bluuebunny/crossref_metadata_2025 huggingface dataset:
The Morality of The Prosecutor (Pancasila Moral Relations Guarantee Law Enforcement in Political Determination)
Authors: Ash-shidiqqi M. Ellectrananda Anugerah , Zaelani Aziz
Published: July 2024

The purpose of this study is to determine the form of Pancasila's moral relationship as Indonesia's legal ideology in defending the prosecutor's office in the midst of political determination. In this work, a conceptual method is employed as a foundation for thinking to help answer problems that will subsequently be qualitatively investigated. The study's findings suggest that Pancasila, as Indonesia's legal ideology, can protect the prosecutor's office through moral relationships, which are reflected in the prosecutor's integrity. The consequence is that in order to achieve the objectives of justice in law enforcement, it is essential to maximize the implementation of the prosecutor's office's integration with a Pancasila worldview. Pancasila values can influence the prosecutor's office's integrity by allowing them to use their roles and powers to counter political determinism in the form of government interference.
Unravelling the User Perception: Evaluating Kenadsa's Heritage Office Building Performance Through Perceptual Dimensions and Behaviors
Authors: Hamlili Fatima Zohra , Dakhia Azzedine , Biara Ratiba Widad
Published: July 2024

This study investigates the complex relationship between user perceptions and subsequent behaviors in the multisensory environment within Kenadsa’s heritage office building, located in the oasis of Bechar, Algeria. Using a combination of quantitative and qualitative subjective data collection methods, the research aims to assess the perceptual dimensions of the office building's interior space, thermal conditions and lighting environment, as well as to evaluate its performance based on users' perceptions and behaviors. The methodology includes in situ questionnaire surveys conducted after each user's work experience, complemented by thermal and visual assessments observation of the lighting during the winter and summer seasons. The results highlight the dynamic interplay between user perception and behavior, and provide valuable information for improving the design and functionality of heritage buildings in similar contexts.
Representation of Victim and Perpetrator in Adelina Lisao’s Abuse Case in Malaysian Online News Articles
Authors: Binti Adanan Aizatul Alyaa , Binti Ungku Mohd Nordin Ungku Khairunnisa
Published: July 2024

This research analyses the representation of victim and perpetrator as social actors and its argumentation strategies from Malaysian online news articles surrounding the tragic case of Adelina Lisao, an Indonesian domestic worker who lost her life in Malaysia due to abuse by her employer. Employing a qualitative research approach, the data is systematically collected with keywords to identify relevant news articles pertinent to the case: "Adelina Lisao” and “Ambika Shan”. This research utilises the Social Actor’s Network (SAN) and the Critical Discourse Analysis (CDA) framework, specifically the Discourse-Historical Approach (DHA), to examine 39 online news articles about the case from the most popular Malaysian news website in 2023, Malaysiakini. The findings revealed that Adelina Lisao was mostly represented as a passive receiver of the abuse, while Ambika Shan’s role as a perpetrator was mostly backgrounded. The findings contribute to a better understanding of how social actors are portrayed on online websites and the argumentation strategies within the context of domestic worker abuse cases in Malaysia. Thus, it acknowledges the importance of language in online news discourse in shaping public perceptions, particularly regarding domestic workers in Malaysia as evidenced by the representation of victim-perpetrator dynamics in such cases.
A história dos irmãos gêmeos Tomaz e Gabriel: contada por Wanise Martinez
Authors: Martinez Wanise ,   Universidade de São Paulo. Instituto de Psicologia, Otta Emma ,   Universidade de São Paulo. Instituto de Psicologia
Published: July 2024

Histórias pessoais de gêmeos nem sempre chegam a quem poderia se beneficiar do estudo desses eventos, como profissionais da psicologia e leitores interessados. Descobertas quantitativas de estudos de gêmeos criados separados nos dizem muito sobre os fatores genéticos e ambientais que afetam nossas características comportamentais, físicas e médicas, mas mascaram a riqueza das pessoas por trás delas. É importante entender o significado da ligação entre gêmeos idênticos. Este trabalho é um relato da trajetória de gêmeos idênticos separados aos quinze dias de vida e reunidos aos 23 anos. O livro está organizado em dez capítulos que levam os leitores à jornada única de Tomaz e Gabriel, partindo do reencontro e voltando ao momento quando suas vidas foram separadas, mostrando suas diferentes famílias e comunidades, sua espiritualidade e sexualidade, a fusão e a divergência de suas vidas atuais.
O canto fala mais: da acústica à cognição
Authors: Medeiros Beatriz Raposo de ,   Universidade de São Paulo. Faculdade de Filosofia, Letras e Ciências Humanas
Published: July 2024

Por que ler este livro? O objetivo dessa obra é mostrar um percurso de estudos sobre fala e canto a partir da fonética, que é uma subárea da linguística, utilizando uma linguagem que, tanto leigos como acadêmicos de diversas áreas, compreendem.  Nesse sentido, conhecimentos sobre os sons do canto comparativamente à fala são apresentados, tratando da acústica e da articulação em relação à produção da fala e do canto, que é considerado um tipo de fala. O canto é tratado como um assunto que pode ser estudado pela ciência da linguagem, a linguística. Também pode ser estudado a partir de paralelos traçados entre ritmo de fala e ritmo da música. Atar tais pontas, neste livro, significa tornar acessível muitos aspectos e elementos que são estudados na universidade e que devem ser amplamente divulgados para toda e qualquer pessoa interessada.
O ensino e a aprendizagem não cabem em algoritmos: relato docente sobre o fetiche da inteligência artificial
Authors: Figueira Monique
Published: July 2024

