bluuebunny/crossref_metadata_2025 huggingface dataset:
Urban Uses Put to The Test by The Covid-19 Pandemic: A Literature Review and Evidence from Algeria
Authors: Naimi Ait-Aoudia Meriem , Khettab Samira , Boukratem Oumelkheir
Published: July 2024

The COVID-19 pandemic advent has put the spotlight back on debates about the relationship between urban planning and health-related issues, prompting a certain amount of research works worldwide. This study set out to build an analysis grid for this relationship on the basis of a literature review and apply it to the case study. To this end, our work used two levels of bibliographical analysis: the first level focused on review of research papers, which helped identify three topics related to the consequences of coronavirus disease and subsequent restrictive measures on urban uses, namely public open space, mobility, and online uses. Our study considers these latter because of their impacts on urban use and, by ricochet, on urban planning and design. The next level based exclusively on original articles review, allowed more in-depth investigations, to break down these topics into sub-topics, and to define criteria for analyzing them, which mainly include use patterns and behavioral changes and perceptions. Drawing from the insights revealed by these studies, the present article endeavors to examine the Algerian case with the aim of nourishing the debate on the resilience of cities to health crises and suggesting better guidance for urban planning and design measures.
Psychological Social Behavior: Taxable Entrepreneurs Respond to Vat Policy Changes
Authors: Wibowo Danny
Published: July 2024

Environmental considerations or individual intentions influence taxpayer behavior. VAT changes may be a reaction to behavior either overtly or covertly. This research applies an approach approach with the method of interviews conducted with business actors. However, VAT can reduce profits by influencing consumers to spend resources and look for lower prices. Regarding the VAT rate, it is still considered unfair. Furthermore, double recording or splitting the business into several forms is implemented, which is passed on to other family members. Another option is to do tax planning to meet the tax burden. Decisions in the information action model can be based on rationality, routine behavior, nature/tradition and subjective norms / perceptual behavior. Findings from an environmental perspective are formed from the level of experience and age, as well as the encouragement of social communities and provide psychological stimuli for taxpayer behavior in assessing changes in tax regulations, especially about VAT rates.
Flood Risk Assessment for Better City Resilience; Case of The Bab El Oued Agglomeration, Algiers (Algeria)
Authors: Taib Lyes
Published: July 2024

The concept of natural risk represents a challenge to the economic and social development of a given territory, which leads to the adoption of a rational and studied approach to addressing the critical situation imposed by risks. Flooding is one of the most dangerous phenomena, capable of causing enormous damage to life, the economy, and the environment.
The locality of Bab el Oued experienced a painful episode in its history in terms of flood damage during the year 2001. The risk in this locality is classified as very high, which implies that it is necessary to continuously reassess preventive measures and disaster management strategies if flooding occurs. This should be based on both natural and human factors, making risk a devastating reality and a constraint on sustainable development.
Through this article, attempts have been made to find intelligent solutions for flood disaster management in the highly threatened neighbourhood of Bab El Oued within the framework of current legislation and regulations, following a likely scenario.
The Role of Transformational Leadership in Managing Human Resources for Organizational Innovation Case Study in A State Electricity Company
Authors: Amir  , Adiwijaya  , Firly Juanita Surahman  , Syam Mukhlisah
Published: July 2024

Transformational leaders provide autonomy and responsibility to employees, allowing them to take initiative and contribute actively to organizational goals. The aim of this research is to understand the role of transformational leadership in influencing human resource management strategies to support innovation within the State Electricity Company. The research method used in this study is Systematic Literature Review (SLR). The results of this study indicate that transformational leadership is not just about managing organizations but also about building a culture that continuously seeks ways to improve and innovate, ensuring that the State Electricity Company remains competitive and relevant in facing the challenges of the future electricity industry.
Morphological, physiological and anatomical traits in durum wheat (Triticum durum Desf.) as affected by semi-arid conditions
Authors: Ali Arous , Zemour Kamel , Mehdeb Djamila , Labdelli Amina , Zemour Hafidh , Chouhim Kada Mohamed Amine , Adda Ahmed , Belkhoudja Moulay
Published: July 2024

Durum wheat (Triticum durum Desf.) is one of Algeria's primary cereal crops. Although the areas reserved for this species are estimated at approximately 40%, the production remains low to meet national grain requirements. This shortfall is primarily due to the impact of biotic and abiotic stresses, particularly drought, which significantly limit wheat yields.  For that, this study assessed the behavior of five genotypes of durum wheat (Waha, Acsad1361, Vitron, Oued Zenati and Langlois) under water stress conditions. The experiment was carried out in Tiaret (Western Algeria). The studied parameters are morpho-physiological and anatomical of the flag leaf. The obtained results showed that the water deficit greatly affected the dimensions of the plant and triggered a decrease in the water content. Structural modifications due to cellular resizing of the structural constituents of the last leaf, reflected by variations in the rate of the outer epidermal wall thickness and reductions of leaf mesophiles. These modifications remain effective in maintaining the hydration of the aerial part by limiting water loss and increasing the hydraulic resistance of the leaves. According to this study, it seems that an inter-varietal difference has been highlighted. However, a genetic cross is recommended to combine the maximum number of resistant genes into a single variety. This strategy is considered as an effective solution for mitigating the effects of abiotic stresses, especially in semi-arid regions.
Exploring Celestial Object Characteristics: An In-depth Analysis of Quasars, Stars, and White Dwarfs Using the Sloan Digital Sky Survey (SDSS) Dataset
Authors: Siagian Ruben Cornelius , Nurahman Arip , Sinaga Goldberd Harmuda Duva , Ariefka Reza , Pribadi Pandu
Published: July 2024

