bluuebunny/crossref_metadata_2025 huggingface dataset:
Caveolin-1 expression predicts favourable outcome and correlates withPDGFRAmutations in gastrointestinal stromal tumours (GISTs)
Authors: Bertero Luca , Gambella Alessandro , Barreca Antonella , Osella-Abate Simona , Chiusa Luigi , Francia di Celle Paola , Lista Patrizia , Papotti Mauro , Cassoni Paola
Published: June 2021

Aims

Novel prognostic markers are warranted for gastrointestinal stromal tumours. Caveolin-1 is a multifunctional protein that proved to be associated with outcome in multiple tumour types. Aim of this study was to investigate Caveolin-1 expression and prognostic efficacy in a series of gastrointestinal stromal tumours.

Methods

Caveolin-1 expression was assessed by immunohistochemistry in a retrospective series of 66 gastrointestinal stromal tumours representative of the different molecular subtypes. Correlations with clinical, histopathological and molecular features were investigated. Statistical analyses were performed as appropriate.

Results

Thirty-five cases out of 66 (53.0%) expressed Caveolin-1. Presence of Caveolin-1 expression correlated with favourable histopathologic and clinical traits, including a lower mitotic count (p=0.003) and lower relapse rate (p=0.005). Caveolin-1 expression also resulted associated with the presence ofPDGFRAmutations (p=0.010). Outcome analyses showed a favourable prognostic significance of Caveolin-1 expression in terms of relapse-free survival (HR=0.14; 95% CI=0.03 to 0.63) and overall survival (HR=0.29; 95% CI=0.11 to 0.74), even after adjusting for the mutational subgroup (relapse-free survival: HR=0.14, 95% CI=0.04 to 0.44; overall survival: HR=0.29, 95% CI=0.11 to 0.51) and imatinib treatment (relapse-free survival: HR=0.14, 95% CI=0.02 to 0.81; overall survival: HR=0.29, 95% CI=0.17 to 0.48).

Conclusion

Caveolin-1 represents a novel prognostic marker in gastrointestinal stromal tumours. Further studies are warranted to validate these results and to explore the mechanisms linking Caveolin-1 expression with thePDGFRAoncogenic pathway.
Elastodynamic Analysis of Functionally Graded Beams and Plates Based on Meshless RKPM
Authors: Qin Shaopeng , Wei Gaofeng , Liu Zheng , Shen Xuehui
Published: June 2021

In this paper, the reproducing kernel particle method (RKPM) is innovatively extended to the elastodynamic analysis of functionally graded material (FGM). The elastodynamics governing equations of FGM are solved by using the RKPM. The penalty factor method is used to solve the displacement boundary conditions, and the Newmark-[Formula: see text] method is used to discretize the time. The influence of the penalty factor and the scaling parameter is discussed, and the stability and convergence of the RKPM are analyzed. Finally, the correctness of meshless RKPM to solve the elastodynamics of FGM is verified by numerical examples of the functionally graded beams and plates.
Robust Networking: Dynamic Topology Evolution Learning for Internet of Things
Authors: Chen Ning , Qiu Tie , Daneshmand Mahmoud , Wu Dapeng Oliver
Published: June 2021

The Internet of Things (IoT) has been extensively deployed in smart cities. However, with the expanding scale of networking, the failure of some nodes in the network severely affects the communication capacity of IoT applications. Therefore, researchers pay attention to improving communication capacity caused by network failures for applications that require high quality of services (QoS). Furthermore, the robustness of network topology is an important metric to measure the network communication capacity and the ability to resist the cyber-attacks induced by some failed nodes. While some algorithms have been proposed to enhance the robustness of IoT topologies, they are characterized by large computation overhead, and lacking a lightweight topology optimization model. To address this problem, we first propose a novel robustness optimization using evolution learning (ROEL) with a neural network. ROEL dynamically optimizes the IoT topology and intelligently prospects the robust degree in the process of evolutionary optimization. The experimental results demonstrate that ROEL can represent the evolutionary process of IoT topologies, and the prediction accuracy of network robustness is satisfactory with a small error ratio. Our algorithm has a better tolerance capacity in terms of resistance to random attacks and malicious attacks compared with other algorithms.
Promoters and Antagonists of Phagocytosis: A Plastic and Tunable Response
Authors: Freeman Spencer , Grinstein Sergio
Published: June 2021

Recent observations indicate that, rather than being an all-or-none response, phagocytosis is finely tuned by a host of developmental and environmental factors. The expression of key phagocytic determinants is regulated via transcriptional and epigenetic means that confer memory on the process. Membrane traffic, the cytoskeleton, and inside-out signaling control the activation of phagocytic receptors and their ability to access their targets. An exquisite extra layer of complexity is introduced by the coexistence of distinct “eat-me” and “don't-eat-me” signals on targets and of corresponding “eat” and “don't-eat” receptors on the phagocyte surface. Moreover, assorted physical barriers constitute “don't-come-close-to-me” hurdles that obstruct the engagement of ligands by receptors. The expression, mobility, and accessibility of all these determinants can be modulated, conferring extreme plasticity on phagocytosis and providing attractive targets for therapeutic intervention in cancer, atherosclerosis, and dementia.
Three weeks of interrupting sitting lowers fasting glucose and glycemic variability, but not glucose tolerance, in free-living women and men with obesity
Authors: Smith Jonathon A. B. , Savikj Mladen , Sethi Parneet , Platt Simon , Gabriel Brendan M. , Hawley John A. , Dunstan David , Krook Anna , Zierath Juleen R. , Näslund Erik
Published: June 2021

