bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P138:
Schistosoma Mansoni-
Induced Shedding Of Extracellular Vesicles From Lung Microvascular Endothelial Cells
Authors: Oliveira Suellen D , Schwartz Andrew , Williams David , Bonini Marcelo , Silva Claudia Lucia Martins , Minshall Richard D
Published: May 2022

Introduction:
Schistosomiasis-associated Pulmonary Arterial Hypertension (Sch-PAH) is a life-threatening complication of chronic
Schistosoma mansoni
infection. Sch-PAH is marked by obliteration and remodeling of the pulmonary vasculature in response to
S. mansoni
eggs. Remodeled and occluded vessels increase vascular resistance and can lead to heart failure and death. Currently, there are no target therapies for the treatment of Sch-PAH.

Hypothesis:
S. mansoni
egg-derived antigens alter pulmonary endothelial cell (EC) Caveolin-1 (Cav-1) expression and phosphorylation contributing to the loading and shedding of extracellular vesicles (EVs), which in turn stimulates abnormal EC survival leading to Sch-PAH.

Methods/Results:
Immunohistology and western blot analysis of lungs from
S. mansoni
-infected mice revealed a significant decrease in Cav-1 expression in the egg-dependent granuloma area when compared to uninfected control animals, indicating that local
S. mansoni
-associated molecules and inflammatory mediators contribute to Cav-1 depletion. Previously we showed that upon vascular injury, Cav-1 deficiency leads to the survival of an abnormal EC phenotype through an unclear mechanism. Analysis of survival-associated genes in isolated lung ECs from Flk1
+/GFP
;
Cav1
-/-
mice, revealed a high expression of the anti-apoptotic BIRC2 and BIRC5 (known as c-IAP2 and surviving, respectively) when compared to control ECs. Interesting, in vitro exposure to the major
S. mansoni
egg antigen, Sm-p40 (1 μg/mL), induced time-dependent phosphorylation of Cav-1 Tyr14 in human lung microvascular ECs (HMVEC-L; 228.2 +/- 38.28% of control; p<0.05; n=4), which culminated in c-IAP2 expression. Furthermore, Sm-p40 treatment of HMVEC-L increased ERK1/2 phosphorylation, which in presence of pro-inflammatory mediators IL-6, TNF-a, and ATP, significantly increased the shedding of EVs implicating P-ERK1/2 in the mechanism of shedding of Cav-1-EVs.

Conclusions:
This work is uncovering the biological role and potential translational relevance of pathogen-induced EC-Cav-1 depletion via shedding of EVs during Sch-PAH.
Abstract P144: Bariatric Surgery Normalizes Diabetes Risk Index By 1 Month Post-operation
Authors: Sinatra Vincent J , Lin BingXue , Parikh Manish , Berger Jeffrey S , Fisher Edward A , Heffron Sean P
Published: May 2022

The Diabetes Risk Index (DRI) is a composite of NMR-measured lipoproteins and branched chain amino acids shown to be predictive of diabetes in prospective cohorts. Bariatric surgery is indicated in patients with severe obesity, many of whom are at high-risk for developing diabetes. Substantial weight loss occurs after bariatric surgery and sustained weight loss likely contributes to lower risk of diabetes and cardiovascular disease. However, evidence suggests that bariatric surgical procedures themselves may contribute to reducing risk independent of weight loss. We aimed to investigate DRI and its association with weight loss over one year following bariatric surgery. We examined 51 severely obese premenopausal women without diabetes, 25 of whom met criteria for metabolic syndrome. DRI, BMI, body weight and waist measurements were made before and at 1, 6 and 12 months after Roux-en-Y Gastric Bypass or Sleeve Gastrectomy. Values were compared to healthy women with normal BMI (18.5 - 24.9 kg/m
2
; n=15). Obese (BMI 44.7 ± 6.2 kg/m
2
) subjects had significantly elevated DRI scores prior to surgery (35 [26, 39] vs 12 [1, 20]; p <0.0001). At 1 month after surgery, BMI had decreased 5.1 ± 1.1 kg/m
2
, but DRI decreased so that it no longer differed from that of normal BMI controls (Figure). Subjects continued to lose weight, whereas DRI remained similar, throughout 12 months. Changes in DRI did not correlate with changes in BMI, body weight or waist circumference at any time during follow-up (r = -0.16 - 0.16). There was no difference in response between surgeries or pre-operative metabolic syndrome status. Our analysis of DRI scores supports the capacity of bariatric surgery to reduce risk of developing diabetes in severely obese individuals with or without metabolic syndrome. The rapidity of normalization and the lack of associations with weight loss or anthropometric changes suggest that bariatric surgical techniques may have inherent effects that improve cardiometabolic risk.
Abstract P152: Downregulation Of Tissue-nonspecific Alkaline Phosphatase In Endothelial Cells Does Not Affect Calcification But Reduces Cardiac Function In Mice With Atherosclerosis
Authors: Than Sandy , Canellos Maria , Shamsian Ethan , Singh Mohnish , Savinova Olga V
Published: May 2022

Introduction:
We have previously demonstrated that overexpression of tissue-nonspecific alkaline phosphatase (TNAP) in endothelial cells increases vascular calcification and accelerates atherosclerosis. Whether TNAP activity in vascular endothelium is essential for intimal calcification and the progression of atherosclerosis is unknown.

Hypothesis:
We hypothesized that elimination of TNAP expression in endothelial cells would be sufficient to reduce intimal calcification associated with atherosclerosis.