This is a teaching experience report on the “Ethics and information” course at a federal university. Its first section describes its theoretical foundation, based on Álvaro Vieira Pinto’s cybernetics and the political economy of information and communication. Its second section discusses the course participatory methodology inspired by Paulo Freire (Vieira Pinto’s student), which resorted to multimedia didactic, self-assessment, dozens of student seminars, and a final open essay. As its main result, dialogic education confirms students’ unreasonable fascination with texts generated by artificial intelligence, along with the unpreparedness of the institution to address this new challenge.
A sala de aula extramuros: o impacto da curricularização da extensão na graduação de Jornalismo
Authors: Saback Lilian
Published: July 2024

This study investigates the impacts of including extension credits in undergraduate courses on the training of journalists. It describes the work carried out by PUC-Rio Journalism and Citizenship students at the Fala Roça newspaper, in Rocinha, a favela in southern Rio de Janeiro. The theoretical research is supported by the concepts of the “community that comes” (Agamben, 1993) and the “external agent”, by the Canadian philosopher Kenneth Schmitz, as Raquel Paiva described in her work Espírito Comum: comunidade, mídia e globalismo (2003), which addresses relationships established by the partnership and the development of “responsibility” proposed by Mikhail Bakhtin (1895-1975) in the act of producing citizen journalism. This research shows that university extension enables the journalism discipline to train journalists who have “responsibility” and can effectively be social agents by transforming the society in which they live.
Desenvolvendo capacidades para a comunicação democrática: uma abordagem integrada de ensino baseada na deliberação
Authors: Maia Rousiley Celi Moreira , Hauber Gabriella , Leão Augusto Veloso , Cunha Leonardo Santa Inês
Published: July 2024

This study describes an innovative educational proposal to develop deliberative skills – motivation and ability to explain points of view based on reflected considerations, reciprocal involvement, and more inclusive and respectful communication. Based on theories of deliberative democracy – and expanding citizen participatory initiatives and experiences in the area of education, we propose an approach with (i) peer learning, (ii) playful and performative activities, and (iii) authentic discussions about controversial political issues. Our experiment illustrates how to develop a conceptual understanding of deliberative principles, promote selfreflection, and practise democratic discussions on contentious issues. This study has theoretical and practical implications for programs that seek to promote democratic communication.
Abstract 103: Acly Inhibition As A Novel Therapeutic Approach For Vascular Remodeling In Coronary Artery Disease
Authors: Grobs Yann , Romanet Charlotte , Lemay Sarah-Eve S , Bourgeois Alice , Theberge Charlie , Sauvaget Melanie , Breuils-Bonnet Sandra , Martineau Sandra , El Kabbout Reem , Potus Francois , Voisine Pierre , Provencher Steeve , Boucherat Olivier , Bonnet Sebastien
Published: July 2024

Coronary artery (CoA) disease (CAD) is characterized by neointimal hyperplasia (NH), which is driven by a cancer-like pro-proliferative phenotype of smooth muscle cells (SMCs). A metabolic shift towards glycolysis and changes in the epigenetic landscape contribute to this abnormal phenotype. ATP Citrate Lyase (ACLY) is an enzyme that plays a role in promoting the Warburg effect, lipid synthesis, and chromatin remodeling in cancer. However, its role in CAD remains unknown. We hypothesized that ACLY is upregulated in CAD and supports the abnormal phenotype of CAD-CoASMCs. To investigate this hypothesis, we measured ACLY expression in human distal coronary arteries and isolated CoASMCs from CAD patients. We found increased expression and activation of ACLY in CAD-CoASMCs, with preferential localization in the nucleus. Inhibition of ACLY using BMS-303141 or siACLY decreased CAD-CoASMCs proliferation and survival. ACLY inhibition also reduced glycolysis markers and increased mitochondrial respiration. Additionally, ACLY inhibition lowered cholesterol levels and lipid droplet accumulation in CAD-CoASMCs. Further analysis showed that ACLY promotes nuclear acetyl-CoA production, leading to acetylation of specific histone proteins and GCN5-mediated transcriptional activation of genes involved in cell cycle progression. The transcription factor FOXM1 appeared to mediate this genetic signature.
In vivo
, pharmacological inhibition of ACLY using BMS-303141 attenuated NH in a rat model of carotid injury (CI). SMC-specific
Acly
K.O mice were protected against CI-induced NH. Consistently, tamoxifen-induced SMC-specific
Acly
deletion mitigated NH in mice subjected to CI. The therapeutic effects were associated with reduced histone acetylation. To assess the translational potential of our findings, we tested the effects of ACLY inhibition on ex vivo human coronary artery and saphenous vein explants stimulated with growth factors cocktail. We found that ACLY inhibition prevent NH development, supporting the potential of ACLY as a therapeutic target. In conclusion, our study reveals that ACLY plays a critical role in vascular remodeling in CAD and its pharmacological inhibition may represent a novel avenue as therapeutic treatment.
Abstract 1048: High Density Lipoprotein To Macrophage Free Cholesterol Influx And Transfer To Low Density Lipoprotein - New Assays For Dysfunctional High Density Lipoprotein: The Houston Heart Study
Authors: Gillard Baiba K , Yelamanchili Dedipya , Saleem Hira , Gotto Antonio M , Remaley Alan , Rosales Corina , Pownall Henry J
Published: July 2024

Free cholesterol (FC) flux between HDL and macrophages switches from efflux to influx as HDL-particle number (HDL-P) and mol% FC increase. Among normolipidemic subjects, ASCVD is more prevalent among patients with HDL that are dysfunctional acceptors of macrophage-FC efflux and among patients with a very high plasma HDL concentration, i.e., hyperalphalipoproteinemia. HDL receptor deficient mice (Scarb1
-/-
), a robust model of hyperalphalipoproteinemia, are ASCVD-susceptible. Scarb1
-/-
and wild-type mouse HDL are equally good acceptors of macrophage-FC efflux but Scarb1
-/-
HDL are more
potent donors of FC influx
. This provoked our hypothesis that high HDL-FC bioavailability = HDL-P x HDL-mol% FC is associated with excess human ASCVD. Here we describe assay-validation for HDL-FC influx into macrophages and transfer to LDL.

