bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrated Analysis of MicroRNA (miRNA) and mRNA Profiles Reveals Reduced Correlation between MicroRNA and Target Gene in Cancer
Authors: Li Xingsong , Yu Xiaokang , He Yuting , Meng Yuhuan , Liang Jinsheng , Huang Lizhen , Du Hongli , Wang Xueping , Liu Wanli
Published: December 2018

Background. Accumulating evidences demonstrated that microRNA-target gene pairs were closely related to tumorigenesis and development. However, the correlation between miRNA and target gene was insufficiently understood, especially its changes between tumor and normal tissues.Objectives. The aim of this study was to evaluate the changes of correlation of miRNAs-target pairs between normal and tumor.Materials and Methods. 5680 mRNA and 5740 miRNA expression profiles of 11 major human cancers were downloaded from the Cancer Genome Atlas (TCGA). The 11 cancer types were bladder urothelial carcinoma, breast invasive carcinoma, head and neck squamous cell carcinoma, kidney chromophobe, kidney renal clear cell carcinoma, kidney renal papillary cell carcinoma, liver hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, stomach adenocarcinoma, and thyroid carcinoma. For each cancer type, we firstly obtained differentially expressed miRNAs (DEMs) and genes (DEGs) in tumor and then acquired critical miRNA-target gene pairs by combining DEMs, DEGs and two experimentally validated miRNA-target interaction databases, miRTarBase and miRecords. We collected samples with both miRNA and mRNA expression values and performed a correlation analysis by Pearson method for miRNA-target pairs in normal and tumor, respectively.Results. We totally got 4743 critical miRNA-target pairs across 11 cancer types, and 4572 of them showed weaker correlation in tumor than in normal. The average correlation coefficients of miRNA-target pairs were different greatly between normal (-0.38 ~ -0.61) and tumor (-0.04 ~ -0.26) for 11 cancer type. The pan-cancer network, which consisted of 108 edges connecting 35 miRNAs and 89 target genes, showed the interactions of pairs appeared in multicancers.Conclusions. This comprehensive analysis revealed that correlation between miRNAs and target genes was greatly reduced in tumor and these critical pairs we got were involved in cellular adhesion, proliferation, and migration. Our research could provide opportunities for investigating cancer molecular regulatory mechanism and seeking therapeutic targets.
An Efficient Color Space for Deep-Learning Based Traffic Light Recognition
Authors: Kim Hyun-Koo , Park Ju H. , Jung Ho-Youl
Published: December 2018

Traffic light recognition is an essential task for an advanced driving assistance system (ADAS) as well as for autonomous vehicles. Recently, deep-learning has become increasingly popular in vision-based object recognition owing to its high performance of classification. In this study, we investigate how to design a deep-learning based high-performance traffic light detection system. Two main components of the recognition system are investigated: the color space of the input video and the network model of deep learning. We apply six color spaces (RGB, normalized RGB, Ruta’s RYG, YCbCr, HSV, and CIE Lab) and three types of network models (based on the Faster R-CNN and R-FCN models). All combinations of color spaces and network models are implemented and tested on a traffic light dataset with 1280×720 resolution. Our simulations show that the best performance is achieved with the combination of RGB color space and Faster R-CNN model. These results can provide a comprehensive guideline for designing a traffic light detection system.
Autophagy and Its Role in Protein Secretion: Implications for Cancer Therapy
Authors: Cotzomi-Ortega Israel , Aguilar-Alonso Patricia , Reyes-Leyva Julio , Maycotte Paola
Published: December 2018

Autophagy is a protein and organelle degradation pathway important for the maintenance of cytoplasmic homeostasis and for providing nutrients for survival in response to stress conditions. Recently, autophagy has been shown to be important for the secretion of diverse proteins involved in inflammation, intercellular signaling, and cancer progression. The role of autophagy in cancer depends on the stage of tumorigenesis, serving a tumor-suppressor role before transformation and a tumor-survival function once a tumor is established. We review recent evidence demonstrating the complexity of autophagy regulation during cancer, considering the interaction of autophagy with protein secretion pathways. Autophagy manipulation during cancer treatment is likely to affect protein secretion andinter-cellular signaling either to the neighboring cancer cells or to the antitumoral immune response. This will be an important consideration during cancer therapy since several clinical trials are trying to manipulate autophagy in combination with chemotherapy for the treatment of diverse types of cancers.
Empirical Path‐Loss Modeling and a RF Detection Scheme for Various Drones
Authors: Jeong Won Ho , Choi Hong-Rak , Kim Kyung-Seok
Published: December 2018

This paper presents a path‐loss model based on a radio‐frequency (RF) detection scheme for various drones using 5G aerial communication over an industrial, scientific, and medical radio band (ISM band) network. We considered three communication modes of the ISM band for the channel characteristics analysis: the DJI Enhanced Spread Spectrum Technology (DESST) protocol, Wi‐Fi, and Bluetooth. The drone signal detection scheme extracts the drone signal from the environment mixed with the general signal. The drone DESST signal is identified through cross‐correlation of the received signal. The Wi‐Fi and Bluetooth signals are identified with the singular‐value decomposition (SVD) algorithm by using the hopping characteristics. General and drone Wi‐Fi signals are separated by in‐phase/quadrature (I/Q) phase analysis over the measurement time. The windowed received signal strength indicator (RSSI) moving detection (WRMD) analysis identifies the drone Bluetooth signal according to the movement of the drone. The detected drone signal is channel modeled by the horizontal distance d according to the altitude θ. Finally, they verify their model by a ray‐tracing simulation similar to the real environment. The model provides a simple and accurate prediction for designing future aerial communications systems according to changes in drone movement.
The Role of Zinc and Zinc Homeostasis in Macrophage Function
Authors: Gao Hong , Dai Wei , Zhao Lu , Min Junxia , Wang Fudi
Published: December 2018

