bluuebunny/crossref_metadata_2025 huggingface dataset:
Pseudo Identity in Cassava Rice as A Traditional Religious Identity Struggle Against Contemporary Religious Discrimination
Authors: Ardiansyah Anton , Sihabudin Ahmad , Dua Mikhael
Published: July 2024

The people of Cireundeu Village are known as a community group that still maintains the tradition of ancestral heritage of consuming rasi (cassava rice) as a staple food since 1918. This consumption process imbues rasi with the meaning that it is part of their culture.
This study employed a qualitative approach with a semiotic method in decoding the meaning of rasi among the people of Cireundeu indigenous village.
The purpose of this study was to find out 1) what meaning is constructed from rasi as their staple food 2) and what identity is built among the Cireundeu indigenous people.
The results of this study show that 1) rasi is a form of struggle for religious identity that has been discriminated against, namely the struggle for belief in Sunda Wiwitan as their religious identity and 2) rasi as a group identity is a pseudo-identity of the Cireundeu indigenous people who primarily exist to maintain group beliefs in their beliefs.
Cultural Heritage and Tourism: The Case of Dellys in Algeria
Authors: Necissa Yamina , Rayane Moussaoui
Published: July 2024

Cultural heritage is an invaluable resource for the development of cultural tourism. It offers a wide range of economic and social opportunities, contributing to job creation, income generation and the promotion of local identity. However, to fully exploit its potential, heritage needs to be carefully integrated into an overall tourism strategy. Such a strategy must include specific measures for the preservation and enhancement of cultural heritage.
Recognizing the profound synergies between cultural heritage preservation and tourism development strategies, we undertook an in-depth analysis of the current heritage situation in the town of Dellys. This analysis identified the challenges and opportunities associated with heritage enhancement in the context of tourism and local development projects.
The results reveal that the implementation of the current tourism strategy in Dellys is not fully achieving its objectives. Nevertheless, its effective realization, coupled with better coordination between players in the sector, could make a significant contribution to safeguarding, revitalizing and enhancing the town's heritage.
Reinventing Urban Agriculture in Algerian Urban Policies: Stakeholders' Visions and Operational Strategy
Authors: Korchi Meriem , Bendjaballah Ouassila
Published: July 2024

Climate change and rapid urban growth are driving an increased demand for global resources, exacerbating environmental challenges. Addressing these issues necessitates the implementation of diverse strategies across various scales. Urban agriculture emerges as a potential solution to foster the resilience of urban entities. However, in the Mediterranean region, notably in Algeria, urban agriculture remains inadequately addressed in public policies, with agricultural plots in cities often earmarked as land reserves for future urban development.
This study aimed to assess the perspective of urban actors in Annaba regarding urban agriculture and to identify and map agricultural activities within the city, with the goal of integrating them into urban policy frameworks. Interviews were conducted with decision-makers and farmers, complemented by spatial analysis.
Findings revealed that although there is interest in urban agriculture, it is not effectively integrated into institutional and legal frameworks. Despite modest agricultural production primarily for self-consumption, urban agriculture projects hold potential for adoption in Algerian cities, offering a pathway towards enhancing urban sustainability.
Analysis of Calculation of Cost of Production Using the Full Costing Method
Authors: Ulum Bustanul , Maulidayanti Diah Wulan , Rusdiyanto Rusdiyanto
Published: July 2024

The goal of the study is to investigate and demonstrate that, for Micro, Small, and Medium-Sized Businesses, applying the complete costing method for calculating manufacturing costs results in a more accurate computation. This study is crucial for entrepreneurs since it helps them determine the precise cost of manufacturing things and determines how much profit they should make. This study employs qualitative descriptive methods. The results of this study's comprehensive costing technique calculation of manufacturing costs for commodities are more accurate for use in micro, small, and medium-sized enterprises.
Exploring the Distinctive Symbolic Interplay: Modern Vespa Piaggio User Community Dynamics in Makassar, Indonesia
Authors: Abidin M. Zainul , Muhammad Rahmat , Syam Ridwan
Published: July 2024

Symbolic interaction is a form of reciprocal action carried out by two or more people to be able to translate and understand the symbols used in interaction. This includes interactions carried out by people who behave Fomo in a community. The research aims to find out the forms of Fomo behavior and understand the meaning of social action and symbolic interaction of members of the Makassar Move Community. The research used a qualitative approach with a case study strategy by determining informants using purposive informant selection. The results of this study found that Move Makassar members have a form of Fomo behavior by being so active in finding and sharing information about vespa matic and Move Makassar activities, always wanting to be involved in every activity, and excessive consumption of vespa matic accessories and modifications. The meaning of social action in the symbolic interaction of Move Makassar members is understood as an action that upholds the values of equality, togetherness and intimacy among them so that a sense of kinship arises.
Sale of Rights in Dispute between Sharia Law and Algerian Law
Authors: Gherbi Soreya , Boudjani Abdelhakim
Published: July 2024

The sales contract is one of the most important contracts in the lives of individuals and nations, as it is the customary means of exchanging money after the decline of bartering. With the passage of time, the shortcomings of bartering became evident due to the development of industry and commerce, and the varying needs of people. This led to the emergence of the sales contract, as it became a more suitable method of transaction compared to bartering, serving as a common ground for transactions between parties more than bartering. The aim of this research is to analyze the concept of rights sale in the conflict between Algerian law and Islamic law, and to understand the differences and similarities between the two legal systems in this context. Methods Used: An analytical methodology was employed, involving a comparative study of laws and judicial rulings in Algerian law and Islamic law pertaining to rights sale. Findings: The study revealed significant differences between Algerian law and Islamic law regarding the concept, conditions, and transactions of rights sale. Additionally, the findings pointed out some of the challenges and contradictions that arise when applying Sharia principles in a non-Islamic legal system.
Communication Strategy for Handling Cases of Stunting and Bad Nutrition Through The 2023 Stunting Free Program (Bessan) in Cipocok District, Serang City
Authors: Azhar Firdausi Indrianti , Aries
Published: July 2024