Methods:

HDL-[
3
H]FC Transfer to J774 Macrophages:

HDL-[
3
H]FC from five volunteers were co-incubated with cells for 2 h at 37°C, cells were washed, lipid-extracted, and the extract β-counted. Influx was expressed as pmol FC uptake/mg cell-protein.

HDL-[
3
H]FC Transfer to LDL:

LDL and HDL-[
3
H]FC, each at 0.5 mg protein/mL, were co-incubated for 2 h at 37°C and remaining HDL-associated FC was determined. FC transfer was expressed as pmol FC transferred to LDL.

Results:
Within-day and day-to-day CVs for FC transfer to LDL were 2.5% and 2.0%, and for influx, 4.3% and 10.7%. HDL-mol % FC and nmol FC/mg protein differed among five donors and correlated with the donor LDL-FC content. HDL-FC transfer to LDL and macrophages increased linearly with HDL-mol% FC. HDL-FC transfer to LDL correlated with influx to macrophages (Figure).

Conclusions:
Among putatively normolipidemic blood donors, HDL-FC content varies. This is associated with similar variations in HDL-mol% FC, FC transfer to LDL and FC influx to cells. HDL-FC transfer to LDL is a good surrogate of macrophage influx. These assays will be used to determine if a high HDL-FCBI correlates with human ASCVD.
Abstract 1056: Overexpression Of Lysosomal Acid Lipase
Authors: Besler Katrina , Baht Vittoria , Xiang Pinhao , Allahverdian Sima , Ling Elaine , Chan Teddy , Francis Gordon A
Published: July 2024

Background:
Previous evidence suggests arterial smooth muscle cells (SMCs) comprise the majority of atherosclerotic lesion foam cells in human and mouse atheromas, have reduced expression of ABCA1 and lysosomal acid lipase (LAL) compared to macrophages, and retain cholesteryl esters in lysosomes rather than in the cytoplasm as in macrophage foam cells. Exogenous LAL can increase cholesterol efflux from SMC foam cells despite low ABCA1.

Hypothesis:
Overexpression of LAL increases cholesterol efflux from SMC foam cells by ABCA1-dependent and -independent mechanisms.

Methods:
Human vascular SMCs were transfected with 500 MOI AdV-
LIPA
or AdV-
mCherry
(adenovirus) control for 6 h and cholesterol loaded using 100 μg/ml aggregated LDL (agLDL) for 24 h. LAL activity was measured by specific enzymatic assay and location of stored cholesteryl esters (CE) was determined using LAMP-1 (lysosome marker) and BODIPY (neutral lipid) staining by confocal microscopy. Cholesterol efflux to 10 μg/ml apoliprotein A-I (ApoA-I) or 100 μg/ml HDL was determined following ABCA1 siRNA treatment or control, with cholesterol mass determined using Amplex Red assay. Student’s t test was used for statistical analysis of LAL activity, and Mann-Whitney
U
Test for analysis of colocalization of lipid pools.

Results:
LAL activity increased 1.67-fold 72 h after transfection with AdV-
LIPA
compared to SMCs transfected with AdV-
mCherry
(n=5,
p
= 0.001). Increased LAL expression in the absence of cholesterol acceptor resulted in a shift in CE storage from lysosomes to non-lysosomal compartments (Pearson correlation coefficient for BODIPY/LAMP-1 colocalization 0.27 in AdV-
LIPA
vs. 0.47 in AdV-
mCherry
SMCs, n=103,
p
< 0.00001). Preliminary cholesterol efflux studies indicate SMCs overexpressing LAL release cholesterol by both ABCA1-dependent and -independent mechanisms.

Conclusion:
Our study provides evidence that despite low ABCA1 and LAL expression, SMC foam cells mobilize lysosomal cholesterol for cellular trafficking and efflux following supplementation with LAL. Further studies will examine changes in gene expression and the mechanism of ABCA1-independent efflux following LAL supplementation in SMC foam cells.
Abstract 1064: Creation Of Lymphatic Endothelial Cells From Induced Pluripotent Stem Cells For Use In Regenerative Therapy For Lymphedema
Authors: Owsiany Katherine , Carr Sean , Tu Michael , Hekman Katherine
Published: July 2024

Background/Objective:
Lymphedema is a condition characterized by impaired flow of interstitial fluid through the lymphatic system and chronic obliteration of distal lymphatic collecting vessels. Despite advances in our knowledge of the development of lymphatic vessels, there has been little progress translating this knowledge into clinically relevant regenerative therapies. Although lymphatic endothelial cells (LECs) share a lineage with arterial and venous endothelial cell progenitors, currently described processes to derive LECs from stem cells in vitro take up to a month, require embryoid body formation, and have been minimally characterized.

Methods:
We have developed a protocol for the generation of lymphatic endothelial cells from induced pluripotent stem cells (iPSCs) over 11 days. First, iPSCs are directed to become endothelial cell progenitors over 6 days and purified with CD144 magnetic beads. Then, ECs are directed to differentiate into LECs using a 5-day treatment with 10 ng/mL Ang1 alone or in combination with 200 ng/mL VEGFc. Human primary LECs were used as a positive control and iPSC-ECs without Ang1 or VEGFc treatment as well as fibroblasts were negative controls.

Results:
Treating iPSC-derived endothelial cells for 5 days with Ang1 +/- VEGFc yields iPSC-LECs with morphological similarity to primary lymphatic controls and expression of multiple lymphatic markers including Podoplanin, Lyve1, Prox1, and VEGFR3, verified by immunofluorescent staining, western blotting, and flow cytometry across two independent cell lines. In flow cytometry studies, 75% of LECs treated with Ang1 and 72% of LECs treated with Ang1 and VEGFc co-express Podoplanin and Lyve1. There was more than a three-fold increase compared to CD144+ endothelial progenitors (20.4%, p<0.001) while fibroblast expression was negligible (0.6%, p<0.001). Finally, the overall transcriptome of LECs treated with Ang1 was compared to primary human lymphatic cells using single cell RNA sequencing.

