bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin 10 Gene-Modified Bone Marrow-Derived Dendritic Cells Attenuate Liver Fibrosis in Mice by Inducing Regulatory T Cells and Inhibiting the TGF-β/Smad Signaling Pathway
Authors: Xu Yejin , Tang Xinyue , Yang Min , Zhang Shengguo , Li Shanshan , Chen Yukai , Liu Minhui , Guo Yuxiang , Lu Mingqin
Published: January 2019

Aim. To explore the therapeutic effects and mechanisms of interleukin 10 gene-modified bone marrow-derived dendritic cells (DC-IL10) on liver fibrosis. Methods. In vitro, BMDCs were transfected with lentiviral-interleukin 10-GFP (LV-IL10-GFP) at the MOI of 1 : 40. Then, the phenotype (MHCII, CD80, and CD86) and allo-stimulatory ability of DC-IL10 were identified by flow cytometry, and the levels of IL-10 and IL-12 (p70) secreted into the culture supernatants were quantified by ELISA. In vivo, DC-IL10 was injected into mice with CCl4-induced liver fibrosis through the tail vein. Lymphocytes were isolated to investigate the differentiation of T cells, and serum and liver tissue were collected for biochemical, cytokine, histopathologic, immune-histochemical, and Western blot analyzes. Results. In vitro, the expressions of MHCII, CD80, and CD86 in DC-IL10 were significantly suppressed, allogeneic CD4+T cells incubated with DC-IL10 showed a lower proliferative response, and the levels of IL-10 and IL-12 (p70) secreted into the DC-IL10 culture supernatants were significantly increased and decreased, respectively. In vivo, regulatory T cells (Tregs) were significantly increased, while ALT, AST, and inflammatory cytokines were significantly reduced in the DC-IL10 treatment group, and the degree of hepatic fibrosis was obviously reversed. The TGF-β/smad pathway was inhibited following DC-IL10 treatment compared to the liver fibrosis group. Conclusion. IL-10 genetic modification of BMDCs may maintain DC in the state of tolerance and allow DC to induce T cell hyporesponsiveness or tolerance. DC-IL10 suppressed liver fibrosis by inducing Treg production and inhibiting the TGF-β/smad signaling pathway.
Factors Associated with Health-Related Quality of Life among Jordanian Patients with Diabetic Foot Ulcer
Authors: Alrub Ahmad Abu , Hyassat Dana , Khader Yousef S. , Bani-Mustafa Radwan , Younes Nidal , Ajlouni Kamel
Published: January 2019

Objective. This study is aimed at determining factors associated with the quality of life among Jordanian diabetic patients with foot ulcers. Methods. 144 consecutive patients with diabetic foot ulcers aged ≥ 18 years who were attending the diabetic foot clinic at a diabetes-specialized center were included in this study. Health-related quality of life was assessed using two self-administered questionnaires: Diabetic Foot Scale-Short Form (DFS-SF) and Short Form-8 (SF-8). Results. Patients with diabetic foot ulcer had low mean DFS-SF score and low mean scores on physical and mental component summary scales (PCS8 and MCS8). Males had significantly higher DFS-SF score indicating better health-related quality of life than females (P value 0.038). A patient with stressful life events had significantly lower health-related quality of life using DFS-SF scale and SF-8 summary scales. Patients with peripheral vascular disease (PVD) and patients with obesity had lower DFS-SF and PCS8 quality of life. Conclusion. Patients with diabetic foot ulcer had low quality of life. Female gender, obesity, presence of PVD, and stressful life events were the most important factors associated with lower quality of life in patients with diabetic foot ulcer.
Code Division Multiplexing and Machine Learning Based Reversible Data Hiding Scheme for Medical Image
Authors: Ma Bin , Li Bing , Wang Xiao-Yu , Wang Chun-Peng , Li Jian , Shi Yun-Qing
Published: January 2019

In this paper, a new reversible data hiding (RDH) scheme based on Code Division Multiplexing (CDM) and machine learning algorithms for medical image is proposed. The original medical image is firstly converted into frequency domain with integer-to-integer wavelet transform (IWT) algorithm, and then the secret data are embedded into the medium frequency subbands of medical image robustly with CDM and machine learning algorithms. According to the orthogonality of different spreading sequences employed in CDM algorithm, the secret data are embedded repeatedly, most of the elements of spreading sequences are mutually canceled, and the proposed method obtained high data embedding capacity at low image distortion. Simultaneously, the to-be-embedded secret data are represented by different spreading sequences, and only the receiver who has the spreading sequences the same as the sender can extract the secret data and original image completely, by which the security of the RDH is improved effectively. Experimental results show the feasibility of the proposed scheme for data embedding in medical image comparing with other state-of-the-art methods.
Magnetic Resonance Angiography Shows Increased Arterial Blood Supply Associated with Murine Mammary Cancer
Authors: Mustafi Devkumar , Leinroth Abby , Fan Xiaobing , Markiewicz Erica , Zamora Marta , Mueller Jeffrey , Conzen Suzanne D. , Karczmar Gregory S.
Published: January 2019