Cases of stunting and malnutrition that children suffer from or have the potential to have negative consequences for the future of the younger generation. There are 91 cases in Cipocok Jaya District spread across 8 villages. This occurs due to various factors including poverty, low education, lack of knowledge regarding good nutritional intake for children. A strategy is needed to communicate intensively with communities with stunted and malnourished children so that they can get out of this situation. This research uses a qualitative approach through observation and interview data collection. The research results show that the communication strategy in handling stunting and malnutrition through the Stunting Free 2023 (Bessan) program in Cipocok Jaya District, Serang City uses a communication strategy approach which includes getting to know the audience, compiling messages, determining methods and selection and use of media which has been effective. The communication strategy to get to know the audience is carried out through surveys and data collection, identifying indicators of sufferers and potential stunting. The second strategy determines the micro-scale goal of reducing stunting sufferers, the macro-scale of stunting cases at zero, where this treatment accommodates community self-help. The next communication strategy is to develop messages so that people's lifestyle behavior becomes healthy by conveying the importance of nutritious food intake for children. Methods for determining the methods used are socialization and counseling, accommodating community self-help through healthy exercise activities, regular recitations, and Gowes Lur events. This series of activities is a collaborative innovation in Cipocok Jaya District which involves stackholders starting from the Serang City Health Office, Serang City DP3AKB, KUA, as well as cadres from both posyandu cadres and PKK Community Institution cadres. Research findings show the importance of consistent mentoring or accompanying posyandu cadres and PKK community organizations as an important instrument for a sustainable communication strategy in efforts to reduce stunting and malnutrition rates.
The Urgency of Appointing a Notary as The Official for Creating Waqf Pledge Deeds (PPAIW) for Shares Based on Law Number 41 of 2004 on Waqf
Authors: Wahab Abdul , Sambas Nandang , Sumiyati Yeti
Published: July 2024

The object of waqf known to the public up to this point is still synonymous with immovable property (fixed assets). In Indonesia, regulations related to waqf are governed by several laws, including Law No. 41 of 2004 on Waqf. However, there are no specific regulations regarding share waqf. Additionally, concerning the creation of share waqf pledge deeds by a notary as the PPAIW, the regulations on waqf pledges stipulated in Article 27 of the Minister of Religious Affairs Regulation No. 73 of 2013 on Procedures for the Waqf of Immovable and Movable Assets Other than Money state that a notary is appointed as PPAIW by a Ministerial Decree. To be appointed by the Minister as PPAIW, one must meet the requirements of being a Muslim, trustworthy, and certified in waqf competence by the Ministry of Religious Affairs.
These requirements pose a barrier for notaries to become PPAIW because there has been no socialization and implementation of the competence certification requirement. Moreover, the requirement to be Muslim means that not all notaries can become PPAIW. The aim of this study is to understand the implementation of regulations regarding notaries as PPAIW in the creation of Waqf Pledge Deeds. This research is a type of normative juridical research, with a sociological juridical approach. The findings indicate that the regulation and implementation of notaries as PPAIW for shares are not yet clear and firm, necessitating a broader legal review in relation to the principle of the benefits of share waqf for the community.
Legal Certainty in The Validity of Interfaith Marriages Performed Abroad
Authors: Asih Nur , Uwiyono A. , Wulandari Wahyuni Retno
Published: July 2024

This research examines in more depth how legal certainty regarding the validity of marriage certificates of different religions is held abroad after being recorded and reported to the Population and Civil Registry Office (Dukcapil), where the couple resides upon returning to the homeland. With lex loci celebrationis and lex fori principles, the state is expected to accept, recognize, and respect as the basis for recording and registration with the Dukcapil. This type of research is normative research using a statutory and factual approach. The data collection method used literature study, field research, and interviews. Based on the results and discussion of the research, it is found that Indonesian Citizens (WNI) who enter into marriages of different religions abroad under International Civil Law (HPI) are valid and recognized. However, when viewed from the applicable marriage law in Indonesia, it is invalid because it still has to be based on religious law. However, sociologically, the community still recognizes and accepts the existence of interfaith marriages.
Evaluation of antioxidant activity in crude polysaccharide extracts from two date varieties (Tazerzait “Azerza” and Deglet-Nour)
Authors: Belhachemi Mohammed Habib , Belmir Sara , Mebarek Mohammed Oulad , Reffis Mohammed , Achour Fatima Zahra
Published: July 2024