Zinc has long been recognized as an essential trace element, playing roles in the growth and development of all living organisms. In recent decades, zinc homeostasis was also found to be important for the innate immune system, especially for maintaining the function of macrophages. It is now generally accepted that dysregulated zinc homeostasis in macrophages causes impaired phagocytosis and an abnormal inflammatory response. However, many questions remain with respect to the mechanisms that underlie these processes, particularly at the cellular and molecular levels. Here, we review our current understanding of the roles that zinc and zinc transporters play in regulating macrophage function.
Numerical Modeling of the Conductivity of the Particle Monolayer with Reduced Size
Authors: Liu Yuxuan , Wang Jiandong , Guo Jianchun , Zhu Haiyan , Zeng Jie
Published: December 2018

Fractures filled with a proppant monolayer play an important role in the hydraulic fracture network. Predicting the conductivity of these fractures is the basis of fracture network optimization. However, little attention has been paid to the conductivity of the proppant monolayer. The change of conductivity under various conditions is currently not fully understood. Therefore, in this paper, the conductivity variation under different conditions are simulated. The reduction of particle size was calculated by existing analytical models. The permeability variation was calculated through computational fluid dynamics (CFD) combined with COMSOL Multiphysics. The controlling factors of conductivity under a proppant monolayer were identified. Simulation results indicate that elastic parameters, closure pressure, and proppant distribution have significant influence on conductivity, while creep parameters, such as rock viscosity and time, have limited influence on conductivity. Moreover, the changes in permeability, porosity, and tortuosity with variation of embedment were analyzed. Results indicated that with an increase in embedment, the permeability and porosity decrease as expected. The main reduction (nearly half) emerges in the first 20% of proppant embedment. Furthermore, the permeability of a single particle deviates largely from the prediction of Carman-Kozeny (CK) equation. The tortuosity of proppant particle increases with a decrease in particle size due to embedment. A modification of the Carman-Kozeny equation is proposed to address this influence.
Automatically Traceback RDP‐Based Targeted Ransomware Attacks
Authors: Wang ZiHan , Liu ChaoGe , Qiu Jing , Tian ZhiHong , Cui Xiang , Su Shen
Published: December 2018

While various ransomware defense systems have been proposed to deal with traditional randomly‐spread ransomware attacks (based on their unique high‐noisy behaviors at hosts and on networks), none of them considered ransomware attacks precisely aiming at specific hosts, e.g., using the common Remote Desktop Protocol (RDP). To address this problem, we propose a systematic method to fight such specifically targeted ransomware by trapping attackers via a network deception environment and then using traceback techniques to identify attack sources. In particular, we developed various monitors in the proposed deception environment to gather traceable clues about attackers, and we further design an analysis system that automatically extracts and analyze the collected clues. Our evaluations show that the proposed method can trap the adversary in the deception environment and significantly improve the efficiency of clue analysis. Furthermore, it also helps us trace back RDP‐based ransomware attackers and ransomware makers in the practical applications.
Preparation of a Self‐Lubricating Cu/h‐BN Coating on Cemented Carbide
Authors: Cao Tongkun , Zhu Zhibin , Liu Yajun
Published: December 2018

In this work, a Cu/h‐BN self‐lubricating coating was prepared on cemented carbide by electrospark deposition (ESD). The microstructure and properties of the coating were examined. The results showed that no decomposition of or reactions with h‐BN occurred. As the h‐BN content and capacitance increased, the number of pores and microcracks in the coatings increased. Additionally, as the capacitance increased, the electrode mass loss increased. However, the specimen mass increased first and then decreased. The coating thickness was affected by the capacitance, deposition time, and volume ratio of h‐BN to Cu. The results exhibited were consistent over the tests. The self‐lubricating coating exhibited excellent tribological behavior under the test conditions, and the worn surface showed features consistent with shear slippage and abrasive wear.
Tensor Decomposition for Multiple‐Instance Classification of High‐Order Medical Data
Authors: Papastergiou Thomas , Zacharaki Evangelia I. , Megalooikonomou Vasileios
Published: December 2018

Multidimensional data that occur in a variety of applications in clinical diagnostics and health care can naturally be represented by multidimensional arrays (i.e., tensors). Tensor decompositions offer valuable and powerful tools for latent concept discovery that can handle effectively missing values and noise. We propose a seamless, application‐independent feature extraction and multiple‐instance (MI) classification method, which represents the raw multidimensional, possibly incomplete, data by means of learning a high‐order dictionary. The effectiveness of the proposed method is demonstrated in two application scenarios: (i) prediction of frailty in older people using multisensor recordings and (ii) breast cancer classification based on histopathology images. The proposed method outperforms or is comparable to the state‐of‐the‐art multiple‐instance learning classifiers highlighting its potential for computer‐assisted diagnosis and health care support.
Usual Interstitial Pneumonia Pattern in the Lower Lung Lobes as a Prognostic Factor in Idiopathic Pleuroparenchymal Fibroelastosis
Authors: Kato Motoyasu , Sasaki Shinichi , Kurokawa Kana , Nakamura Takahiro , Yamada Tomoko , Sasano Hitoshi , Arano Naoko , Komura Moegi , Ihara Hiroaki , Nagashima Osamu , Shiota Satomi , Takahashi Fumiyuki , Takahashi Kazuhisa
Published: December 2018

