bluuebunny/crossref_metadata_2025 huggingface dataset:
Design of proportional integral and derivative controller using particle swarm optimization technique for gimbal system
Authors: Ahmad Mohd Hafiez , Osman Khairuddin , Samsudin Sharatul Izah
Published: May 2022

This paper presents the development of an optimal proportional, integral and derivative (PID) controller for controlling camera gimbal on unmanned aerial systems (UAV). Three optimal controller improvements are obtained using the suggested particle swarm optimization (PSO) technique. The PSO algorithm is initially built and integrated with the PID controller to control the DC motor gimbal. Before comparing the performance of a DC motor with PSO-PID with a DC motor with Zeigler-Nichols controller, the impacts of iteration numbers are explored. Finally, bode analysis was conducted to validate the stability of the proposed PSO-PID controller. Simulation is conducted within the MATLAB environment to verify the system's performance in terms of settling time, steady-state error and overshoot. The simulation results show has a longer settling time (0.91656 sec) than the Ziegler-Nichols controller (0.14316 sec) but a shorter rising time (0.091686 sec) than the Ziegler-Nichols controller (0.00094 sec). Furthermore, the overshoot was lowered from 12.941% to 0.959% as a result. As a result, the suggested PSO-PID controller technique outperforms the Ziegler-Nichols controller in terms of overshoot and rise time. Further study will investigate the integration of other optimisation methodologies such as fuzzy logic for better performance.
Mobile Ad Hoc networks intrusion detection system against packet dropping attacks
Authors: Sbai Oussama , Elboukhari Mohamed
Published: May 2022

Due to the extreme lack of a stable infrastructure, also self-organization of network components, unpredictable network topologies, and the lack of a central authority for routing, security assurance in mobile ad hoc networks (MANETs) is an important and difficult challenge. Among the famous threat that MANETs suffer from: blackhole, grayhole, and selfishness attacks, because the target of these attacks is to drop packets and disturb the routing operation of the network. A scalable, reliable, and robust network intrusion detection system (NIDS) should be created to effectively combat these families of network layer routing assaults in order to offer high availability for MANETs. In this paper, we present a MANETs-IDS based on machine learning algorithm against blackhole, grayhole, and selfishness attacks with Ad Hoc on-demand distance vector (AODV) routing protocol (RFC 3561) and optimized link state routing (OLSR) potocol (RFC 3626), using ns-3 simulation platform. Our simulation took into consideration the density of the network and a random mobility model of nodes. The obtained experimental results show that the proposed detection algorithm reached very promoting performances (in term of accuracy, processing time, time to build the model, precision, recall, F-measure).
Hybridization of zero forcing-minimum mean square error equalizer in multiple input multiple output system
Authors: Akande Damilare O. , Adeyemo Zachaeus K. , Arowolo Olakunle O. , Oseni Olasunkanmi F. , Obanisola Oluwole O.
Published: May 2022

Transmission of high data rate over multipath environment is associated with many wireless applications. However, this transmission results in high delay which leads to ISI distortion and causes high error. The existing LE in MIMO systems such as ZF equalizer used in addressing this problem reduces the ISI distortion completely but results in noise amplification. Likewise, MMSE equalizer only reduces the noise but cannot eliminate ISI. Therefore, this paper proposed a hybrid LE for Wi-Fi over Weibull fading channel. The hybridized ZF-MMSE equalizer was developed using conventional ZF and MMSE equalizers. Digital transmitted signal propagated over Weibull fading channel is received at the receiver through multiple antennas and then combined using MRC. The combined signal is then equalized using hybrid ZF-MMSE to eliminate both the ISI and the amplified noise. The developed model was simulated in MATLAB software environment and evaluation was performed using BER and Pout. The results obtained revealed that the hybrid ZF-MMSE equalizer gave better performance over existing equalizers in a MIMO system. Therefore, the proposed equalizer will help improve the performance of the Wi-Fi technology.
A novel design of a reconfigurable power divider with stubs for mobile applications
Authors: Garah Messaoud , Kenane Elhadi , Djahli Farid
Published: May 2022

Nowadays, the world knows a huge revolution in telecommunications, which has led to the need to integrate many services, such as long-term evolution (LTE), 5G, wireless local area network (WLAN), and Bluetooth, in one receiving and transmitting device. In the same way, all active and passive microwave circuits must be compatible with the integration of services, including the power divider, which is the subject of our work. This passive component is used in several fields such as medicine, surveillance, radar systems and others applications. Wilkinson divider is one of the most prominent power dividers, it is classified as narrow-band power divider and it is a very useful component in mobile applications (2.4 and 5.8 GHz). In this work, a reconfigurable 4-ports Wilkinson divider is designed using PIN diodes and simulated using connected speech test (CST) simulator software. This designed divider can easily modify the operation mode by activating or disabling the PIN diodes.
Performance improvement on spectral efficiency and peak to avearge power reduction for 5G system
Authors: Thandapani Kavitha , Raju Ganesamoorthy , Pujali Maniganda
Published: May 2022

Filter bank multicarrier with offset quadrature amplitude modulation (FMBC/OQAM) system which can offer higher data rate 1 Gbps and improved bandwidth efficiency because of not require of cyclic prefix (CP) to avoid interference from the adjacent channels compared to fourth generation (4G) orthogonal frequency division multiplexing (OFDM) and other wireless systems. Because of these advantages FBMC/OQAM are used in fifth generation (5G) technology. In this FBMC/OQAM system, bandwidth capacity will be further increased by using multiple input multiple output (MIMO) channel. But in our proposed scheme we can use hyper–MIMO channel, in which at base station (BS) more number of transmitting antennas are used that for the improve the spectral efficiency performance better than traditional MIMO channel. But the most important drawback of hyper MIMO FBMC/OQAM is the higher peak to average-power ratio (PAPR) that degrade overall system performance in 5G. In 4G technology, to reduce PAPR a number of techniques are used. In our proposed system we can use mu-law non-linear companding technique and precoding discrete cosine transform (DCT) to reduce PAPR value for multicarrier modulation system compare to other systems.
An internet of things-based medication validity monitoring system
Authors: Makki Mohialden Yasmin , Mahmood Hussien Nadia , Jabbar Qabas Abdal Zahraa , Mohammed Mostafa Abduhgafoor , Sutikno Tole
Published: May 2022