Methods:
Endothelial TNAP knockout (KO) mice were developed on the background of the low density lipoprotein receptor mutation. Endothelial TNAP KO (n = 8) and control (n = 9) mice were fed a Western diet starting from 8 weeks and until 52 weeks of age. Cardiac structure and function were assessed by echocardiography. Calcium volume was measured in the aortic root and aortic arch area by microcomputed tomography (microCT). Data were analyzed using a 2-way ANOVA to calculate the effects of the genotype adjusted for sex.

Results:
MicroCT data did not show any statistically significant decrease in calcification in endothelial TNAP KO mice compared to controls; however, other cardiac parameters were affected by genotype. The physiological examination of the mice demonstrated that ablation of TNAP expression in the endothelium resulted in significant reduction in ejection fraction, stroke volume, and cardiac output in the absence of cardiac hypertrophy.

Conclusion:
Contrary to our hypothesis, TNAP has no effect on natural progression of calcified atherosclerosis; however, our data indicate that TNAP is required for protection of cardiac function under conditions of atherosclerosis in mice
Abstract P153: Upregulation Of Thrombospondin-1 Associates With Accelerated Atherosclerosis And Reduced Smooth Muscle Cell Differentiation In Metabolic Syndrome
Authors: Gupta Shreya , Khanal Saugat , Mathias Amy , Lallo Jason , Ferrell Jessica , Raman Priya
Published: May 2022

Hyperglycemia and obesity, characteristic of metabolic syndrome (MetS), are important risk-factors for atherosclerosis. MetS patients manifest increased vascular smooth muscle cell (VSMC) migration and proliferation, hallmark of VSMC phenotypic transition, critical for evolution of atherosclerosis. We previously reported that high glucose and high leptin independently upregulate a potent proatherogenic matricellular protein, Thrombospondin-1 (TSP-1), expression in VSMC. The goal of the present study was to interrogate the role of TSP-1 in SMC de-differentiation and development of atherosclerosis in MetS. We utilized a mouse model of combined MetS and atherosclerosis (KKAy
+/-
/ApoE
-/-
) generated by crossing obese hyperglycemic agouti KKAy
+/-
mice with atherosclerotic ApoE
-/-
. Upon weaning (4 wks age), male yellow KKAy
+/-
/ApoE
-/-
and age-matched black KKAy
-/-
/ApoE
-/-
littermates on standard lab diet were monitored monthly for body weight and blood glucose. At 24 wks age, mice were harvested after overnight fasting; plasma, aorta and heart were collected for biochemical, molecular and lesion studies. Prior to harvest, mice were subjected to treadmill exercise test at an incline with progressively increasing speed until exhaustion. At 16 wks age, yellow KKAy
+/-
/ApoE
-/-
mice showed significant increase in body weight and random blood glucose levels (<0.0001) vs. black KKAy
-/-
/ApoE
-/-
littermates, validating the MetS phenotype. Peak oxygen consumption (VO
2max
), maximum running speed and total run time until exhaustion were significantly reduced in MetS KKAy
+/-
/ApoE
-/-
mice (p<0.003 vs. non-MetS KKAy
-/-
/ApoE
-/-
), suggesting impaired cardiovascular fitness. Aortic root morphometry revealed 4-fold increase in lesion lipid burden in MetS vs non-MetS mice. This was accompanied with reduced LMOD (SM contractile marker) and SRF (transcriptional activator of SM contractile genes) expression in aortic vessels of MetS mice (p<0.04 vs. non-MetS mice). Notably, lesion abundance and reduced SMC differentiation associated with increased TSP-1 expression in the aortic vasculature of MetS mice. Together, our data suggest a putative role of TSP-1 in SMC de-differentiation and atherosclerotic lesion formation in metabolic syndrome.
Abstract P161: Novel Micro-CT Method To Assess Atherosclerotic Calcification In Mice
Authors: Kaur Jashandeep , Carpo Beatrice , Kosciuszek Nina D , Singh Mohnish , Savinova Olga V
Published: May 2022

Introduction:
Animal models are used to study mechanisms and consequences of calcification in atherosclerosis. As a standard practice, vascular calcification in a mouse model is measured by laborious histological preparation and calcium staining. In this study, we developed an
ex-vivo
micro-CT (μCT) method to quantify calcium volume in the aortic roots and arches in mice with atherosclerosis.

Objective:
The purpose of this study was to evaluate feasibility and compare calcium measurements between the μCT and histological methods.

Methods:
We used a mouse model of increased atherosclerotic calcification due to overexpression of tissue nonspecific alkaline phosphatase (TNAP) in macrophages. Formalin-fixed tissues were dissected and slowly immersed in mineral oil. Samples were assembled in cryogenic vials pre-filled with mineral oil. Cotton was used to immobilize and space the samples. μCT images of whole hearts and aortic arches were collected at a 7.1 micron resolution. Multiple Deep Learning models, implemented in Dragonfly 2020.1 (ORS), were trained on one sample for image segmentation. Histological analysis was performed according to standard protocols.

Results:
Multiple preparations of hearts and aortic roots can be scanned in a single overnight experiment. A Deep Learning model of the U-Net5 architecture was most successful in the segmentation of soft tissues and calcifications, while excluding μCT artifacts. The data showed a good correlation between detection of calcium by μCT and histology. μCT analysis of samples from 1-year-old TNAP transgenic mice demonstrated a 4-fold increase in calcification volume compared to control mice.