This research utilizes the Sloan Digital Sky Survey (SDSS) dataset, examining 12,884 observations to explore quasars, stars, and white dwarf objects. Magnitude data and coordinates across five filter bands are analyzed, revealing unique features through statistical methods. The identification of 77,429 quasars with 15 dimensions enhances the dataset. Thorough analyses of stellar and white dwarf classes, coupled with visualization techniques, unveil variable relationships. Residual validation and Gaussian kernel density plots confirm significant class differences. Non-linear regression and a normal distribution mixture model depict complex variable relationships. A parallel coordinates plot aids in interpreting data patterns, while predictive modeling via regression exposes meaningful coefficients. Logistic regression effectively classifies astronomical objects in the SDSS training data. This research contributes to understanding celestial object characteristics, offering valuable insights for astronomers and astrophysicists in analyzing large-scale astronomical datasets.
Abstract 1000: Platelets Act As Circulating Reservoir Of Tgfβ1: Role In Abdominal Aortic Aneurysms
Authors: Pratama Muhammad Yogi , Der Nataley , Rivera Cristobal , Sridharan Kousik , Giardinetto Catarina Santos , Silvestro Michele , Maldonado Thomas , Rhamkhelawon Bhama
Published: July 2024

Introduction:
Transforming Growth Factor-Beta1 (TGFβ1) signaling plays a crucial role in the development and maintenance of the vasculature by orchestrating vascular remodeling signaling cascades. Aberrant TGFβ1 leads to aortic aneurysms, yet there is a persistent conflict in its role in driving or protecting against the degradation of the extracellular matrix (ECM) in abdominal aortic aneurysms (AAA). Notably, limited studies have delved into its cell-specific actions. This ambiguity highlights the need for an improved understanding of the TGFβ1 cell-specific role in AAA.

Aims:
To elucidate the role of platelet-derived TGFβ1 in AAA development thereon expanding our knowledge on the role of TGFβ1 in vascular remodeling.

Methods and Results:
Single-cell epitope and transcriptome analysis by using cellular-indexing of transcriptomes and epitopes by sequencing (CITE-seq) of the blood of patients with AAA revealed that platelets were enriched with TGFβ1 compared to healthy individuals. Comparative pathway analysis of the platelet populations showed the association of this cluster with ROS, platelet activation and chemokine signaling pathways that promote AAA pathogenesis. These results were validated in the Apolipoprotein E deficient (ApoE
-/-
) mice treated with Angiotensin II for 28 days. To delve into the direct role of platelet-derived TGFβ1 in AAA, we generated mice with conditional knock-down of TGFβ1 in platelets (Tgfb1
flox/flox
Pf4
Cre+
) and subjected these mice to AAA. Remarkably, these mice exhibited reduced aneurysm development and preserved elastin microstructure compared to controls. The expression of phosphorylated smad2/3, one of the downstream TGFβ1-canonical signaling pathways, as well as matrix metalloproteinase, were repressed in the aorta of Tgfb1
flox/flox
Pf4
Cre+
mice, suggesting the platelets could supplement Tgfb1 to the aortic tissue sufficient to fuel matrix degradation. These results were recapitulated when the platelets were depleted from the circulation using an anti-GP1b antibody.

Conclusion:
Our findings reveal that platelets represent an important reservoir of Tgfb1, clarify its cell-specific role in ECM remodeling and underscore its detrimental role in sustaining matrix degradation during AAA.
Abstract 1001: Interleukin-33 Deficiency Exacerbated Angiotensin II-induced Abdominal Aortic Aneurysms
Authors: Asakawa Tomohiko , Umebayashi Ryoko , Sakurabu Yoshimasa , Katayama Katsuyoshi , Okamoto Shugo , Onishi Yasuhiro , Matsuoka-Uchiyama Natsumi , Takeuchi Hidemi , Uchida Haruhito A , Wada Jun
Published: July 2024

Objective:
Chronic infusion of angiotensin II (AngII) promotes development of abdominal aortic aneurysms (AAAs) in Apolipoprotein E-deficient (ApoE-/-) mice. Interleukin 33 (IL-33), known as one of IL-1 family, is rapidly released from damaged endothelial and epithelial cells. IL-33 binds to its receptor, ST2, and exerts various physiological functions through its downstream signaling. Previous studies have shown that exogenous IL-33 attenuates development of AAAs through wound healing resolution. The purpose of this study was to evaluate whether IL-33 deficiency exacerbated development of AAAs by using IL-33xApoE double deficient (IL-33-/-xApoE-/-) mice.

Methods and Results:
Male ApoE-/- and IL-33-/-xApoE-/- (8-12 weeks old) mice were assigned into saline infused group (ApoE-/- n=5, IL-33-/-xApoE-/- n=7) and AngII infused group (ApoE-/- n=19, IL-33-/-xApoE-/- n=19). Mice were infused subcutaneously with either saline or AngII by osmotic minipumps for 4 weeks. During 4-week AngII infusion, 3 mice died by AAA rupture in each AngII-infused group. There was no significant difference in ex vivo abdominal aortic width between saline infused groups (ApoE-/- 0.87±0.17 mm vs IL-33-/-xApoE-/- 0.87±0.14 mm, n.s.) nor body weight (ApoE-/- 27.3±0.9 g vs IL-33-/-xApoE-/- 27.9±0.9 g, n.s.) but serum total cholesterol concentrations in IL-33-/-xApoE-/- was higher than in ApoE-/- (522±85 mg/dL vs 427±108 mg/dL, p=0.026). AngII equally elevated systolic blood pressure (ApoE-/- 151±15 mmHg, IL-33-/-xApoE-/- 155±17 mmHg, n.s.). AngII increased ex vivo maximum abdominal aortic width in AngII infused groups, but AAAs were further exacerbated in the group of IL-33-/-xApoE-/- mice (ApoE-/- 1.31±0.09 mm vs IL-33-/-xApoE-/- 1.76±0.09 mm, p=0.001). In addition, the severity of AAAs in AngII infused groups was significantly worse in IL-33-/-xApoE-/- than in ApoE-/- (p=0.037).