Medicine is critical to our everyday lives and to the well-being of individuals of all ages and backgrounds. With the beginning of the Corona pandemic and a rise in Corona virus infection cases, the use of medications to prevent and recover from infection has increased, as well as to treat illness consequences, has grown. The effectiveness of medicines is greatly influenced by the expiration date. In this paper, a system for pharmacy or medical store's information storage system was developed and enhanced by automatically monitoring the validity of medications on a periodic basis and sending expiry reports to medicine authorities through e-mail to warn them that a medicine is approaching expiration. The system was also enhanced with internet of thing (IoT) for fast and secure delivery of the medicine validity report.
Clustering similar time series data for the prediction the patients with heart disease
Authors: Lafta Raid Luaibi , AL-Musaylh Mohanad S. , Shallal Qahtan Makki
Published: May 2022

Developed intelligent technologies are become play a promising role in providing better decision-making and improving the medical services provided to the patients. A risk prediction task for short-term is big challenge task; however, it is a great importance for recommendation systems in health care field to provide patients with accurate and reliable recommendations. In this work, clustering method and least square support vector machine are used for prediction a short-term disease risk prediction. The clustering similar method is based on euclidean Distance which used to identify the similar sliding windows. The proposed model is trained by using the slide windows samples. Finally, the appropriate recommendations are generated for heart diseases patients who need to take a medical test or not for following day using least square support vector machine. A real dataset which collected from heart diseases patient is used for evaluation. The proposed method yields a good results related by the recommendations accuracy generated to chronicle heart patients and reduce the risk of incorrect recommendations.
Pele negra, jalecos brancos: racismo, cor(po) e (est)ética no trabalho de campo antropológico
Authors: Castro Rosana
Published: May 2022

Durante pesquisa de campo de doutorado, acompanhei o trabalho de algumas médicas brancas em suas atividades de condução de protocolos de pesquisa clínica. Minha presença nos consultórios foi condicionada ao uso de um jaleco branco, peça que, por vezes, colocou-me em posição de explicar aos pacientes que não era uma estagiária de medicina e, por outras, tornou explícitos os limites de confusões supostamente automáticas entre mim e uma profissional da medicina. Por meio de uma análise de situações de racismo genderificado que vivi durante o trabalho de campo enquanto vestia um jaleco, reflito sobre o campo da medicina como espaço marcado pela branquidade e, estendendo tal crítica à antropologia, argumento que a reflexão ética sobre a pesquisa de campo deve levar em conta, necessariamente, as hierarquizações raciais e de gênero que compõem as interações com interlocutores de pesquisa – em especial, as experimentadas por pesquisadoras negras em contextos nos quais a branquidade é normalizada.
O tempo do mutirão: sentidos e (re)arranjos de família e casa entre mutirantes em um bairro periférico de São Paulo
Authors: Filadelfo Carlos
Published: May 2022

O Movimento dos Trabalhadores Sem Terra Leste I é um movimento social de luta por moradia que historicamente, desde a década de 1980, obtém atendimento para famílias militantes via mutirão autogestionário. O aqui denominado ‘tempo do mutirão’ refere-se a uma marcação temporal por parte dos mutirantes do período de duração do mutirão, desde o alcance da pontuação necessária e a decisão em de fato integrar uma demanda específica, passando por todo o processo de obra e chegando à mudança para a nova casa. Este artigo discute as inter-relações entre os termos família e casa a partir de narrativas de mutirantes da Leste I, sobre seus (re)arranjos familiares e domésticos durante e depois do mutirão a fim de realizar algumas reflexões etnográficas sobre as relações entre Estado e movimentos de moradia no atendimento habitacional a essas famílias.
Mitologías “populares”, elaboraciones eruditas
Authors: Sáez Oscar Calavia
Published: May 2022

Las mitologías “cultivadas” o “eruditas” de los pueblos amerindios han permanecido en general en el margen del análisis mitológico de tradición estructuralista. Comparando dos conjuntos de relatos históricos de origen -el uno mexica, tomado de cronistas indígenas del siglo XVI y el otro elaborado en la actualidad por autores del Alto Rio Negro, en la Amazonia- este artículo quiere mostrar la conexión, y sobre todo la dependencia, de este tipo de relatos con aquellos otros mitos que, en una ocasión, Lévi-Strauss llamó “populares”, y en los que basaba su propuesta analítica. Se señala el valor que las observaciones de Lévi-Strauss sobre la “muerte” o la “degradación” de los mitos tiene para entender las elaboraciones narrativas de los especialistas indígenas.
Abstract 106: Novel Nanoparticle-mediated Robust Genome Editing Targeting The Vascular Endothelium Of Postnatal And Adult Mice
Authors: Zhang Xianming , Huang Xiaojia , Jin Hua , Chaurasiya Birendra , Dong Daoyin , Zhao Youyang
Published: May 2022