The fruit of the date palm, Phoenix dactylifera L. is extensively consumed in Arabian countries and is esteemed for its nutritional attributes, boasting high carbohydrate content and specific vitamins. This study aims to assess the antioxidant potential of polysaccharide extracts from the date palm fruit (Phoenix dactylifera L.) from two selected cultivars, Tazerzait “Azerza” and Deglet-Nour, originating from the Ghardaia region. Crude polysaccharides were extracted from the dates using distilled water followed by ethanol precipitation. Yields from this process were 3.14% for the Tazerzait cultivar and 4.75% for the Deglet-Nour cultivar, indicative of significant polysaccharide content, known for their diverse health benefits. Antioxidant activity was evaluated in vitro through DPPH and ABTS assays. Polysaccharide extracts exhibited noteworthy antioxidant activity, notably the Tazerzait variety. In the DPPH assay, the Tazerzait extract displayed an IC50 value of 4.66 mg/mL, indicating effective scavenging of free radicals. Moreover, in the ABTS assay, the extract demonstrated even more potent activity, with an IC50 value of 0.49 mg/mL, underscoring its robust antioxidant properties. These results underscore the strong antioxidant potential of Tazerzait polysaccharides. Polysaccharide extracts from the date palm fruit, particularly from the Tazerzait cultivar, exhibit significant antioxidant activity, highlighting their potential as natural antioxidants. This suggests avenues for their utilization in pharmaceutical and food industries to enhance human health.
Bio-treatment of Cheese Whey by Black Soldier Fly Larvae (Hermetia illucens) reared in Algeria
Authors: Hanafi Tahar , Meziane Kaci Zoubida , Megateli Smain , Moussaoui Badreddine , Guemou Laid , Reghioui Bachir
Published: July 2024

Whey, a by-product of cheese industry, poses environmental challenges when discharged untreated. This study aimed to mitigate whey pollution and transform it into value-added products using black soldier fly larvae (BSFL). By investigating whey effects on BSFL growth and performance, an innovative waste management strategy was explored. The five-old-day larvae were fed with four diets; dry spent coffee ground (SCG) as the control diet, while SCG wetted with acid, sweet and fermented whey were the test diets. The results showed that whey types increased significantly (p < 0.05) BSFL growth compared to the control, the dry weight (53.83±0.49 Vs 25.10±0.18 mg/larva), growth rate (1.59±0.06 Vs 0.44±0.01 mg/day) and survival rate (78.83±1.25 Vs 68.50±1.5 %) of larvae and reduced their development time (28.00±01 Vs 37.00±01 day). Furthermore, mixing SCG diet with whey enhanced positively their performance by increasing the waste reduction (26.73±0.81 Vs 16.87±0.37 %), waste reduction index (0.85±0.03 Vs 0.45±0.01 %/day), approximate digestibility (36.49±1.52 Vs 20.29±0.53 %) and bioconversion rate (17.94±0.47 Vs11.65±0.52%). Adding whey to SCG diet increased protein content of BSFL (40.11% vs. 34.18%) and reduced fat (36.06 vs. 37.87 %) in dry mass. Whey also improved the composition of frass by increasing the phosphorus and total nitrogen levels. This approach demonstrates a sustainable solution for dairy industries waste, contributing to the circular economy and offering potential applications in animal feed and fertilizer production.
Entre continuidades e rupturas: a representação do cientista e da ciência a partir de imagens geradas pelo ChatGPT
Authors: Neves Luiz Felipe Fernandes , Medeiros Amanda , Massarani Luisa
Published: July 2024

Imagens estereotipadas do cientista e da ciência têm o potencial de interferir na confiança e em movimentos de aproximação/ afastamento do campo científico por parte da população. Diante do crescente uso da Inteligência Artificial (IA) no contemporâneo, objetivamos analisar imagens geradas pelo ChatGPT sobre “cientista” e “ciência”, buscando observar continuidades e rupturas na manutenção de estereótipos raciais, de gênero e etário, além de estereótipos que apontam para as ciências exatas e biológicas como modelo de referência. De um modo geral, os resultados sugerem a reafirmação do cientista como homem branco, bem como das ciências exatas e biológicas como centrais na sociedade. Indicam ainda mudanças na prática científica, que vai de individual a coletiva, e trazem uma visão otimista quanto à inclusão do jovem cientista, e a facilidade de acesso a recursos hipertecnológicos.
Comunicar e educar num Brasil do “quase”: o impacto da branquitude estruturante
Authors: Prandini Paola Diniz
Published: July 2024

A colonização portuguesa, no Brasil, deixou marcas profundas na população. Uma delas é a hegemonia branca – conceituada como branquitude – que transversaliza os modos de ser e de estar brasileiros (apesar de não se limitar apenas a esta nação). Nesse sentido, este artigo pretende analisar alguns dos impactos causados pela branquitude estruturante no país, bem como apresentar caminhos possíveis para a transformação dessa realidade, por meio da construção coletiva de potenciais educomunidades. Desse modo, espera-se que a leitura crítica do Brasil pós-colonial, ora apresentada, colabore para o estabelecimento de estratégias mais justas de comunicar e de educar, colaborativamente, em território nacional.
Redes do imperialismo na agenda do combate à desinformação
Authors: Albuquerque Afonso
Published: July 2024

In less than a decade, a new paradigm, which argued that the world was experiencing an “information disorder”, took the international academic debate by storm. This study explores the hypothesis that forces outside academia sponsored the emergence of this new paradigm and seeks to investigate what were these forces, how they acted, and what their objectives were. This study argues that the expansion of the information disorder paradigm constitutes a process of intellectual imperialism that mainly aims to provide institutions based in the United States and its allies with the global authority to define what is or is not disinformation.
Transformações sociotécnicas na Comunicação e Educação: do analógico ao digital
Authors: Oliveira Thaiane Moreira de , Castro Gisela , Ricaurte Quijano Paola
Published: July 2024