Conclusion:
Our study demonstrated IL-33 deficiency exacerbated AngII-induced AAAs in male ApoE-/- mice.
Abstract 1003: Blood Pressure Metrics In Predicting Successful Thoracic Endovascular Aortic Repair
Authors: Bohr Nicole L , Alakhdar Amira , Pugar Joseph , Washburn Newell , Pocivavsek Luka
Published: July 2024

Blood pressure (BP) is a well-known risk factor for propagation of type B aortic dissections; however, there is limited data on its impact following surgical repair. While metrics such as BP variability have been associated with higher risk of cardiovascular events, such metrics has not been explored post TEVAR where management has largely focused on reducing systolic BP. By pairing unique BP metrics with preoperative aortic geometry, which has been shown to predict post-TEVAR outcomes, we can better understand what BP metrics are most useful. BP data collected in the first 24 hours after TEVAR for a cohort of 31 patients was used to calculate 24 unique BP metrics, including measures of centrality, variability, and stationarity. Preoperative CTA imaging data were used to create surface models for each aorta, which were then used to define aortic geometry as a unique point in the validated shape-size feature space. Logistic regression models with the BP and aortic geometry metrics were created to predict successful TEVAR outcomes using standard feature selection procedures and were validated using cross-validation (CV). Training and cross-validation accuracy were then compared between the models to determine the most generalizable model. A simple model using only preoperative aortic geometry in the two-dimensional shape-size feature space yielded a training accuracy of 77.4% and CV accuracy of 74.8%. An overfit model with all 24 BP metrics calculated from all available BP data (18.1 ± 6.0 BPs, range 5 - 29) had a 100% training accuracy but CV accuracy of 77.6%. Using the coefficients from this overfit model, we selected the most important features to train the next model. This resulted in a model using aortic geometry in addition to two systolic BP stationarity metrics and diastolic BP standard deviation. This model yielded a 96.8% training accuracy and 93.8% CV accuracy. Adding BP metrics creates a novel multi-dimensional aortic geometric-physiologic feature space and improves our ability to predict post-TEVAR outcomes. The BP variables that best contributed to this model were all measures of variability and stationarity, indicating that trends in BP must be evaluated to better identify patients at risk of poor outcomes.
Abstract 1009: Elevated Lipoprotein (a) Is Associated With Increased Risk Of Abdominal Aortic Aneurysm.
Authors: Sharma Pranav , Judy Renae , Yuan Shuai , Gellatly Corey , Saxby Katie , Bown Matthew J , Levin Michael G , Damrauer Scott M
Published: July 2024

Introduction:
Lipoprotein(a) (Lp(a)) is an apolipoprotein B (ApoB) containing particle that is causally associated with atherosclerotic vascular disease and the target of emerging therapeutics. Recent work has highlighted the role of circulating lipoproteins in abdominal aortic aneurysm (AAA). We sought to triangulate human observational and genetic evidence to evaluate the role of Lp(a) in AAA.

Methods:
We tested the association between levels of Lp(a) and clinically diagnosed aortic aneurysms while controlling for traditional AAA risk factors and levels of ApoB using logistic regression among 798 individuals with and 384,499 individuals without AAA in the UK biobank (UKB). Multivariable Mendelian randomization (MVMR) was used to test for putatively casual associations between Lp(a) and AAA controlling for ApoB. Genetic instruments for Lp(a) and ApoB were created from genome-wide association studies (GWAS) of Lp(a) and ApoB comprising 335,796 and 418,505 UKB participants, respectively. The instruments were tested against AAA using data from a GWAS of 39,221 individuals with and 1,086,107 without AAA.

Results:
Elevated Lp(a) levels were associated with an increased risk of AAA (OR 1.04 per 10 nmol/L Lp(a); 95%CI 1.02-1.05; P<0.01). Clinically elevated Lp(a) levels (>150nmol/L) were likewise associated with an increased risk of AAA (OR 1.50; 95%CI 1.17-1.91; P<0.01) when compared to individuals with Lp(a) levels <150nmol/L. MVMR confirmed a significant association between genetically proxied increased Lp(a) and increased genetic liability to AAA (OR 1.13 per SD increased in Lp(a); 95%CI 1.02-1.24; P<0.02).

Conclusion:
Both observational and genetic casual inference analyses support an association between increased Lp(a) and AAA risk that is independent of ApoB. Mendelian randomization analyses posit a causal relationship. These findings suggest that Lp(a) may be a therapeutic target for AAA and drive the inclusion of AAA as an outcome in clinical trials of Lp(a) antagonists.
Abstract 1011: Evolving Burden Of Aortic Aneurysm Attributable To High Systolic Blood Pressure In 204 Countries And Territories From 1990-2019: A Comprehensive Global Assessment
Authors: Goel Shrey , Vemparala Priyatha , Patel Dhruval , Vavaliya Nihar , MADHU KRUPA , Lakkimsetti Mohit , Singh Kushagrita , PATEL JUHI , AMIN VISHRANT P , DESAI HARDIK D
Published: July 2024

Introduction and Background:
High blood pressure (HBP) is a major risk factor contributing to cardiovascular disease. Understanding the global burden of aortic aneurysms (AA) attributable to HBP is crucial for healthcare professionals, researchers, policymakers, and public health experts.

Method:
Using Global Burden of Disease tool, we estimated deaths, disability adjusted life years (DALYs) due to AA attributable to HBP by age, sex, year across the 204 countries and territories from 1990-2019. The results were employed in absolute counts and age-standardized rate (per 100,000 person years).