Conclusion:
Our μCT method is feasible and, in some aspects, superior to the traditional histological analysis because it is less time-consuming and less prone to artifacts. μCT approach is non-destructive thus permitting analyses of samples by multiple analytical methods after removal of mineral oil.
Abstract P162: Leveraging Lysosomal Calcium Flux As A Strategy To Induce Autophagy-lysosomal Biogenesis In Macrophages
Authors: Jeong Se-Jin , Zhang Xiangyu , Yeh Yu-sheng , Rodríguez-Vélez Astrid , Razani Babak
Published: May 2022

Dysfunction in macrophage lysosomes leading to poor handling of atherogenic lipids, apoptotic cells, and other cytotoxic debris leads to progression of atherosclerotic plaque size and complexity. Stimulation of TFEB, the master transcriptional regulator of autophagy-lysosomal biogenesis, in macrophages has been shown to promote the degradative capacity of plaque macrophages with reductions in atherosclerosis. Thus, there is interest in harnessing this pathway for cardiovascular therapeutics. One of the mechanisms by which TFEB is activated and triggered to translocate to the nucleus is the release of calcium from the lysosomal lumen to the cytosol via specialized channels located at the lysosomal membrane including mucolipin 1, also known as the transient receptor potential cation channel mucolipin 1 (TRPML1). We find that TRPML1 is highly expressed in macrophages and is involved in a feedback loop by which TFEB positively regulates TRPML1 mRNA expression to further support long-term autophagy-lysosomal biogenesis. Utilizing a mammalian TRPML agonist, mucolipin synthetic agonist 1 (ML-SA1), we show that induction of lysosomal calcium release can robustly trigger TFEB nuclear translocation in macrophages. In turn, numerous downstream TFEB transcriptional targets mediating autophagy-lysosomal biogenesis are upregulated resulting in salutary effects including enhanced aggrephagy and clearance of cytotoxic p62-enriched protein aggregates, blunting of inflammasome activation and proinflammatory IL-1β production, and reductions in macrophage apoptosis. Taken together, our data support that stimulation of lysosomal calcium flux can be a novel strategy of enhancing TFEB and macrophage degradative capacity in atherosclerosis.
Abstract P170: Targeting Angiotensin Ii-induced Endoplasmic Reticulum Stress In Vsmcs To Reduce Pathological Vascular Amyloid Burden And Remodeling
Authors: Cicalese Stephanie M , Okuno Keisuke , Preston Kyle , Eguchi Satoru
Published: May 2022

Hypertension is a complex disorder and risk factor for cardiovascular disease, ultimately contributing to premature death. The societal burden of hypertension is enormous, therefore better understanding of molecular mechanisms underlying hypertension and associated pathological vascular remodeling is needed for mechanism-targeted therapy development. Endoplasmic reticulum (ER) stress occurs in cells under increased protein synthesis, Ca
2+
flux, or ROS generating environments- all of which are induced by Angiotensin II signaling in vascular cells. ER stress leads to the accumulation of misfolded proteins, and mediates cell death, fibrotic, and hypertrophic responses. The aim of this study was to target ER stress via chemical chaperone 3-hydroxy-2-naphthoic acid (3HNA) or genetic overexpression of chaperone glucose-regulated protein 78 (GRP78) to reduce protein aggregation and pathological response in VSMCs. Rat primary VSMCs were treated with 500 μM 3HNA or 30 MOI lacZ-GRP78 prior to stimulation with Ang II (100 nM). Protein synthesis assessed via puromycin incorporation revealed ER stress inhibition blocked Ang II associated protein synthesis in VSMCs, fold change relative to control of 1.7 (saline + Ang II) to 1.2 (3-HNA + Ang II) treated cells (P<0.05). GRP78 overexpression attenuated Ang II induction pre-amyloid oligomers (P<0.01) from 2.1 aggregates/cell to 0.9 aggregates/cell. Proteomic assessment of aggregates revealed Ang II induced crystallin AB (1.3 fold rel to control), HSP70 (1.716 fold rel to control) and annexin a2 (1.3 fold increase) enrichment in detergent insoluble fractions, which was attenuated by GRP78 overexpression (P=0.03, 0.03, and 0.024, respectively). In a 2 week Ang II infusion model, 3HNA was injected in C57/Bl6 mice. Ang II-induced medial thickness and cardiac vessel fibrosis was significantly reduced by 3-HNA (P<0.05). SM22α Cre GRP78 mice were generated and found protective against Ang II induced cardiac hypertrophy via HW/BW ratio (Cre- vs Cre+ Ang II P<0.05), and a trend in reduction of aortic medial thickness (P=0.098). Overall, GRP78 chaperone expression reduced protein aggregation and subsequent vascular remodeling induced by hypertensive stimulus and may elude novel therapeutic targets.
Abstract P171: Glucose Consumption Of Vascular Cell Types In Culture; Toward Optimization Of Experimental Conditions
Authors: Torimoto Keiichi , Okuno Keisuke , Kuroda Ryohei , Cicalese Stephanie M , Eguchi Kunie , Elliott Katherine , Kawai Tatsuo , Eguchi Satoru
Published: May 2022