A introdução da tecnologia na educação tem se tornado uma tendência global, com diversos países e estados adotando medidas para modernizar o ensino e otimizar processos administrativos. Alguns exemplos incluem o uso de Inteligência Artificial na produção de materiais didáticos e na automação de práticas pedagógicas, como avaliações de desempenho e planejamento de atividades. Esse avanço tecnológico tem desencadeado mudanças profundas na educação. Essas mudanças enfrentam desafios significativos, como questões de privacidade de dados, acesso equitativo à tecnologia e necessidade de formação de professores. Além disso, há preocupações sobre a narrativa do “progresso” e sua ligação com dinâmicas econômicas, que podem obscurecer as verdadeiras motivações por trás dessas transformações. O artigo propõe discutir essas transformações sociotécnicas, especialmente diante da crescente plataformização da educação, da expansão do ensino a distância e do desenvolvimento acelerado de ferramentas de IA. Também enfatiza a importância de regulamentações rigorosas e investimentos adequados em infraestrutura para garantir a soberania informacional do país.
Abstract 1004: Three-dimensional Characterization Of Sex Differences In Abdominal Aortic Aneurysm Progression Via Vascular Deformation Mapping
Authors: Braet Drew , Delbono Luciano , Spahlinger Greg , Graham Nathan , Arora Akul , Figueroa C A , Eliason Jonathan , Burris Nicholas S
Published: July 2024

Introduction:
Although Abdominal Aortic Aneurysms (AAA) are more common in men, women are at greater risk for AAA growth and rupture. Vascular deformation mapping (VDM) is a novel technique that utilizes deformable image registration to qualify and quantify three-dimensions (3D) AAA growth using routing clinical computed tomography angiogram (CTA) images.

Hypothesis:
We hypothesize that females will have higher rates of focal AAA growth when compared to males.

Aims:
In this study we aimed to leverage VDM to investigate sex differences in AAA growth.

Methods:
We identified patients with infra-renal AAA and ≥2 CTA from 2010-2021. Patients with non-contrast CT scans or poor image quality were excluded. Propensity score matching was used to identify a cohort of males and females matched on age, hypertension, and smoking history. AAA were segmented and VDM was conducted using a multi-step image registration of aortic CTA studies to generate a deformation field which is then used to quantify 3D AAA growth rate (GR). Patient characteristics, maximum diameter (Dmax), AAA volume (Vmax), and GR were obtained. Statistical shape modeling was utilized to compare average 3D growth patterns between males and females.

Results:
Seventeen male and females with AAA were matched. There was no differences between sexes in race, diabetes, symptomatic status, or repair. Males had a trend toward larger Dmax (4.9 ± 0.6 vs 4.5 ± 0.6 cm, p = 0.055) and had Vmax at baseline (118.7 ± 69.9 vs 76.6 ± 35.0 cm
3
, p = 0.034). There was no difference between sexes when looking at change in Dmax or Vmax. Male and female AAA had no difference in average GR (0.11 ± 0.08 vs 0.17 ± 0.22 cm/year, p = 0.293). Shape models demonstrated more diffuse AAA growth for males and more eccentric AAA growth for females (Figure 1).

Conclusions:
Despite comparable changes in Dmax, Vmax, and GR, women demonstrated a more eccentric distribution of AAA growth, a finding which may contribute to higher rupture risk in women.
Abstract 1008: Phenol-Based Denervation Induces Sustained Abdominal Aortic Remodeling
Authors: Chao Calvin L , Dang Caitlyn , Reddy Nidhi , Alharbi Sara , Doan Jimmy , Karthikeyan Akashraj , Applewhite Brandon , Jiang Bin
Published: July 2024

Introduction:
Aortic remodeling remains an important contributor to the pathogenesis of aortic disease, including occlusive, aneurysmal, and dissection disease states. The paucity of medical therapies has driven considerable interest in elucidating the pathogenesis of these conditions; new therapeutic targets are critically needed. Prior studies have identified sympathetic nervous system signaling (SNS) as a critical regulator of arterial wall homeostasis with potent effects on inflammation and vascular remodeling.

Hypothesis:
This study seeks to evaluate the impact of sustained denervation on morphology and wall homeostasis of the abdominal aorta. We hypothesized that topical phenol denervation of the rat abdominal aorta would induce sustained reduction in nerve fiber density with concomitant pathologic changes to tissue morphology.

Methods:
Male and female Sprague-Dawley rats (n=12), aged 3 months, underwent midline laparotomy for infrarenal aorta exposure. Chemical denervation was induced via a one-time topical application of 10% phenol (n=6), while sham controls received phosphate-buffered saline (n=6). Animals were allowed to recover and subsequently sacrificed after 6 months for analysis encompassing morphology, histology, and immunohistochemistry.

Results:
At 6 months post-treatment, abdominal aortas subjected to phenol denervation exhibited a significant reduction in nerve fiber density compared to sham controls (92.8 ± 20.3 fibers/mm
2
vs 170.4 ± 79.0 fibers/mm
2
, p=0.042). Denervated aortas demonstrated increased elastin fragmentation breakage scores (2.8 ± 1.1 vs 1.1 ± 0.4, p=0.0056). Phenol denervated rats also had significantly decreased expression of vascular smooth muscle proteins α-SMA (32.6 ± 3.9% vs 24.6 ± 3.3%, p=0.0034) and MYH11(14.2 ± 5.7% vs 21.6 ± 3.5%, p=0.023), as well as elevated adventitial microvascular density (58.9 ± 25.2 vessels/mm
2
vs sham control 18.5 ± 8.7 vessels/mm
2
, p=0.0040).