Under free-living conditions, breaking sitting modestly increased activity behavior. Breaking sitting was insufficient to modulate glucose tolerance or the skeletal muscle lipidome. Activity breaks reduced fasting blood glucose levels and daily glucose variation compared with baseline, with a tendency to also decrease fasting LDLc. This intervention may represent the minimal dose for breaking sedentary behavior, with larger volumes of activity possibly required to promote greater health benefits.
KIM-1-mediated anti-inflammatory activity is preserved by MUC1 induction in the proximal tubule during ischemia-reperfusion injury
Authors: Al-bataineh Mohammad M. , Kinlough Carol L. , Mi Zaichuan , Jackson Edwin K. , Mutchler Stephanie M. , Emlet David R. , Kellum John A. , Hughey Rebecca P.
Published: June 2021

KIM-1 plays a key role in the recovery of the tubule epithelium during renal IRI by mediating efferocytosis and associated signaling that suppresses inflammation. Excessive cleavage of KIM-1 by ADAM17 provides a decoy receptor that aggravates efferocytosis and subsequent signaling. Our data from experiments in mice, patients, and cultured cells show that MUC1 is also induced during IRI and competes with KIM-1 for cleavage by ADAM17. Consequently, MUC1 protects KIM-1 anti-inflammatory activity in the damaged kidney.
The Thermal Performance of Chinese Vernacular Skywell Dwellings
Authors: Yao Gang , Han Dongting , Zhang Li , Duan Zhongcheng
Published: June 2021

The aim of this research was to investigate the thermal performance of Chinese vernacular skywell dwellings. This was done by the analysis of on‐site measurements of the environmental performance and computer simulation of solar conditions of eight vernacular skywell dwellings in three villages in southeastern China—Xidi, Zhifeng, and Yuyuan. Through the discussion of several heat inputs and outputs, it can be concluded that evaporative cooling had a substantial influence on the temperature in the skywells of dwellings in Xidi and Zhifeng villages. In addition to the buffering of temperature by thermal mass, evaporative cooling was found to further reduce the fluctuation in temperature inside the skywell and is likely to have been the main reason that the mean dry bulb temperatures inside the skywells in these villages were lower than the mean external dry bulb temperatures. Moreover, the construction materials of Chinese vernacular dwellings (skywell wall, floor material, wooden column, and panels) are likely to be hygroscopic.
Evolution of Strength Parameters for Sandstone Specimens during Triaxial Compression Tests
Authors: Ding Xiang , Chen Na , Zhang Fan , Zhang Guangqing
Published: June 2021

Despite the lack of test data of the coefficient of pressure sensitivity α and the shearing cohesion k, the Drucker–Prager criterion is commonly applied for numerical analyses of geotechnical engineering. To bridge the gap between the wide application and insufficient knowledge of strength parameters of the Drucker–Prager criterion, this study presents experimentally calibrated strength parameters of this criterion for the first time. This paper proposes a new method to measure α and k in the Drucker–Prager criterion. The square root of the second invariant of the deviatoric stress tensor is linearly fitted with the first invariant of the stress tensor I1 in the stress space. The parameters φ and c in the Mohr–Coulomb criterion and α and k in the Drucker–Prager criterion are calibrated to the same set of triaxial compression tests of sandstones. With these testing results, five pairs of conversion formulae (which are most commonly used in the literature) are examined and the most appropriate pair of conversion formulae is identified. With parameters indicating cohesive strength (c and k) and parameters indicating frictional strength (φ and α), the evolutions of different strength components are compared with those in the cohesion‐weakening friction‐strengthening model. With an increase in plastic deformation, the cohesive strength parameters c and k firstly increase to a peak value and then decrease. The frictional strength parameters φ and α gradually increase at a decreasing rate after the initial yield point.
Numerical Investigation into the Distributions of Temperature and Stress around Wellbore during the Injection of Cryogenic Liquid Nitrogen into Hot Dry Rock Reservoir
Authors: Ren Keda , Cai Chengzheng
Published: June 2021

Cryogenic liquid nitrogen fracturing is expected to provide an effective stimulation method for hot dry rock reservoirs to increase heat production. This paper establishes a three-dimensional model to calculate the distributions of temperature and stress of the reservoir rock when liquid nitrogen is injected into the wellbore. The sensitivity of different parameters and water fracturing to the stress state is studied. The results indicate that when liquid nitrogen is injected into the bottom of well, a huge heat exchange occurs on the rock surface, which generates great thermal stress on the fluid-solid interface, and the value of thermal stress exceeds the tensile strength of rock. For the effect of parameters, the primitive temperature of the rock has a significant impact on the value of maximum principal stress. The pressure drop and ambient pressure affect the thermal stress slightly. At the same time, a series of experiments are conducted to validate the effect of thermal stress induced by liquid nitrogen injection on the rock fracture. As the temperature rises, the shale samples are broken more severely at the action of thermal stress. Thus, the study of liquid nitrogen fracturing provides a scientific and effective method for geothermal exploitation.
Biotransformation of Daidzein, Genistein, and Naringenin by Streptomyces Species Isolated from High-Altitude Soil of Nepal
Authors: Shrestha Lasata , Marasini Bishnu P. , Pradhan Suman Prakash , Shrestha Rajib Kumar , Shrestha Suraj , Regmi Kamal Prasad , Pandey Bishnu Prasad
Published: June 2021