Results:
The study observed a notable increase in the total number of deaths due to AA attributable to HBP, rising from 36,629 in 1990 (95%[UI]: 28,841-44,353) to 59,880 in 2019 (46,691-72,639). Similarly, disability-adjusted life years (DALYs) escalated from 782,721 (629,321-944,867) in 1990 to 1,241,450 (1,015,874-1,468,103) in 2019. The most significant annual percentage change (APC) in the unadjusted death rate was recorded in the high-income Asia Pacific region, with an increase of 181%, followed by a 125% increase in the DALY rate in East Asia during the period from 1990-2019. In 2019, India, Japan, Russia, China, the United States, and Brazil accounted for 49.86% of the total global deaths. The highest mortality was seen in individuals aged 75-79 years, while the highest DALYs were observed in the 65-69 year age group. Over the past three decades, males have shown higher rates of death and disability compared to females.

Conclusion:
The study shows a marked global increase in both deaths and disability from AA attributable to HBP from 1990-2019, with significant regional disparities and a higher impact on older males, emphasizing the need for targeted healthcare strategies.
Abstract 1012: Single-Cell RNA Sequencing Defines Focal Adhesion-Matrix Interactions In Smooth Muscle Cells In Thoracic Aortic Disease Triggered By BAPN Inhibiting Lysyl Oxidase
Authors: Guan Pujun , Zhou Zhen , Sarkar Ripon , Guo Dongchuan , Duan Xue Yan , Kwartler Callie , Milewicz Dianna M
Published: July 2024

Background:
Thoracic aortic aneurysms and/or dissections (TAD) are associated with pathogenic variants in genes that involved in either smooth muscle cell (SMC) contraction or extracellular matrix (ECM). The inability to maintain homeostasis between SMCs and ECM is pivotal in TAD pathogenesis. Focal adhesions (FA) act as mechanosensors in cells, transferring mechanical signals from the ECM to the intracellular contractile filaments and regulating downstream Rho/ROCK signaling. However, the landscape of SMC-ECM interactions, has not been fully explored at the early stages of TAD.

Methods:
C57BL/6J wild-type mice aged 21 days postnatal (P21) were administered the ECM cross-link blocker, BAPN, for up to four weeks to induce TAD. Proximal aortas were harvested for single-cell RNA sequencing at P35. Immunofluorescence and immunoblots were used to validate findings.

Results:
Single-cell RNA sequencing classified SMCs into 5 clusters marked by
Rgs5
(SMC

Rgs5

+
), high levels of
Palld
(SMC

Palld

+
),
Tnnt2
(SMC

Tnnt2

+
, marker of second heart field-derived cells),
Atf3
(SMC

Atf3+

cluster), or no clear marker (SMC
NOS
). The analysis of how BAPN exposure affects different cell types, based on the Jensen-Shannon divergence in the probability distribution between groups, demonstrated that SMCs, and specifically SMC

Atf3

+
and SMC
NOS
, are the most perturbed cell cluster by BAPN. Gene set enrichment analysis identified significantly dysregulated pathways of FA and Rho/ROCK signaling. The expression of
Itga5
and
Fn1
(fibronectin) were increased and
Itga8
was decreased. An SMC-SMC interaction analysis found
Fn1
interactions were the most upregulated interaction in 4 SMC clusters. When analyzing downstream pathways due to the changes in interaction, 80% of the top 25 interactions were related to integrins, including increased Fn1 interactions with Itga5-Itgb1 and Itgav-Itgb3. In SMC

Rgs5

+
cluster, the most affected interactions are related to TGF-β signaling.

Conclusions:
These findings suggest that, in the majority of aortic SMCs, mechanosensing through FAs, driven by fibronectin interactions with specific integrin receptor pairs, is activated with BAPN treatment, potentially contributing to the pathogenesis of TAD.
Abstract 1026: Apolipoprotein A1 On High-density Lipoprotein Interferes With Low-density Lipoprotein Binding To Proteoglycans
Authors: Geh Esmond , Roscoe Isabella , Swertfeger Debi K , Bursey Alexiana , Tarapore Pheruza , Sexmith Hannah , Davidson William , Shah Amy S
Published: July 2024

Background:
The entrapment of apolipoprotein (APO) B-100 containing low density lipoproteins (LDL) by proteoglycans (PGs) in the extracellular matrix (ECM) of the arterial intima is a key initial step in the development of atherosclerotic vascular lesions. High density lipoproteins (HDL) can interfere with this process, but the underlying mechanism is not fully understood and is thought to be due to APOE. The aim of this study was to utilize our recently developed quantitative assay that measures the binding of LDL to PGs in the ECM and to investigate how HDL and other apolipoproteins may influence this process.

Methods:
An in-cell ELISA was used to measure the binding of LDL to PGs in the ECM synthesized by mouse vascular smooth muscle cells. Fast Protein Liquid Chromatography, immunoprecipitation, and immunoblotting analysis were performed to identify how HDL interacts with LDL to prevent binding to ECM-PGs.

Results:
LDL binding to PGs was inhibited by HDL and APOA1 in a dose dependent manner. Competition experiments showed that HDL does not compete directly with LDL for ECM-PG binding. Instead, our data revealed an association between LDL with APOA1 that diminishes LDL ability to bind ECM-PGs.

Conclusion:
HDL inhibits LDL binding to ECM-PG through an interaction with its main apolipoprotein, APOA1 that is different from APOE. Further investigation is warranted to understand the role of this interaction in the atherosclerosis process.
Abstract 1033: Loss Of Serine Hydroxymethyltransferase 2 In Macrophages Blunts Glutathione And Enhances Ceramide Synthesis To Accelerate Atherosclerosis
Authors: Rohilla Sumati , Ghrayeb Alia , C Finney Alexandra , McKinney M. Peyton , Liu Yuhao , Nam Hugh , Tan Lin , L Lorenzi Philip , Mor Inbal , Orr A Wayne , Gottlieb Eyal , Yurdagul Arif , Rom Oren
Published: July 2024