Background: Idiopathic pleuroparenchymal fibroelastosis (IPPFE) is a rare interstitial pneumonia that is characterized by stiffness in both the upper lobes and pleura, which is evident on high resolution computed tomography (HRCT) of the chest. However, prognostic factors for IPPFE have not been identified yet. Objective: We aimed to investigate the clinical prognostic factors affecting survival in patients with IPPFE. Methods: Between April 2009 and September 2017, we enrolled 36 patients who were clinically diagnosed with IPPFE, using HRCT. These patients were classified as either short survival (dead within 12 months from the diagnosis of IPPFE) or long survival (survived for greater than 12 months) groups. We retrospectively analyzed the clinical characteristics, serum markers, pulmonary function test results, and HRCT findings. Results: Twelve patients were classified into the short survival and 24 were categorized into long survival categories. At the time of diagnosis, the incidence of coexistence of a usual interstitial pneumonia (UIP) pattern in the lower lobes on HRCT in the short survival was significantly higher than that in the long survival. Multivariate analysis revealed that a UIP pattern in the lower lobes on HRCT was the only independent variable for poor prognosis. The median survival time from diagnosis in patients with IPPFE was 24 months. Of these patients with IPPFE, the survival time with a UIP pattern was significantly shorter than in those without a UIP pattern. Conclusion: Our findings suggest that a UIP pattern in the lower lobes at the time of diagnosis was an independent prognostic factor for IPPFE.
Difficult Patient Behavior in Dialysis Facilities
Authors: Janosevic Danielle , Wang Aileen X. , Wish Jay B.
Published: December 2018

Difficult behavior exhibited by dialysis patients is a spectrum that includes nonadherence, verbal or physical abuse, and threatening acts. Such behaviors may lead to harmful consequences to the patient, other patients, the facility, and staff and can culminate in involuntary discharge. It is important to recognize that these “difficult behaviors” may be due to underlying psychosocial or medical issues, which places an onus on care providers to explore further. According to the Conditions for Coverage (CfC) for dialysis facilities, it falls upon the medical director to coordinate and oversee policies for patient satisfaction, patient safety and rights, involuntary discharges, and adverse events and outcomes. Thus, medical directors are liable for their own actions, and their staff for which they have oversight, for harm or perceived harm to patients in response to difficult behaviors. Guidelines to deal with specific patient behavior scenarios have been published by the Decreasing Dialysis Patient Conflict National Task Force of the Forum of end-stage renal disease (ESRD) Networks. The common denominator for these difficult scenarios is impaired communication, and the majority of patient concerns involve issues with staff, policies, treatments, and diet. Involuntary discharge of a patient should always be viewed as a last resort, and there is a structured process described in the CfC that requires the involvement of the respective ESRD Network and the facility medical director. As physicians, we are bound by ethical and growing legal obligations to act in an appropriate, ethical, and fair manner to patients who are considered to be “difficult.”
Can the Mixed Virtual Reality Simulator Into D’mentia Enhance Empathy and Understanding and Decrease Burden in Informal Dementia Caregivers?
Authors: Jütten Linda Helena , Mark Ruth Elaine , Sitskoorn Margriet Maria
Published: December 2018

Objective: To evaluate whether the mixed virtual reality dementia simulator training Into D’mentia increased informal caregivers’ understanding for people with dementia, their empathy, sense of competence, relationship quality with the care receiver, and/or decreased burden, depression, and anxiety. Methods: A quasi-experimental longitudinal study with an intervention group (n = 145) and a control group (n = 56) was conducted. All participants were informal caregivers of people with dementia. They completed six questionnaires and semi-structured interviews 1 week before as well as 1 week, 2.5 months, and 15 months after the training. Data were analyzed on both group and individual level using linear mixed model analyses and Reliable Change Indices. Results: Eighty-five percent of the participants in the intervention group found the intervention useful; 76% said they had changed their approach to caregiving, and 61% stressed that the intervention had increased their understanding of dementia. No significant differences were found between the two groups over time regarding empathy, sense of competence, relationship quality with the care receiver, burden, depression, and anxiety, at either group or individual level. Conclusion: Caregivers indicated that the Into D’mentia intervention improved their understanding of dementia, that they had learned to be more patient, to take things more slowly, and to focus on positive aspects of caregiving. However, no significant change was found on the variables assessed via the questionnaires. Future research can consider enriching this intervention with other aspects such as more educational material, more simulations, and group sessions, tailored to the individual caregiver and his/her situation, and examine whether these new interventions yield change on questionnaires. These new, more personalized interventions for dementia caregivers could help caregivers to better understand the persons with dementia they care for and to ultimately enhance the well-being of both caregivers and persons with dementia.
Neurofilament Light in Serum and Cerebrospinal Fluid of Hip Fracture Patients with Delirium
Authors: Halaas Nathalie Bodd , Blennow Kaj , Idland Ane-Victoria , Wyller Torgeir Bruun , Ræder Johan , Frihagen Frede , Staff Anne Cathrine , Zetterberg Henrik , Watne Leiv Otto
Published: December 2018

Background: Delirium is associated with new-onset dementia, suggesting that delirium pathophysiology involves neuronal injury. Neurofilament light (NFL) is a sensitive biomarker for neuroaxonal injury. Methods: NFL was measured in cerebrospinal fluid (CSF) (n = 130), preoperative serum (n = 192), and postoperative serum (n = 280) in hip fracture patients, and in CSF (n = 123) and preoperative serum (n = 134) in cognitively normal older adults undergoing elective surgery. Delirium was diagnosed with the Confusion Assessment Method. Results: Median serum NFL (pg/mL) was elevated in delirium in hip fracture patients (94 vs. 54 pre- and 135 vs. 92 postoperatively, both p < 0.001). Median CSF NFL tended to be higher in hip fracture patients with delirium (1,804 vs. 1,636, p = 0.074). Serum and CSF NFL were positively correlated (ρ = 0.56, p < 0.001). Conclusion: Our findings support an association between neuroaxonal injury and delirium. The correlation between serum and CSF NFL supports the use of NFL as a blood biomarker in future delirium studies.
Down-Regulation of Laminin (LN)- α5 is Associated with Preeclampsia and Impairs Trophoblast Cell Viability and Invasiveness Through PI3K Signaling Pathway
Authors: Zhang Xue Mei , Xiong Xi , Tong Chao , Li Qin , Huang Shuai , Li Qing Shu , Liu Ya Ming , Li Hai Ying , Baker Phillip , Shan Nan , Qi Hong Bo
Published: December 2018