Flavonoids have achieved widespread importance in pharmaceutical, food, and cosmetics industries. Furthermore, modification of these naturally occurring flavonoids to structurally diverse compounds through whole cell biotransformation with enhanced biological activities has numerous biotechnological applications. The present study investigated the biotransformation potential of Streptomyces species isolated from a high-altitude-soil sample towards selected flavonoid molecules. The biotransformed metabolites were confirmed by comparing the HPLC chromatogram with authentic compounds and LC-MS/MS analysis. Of these isolates, Streptomyces species G-18 (Accession number: MW663767.1) catalyzed isoflavone molecules daidzein and genistein to produce hydroxylated products at 24 h of reaction condition in a whole cell system. The hydroxylation of daidzein (4′,7-dihydroxyisoflavone) was confirmed at 3′-position of the B ring to produce 3′,4′,7-trihydroxyisoflavone. In addition, Streptomyces species G-14 (Accession number: MW663770.1) and Streptomyces species S4L (Accession number: MW663769.1) also revealed the transformation of daidzein (4′,7-dihydroxyisoflavone) to hydroxy daidzein at a distinct position than that of G-18 isolates, whereas thee Streptomyces species S4L reaction mixture with naringenin as a substrate also revealed the hydroxylated product. Our results demonstrated that microorganisms isolated from different ecological niches have broad application.
Castration-Induced Downregulation of SPARC in Stromal Cells Drives Neuroendocrine Differentiation of Prostate Cancer
Authors: Enriquez Claudia , Cancila Valeria , Ferri Renata , Sulsenti Roberta , Fischetti Irene , Milani Matteo , Ostano Paola , Gregnanin Ilaria , Mello-Grand Maurizia , Berrino Enrico , Bregni Marco , Renne Giuseppe , Tripodo Claudio , Colombo Mario P. , Jachetti Elena
Published: June 2021

Fatal neuroendocrine differentiation (NED) of castration-resistant prostate cancer is a recurrent mechanism of resistance to androgen deprivation therapies (ADT) and antiandrogen receptor pathway inhibitors (ARPI) in patients. The design of effective therapies for neuroendocrine prostate cancer (NEPC) is complicated by limited knowledge of the molecular mechanisms governing NED. The paucity of acquired genomic alterations and the deregulation of epigenetic and transcription factors suggest a potential contribution from the microenvironment. In this context, whether ADT/ARPI induces stromal cells to release NED-promoting molecules and the underlying molecular networks are unestablished. Here, we utilized transgenic and transplantable mouse models and coculture experiments to unveil a novel tumor-stroma cross-talk that is able to induce NED under the pressure of androgen deprivation. Castration induced upregulation of GRP78 in tumor cells, which triggers miR29-b–mediated downregulation of the matricellular protein SPARC in the nearby stroma. SPARC downregulation enabled stromal cells to release IL6, a known inducer of NED. A drug that targets GRP78 blocked NED in castrated mice. A public, human NEPC gene expression dataset showed that Hspa5 (encoding for GRP78) positively correlates with hallmarks of NED. Finally, prostate cancer specimens from patients developing local NED after ADT showed GRP78 upregulation in tumor cells and SPARC downregulation in the stroma. These results point to GRP78 as a potential therapeutic target and to SPARC downregulation in stromal cells as a potential early biomarker of tumors undergoing NED.

Significance:

Tumor–stroma cross-talk promotes neuroendocrine differentiation in prostate cancer in response to hormone therapy via a GRP78/SPARC/IL6 axis, providing potential therapeutic targets and biomarkers for neuroendocrine prostate cancer.
Knowledge of Diabetes mellitus and Glycaemic Control
Authors: Robinson David L. , Al-Bustan Mahmoud , Bitar Milad S. , Al-Asousi Adnan , Majeed Sobia
Published: June 2021

It has been claimed that lack of knowledge of diabetes mellitus has been a cause of excess admissions and morbidity. There is also some evidence that diabetic education programs can improve self-regulatory behaviour and reduce hospital admissions. In this report we test the hypothesis that greater knowledge of diabetes should be associated with better glycaemic control and lower fasting blood glucose levels. Responses to diabetes knowledge questions were provided by 420 patients attending diabetic clinics in Kuwait. In an earlier and complementary report a principal components analysis revealed that knowledge of diabetes cannot be understood in terms of a single general factor. With a subsequent Varimax rotation we obtained 12 uncorrelated knowledge factors with eigenvalues greater than unity and these would all be confounded in the simple aggregation of correct answers to diabetes knowledge questions employed in earlier studies. Results are now described which show that the 12-factor model of diabetes knowledge is better able to predict blood glucose levels than the scores obtained on a single scale by just summing the correct answers to all diabetes knowledge questions. A standard multiple linear regression with the diabetes knowledge factors age, sex and ‘years since diagnosis’ as independent variables, and fasting blood glucose levels as the dependent variable, shows that 4 of the 12 factors yield statistically significant semi-partial correlation coefficients that account for unique fractions of the total variance of blood glucose levels. The meaning of these 4 factors is discussed with special reference to glycaemic control and blood glucose levels.
Treatment of Optic Canal Decompression Combined with Umbilical Cord Mesenchymal Stem (Stromal) Cells for Indirect Traumatic Optic Neuropathy: A Phase 1 Clinical Trial
Authors: Li Jia , Bai Xu , Guan Xiaoyue , Yuan Hongfeng , Xu Xiang
Published: June 2021

Purpose: This study was aimed to investigate the safety and feasibility of umbilical cord-derived mesenchymal stem cell (MSC) transplantation in patients with traumatic optic neuropathy (TON). Methods: This is a single-center, prospective, open-labeled phase 1 study that enrolled 20 patients with TON. Patients consecutively underwent either optic canal decompression combined with MSC local implantation treatment (group 1) or only optic canal decompression (group 2). Patients were evaluated on the first day, seventh day, first month, third month, and sixth month postoperatively. Adverse events, such as fever, urticarial lesions, nasal infection, and death, were recorded at each visit. The primary outcome was changes in best-corrected visual acuity. The secondary outcomes were changes in color vision, relative afferent pupillary defect, and flash visual evoked potential. Results: All 20 patients completed the 6-month follow-up. None of them had any systemic or ocular complications. The change in best-corrected visual acuity at follow-up was not significantly different between group 1 and group 2 (p > 0.05); however, group 1 showed better visual outcome than group 2. Both groups showed significant improvements in vision compared with the baseline (p < 0.05); however, there were no statistically significant differences between the groups (p > 0.05). In addition, no adverse events related to local transplantation were observed in the patients. Conclusions: A single, local MSC transplantation in the optic nerve is safe for patients with TON.
Indagações acerca dos marcadores culturais na tradução
Authors: Aubert Francis Henrik
Published: June 2021