Breast cancer is a major cause of morbidity and mortality in Western women. Tumor neoangiogenesis, the formation of new blood vessels from pre-existing ones, may be used as a prognostic marker for cancer progression. Clinical practice uses dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) to detect cancers based on increased blood flow and capillary permeability. However, DCE-MRI requires repeated injections of contrast media. Therefore we explored the use of noninvasive time-of-flight (TOF) MR angiography for serial studies of mouse mammary glands to measure the number and size of arteries feeding mammary glands with and without cancer. Virgin female C3(1) SV40 TAg mice (n=9), aged 18-20 weeks, were imaged on a 9.4 Tesla small animal scanner. Multislice T2-weighted (T2W) images and TOF-MRI angiograms were acquired over inguinal mouse mammary glands. The data were analyzed to determine tumor burden in each mammary gland and the volume of arteries feeding each mammary gland. After in vivo MRI, inguinal mammary glands were excised and fixed in formalin for histology. TOF angiography detected arteries with a diameter as small as 0.1 mm feeding the mammary glands. A significant correlation (r=0.79; p< 0.0001) was found between tumor volume and the arterial blood volume measured in mammary glands. Mammary arterial blood volumes ranging from 0.08 mm3 to 3.81 mm3 were measured. Tumors and blood vessels found on in vivo T2W and TOF images, respectively, were confirmed with ex vivo histological images. These results demonstrate increased recruitment of arteries to mammary glands with cancer, likely associated with neoangiogenesis. Neoangiogenesis may be detected by TOF angiography without injection of contrast agents. This would be very useful in mouse models where repeat placement of I.V. lines is challenging. In addition, analogous methods could be tested in humans to evaluate the vasculature of suspicious lesions without using contrast agents.
Chlamydia trachomatis Growth and Cytokine mRNA Response in a Prostate Cancer Cell Line
Authors: Petyaev Ivan M. , Zigangirova Naylia A. , Morgunova Elena Y. , Kyle Nigel H. , Fedina Elena D. , Bashmakov Yuriy K.
Published: January 2019

In the present paper, we report that C. trachomatis can be efficiently propagated and affect mRNA expression for two major cytokines, relevant to tumor progression, in CWR-R1 cells, a malignant prostate cell line. CWR-R1 and McCoy cells, a classic cell line for chlamydial research, were grown and infected with C. trachomatis under similar conditions. Cell monolayers were harvested for RNA analysis and immunostaining with major outer membrane protein (MOMP) antibody at 24, 48, and 72 hours of the postinfection (hpi) period. It was shown that the infectious cycle of chlamydial pathogen in CWR-R1 cells resembles the progression of C. trachomatis infection in McCoy cells but with a few important differences. First of all, the initial stage of C. trachomatis propagation in CWR-R1 cells (24 hpi) was characterized by larger inclusion bodies and more intense, specific immunofluorescent staining of infected cells as compared with McCoy cells. Moreover, there was a corresponding increase in infective progeny formation in CWR-R1 cells along with mRNA for EUO, a crucial gene controlling the early phase of the chlamydial development cycle (24 hpi). These changes were more minimal and became statistically insignificant at a later time point in the infectious cycle (48 hpi). Altogether, these data suggest that the early phase of C. trachomatis infection in CWR-R1 cells is accompanied by more efficient propagation of the pathogen as compared with the growth of C. trachomatis in McCoy cells. Furthermore, propagation of C. trachomatis in CWR-R1 cells leads to enhanced transcription of interleukin-6 and fibroblast growth factor-2, genes encoding two important proinflammatory cytokines implicated in the molecular mechanisms of chemoresistance of prostate cancer and its ability to metastasize. The possible roles of reactive oxygen species and impaired mitochondrial oxidation in the prostate cancer cell line are discussed as factors promoting the early stages of C. trachomatis growth in CWR-R1 cells.
Dynamic Parameter Analysis of Spindle Bearing Using 3‐Dimension Quasi‐Dynamic Model
Authors: Yu Jie , Li Songsheng , Chen Xiaoyang , He Lei , Yuan Wei
Published: January 2019

Spindle bearing with high rotational speed is the kernel of many rotating machines which plays massive important role in the performance of the whole machine. There are also some deficiencies of the present quasi‐dynamic mathematical models of the rolling bearing in analyzing the bearing dynamic parameters. This paper develops the 3D quasi‐dynamic model with ball’s 6‐DOF, cage’s 6‐DOF, and driving ring’s 5‐DOF under steady working stage, based on the analysis of the relative position of each component to determine the normal forces between each component and the analysis of the relative velocity of the contacting point to determine the tangential force between each component. This model can analyze the dynamic parameters, such as the spin‐to‐roll ratio of inner and outer rings, the angular velocity of rolling elements in y‐axis of the location coordinate, the normal contact force of cage pocket and ball, the cage angular velocity and ball revolution velocity, and the gyroscopic moment under different working condition. Compared with the numerical calculation results of ADORE program, the results of the proposed model are able to accurately describe the change laws of the spindle bearing parameters.
Study on Multiparameter Precursory Information Identification of the Fracture of Yellow Sandstone
Authors: Fukun Xiao , Houran Wang , Gang Liu
Published: January 2019