Conclusions:
Treatment with Ang1 with or without VEGFc allows fast and efficient generation of LECs from iPSCs. This allows LEC differentiation in vitro from a wide variety of patient specific cell types and improves the practicality of regenerative therapy for lymphedema.
Abstract 1065: Age Increases Platelet Aggregation And Alters The Platelet Transcriptome
Authors: Akinlonu Adedoyin , Muller Matthew , Xia Yuhe , Jindal Manila , Luttrell-Williams Elliot , Goldberg Ira J , Ruggles Kelly , Barrett Tessa , Berger Jeffrey S
Published: July 2024

Introduction:
Age is the strongest risk factor for cardiovascular diseases (CVD). Platelets contribute to CVD pathogenesis and may mediate, in part, the association between age and CVD. In this study, we investigated the impact of aging on platelet activity responses and the platelet transcriptome.

Hypothesis:
Increasing age is associated with increased platelet aggregation and altered transcriptome.

Method:
Adults ≥ 18 years without CVD, off aspirin or NSAIDS were stratified by age tertiles. Platelet activity was measured by flow cytometry and light transmission aggregometry in response to submaximal agonist stimulation with ADP, AA, collagen, and epinephrine. The association between age and platelet variables were adjusted for sex, race/ethnicity, BMI, hypertension, and diabetes. Differentially expressed transcripts by age were identified using the limma package, adjusted for sex and race/ethnicity.

Results:
Among 151 participants, median age was 52 years (IQR 38 to 61), 87 (58%) were female, 75 (50%) were non-White, and 27 (18%) were Hispanic. There was no association between platelet count and size with age. There was a significant association between platelet aggregation in response to multiple agonists and age in both unadjusted and adjusted analyses (
Table
). Using a general linear model, we identified 199 platelet transcripts significantly affected by age (P <0.01, 119-positively, 80-negatively). Gene set enrichment analysis demonstrated upregulation of platelet activity pathways (platelet alpha granule [NES = 2.09, q = 0.0008], platelet activation [NES = 1.90, q = 0.002), and platelet aggregation [NES = 1.77, q = 0.03]) with increasing age.

Conclusion:
Age is independently associated with platelet aggregation in response to submaximal agonist stimulation, a finding supported by the enrichment of platelet activation pathways in the transcriptome. Future studies are needed to elucidate the mechanisms by which age increases platelet aggregation.
Abstract 1071: TGFβ Signaling Negatively Regulates Cardiac Lymphangiogenesis Following Myocardial Ischemia
Authors: Delgado Esteban , Do Long , Bkhache Meriem , Oza Manisha , Liu Xiaolei
Published: July 2024

Cardiovascular disease is the leading cause of death worldwide. Despite high prevalence, standard treatments have made little progress, and developing novel therapeutic treatments to improve cardiac diseases is the top priority of the field. Lymphatics are essential in maintaining tissue-fluid homeostasis, clearing cell debris, and trafficking immune cells. Over recent years, cardiac lymphatics have emerged as a potential target for treatment of cardiovascular diseases, as therapeutic lymphangiogenesis is shown to improve cardiac function following myocardial infarction (MI). However, pathologic lymphangiogenesis induced by MI is reported to be dysfunctional, suggesting cellular or molecular changes occurring in MI directly impact the structure and function of cardiac lymphatics. However, the underlying mechanisms leading to these changes have not been fully explored. In this study we propose TGFβ, a profibrotic signaling molecule increased in ischemic heart diseases, negatively regulates cardiac lymphangiogenesis and lymphatic function following MI. We generated mice with conditional deletion of TGFβ receptor in Lymphatic endothelial cells (LECs) and found that these conditional null (LEC-DKO) mice showed improved post-MI cardiac function and increased lymphangiogenesis compared to controls. Mechanistically, we found that LECs with TGFβ treatment showed increased Endothelial to Mesenchymal Transition (EndMT), and impaired junction integrity. Bulk RNAseq results confirmed these findings as genes involved in EndMT were highly upregulated, and LEC marker related genes were significantly downregulated in LECs in the presence of TGFβ. More importantly, GSEA revealed metabolic reprogramming in LECs treated with TGFβ, with significantly increased expression in glycolysis related genes, but significant downregulation of genes related to fatty acid oxidation. These data provide a possible mechanism by which TGFβ signaling induces LEC metabolic switch to impede lymphangiogenesis and promote cardiac dysfunction following MI, and inhibition of TGFβ signaling in LECs could provide a novel therapeutic strategy to improve lymphatic vascular integrity and lymphangiogenesis, thereby improving cardiac function following MI.
Abstract 1087: Cardiovascular Risk Prediction By Platelet Transcriptomics
Authors: Barrett Tessa J , Cornwell Macintosh , Muller Matthew , Xia Yuhe , Smilowitz Nathaniel , Schlamp Florencia , Newman Jonathan D , Rockman Caron , Voora Deepak , Hochman Judith , Ruggles Kelly , Berger Jeffrey
Published: July 2024

Background:
Hyperreactive platelets are central to atherothrombotic events, including myocardial infarction (MI) and stroke. Currently, no routine clinical test can identify individuals with hyperreactive platelets. The development of a diagnostic platelet hyperreactivity test could identify patients most likely to benefit from antiplatelet therapy to reduce cardiovascular (CV) events.

Aim:
To develop a diagnostic tool to discriminate platelet hyperreactivity and CV risk.

Methods:
Platelet aggregation (0.4 μM epinephrine) was assessed by light transmission aggregometry (LTA) and responses >60% were classified as “hyperreactive” and <40% “normoreactive”. Simultaneously, platelet RNA was collected and sequenced (RNA-seq; n=88). Using machine-based learning, a hyperreactive platelet transcriptomic signature was identified, and a Platelet Reactivity ExpreSsion Score (PRESS) was assigned to each participant. Model training performance was evaluated by 10-fold cross-validation. Signature validation was performed in (a) control cohort without CV disease (n=35), and the diagnostic capacity of PRESS was assessed in patients (b) undergoing coronary angiography (n=28) and (c) lower extremity revascularization (LER, n=129).