Nos estudos descritivos da tradução, a identificação dos marcadores culturais depara-se com certas dificuldades teóricas e metodológicas: a pró- pria conceituação de marcador cultural; suas subcategorias, lingüísticas e ex- tralingüísticas; os procedimentos apropriados para proceder à sua identifica- ção. O presente ensaio procura mapear a extensão dessas dificuldades e pro- por alguns encaminhamentos, ainda a serem testados na prática descritiva.
Abstract 102: Number of Reentry Tears Influences Flap Motion and Flow Reversal in an
in vitro
Model of Type B Aortic Dissection
Authors: Birjiniuk Joav , Young Mark , Timmins Lucas H , Leshnower Bradley G , Oshinski John N , Ku David N , Veeraswamy Ravi K
Published: June 2021

Objectives:
Aortic remodeling after dissection is poorly understood. Thus, optimal patient-specific recommendations for treatment are lacking. An
in vitro
aortic model of Type B dissection was used to interrogate local aortic hemodynamic parameters implicated in thrombosis and aneurysm formation. We hypothesize that dissections with multiple reentry tears will exhibit decreased flap motion, and, as a result, reduce flow reversal.

Methods:
Anatomic models of aortic dissection with fidelity to patient CT images were fabricated out of silicone. Models with primary entry and single fenestration (Figure 1A), two fenestrations (Figure 1B), and three fenestrations (Figure 1C) were installed in a flow loop. Physiologic flow was established at a cardiac index of 4 L/min. Flow velocities were acquired using phase contrast magnetic resonance (PCMR) imaging. Flow rates and flap motion were quantified using custom made software.

Results:
Relative true lumen area (RTLA) varied along the dissection (entry: 55% +/- 3, middle: 34% +/-7, exit: 91%+/-3, p<0.00001 pair-wise for 2-tear model). At mid-dissection, RTLA was lower in dissections with fewer tears (p<0.01). Total flow was nearly identical in all cases, while true and false lumen flow rates differed significantly across tear configurations and along the dissection (p<0.01). Secondary tears allowed for flow communication within the dissected portion of the aortic model. Flow reversal was seen in the false lumen at the mid-dissection plane in the absence of secondary tears (Figure 1D). However, as secondary tears were added, the flow reversal in the false lumen decreased, with concomitant flow reversal in the true lumen (Figure 1E,F).

Conclusions:
Anatomic characteristics of dissection, such as number of tears, affect blood flow and motion of the dissection flap, as shown quantitatively. This compliant aorta model illustrates alterations in flow reversal in both true and false lumina that may lead to aneurysmal degeneration.
Abstract 112: Low Plasma Low-density Lipoprotein Cholesterol Level as a Risk Factor for Cancer in Patients With Newly Diagnosed Coronary Artery Disease
Authors: Chen Xiaofeng , Jiang Jianjun , Yang Haihua , Wang Bing , Xue Yinsheng , Liu Xiaowei , Tang Lijiang
Published: June 2021

Background and Objective:
Coronary artery disease (CAD) and cancer are the leading causes of death worldwide and share some risk factors, such as aging and smoking. Concurrence of CAD and cancers makes diagnosis and treatment more clinical challenging. In patients who need stent implantation followed by dual antiplatelet therapy, fatal perioperative complications such as bleeding and stent thrombosis may occur. There is no data regarding the prevalence and risk factors of cancer in patients with CAD. This study investigated the cancer prevalence and risk factors in patients with newly diagnosed CAD.

Methods and Results:
Three thousand and one hundred and eighty six consecutive patients with newly angiographically documented CAD were prospectively enrolled between January 2009 and March 2015. Serum levels of tumor markers, including carcinoembryonic antigen, alpha fetoprotein, carbohydrate antigen125, carbohydrate antigen 153, carbohydrate antigen199, squamous cell cancer antigen, and prostate-specific antigen, were measured. Diagnosis of cancer was confirmed by pathology or imaging with ultrasound, computed tomography or magnetic resonance. The prevalence of cancer in CAD patients was 1.1% (35 out of 3186), involving the in gastroenterogical, respiratory, endocrine and exocrine, urinary and hematopoietic systems. Among these cancer patients, 17 patients had stent implantation, with 7 AMI patients undergoing emergency PCI. Multivariate analysis revealed that, in addition to male gender and advanced age, lower plasma low-density lipoprotein (LDL) cholesterol levels were independently associated with the occurrence of cancer in CAD patients (P<0.01).