Increasing evidence suggests an important role for amino acid metabolism in atherosclerotic cardiovascular disease (ASCVD), highlighting potential biomarkers, causative factors, and therapeutic targets. Among all amino acids, lower circulating glycine is consistently reported to be associated with enhanced ASCVD severity. The amino acid serine is the main precursor for glycine, and a lower glycine:serine ratio is associated with more advanced and unstable atherosclerotic plaques. The mitochondrial serine hydroxymethyltransferase 2 (SHMT2) catalyzes the bulk of glycine formation from serine, and is critical in development since global knockout mice are nonviable. We found that SHMT2 is highly expressed in macrophages in both human and mouse atherosclerotic plaques. LPS-treated macrophages displayed reduced SHMT2 expression concomitant with a lower intracellular glycine:serine ratio. To study the role of SHMT2 in macrophages, we developed novel myeloid-specific
Shmt2
knockout mice (
Shmt2
MKO
). Metabolomics and stable isotope tracing studies revealed that macrophages lacking SHMT2 have a lower glycine:serine ratio leading to attenuated
de novo
synthesis of glutathione and enhanced formation of the ceramide precursor, sphinganine. Accordingly, the loss of SHMT2 in macrophages enhanced superoxide production and ceramide accumulation. To determine the role of macrophage SHMT2 in atherosclerosis, we crossed the
Shmt2
MKO
mice with apolipoprotein E-deficient (
Apoe
-/-
) mice. After 12 weeks on Western diet,
Shmt2
MKO
/
Apoe
-/-
mice showed no differences in plasma lipid or amino acid profiles as compared to
Shmt2
flox
/
Apoe
-/-
mice. Nevertheless, plaque area and necrotic core size were significantly enhanced in
Shmt2
MKO
/
Apoe
-/-
mice. Taken together, macrophage SHMT2 plays a critical role in atherosclerosis by regulating redox and ceramide metabolism.
Abstract 1035: C/EBPα Has Depot-specific Roles In White Adipose Tissue Development And Function.
Authors: Hu Krista , Matias Caio , Gomez Juliana , Dodge Esme , Bauer Robert C
Published: July 2024

Obesity, defined as excess white adipose tissue (WAT), associates with a higher risk of metabolic disease and comorbidities, and continues to be on the rise in the developed world. However, different WAT depots associate with differing levels of cardiometabolic risk. Expansion of visceral WAT (vWAT) correlates with increased risk for cardiovascular disease and type 2 diabetes, while expansion of subcutaneous WAT (sWAT) does not. These observations suggest mechanistic differences between the two depots, and elucidating these mechanisms may provide insights into promoting cardiometabolic health in humans. Human GWAS have shown that SNPs near the gene
CEBPA
associate with waist-to-hip ratio, a metric of vWAT mass in humans, suggesting
CEBPA
plays a specific role in vWAT.
CEBPA
encodes the transcription factor C/EBPα, a known regulator of adipogenesis. We established adipocyte-specific
Cebpa
knockout (Cebpa_ASKO) mice and found they lack gonadal WAT (gWAT), the largest vWAT depot in mice. Conversely, sWAT is still present in Cebpa_ASKO mice, although with reduced mass (-23%, p<0.05) and larger adipocytes. Additionally, Cebpa_ASKO mice have reduced lipolysis (-26% NEFAs, p<0.05), plasma adiponectin (-89%, p<0.0001) and leptin (-82%, p<0.05), suggesting the remaining sWAT is dysfunctional. Together, these data suggest that C/EBPα has WAT depot-specific roles, being required for vWAT development, but only for function in mature sWAT. Given the myriad roles for vWAT in metabolism and the absence of vWAT in Cebpa_ASKO mice, we investigated downstream metabolic consequences of this phenotype. Cebpa_ASKO mice have increased brown adipose tissue (BAT) mass (+118%, p<0.001) due to increased ectopic lipid deposition. Cebpa_ASKO mice also have increases in plasma cholesterol (+20%, p<0.05), liver mass (+24%, p<0.01), and hepatic fat content (+329%, p<0.01). Overall, these results demonstrate that adipocyte C/EBPα significantly influences whole body metabolism. Ongoing studies center on delineating the different mechanisms through which C/EBPα functions in vWAT and sWAT. Understanding the depot-specific mechanisms of C/EBPα could have practical implications for new clinical targets of obesity and metabolic disease.
Abstract 1041: Loss Of The ETS Transcription Factor ERG Disrupts Fate-defining Programs In The Aortic Endothelium And Promotes Expansion Of Endothelial Lineage Cells In Atherosclerotic Plaque
Authors: Botts Steven R , Schulz Kristen , Scipione Corey A , Raju Sneha , Breda Leandro C , Prajapati Kamalben , Yu Kai , Khan Aniqa , Polenz Chanele K , Hyduk Sharon J , Wythe Joshua D , Robbins Clint S , Cybulsky Myron I , Fish Jason E , Howe Kathryn L
Published: July 2024

Aim:
The ETS transcription factor ERG has been identified as a principal regulator of endothelial function through its ability to repress inflammation in endothelial cells (ECs), and loss of ERG induces endothelial to mesenchymal transition (EndMT) in fibrotic disease. To investigate ERG function in atherosclerosis, we employed an EC-specific
Erg
knockout (
Erg
EC
-
KO
) in PCSK9-overexpression and apoE-deficient murine atherosclerosis models.

Methods:
Atherosclerosis was induced in 10-week-old
Erg
EC
-
KO
mice or wild-type counterparts that had a Cre-inducible EC lineage tag (
Cdh5
CreER
;
R26-CAG-LSL-Sun1-sfGFP
). Atherosclerosis was modeled by
Apoe
deletion or AAV8-PCSK9 with 12 weeks of high-cholesterol diet (1.25%). Aortic intimal cells were assessed with flow cytometry. Plaque burden, morphology, and cellularity were characterized with brightfield, Oil Red O, H&E, Movat, and immunofluorescence imaging.