In order to explore the disaster caused by uncontrollable instability of coal and rock mass, a multiparameter fusion system is constructed to predict and predict disasters more accurately by identifying the mechanical and acoustic precursors of coal and rock fracture. In order to explore the precursor information of yellow sandstone rupture, the damage evolution process of yellow sandstone is analyzed from the four aspects of rock mechanics, acoustic emission time domain, frequency domain, and characteristic parameters, and the body strain, dissipated energy and acoustic emission counting, acoustic emission energy, average frequency, peak frequency, b value, and entropy value precursor information identification points are obtained, and 8 parameters are analyzed by time series fusion. The specific conclusions are as follows: body strain in the violent stage of damage evolution, the slope is zero, the zero end point is the precursor information identification point, the dissipative energy curve overall shows the “s” type, the early growth rate is faster—the medium‐term stability—the later period is slowed down, and the upper slope boundary point of the “s” type curve is used as the precursor information identification point. In the violent stage of damage evolution, the layered features of the acoustic emission count are obvious, the specific gravity shift is more obvious, and the high count appears as the precursor information identification point; the acoustic emission energy accumulates the high‐energy signal and is accompanied by the steady and rapid growth of energy as the precursor information identification point. The effects of shearing main cracks, shear microcracks, tensile cracks, and composite cracks on the acoustic emission count and energy in the damage evolution process are analyzed. The increase of medium‐ and high‐frequency signals and the reduction of high‐frequency signals predict the rupture. The average frequency signal change law is continuous high frequency‐blank‐continuous high frequency, with the blank period end point as the damage precursor identification point; the b value damage evolution stage shows a continuously steady increase to a rapid increase, with the continuous stable growth starting point as the crack identification point. In the process of damage evolution, the sample entropy presents an orderly, chaotic, disordered, and orderly process. The end of chaos and the beginning of disorder are used as the prejudging demarcation points. Based on the time sequence, an eight‐parameter comprehensive early warning system is constructed. The indicators are classified into five levels for early warning in the stage of severe damage evolution. The identification of multiparameter precursory information of yellow sandstone provides a new research idea and analysis angle and method for the failure of other coal and rock masses.
Influence of Age on Anticontractile Effect of Perivascular Adipose Tissue in Normotensive and Hypertensive Rats
Authors: Zemančíková Anna , Török Jozef
Published: January 2019

Perivascular adipose tissue (PVAT) and its vasomodulatory effects play an important role in the physiology and pathophysiology of blood vessels. Alterations in PVAT associated with reduction in its anticontractile influence are proven to contribute to vascular dysfunction in hypertension. The aim of this study was to examine whether the changes in PVAT properties could participate in progression of vascular abnormalities in developing spontaneously hypertensive rats (SHR). Normotensive Wistar-Kyoto (WKY) rats and SHR, both in 5th and in 12th week of age, were used. Systolic blood pressure was similar between WKY rats and SHR in 5th week of age; however, in 12th week, it was significantly increased in SHR comparing to WKY rats. The amount of retroperitoneal fat was higher in WKY rats in both age groups, whereas body weight was higher in WKY rats only in 12th week, when compared to age-matched SHR. From isolated superior mesenteric arteries, two ring preparations were prepared for isometric tension recording, one with PVAT intact and other with PVAT removed. In WKY rats as well as in SHR, arterial contractile responses to noradrenaline, applied cumulatively on rings, were significantly inhibited in the presence of intact PVAT. In both age groups, anticontractile effect of PVAT was higher in WKY rats than in SHR. Neurogenic contractions, induced by electrical stimulation of perivascular sympathoadrenergic nerves, were significantly attenuated in the presence of PVAT in WKY mesenteric arteries from both age groups; however, in arteries from SHR, intact PVAT had no influence on this type of contractile responses. The results suggest that in SHR impairment of anticontractile effect of PVAT precedes hypertension and might contribute to its development.
CHIP: Clustering Hotspots in Layout Using Integer Programming
Authors: Takkala Rohit Reddy , Chu Chris
Published: January 2019

Clustering algorithms have been explored in recent years to solve hotspot clustering problems in integrated circuit design. With various applications in design for manufacturability flow such as hotspot library generation, systematic yield optimization, and design space exploration, generating good quality clusters along with their representative clips is of utmost importance. With several generic clustering algorithms at our disposal, hotspots can be clustered based on the distance metric defined while satisfying some tolerance conditions. However, the clusters generated from generic clustering algorithms need not achieve optimal results. In this paper, we introduce two optimal integer linear programming formulations based on triangle inequality to solve the problem of minimizing cluster count while satisfying given constraints. Apart from minimizing cluster count, we generate representative clips that best represent the clusters formed. We achieve a better cluster count for both formulations in most test cases as compared to the results published in the literature in the ICCAD 2016 contest benchmarks as well as the reference results reported in the ICCAD 2016 contest website.
An Approach for Pulmonary Vascular Extraction from Chest CT Images
Authors: Tan Wenjun , Yuan Yue , Chen Anning , Mao Lin , Ke Yuqian , Lv Xinhui
Published: January 2019

Pulmonary vascular extraction from chest CT images plays an important role in the diagnosis of lung disease. To improve the accuracy rate of pulmonary vascular segmentation, a new pulmonary vascular extraction approach is proposed in this study. First, the lung tissue is extracted from chest CT images by region-growing and maximum between-class variance methods. Then the holes of the extracted region are filled by morphological operations to obtain complete lung region. Second, the points of the pulmonary vascular of the middle slice of the chest CT images are extracted as the original seed points. Finally, the seed points are spread throughout the lung region based on the fast marching method to extract the pulmonary vascular in the gradient image. Results of pulmonary vascular extraction from chest CT image datasets provided by the introduced approach are presented and discussed. Based on the ground truth pixels and the resulting quality measures, it can be concluded that the average accuracy of this approach is about 90%. Extensive experiments demonstrate that the proposed method has achieved the best performance in pulmonary vascular extraction compared with other two widely used methods.
CAR T Cells Targeting B7-H3, a Pan-Cancer Antigen, Demonstrate Potent Preclinical Activity Against Pediatric Solid Tumors and Brain Tumors
Authors: Majzner Robbie G. , Theruvath Johanna L. , Nellan Anandani , Heitzeneder Sabine , Cui Yongzhi , Mount Christopher W. , Rietberg Skyler P. , Linde Miles H. , Xu Peng , Rota Christopher , Sotillo Elena , Labanieh Louai , Lee Daniel W. , Orentas Rimas J. , Dimitrov Dimiter S. , Zhu Zhongyu , Croix Brad St , Delaidelli Alberto , Sekunova Alla , Bonvini Ezio , Mitra Siddhartha S. , Quezado Martha M. , Majeti Ravindra , Monje Michelle , Sorensen Poul H.B. , Maris John M. , Mackall Crystal L.
Published: January 2019