Current viral and non-viral delivery of transgene after i.v. administration is mainly enriched in the liver. To target the vascular endothelium for robust genome editing in adults, we developed a a novel nanoparticle system. A single i.v. administration of the mixture of nanoparticles and all-in-one plasmid DNA expressing Cas9 controlled by
CDH5
promoter and guide RNA by
U6
promoter induced highly efficient genome editing in endothelial cells of the vasculatures including lung, heart, aorta, and the peripheral vessels in adult mice in as quick as 5 days. Western blotting and immunofluorescent staining demonstrated an 80% decrease of protein expression selectively in endothelial cells, resulting in a phenotype similar to that of genetic knockout mice. This nanoparticle /plasmid delivery system could also deliver a transgene targeting the vascular endothelium simultaneously and/or multiple gRNAs to knockout multiple genes at the same time. The nanoparticle/plasmid DNA delivery system could also achieve robust gene-correction through a base editor in the vascular endothelium in adult mice. These data demonstrate that nanoparticle delivery of plasmid DNA expressing a genome editing system (e.g., CRISPR/Cas9, or base editor, etc.) and/or a transgene is a simple powerful tool to rapidly and efficiently alter expression of gene(s) in vascular endothelium. This provides a significant advance in cardiovascular research and a potential novel gene therapy strategy for cardiovascular diseases.
Abstract 113: Cell-specific Chromatin Landscape Of Human Coronary Artery Resolves Mechanisms Of Disease Risk
Authors: Turner Adam W , Hu Sheng-en , Verdezoto Mosquera Jose E , Ma Wei Feng , Hodonsky Chani J , Wong Doris , Auguste Gaelle E , sol-church katia , Farber Emily , Kundu Soumya , Kundaje Anshul B , Lopez Nicolas G , Ma Lijiang , Ghosh Saikat , Onengut-Gumuscu Suna , Ashley Euan A , Quertermous Thomas , Finn Aloke , Leeper Nick J , Kovacic Jason C , Bjorkegren Johan L , Zang Chongzhi , Miller Clint L
Published: May 2022

Coronary artery disease (CAD) is a complex inflammatory disease involving genetic influences across several cell types. Genome-wide association studies (GWAS) have identified over 170 loci associated with CAD, where the majority of risk variants reside in noncoding DNA sequences impacting
cis
-regulatory elements (CREs). Here, we applied single-cell ATAC-seq to profile 28,316 cells across coronary artery segments from 41 patients with varying stages of CAD, which revealed 14 distinct cellular clusters. We mapped over 320,000 accessible sites across all cells, identified cell type-specific elements, transcription factors, and prioritized functional CAD risk variants via quantitative trait locus and sequence-based predictive modeling. Using differential peak analyses we identified a number of candidate mechanisms for smooth muscle cell transition states (e.g. fibromyocytes). By integrating these profiles with GWAS meta-analysis summary data we resolved cell type-specific putative binding sites for the majority of CAD risk variants. In particular, we prioritized functional variants predicted to alter MEF2 binding in smooth muscle cells at the MRAS locus. We also identify variants predicted to alter macrophage-specific regulation of LIPA. We further employed DNA to gene linkage to nominate disease-associated key driver transcription factors such as PRDM16 and TBX2. Together, this single cell atlas provides a critical step towards interpreting
cis
-regulatory mechanisms in the vessel wall across the continuum of CAD risk.
Abstract 116: Platelet-Induced Neutrophil Extracellular Traps Drive Ischemic Stroke Brain Injury
Authors: Denorme Frederik , Portier Irina , Cody Mark J , Neal Matthew D , Rondina Matthew T , Majersik Jennifer J , Yost Christian C , Campbell Robert A
Published: May 2022

Ischemic stroke prompts a strong inflammatory response which is associated with exacerbated stroke outcomes. However, classic anti-inflammatory strategies have been unsuccessful in stroke patients implying other unknown mechanisms contribute to injurious inflammation in stroke. Increasing evidence suggests immunothrombosis, a process involving coagulation, neutrophil and platelet activation, and neutrophil extracellular trap (NET) formation, is an important contributor to cardiovascular diseases. However, mechanistic regulators of immunothrombosis and their role in ischemic stroke remain unclear.

We examined markers of immunothrombosis in ischemic stroke patients and matched healthy donors. Stroke patients had significantly increased levels of D-Dimers, platelet factor 4, neutrophil calprotectin, citrullinated histone H3 (H3cit) and MPO-DNA complexes, markers of NET formation. In particular, H3cit and MPO-DNA complexes positively correlated with long-term stroke outcomes. Mechanistically, we observed increased plasma and platelet HMGB1 in stroke patients, which significantly correlated with plasma NETs, indicating a role for platelet HMGB1 in NET formation. To directly examine the role of platelet HMGB1, we employed a transient ischemic stroke mouse model. Depleting platelets significantly reduced plasma HMGB1 levels, inhibited NET formation and improved stroke outcomes. Correspondingly, administrating a HMGB1 inhibitor reduced NET formation and improved stroke outcomes, implying a causative role for platelet HMGB1 in mediating NET formation after stroke. As NETs appeared detrimental in ischemic stroke, we investigated the therapeutic potential of an endogenous NET inhibitory factor (NIF), recently discovered in neonates. Mice pretreated with NIF had reduced brain injury, improved neurological and motor function and enhanced survival after stroke. Importantly, NIF specifically blocked NET formation after stroke without affecting brain neutrophil recruitment. Critically, NIF still improved stroke outcomes when administered after stroke onset.