Any experimental outcomes are potentially influenced by extracellular glucose availability and its cellular metabolism. However, there is a lack of attention paid to the supply and utilization of glucose in many cultured experiments. Surveillance of vascular related journals for the past 5 years demonstrated that less than 20% of published articles declared the medium glucose concentration. The present studies were designed to seek ideal medium glucose concentration(s) in various cell types with particular attention paid to changes in glucose consumption. By using distinct glucose concentrations in Dulbecco's Modified Eagle's medium (DMEM), we identified ideal glucose media for stimulation experiments. We have compared glucose consumption in three vascular cell types, endothelial cells (EC), vascular smooth muscle cells (VSMC) and adventitial fibroblasts (AF) with or without angiotensin II (AII) stimulation. In all cell types after a 48h incubation in relatively low glucose media (1 g/L), medium glucose concentration was reduced to almost 0 (EC 0.01±0.01 g/L, VSMC 0.13±0.05 g/L, AF 0.02±0.01 g/L). Whereas medium glucose concentration remained significantly higher (EC 2.77±0.05 g/L, VSMC 3.87±0.05 g/L, AF 3.32±0.01 g/L) when cells were incubated for 48h in high glucose (4.5 g/L) media. In middle glucose (2.75 g/L) media, medium glucose concentration remained in physiological ranges (EC 0.62±0.18 g/L, VSMC 1.98±0.07 g/L, AF 1.17±0.17 g/L). AII treatment enhanced glucose consumption in AF and low passage VSMC but not in EC. In AF, AII induction of target proteins varied depending on the glucose concentration used. In low glucose media induction of Grp78 or hexokinase II was highest, whereas induction of VCAM1 was lowest. Utilization of specific inhibitors further suggest essential roles of AT1 receptor and glycolysis in AII-induced fibroblast activation. Overall, the present study demonstrates a high risk of hypo- or hyperglycemic conditions when standard low or high glucose media is used with vascular cells. Moreover, these conditions may significantly alter experimental outcomes. Medium glucose concentration should be monitored during any culture experiments and utilization of middle glucose media is recommended for all vascular cell types.
Abstract P172: Pulmonary Valve Papillary Fibroelastoma In A Patient With Chronic Chest Pain
Authors: Karimi Hosseini Davood , Ahmed Noreen , Shkolnik Elina , Philip Lesley , Sandhu Prabhjeet , Yildiz Ali , Patel Rajiv , Villegas Dennis
Published: May 2022

Primary cardiac tumors are rare entity, and the most common benign cardiac neoplasms include myxomas, lipomas and papillary fibroelastomas (PFE) with PFE accounting for 8% of the tumors. A 53-year-old female presented with chronic chest pain and exertional shortness of breath. She first sought evaluation four years ago, when her pulmonologist referred her to thoracic surgery after an incidental finding of a 3.9 cm anterior mediastinal mass on CT-scan (Figure 1A-B). The patient described the pain as a constant pressure and squeezing sensation in her chest and worsening shortness of breath while laying on her sides, relieved only when a pillow was propped up under her chest. The pain did not significantly reduce her mobility or capacity to work. Initial evaluation, including transthoracic echocardiogram, stress testing, carotid ultrasound, and pulmonary function testing - was performed and found to be unremarkable. Further evaluations revealed a hiatal hernia, leading her symptoms to be attributed to GERD as causing her persistent non-exertional chest pain. The patient remained in good health despite her symptoms persisting. She later re-sought attention for chest pain on exertion and hoarseness three years later. She again underwent a full cardiopulmonary workup, yielding benign findings on EKG, troponins, and echocardiogram. Due to no alarming symptoms at the time, she was subsequently discharged. The patient continued to endorse chest pain and a follow-up MRI revealed a now 5 cm anterior mediastinal mass, thought to be a thymic cyst. Over the next few months, the patient continued to endorse worsening chest pain and exertional shortness of breath. She followed up with her cardiologist and a subsequent TEE was significant for a pulmonic valvular mass. She was admitted to the hospital for further investigation. During her hospital stay, the patient underwent surgery to resect the anterior mediastinal mass likely to be a thymic cyst and the pulmonary artery mass which was determined to be a pulmonary valve papillary fibroelastoma in the histopathology exam (Figure 2). The surgery was successful with no complications postoperatively. The patient was discharged with no further chest pain and with improvement of the exertional shortness of breath.
Abstract P174: Nuclear Smooth Muscle α-actin Is Critical For Smooth Muscle Cell Differentiation And To Prevent Cerebrovascular Disease
Authors: Kwartler Callie , Duan Xue-yan , Ma Shuangtao , Kaw Anita , Kernell Caroline , Wang Charis , Chen Jiyuan , Shen Xuetong , Milewicz Dianna M
Published: May 2022

Missense mutations in ACTA2, encoding α-smooth muscle actin (α-SMA), predispose to thoracic aortic aneurysms. A subset of these ACTA2 mutations also predispose to childhood onset cerebrovascular disease characterized by occlusion of the distal internal carotid arteries and small vessel disease. In our studies to identify how specific ACTA2 mutations predispose to cerebrovascular disease, we confirmed that α-SMA translocates to the nucleus in wildtype (WT) smooth muscle cells (SMCs), is enriched in the nucleus over β-actin with differentiation of SMCs, associates with the INO80 chromatin remodeling complex, and selectively binds to the promoters of SMC contractile genes. Expression of individual mutant α-SMAs in 293 cells determined that missense mutations associated with cerebrovascular disease inhibit this nuclear translocation. To further examine the effects of reduced nuclear α-SMA with ACTA2 mutations, we used two model systems: pluripotent stem cell-derived SMCs from patients with ACTA2-associated cerebrovascular disease (ACTA2 R179C) as well as SMCs explanted from a knockin mouse model (Acta2SMC-R179C/+). Both mouse and human cells harboring the R179C mutation proliferate and migrate more than WT SMCs, are less differentiated than WT SMCs, and have increased expression of pluripotency-associated genes. Although the mutation is heterozygous, these cells show a dominant negative impact on nuclear α-SMA function through dramatically reduced levels of α-SMA in the nucleus, in the INO80 chromatin remodeling complex, and on the promoters of SMC-specific genes. Finally, forced nuclear localization of α-SMA in WT SMCs increases levels of SMC contractile proteins. Taken together, we have identified a novel role for α-SMA in driving differentiation of SMCs, and our data supports that defects in this nuclear role drive cellular phenotypes consistent with the cerebrovascular disease seen in patients with ACTA2 R179 mutations.
Abstract P176: Pharmacological Inhibition Of Drp1 Dependent Mitochondrial Fission Alleviates Mitochondrial Dysfunction-associated Cell Death In Vascular Smooth Muscle Cells
Authors: Jadli Anshul , Gomes Karina , Patel Vaibhav B
Published: May 2022