Purpose:

Patients with relapsed pediatric solid tumors and CNS malignancies have few therapeutic options and frequently die of their disease. Chimeric antigen receptor (CAR) T cells have shown tremendous success in treating relapsed pediatric acute lymphoblastic leukemia, but this has not yet translated to treating solid tumors. This is partially due to a paucity of differentially expressed cell surface molecules on solid tumors that can be safely targeted. Here, we present B7-H3 (CD276) as a putative target for CAR T-cell therapy of pediatric solid tumors, including those arising in the central nervous system.

Experimental Design:

We developed a novel B7-H3 CAR whose binder is derived from a mAb that has been shown to preferentially bind tumor tissues and has been safely used in humans in early-phase clinical trials. We tested B7-H3 CAR T cells in a variety of pediatric cancer models.

Results:

B7-H3 CAR T cells mediate significant antitumor activity in vivo, causing regression of established solid tumors in xenograft models including osteosarcoma, medulloblastoma, and Ewing sarcoma. We demonstrate that B7-H3 CAR T-cell efficacy is largely dependent upon high surface target antigen density on tumor tissues and that activity is greatly diminished against target cells that express low levels of antigen, thus providing a possible therapeutic window despite low-level normal tissue expression of B7-H3.

Conclusions:

B7-H3 CAR T cells could represent an exciting therapeutic option for patients with certain lethal relapsed or refractory pediatric malignancies, and should be tested in carefully designed clinical trials.
The RUNX Transcriptional Coregulator, CBFβ, Suppresses Migration of ER+ Breast Cancer Cells by Repressing ERα-Mediated Expression of the Migratory Factor TFF1
Authors: Pegg Henry J. , Harrison Hannah , Rogerson Connor , Shore Paul
Published: January 2019

Core binding factor β (CBFβ), the essential coregulator of RUNX transcription factors, is one of the most frequently mutated genes in estrogen receptor–positive (ER+) breast cancer. Many of these mutations are nonsense mutations and are predicted to result in loss of function, suggesting a tumor suppressor role for CBFβ. However, the impact of missense mutations and the loss of CBFβ in ER+ breast cancer cells have not been determined. Here we demonstrate that missense mutations in CBFβ accumulate near the Runt domain–binding region. These mutations inhibit the ability of CBFβ to form CBFβ–Runx–DNA complexes. We further show that deletion of CBFβ, using CRISPR-Cas9, in ER+ MCF7 cells results in an increase in cell migration. This increase in migration is dependent on the presence of ERα. Analysis of the potential mechanism revealed that the increase in migration is driven by the coregulation of Trefoil factor 1 (TFF1) by CBFβ and ERα. RUNX1–CBFβ acts to repress ERα-activated expression of TFF1. TFF1 is a motogen that stimulates migration and we show that knockdown of TFF1 in CBFβ−/− cells inhibits the migratory phenotype. Our findings reveal a new mechanism by which RUNX1–CBFβ and ERα combine to regulate gene expression and a new role for RUNX1–CBFβ in the prevention of cell migration by suppressing the expression of the motogen TFF1.

Implications:

Mutations in CBFβ contribute to the development of breast cancer by inducing a metastatic phenotype that is dependent on ER.
Efficacy and Safety of Topical 3% Diquafosol Ophthalmic Solution for the Treatment of Multifactorial Dry Eye Disease: Meta-Analysis of Randomized Clinical Trials
Authors: Nam Kiyeun , Kim Hyun Jung , Yoo Aeri
Published: January 2019

Purpose: A meta-analysis was performed to evaluate the safety and efficacy of topical 3% diquafosol in treating patients with dry eye disease (DED). Methods: Nine qualified randomized controlled trials incorporating 1,467 patients were included. Two of the reviewers selected the studies and independently assessed the risk of bias. The outcome measures were Schirmer score, tear film break-up time (TFBUT), rose bengal staining score, and corneal fluorescein staining score. To confirm the effect of diquafosol on dryness after cataract surgery, we performed a subgroup analysis according to the presence or absence of surgery. Results: We observed statistically significant improvements in scores on the Schirmer test (weighted mean difference 0.74 mm at 4 weeks; 95% CI: 0.24–1.24; I2 = 0%), fluorescein stain, rose bengal stain, and TFBUT after treatment with diquafosol compared with the group using other eye drops. As a result of the subgroup analysis of DED after cataract surgery, diquafosol was found to be more effective than the other eye drops with regard to TFBUT and rose bengal staining. Conclusions: Topical diquafosol could be an effective treatment for DED, and also for DED after cataract surgery. Further randomized controlled trials with larger sample sizes for the different clinical types of DED are warranted to determine the efficacy and limitations of diquafosol for these different clinical types of DED.
Chondrosarcoma Metastasis to the Choroid
Authors: Kim Jenna May , Wong Amanda J. , Lu Amanda J. , Pointdujour-Lim Renelle
Published: January 2019