Conclusion:
Cancers occur in 1.1% of patients with CAD. Low LDL levels may increase the risk for cancers in the CAD patients. Further large scale studies are needed to validate this association.
Abstract 12: Nanotechnology-enabled Anti-mir-320 Therapy for Inhibiting Pathological Vasoconstriction
Authors: Werfel Thomas A , Hocking Kyle M , Kilchrist Kameron V , Brophy Colleen , Duvall Craig L
Published: June 2021

Subarachnoid hemorrhage (SAH) affects approximately 30,000 people each year and accounts for 1-7% of all strokes. Current SAH therapies globally affect blood pressure, potentially causing hypotension and reduced cranial perfusion. Alternatively, our approach is to locally reduce constriction of the blood vessels in the brain by targeting heat shock protein 20 (HSP 20) which plays a role in the thin filament pathway for contraction and relaxation. To this end, we developed nanoparticles for delivery of a locked nucleic acid (LNA, LNA-NPs) designed to inhibit miR-320, an endogenous negative regulator of HSP 20 expression in vascular tissue. We hypothesized that LNA-NP-enabled anti-miR-320 therapy would increase HSP 20 expression, reducing vessel contraction and increasing vessel relaxation. LNA-NPs were formulated with a PEGylated, endosomolytic polymer which has been previously optimized for systemic siRNA delivery (Fig. 1A). In this study, LNA-NPs efficiently complexed LNA (Fig. 1B) and formed ~100 nm diameter LNA-loaded nanoparticles (Fig. 1C). Efficient cytosolic delivery of LNA by LNA-NPs resulted in ~2.5-fold (n = 3, Fig. 1D) and 21% (n = 4, Fig. 1E) increased HSP 20 protein expression compared to LNA alone in rat aortic smooth muscle cells and rat aortic vessels, respectively. Functionally, LNA-NP treated tissue showed a 21% reduction in phenylephrine induced contraction relative to maximal 110 mM KCl-induced contraction (n = 4, Fig. 1F). Rat aorta treated with LNA-NPs also showed increased relaxation within the vessels to sodium nitroprusside by 8.1% (n=4, Fig. 1F). In sum, this approach allowed inhibition of miR-320 and direct modulation of HSP 20 in a translationally relevant organ culture model of vascular disease. Future studies will focus on inhibiting miR-320 in human saphenous vein
ex vivo
and in a pre-clinical model of SAH after systemic delivery.
Abstract 129: Overexpression of P110alpha Induces Vascular Malformation in Mouse Tail, Feet and Ears
Authors: Babaev Vladimir R , Brooksbank Jeremy A , Ding Lei , Zhang Youmin , May James M , Linton MacRae F
Published: June 2021

Phosphoinositide-3 kinases (PI3Ks) are a family of signal transduction enzymes involved in critical cellular functions such as cell growth, proliferation, differentiation, motility and survival. Class IA PI3K is composed of a heterodimer between a P110 catalytic subunit and a p85 regulatory subunit. Upregulation of P110alpha protein significantly propagates second messenger response accelerating Akt signaling. To augment Akt activity in macrophages, we generated mice with conditional myeloid lineage-specific (LysM-Cre) overexpression of P110alpha (M-P110α+). Peritoneal macrophages isolated from these mice expressed an additional band slightly different in size than P110alpha protein and had markedly increased phosphorylation of Akt. These mice also exhibited dramatic changes in blood cells including lower levels of white blood cells, B-cells and an increase in neutrophils; whereas the levels of T-cells and monocytes were not different compared to wild-type mice. Because of the defects of B-cell development, M-P110α+ mice had also splenomegaly with decreased levels of follicles. Unexpectedly, M-P110α+ mice at age of 3-4 weeks spontaneously developed a striking phenotype with massive enlargements of small vessels forming malformations in the extremities including tail, feet and ears. These vascular malformations grew very rapidly, and, at age 5-7 months, M-P110α+ mice developed severe hemorrhages from lesions on the tail and feet. The malformations appeared to be mainly venous by Doppler color ultrasonography, and histologically they represented overgrowth of venous and capillary cells that expressed endothelial markers including CD31/PECAM-1, von Willibrand factor and panendothelial cell antigen, MECA-32. These endothelial cells exhibited high levels of proliferation detected by Ki67 staining. In addition, the malformations were surrounded by high numbers of macrophages. In conclusion, M-P110α+ mice develop spontaneous capillary-venous malformations that represent a novel model of the human PIK3CA-related overgrowth spectrum. This mouse model provides a unique opportunity to develop novel therapeutic strategies to prevent capillary and venous malformations.
Abstract 13: Targeting the PERK Pathway of ER Stress Response for Endothelium Protection and Restenosis Prevention:
A Paradigm for Developing Anti-thrombogenic Stents
Authors: Wang Bowen , Zhang Mengxue , Shi Xudong , Guo Lian-Wang , Hoffmann Michael , Kent Craig
Published: June 2021

Introduction:
Cardiovascular disease is the leading cause of mortality and morbidity in the US. Reconstructions often fail due to the recurrent lumen narrowing, or restenosis. Stents eluting rapamycin or paclitaxel are deployed to inhibit the excessive proliferation of smooth muscle cells (SMCs), which is the core component of restenosis. However, these drugs cause collateral damage to endothelial cells (ECs) leading to stent thrombosis. To address this, our group initiated the first campaign of drug screening for compounds that selectively inhibit SMC proliferation without causing EC dysfunction. We recently identified lead compounds as such, which are inhibitors for protein kinase RNA-like endoplasmic reticulum kinase (PERK), an endoplasmic reticulum (ER) stress sensor. Here we evaluated PERK as a target for intervention of both SMC pathophysiology and EC dysfunction.

Methods:
Rat carotid artery balloon angioplasty was performed as a restenosis model. PDGF-BB and TNF-a were applied to primary human aortic SMCs and ECs, respectively, to mimic the
in vivo
pathogenic stimuli.