Background/Aims: Preeclampsia (PE) is a gestational disorder defined as hypertension and proteinuria, which is deemed a major cause of maternal and neonatal mortality and morbidity worldwide. The aim of this study was to investigate the expression patterns of placental laminin (LN)-α5 expression in normal and PE pregnancies, as well as evaluating the effects of LN-α5 on trophoblast proliferation, apoptosis, and invasion. Methods: LN-α5 expression levels were examined by reverse-transcriptase polymerase chain reaction (RT-PCR), and further confirmed by western blotting and immunofluorescence staining. Cell proliferation and apoptosis were measured by CCK-8 assay and flow cytometry. Cell invasion was assessed by matrigel-based transwell assay. LN-α5 DNA methylation in placentas was determined by bisulfite sequencing PCR (BSP). Results: LN-α5 expression levels in PE placentas were significantly lower than that of normal pregnancies. Deficiency in LN-α5 expression resulted in decreased trophoblast proliferation and invasion but increased cell apoptosis, meanwhile, PI3K/AKT/mTOR signaling pathway was impaired by LN-α5 silencing. LN-α5 promoter methylation didn’t show significant difference between PE and normal placentas. Conclusion: LN-α5 downregulation is associated with PE placenta and impairs trophoblast viability and invasiveness, which could be a causative factor of PE pathogenesis.
Clinicopathologic Features and Prognostic Factors in Alpha-Fetoprotein-Producing Colorectal Cancer: Analysis of 78 Cases
Authors: Feng Yang , Li Yaqi , Dai Weixing , Mo Shaobo , Li Qingguo , Cai Sanjun
Published: December 2018

Background/Aims: Alpha-fetoprotein-producing colorectal cancer (AFPP-CRC) is quite rarely seen. This study aimed to elucidate the clinicopathologic characteristics and prognostic factors of AFPP-CRC. Methods: Among 5,051 colorectal cancer patients receiving surgery in the Fudan University Shanghai Cancer Center from 2006 to 2016, we identified 78 patients with elevated serum level of AFP (> 10 µg/L) preoperatively. A propensity score matching (PSM) analysis was performed which matched 75 AFPP-CRC patients to the same number of AFP-negative colorectal cancer (AFPN-CRC) patients. Kaplan-Meier curves were compared using the log-rank test and multivariable analysis was performed to evaluate the effect of AFP-positivity while adjusting confounding factors. 27 patients were available for immunohistochemical analysis. We conducted functional experiments to characterize the tumorigenicity of AFP. Results: Patients with AFPP-CRC had a significantly higher incidence of advanced TNM stage and liver metastasis. Overall survival was significantly different between two groups before and after PSM, and AFP-positivity was one of the strongest predictors of overall survival in the multivariable model (HR 4.11, CI 95%: 1.43-11.76, p = 0.009) after PSM. We further investigated prognostic factors affecting prognosis in AFPP-CRC and found that the presence of liver metastasis was the only independent prognostic factor (HR 4.95, CI 95%: 1.48-16.48, p = 0.009). AFP expression was significantly positively correlated with HGF and c-Met expression. Transwell invasion assay revealed significantly increased cell motility with AFP overexpression. Conclusion: AFP-positivity is a significant negative predictor of overall survival in patients with colorectal cancer, which may be mediated by HGF/c-Met signaling pathway.
Gliquidone Alleviates Diabetic Nephropathy by Inhibiting Notch/Snail Signaling Pathway
Authors: Tian Hengyu , Yang Junbo , Xie Zhuochao , Liu Jialin 
Published: December 2018

Background/Aims: Diabetic nephropathy is a common complication of diabetes. This study explored the renal protective effect and possible mechanism of gliquidone in mice with diabetic nephropathy. Methods: Animal model of diabetic nephropathy was established in KKAy mice. The renal protective effect of gliquidone was studied by evaluating the kidney function through measures of urinary protein, blood urea nitrogen (BUN), serum creatinine (Scr) and serum triglyceride (TG) that were performed using an automatic biochemical analyzer. The levels of oxidative stress indicators, such as nitric oxide (NO), superoxide dismutase (SOD) and malondialdehyde (MDA), were evaluated in renal tissue homogenates using the automatic biochemical analyzer. The inhibitory effect of gliquidone on renal interstitial fibrosis and its association with Notch / Snail1 signaling pathway in diabetic nephropathy was investigated using molecular biological techniques. Results: It was found that low-, medium- and high-dose gliquidone improved the mice’s general health condition, such as mental status, fur condition, eating, and drinking. Gliquidone reduced the body weight and the kidney weight /body weight ratio of mice. Gliquidone improved the kidney function, indicated by reductions in urinary protein, blood urea nitrogen, and serum creatinine and triglyceride. Gliquidone treatment increased levels of nitric oxide and superoxide dismutase, but decreased level of malondialdehyde. The expression of Jagged1/Notch1/hes1/Snail1/α-SMA decreased, while the expression of E-cadherin increased in gliquidone-treated kidneys. High dose gliquidone showed the best effect, one that was similar to that of the positive control drug irbesartan. Conclusion: Taken together, our results suggested that gliquidone can ameliorate the diabetic symptoms of diabetic nephropathy through inhibiting Notch / Snail1 signaling pathway, improving anti -oxidative response and delaying renal interstitial fibrosis. The efficacy of gliquidone is dose-dependent.
Gastric Adenocarcinoma Predictive Long Intergenic Non-Coding RNA Promotes Tumor Occurrence and Progression in Non-Small Cell Lung Cancer via Regulation of the miR-661/eEF2K Signaling Pathway
Authors: Gu Haiting , Chen Junfeng , Song Yukang , Shao Haiyan
Published: December 2018