These results support a pathological role for NETs in stroke brain injury and indicate the use of NIF as a therapeutic strategy to target immunothrombosis in stroke.
Abstract 123: Intestinal Deletion Of 3-hydroxy-3-methylglutaryl-coenzyme A Reductase Promotes Expansion Of The Resident Stem Cell Compartment
Authors: Doerfler Alexandria M , Han Jun , Tang Li , De Giorgi Marco , Jarrett Kelsey E , Hurley Ayrea , Li Ang , Chuecos Marcel , Morand Pauline , Ayala Claudia , Martin James F , Goodlett David R , Vallim Thomas A , Shroyer Noah , Lagor William R
Published: May 2022

While the intestine is the critical interface between cholesterol absorption and excretion, surprisingly little is known about the role of
de novo
cholesterol synthesis in this organ and how it affects whole body cholesterol homeostasis. The mevalonate pathway is most well-known for the production of cholesterol, but it is also required for the production of essential non-sterol isoprenoids. 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), the rate-limiting enzyme in the mevalonate pathway, is regulated by a three-protein complex made up of INSIG, SCAP, and SREBP2. Intestine specific knockouts of
Scap
and
Srebp2
both result in severe enteropathy and reduced mouse survival. Here, we assessed the hypothesis that
Hmgcr
is required for enterocyte viability. Mice harboring floxed alleles for
Hmgcr
were bred with the
Villin-Cre
transgene to specifically knock out this enzyme in the villus and crypt epithelial cells of the small intestine (i-KO). The i-KO mice are viable through adulthood and fertile.
Hmgcr
is efficiently deleted based on analysis via drop digital PCR and qPCR. RNA sequencing shows reduction in all SREBP2 target genes throughout the mevalonate pathway in intestinal epithelial cells. Lipidomics confirms substantial reductions in abundance of all sterol and non-sterol isoprenoids, except 7-dehydrocholesterol and cholesterol. Cholesterol is likely maintained through reabsorption of biliary cholesterol or increased uptake from the circulation. Circulating cholesterol levels and cholesterol absorption are not altered in i-KO mice, but triglyceride absorption is increased through compensatory changes in bile acid composition and intestinal growth. At the cellular level, the intestine rapidly compensates for loss of
Hmgcr
via dramatic expansion of the stem cell compartment within the crypts. In conclusion, the mechanisms by which the intestine compensates for genetic loss of
Hmgcr
include altered triglyceride absorption, bile acid composition, increased absorptive surface area, and expansion of the resident stem cell compartment. Together these studies provide insight into the effects of HMGCR knockout in intestinal development and lipid metabolism.
Abstract 127: Aspirin Resistant Biomechanical Platelet Activation In Fibromuscular Dysplasia: Novel Mechanisms Of Platelet Activation & Stroke
Authors: Bhandari Rohan , Aggarwal Anu , Godwin Matthew , Pascual Crystal , Shim Sharon , Fendrikova-Mahlay Natalia , Hazen Stanley L , Elbadawi Ayman , Aslan Joseph E , CAMERON Scott J
Published: May 2022

Background:
Fibromuscular dysplasia (FMD) is a non-atherosclerotic, non-inflammatory disorder characterized by abnormal arterial morphology and turbulent blood flow. In regions of disturbed blood flow, circulating platelets may become activated. Cerebrovascular FMD is common and increases the risk of stroke in affected patients. Most FMD patients are treated with antiplatelet agents, however these medications target biochemical and not biomechanical pathways of platelet activation.

Methods:
Thrombotic outcomes were determined by multivariate regression analysis of 105,887 patients with FMD. Platelets were isolated from humans and activation was assessed by FACS and aggregometry following exposure to
ex vivo
steady laminar and disturbed flow (SF and DF, respectively) conditions using a cone and flow system. Single-cell phenotyping of control and FMD platelets with transcriptomics (RNAseq) and proteomics were performed. Cellular bioenergetics were evaluated by real-time metabolic analyses.

Results:
In patients with FMD, long-term aspirin therapy is paradoxically an independent risk factor for acute ischemic stroke (OR 1.64, 95% CI 1.29-1.94) but seems to protect against acute hemorrhagic stroke (OR 0.47, 95% CI 0.24-0.89). FMD platelet activation and aggregation were hyporeactive to biochemical agonists, but hyperreactive to
ex vivo
mechanical, disturbed blood flow conditions compared to age/gender-matched controls. FMD platelet activation in DF was attenuated by the mechanosensitive channel inhibitor, GsMTx4. Principle component analyses of RNAseq and proteomics identified markedly different platelet phenotypes with key differences in mitochondrial function/fission. Platelet imaging and metabolic analyses revealed abnormal mitochondrial architecture and respiration in FMD.

Conclusions:
FMD platelets display a divergent, dysregulated platelet phenotype with attenuated biochemical but augmented biomechanical activation. DF exposure of platelets in irregularly shaped arteries in FMD may alter the platelet phenotype and metabolic function. Therapeutics targeting biomechanical pathways of platelet activation may provide a superior mechanism for stroke prevention without unintended bleeding consequences.
Abstract 129: BAF60c Prevents Abdominal Aortic Aneurysm Formation Through Maintaining Vascular Smooth Muscle Cell Homeostasis
Authors: Zhao Guizhen , Zhang Jifeng , Chen Eugene , Zhao Yang , Lu Haocheng , zhu tianqing , Chang Ziyi
Published: May 2022

Background:
Abdominal aortic aneurysm (AAA) is a common and life-threatening vascular disease. Vascular smooth muscle cell (VSMC) dysfunction is involved in the dilatation and eventual rupture of AAA, with the precise epigenetic control of this process less studied. BAF60c, a unique subunit of SWItch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex, serves as a core modulator of physiological and pathological processes, yet little is known about its function in the vasculature and pathogenesis of AAA.