Conclusion:
Single-timepoint phenol-based chemical denervation induces sustained alterations in abdominal aortic morphology and vascular remodeling over a 6-month period. These findings underscore the potential of the SNS as a therapeutic target for aortic pathologies.
Abstract 1010: Novel Murine Model For Infrarenal Abdominal Aortic Aneurysm Development And Rupture
Authors: Elizondo-Benedetto Santiago , Zaghloul Mohamed , Arif Batool , Meade Rodrigo , Zayed Mohamed A
Published: July 2024

Introduction:
Abdominal Aortic Aneurysm (AAA) rupture is linked with increased mortality. Multiple rodent models attempt to capitulate AAA pathology to investigate a preventative cure. However, there are currently no murine models that reliably mimic AAA rupture. Compounds such as porcine pancreatic elastase (PPE), papain, β-aminopropionitrile (BAPN) and angiotensin II (ANG II) can independently promote AAA formation in rodents. We hypothesize that combining these agents can lead to reliable infrarenal AAA rupture model in mice.

Methods:
28 male C57BL/6 mice underwent aortic topical exposure to 50μL of either saline (sham), PPE, papain or both PPE+papain (20 & 10.3mg/mL respectively) for 20 minutes with a cotton ball to induce AAA degeneration. A group of 10 mice were also exposed to BAPN at 0.3% via drinking water (Fig 1A). After 3 days, another group of 5 mice was further exposed to ANG II subcutaneous pump. Weight and in vivo aortic size were measured longitudinally. Mice that died spontaneously were promptly assessed to evaluate for AAA rupture (Fig 1B). All remaining mice were sacrificed at week 4.

Results:
Compared to sham, mice exposed to papain or PPE+papain, but no BAPN or ANG II, demonstrated no change in weight gain, but significantly increased aortic diameter at week 4 (p<0.01) while administration of 0.3% BAPN demonstrated further increase in AAA diameter in both groups (p=0.002 and p<0.001, respectively; Fig 1C). All mice that received PPE+papain+0.3% BAPN as well as ANG II pump developed infrarenal AAAs and 80% developed a free rupture by day 9 (Fig 1D).

Conclusion:
We present a novel murine model that provides a high rate of infrarenal AAA development and rupture. The model not only combines matrix degradation, but also impulse risk factors known to enhance AAA rupture in humans. The feasibility and reliability of the model facilitates its use in future murine genetic and pharmacological studies that aim to understand AAA pathophysiology and methods of prevention.
Abstract 1027: Mechanical Stress Is A Non-canonical Regulator Of Inflammatory Metabolism In Immune Cells Via Netrin-1
Authors: Cyr Yannick , Newman Alexandra , Delbare Sofie Y , Wang Kristin M M , Bozal Fazli , Zhang Tracy , Schlegel Martin , van Solingen Coen , Moore Kathryn J
Published: July 2024

Atherosclerotic cardiovascular disease (CVD) is characterized by chronic inflammation of the artery wall triggered by lipoprotein deposition. Maladaptive immune responses, primarily mediated by macrophages (Mø), promote disease progression including extracellular matrix (ECM) remodeling. Emerging evidence suggests a relationship between cellular mechanics and metabolic programming, yet the role of ECM-induced mechanical stress on Mø immunometabolism remains unexplored. We hypothesized that mechanical stress induces cytoskeletal-mediated rewiring of cellular metabolism that fuels inflammation in plaque macrophages. To investigate the role of matrix-induced mechanical stress on Mø, we used an
in vitro
system that reproduces the elasticity parameters of the atherosclerotic plaque environment. Compared to stiff matrices, Mø cultured on soft matrices showed marked morphological changes, accompanied by increased inflammation and decreased mitochondrial function as assessed by RNA sequencing and flow cytometry analyses. Notably, cytoskeletal changes were associated with profound mitochondrial network reorganization and increased ROS generation. RNA-seq and ligand-receptor pairing analysis identified the neuroimmune guidance molecule netrin-1 (
Ntn1
) and its receptor
Unc5b
as most upregulated in reponse to mechanical stress. Loss-of-function studies showed that netrin-1-deficiency protects from mechanical stress-induced metabolic changes observed in WT Mø, as measured by mitochondrial network remodelling and higher mitochondrial respiration. Importantly, incubation with recombinant netrin-1 reversed the effects of genetic loss of
Ntn1
, but not
Unc5b
, suggesting receptor dependency. Mechanistically, we showed that netrin-1 controls cytoskeletal reorganization via the Rho-associated kinase (ROCK) pathway to alter mitochondrial function, and that ROCK inhibition recapitulated the effect of
Ntn1
silencing on mitochondrial potential. Collectively, our data link Ntn1-Uncb mediated changes in cytoskeletal rearrangement to alterations in mitochondrial metabolism that dictate Mø inflammation, and implicate the ECM as a non-canonical regulator of immunometabolism that perpetuates inflammation in CVD.
Abstract 1028: Cytotoxic CD4+ T Cells: Unraveling The Residual Risk In Lipid Lowering For Type 2 Diabetes Patients
Authors: Das Dayasagar , Eberhardt Natalia , Amadori Letizia , Gildea Michael , Kaur Ravneet , sajja swathy , Fernandez Dawn , Shamailova Roza , Rockman Caron B , Maldonado Thomas , Narula Navneet , Fisher Edward , Goldberg Ira J , Berger Jeffrey , Giannarelli Chiara
Published: July 2024