Results:
PRESS successfully identified a hyperreactive platelet phenotype in patients with CV disease (AUC 0.81, 95% CI 0.68 - 0.94) and in an independent cohort of healthy participants off antiplatelet therapy (AUC 0.77, 95% CI 0.75 - 0.79). In patients undergoing diagnostic cardiac catheterization, PRESS was significantly higher in patients experiencing an acute MI (β=1.8, adj p=0.02). Among patients undergoing LER, those with a PRESS above the median were more likely to develop a future CV event (adj HR 1.90, CI 1.07 - 3.36, p=0.027).

Conclusions:
A platelet-derived transcriptomic signature identified individuals with hyperreactive platelets and was associated with increased CV risk. A diagnostic platelet reactivity score may facilitate a personalized approach to antithrombotic therapy for CV risk reduction.
Abstract 115: Scavenger Receptor-BI Mediates Endothelial Uptake Of Lipoprotein(a)
Authors: Cabodevilla Ainara G , Bah Alimatou , Younis Waqas , Hussain M M , Goldberg Ira J , Anaganti Narashima
Published: July 2024

Elevated levels of lipoprotein(a) [Lp(a)], an apoB100-containing lipoprotein, are an independent risk factor for atherosclerotic cardiovascular disease, even in the setting of effective reduction of plasma low-density lipoprotein (LDL) cholesterol. To date, the mechanisms whereby Lp(a) exerts its atherogenic effects remain poorly understood, and no treatments aimed at lowering Lp(a) have been approved. Two receptors - scavenger receptor-BI (SR-BI) and activin receptor-like kinase 1 (ALK1) - have been proposed to bind apoB100 and mediate the atherogenic uptake and transcytosis of LDL across the endothelial cell (EC) barrier. EC-specific SR-BI knockout reduces atherosclerosis in both apoE and LDLR knockout mice. We have shown that SR-BI also mediates EC uptake of nascent chylomicrons. Although binding of ALK1 to apoB is believed to occur through the N-terminus of apoB, the receptor is not involved in the uptake of chylomicrons. We assessed which if any of these receptors are involved in Lp(a) uptake by ECs. Despite the presence of apoB100, EC uptake of Lp(a), like apoB48 chylomicrons, was reduced by SR-BI, but not ALK1, knockdown. We then used a series of small N terminal apoB peptides (apoB3-apoB18, with sequences comprising the 3%, 8%, 12%, 15%, and 18% N terminus of apoB) to compete with Lp(a) uptake. Both chylomicron and Lp(a) uptake was inhibited by apoB15 and apoB18, whereas LDL uptake was additionally inhibited by apoB12. These observations suggest that a) the binding sequence to SR-BI is between residues 547-684 of apoB, and b) an additional sequence between residues 365-547 participates in apoB binding to ALK1. Our findings suggest that apo(a) alters the structure of apoB to make it a better ligand for SR-BI uptake by ECs. Because this uptake pathway leads to transcytosis, we hypothesize that the greater atherogenicity of Lp(a) is mediated by its increased binding to and uptake by ECs.
Abstract 154: Hierarchical Redundancy And Contextual Roles Of Vesicle-associated Membrane Proteins (VAMPs) In Platelet Function
Authors: Joshi Smita , Smith Alexis , Alfar Hammodah R , prakhya Kanakanagavalli Shravani , Chanzu Harry , Whiteheart Sidney W
Published: July 2024

Background:
Platelet granule secretion, driven by the SNARE complex involving Vesicle-Associated Membrane Proteins (VAMPs) and t-SNAREs, is essential for vascular integrity. We dissected individual and combinatorial contributions of all four platelet VAMPs to granule secretion, hemostasis, and intracellular cargo trafficking.

Research Questions:
1. Are specific VAMPs critical for granule fusion, or does total VAMP quantity (mass action) dictate secretion? 2. How do differential VAMP deficiencies impact hemostasis in arterial and venous microenvironments? 3. Do VAMPs influence platelet cargo loading and gene expression?

Goals:
Investigate individual and combinatorial roles of VAMPs in platelet biology and their context dependent contributions to hemostasis.

Approach:
We generated V7
-/-
8
-/-
and V2
Δ
3
Δ
7
-/-
8
-/-
mice, assessing granule secretion (dense, alpha, lysosomal) via ATP, PF4/P-selectin, and β-hexosaminidase/LAMP1 release assays, respectively. Hemostasis in arterial and venous models was evaluated with tail bleeding, FeCl3, and venous puncture injury assays. Cargo levels were analyzed using proteomic arrays, and platelet RNA sequencing identified transcriptional changes.

Results:
Global V7/V8 deletion modestly reduced secretion compared to isolated V8 deficiency. Surprisingly, while platelet-specific V2
Δ
3
Δ
7
-/-
8
-/-
exhibited the most dramatic decrease, complete abolition was absent (p<0.01). Alpha and lysosomal secretion were more affected than densegranule release. Both knockout models bled profusely and failed to form stable arterial thrombi (p<0.01). Notably, only V2
Δ
3
Δ
7
-/-
8
-/-
mice displayed significantly prolonged venous bleeding (p<0.01). RNA sequencing revealed >800 upregulated and >500 downregulated transcripts in V7
-
/-
8
-/-
and V2
Δ
3
Δ
8
-/-
platelets, suggesting shared transcriptional control by V7 and secondary VAMPs (V2 and V3).