Methods and Results:
BAF60c is downregulated in AAAs from humans and mice with primary staining to the aortic VSMC, confirmed by single cell RNA sequencing. In vivo studies revealed that VSMC-specific knockout Baf60c markedly aggravated both AngII- and elastase-induced murine AAA formation, with significant increases of elastin degradation, inflammatory cell accumulation, VSMC phenotype switching, and apoptosis. In vitro studies showed that knockdown of BAF60c in human aortic smooth muscle cells resulted in the loss of contractile phenotype while upregulation of inflammatory genes and increased apoptosis. Chromatin immunoprecipitation followed by sequencing and qPCR, as well as CoIP assays showed that BAF60c maintained VSMC contractile phenotype by enhancing serum response factor association with its co-activator p300 and SWI/SNF complex, suppressed VSMC inflammation via precise control of the repressive state of NF-κB-target genes, as well as prevented VSMC apoptosis through transcriptional activation of KLF5-dependent BCL2 expression.

Conclusions:
Our study indicates that BAF60c is essential for SWI/SNF complex recruitment to chromatin and interaction with diverse transcription factors to precise epigenetic control of VSMC homeostasis and prevention of AAA formation.
Abstract 130: Wnt16 Deficiency Worsens Angiotensin Ii - Induced Aneurysmal Remodeling Of Ascending Thoracic Aorta
Authors: Behrmann Abraham S , Zhong Dalian , Li Li , Sabaeifard Parastoo , Goodarzi Mohammad , Lemoff Andrew , Towler Dwight A
Published: May 2022

Wnt16 controls bone strength via paracrine osteoblast-osteoclast remodeling interactions.
Wnt16
mRNA is highly expressed in both aorta & bone, & immunofluorescence (IF) localized Wnt16 to vascular smooth muscle (VSM). Paracrine VSM - monocyte/macrophage (M/MAC) interactions control arterial remodeling, resembling osteoblast-osteoclast interactions in bone. We assessed the impact of Wnt16 deficiency on aneurysmal remodeling (6-7/group). Angiotensin II infusion for 4 weeks increased ascending thoracic aorta diameter by 60% (Holm-Sidak p = 1.5E-4, ANOVAp = 2.7E-6) in male LDLR-/- mice. Wnt16-/-;LDLR-/- cohorts exhibited worsened ascending aorta dilatation (by 20%; Holm-Sidak p = 0.03), indicating a protective role for Wnt16. Mass spectrometry of thoracic aorta proteins from Wnt16-/-;LDLR-/- vs. LDLR-/-mice revealed a 9-fold increase in MMP12 (p = 0.008; n=3/group) — a M/MAC protease key to aneurysmal remodeling. IF confirmed enhanced adventitial MMP12 in aortas of Wnt16-null mice. Treatment of M/MACs with Wnt16 (8 nM) down-regulated
Mmp12
mRNA (78%; p < 0.001), with increased
IL10
(2-fold; p < 0.05) &
Arg1
(6-fold; p =0.001). Hypomorphic alleles encoding contractile proteins predispose to hereditary thoracic aortic aneurysm; thus, we studied the impact of Wnt16 deficiency on the contractile phenotype. VSM from Wnt16-null thoracic aorta exhibited reductions in
Myh11
(down 60%; p < 0.05), Acta2 (by 35%, p <0.05), & the VSM transcriptional co-activator
Myocardin (
by 65%; p = 0.0002), while
Axin2
was increased 1.7-fold (p = 0.01). Acute siRNA-mediated knockdown of
Wnt16
in VSM -M/MAC admixtures (95% VSM, 5% M/MAC) also downregulated contractile genes & upregulated
Mmp12
. This was phenocopied by siRNA targeting
Taz/Wwtr1 —
a transcriptional collaborator of Myocd we identified as exhibiting impaired nuclear localization in Wnt16-null VSM.
In vivo
dosing with Wnt16 (0.25 mg/kg/day, 2 days) upregulated aortic contractile gene expression 1.6- to 1.8-fold, with
Mmp12
reduced 50% (p < 0.05), as consistent with the reciprocal changes upon Wnt16 deficiency. Wnt16 limits thoracic aortic remodeling, preserves the VSM contractile phenotype, & reduces Mmp12. Strategies that augment Wnt16 signaling may mitigate enlargement of thoracic aortic aneurysms.
Abstract 131:
Gata4
Ablation In Smooth Muscle Cells Reduces Aortic Root Dilation In A Murine Model Of Loeys-Dietz Syndrome
Authors: Bramel Emily E , Saqib Muzna , Creamer Tyler J , Gallo MacFarlane Elena
Published: May 2022

Loeys-Dietz syndrome (LDS) is a connective tissue disorder characterized by predisposition to aneurysm. Although dilation can develop in all segments of the arterial tree, the aortic root is especially prone to disease. This aortic region is primarily composed of vascular smooth muscle cells (VSMCs) derived from secondary heart field (SHF) progenitors, while VSMCs derived from the cardiac neural crest (CNC) predominate in the more distal ascending aorta. Although intrinsic differences have been found between these two types of VSMCs, it remains unclear how these may contribute to localized risk. To identify transcripts associated with the vulnerability of the aortic root in LDS, we performed single cell transcriptomic analysis on proximal aortas from 16-week-old control and

Tgfbr1
M318R/+

LDS mice, which carry a kinase-inactivating mutation in TGF-β Receptor 1. We identified four major VSMC clusters regardless of genotype, with two clusters being defined by CNC-enriched transcripts and two clusters by SHF-enriched transcripts. The latter were defined by expression of
Gata4
, which codes for a transcription factor required for development of SHF-derived vascular structures, as well as by higher expression of transcripts previously involved in aneurysm pathogenesis such as
Thbs1
,
Tgfb1,
and
Tgfb3.
In situ RNA hybridization confirmed that
Gata4
-expressing VSMCs were enriched in the aortic root and proximal ascending aorta, with almost no expression in the more distal ascending aorta, suggesting that this transcript defined the aortic region most susceptible to the effects of the