We report a rare case of chondrosarcoma metastatic to the choroid. A 64-year-old male with a history of chondrosarcoma metastatic to the lungs and to the spine presented with blurred vision. A choroidal tumor was found. Fine-needle biopsy confirmed the histologic identity of the tumor as chondrosarcoma. Metastatic spread of chondrosarcoma to the eye is extremely rare. When present, lesions may grow rapidly, and systemic prognosis is poor. Co-management with medical oncology is of utmost importance. This is the third case of chondrosarcoma metastatic to the choroid in the literature and the first with bilateral involvement.
Malnutrition Risk in Hemodialysis Patients in Israel: Results of the Status of Nutrition In Hemodialysis Patients Survey Study
Authors: Boaz Mona , Azoulay Odile , Schwartz Idit F. , Schwartz Doron , Assady Suhair , Kristal Batya , Benshitrit Sydney , Yanai Noa , Weinstein Talia
Published: January 2019

Background: Malnutrition is a frequently observed disorder in patients with chronic kidney disease (CKD) receiving hemodialysis (HD). While variously defined, malnutrition is a frequently observed condition among patients on HD. Prevalence estimates of malnutrition among Israeli HD patients have not been reported. Objectives: To survey nutrition intake in Israeli HD patients; estimate malnutrition risk prevalence; identify risk factors, and characterize malnutrition risk in HD patients. Methods: A representative sample of 378 Israeli HD patients treated in hospital HD centers throughout the country were surveyed. Using the 24-h recall method, dietary intake was estimated and the chronologically corresponding biochemistry, anthropometric, and hemodynamic measures were recorded. Four categories of malnutrition risk were defined: “minimal”: body mass index (BMI) > 23 kg/m2 and serum albumin > 3.8 g/dL; “mild”: BMI < 23 kg/m2 and albumin > 3.8 g/dL; “moderate”: BMI > 23 kg/m2 and albumin < 3.8 g/dL; “severe”: BMI < 23 k/m2 and serum albumin < 3.8 g/dL. Results: Elevated malnutrition risk was identified in 175 (46.3%) study participants, who were more likely to require feeding assistance, have major comorbidities, reduced hemoglobin, and elevated C-reactive protein. Oral nutrition supplementation was prescribed to only 14.3% of patients with elevated malnutrition risk. Intradialytic parenteral nutrition was recorded for 6 patients, all of whom had moderate or severe malnutrition risk. Less than one-third of patients met the guidelines for dietary intake of energy or protein, and this did not differ across malnutrition risk groups. Discussion: Elevated malnutrition risk is a frequent finding in HD patients treated in hospital settings in Israel. Dietary intake does not meet guidelines but does not differ across malnutrition risk categories. Nutrition supplements are underused in HD patients with high malnutrition risk. There is a need to expand the survey to community HD centers.
Endoscopic Retrieval of Mis-Swallowed Table Spoon
Authors: Shen Chun-Sheng , Su Yu-Chung
Published: January 2019

Mis-swallowed foreign body is a common scenario for endoscopy intervention. Although a long object is rarely seen in our clinical setting, limited published articles could be found for proper retrieval. We hence report a 30-year-old female who mis-swallowed a long table spoon, which was retrieved by a snare from the duodenal second portion with conventional endoscopy.
Admission Levels of DKK1 (Dickkopf-1) Are Associated With Future Cardiovascular Death in Patients With Acute Coronary Syndromes
Authors: Ueland Thor , Åkerblom Axel , Ghukasyan Tatevik , Michelsen Annika E. , Becker Richard C. , Bertilsson Maria , Himmelmann Anders , James Stefan K. , Siegbahn Agneta , Storey Robert F. , Kontny Frederic , Aukrust Pål , Wallentin Lars ,   for the PLATO Investigators
Published: January 2019

Objective—

The Wnt/wingless signaling antagonist DKK1 (dickkopf-1) regulates platelet-mediated inflammation and may contribute to plaque destabilization. We hypothesized that DKK1 would be associated with cardiovascular outcomes.

Approach and Results—

We determined DKK1 levels in serum samples obtained before randomization, at discharge, and 1 and 6 months in a subset of 5165 patients with acute coronary syndromes in the PLATO trial (Platelet Inhibition and Patient Outcomes; NCT00391872). The median (interquartile range) DKK1 concentrations were 0.61 (0.20–1.27) ng/mL at baseline and increased during follow-up. The hazard ratio (95% CIs) for the composite end point (cardiovascular death, nonprocedural spontaneous myocardial infarction, or stroke) during 1 year of follow-up, per 50% increase in baseline DKK1 concentration, was 1.06 (1.02–1.10),
P
=0.0011, and remained significant in fully adjusted analysis with 14 conventional clinical and demographic and 6 biochemical variables, including NT-proBNP (N-terminal pro-B-type natriuretic peptide), hs-TnT (high-sensitivity troponin T), and GDF-15 (growth differentiation factor 15; 1.05 [1.00–1.09];
P
=0.028). This association was mainly driven by the association with cardiovascular death, where a gradual increase in event rates was observed with increasing quartiles of DKK1 (2.7%, 3.0%, 4.3%, and 5.0%) and remained significant and unmodified in fully adjusted analysis (hazard ratio, 1.10 [1.04–1.17];
P
=0.002). Change in DKK1 and levels at 1 month were unrelated to outcomes. A modifying effect of ticagrelor on DKK1 discharge levels was observed but not associated with prognosis.