Conclusions:
VAMP functions exhibit hierarchical redundancy, with V8 presence minimally impacting dense granule secretion and arterial hemostasis. Alpha granule release is more sensitive to VAMP loss. The requirements for platelet granule secretion for hemostasis are context dependent. The deletion of VAMPs disrupts platelet cargo trafficking and transcription.
Abstract 2028: CEPT1 Overexpression Promotes Endothelial Cell Function Through Pparα And Vegf-a
Authors: Khan Tariq , Meade Rodrigo , Belaygorod Larisa , Hafezi Shahab , Semenkovich Clay F , Zayed Mohamed A
Published: July 2024

Objectives:
CEPT1, choline/ethanolamine phosphotransferase 1, is an important mediator of de novo phospholipogenesis and is essential for endothelial cell (EC) survival. CEPT1 catalyzes synthesis of phospholipids that activate the transcription factor peroxisome proliferator-activated receptor α (PPARα). We observed that CEPT1 is elevated in the endothelium of hypoxic and diseased human peripheral arterial segments. Therefore, we hypothesized that
Cept1
overexpression impacts EC angiogenic function and PPARα-mediated signaling.

Methods:
We engineered a
Cre
-induced conditional, EC-specific,
Cept1
overexpression model on a C57BL/6J background. Aortic rings, as well as liver and lung ECs were isolated from

Cept1
fl/fl
Cre

-

and

Cept1
fl/fl
Cre
+

mice (n=3). Aortic rings were implanted in growth-factor reduced Matrigel and microvascular sprouts were evaluated 6 days after implantation.
In vitro
EC proliferation and migration assay were performed. EC gene and protein expression profiles of
Pppara
and
Vegfa
, were evaluated using RT-PCR and Western blot. All experimental conditions were performed in triplicate and were analyzed using the Student’s t-test.

Results:

Cept1
fl/fl
Cre
+

ECs showed significantly increased proliferation and migration (p<0.05; Fig A, B and F). Similarly,

Cept1
fl/fl
Cre
+

aortic rings demonstrated significantly higher microvascular sprouts (p<0.05; Fig C). Gene and protein expression profiling demonstrated that

Cept1
fl/fl
Cre
+

ECs had significantly increased
Vegfa
gene expression (p<0.05; Fig D and F). Western blot demonstrated 79% increase in VEGF-A in ECs of

Cept1
fl/fl
Cre
+

mice. In HUVECs overexpressing
Cept1
,
Ppara
siRNA decreased EC migration (p<0.05; Fig E).

Conclusion:
Cept1
pro-angiogenic signaling appears to be
Ppara
-dependent and mediated by VEGF-A. CEPT1 content in diseased peripheral arterial segments may represent a potential compensatory mechanism for enhanced recovery during chronic limb ischemia.
Abstract 2040: Rapamycin Extends Life Of Matrix Gla Protein Knockout Mice Independent Of Vascular Calcification Content.
Authors: Behzadi Parya , Wendling Andrew A , Chu Claire , Crane Alex , Moorehead William J , Wong Ryan , Brown Mark J , Suresh Swathi , Cuevas Rolando A , St. Hilaire Cynthia
Published: July 2024

Background:
Peripheral artery disease (PAD) is characterized by the narrowing and blockage of the vessels in lower extremities. While atherosclerosis has been the assumed culprit of PAD recent data now shows that medial arterial calcification (MAC) is the cause of below-the knee ischemia. MAC promotes thrombosis via inward remodeling, reduced elasticity, and arterial stiffness. The mechanisms driving MAC pathogenesis are unclear. Matrix Gla Protein (MGP) is an inhibitor of vascular calcification and present in arterial vessel walls. MGP
-/-
mice exhibit extensive MAC throughout their vasculature and die within 5-7 weeks due to aortic rupture or heart failure. The drug rapamycin functions as an inhibitor of cell proliferation and an activator of autophagy.

Hypothesis:
That rapamycin reduces MAC in MGP
-/-
mice via inhibiting inward remodeling and activating autophagy.

Methods:
MGP
+/+
and MGP
-/-
mice treated with vehicle or rapamycin (5mg/kg, 3x weekly) starting at 10 days old for 2 weeks. Calcification was quantified via micro-computed tomography (uCT). Collagen structure and elastin quality were assessed histological staining. Markers of proliferation (Ki-67), cell death (Caspase 3), autophagy (LC3), and calcification (Runx2) were quantified via immunofluorescence staining. Western blot was used to assess autophagy flux and proliferation in smooth muscle cells (SMCs) isolated from MGP
+/+
and MGP
-/-
mice.

Results:
While rapamycin treatment doubled the life-expectancy of MGP
-/-
mice, the calcification content was unchanged compared to the vehicle. However, rapamycin caused a significant decline in mineral density. Higher collagen content and improved elastin structure were observed in MGP
-/-
treated with rapamycin compared to vehicle. Runx2 and Ki-67 were elevated in MGP
-/-
mice compared to MGP
+/+
, but there was no difference in response to rapamycin treatment. No difference was observed in caspase 3 levels. Under rapamycin treatment, MGP
-/-
SMCs exhibited higher autophagy flux than MGP
+/+
SMCs.

Conclusion:
While rapamycin did not reduce MAC, our data suggest that rapamycin potentially preserves the integrity of the aortic vessel wall by inducing autophagy, which requires further investigation.
Abstract 2064: Identifying A Novel Protein Protecting Against Atherosclerosis And Vascular Dysfunction
Authors: Hussein Hebaallaha , Balakrishnan Bijinu , Basri Rabia , Becker Chloe , Sharma Vishva , Malgor Ramiro , Khan Mahmood , GOKCE Noyan , Puri Vishwajeet
Published: July 2024