Background:
Vascular smooth muscle cell (VSMC) death has been linked to the pathogenesis of various vascular diseases. Excessive mitochondrial fission in response to pathological stimuli contributes to apoptosis, thus inhibition of mitochondrial fission appears to be a promising therapeutic target in vascular disease. Mitochondrial division inhibitor 1 (mDivi-1) inhibits Drp1 dependent fission by attenuation of its GTPase activity. The limited literature on mDivi-1-mediated attenuation of VSMCs apoptosis and its effect on the pathogenesis of vascular disease warrants further investigation.

Methods and Results:
Staurosporine (STS)-induced apoptosis and attenuated cell proliferation were observed in VSMCs. The treatment with mDivi-1 ameliorated these detrimental effects of STS and led to reduced apoptosis, and increased cell proliferation. The mitochondrial fission was evaluated using Mitotracker Red staining which showed reduced mitochondrial fragmentation with significantly higher mitochondrial networks, mean branch length, and mitochondrial footprint upon treatment with mDivi-1. Drp1 levels were increased in STS-treated cells with localization in the mitochondria. Cellular and mitochondrial stress response assessed by DHE and mitosox staining. mDivi-1 treatment attenuated STS-induced ROS generation in VSMCs. Mitochondrial function via evaluation of Mitochondrial permeability transition pore (mPTP) opening, mitochondrial membrane potential, and bioenergetic profile showed significantly reduced mPTP opening and higher metabolic potential represented by OCR and ECAR in mDivi-1-treated cells compared to STS group. The autophagy marker LAMP1 showed increased colocalization in mitochondria of STS-treated VSMCs. The apoptotic proteins were assessed using a protein array. The mDivi-1 treated VSMCs showed significantly reduced autophagy and apoptosis-related proteins. This suggested attenuation of apoptosis-mediated mitochondrial damage upon treatment with mDivi-1.

Conclusion:
Pharmacological inhibition of mitochondrial fission provides a potential therapeutic target in VSMCs death-associated vascular diseases.
Abstract P178: A Novel Subset Of Adventitial Vascular Stem Cells Paracrine Control Media Smooth Muscle Cell Dedifferentiation Leading To Intimal Hyperplasia In Vein Grafts
Authors: Wu Weiwei , Yin Rong , Tan Wenbin , Nagarkatti Mitzi , Nagarkatti Prakash , Sanez-Garcia Isidro , Cai Guoshuai , Robinson Igor B , Cui Taixing
Published: May 2022

Vein graft failure (VGF) is associated with vein graft (VG) intimal hyperplasia, which is characterized by abnormal accumulation of vascular smooth muscle cells (SMCs). The majority of neointimal SMCs are derived from pre-existing vascular SMCs via a process of vascular SMC dedifferentiation; however, the underlying mechanisms remain poorly understood. Here, we tracked down the fate of vascular stem cells (VSCs) expressing stem cell antigen 1 (SCA1) in VG remodeling. After transplantation, most of the donor venous cells including endothelial cells, SMCs, and VSCs died within 3 days, and the recipient arterial SCA1+ VSCs were recruited to repopulate the adventitia and the intima but did not differentiate into neointimal SMCs in VGs. However, ablation of the SCA1+ VSCs ameliorated intimal hyperplasia in VGs. Single-Cell RNA sequencing (scRNA-seq) analysis revealed a unique subset of VSCs expressing Sca1 and cyclin-dependent kinase 8 (Cdk8) in the artery. Notably, the number of SCA1+CDK8+ cells was increased in the adventitia prior to dramatic proliferation of SMCs in the media and neointima in VGs. Inactivation of CDK8 intensified SCA1+ VSC naïve stemness but suppressed SCA1+ VSC proliferation, migration, and exosome release for paracrine enforcement of vascular SMC dedifferentiation. A short-time perivascular delivery of CDK8 inhibitors ameliorated adventitial SCA1+ VSC accumulation associated with a long-term efficacy in suppressing intimal hyperplasia in VGs. There findings uncover that a novel subset of adventitial VSCs paracrine control media SMC dedifferentiation for intimal hyperplasia in VGs toward VGF.
Abstract P180: Tunable Elastin-like Protein-based Hydrogels For Cell Transplantation In Vascular Repair
Authors: Shayan Mahdis , Suhar Riley , Huang Ngan F , Heilshorn Sarah C
Published: May 2022

Introduction:
Endothelial cells can improve blood perfusion in diseased blood vessels; however, direct injection of cells significantly decreases their survival and functionality for angiogenesis. To address these limitations, we study a family of engineered extracellular matrices with tunable biochemical and biomechanical cues for enhanced survival and improved angiogenic behavior of ECs.