Conclusions—

In patients with acute coronary syndromes treated with dual antiplatelet treatment, admission DKK1 levels were independently associated with a composite of cardiovascular death, myocardial infarction, or stroke and with cardiovascular death alone.
Amino Acids and Their Metabolism in Atherosclerosis
Authors: Nitz Katrin , Lacy Michael , Atzler Dorothee
Published: January 2019

As a leading cause of death worldwide, cardiovascular disease is a global health concern. The development and progression of atherosclerosis, which ultimately gives rise to cardiovascular disease, has been causally linked to hypercholesterolemia. Mechanistically, the interplay between lipids and the immune system during plaque progression significantly contributes to the chronic inflammation seen in the arterial wall during atherosclerosis. Localized inflammation and increased cell-to-cell interactions may influence polarization and proliferation of immune cells via changes in amino acid metabolism. Specifically, the amino acids
l
-arginine (Arg),
l
-homoarginine (hArg) and
l
-tryptophan (Trp) have been widely studied in the context of cardiovascular disease, and their metabolism has been established as key regulators of vascular homeostasis, as well as immune cell function. Cyclic effects between endothelial cells, innate, and adaptive immune cells exist during Arg and hArg, as well as Trp metabolism, that may have distinct effects on the development of atherosclerosis. In this review, we describe the current knowledge surrounding the metabolism, biological function, and clinical perspective of Arg, hArg, and Trp in the context of atherosclerosis.
CD69 Plays a Beneficial Role in Ischemic Stroke by Dampening Endothelial Activation
Authors: Brait Vanessa H. , Miró-Mur Francesc , Pérez-de-Puig Isabel , Notario Laura , Hurtado Begoña , Pedragosa Jordi , Gallizioli Mattia , Jiménez-Altayó Francesc , Arbaizar-Rovirosa Maria , Otxoa-de-Amezaga Amaia , Monteagudo Juan , Ferrer-Ferrer Maura , de la Rosa Xavier , Bonfill-Teixidor Ester , Salas-Perdomo Angélica , Hernández-Vidal Alba , Garcia-de-Frutos Pablo , Lauzurica Pilar , Planas Anna M.
Published: January 2019

Rationale:

CD69 is an immunomodulatory molecule induced during lymphocyte activation. Following stroke, T-lymphocytes upregulate CD69 but its function is unknown.

Objective:

We investigated whether CD69 was involved in brain damage following an ischemic stroke.

Methods and Results:

We used adult male mice on the C57BL/6 or BALB/c backgrounds, including wild-type mice and CD69
−/−
mice, and CD69
+/+
and CD69
−/−
lymphocyte-deficient Rag2
−/
−
mice, and generated chimeric mice. We induced ischemia by transient or permanent middle cerebral artery occlusion. We measured infarct volume, assessed neurological function, and studied CD69 expression, as well as platelet function, fibrin(ogen) deposition, and VWF (von Willebrand factor) expression in brain vessels and VWF content and activity in plasma, and performed the tail-vein bleeding test and the carotid artery ferric chloride-induced thrombosis model. We also performed primary glial cell cultures and sorted brain CD45
−
CD11b
−
CD31
+
endothelial cells for mRNA expression studies. We blocked VWF by intravenous administration of anti-VWF antibodies. CD69
−/−
mice showed larger infarct volumes and worse neurological deficits than the wild-type mice after ischemia. This worsening effect was not attributable to lymphocytes or other hematopoietic cells. CD69 deficiency lowered the time to thrombosis in the carotid artery despite platelet function not being affected. Ischemia upregulated
Cd69
mRNA expression in brain endothelial cells. CD69-deficiency increased fibrin(ogen) accumulation in the ischemic tissue, and plasma VWF content and activity, and VWF expression in brain vessels. Blocking VWF reduced infarct volume and reverted the detrimental effect of CD69
−/−
deficiency.

Conclusions:

CD69 deficiency promotes a prothrombotic phenotype characterized by increased VWF and worse brain damage after ischemic stroke. The results suggest that CD69 acts as a downregulator of endothelial activation.
Bmpr2
Mutant Rats Develop Pulmonary and Cardiac Characteristics of Pulmonary Arterial Hypertension
Authors: Hautefort Aurélie , Mendes-Ferreira Pedro , Sabourin Jessica , Manaud Grégoire , Bertero Thomas , Rucker-Martin Catherine , Riou Marianne , Adão Rui , Manoury Boris , Lambert Mélanie , Boet Angèle , Lecerf Florence , Domergue Valérie , Brás-Silva Carmen , Gomez Ana Maria , Montani David , Girerd Barbara , Humbert Marc , Antigny Fabrice , Perros Frédéric
Published: January 2019

Background:

Monoallelic mutations in the gene encoding bone morphogenetic protein receptor 2 (
Bmpr2
) are the main genetic risk factor for heritable pulmonary arterial hypertension (PAH) with incomplete penetrance. Several
Bmpr2
transgenic mice have been reported to develop mild spontaneous PAH. In this study, we examined whether rats with the
Bmpr2
mutation were susceptible to developing more severe PAH.

Methods:

The zinc finger nuclease method was used to establish rat lines with mutations in the
Bmpr2
gene. These rats were then characterized at the hemodynamic, histological, electrophysiological, and molecular levels.