Atherosclerosis and ischemic heart disease are the leading causes of death in the US. The onset of atherosclerosis begins with endothelial dysfunction and ultimately progresses to occlusive plaques by mechanisms that are still incompletely understood. The basic mechanisms of atherosclerosis involve lipid accumulation and immune activation in the vascular wall. These processes are highly regulated, poorly understood, and lead to inflammation and atherosclerotic plaques in the arterial wall. We recently identified that Fat-Specific Protein 27 (FSP27) plays an important role in regulating vascular function. Using both clinical approaches that involved human biospecimens and “humanized” murine models of endothelium-specific FSP27-overexpressing transgenic (E-FSP27tg) mice, we demonstrated that FSP27 protects against high-fat diet (HFD)-induced dyslipidemia, endothelial dysfunction, and vasomotor impairment. We next hypothesized that FSP27 plays a critical role in protecting against atherosclerosis primarily via its action on endothelial cells. To test our hypothesis, we generated an innovative mouse model expressing the human FSP27 transgene specifically in the vascular endothelial cells of atherosclerosis-prone Apoe
-/-
mice, Apoe
-/-
-E-FSP27tg. Our preliminary data show that high-fat-high-cholesterol fed Apoe
-/-
-E-FSP27tg mice are strongly protected against plaque formation in numerous vascular territories including carotids and aorta upon feeding high-fat, high-cholesterol diets. Mechanistically, we found that FSP27 protects against oxidative stress and inflammation in the endothelial cells. Overall, our study discovered the novel beneficial cardiovascular effects of FSP27 where it protects against the development of vascular dysfunction, inflammation, and atherosclerosis.
Abstract 2066: Relationship Between Stat3 And Endoplasmic Reticulum Stress Induces Lipopolysaccharide-induced Endothelial To Mesenchymal Transition In Vascular Endothelial Cell
Authors: Jin Yujin , Heo Kyung-Sun
Published: July 2024

Background:
Endothelial to mesenchymal transition (EndMT) is a process that involves the loss of endothelial cell (EC) phenotype and induced plaque instability during the progression of atherosclerosis. Primarily, EC injury, caused by an accelerated inflammatory signaling pathway, plays a crucial role in activating EC dysfunction by inducing endoplasmic reticulum (ER) stress and disrupting EC junctions. In the present study, we investigated the molecular mechanisms between EndMT and ER stress on STAT3-mdeiated inflammation under LPS-treated human umbilical vein EC (HUVEC).

Methods:
To explore the EndMT progression under chronic inflammation, LPS-treated HUVEC were used. Western blot, siRNA transfection, and immunofluorescence assay were employed to evaluate signaling pathways and target proteins. The phenotypic switching of ECs on the endothelium of the mouse aorta after challenging 10 mg/kg LPS was assessed by
en face
staining.

Results:
Deactivation of STAT3 by stattic pretreatment abolished not only LPS-mediated STAT3 phosphorylation, but also VCAM-1 and ICAM-1. In addition, STAT3 inhibition surprisingly reversed LPS-induced mesenchymal markers, including SM22α and vimentin, and the suppression of endothelial marker VE-cadherin. Mechanistically, stattic treatment suppressed LPS-induced EndMT progression by inhibiting protein expression and nuclear translocation of Smad2/3. Furthermore, LPS-induced chronic inflammation disrupted EC homeostasis by PERK or IRE-1α/CHOP-mediated ER stress, whereas stattic abolished LPS-mediated PERK and CHOP expression. Interestingly, depletion of CHOP by siRNA transfection significantly abolished LPS-mediated EndMT progression (p<0.001. Figure 1). Notably, the absence of CHOP significantly suppressed phosphorylation of STAT3 and its nuclear expression as well. Consistent to in vitro data, STAT3 inhibition by stattic injection significantly suppressed LPS-mediated STAT3 activation as well as CHOP expression in the mouse endothelium.

Conclusion:
Collectively, the STAT3-CHOP axis may contribute to the loss of EC function and induce mesenchymal differentiation by aggravating chronic inflammation and EndMT.
Abstract 2084: Endothelial Cells With Silenced Flrt2 Expression Exhibit Enhanced Retention On Vascular Graft Materials Under Fluid Shear Stress In Vitro
Authors: Rao Shashanka , Wolfe Jayne , Chen Yiliang , Tefft Brandon J
Published: July 2024

Myocardial infraction is a leading cause of death, and surgical intervention is often required to treat severe coronary artery disease. Autologous vessel grafts are the only option for bypass graft procedures. Synthetic vascular grafts less than 6 mm diameter fail due to unacceptable patency rates (~60% at 1 year). The loss of patency is accredited to platelet activation, thrombosis, and neointimal hyperplasia. A promising strategy to improve patency is to endothelialize synthetic vascular grafts ex-vivo, but previous attempts have met with modest success due to detachment of cells upon exposure to fluid shear stress. We previously used RNA-seq to characterized gene expression of endothelial cells (ECs) which remained adherent to synthetic grafts post shear stress. We found that fibronectin leucine-rich transmembrane protein 2 (FLRT2) was significantly downregulated in adherent subpopulations. We therefore hypothesize that suppressing FLRT2 expression in ECs will result in better retention on synthetic vascular grafts upon exposure to shear stress. We used silencing RNA approach using lipofectamine and confirmed successful knockdown of FLRT2 expression (Si-FLRT2). We then seeded Si-FLRT2 cells onto a commercially available Goretex graft material and subjected them to fluid shear stress of for 20 min. We then stained the samples with DAPI. We observed increased retention of Si-FLRT2 ECs on Goretex graft material in comparison to non-specifically targeted scramble siRNA containing ECs. In addition, we compared both vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) levels to assess endothelial cell activation and found that silencing of FLRT2 lead to decreased levels of both VCAM-1 (by 49.2%±3.5%) and ICAM-1 (by 57.1%±3.4%) suggestive of decreased endothelial cell activation. In conclusion, we show that FLRT2 plays an important role in adhesion of ECs onto a commercially available synthetic graft material and silencing its expression ex-vivo may have beneficial clinical outcomes. In addition, we found that FLRT2 silencing may lead to decreased endothelial cell activation and we would like to further investigate the molecular mechanisms underlying the role of FLRT2 in both EC adhesion and activation.
Abstract 2121: SUV39H1 Mediated Regulation Of KLF4 And KDM4a Coordinate Smooth Muscle Cell Phenotypic Plasticity
Authors: Chatterjee Payel , Chakraborty Raja , Sizer Ashley , Xie Yi , Hwa John , Martin Kathleen A
Published: July 2024

Background:
The phenotypic plasticity of vascular smooth muscle cells is central to growth and remodeling processes, but also underlies many cardiovascular pathologies, including atherosclerosis, restenosis, and intimal hyperplasia. This ability of VSMCs to reversibly differentiate and dedifferentiate is incompletely understood. SUV39H1, a histone methyltransferase, that generates the H3K9Me3 mark results in epigenetic transcriptional repression. We hypothesized that SUV39H1 plays a key role in VSMC plasticity.