Type 2 Diabetes (T2D) significantly increases the risk of cardiovascular diseases (CVDs), marked by severe plaque pathology resistant to standard cholesterol-lowering treatments. While the roles of the innate immune system and macrophages in CVDs are well-established, T cell alterations, particularly involving cytotoxic CD4 T cells (CD4 CTLs), remain unexplored in human atherosclerotic plaques. In our study, we aimed to identify the phenotype and function of CD4 CTLs in blood & plaque tissue, in the context of T2D. Using unbiased single-cell RNA sequencing (scRNA-seq) analysis, we examined over 70,000 immune cells from 22 plaques of patients undergoing carotid endarterectomy (T2D: n=12, Non-Diabetic (ND): n=10), & peripheral blood mononuclear cells (PBMCs) from the CHORD study (NCT04369664) (T2D: n=12, ND: n=4). scRNA-seq analysis revealed a diverse population of CD4 T cells, like naïve, central memory (CM), effector memory (EM), tissue resident memory (Trm), T regulatory (Tregs), & two clusters of cytotoxic (CTL) T cells expressing canonical cytotoxicity-related genes. Cluster 5 expressed
GZMB, GNLY, PRF1
, whereas Cluster 10 expressed
GZMA, GZMK
. Cell-cell communication analysis demonstrated that both CTL clusters received signaling from dendritic cells and macrophages, including TREM2+ macrophages. CTL T cells reciprocally communicated with clusters of EM and EMRA CD8 T cells, underscoring their central role in plaque cell-cell communication. Notably, Cluster 5, one of these two CTL clusters, was expanded in T2D plaque samples (p=0.032) and expressed genes involved in chemotaxis, IL-2 signaling, & transcription factors (e.g.,
RUNX3, TBX21
) involved in T cell activation. The equivalent of CD4 CTL phenotype was found to be expanded in T2D individuals compared to controls, & these elevated levels persisted in T2D patients following intensive lipid-lowering therapy, indicating resistance to the treatment regimen. In summary, our study emphasizes the crucial role of CD4 CTLs in the immune dynamics of atherosclerotic plaques. The observed resistance of CTLs to lipid-lowering interventions underscores the necessity for targeted therapeutic strategies to address the distinctive immune changes in T2D-associated cardiovascular diseases.
Abstract 1030: Zeb2 Regulates Senescence And Cytotoxic Phenotype In Atherosclerotic Plaque CD8 T Cells
Authors: Eberhardt Natalia , Kaur Ravneet , Das Dayasagar , Amadori Letizia , Sajja Swathy , Bresciani Jordan , Gildea Michael , Fernandez Dawn , Rockman Caron B , Maldonado Thomas , Narula Navneet , Fisher Edward A , Goldberg Ira J , Berger Jeffrey S , Giannarelli Chiara
Published: July 2024

Circulating senescent CD8
+
CD57
+
T cells have been implicated in reduced post-myocardial infarction survival and impaired glucose homeostasis in type 2 diabetes (T2D) patients. Despite these associations, their specific role in human atherosclerotic plaques, particularly in the context of aggravated plaque pathology in T2D remains unknown. In this study, we analyzed single-cell transcriptomic data from 75,049 immune cells obtained from plaques from 12 T2D and 10 no diabetes patients enrolled in the ATHERO-IN study. We identified 14 clusters of CD8
+
T cells clusters, revealing distinct subsets such cytotoxic CD8
+
T cells with a terminally differentiated effector memory phenotype, cytotoxic/exhausted, naïve/CM and regulatory CD8
+
T cells. These clusters expressed varying degrees of senescent genes, with a notable heightened senescent transcriptional signature in T2D patients. Fibrocalcific plaques, which were prevalent in T2D patients, showed significantly enrichment in cytotoxic, inflammatory, naïve/CM, and interferon response CD8 T cell clusters. In contrast, fibroatheroma samples were predominantly enriched in MAIT-like CD8 T cells and cytotoxic/exhausted clusters. Aged CD8 T cells expanded in T2D patients exhibited elevated senescence and cytotoxic score, manifested a senescence-associated secretory phenotype expressing cytotoxic, pro-inflammatory and proatherogenic genes. Trajectory analysis and regulatory network analysis, highlighted ZEB2— a GWAS gene associated to coronary artery disease and myocardial infarction— as a potential transcription factor driving the senescence phenotype. CRISPR-based studies corroborated this, demonstrating that
ZEB2
KO CD8 T cells reduced senescence, cytotoxicity, and activation markers, underscoring the pivotal role of ZEB2 in driving the terminal differentiation and aging phenotype. In conclusion, our findings shed light on the intricate interplay between senescent CD8+ T cells and atherosclerotic plaque pathology in the context of T2D. The identification of ZEB2 as a key regulator of T cell senescence offers a potential target for therapeutic interventions aimed at mitigating the senescence-associated effects on plaque pathology and cardiovascular outcomes in T2D patients.
Abstract 1039: Endothelial Deletion Of Nck1 Reduces Atherosclerosis In Apoe-/- Mice Through The Nck1-atf3 Axis
Authors: Ben Dhaou Cyrine , Orr Wayne
Published: July 2024