Results:
In balloon-injured arteries, both PERK phosphorylation and the expression of its downstream transcription factor ATF4 were increased compared to sham control. PERK activation was also observed
in vitro
in both SMCs and ECs stimulated with PDGF-BB and TNF-a, respectively. In SMCs, either selective inhibition (1μM GSK2606414) or siRNA knockdown of PERK abolished PDGF-BB induced de-differentiation and proliferation. In ECs, PERK antagonism abrogated TNF-a induced growth impairment and EC secretion of Tissue Factor (TF), which is the key initiator of thrombogenesis. Finally, in a pilot experiment, peri-adventitial application of GSK2606414 (25mg/kg, n=2 rats) reduced intima/media ratio by 80% and increased lumen area by ~ 2 fold compared to vehicle control (n=2) at 4 weeks post injury.

Conclusion:
Our results indicate an important role of PERK activation in promoting SMC phenotype switching and EC dysfunction in
vitro
as well as restenosis
in vivo
. Thus, PERK targeting represents a potential strategy to simultaneously achieve restenosis prevention and endothelium protection, with a long-term goal of developing anti-thrombogenic drug-eluting stents.
Abstract 133: Mechanistic Insights Into Bradykinin and Thromboxane Receptors Heterodimerization in Vascular Smooth Muscle Cells
Authors: Dagher Oula K , Jaffa Miran A , Habib Aïda , Ziyadeh Fuad N , Luttrell Louis M , Jaffa Ayad A
Published: June 2021

The development of an atherosclerotic lesion in an injured vasculature is highly dependent on the proliferative state of vascular smooth muscle cells (VSMC). Bradykinin (BK) and Thromboxane are two G-protein coupled receptor ligands, whose individual binding to their respective receptors, B
2
R and TP, promotes ERK1/2-mediated VSMC proliferation. However, it is not yet known whether receptor-receptor interactions between B
2
R and TP could contribute to their co-regulation. Thus, our work addresses the hypothesis that, in VSMC, B
2
R and TP form functional hetero-complexes, of distinct trafficking and signaling properties. B
2
R-TP signaling crosstalk was first analyzed in rat VSMC by Western Blot analysis and subsequent fold/basal quantification of ERK1/2 phosphorylation. B
2
R-TP cooperation was evident through the synergistic ERK1/2 phosphorylation, in VSMC co-stimulated with optimized combinations of BK and the TP agonist, IBOP (21.05 ± 4.93 fold of basal, n=3; p < 0.001). Interestingly, however, pretreatment with the TP antagonist, SQ29548, totally abolished (BK+IBOP)-induced ERK1/2 (n=3; p < 0.001). Furthermore, knowing that B
2
R, unlike the human TPα isoform, undergoes agonist-induced sequestration and β-arrestin2 recruitment, we conducted immunofluorescence analysis on HEK293T cells overexpressing human B
2
R and human TPα (B
2
R-TPα-HEKs). While stimulation with IBOP failed to mobilize cell membranous B
2
R or TPα, substantial co-internalization of TPα and B
2
R was seen, subsequent to stimulation of B
2
R-TPα-HEKs with BK. Likewise, overexpressing β-arrestin2 in B
2
R-TPα-HEKs resulted in co-localization of B
2
R, TPα, and β-arrestin2 within internalized puncta, only after BK stimulation. Pretreatment with SQ29548 inhibited BK-induced co-internalization and β-arrestin2 recruitment. However, SQ29548 could not reverse BK-induced B
2
R sequestration in HEK293T cells overexpressing B
2
R alone, thus excluding the possibility of direct SQ29548 - B
2
R binding. Finally, results of our ongoing work are the first to show nuclear localization of B
2
R and TP within human and rat VSMC. Thus, our findings favor the likelihood of functional B
2
R-TP heterodimerization in VSMC, which could serve as a novel target for interventional strategies.
Abstract 14: The Anti-atherosclerosis ABCA1 Agonist CS6253 Confer Glucose Control by Improved Pancreas Beta-cell Insulin Secretion and Enhanced Peripheral Insulin Utility
Authors: Bielicki John K , Hafiane Anouar , Genest Jacques , Johansson Jan O , Bittner Stefanie , Hu Jie , Tabassum Juveri , Azhar Salman
Published: June 2021

Background:
Type 2 Diabetes Mellitus (T2DM) is associated with high cardiovascular disease (CVD) risk. Addressing the underlying atherogenesis and diabetes causing CVD in T2DM is important. CS6253 is an ABCA1 agonist peptide derived from the C-terminal of apoE that has shown macrophage specific reverse cholesterol transport, anti-atherosclerosis and anti-diabetic properties. Further studies were carried out to characterize metabolic effects.

Methods:
CS6253 was incubated with a) INS-1 823/13 cells to assess effects on insulin secretion and b) with L6-Glut4myc rat myoblasts to assess glucose uptake properties. Diet Induced Obesity (DIO) mice, i.e. C57BL/6 mice that had been fed 60% high-fat diet for 6 weeks, were treated with CS6253 and Glucose Tolerance Tests (GTT) performed after overnights fasting administering glucose 1g/kg ip.

Results:
CS6253 1mg/mL incubated for 2 hours under standard conditions with 3mM glucose showed a 3-fold increase in insulin secretion compared to control, i.e. 232(32) vs. 79(7) ng/M cells, p<0.001.
3
H-glucose uptake by CS6253 peptide in L6-Glut4myc rat myoblasts increased insulin’s glucose uptake capacity from 3800 to 4619 DPM/well, p<0.001 . CS6253 alone had no effect on
3
H-glucose uptake compared to control.

DIO mice were treated with CS6253 30mg/kg sc alternate days or PBS control for 16 weeks. GTTs were performed after 2, 6 and 15 weeks treatment showing 39%, 45% and 57% reductions in the glucose-AUC compared to control, respectively, p<0.01 for all time points. Insulin response to GTT after 5 weeks treatment showed a strong improvement of the insulin-curve by CS6253, p<0.05 vs. placebo. CS6253 treated DIO mice showed a non-significant body weight decrease and a 17% reduction in liver weight, 5.28g vs. 4.36g, p<0.01.