Methods:
We applied knockdown, qPCR, western blotting, chromatin immunoprecipitation (ChIP) assays, RNA-Seq, and murine vascular injury to determine the role of SUV39H1 in VSMC plasticity.

Results:
A qPCR array screen of epigenetic regulators revealed that SUV39H1 is upregulated with PDGF-induced dedifferentiation but downregulated with rapamycin-induced differentiation in hCASMCs. Knockdown of SUV39H1 induces differentiation-specific hCASMC contractile genes and functional contractility while decreased hCASMC migration, proliferation, and de-differentiation-associated gene expression. RNA-seq transcriptomics confirmed changes in multiple pathways consistent with a role for SUV39H1 in promoting hCASMC dedifferentiation. Mechanistically, SUV39H1 knockdown suppressed expression of KLF4, a master dedifferentiation factor, decreasing KLF4 mRNA stability and upregulating miRNA143. SUV39H1 knockdown also increased expression of KDM4a, an enzyme that removes H3K9me3 marks.
In vivo,
we noted a significant increase in SUV39H1 and H3K9me3 expression in murine carotid artery ligation-induced intimal hyperplasia compared to uninjured vessels.

Conclusion:
We identify SUV39H1 as an epigenetic regulator of VSMC phenotype whose expression and activity increase with dedifferentiation
in vitro
and
in vivo
. PDGF promotes H3K9me3 repressive marks at contractile genes by promoting expression of SUV39H1 (writer), which also inhibits the KDM4a (eraser) and SUV39H1 also regulates KLF4 expression. Understanding the role of SUV39H1 in VSMC plasticity may reveal new therapeutic strategies for treating vascular diseases.
Abstract 2146: Vascular Loss Of Type VIII Collagen Improves Venous Remodeling After Arteriovenous Fistula Creation In Mice
Authors: Labissiere Xochilt , Rojas Miguel G , Martinez Laisel , Vazquez-Padron Roberto
Published: July 2024

Objective:
Collagen VIII is a basement collagen upregulated in processes of vascular remodeling. We recently demonstrated the association of increased
COL8A1
expression and arteriovenous fistula failure in a human cohort. This study analyzed the effect of loss of collagen VIII in a murine arteriovenous fistula model.

Methods:
Arteriovenous fistulas were created between the external jugular vein and common carotid artery of 5/6 nephrectomized
Col8a1/a2
null mice and wild type controls (n=8-9 per group). Intimal hyperplasia was quantified in Masson’s trichrome stained sections. Inflammation was evaluated by immunofluorescence using anti-galectin 3 (Mac2) antibodies and expressed as % positive cells/total cells.

Results:
AVF cross-sections from collagen VIII null mice demonstrated a significant reduction in intimal hyperplasia compared to controls (7.6 vs 107.8, p = 0.004). AVF from collagen VIII null mice also presented a significantly lower proportion of infiltrated macrophages compared to fistulas in wild type mice (1.00 vs. 6.05%, p=0.004).

Conclusions:
This study shows for the first time that absence of collagen VIII improves postoperative remodeling of murine AVF. Importantly, our findings suggest a dual role of collagen VIII in vascular inflammation and wall remodeling.
Abstract 2147: Impact Of Donor Age On Microvasculature Self-assembly Of Induced Pluripotent Stem Cell Derived Endothelial Progenitors
Authors: Larsen Bryce , Faulkner Sara , Zoldan Janet
Published: July 2024

Introduction:
Current vascular tissue-engineering using induced pluripotent stem cells (iPSCs) predominantly relies on somatic cells obtained from newborn or fetal donors. However, iPSCs sourced from aged individuals exhibit epigenetic differences in angiogenic genes that may hinder their utility in iPSC derived vasculature. We hypothesized that, due to these differences, iPSCs from aged donors will exhibit decreased vasculogenic potential.

Methods:
We differentiated iPSCs into endothelial progenitors (iEPs) and encapsulated them in an interpenetrating polymer network hydrogel containing norbornene-functionalized hyaluronic acid and Collagen I at cell densities and hydrogel stiffness optimal for vasculogenesis. Seven days after encapsulation, the hydrogels were fixed, and the resulting microvasculature imaged. We then utilized a previously developed computational pipeline to quantify several vasculogenic properties.

Results:
We derived iEPs from four separate iPSC lines: 2 sourced from neonatal donors (ND) and 2 from mature donors (MD). Irrespective of donor age, differentiation yielded ~15% (n=4) iEPs of the total cell population. The two ND lines developed a similar plexus, whereas this structure was diminished in one adult line and absent in the second. Between iEPs matched for sex and somatic cell origin, there is a 50% decrease in vessel volume fraction and number of links between vessels, a 66% reduction in the percentage of vessels connected to the largest vessel, and a 43% reduction in branch points between vessels (
Fig. 1
).

Conclusion:
Overall, these data suggest MD iEPs are less likely to form dense interconnected microvasculature as compared to ND iEPs. Interestingly, the average diameter did not change across all cell lines. This suggests that while MD iEPs exhibited cell clustering, they were less likely to branch out and interconnect with other clusters of iPSC-EPs to form interconnected networks