Results:
Erg
EC-KO in PCSK9-overexpression mice led to a 1.6-fold increase in aortic plaque burden and plaque formation in regions that are usually protected (e.g., greater curvature; p≤0.03; n=9-10). Loss of ERG increased elastin breaks (1.7-fold; p=0.02; n=9) and decreased fibrous cap thickness (0.76-fold; p=0.02; n=9-10) in aortic arch cross sections. Aortic intimal digests from
Erg
EC-
KO
mice contained increased Sun1-sfGFP
+
EC lineage cells (1.9-fold; p=0.01; n=4), which had reduced CD31 positivity (0.79-fold; p=0.03; n=4). Notably, a 5.3-fold increase in EC lineage cells was observed in plaques from brachiocephalic artery cross sections (p=0.0001; n=6-15), which had hallmarks of EndMT (e.g., 31.5-fold increase in GFP
+
ACTA2
+
cells, p=0.02, n=3-5). Increased aortic plaque burden was recapitulated in apoE-deficient
E
rg
EC-
KO
mice (2.1-fold; p=0.02; n=4-8).

Conclusion:
Endothelial
Erg
KO results in increased plaque burden and altered plaque topography in murine atherosclerosis. Loss of ERG induces EndMT and marked ingression of ECs from the luminal endothelium to the plaque interior. These findings highlight loss of ERG as a novel mechanism by which intimal ECs acquire mesenchymal identity in atherosclerosis and implicate EC identity loss as a driver of EC migration into plaques, which is associated with a vulnerable morphology.
Abstract 1046: Miro1 Promotes Mitochondria ER Contact Sites Formation In Vascular Smooth Muscle Cells During G1/s
Authors: Endoni Benney , Koval Olha , Qian Lan , Grumbach Isabella M
Published: July 2024

Vascular smooth muscle cell (VSMC) proliferation plays a key role in vascular disorders including atherosclerosis. Mitochondria promote VSMC proliferation by mitochondrial Ca
2+
regulation, ROS and ATP production. Mitochondria-ER contact sites (MERCS) promote signal transduction to mitochondria. MIRO1, an outer mitochondrial membrane GTPase, drives intracellular mitochondrial mobility. In drosophila, MIRO1 controls MERCS formation. However, it is unknown whether this interaction promotes key cellular eukaryotic phenotypes, such as proliferation. Here, we tested whether MIRO1 promotes VSMC proliferation by modulating MERCS and mitochondrial Ca
2+
entry. MIRO1 deletion decreased neointima formation in vivo, and abolished VSMC proliferation and G1/S transition in vitro. In WT VSMCs, proximity ligation assays (PLA) and split-GFP imaging revealed that MERCS formation increased in G1/S phase along with recruitment of MIRO1. MIRO1 deletion abolished cell cycle-dependent regulation of MERCS while increasing the expression of MERCS proteins GRP75 and IP3R. MIRO1 deletion also blocked increases in baseline mitochondrial [Ca
2+
] by mitoPericam imaging and decreases in ER [Ca
2+
] by CEPIA-ER in G1/S phase. Additionally, the mitochondrial membrane potential (TMRM) and mitochondrial respiration significantly increased at G1/S in WT VSMCs but not in MIRO1-/- cells. In WT VSMCs, pyruvate dehydrogenase activity increased G1/S phase while cytosolic ATP levels (ATP sensor) were increased. In MIRO1-/- cells, both parameters did not change. These data demonstrate that MERCS formation and mitochondrial [Ca
2+
] increases during the cell cycle and is controlled by MIRO1. Furthermore, MIRO1 deletion decreases cellular ATP levels in VSMCs. In conclusion, we propose that MIRO1 controls MERCS and increased mitochondrial [Ca
2+
] that drives metabolic activity and proliferation.
Abstract 1058: Effects Of Soluble Guanylate Cyclase Activation On Progression Of Fibrocalcific Aortic Valve Stenosis (avs), Its Molecular Drivers, And Downstream Pathological Cardiac Electrical Remodeling
Authors: Dubey Saumya , Roos Carolyn M , Huang Runqing , Verzosa Grace C , Smith Leslie , Hagler Michael A , Zhang Bin , Miller Jordan D
Published: July 2024

Objective:
We recently discovered that activation of oxidized soluble guanylate cyclase with ataciguat (ATA) slows valve calcification and dysfunction in mice and humans with moderate aortic valve stenosis (AVS). We examined if ATA would attenuate multiple mechanisms contributing to AVS and end organ dysfunction associated with AVS, and hypothesized: 1) ATA attenuates osteogenic signaling in valve tissue from mice with AVS, thereby attenuating calcification and valve dysfunction, 2) AVS increases rates of sudden cardiac death in our model, and 3) treatment with ATA prevents QT prolongation and deleterious electrical remodeling commonly observed in AVS.

Methods:
Functional, molecular, and histopathological changes in valve tissue along with QT interval duration were measured in 3 independent groups of mice: non-AVS, AVS, and AVS+ATA (WD for 6 months then WD+ATA for 3 months (AVS+ATA). We gave 5 mg/kg isoproterenol (ISO, i.p.) to ldlr
-/-
/ApoB100-only mice fed normal chow (non-AVS) or western diet (AVS, fed WD for 9 months) to assess risk for malignant arrhythmias. Retrospective analysis of QT interval was also done on data from our recent Phase 1 clinical trial.

Results:
AVS+ATA mice had less valve calcification and slower progression of valve dysfunction through attenuation of canonical BMP signaling and reductions in Runx2 and SPP1 compared to AVS mice. QT prolongation was evident in AVS mice, and was significantly attenuated by treatment with ATA (p= 0.001) and coarsely associated with severity of valvular or ventricular function. While all non-AVS mice survived for 24 hours after ISO, 40% of mice with AVS reached a moribund equivalent within 4 hours. Retrospective analyses also showed reductions in QT interval duration in patients treated with 200 mg/day ATA for 14 days (p < 0.05).