Materials & Methods:
Engineered hydrogels, termed ELP-PEG, consists of two components of a hydrazine-modified elastin-like protein (ELP-HYD) and an aldehyde- or benzaldehyde-modified, polyethylene glycol (PEG-ALD or PEG-BZA), which interact with each other through hydrazone dynamic covalent chemistry (DCC) bonds to form ELP-PEG hydrogels. Stiffness is controlled by altering the number of PEG-ALD or PEG-BZA crosslinks, and the stress relaxation rate is tuned by varying the PEG-ALD crosslinks vs. PEG-BZA. Stiffness and stress relaxation rates of the hydrogels were assessed by dynamic oscillatory rheology. Afterward, human umbilical vein endothelial cells (HUVECs) were encapsulated within gels to assess cell viability and spreading using a Live/Dead Cytotoxicity assay and confocal fluorescence imaging.

Results and Discussion:
Stress relaxation rate in ELP/PEG-ALD is much slower compared with a similar combination in ELP/PEG-BZA. Rheology measurements of the RGD-ELP/PEG (2%/2%) hydrogels demonstrated a storage modulus of 800Pa. It confirms the tunability of the stress relaxation rate with constant stiffness. Cell viability assay demonstrated that both hydrogels could support high cell viability (>90%) for 7 days. After 7 days, cell spreading increased in RGD-ELP/PEG-BZA hydrogels, however, cells did not form elongated morphology in RGD-ELP/PEG-ALD hydrogels, suggesting that stress relaxation rate and mechanical stiffness are key characteristics in modulating endothelial cell behavior.

Conclusions:
ELP/PEG-ALD/BZA promotes the angiogenic behavior of endothelial cells and is a promising candidate for cell delivery in vascular diseases.
Abstract P181: Machine Learning Methods For Predicting 30-day All Cause Readmission Following Carotid Endarterectomy Among Acute Ischemic Stoke Cases: A Nsqip Study (2014 - 2017)
Authors: Saxena Anshul , Rubens Muni , Ramamoorthy Venkataraghavan , Suarez Mariana , Appunni Sandeep , McGranaghan Peter , Veledar Emir , Garelnabi Mahdi O
Published: May 2022

Background:
Carotid endarterectomy (CEA) is associated with improved overall clinical outcomes in patients with acute ischemic stroke (AIS). However, studies on rates and factors associated with readmission following CEA for AIS are scarce. In this study, we used machine learning (ML) methods to identify the factors associated with readmission using a large-scale national database.

Methods:
We used National Surgical Quality Improvement Program (NSQIP) registry (2014-2017) and included patients 18 years or older, who underwent CEA for AIS. AIS and CEA were identified using ICD-9 and ICD-10 diagnosis and CPT procedure codes, respectively. We used Naïve Bayes, Boosted Decision Trees, and Bootstrapped Random Forest classification techniques to explore the predictors of 30-day readmission using demographics, past medical history, and preoperative variables.

Results:
There were a total of 22,373 AIS patients who underwent CEA. Mean (SD) age of the patients was 70.7 (9.4) years, and 61% were men. Majority were non-Hispanic White (80%), followed by non-Hispanic Black (4.6%). During the study period, 1 in 15 AIS patients who underwent CEA experienced 30-day readmission. Bootstrapped Random Forest classification performed best and Naïve Bayes worst with an AUROC of 92% and 59% respectively. The top 5 predictors of 30-day readmission after CEA were Hematocrit, BUN, Creatinine, WBC count, and Platelet count, all collected pre-operatively.

Conclusion:
Our study showed that ML techniques could accurately predict 30-day readmission using pre-operative risk factors. This ML model could be incorporated in EMR as a potential clinical decision support system. Implementing this system could help in early identification of patients who are at high risk for readmission following CEA. This could help physicians to plan and intervene effectively and prevent short-term readmissions; thereby improving quality of care and saving healthcare costs.
Abstract P184: Novel Model Of Ionizing Radiation-induced Mouse Coronary Arteriosclerosis, Which Was Attenuated By Precise Time Treatment Of Poly (adp-ribose) Polymerase (parp) Inhibitor
Authors: Ko Kyung ae , Abe Jun-ichi , Kotla Sivareddy , GI YOUNG JIN , Imanishi Masaki , Fujiwara Keigi
Published: May 2022

Background:
It is well known that radiation therapy (RT) induces coronary artery disease (CAD). However, since we don’t have an adequate mouse model to evaluate CAD after RT, it has been difficult to perform pre-clinical study to detect the effective treatment. The inhibition of PARPs can prevent apoptosis and inflammation. Although PARP inhibitors have already been used as anti-tumor agents, their long-term use in CAD is not recommended because the inhibition of DNA damage response can cause DNA instability and eventually cancer. In this study, we restricted the use of PARP inhibitors to the time of IR exposure only and detected IR-induced atherosclerosis (AS).

Methods and Results:
Marino et al. reported that thoracic aortic constriction (TAC) can exacerbate coronary artery stenosis and myocardial infarction in ApoE
-/-
mice. We tested this model in LDLR
-/-
mice that were fed a high fat (HFD) with or without IR (3, 5, and 10 Gy). We observed cardiac hypertrophy and dose-dependent reduction of fractional shortening (FS) after 4-5 weeks of TAC (FS%, 33.74 ± 7.35 (Non-IR, n=9), 26.59 ± 9.19 (3Gy, n=6), 28.49 ± 2.49 (5Gy, n=5), 22.62 ± 6.16 (10Gy, n=6), mean ± SD, p <0.05). Most importantly, we observed localized cardiac dysfunction and infarct only in mice exposed to 10 Gy (n = 2 out of 6), detected by transmural strain analysis with echocardiography. Next, the whole heart was sectioned, with sets of 11 consecutive sections of 5 μm collected every 450 μm interval. We also found the diffuse increase of vascular wall thickness at left anterior descending coronary artery in mice exposed to IR. We fed a HFD on LDLR
-/-
mice with Olaparib (10 mg/kg/day) or vehicle one day before & after, and the day of IR (5 Gy twice, total 6 days only), then performed TAC. The reduction of FS induced by IR (10 Gy) was significantly improved by the precise time treatment of Olaparib against IR.