Background/Aims: Long non-coding RNAs (lncRNAs) play vital roles in carcinogenesis as oncogenes or tumor suppressor genes. This study explored the biological function of lncRNA gastric adenocarcinoma predictive long intergenic non-coding RNA (GAPLINC) in human non-small cell lung cancer (NSCLC). Methods: GAPLINC expression in NSCLC specimens and cell lines was detected by qRT-PCR and Western blot. The effect of GAPLINC on cell proliferation was investigated using CCK8-assay, colony formation assay, and xenograft model. The effects of GAPLINC on apoptosis and cell cycle were determined using flow cytometry. The mechanism of GAPLINC involved in NSCLC was explored using Western blot, luciferase reporter assay, and RNA fluorescence in situ hybridization. Results: We found that GAPLINC expression was up-regulated in NSCLC tissues and cell lines. Overexpression of GAPLINC was associated with poor prognosis in patients with NSCLC. Silencing of GAPLINC significantly inhibited cell proliferation, promoted apoptosis, and induced cell cycle arrest in the G0/G1 phase. Results from xenograft transplantation showed that GAPLINC silencing inhibited the tumor growth in vivo. Interestingly, GAPLINC silencing decreased the expression of eukaryotic elongation factor-2 kinase (eEF2K) protein both in vivo and in vitro. Bioinformatic analysis and luciferase reporter confirmed that miR-661 targeted GAPLINC and eEF2K 3’-UTR and was negatively correlated with the expression of GAPLINC and eEF2K. Conclusion: Our findings indicate that GAPLINC promotes NSCLC tumorigenesis by regulating miR-661/eEF2K cascade and provide new insights for the pathogenesis underlying NSCLC and potential targets for therapeutic strategy.
Relationship Between the Efficacy of Cardiac Cell Therapy and the Inhibition of Differentiation of Human iPSC-Derived Nonmyocyte Cardiac Cells Into Myofibroblast-Like Cells
Authors: Gao Ling , Yang Libang , Wang Lu , Geng Zhaohui , Wei Yuhua , Gourley Glenn , Zhang Jianyi
Published: December 2018

Rationale:

Myofibroblasts are believed to evolve from precursor cells; however, whether noncardiomyocyte cardiac cells (NMCCs; ie, endothelial cells, smooth muscle cells, pericytes, and fibroblasts) that have been derived from human-induced pluripotent stem cells (hiPSCs) can transdifferentiate into myofibroblast-like cells, and if so, whether this process reduces the efficacy of hiPSC-NMCC therapy, is unknown.

Objective:

To determine whether hiPSC-NMCCs can differentiate to myofibroblast-like cells and whether limiting the transdifferentiation of hiPSC-NMCCs can improve their effectiveness for myocardial repair.

Methods and Results:

When endothelial cells, smooth muscle cells, pericytes, and fibroblasts that had been generated from hiPSCs were cultured with TGF-β (transforming growth factor-β), the expression of myofibroblast markers increased, whereas endothelial cell, smooth muscle cell, pericyte, and fibroblast marker expression declined. TGF-β–associated myofibroblast differentiation was accompanied by increases in the signaling activity of Smad, Snail, and mTOR (mammalian target of rapamycin). However, measures of pathway activation, proliferation, apoptosis, migration, and protein expression in hiPSC-endothelial cell–derived, smooth muscle cell-derived, pericyte-derived, and fibroblast-derived myofibroblast-like cells differed. Furthermore, when hiPSC-NMCCs were transplanted into the hearts of mice after myocardial infarction, ≈21% to 35% of the transplanted hiPSC-NMCCs expressed myofibroblast markers 1 week later, compared with <7% of transplanted cells (
P
<0.01, each cell type) in animals that were treated with both hiPSC-NMCCs and the TGF-β inhibitor galunisertib. Galunisertib coadministration was also associated with significant improvements in fibrotic area, left ventricular dilatation, vascular density, and cardiac function.

Conclusions:

hiPSC-NMCCs differentiate into myofibroblast-like cells when cultured with TGF-β or when transplanted into infarcted mouse hearts, and the phenotypes of the myofibroblast-like cells can differ depending on the lineage of origin. TGF-β inhibition significantly improved the efficacy of transplanted hiPSC-NMCCs for cardiac repair, perhaps by limiting the differentiation of hiPSC-NMCCs into myofibroblast-like cells.
Abstract 008: Chronic Central Melanocortin 4 Receptor Activation Attenuates Cardiac Dysfunction After Myocardial Infarction In Rats
Authors: Gava Fabio N , da Silva Alexandre A , Hall John E , do Carmo Jussara M
Published: December 2018