Conclusion:
Collectively, this is the first study reporting increased propensity for malignant arrhythmias in our mouse model of AVS, QT prolongation as a likely underlying mechanism, and restoration of cardiac conduction dynamics with targeted restoration of nitric oxide signaling. Future studies focused on the role of sGC in the evolution and prevention of AVS and consequent deleterious electrical remodeling/malignant arrhythmia risk is warranted.
Abstract 107: Effects Of Immune-checkpoint Inhibitors On Atherosclerosis In Halted Progression Model
Authors: Cilhoroz Burak T , Das Dayasagar , Amadori Letizia , Eberhardt Natalia , Smyrnis Panagiotis , Gourvest Morgane , Gildea Michael , Kaur Ravneet , Moore Kathryn J , Giannarelli Chiara
Published: July 2024

Immune checkpoint inhibitor (ICI) treatment in cancer has been linked to atherosclerotic-related cardiovascular events. While ICIs have been shown to aggravate atherosclerosis in mouse models of plaque progression, these models do not reflect the clinical scenario of cancer patients with pre-existing atherosclerosis under lipid-lowering treatments. To address this, we tested the effects of PD1 or CTLA4 inhibitors (aPD1 or aCTLA4), and their combination (aPD1 + aCTLA4), in a model of halted atherosclerosis progression, mimicking standard of care lipid-lowering treatments.
Ldlr
-/-
mice (n=126) mice were fed a western diet (WD) for 17 wk (baseline), followed by a switch to standard chow diet for 4 wk (halted progression), and mice were randomized to treatment arms. Histological and single cell RNA sequencing (scRNA-seq) revealed that inhibiting PD-1 or CTLA-4 individually aggravated atherosclerosis, despite comparable cholesterol reduction to isotype control groups. Surprisingly, the combination of aPD1 + aCTLA4 did not show the same proatherogenic effect observed individually. Distinct effects on plaque composition were noted, with aPD-1 or aCTLA4 leading to increased plaque collagen content, and their combination showing plaque stabilization with higher cap thickness and similar collagen content to controls. Immunostaining revealed no differences in plaque macrophage content among groups, but aPD-1-treated mice exhibited increased CD8 T cells. scRNAseq showed that aPD1 increased specific inflammatory myeloid clusters, including Spp1+ macrophages and Ccr2+ MoDCs, and reduced anti-inflammatory IL16+ DPTs but increased cytotoxic Gzmk+ CD8+ EMRA T cells. CTLA4 inhibition increased foamy Trem2hi macrophages and Ccr2+ MoDCs. Yet, combined aPD1 + aCTLA4 did not significantly alter cell proportions compared to halted progression. In conclusion, our study demonstrates that inhibiting either PD1 or CTLA4 individually aggravates atherosclerosis, under cholesterol lowering conditions. These findings highlight the nuanced effects of immune checkpoint inhibitors on atherosclerosis, emphasizing the potential importance of considering combination therapies and lipid lowering for better outcomes in cancer survivors.
Abstract 1108: Sex And Racial Differences In The Outcomes Of Intravascular Ultrasound-guided Percutaneous Coronary Intervention: A Five-year Retrospective Analysis
Authors: Evbayekha Endurance O , Nriagu Bede N , Idowu Abiodun , Dixon Brittany , LaRue Shane J , Reiss Craig
Published: July 2024

Introduction:
Studies have shown the superiority of intravascular ultrasound-guided percutaneous coronary intervention (IVUS-PCI) over angiographic PCI alone. We studied the racial and sex differences in outcomes.

Methods:
Using the National Inpatient Sample database (2016-2020) and ICD-10 codes, we identified the IVUS-PCI population. Chi-square and student T-tests were used to compare categorical and mean differences. Multivariate logistic regression analysis was used to account for potential confounding variables such as sex, age, comorbidities, hospital size, and teaching status and to estimate the probability of outcomes. Our primary outcome was mortality. A p≤0.05 was significant.

Results:
Of the 271,530 PCI hospitalizations, 12% (32,675) got IVUS-PCI (63.7 years ± 11.7), of which 30.4% (9,925) were females. The probability of receiving IVUS-PCI was similar among all races: 1.04 (0.9-1.1, p=0.7) and sex 1.01 (0.9-1.03, p=0.6). Compared to Whites, Blacks 0.8 (0.6-1.2), Hispanics 0.9 (0.7-1.4), and Asians 1.3 (0.8-2.2), (p>0.05, each) did not differ in mortality. Also, no sex differences in mortality, with females 1.2 (1.0-1.5, p>0.05). Post-procedural shock 2.5 (1.5-4.0), bleeding 1.8 (1.3-2.6), and anemia 1.5 (1.3-1.8) (p<0.05, each) were more likely in females. No racial differences were noted.

Conclusion:
In our IVUS-PCI population, there was no difference in mortality among race or sex. However, females were more likely to suffer peri and post-procedural complications. More research is needed to identify the drivers of these disparities.
Abstract 1117: Il-1 Signaling On Vascular Smooth Muscle Cells Accelerates Atherosclerosis After Murine Kawasaki Disease Vasculitis
Authors: Chen Shuang , Savas Asli , Aubuchon Emily , Lee Youngho , Lane Malcolm , Moreira Debbie , Shimada Kenichi , Crother Timothy R , Noval Rivas Magali , Arditi Moshe
Published: July 2024

Objective:
We have previously shown that
Lactobacillus casei
cell wall extract (LCWE)-induced Kawasaki disease (KD) vasculitis significantly accelerates atherosclerosis in hypercholesterolemic mice on high fat diet. IL-1 signaling is known to play a key role in both KD vasculitis and in the development of atherosclerosis. Here, we investigated the contribution of IL-1 signaling on vascular smooth muscle cells (VSMCs) in KD vasculitis-induced acceleration of atherosclerosis.

