dilated cardiomyopathy, arrhythmia, and heart failure, yet the practical diagnosis and treatment for them is scarce. The
LMNA
encodes Lamin A/C proteins that make mesh like nuclear lamina under the nuclear envelope, and they interact to impact the nuclear shape, stability, protein transduction, and DNA replication. Although, almost all nucleated cells in the human body have Lamin A/C in the nuclear lamina, we study patients who have
LMNA
mutation with the main pathologies in the heart. Therefore, our goal is to understand the mechanisms of how an
LMNA
mutation can cause exclusively heart diseases. We hypothesize that an
in vitro
model using induced Pluripotent Stem Cell (iPSC)-derived cardiomyocytes can differentiate between a population of patients whose major phenotype presentation is heart disease due to an
LMNA
mutation from the control population in structural and functional mechanisms. We have used
in vitro
tissue engineering techniques to interrogate the iPSC-derived cardiomyocytes tissue structures and function. PATIENT’s and CONTROL’s skin fibroblasts were reprogrammed to iPSCs and differentiated to cardiomyocytes. The PATIENT and CONTROL iPSC-derived cardiomyocytes were immunostained, imaged and quantified. Moreover, for the functional assay, iPSC- derived cardiomyocytes were evaluated by the “Heart-on-a-Chip” device based on the muscular thin film technology. The dynamics of the PATIENTs and CONTROLs tissue constructs were recorded and analyzed for frequency and stress as a function of time. Our results demonstrate that it is possible to construct an iPSC-derived cardiomyocyte
in vitro
model that differentiates between our CONTROL and PATIENT population and shows significant differences between iPSC-derived cardiomyocytes of PATIENTs and CONTROLs in frequency and active stress. By further investigating the structural and functional differences between PATIENTs and CONTROLs iPSC-derived cardiomyocytes, we can find the downstream mechanisms of LMNA mutation consequences in heart and find the proper treatments through drug screening and the other techniques by monitoring the function and structure of the iPSC-derived cardiomyocytes.
Abstract 208: miR-34a Regulates the Secretome Production and Functional Potential of Human Cardiac Progenitor Cells by Heat Shock Factor
Authors: Mishra Rachana , Sharma Sudhish , Cheng Wen C , Saha Progyaparamita , Kingsbury Tami J , Civin Curt I , Kaushal Sunjay
Published: February 2019

Background:
We have identified that chronological aging affects the myocardial function and attenuation of LV remodeling potential of Human cardiac progenitor cells (CPCs). miR-34a expression is increased in adult hearts and promotes age-associated cell death. We studied the effect of miR-34a on the cellular properties and functional potential of cardiac progenitor cells (CPCs).

Hypothesis:
We hypothesize that modulation of miR-34a in CPCs can rejuvenate them and enhance their functional potential by controlling their secretome and exosome cargo through heat shock pathway.

Methods:
Human specimens were obtained during routine cardiac surgical procedures from right atrial appendage tissue discarded from neonates (<1 month) and adults (>40 years) patients and c-kit
+
Cardiac progenitor cells (CPCs) were generated.

Results:
Here we are reporting a novel finding that knock down (KD) of miR-34a in adult-derived CPCs (aCPCs) significantly increases their cell survival, cytokine secretion by directly binding to the PDGFB, HGF1, SCF, IGF1 and FGF and affect the exosome number and their cargo via heat shock factor. The transplantation of miR34a-KD-aCPCs in myocardial infarcted (MI) rodent model significantly improved left ventricle function as analyzed by echocardiography and reduced fibrosis in comparison to the transplantation of aCPCs. Previously, we have showed that heat shock factor 1 (HSF1) is the key chaperone pathway that affects CPC’s growth potential and functional activity. Here we are showing that up-regulation of miR-34a in CPCs are attributed towards down regulation of HSF1 and their exosome secretion.