The melanocortin pathway plays an important role in multiple physiological functions besides its effect on energy homeostasis. In the present study we tested whether activation of the brain melanocortin 4 receptors (MC4R) confers protection against cardiac dysfunction after myocardial infarction (MI). Male Sprague-Dawley rats at 12 weeks of age were implanted with blood pressure telemetry transmitters and an intracerebroventricular (ICV) cannula was inserted into the lateral ventricle. After 10 days of recovery, food intake, mean arterial pressure (MAP) and heart rate (HR) were measured 24-hrs/day by telemetry and cardiac function was assessed by echocardiography (VEVO 3100
@
). After stable baseline measurements for 4 days, the left coronary descending artery was permanently ligated and vehicle (n=6) or the MC4R agonist (MTII, 10 ng/hr, n=7) was infused ICV via osmotic minipump for 28 consecutive days. Chronic MC4R activation significantly decreased cumulative food intake (-121±17 g) and body weight (385±5 to 378±4 g) compared to vehicle treatment (-51±10 g and 358±9 to 427±9 g). Chronic MTII infusion did not significantly alter MAP and HR (113±1 to 115±2 mmHg and 356±3 to 349±9 bpm) whereas MAP and HR were slightly reduced in vehicle-treated rats with MI (109±1 to 105±3 mmHg and HR 357±3 to 324±5). Compared to vehicle, MTII infusion for 4 weeks attenuated cardiac dysfunction caused by MI as evidenced by normalization of global cardiac radial strain (39±4 to 39±3 vs. 37±4 to 25±3 %), cardiac output (117±9 to 117±8 vs. 105±6 to 83±9 ml/min) and improvement of ejection fraction (72±2 to 47±3 vs. 70±2 to 30±2 %). These results suggest that chronic central MC4R activation attenuates the progression of heart failure after MI in rats. (NHLBI-PO1HL51971, NIGMS- P20GM104357 and U54GM115428)
Abstract 021: Interleukin-21 Plays a Critical Role in Hypertension and Vascular Dysfunction
Authors: Dale Bethany L , Pandey Arvind , Chen Yuahn , Laroumanie Fanny , Itani Hana , Xiao Liang , Norlander Allison , Foss Jason , Barbaro Natalia , Van Beusecum Justin , Ikizler Alp , Alsouqi Aseel , Scott Holly M , Madhur Meena S
Published: December 2018

T cell derived cytokines, interleukin 17A (IL17A) and interferon gamma (IFNg), promote angiotensin II (Ang II)-induced hypertension and end-organ damage. Interleukin 21 (IL21), produced primarily by T follicular helper (Tfh) cells, has been shown to induce IL17A and IFNγ production from T effector cells. IL21 is also a potent activator of germinal center (GC) B cells and immunoglobulin (Ig) class switching.
We hypothesized that IL21 plays a fundamental role in hypertension and hypertensive end-organ damage through its effects on pro-inflammatory T cell polarization, vascular dysfunction, and enhanced Ig secretion.
CD4+ T cells from Ang II infused WT mice exhibit 1.5-fold increased IL21 mRNA expression and 2-fold increased IL-21 production compared to vehicle. We found IL21
-/-
mice exhibit a 30mmHg reduction in systolic blood pressure (SBP) in response to 4 weeks of Ang II infusion compared to age-matched wild type (WT) mice. Further, IFNg and IL17A production from CD8+ or CD4+ T cells, respectively, was abrogated in IL21
-/-
mice vs. WT mice infused with Ang II. We also found Tfh cells and GC B cells significantly increased in the aortas and mesenteric vessels of Ang II WT mice but not IL21
-/-
mice. Ang II induced an increase in percent of lymph node GC B cells and IgG/IgM ratio, consistent with Ig class switching. Ang II-induced increase in renal injury was also blunted in IL21
-/-
mice vs. WT mice. Mesenteric vessels from IL21
-/-
mice were protected from Ang II-induced endothelial dysfunction. IL21
-/-
mice also developed blunted aortic fibrosis and reduced smooth muscle cell hypertrophy after 4 weeks of Ang II vs. WT mice. IL-21 neutralization beginning 2 weeks after Ang II resulted in a 15 mmHg reduction in SBP and reversal of aortic inflammation and vascular endothelial dysfunction compared to isotype control treated animals. Lastly, CD4+ T cell production of IL21 positively correlated with SBP in human volunteers, and in fact, IL-21 production was significantly higher in hypertensive compared to normotensive subjects. Taken together, these studies provide compelling evidence that IL21 may function as a master cytokine in the pathogenesis of hypertension and suggest that therapies targeting IL21 may reduce hypertension and the associated end-organ damage.
Abstract 023: Redox-Driven T-Lymphocyte Inflammation Sensitizes Mice to Psychological Stress-Mediated Hypertension
Authors: Moshfegh Cassandra M , Collins Christopher W , Kohl Allison J , Elkhatib Safwan , Case Adam J
Published: December 2018

Post-traumatic stress disorder (PTSD) is a debilitating psychological disease that increases sympathoexcitation, norepinephrine (NE) outflow, and the risk of cardiovascular diseases like hypertension by >50%, but the mechanisms remain unclear. Hypertension is regulated in part due to immune system-driven inflammation, and our laboratory has previously reported that exposure to NE can modulate levels of T-lymphocyte pro-inflammatory cytokines by redox signaling mechanisms. Therefore, we hypothesized that psychological stress-induced sympathoexcitation leads to a redox-regulated increase in pro-inflammatory cytokine production from T-lymphocytes, which may impact the development of hypertension. Utilizing a mouse-model of PTSD-like psychological stress (
i.e.
social defeat), increased sympathoexcitation was confirmed by a 2.5 fold increase in splenic NE content and 4.1 fold increase in tyrosine hydroxylase (TH; p<0.05). Splenic and peripheral T-lymphocytes demonstrated increased reactive oxygen species (ROS) levels as evidenced by enhanced dihydroethidium and MitoSOX Red oxidation (p<0.05). Increases in ROS were correlated with elevated pro-inflammatory cytokines IL-17A, IL-6, and IL-2 in circulation as well as specifically within T-lymphocytes (p<0.05). Examination of hemodynamics elucidated a progressive increase in mean arterial pressure (15 mmHg, p<0.05) over the stress-induction period, correlating with the elevation of T-lymphocyte driven inflammation. This hypertension was not associated with an increase in circulating renin levels. To investigate the causal contribution of T-lymphocytes in driving this hypertension, recombination activating gene 2 (Rag2) knock-out animals, which are devoid of mature lymphocytes, were utilized. Rag2
-/-
mice showed no changes in pro-inflammatory cytokines, a 75% decrease in splenic TH levels (p<0.05), and decreased blood pressure after stress, suggesting T-lymphocytes may regulate the stress-induced hypertension. Overall, our data suggest the potential for a new paradigm involving redox control of T-lymphocytes in the regulation of psychological stress-driven hypertension.
Abstract 034: Resilience of Isolated, Perfused Cerebral Penetrating Microarterioles to Angiotensin II (Ang II) Contractions Depends on Local Generation of Prostaglandin D
2
(PGD
2
)
Authors: Li Lingli , Lai EnYin , Luo Zaiming , Davisson Robin , Welch William , Wilcox Christopher S
Published: December 2018