Methods:
Tamoxifen-inducible VSMC-specific IL-1 receptor (
Il1r1)
knockout (

Myh11
Cre

-ERT2

Il1r1
Δ/Δ

) mice and

Il1r1
fl/fl

littermate controls, all on ApoE
-/-
background, were injected with either PBS or LCWE. Following induction of KD vasculitis for 2 weeks, mice were fed a tamoxifen diet for 2 weeks to induce
Il1r1
deletion on VSMCs, before being exposed to 8 weeks of Western diet to promote atherosclerosis.

Results:
KD vasculitis was associated with a significant acceleration of atherosclerosis, as expected.

Myh11
Cre

-ERT2

Il1r1
Δ/Δ

mice had significantly diminished atherosclerotic plaque size, lipid composition, macrophage infiltration and necrotic core formation in aortic root as well as diminished lipid accumulation in aorta en face measurements compared with

Il1r1
fl/fl

control mice despite similar cholesterol levels. We also observed that

Myh11
Cre

-ERT2

Il1r1
Δ/Δ

mice had significantly diminished endothelial adhesion molecules VCAM-1 and ICAM-1 expression in the lesion area and serum monocyte chemotaxis protein-1 (MCP-1) level compared with

Il1r1
fl/fl

control mice, consistent with the reduction of macrophage infiltration that we observed.

Conclusions:
Our results suggest an important pathophysiologic link between IL-1 signaling, specifically on VSMCs, and subsequent acceleration of atherosclerosis in hypercholesterolemic mice following KD vasculitis. Thus, further studies are warranted on the role of IL-1 signaling not only in acute KD, but also in the subsequent vascular remodeling and complications following acute KD vasculitis including atherosclerosis.
Abstract 1120: Untargeted 16s Ribosomal Rna Survey Of Microbial Communities In The Patient With Aneurysmal And Occlusive Aortic Disease
Authors: Geiger Joshua T , Gill Ann , Kedwai Baqir , Lehane Daniel , Ford Benjamin , Kruger Joel , Stoner Michael , Gill Steven , Mix Doran S
Published: July 2024

Introduction:
Inflammation is a hallmark of aneurysmal disease, but the etiology for the inflammation is unknown. Previously, bacteria and bacterial nucleic acid sequences have been identified within abdominal aortic aneurysm tissue. However, only one study has taken an un-targeted approach to assess the diversity of species and no study has surveyed bacterial species across the various environments in patients with aortic disease.

Research Question:
Does the distribution of bacterial sequences differ between environments and by disease status in patients with abdominal aortic disease?

Methods:
Whole blood, acute (luminal) thrombus, chronic (medial/abluminal) thrombus, and aortic wall specimens were obtained from 14 patients undergoing open aortic surgery. Five patients had aneurysm disease and nine patients had aortic occlusive disease. Sequencing using universal primers targeting the V3 - V5 16s rRNA bacterial gene was performed. Reads were processed and aligned to reference banks using QIIME2 to the genus level.

Results:
The average patient age was 60.9 years with no significant difference in baseline demographics. Phyllobacterium was the most common genus identified in acute and chronic thrombus for aneurysmal and occlusive disease while pseudomonas was the most common genus identified in aortic wall. Chao1 alpha diversity was higher in aneurysmal aortic wall samples compared to occlusive disease (10.6 vs 6.6, p=0.047). On principle component analysis, samples from similar tissue environments cluster together with 16% of the sequence variation explained by the grouping (p=0.003, Figure 1).

Conclusions:
The diversity of bacterial sequences groups by microenvironment within patients with aortic disease, suggesting a spectrum of bacterial communities that progress from blood through the luminal thrombus to the aortic wall. Further work could distinguish genus level variations that associates with disease in the different environments.
Abstract 1124: Sympathetic Denervation Models In Murine Femoral Arteries: Acute To Chronic Effects Of 6-hydroxydopamine
Authors: Brown Taylor K , Dang Caitlyn , Xiong Liqun , Alharbi Sara , Smires Aidan , Ho Karen J , Jiang Bin
Published: July 2024

Introduction:
The autonomic nervous system (ANS) drives blood flow regulation in peripheral arteries. This study aims to develop novel models of sympathetic denervation in peripheral arteries and to characterize subsequent vascular remodeling. We hypothesize that sympathetic denervation of murine femoral arteries can be achieved through local administration of neurotoxin 6-hydroxydopamine (6-OHDA).

Methods:
BALB/c mice underwent local administration of 6-OHDA to the left femoral artery, while the contralateral artery received vehicle solution. Two approaches were tested: (1) Acute: a one-time topical application following surgical exposure of femoral arteries (N=8, male,
Fig. A
), and (2) Chronic: weekly subcutaneous injections near the femoral arteries (N=11, male and female,
Fig. B
). Hindlimb perfusion was assessed with laser Doppler imaging, and arterial structure and remodeling were evaluated with histology and immunohistochemistry at one, two, or four weeks.

Results:
One-time 6-OHDA application (Acute) led to temporary femoral artery denervation (reduction in tyrosine hydroxylase) for one week (
Fig. C – D
), with inflammation observed in both 6-OHDA and vehicle-treated arteries. No vascular wall remodeling was observed after temporary denervation. Weekly subcutaneous 6-OHDA injections (Chronic) resulted in persistent denervation throughout the four-week period in male mice, while female mice displayed variable outcomes (
Fig. E – F
). Sustained sympathetic denervation in the chronic model led to reduced hindlimb blood perfusion at low core temperatures (
Fig. G – H
), though no significant differences were detected in vascular smooth muscle cell phenotype or extracellular matrix composition.

Conclusions:
Local administration of 6-OHDA is an effective means of peripheral artery sympathetic denervation with controllable duration. This innovative approach of targeting the ANS unveils new prospects for advancing vascular therapies.