Tgfbr1
M318R/+

mutation. GATA4 protein levels were also upregulated in the aortic root of LDS mice. To test if excessive GATA4 expression or activity in VSMCs contributed to aortic root dilation in LDS, we generated LDS mice in which
Gata4
was specifically deleted in VSMCs by tamoxifen administration at 6 weeks of age. We found that this intervention significantly reduced aortic root growth from 8 to 20 weeks of age. Although the mechanisms remain under investigation, these data suggest that expression of
Gata4
might sensitize specific subsets of aortic VSMCs to the effects of an LDS mutation, rendering the aortic root more susceptible to disease.
Abstract MP05: Sex Differences In Mortality And 30-day All Cause Readmission Following Carotid Endarterectomy Following Acute Ischemic Stroke: A Nsqip Study (2014 - 2017)
Authors: Suarez Mariana M , Saxena Anshul , Ramamoorthy Venkataraghavan , Rubens Muni , McGranaghan Peter , Veledar Emir , Garelnabi Mahdi O
Published: May 2022

Background:
Studies report that acute ischemic stroke (AIS) affects males and females differently. For example, the treatment outcomes of intra-arterial thrombolysis differ between males and females. In this study, we examined mortality and 30-day readmission differences by sex among AIS patients who had carotid endarterectomy (CEA).

Methods:
We used data from National Surgical Quality Improvement Program (NSQIP) registry (2014-2017). Patients ≥18 years of age, with CEA for AIS were included. AIS and CEA were identified using ICD-9 and ICD-10 diagnosis, and CPT codes, respectively. Using machine learning methods such as Hierarchical clustering, we grouped patients (low, medium, and high-risk clusters) based on their demographics, past medical history, and preoperative variables. Differences in means, and differences in proportions were calculated. Logistic regression was conducted for 30-day readmission and survival analysis for mortality, accounting for cluster groups and sex.

Results:
There were a total of 22,373 AIS patients who received CEA treatment. Mean (SD) age of the sample was 70.7 (9.4) years, and 61% were males while 39% were females. Mortality rates were 0.8% and 0.7% for men and women (p=0.113), respectively. Thirty-day readmission rates were 6.3% and 7.6% for men and women (p<0.0001), respectively. There were 56.3%, 34.4%, and 9.3% patients in Low, Medium, and High-Risk clusters, respectively. Females were 1.2 times (OR 95% CI: 1.1 - 1.3) as likely to be readmitted compared to males. Survival analysis showed that there was no significant difference in mortality between males and females (HR: 0.8; 95% CI: 0.6 - 1.2; p = 0.28).

Conclusion:
Our study found sex related disparities in short-term readmissions as it was higher among females. This could be because of underlaying sex specific pathophysiology of AIS. Healthcare providers should consider sex-specific management to improve post-stroke recovery for women and reduce their excess burden.
Abstract MP08: Telomerase Reverse Transcriptase Mediates Osteogenesis In Calcific Aortic Valve Disease
Authors: Cuevas Rolando A , Hortells Luis , Boufford Camille , Regan Cailyn , Wong Ryan , Crane Alexander , Chu Claire , Moorhead William , Lee Angela , Bashline Michael , Gurkar Aditi , Bruemmer Dennis , rojas mauricio , Gleason Thomas G , Billaud Marie , Sultan Ibrahim , St Hilaire Cynthia
Published: May 2022

Calcific aortic valve disease (CAVD) is the leading heart valve disorder in the US. It is characterized by an active accumulation of calcium nodules on the aortic valve leaflets which lead to stiffening and remodeling of the valve leaflets causing valve dysfunction, cardiac failure and increased stroke risk. Inflammation and mechanical stresses contribute to CAVD pathogenesis. However, the mechanisms driving the fibrocalcific remodeling of the aortic valve are currently ill-defined. Multiple studies have revealed that the catalytic subunit of telomerase reverse transcriptase (TERT) can induce gene transcription and its overexpression primes mesenchymal stem cells to differentiate into osteoblasts, suggesting that TERT has a role in the activation of osteogenic transcriptional programs. We hypothesized that TERT contributes to early events leading to calcification of the valve leaflet. In human calcified valve tissue, we found that TERT protein is highly expressed in areas of calcification compared to control valve tissue, with no effect on telomere length. Alpha-SMA, a VIC activation marker, and RUNX2, a key transcription factor involved in the osteogenic differentiation of osteoblast, were also elevated in CAVD tissue. Under osteogenic differentiation conditions, human valve interstitial cells (VICs) upregulated TERT, RUNX2, and alpha-SMA protein levels and calcified, while CAVD VICs calcified de novo. Inflammatory stimuli intensified in vitro calcification, and induced TERT, RUNX2, and alpha-SMA protein expression. PLA and ChIP analysis showed that TERT interacts with interacted with Signal Transducer and Activator of Transcription 5A/B (STAT5) and together bind to
RUNX2
promoter, respectively. shRNA-mediated TERT downregulation reduced expression of RUNX2 and alpha-SMA and genetic deletion of
Tert
in murine mesenchymal stem cells and vascular smooth muscle cells prevented calcification. These data provide evidence that TERT is required for calcification, regulates the transition of quiescent VICs into calcifying VICs, and that STAT5 functions as a TERT-interacting partner for DNA binding.
Abstract MP09: MLKL (Mixed Lineage Kinase Domain-like Protein) Is A Novel Regulator Of The Splenic Microenvironment That Restricts Hematopoiesis During Atherosclerosis
Authors: Rasheed Adil , Wyatt Hailey , Dennison Taylor , Nguyen My-anh , Robichaud Sabrina , Geoffrion Michele , Baxi Adir , Lee Richard , Ouimet Mireille I , Rayner Katey J
Published: May 2022