Conclusion:
CPCs improve myocardial function by their secretome, which is directly and indirectly (HSF1) controlled by miR-34a during chronological aging

.
Abstract 213: Sarcolemma Genes Related to the Transverse Tubule Structure Are Mostly Not Expressed in Differentiating Human iPSC-derived Cardiomyocytes
Authors: Vazquez Eliseo T , Torres Barba David , Sharma Janhavi S , Cortez-Toledo Elizabeth , Zhang Xiaodong , De la Cruz Cabrera Omar , Lopez Javier E
Published: February 2019

Background:
Transverse tubules (TT) are tunnel-like extensions of sarcolemma studded with ion channels coupling excitation, through the cytoplasm, to contraction in the sarcomere of matured cardiomyocytes (CMs). Expression timing of sub-cellular TT-related genes (TT-rgs) in individual human iPSC-derived CMs (hiPSC-CMs) has not yet been reported.

Objective:
Map out the gene program of TT-rgs by sub-cellular locations during hiPSC-CM differentiation using single-cell transcriptomics (scRNA Seq.).

Methods:
hiPSC-CMs were differentiated from 2 commercially available lines using sequential GSK3 and Wnt signaling inhibition. The %CMs were assayed by flow cytometry for sarcomeric myosin heavy chain protein, a CM biomarker. Baseline cells (iPSCs, n=24) and day 14 (n=45), 30 (n=64), and 60 (n=5) post differentiation cells were sampled for scRNA Seq. using the Fluidigm C1 platform. We categorized 73 TT-rgs by cellular location of the coded protein (i.e. sarcomere, cytoplasm or sarcolemma). The expression pattern for each location was categorized as induced, repressed, or neither based on the median transcript per million for each individual cell normalized to iPSCs.

Results:
CMs increased from <1% in iPSCs to 21%, 37% and 59% at D14, D30 and D60 post differentiation (adj. p <0.03 by ANOVA), respectively. 33 genes (45%) had no detectable RNA while 21 (29%) were induced to D60. See table.

Conclusion:
Single-cell transcriptomes in differentiating hiPSC-CMs revealed a discordance between sarcolemma (mostly not expressed) and sarcomeric genes being induced. Defining timing and within cell variability of TT sub-cellular genes will be critical to understand human CM maturation.
Abstract 220: Adipocyte-Derived Exosome-Containing miRNAs Exacerbated Cardiac Ischemia/Reperfusion Injury in Diabetic Mice
Authors: Gan Lu , Xie Dina , Zhao Dajun , Gao Erhe , Koch Walter J. , Wang Yajing , Ma Xinliang
Published: February 2019

Objective:
Diabetes significantly exacerbates myocardial ischemia/reperfusion (MI/R) injury. However, the molecules mediate the crosstalk between adipose tissue and heart is unclear. The current study investigated whether exosomes released by adipose tissue, the primary source for circulating exosomes, are responsible for aggravating myocardial ischemia/reperfusion injury in diabetic mice.

Methods and results:
6-8 week-old male C57B6/J mice were fed with normal diet (ND) or high fat diet (HFD) for 12 weeks to induce diabetes (characterized by abnormal glucose tolerance and insulin resistance). Exosomes were purified from ND or HFD murine serum and from culture medium of ND or HFD primary adipocytes. Equal amount of exosomes were intramyocardially injected to non-diabetic mice prior to MI/R. Myocardial functional assay showed that the exosomes purified from ND serum and culture medium of ND primary adipocytes attenuates MI/R injury (p<0.05 vs. MI/R). In contrary, the exosomes from HFD serum and culture medium of HFD primary adipocytes markedly depressed cardiac function, enlarged myocardial infarction size and increased cardiomyocyte apoptosis (p<0.05 vs. MI/R). To identify the specific molecules in exosomes that mediate cardiac injury, we performed Venn diagraph assay via combining the two databases (the adipocyte relevant miRNAs; serum exosomal miRNAs sequencing database in diabetic and nondiabetic mice). Candidate miRNAs were selected and validated. RT-qPCR assay confirmed that miR-17-3p, miR-127b-3p, miR-130b-3p, miR-345-5p and miR-532-5p were significantly elevated in exosomes isolated from HFD serum and culture medium of HFD primary adipocytes. Most importantly, these miRNAs were markedly elevated in heart tissue when exosomes isolated from HFD mice serum and culture medium of HFD primary adipocyte were injected.

Conclusion:
Our finding suggests that adipose-derived and exosome-carrying miRNAs are the likely molecules mediating the adipocyte-cardiomyocyte communication, thereby aggravating myocardial ischemia/reperfusion injury in diabetic mice.