Results:

Rats with a monoallelic deletion of 71 bp in exon 1 (Δ
71
rats) showed decreased BMPRII expression and phosphorylated SMAD1/5/9 levels. Δ
71
Rats develop age-dependent spontaneous PAH with a low penetrance (16%–27%), similar to that in humans. Δ
71
Rats were more susceptible to hypoxia-induced pulmonary hypertension than wild-type rats. Δ
71
Rats exhibited progressive pulmonary vascular remodeling associated with a proproliferative phenotype and showed lower pulmonary microvascular density than wild-type rats. Organ bath studies revealed severe alteration of pulmonary artery contraction and relaxation associated with potassium channel subfamily K member 3 (KCNK3) dysfunction. High levels of perivascular fibrillar collagen and pulmonary interleukin-6 overexpression discriminated rats that developed spontaneous PAH and rats that did not develop spontaneous PAH. Finally, detailed assessments of cardiomyocytes demonstrated alterations in morphology, calcium (Ca
2+
), and cell contractility specific to the right ventricle; these changes could explain the lower cardiac output of Δ
71
rats. Indeed, adult right ventricular cardiomyocytes from Δ
71
rats exhibited a smaller diameter, decreased sensitivity of sarcomeres to Ca
2+
, decreased [Ca
2+
] transient amplitude, reduced sarcoplasmic reticulum Ca
2+
content, and short action potential duration compared with right ventricular cardiomyocytes from wild-type rats.

Conclusions:

We characterized the first
Bmpr2
mutant rats and showed some of the critical cellular and molecular dysfunctions described in human PAH. We also identified the heart as an unexpected but potential target organ of
Bmpr2
mutations. Thus, this new genetic rat model represents a promising tool to study the pathogenesis of PAH.
Phenotypic Refinement of Heart Failure in a National Biobank Facilitates Genetic Discovery
Authors: Aragam Krishna G. , Chaffin Mark , Levinson Rebecca T. , McDermott Gregory , Choi Seung Hoan , Shoemaker M. Benjamin , Haas Mary E. , Weng Lu-Chen , Lindsay Mark E. , Smith J. Gustav , Newton-Cheh Christopher , Roden Dan M. , London Barry ,   For the GRADE Investigators, Wells Quinn S. , Ellinor Patrick T. , Kathiresan Sekar , Lubitz Steven A. , Bloom Heather L. , Dudley Samuel C. , Shalaby Alaa A. , Weiss Raul , Gutmann Rebecca , Saba Samir
Published: January 2019

Background:

Heart failure (HF) is a morbid and heritable disorder for which the biological mechanisms are incompletely understood. We therefore examined genetic associations with HF in a large national biobank, and assessed whether refined phenotypic classification would facilitate genetic discovery.

Methods:

We defined all-cause HF among 488 010 participants from the UK Biobank and performed a genome-wide association analysis. We refined the HF phenotype by classifying individuals with left ventricular dysfunction and without coronary artery disease as having nonischemic cardiomyopathy (NICM), and repeated a genetic association analysis. We then pursued replication of lead HF and NICM variants in independent cohorts, and performed adjusted association analyses to assess whether identified genetic associations were mediated through clinical HF risk factors. In addition, we tested rare, loss-of-function mutations in 24 known dilated cardiomyopathy genes for association with HF and NICM. Finally, we examined associations between lead variants and left ventricular structure and function among individuals without HF using cardiac magnetic resonance imaging (n=4158) and echocardiographic data (n=30 201).

Results:

We identified 7382 participants with all-cause HF in the UK Biobank. Genome-wide association analysis of all-cause HF identified several suggestive loci (
P
<1×10
–6
), the majority linked to upstream HF risk factors, ie, coronary artery disease (
CDKN2B-AS1
and
MAP3K7CL
) and atrial fibrillation (
PITX2
). Refining the HF phenotype yielded a subset of 2038 NICM cases. In contrast to all-cause HF, genetic analysis of NICM revealed suggestive loci that have been implicated in dilated cardiomyopathy (
BAG3
,
CLCNKA-ZBTB17
). Dilated cardiomyopathy signals arising from our NICM analysis replicated in independent cohorts, persisted after HF risk factor adjustment, and were associated with indices of left ventricular dysfunction in individuals without clinical HF. In addition, analyses of loss-of-function variants implicated
BAG3
as a disease susceptibility gene for NICM (loss-of-function variant carrier frequency=0.01%; odds ratio,12.03;
P
=3.62×10
–5
).

Conclusions:

We found several distinct genetic mechanisms of all-cause HF in a national biobank that reflect well-known HF risk factors. Phenotypic refinement to a NICM subtype appeared to facilitate the discovery of genetic signals that act independently of clinical HF risk factors and that are associated with subclinical left ventricular dysfunction.
The N
6
-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy
Authors: Dorn Lisa E. , Lasman Lior , Chen Jing , Xu Xianyao , Hund Thomas J. , Medvedovic Mario , Hanna Jacob H. , van Berlo Jop H. , Accornero Federica
Published: January 2019

Background:

N
6
-Methyladenosine (m6A) methylation is the most prevalent internal posttranscriptional modification on mammalian mRNA. The role of m6A mRNA methylation in the heart is not known.

Methods:

To determine the role of m6A methylation in the heart, we isolated primary cardiomyocytes and performed m6A immunoprecipitation followed by RNA sequencing. We then generated genetic tools to modulate m6A levels in cardiomyocytes by manipulating the levels of the m6A RNA methylase methyltransferase-like 3 (METTL3) both in culture and in vivo. We generated cardiac-restricted gain- and loss-of-function mouse models to allow assessment of the METTL3-m6A pathway in cardiac homeostasis and function.

Results:

We measured the level of m6A methylation on cardiomyocyte mRNA, and found a significant increase in response to hypertrophic stimulation, suggesting a potential role for m6A methylation in the development of cardiomyocyte hypertrophy. Analysis of m6A methylation showed significant enrichment in genes that regulate kinases and intracellular signaling pathways. Inhibition of METTL3 completely abrogated the ability of cardiomyocytes to undergo hypertrophy when stimulated to grow, whereas increased expression of the m6A RNA methylase METTL3 was sufficient to promote cardiomyocyte hypertrophy both in vitro and in vivo. Finally, cardiac-specific METTL3 knockout mice exhibit morphological and functional signs of heart failure with aging and stress, showing the necessity of RNA methylation for the maintenance of cardiac homeostasis.