The brain depends on a continuous supply of blood for its oxygenation, whereas the kidney is over-perfused for its metabolic needs in order to provide sufficient plasma to form a glomerular filtrate. Thus, the brain requires resilience to vasoconstriction to prevent ischemia and vascular cognitive impairment, but the mechanisms are unclear.

Methods and Results:
Single penetrating cerebral microarterioles (CMAs, 12-18μM) were dissected from the frontal cortex and single renal afferent arterioles (RAAs, 8-12μM) from the kidney cortex to investigate the hypothesis that CMAs deploy unique mechanisms to provide resilience to Ang II vasoconstriction. Individual arteriolar genes were assessed by RNAseq or RT-PCR of endothelial cells (ECs). The mRNA for lipocalin type PGD
2
synthase (LPGDS) and the PGD 1 receptor (PD1R) were > 3,000-fold higher in CMAs than RAAs whereas RAAs expressed 3-fold more mRNA for thromboxane A
2
synthase. Both microarterioles had similar expression of AT1Rs. Endothelial cells cultured from these vessels had similar patterns of gene expression. Single isolated perfused RAAs contracted strongly with Ang II (at 10
-6
mol·l
-1
; -47 ± 2%; P<0.005) whereas CMAs were totally resistant to Ang II (0.1 ± 0.1%; NS), yet both contracted similarly to endothelin I or perfusion pressure (n = 6 per group). However, CMAs from COX 1 -/- (vs +/+) mice did contract with Ang II (-15 ± 2 vs 0.1 ± 0.1%; P<0.01) and contracted with Ang II after incubation with parecoxib (vs vehicle) to block COX2 (-7 ± 3 vs 0.1 ± 0.1%; P<0.01) or after dual blockade of COX1 + 2 (-20 ± 2%; P<0.01) or after incubation with AT-56 (vs vehicle) to block LPGDS (-20 ± 3 vs 0.1 ± 0.1%; P<0.01). During LPGDS blockade, incubation of CMAs with BW245c (stable PD1R agonist) reduced Ang II contraction > 65% (-8 ± 2%; P<0.01). In contrast, COX blockade reduced Ang II contractions of RAAs, indicating opposing effects of PGs on cerebral and renal vessels. Measurements of cerebral and renal blood flow and MAP in anesthetized mice confirmed selective renal vasoconstriction with Ang II, yet selective cerebral vasodilation with BW245c.

Perspective:
Resilience against Ang II vasoconstriction in cerebral arterioles depends on the generation of PGD
2
and could be a therapeutic target for vascular dementia and stroke.
Abstract 035: Increasing Renal AcSDKP by Eliminating the ACE N-Domain Blocks Renal Inflammation and Sodium Retention During Diabetic Nephropathy
Authors: Giani Jorge F , Eriguchi Masahiro , Bernstein Ellen A , Veiras Luciana C , Khan Zakir , Cao Duo Y , Fuchs Sebastien , McDonough Alicia A , Toblli Jorge E , Gonzalez-Villalobos Romer A , Bernstein Kenneth E
Published: December 2018

Angiotensin-converting enzyme (ACE) plays a key role in renal inflammation and sodium retention associated with diabetic nephropathy. Although most effects of ACE have been classically related to angiotensin (Ang) II synthesis, studies highlight an Ang II-independent role of ACE in inflammation. Indeed, ACE has two catalytic domains, the N- and C-domains, that can process a wide diversity of substrates besides Ang I. Here, we study the relative contributions of ACE domains to renal inflammation and sodium retention during diabetic nephropathy. Diabetes was induced with streptozotocin in wild-type (WT) mice and mice lacking either a functional ACE N-domain (NKO) or C-domain (CKO) (n=5-8). After 6 months of diabetes, we evaluated the natriuretic response to volume expansion. For this, mice were injected with 0.9% NaCl equivalent to 10% of their body weight. After 5 hours, diabetic NKO mice excreted 30% more urinary sodium in response to the saline challenge compared to diabetic WT or CKO mice (
P
<0.05). This enhanced natriuretic response was associated with a 47% reduction (
P
<0.05) of renal epithelial sodium channel (ENaC) cleaved (active) α and γ subunits. Further, diabetic NKO displayed lower levels of renal fibrosis (46% reduction,
P
<0.05), IL-1β (56% reduction,
P
<0.05), TNFα (50% reduction,
P
<0.05) and albuminuria (53% reduction,
P
<0.01) compared to WT and CKO diabetic mice. This protective phenotype was not associated with changes in renal Ang II levels (WT: 237 ± 73; NKO: 283 ± 53 CKO: 245 ± 39 fmol/g kidney,
P
=NS). We next evaluated whether the anti-inflammatory tetrapeptide N-acetyl-seryl-asparyl-lysyl-proline (AcSDKP), an ACE N-domain specific substrate, mediates the protective phenotype of NKO. For this, diabetic mice were treated with the prolyl oligopeptidase inhibitor, S17092 (10 mg/kg, IP), that prevents the synthesis of AcSDKP. In diabetic NKO mice receiving S17092, sodium excretion in response to a saline challenge, ENaC α and γ subunit cleavage, renal inflammation and renal injury were indistinguishable from equally treated diabetic WT mice. In summary, these data indicate that increasing AcSDKP by blocking the ACE N-domain improves sodium handling and ameliorates diabetic kidney disease independently of intrarenal Ang II regulation.
Abstract 038: The Dual Role of B-Cell Lymphoma/Leukemia 10 in Angiotensin II-Induced Renal Damage
Authors: Markó Lajos , Park Joon-Keun , Henke Norbert , Rong Song , Balogh András , Klamer Samuel , Wilck Nicola , Ruland Jürgen , Luft Friedrich C , Dechend Ralf , Müller Dominik N
Published: December 2018