Discussion:
CS6253 shows potent, sustained and increased anti-diabetic actions over the 16 weeks treatment period in DIO mice. In vivo and in vitro studies show improved pancreas β-cell function with increased glucose-mediated insulin secretion and also insulin sensitizing properties. CS6253’s combined anti-diabetic and anti-atherosclerosis properties suggest utility in the treatment of CVD and T2DM.
Abstract 140: Magnesium Lithospermate B Inhibits Blood Coagulation and Platelet Aggregation: Novel Mechanism of a Traditional Drug for Cardiovascular Diseases
Authors: Wang Yiming , Zhang Dan , Xu Xiaohong , Lei Xi , Wang Yiping , Xuan Lijiang , Ding Jian , Ni Heyu
Published: June 2021

Magnesium lithospermate B (MLB) is one of the major components of Salvia miltiorrhiza root (Danshen). Danshen extracts have been used to control cardiovascular disease for centuries. In 2005, intravenous injection of Danshen depside salt was approved in China for treatment of chronic angina. Although clinical observations have suggested that Danshen extracts inhibited thrombosis, the exact mechanism has not been adequately explored.

Using an
in vitro
whole blood clotting assay, we observed that MLB (250 μM) significantly reduced clot size. Both the clot wet and dry weights were decreased following treatment (108.3 mg
vs.
63.5 mg, and 32.8 mg
vs.
18.5 mg, p<0.05, respectively). Using thromboelastography, we found that MLB markedly decreased the mechanical strength of the clot and modestly delayed initiation of coagulation in cell-free blood plasma prepared by centrifugation (10,000 хg, 10 min). Under confocal microscopy, we further observed that MLB significantly reduced the density of the fibrin network formed in plasma following thrombin treatment, suggesting that MLB targets coagulation factors to inhibit coagulation.

Recent network pharmacology analyses predict that MLB may interact with VWF, factor XIII (FXIII), or thrombin in the coagulation cascade. We found that MLB did not inhibit VWF-dependent platelet agglutination induced by botrocetin. ELISA revealed that MLB also did not significantly alter the binding of activated FXIII (FXIIIa) to fibrinogen. However, when native FXIII from blood plasma was used for the same assay, MLB significantly reduced the binding of FXIIIa to fibrinogen. Since generation of FXIIIa from FXIII is thrombin-dependent, these data suggest that MLB inhibits thrombin activity or thrombin generation. Indeed, we found that MLB markedly inhibited thrombin-induced gel-filtered human and mouse platelet aggregation.

These data demonstrated a novel role for MLB in the inhibition of blood coagulation and platelet aggregation, likely through direct inhibition of thrombin function, although we cannot exclude its additional anti-thrombotic activities. Thus, purified MLB may represent an efficient, low-cost agent for treatment of artery and deep vein thrombosis.
Abstract 144: A Single Nucleotide Polymorphism of p27
Kip1
Associated with Vein Graft Patency Regulates Expression of p27
Kip1
in Both Venous Adventitial Cells and Smooth Muscle Cells but Selectively Regulates Proliferation of Adventitial Cells
Authors: Kenagy Richard D , Kikuchi Shinsuke , Chen Lihua , Wijelath Errol S , Clowes Alexander W , Sobel Michael
Published: June 2021

Introduction:
p27
Kip1
(p27) is a cell-cycle inhibitor whose -838C>A single nucleotide polymorphism (SNP) accounts for ~40% of the risk of peripheral vein graft failure. However, whether this SNP is functional has not been definitely established.

Methods:
To determine functionality, we investigated paired adventitial cells and smooth muscle cells (SMC) derived from fresh human saphenous veins (N≥7 lines per group). After growth arrest in 2% serum followed by stimulation with 10 ng/ml PDGF-BB, we measured p27 mRNA, p27 protein, and cell proliferation using qRT-PCR, Western blotting, and cell counts, respectively.

Results:
The SNP genotype was associated with 72 hour adventitial cell growth, but not SMC growth. Adventitial AA cells grew 33% slower than those with the CC genotype (Figure 1A, B, p=.004). We expected AA adventitial cells to produce more p27 mRNA, but paradoxically, p27 mRNA was lower in both cell types of the AA genotype (Figure 1C, D, P<.01). However, levels of p27 protein (a single ~27 kD band in both cell types) were ~2 fold higher in AA adventitial cells and SMCs compared to the CC cells at both 0 and 72 hours (Figure 1E, 72 hours; P<.001 for genotype; 0 h data not shown). Adventitial cells and SMCs made comparable amounts of p27 protein.

Conclusion:
The p27
Kip1
-838C>A SNP regulates levels of p27 in both venous adventitial cells and SMCs. There is an effect of genotype on the growth of adventitial cells, but not on SMCs. AA cells produce ~2 fold higher p27 protein than CC cells, despite AA cells having ~2 fold lower levels of p27 mRNA. These data demonstrate that the p27 SNP is functional, that different cell types within the vein wall respond uniquely to this genetic polymorphism, and suggest an important role in graft failure for the precursor of the cultured adventitial cell.
Abstract 145: Microcapsule Delivery of Mesenchymal Stem Cell for Myocardial Regeneration
Authors: Saraf Anita , Headen Devon , Liu Allen , Weiss Daiana , Brown Milton , Joseph Gigi , Davis Michael , García Andrés J , Taylor W R
Published: June 2021

Following myocardial injury, 1 in 3 patients will have irreversible cardiac damage and develop congestive heart failure. The paracrine factors secreted by mesenchymal stem cells (MSCs) have shown promise in regenerating damaged heart tissue through their pro-angiogenic and anti-inflammatory properties. The beneficial effect of MSCs can be further optimized by their targeted delivery and increased retention within the damaged myocardium.