Conclusion:
These data suggested the usefulness of TAC-induced coronary AS mouse model to develop medical countermeasures against RT-induced CAD as pre-clinical study.
Comparisons of Prehospital Delay and Related Factors Between Acute Ischemic Stroke and Acute Myocardial Infarction
Authors: Yoon Cindy W. , Oh Hoonji , Lee Juneyoung , Rha Joung‐Ho , Woo Seong‐Ill , Lee Won Kyung , Jung Han‐Young , Ban Byeolnim , Kang Jihoon , Kim Beom Joon , Kim Won‐Seok , Yoon Chang‐Hwan , Lee Heeyoung , Kim Seongheon , Kim Sung Hun , Kang Eun Kyoung , Her Ae‐Young , Cha Jae‐Kwan , Kim Dae‐Hyun , Kim Moo‐Hyun , Lee Jang Hoon , Park Hun Sik , Kim Keonyeop , Kim Rock Bum , Choi Nack‐Cheon , Hwang Jinyong , Park Hyun‐Woong , Park Ki Soo , Yi SangHak , Cho Jae Young , Kim Nam‐Ho , Choi Kang‐Ho , Kim Yongcheol , Kim Juhan , Han Jae‐Young , Choi Jay Chol , Kim Song‐Yi , Choi Joon‐Hyouk , Kim Jei , Jee Sung Ju , Sohn Min Kyun , Choi Si Wan , Shin Dong‐Ick , Lee Sang Yeub , Bae Jang‐Whan , Lee Kunsei , Bae Hee‐Joon
Published: May 2022

Background

Prehospital delay is an important contributor to poor outcomes in both acute ischemic stroke (AIS) and acute myocardial infarction (AMI). We aimed to compare the prehospital delay and related factors between AIS and AMI.

Methods and Results

We identified patients with AIS and AMI who were admitted to the 11 Korean Regional Cardiocerebrovascular Centers via the emergency room between July 2016 and December 2018. Delayed arrival was defined as a prehospital delay of >3 hours, and the generalized linear mixed‐effects model was applied to explore the effects of potential predictors on delayed arrival. This study included 17 895 and 8322 patients with AIS and AMI, respectively. The median value of prehospital delay was 6.05 hours in AIS and 3.00 hours in AMI. The use of emergency medical services was the key determinant of delayed arrival in both groups. Previous history, 1‐person household, weekday presentation, and interhospital transfer had higher odds of delayed arrival in both groups. Age and sex had no or minimal effects on delayed arrival in AIS; however, age and female sex were associated with higher odds of delayed arrival in AMI. More severe symptoms had lower odds of delayed arrival in AIS, whereas no significant effect was observed in AMI. Off‐hour presentation had higher and prehospital awareness had lower odds of delayed arrival; however, the magnitude of their effects differed quantitatively between AIS and AMI.

Conclusions

The effects of some nonmodifiable and modifiable factors on prehospital delay differed between AIS and AMI. A differentiated strategy might be required to reduce prehospital delay.
Studies on the subfamily Exothecinae (Hymenoptera: Braconidae) with the description of a new genus and a new species from India
Authors: SINGH LONGJAM RONI KUMAR , ACHTERBERG C. VAN , SHEELA S.
Published: May 2022

A new genus of the subfamily Exothecinae, Occipitotus Singh & van Achterberg gen. nov. is proposed. The type species Occipitotus langpramensis Singh & van Achterberg sp. nov., is described and illustrated, which is the second species of the subfamily known from India. A key to the extant genera of the subfamily Exothecinae is provided.
How Do Regional Distributions of Daily Precipitation Change under Warming?
Authors: Chadwick Robin , Pendergrass Angeline G. , Alves Lincoln Muniz , Moise Aurel
Published: May 2022

Global warming is changing the intensity distribution of daily precipitation, with an increased frequency of heavy precipitation and reduced frequency of light/moderate precipitation in general circulation model (GCM) projections. Projected future CMIP5 GCM changes in regional daily precipitation distribution can be described by a combination of two idealized modes: a frequency decrease mode, representing a reduction in the frequency of precipitation at all rain rates; and a frequency shift mode, where the distribution shifts toward heavier rain rates. A decrease in daily precipitation frequency and an increase in intensity are projected in most regions, but the magnitude of change shows large regional variations. The two modes generally capture the projected shift from light/moderate to heavy rain rates but do not recreate GCM changes at the very highest and lowest rain rates. We propose a simple framework for deep convective precipitation change based on the dry static energy (DSE) budget, which provides a physical explanation of these idealized modes in regions and seasons where deep convection dominates precipitation. One possibility is that a frequency decrease mode is driven by increased convective inhibition (CIN). In this DSE framework, increased moisture under warming could influence the shape of the precipitation intensity distribution, particularly at the highest rain rates, but does not govern the overall magnitude of the shift to heavier rain rates, which is not well described by the Clausius–Clapeyron relationship. Changes in daily regional precipitation are not free to respond only to local changes (in e.g., moisture) but are also constrained by the DSE budget, particularly by DSE transport associated with the large-scale circulation.
The Kalman Filter as Post-Processor for Analog Data–Model Assimilation in Paleoclimate Reconstruction
Authors: Wahl Eugene , Zorita Eduardo , Hoell Andrew
Published: May 2022