During atherosclerosis, macrophages within the aorta undergo necroptosis, a mode of pro-inflammatory cell death that is triggered by the phosphorylation of RIPK1 & RIPK3 and leads to the activation of the mixed lineage kinase domain-like protein (MLKL) and cell lysis. Our previous work demonstrated that while inhibition of MLKL decreased necroptosis and necrotic core in the plaque, unlike RIPK1 & RIPK3 this inhibition did not lead to overall decrease in plaque area, suggesting that MLKL may in fact participate in other processes that drive atherosclerotic plaque development. Because circulating leukocytes are recruited and contribute to the growing atheroma, and these leukocytes are derived from hematopoietic stem and progenitor cells (HSPCs) found in both the bone marrow (BM) and the spleen, we investigated whether these hematopoietic reservoirs became dysregulated upon loss of MLKL during atherogenesis. To induce atherogenesis,
Apoe
-knockout mice (
Apoe
-/-) were fed a high fat-high cholesterol diet for 16 weeks while receiving weekly subcutaneous administration of antisense oligonucleotides (ASOs) to knockdown MLKL expression. In the
Apoe
-/- mice receiving MLKL ASO we observed a 3.1-fold increase in splenomegaly compared to the control ASO, specifically with an expansion of the splenic red pulp, the area of splenic hematopoiesis. Furthermore, flow cytometry revealed a significant increase in myeloid HSPCs and mature myeloid populations in the spleen, but not the BM, after MLKL knockdown. However, no changes were observed in these hematopoietic measures after transplantation of
Ldlr
-knockout mice with MLKL-knockout BM, indicating that MLKL restricts hematopoiesis through its regulation of non-hematopoietic cells of the splenic niche. Indeed, upon MLKL knockdown in the
Apoe
-/- mice we observed a >50% reduction in splenic endothelial cells, which are required to promote HSPC quiescence and repress hematopoiesis. These data therefore demonstrate a novel role for MLKL on preserving splenic endothelial cells to limit immune cell development during atherosclerosis, and more generally, highlight the importance of the dysregulation of the splenic microenvironment to contributing to the inflammatory processes that drive atherosclerosis.
Abstract MP17: TLR4 May Play A Role In Accelerated Atherosclerosis In Beta Thalassemia
Authors: Hurtado Julian , Sellak Hassan , Archer David R , Taylor W. Robert R
Published: May 2022

Beta Thalassemia (BT) is an autosomal recessive hemoglobinopathy that affects 80 million people worldwide. Children with BT display an increase in carotid-intima media thickness, an early sign suggestive of premature atherosclerosis. However, it is unknown if there is a direct relationship between BT and atherosclerotic disease. BT has increased free heme leading to ROS generation directly or via Toll Like Receptor (TLR4) activation, but the role of heme in disease progression in BT is poorly characterized. The aim of this study is to evaluate the progression of atherosclerosis in BT mice and the potential role of TLR4. Atherosclerosis was induced by implementing short (ST) and long term (LT) models. In our ST model, both BT (

Hbb
th-3

) and wild type (b6) male/female littermates (n=5) were fed a Paigen high fat diet (PD) for 8 weeks. On week 2, mice were injected with AAV8-PCSK9 to knockdown the LDL receptor. On weeks 4-8, mice were treated with Angiotensin ll via subcutaneous osmotic pumps implantation. The LT model was performed by placing mice on 3 months of PD with PCSK9 overexpression. Atherosclerosis was evaluated by atherosclerotic plaque lesion area of the descending aorta via
en face
analysis, histology of the aortic roots, and RT-qPCR of inflammatory markers. In addition, to evaluate the possible contribution of TLR4 in atherosclerosis, bone marrow transplants were performed by grafting WT and BT bone marrow (n=2-4) to both b6 and TLR4
-/-
mice, and atherosclerosis was performed as described in our LT model.In our ST model, aortic
en face
analysis revealed elevated plaque accumulation in BT mice (WT: 22 ± 4% plaque area vs. Thal: 45 ± 11% plaque area, p = .024). Aortic root (H&E) analysis did not display a significant difference in our ST model, but it was significantly increased in the LT model. Gene expression profiling indicated a significant increase in both IL-6 and CD-68 (p<0.041) in BT mice. In our BMT experiments, we saw a significant decrease in aortic root plaque in TLR4
-/-
mice with Thal BM (.24 ± .06 mm
2
) compared to wild type mice with Thal BM (.39 ± .07 mm
2
). Our data demonstrate for the first time that the underlying pathophysiology of BT clearly leads to accelerated atherosclerosis and suggest TLR4 is playing a role in atherosclerotic development in BT.
Abstract MP26: Activating P2Y
1
Receptors Improves Function In Arteries With Repressed Autophagy
Authors: Cho Jae , Park Seul-Ki , Kwon Oh Sung , La Salle D. Taylor , Cerbie James , Fermoyle Caitlin C. , Morgan David , Nelson Ashley , Bledsoe Amber , P Bharath Leena , Tandar Megan , Kunapuli Satya P , Richardson Russ , Anandh Babu Pon Velayutham , Mookherjee Sohom , Kishore Bellamkonda K , Wang Fei , Yang Tianxin , Boudina Sihem , Trinity Joel D , Symons J David D
Published: May 2022

Objective:
The importance of endothelial cell (EC) autophagy to vascular homeostasis is evolving. Earlier we reported that purinergic 2Y
1
receptor (P2Y
1
-R) activation rejuvenates shear-stress induced nitric oxide (NO) generation in bovine aortic endothelial cells that is otherwise compromised after pharmacological and genetic autophagy repression. Here we determined the translational and functional relevance of these findings.