Conclusions:

Our study identified METTL3-mediated methylation of mRNA on
N
6
-adenosines as a dynamic modification that is enhanced in response to hypertrophic stimuli and is necessary for a normal hypertrophic response in cardiomyocytes. Enhanced m6A RNA methylation results in compensated cardiac hypertrophy, whereas diminished m6A drives eccentric cardiomyocyte remodeling and dysfunction, highlighting the critical importance of this novel stress-response mechanism in the heart for maintaining normal cardiac function.
Endothelial Hypoxia-Inducible Factor-2α Is Required for the Maintenance of Airway Microvasculature
Authors: Jiang Xinguo , Tian Wen , Tu Allen B. , Pasupneti Shravani , Shuffle Eric , Dahms Petra , Zhang Patrick , Cai Haoliang , Dinh Thanh T. , Liu Bo , Cain Corey , Giaccia Amato J. , Butcher Eugene C. , Simon M. Celeste , Semenza Gregg L. , Nicolls Mark R.
Published: January 2019

Background:

Hypoxia-inducible factors (HIFs), especially HIF-1α and HIF-2α, are key mediators of the adaptive response to hypoxic stress and play essential roles in maintaining lung homeostasis. Human and animal genetics studies confirm that abnormal HIF correlates with pulmonary vascular pathology and chronic lung diseases, but it remains unclear whether endothelial cell HIF production is essential for microvascular health. The large airway has an ideal circulatory bed for evaluating histological changes and physiology in genetically modified rodents.

Methods:

The tracheal microvasculature of mice, with conditionally deleted or overexpressed HIF-1α or HIF-2α, was evaluated for anatomy, perfusion, and permeability. Angiogenic signaling studies assessed vascular changes attributable to dysregulated HIF expression. An orthotopic tracheal transplantation model further evaluated the contribution of individual HIF isoforms in airway endothelial cells.

Results:

The genetic deletion of
Hif-2α
but not
Hif-1α
caused tracheal endothelial cell apoptosis, diminished pericyte coverage, reduced vascular perfusion, defective barrier function, overlying epithelial abnormalities, and subepithelial fibrotic remodeling. HIF-2α promoted microvascular integrity in airways through endothelial angiopoietin-1/TIE2 signaling and Notch activity. In functional tracheal transplants, HIF-2α deficiency in airway donors accelerated graft microvascular loss, whereas HIF-2α or angiopoietin-1 overexpression prolonged transplant microvascular perfusion. Augmented endothelial HIF-2α in transplant donors promoted airway microvascular integrity and diminished alloimmune inflammation.

Conclusions:

Our findings reveal that the constitutive expression of endothelial HIF-2α is required for airway microvascular health.
Association Between Sleep Apnea and Blood Pressure Control Among Blacks
Authors: Johnson Dayna A. , Thomas S. Justin , Abdalla Marwah , Guo Na , Yano Yuichiro , Rueschman Michael , Tanner Rikki M. , Mittleman Murray A. , Calhoun David A. , Wilson James G. , Muntner Paul , Redline Susan
Published: January 2019

Background:

Blacks have a high prevalence of hypertension and uncontrolled blood pressure (BP), each of which may be partially explained by untreated sleep apnea. We investigated the association of sleep apnea with uncontrolled BP and resistant hypertension in blacks.

Methods:

Between 2012 and 2016, Jackson Heart Sleep Study participants (N=913) underwent an in-home Type 3 sleep apnea study, clinic BP measurements, and anthropometry. Moderate or severe obstructive sleep apnea (OSA) was defined as a respiratory event index ≥15, and nocturnal hypoxemia was quantified as percent sleep time with <90% oxyhemoglobin saturation. Prevalent hypertension was defined as either a systolic BP ≥130 mm Hg or diastolic BP
>
80mm Hg, use of antihypertensive medication, or self-report of a diagnosis of hypertension. Controlled BP was defined as systolic BP <130 mm Hg and diastolic BP <80 mm Hg; uncontrolled BP as systolic BP ≥130 mm Hg or diastolic BP ≥80 mm Hg with use of 1 to 2 classes of antihypertensive medication; and resistant BP as systolic BP ≥130 mm Hg or diastolic BP ≥80 mm Hg with the use of ≥3 classes of antihypertensive medication (including a diuretic) or use of ≥4 classes of antihypertensive medication regardless of BP level. Multinomial logistic regression models were fit to determine the association between OSA severity and uncontrolled BP or resistant hypertension (versus controlled BP) after multivariable adjustment.

Results:

The analytic sample with hypertension (N=664) had a mean age of 64.0 (SD,10.6) years, and were predominately female (69.1%), obese (58.6%), and college educated (51.3%). Among the sample, 25.7% had OSA, which was untreated in 94% of participants. Overall, 48% of participants had uncontrolled hypertension and 14% had resistant hypertension. After adjustment for confounders, participants with moderate or severe OSA had a 2.0 times higher odds of resistant hypertension (95% confidence interval [CI], 1.14–3.67). Each standard deviation higher than <90% oxyhemoglobin saturation was associated with an adjusted odds ratio for resistant hypertension of 1.25 (95% CI 1.01–1.55). OSA and <90% oxyhemoglobin saturation were not associated with uncontrolled BP.

Conclusion:

Untreated moderate or severe OSA is associated with increased odds of resistant hypertension. These results suggest that untreated OSA may contribute to inadequate BP control in blacks.