The Nck family of signaling adaptors, including both Nck1 and Nck2, classically regulate cell motility during vascular development and postnatal angiogenesis. While Nck1 and Nck2 play redundant roles in this regard with deletion of both isoforms required to affect tissue remodeling, we demonstrated that global Nck1 deletion was sufficient to reduce atherosclerotic plaque formation and selective Nck1 inhibition reduces proinflammatory endothelial activation in response to oscillatory shear stress. Therefore, we sought to determine the role of endothelial Nck1 in atherosclerosis and ischemic angiogenesis using novel endothelial-specific Nck1 knockout mice. Endothelial-specific Nck1 deletion significantly reduced Western diet-induced atherosclerotic plaque formation at multiple sites compared to mice expressing endothelial-specific Cre alone or to endothelial-specific Nck2 knockout mice. This reduced plaque formation was characterized by diminished macrophage and smooth muscle incorporation despite enhanced weight gain in endothelial Nck1 knockout mice. RNA Sequencing analysis of endothelial cells in culture showed that Nck1 depletion reduces the expression of a variety of proinflammatory pathways but enhances the expression of genes involved in central carbon metabolism and proliferation. While Nck1 deletion is sufficient to blunt atherogenic inflammation, Nck1 inhibition did not affect angiogenesis or limb perfusion in the femoral artery ligation model of hind limb ischemia. To further characterized Nck1’s proinflammatory role, we utilized affinity precipitation-mass spectrometry and identified Activating Transcription Factor 3 ATF3 as a Nck1-selective binding partner. While no studies to date have examined endothelial ATF3 signaling in atherosclerosis, our preliminary data show that Nck1 is required for ATF3 expression in response to disturbed flow
in vitro
and
in vivo
, but the mechanism by which disturbed flow stimulates ATF3 expression remains unknown. ATF3 knockdown reduces disturbed flow-induced proinflammatory gene expression, suggesting that the Nck1-ATF3 signaling axis is a novel regulator of flow-induced inflammation.
Abstract 1042: Vascular Adamts7 Promotes Smooth Muscle Foam Cell Formation During Atherosclerosis
Authors: Chung Allen , Chang Hyun-Kyung , Pan Huize , Reilly Muredach P , Bauer Robert C
Published: July 2024

Human genetic studies have linked the gene ADAMTS7 with coronary artery disease. In vivo studies in mice demonstrate that ADAMTS7 is proatherogenic, active in smooth muscle cells (SMCs), and induced in response to vascular injury. However, the mechanisms governing its proatherogenicity remain unclear. We generated an SMC specific Adamts7 transgenic mouse (TG_SMC) to mimic the induction of vascular ADAMTS7. This mouse was crossed onto the LDL KO background, and after 16 weeks of western diet feeding, TG_SMC mice had a 3.5-fold increase in en-face plaque (p < 0.01) compared to controls. TG_SMC aortas showed a 3-fold increase in foam cells (p = 0.013) as measured by flow cytometry, and 80% of these cells were SMCs. Subsequent ex vivo studies show increased oxLDL uptake in TG_SMC primary SMCs (1.15x, p < 0.01).To determine the underlying mechanism, we performed RNA-seq on primary SMCs and found that TG_SMCs have increased expression of lipid handling genes (Cd36, Fabp5, Trem2). Upstream regulator analysis identified the transcription factor PU.1 as a potential mediator of these changes, and siRNA knockdown of PU.1 attenuated the gene expression increases. Additionally, siRNA knockdown of Cd36, the most upregulated gene, and a known oxLDL receptor, ameliorated the Adamts7-mediated rise in SMC lipid uptake.We next generated an Adamts7 SMC-specific knockout model to test if SMC ADAMTS7 is solely responsible for its proatherogenic effect. Surprisingly, SMC-specific Adamts7 knockout did not affect atherosclerosis in mice. We performed RNAscope on atherosclerotic LDLR KO mice and found that Adamts7 is also expressed in endothelial cells (ECs) during atherogenesis. We bred our hyperlipidemic TG model to the EC-specific Cdh5-CreERT2 and found that EC overexpression of Adamts7 also increases en face plaque burden and foam cell formation (1.5x, p<0.05), demonstrating that vascular Adamts7, regardless of the source, increases atherosclerosis. In summary, our results indicate that Adamts7 promotes lipid uptake in vascular SMCs, thus revealing a novel mechanism through which ADAMTS7 mediates atherogenesis. Our findings support the growing body of literature demonstrating that therapeutic targeting of ADAMTS7 is a potential approach to reducing atherosclerosis.
Abstract 105: Neutrophil Cracr2a Is Crucial For Calcium Signaling And Enhances Neutrophil Adhesive Function Under Thromboinflammatory Conditions
Authors: Lee Jingu , Balzraine Brett , Kuzmanova Vasilisa , Gwack Yousang , Mosher Deane , Cho Jaehyung
Published: July 2024

Ca
2+
release-activated channel regulator 2A (CRACR2A) is a Ca
2+
-binding protein that facilitates store-operated Ca
2+
entry during T cell activation. However, it is unknown whether neutrophil CRACR2A regulates neutrophil functions in inflammation. Our confocal intravital microscopy (IVM) revealed that compared to wild-type (WT) mice, myeloid-specific cracr2a conditional knockout (CKO,
cracr2a
fl/fl;Lyz-cre
) mice exhibited a significant increase in neutrophil rolling and a decrease in neutrophil adhesion, crawling, and transmigration on ear and cremaster venules in TNF-α-induced inflammation. Deletion of neutrophil cracr2a impaired β2 integrin function without affecting the expression of L-selectin, PECAM-1, and JAM-A. Neutrophil cracr2a increased Ca
2+
mobilization after stimulation with fMLP, A23187, and thapsigargin by its rapid interaction with STIM1. Due to the lack of CRACR2A inhibitors, we generated CRACR2A-deleted HL60 cells using CRISPR/Cas9. CRACR2A deletion in differentiated HL60 cells recapitulated the defects seen in cracr2a-null neutrophils, including reduced β2 integrin function and Ca
2+
mobilization after agonist stimulation. Since excessively recruited neutrophils cause brain damage in ischemic stroke, we examined whether neutrophil cracr2a contributes to stroke pathology. Using a transient middle cerebral artery occlusion (tMCAO)-induced ischemic stroke model, we found that compared to WT mice, cracr2a CKO mice exhibited improved neurological deficits and reduced infarct volume. Longitudinal 4D IVM of mouse brain revealed that neutrophil cracr2a is crucial for neutrophil adhesion and transmigration on cerebral vessels 16-24 hours after tMCAO and upregulated αMβ2 integrin on adherent neutrophils. Further, single-cell behavior analysis of 4D IVM showed that a proinflammatory subset of neutrophils arise in WT mice, but not cracr2a CKO mice, after tMCAO. We observed that the number of circulating pro-inflammatory neutrophils was markedly decreased 24 hours after tMCAO in cracr2a CKO mice compared with WT mice. Our findings indicate that neutrophil CRACR2A is a key regulator for Ca
2+
signaling and enhances neutrophil pro-inflammatory and adhesive activities in vascular inflammation and ischemic stroke.
Abstract 1050: Discrete Population Of Rare Smc Progenitor Gives Rise Exclusively To The Fibrous Cap
Authors: Gu Wenduo , Jackson William , Zhu Ashley , Li Daniel , Zhao Quanyi , Nguyen Trieu , Worssam Matthew , Ramste Markus O , Weldy Chad S , Palmisano Brian , Monteiro Joao P , Kumar Maya , Quertermous Thomas , Cheng Paul
Published: July 2024