Objective:
To develop a novel cell therapy approach which allows direct infusion of encapsulated mesenchymal stem cells into the coronary circulation without compromising cardiac function.

Methods:
Mesenchymal stem cells were encapsulated into polyethylene glycol (PEG) based microparticles (MPs) using microfluidics technology. These encapsulated MSCs were infused ex vivo into the rat coronaries using retrograde perfusion in a Langendorff - isolated heart setup. Rat hearts infused with fluorescently tagged MPs were imaged using Xtreme Optical imaging (Bruker, MA) to evaluate for MP distribution within the tissue. Frozen sections of the MP infused hearts were evaluated for distribution within the coronary microvasculature. To evaluate cardiac function after iterative MP infusion, pressure volume curves were generated using a Millar catheter and recorded on Powerlab (ADI instruments, Australia).

Results:
The microfluidics encapsulation process generates microparticles of 30-60 μm diameter with up to 59.7% encapsulation efficiency. Encapsulated MSCs distribute evenly throughout the heart as assessed by whole-organ fluorescence imaging. Systolic function of the rat heart was not compromised when up to 100,000 encapsulated MSCs were infused by retrograde coronary perfusion. Histological sections post infusion showed that the MPs localized to terminal blood vessels as small as 10 μm in diameter, suggesting that these microparticles are compressible and are trapped within small coronary vessels, thereby improving retention.

Conclusion:
Intra-coronary perfusion of encapsulated MSCs is a viable and promising approach for the delivery of MSCs after myocardial injury.
Abstract 15: Identification of a High Density Lipoprotein Proteomic Signature Associated With Atherosclerosis Severity in Humans
Authors: Gordon Scott M , Cui Tiange , Sviridov Denis , Baranova Ancha , Chen Marcus , Remaley Alan T
Published: June 2021

High density lipoprotein associated cholesterol (HDL-C) has been the traditional biomarker for estimating cardiovascular protection derived from HDL; however, several recent studies have made it clear that HDL-C may not necessarily represent the protective capacity of HDL. The diverse protein and lipid content of HDL is becoming increasingly recognized as the basis for the functional complexity observed in these particles. Proteomics studies have identified 95 consensus proteins consistently found to associate with HDL. For the majority of these proteins, the functional importance of their association with HDL has not yet been determined. In this study, we tested the hypothesis that the protein composition of HDL will have an association with atherosclerosis burden in human subjects. A total of 101 subjects were selected based on atherosclerosis severity as determined by CT-angiography: normal (0% stenosis; n = 31), mild (< 30% stenosis; n = 31), moderate (30 – 49% stenosis; n = 30) and severe (>70% stenosis; n = 9). HDL was separated from plasma by size-exclusion chromatography and isolated from pooled fractions, using a lipid binding resin. HDL bound proteins from each subject were analyzed by electrospray ionization tandem mass spectrometry and a spectral index value (derived from normalized spectrum counts and the number of subjects in which the peptide was detected) was used to examine the association between the HDL proteome and atherosclerosis severity. This analysis identified changes in 14 out of the total of 123 identified proteins. Of those that changed, some displayed clear correlations with disease severity, whereas others were only affected in the severe disease group. Some of these proteins have been previously suggested to play a role in the interaction of HDL and atherosclerosis, such as apoA-IV (+20%; p<0.02) and paraoxonase-1 (-21%; p<0.05). Others may represent new HDL associated biomarkers, such as Ig lambda-3 chain C region (+188%; p<0.01). We are currently developing a novel approach for the analysis of HDL proteome data, which will take into account all identified proteins as a single proteomic signature. This signature may have potential as a simplified metric of HDL composition with a direct connection to functionality.
Abstract 157: Alternative Splicing of PLD1 and Its Role in Modulating Inflammatory Phenotypes in Primary Human Macrophages
Authors: Lin Jennie , Cieply Benjamin , Zhang Hanrui , Barash Yoseph , Gerelus Mark , Reilly Muredach P , Rader Daniel J , Musunuru Kiran
Published: June 2021

With roles in homeostasis and immunity, human macrophages can readily shift phenotype across the pro-inflammatory M1 and reparative M2 spectrum in response to environmental challenges, but the mechanistic underpinnings of this process remain incompletely understood. Recently we showed from RNA-seq that hundreds of alternative splicing events occur during primary human macrophage LPS / IFN-γ activation to the M1 phenotype associated with cardiometabolic diseases including atherosclerosis. From these events, we found that in M1 activation but not M2 activation, PLD1, which encodes an enzyme catalyzing the hydrolysis of phosphatidylcholine, undergoes increased inclusion of exon 16, with an increased percent spliced in (PSI) of 17.6% calculated from RNA-seq reads and validated by semi-quantitative PCR in human macrophages derived from 3 independent donors (A). Based on these observed differences, we hypothesized that PLD1 has important functional roles in M1 activation. To test this, we used siRNAs to knock down PLD1 in primary human macrophages (B), which induced an increased inflammatory response to LPS / IFN-γ stimulation. This increased inflammatory response was consistent with characteristics of a more robust M1 phenotype, including decreased zymosan phagocytosis in both resting and stimulated macrophages (C), and increased secretion of inflammatory cytokines such as IL-1β, IL-18, IL-6, and CCL5 (D, E). Morpholino induced skipping of PLD1’s exon 16 reduced PLD enzymatic activity to a similar degree seen in morpholino inhibition of PLD1 translation (F, G). These findings suggest that PLD1 may be a novel modulator of M1 activation in an isoform-dependent manner.