Angiotensin (Ang) II activates NF-κB and thereby induces organ damage. B-cell Lymphoma/Leukemia 10 (Bcl10) is a member of the CARMA-Bcl10-MALT1 signalosome that links Ang II and antigen-dependent activation of immune cells to NF-κB signaling. We have shown earlier that Bcl10 plays a significant role in the generation of Ang II–induced cardiac fibrosis and electric remodeling in a blood pressure independent fashion. In our present study we investigated the role of Bcl10 in Ang II–induced renal damage. Bcl10 knockout mice (Bcl10 KO) and wild-type (WT) controls were given 1% NaCl in the drinking water and Ang II (1.44 mg/kg/d) for 14 days. Additionally, Bcl10 KO or WT kidneys were transplanted onto WT mice and were challenged by the same protocol for 7 days. Kidneys of Bcl10 KO mice developed less fibrosis and showed reduced number of infiltrating macrophages and T cells. Nevertheless, renal expression of neutrophil gelatinase-associated lipocalin (Ngal) and kidney injury molecule 1 expression was higher after Ang II treatment in Bcl10 KO mice compared to WT mice indicating exacerbated tubular damage (4.94±1.56 vs 1.44±0.44 and 1.14±0.22 vs 2.87±1.05 AU respectively). In addition, albuminuria was significantly higher in Ang II-treated Bcl10 KO mice than in controls (1194±238 vs. 377±112 μg/day) accompanied by reduced glomerular nephrin expression and reduced podocyte number (5.9±0.4 vs. 8.2±0.5 Wilms’ tumor-1 protein (WT1) positive cells/glomerulus). Ang II-treated WT mice transplanted with Bcl10 KO kidney showed significantly higher albuminuria (6003±1988 vs. 1779±831 μg/day) and renal Ngal expression (1.75±4.08 vs 0.71±0.57 AU) compared to WT->WT kidney transplanted mice, as well as lower podocyte number (5.4±0.3 vs. 4.9±0.2 WT1 positive cells/glomerulus) but similar extent of fibrosis and cell infiltration. Lacking Bcl10 in the kidney only was protective from Ang II-induced cardiac hypertrophy. Bcl10 has a multi-faceted action in hypertensive renal damage. While global Bcl10 deficiency ameliorates renal fibrosis and cell infiltration, the lack of renal Bcl10 aggravates albuminuria and podocyte damage. These data indicate that Bcl10 plays a crucial role in maintaining podocyte integrity and renal function.
Abstract 047: Sorting Nexin 19: A Novel & Key Player in Renal Dopamine D
1
R Regulation
Authors: Tiu Andrew C , Yang Jian , Asico Laureano D , Konkalmatt Prasad , Wang Xiaoyan , Jose Pedro A , Villar Van Anthony M
Published: December 2018

The cellular localization and signal transduction of a ligand-occupied G protein-coupled receptor (GPCR) are tightly regulated to generate an appropriate response in terms of specificity, magnitude, and duration through the interplay of key proteins. We recently identified sorting nexin 19 (SNX19) as a binding partner for the renal dopamine D
1
receptor (D
1
R) and demonstrated its role in receptor lipid raft distribution and activity. We have shown that SNX19 and D
1
R co-immunoprecipitated (co-IP’d) and colocalized at the plasma membrane of human renal proximal tubule cells (hRPTCs) and at the brush border of RPTs of C57Bl/6 mice. Treatment with the D
1
R/D
5
R agonist fenoldopam promoted the colocalization to the cytoplasm of RPTCs. SNX19 silencing in hRPTCs decreased the D
1
R expression (-60±4% of basal, P<0.05, n=4) and impaired D
1
R recruitment and endocytosis following agonist stimulation, as observed via live cell imaging. SNX19 silencing markedly decreased the fenoldopam-mediated increase in cAMP production in hRPTCs (+115±30%
vs
. +45±12% in mock siRNA-treated control cells, P<0.05, n=4), and abrogated the fenoldopam-mediated inhibition of Na
+
,K
+
-ATPase activity in polarized hRPTCs (+21±1.2% intracellular Na
+
vs
. +2±2% in control cells, P<0.05, n=4). Kidney-restricted
Snx19
silencing in C57Bl/6 mice increased the systolic blood pressure (124±3 mm Hg
vs
. 101±2 mm Hg in control mice, P>0.05, n=5), thus demonstrating the importance of SNX19 in renal D
1
R activity and blood pressure control. SNX19 also co-IP’d with GRK4 and GODZ finger protein, an enzyme involved in GPCR palmitoylation, which is a process that promotes lipid raft partitioning. Mutation of the canonical PX domain of SNX19, which prevented its targeting to the plasma membrane, and the heterologous expression of these mutants in hRPTCs decreased the abundance of D
1
R, which became exclusively distributed in non-lipid rafts. The disruption of lipid rafts in the kidneys of mice resulted in hypertension (120±3 mm Hg
vs
. 101±2 mm Hg in control mice, n=4). Our results highlight the crucial role of SNX19 in the proper trafficking and functioning of renal D
1
R through its ability to promote the localization of D
1
R in lipid rafts for an effective signal transduction and appropriate cellular response.