Approach and Results:
First we assessed translational relevance using older humans and mice that exhibit blunted EC autophagy at rest together with impaired arterial function vs. appropriate controls. Rhythmic handgrip exercise elevated radial artery shear rate similarly in adult and older males for 60-min. Compared to baseline, autophagy initiation, p-eNOS
S1177
activation, and NO generation, occurred in radial artery ECs from adult but not older subjects.Regarding mice, indexes of autophagy and p-eNOS
S1177
activation were robust in ECs from adult but not older mice in response to 60-min treadmill-running. Next we questioned whether an inability to initiate EC autophagy precipitates arterial dysfunction. Age-associated reductions in intraluminal flow-mediated vasodilation observed in older vs. adult mice were recapitulated in arteries from adult mice by : (i) NO synthase inhibition; (ii) autophagy impairment using 3-methyladenine (3-MA); (iii) EC Atg3 depletion (iecAtg3KO mice); (iv) P2Y
1
-R blockade; and (v) germline depletion of P2Y
1
-Rs. Importantly, P2Y
1
-R activation using 2-methylthio-ADP (2-Me-ADP) improved vasodilatory capacity in arteries from : (i) adult mice treated with 3-MA; (ii) adult iecAtg3KO mice; and (iii) older animals with repressed EC autophagy.

Conclusions:
Arterial dysfunction concurrent with pharmacological, genetic, and age-associated EC autophagy compromise is improved by activating P2Y
1
-Rs.
Abstract MP29: Skeletal Muscle Satellite Cells Enhance Vascular Growth Through Paracrine Signaling
Authors: Joseph Giji , Yu Tao , Mavris Sophia , Taylor W. Robert R , Hansen Laura M
Published: May 2022

Peripheral arterial disease (PAD) is a major health problem that affects over 200 million people worldwide. Decreased blood flow to the limb muscles leads to ischemia that results in pain, decreased quality of life, and in severe cases amputation. Despite the prevalence and severity of the disease, effective treatment options are still limited. One factor correlated with improved prognosis is the generation of a more robust collateral vessel network. This study hypothesized that skeletal muscle satellite cells, which play a key role in skeletal muscle regeneration, also contribute to vascular regeneration in the setting of ischemia. Specifically, satellite cells are proposed to generate cytokine and growth factors which modulate vascular growth via paracrine signaling. Satellite cells were isolated and cultured for vasculogenic assays include migration co-cultures. Satellite cells encapsulated in alginate were delivered to a hindlimb ischemia model of vascular growth to assess their therapeutic potential in vivo. Gene expression of satellite cells from ischemic tissue was assessed using a microarray. The migration assays demonstrated that satellite cells produce chemokines which increased smooth muscle migration (3.5 fold) and endothelial cell migration (2.8 fold) over control conditions (n = 4, p<0.05) In the hind limb ischemia model, alginate encapsulated satellite cells increased perfusion 17% closer to baseline (68% vs 51%, n = 11, p<0.05) measured via Laser Doppler imaging. Capillary density as measured by Lectin staining increased 1.6 fold and smooth muscle positive vessels increase more than 2 fold (n=6, p<0.05). Finally, Ingenuity pathway analysis of the gene array day suggested that vasculogenic and cell migration pathways were increased. Taken together, these findings demonstrate that satellite cells produce a number of factors that can serve as chemokines in vitro and increase vascular growth in vivo. Further work will explore the mechanisms by which satellite cells exhibit their effects and develop a therapeutic application of satellite cells as a novel treatment for patients with peripheral artery disease.
Abstract MP32: Diaphanous 1 Regulates Actin Polymerization In Adipocytes: Potential Mechanism Regulating Obesity In Mice
Authors: Ruiz Henry H , Schmidt Ann Marie
Published: May 2022

The underlying pathophysiological mechanisms of obesity, a hallmark of metabolic and cardiovascular diseases, are not fully elucidated. Adipocyte hypertrophy contributes to insulin resistance, and is coupled to meaningful alterations in actin dynamics that favor actin polymerization. Diaphanous 1 (Diaph1) is a formin protein that associates with the fast-growing/barded end of actin filaments to regulate actin dynamics. We hypothesized that deletion of
Diaph1
or pharmacological regulation of actin polymerization in the context of lipid surplus would prevent obesity, decrease adipocyte lipid load and protect from obesity-associated metabolic disease. We modeled obesity by feeding wild type (Wt) mice and mice globally or with adipocyte-specific deletion of
Diaph1
a high fat diet (60%kcal from fat). Body composition was assessed by Dual-energy X-ray absorptiometry (DEXA) and dynamic glucose and insulin tolerance testing was performed to assess metabolic state.
In-vitro
studies were carried out in adipocytes differentiated from primary pre-adipocytes or 3T3-L1 fibroblasts. Lipid overload was modeled using a mixture of palmitate/oleate (P/O). Actin manipulations were achieved by pharmacologically preventing (Latrunculin B [LatB]) or promoting (Jasplakinolide [Jasp]) F-actin polymerization. Mice globally devoid of, or with adipocyte specific
Diaph1
deletion were protected from obesity, glucose intolerance and insulin resistance. Wt adipocytes treated with P/O exhibited pronounced accumulation of triglycerides (Tg) which was absent in adipocytes differentiated from
Diaph1
knock out mice. Pharmacological studies in 3T3-L1-derived adipocytes confirmed the P/O-induced Tg accumulation which was prevented by LatB and exacerbated by Jasp. We present the novel finding that adipocyte DIAPH1 is a major player in the pathophysiology of obesity and associated metabolic sequela which results, at least in part, from regulation of adipocyte hypertrophy by preventing actin polymerization. Thus, we propose that actin remodeling is an underlying mechanism for adipocyte hypertrophy in obesity and inhibition of DIAPH1 may represent a potential therapeutic target for the treatment of obesity and related metabolic and cardiovascular disorders.