The fibrous cap of atherosclerotic plaques is essential for the plaque stability. Rupture of the fibrous cap leads to heart attacks and strokes, and causes tens of millions of deaths globally every year. Identifying and understanding the cellular origins and plasticity of the fibrous cap is critical to developing therapeutic strategies to stabilize the atherosclerotic plaques. Utilizing the lineage tracing mouse model, we demonstrated that fibrous cap cells arise from a predefined population of cells that expresses
Notch3
at baseline. After pulse-labeling the Notch3
CreERT2
; ROSA
lsltdTomato
; Apoe
-/-
mice with tamoxifen before high fat diet and then feeding them with high fat diet for 16 weeks, Notch3 lineage traced cells stain positive for SMC markers (TAGLN, CNN1) and are nearly exclusively found at the fibrous cap in multiple atherosclerotic prone beds. Furthermore, Notch3-lineage traced SMCs and chondrogenic SMCs are mutually exclusive in the plaque, as demonstrated by the minimal overlap of tdTomato with chondrogenic SMC markers, including
Col2a1
and Sox9. Not all fibrous caps are marked, which could reflect poor Cre efficiency or new cap cells that form later. Increase tamoxifen treatment frequencing throughout high fat diet (every 10 days) significantly improved labeling efficiency and led to fully labeled lesion cap of the whole aortic root plaque. Consistently, Notch3
+
cells are committed to fibrous cap formation and excluded from the calcified portions of the lesion and acellular core. Collectively, these lineage tracing studies highlight previously unrecognized medial SMC heterogeneity in healthy vessels. Unique Notch3
+
populations of SMCs in normal media are fated to form the lesion cap. Once the Notch3 program is turned on, cells are locked into a fibrous cap fate and do not give rise to osteochondrogenic SMCs. Importantly, Notch3
CreERT2
mice can be used as a cap-specific genetic manipulation tool.
Abstract 1051: Endothelial CEPT1 Impacts Aortic Atherosclerosis And Hepatic Lipoprotein Metabolism
Authors: Hafezi Shahab , Khan Tariq J , Belaygorod Larisa , Arif Batool , Semenkovich Clay F , Zayed Mohamed A
Published: July 2024

Background:
Circulating lipids that contribute to the development of atherosclerosis are derived from very low-density lipoprotein (VLDL), produced in liver by a process requiring microsomal triglyceride transfer protein (MTTP). Since choline ethanolamine phosphotransferase 1 (CEPT1) generates phospholipids critical for membrane composition and signaling, we hypothesized that
Cept1
deficiency reduces hepatic MTTP and aortic atherosclerosis.

Methods:
Conditional endothelial cell (EC)-specific knockdown of
Cept1
(
Cept1
fl/fl
Cre+
) was performed in
ApoE
-/-
mice maintained on a 42% high-fat diet for 12 weeks, serum lipids were measured, and aorta, aortic roots, and liver tissue were harvested (Figure, A). In addition, EC-hepatocyte (HepG2 cells) signaling was evaluated using an
in vitro
co-culture assay, and
Mttp
gene expression was assessed in HepG2 cells, cultured with and without ECs treated with
Cept1
siRNA (Figure, B and C).

Results:
Compared to
ApoE
-/-
mice,
Cept1
fl/fl
Cre+
ApoE
-/-
mice had significantly reduced serum cholesterol (P < .05; Figure, D) and triglyceride (P < .01; Figure, E), and reduced atherosclerotic lesions of the aorta and aortic root (P < .01 and P < .05, respectively; Figure, F-I).
Cept1
fl/fl
Cre+
ApoE
-/-
mice also demonstrated more severe hepatic steatosis and numerically greater content of liver lipid droplets (Figure, J and K).
Cept1
knockdown in ECs co-cultured with HepG2 cells resulted in a significant decrease in
Mttp
expression (P < .01; Figure, L).

Conclusion:
Our findings suggest that deficiency of endothelial
Cept1
significantly reduces aortic atheroprogression. This may, in part, be due to a reduction in serum lipid levels via inhibition of hepatocyte MTTP driven by signals from endothelial cells. These findings open a new avenue of exploration regarding the influence of EC signaling on hepatic lipid metabolism and its impact on hyperlipidemia and atherosclerotic cardiovascular diseases.