We present an offline paleo-data assimilation methodology that formally combines the analog assimilation method (AA) and the Kalman filter (KF), utilizing the KF as a postprocessor of the AA output. This methodology can be applied to reconstruct climate fields that are spatially separated from proxy-based reconstructions by using the spatial covariability generated by a climate model. Our method is applied to a set of spatially resolved and spatially consistent climate reconstructions of several variables reflecting different seasons, incorporating the application of methodological variants that have undergone rigorous testing in terms of both improving statistical methodology and physical interpretation. This contrasts with applications primarily based on transfer relationships of annual means of local, single variable or bivariate, climate model priors into paleo proxy states. The gains from adding the KF postprocessor are modest in our test case of reconstructing sea level pressure (SLP) geopotential height fields in the northeast Pacific, utilizing paleoclimatic temperature and moisture reconstructions in western North America. Notably, SLP reconstruction skill is enhanced in the oceanic region south of Alaska that is strongly associated with wet winters in western North America. The results suggest that the AA method is approaching optimality in this test case, driven by the quality of the paleoreconstruction information used to drive the AA process, along with the realism of the climate model employed, to which the KF postprocessing step is added. The derived reconstructions are then used for evaluation of the relationship between winter SLP and precipitation in California over the past ∼450 years.
An Improved Climatological Forecast Method for Projecting End-of-Season Water Requirement Satisfaction Index
Authors: Turner William A. , Husak Greg , Funk Chris , Roberts Dar A. , Jones Charles
Published: May 2022

A simple—yet powerful—indicator for monitoring agricultural drought is the water requirement satisfaction index (WRSI). In data-sparse, food-insecure areas, the WRSI is used to guide billions of dollars of aid every year. The WRSI uses precipitation (PPT) and reference evapotranspiration (RefET) data to estimate water availability relative to water demand experienced over the course of a growing season. If the season is in progress, to-date conditions can be combined with climatological averages to provide insight into potential end-of-season (EOS) crop performance. However, if the average is misrepresented, these forecasts can hinder early warning and delay precious humanitarian aid. While many agencies use arithmetic average climatologies as proxies for “average conditions,” little published research evaluates their effectiveness in crop-water balance models. Here, we use WRSI hindcasts of three African regions’ growing seasons, from 1981 to 2019, to assess the adequacy of the arithmetic mean climatological forecast—the Extended WRSI. We find that the Extended WRSI is positively biased, overestimating the actual EOS WRSI by 2%–23% in East, West, and southern Africa. The presented alternative combines to-date conditions with data from previous seasons to produce a series of historically realistic conclusions to the current season. The mean of these scenarios is the WRSI Outlook. In comparison with the Extended WRSI, which creates a single forecast scenario using average inputs that are not covarying, the WRSI Outlook employs an ensemble of scenarios, which more adequately capture the historical distribution of distribution of rainfall events along with the covariability between climate variables. More specifically, the impact of dry spells in individual years is included in the WRSI Outlook in a way that is smoothed over in the Extended WRSI. We find that the WRSI Outlook has a near-zero bias score and generally has a lower RMSE. In total, this paper highlights the inadequacies of the arithmetic mean climatological forecast and presents a less biased and more accurate scenario-based approach. To this end, the WRSI Outlook can improve our ability to identify agricultural drought and the concomitant need for humanitarian aid.
An Iterative Storm Segmentation and Classification Algorithm for Convection-Allowing Models and Gridded Radar Analyses
Authors: Potvin Corey K. , Gallo Burkely T. , Reinhart Anthony E. , Roberts Brett , Skinner Patrick S. , Sobash Ryan A. , Wilson Katie A. , Britt Kelsey C. , Broyles Chris , Flora Montgomery L. , Miller William J. S. , Satrio Clarice N.
Published: May 2022

Thunderstorm mode strongly impacts the likelihood and predictability of tornadoes and other hazards, and thus is of great interest to severe weather forecasters and researchers. It is often impossible for a forecaster to manually classify all the storms within convection-allowing model (CAM) output during a severe weather outbreak, or for a scientist to manually classify all storms in a large CAM or radar dataset in a timely manner. Automated storm classification techniques facilitate these tasks and provide objective inputs to operational tools, including machine learning models for predicting thunderstorm hazards. Accurate storm classification, however, requires accurate storm segmentation. Many storm segmentation techniques fail to distinguish between clustered storms, thereby missing intense cells, or to identify cells embedded within quasi-linear convective systems that can produce tornadoes and damaging winds. Therefore, we have developed an iterative technique that identifies these constituent storms in addition to traditionally identified storms. Identified storms are classified according to a seven-mode scheme designed for severe weather operations and research. The classification model is a hand-developed decision tree that operates on storm properties computed from composite reflectivity and midlevel rotation fields. These properties include geometrical attributes, whether the storm contains smaller storms or resides within a larger-scale complex, and whether strong rotation exists near the storm centroid. We evaluate the classification algorithm using expert labels of 400 storms simulated by the NSSL Warn-on-Forecast System or analyzed by the NSSL Multi-Radar/Multi-Sensor product suite. The classification algorithm emulates expert opinion reasonably well (e.g., 76% accuracy for supercells), and therefore could facilitate a wide range of operational and research applications.

Significance Statement

We have developed a new technique for automatically identifying intense thunderstorms in model and radar data and classifying storm mode, which informs forecasters about the risks of tornadoes and other high-impact weather. The technique identifies storms that are often missed by other methods, including cells embedded within storm clusters, and successfully classifies important storm modes that are generally not included in other schemes, such as rotating cells embedded within quasi-linear convective systems. We hope the technique will facilitate a variety of forecasting and research efforts.