bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhancing the Psychological Well-Being of Higher Education English Lecturers: A Qualitative Study
Authors: Ghafar Zanyar Nathir
Published: July 2024

This study explores the influence of mental health on the instructional approaches employed by higher education English lecturers in the Kurdish region. The primary objective of this study is to find effective techniques that may be implemented to enhance the psychological well-being of these teachers. The qualitative method was used by conducting semi-structured interviews with 5 lecturers at the National Institute of Technology in Kurdistan. The selection of these individuals was crucial in order to get critical insights and perspectives. The results indicate that there is a positive relationship between more significant levels of well-being and instructional practices, resulting in enhanced levels of participation, inspiration, and pleasure. The fundamental qualities that are emphasized are autonomy in teaching, competency, and effectiveness, as well as healthy student interactions. The study findings indicate that using several tactics, such as autonomy assistance, building skills, interaction, acknowledgment and feedback, and a healthy balance between work and life, is a means to enhance the overall well-being of higher education English lecturers. This study is based on a literature review on the topic of teacher well-being, offering valuable insights that may inform pedagogical administrators, staff members, and instructional programs in their efforts to cultivate a conducive atmosphere for professional development and the delivery of successful English language teaching.
Combinination Essential Ginger Oil and Acupressure Relaxation Techniques to Nausea and Vomiting
Authors: Lisnawati Ketut , Wati Ni Made Nopita , Ni Luh Putu Thrisna Dewi  , A.A Sri Sanjiwani
Published: July 2024

Introduction: Nausea and vomiting are effects of chemotherapy that can decrease in the health condition of cancer patients. Objective: The aim of this study was to determine the effect of the combination of essential ginger oil therapy and acupuncture relaxation techniques on nausea and vomiting in cancer patients undergoing chemotherapy. Methods: This research used a quasi-experimental design. The population of this study was 124 adults undergoing chemotherapy. Sampling used purpose-based sampling with a sample size of 34 respondents (17 people in the treatment group and 17 people in the control group). Nausea and vomiting were measured using the Rhodes Index Nausea, Vomitting and Retching instrument. Results: The study results showed that before receiving the intervention, the intervention group's nausea and vomiting score averaged around a score of 6, in the control group the mean score was around 6. After the intervention, the nausea and vomiting score of the intervention group was 2, in the control group 10. The Wilcoxon test showed a significant value (p < 0.001). Conclusion: The combination of essential ginger oil and Acupressure relaxation techniques helped to reduce nausea and vomiting due to chemotherapy in cancer patients. This means that an intervention given regularly can educe nausea and vomiting. Oncology nurses could give recommendation to the hospital to provide this intervention to the chemotherapy patients.
Stunting Knowledge and Attitudes among Adolescents in Banjar Pande, Sayan Village, Ubud, Gianyar: The Effect of Structured Education
Authors: Komalasari Ni Luh Putu Ratih , Oktaviani Ni Putu Wiwik , Trisnadewi Ni Wayan , Adiputra I Made Sudarma , Wahyudi Hendro
Published: July 2024

Introduction: Stunting is a chronic nutritional problem that remains a major threat to the quality of human resources. One of the efforts to break the stunting cycle is through the active role of youth. However, many adolescents are still unaware of stunting, so it is necessary to increase adolescent knowledge by providing structured education. The aim of this study was to determine the effect of structured education on adolescents' knowledge and attitudes about stunting. Methods: This study is pre-experimental research with a pretest-posttest one-group design. The sample size was 34 people, selected using a purposive sampling technique. Structured education was provided through modules and video media three times, conducted once a week for 45 minutes. Results: The results of the study showed that the average pretest score for adolescents' knowledge was 11.97, and the posttest score was 16.26. The average pretest score for adolescents' attitudes was 37.68, and the posttest score was 55.21. The results of the Wilcoxon signed-ranks test for knowledge showed a p-value of 0.000 < 0.05, and for attitudes, it also showed a p-value of 0.000 < 0.05, indicating that there was an effect of structured education on adolescents' knowledge and attitudes about stunting. Conclusion: Structured education using modules and video media is an effective method to convey messages in an understandable and engaging way, enabling the target audience to learn the material well, increasing adolescents' knowledge about stunting and encouraging the formation of positive attitudes.
The Correlation between Rooming-in and the Level of Independence of Postpartum Mothers in Caring for Newborns in the Inpatient Room of RSU Bali Royal
Authors: Agustini IGA Ratih , Prihandini Claudia Wuri , Siwi Ni Luh
Published: July 2024

Introduction: Rooming-in, or co-hospitalisation, is designed to teach postpartum mothers how to independently care for their newborns, recognise their babies' responses, and establish a strong bond. This approach is particularly crucial for mothers who exclusively breastfeed. This study aimed to determine the correlation between rooming-in and the level of independence of postpartum mothers in caring for their newborns at RSU Bali Royal, emphasising the importance of health education regarding postnatal care.
Methods: This quantitative study employed an observational, analytical, and descriptive design with a cross-sectional approach. The sample comprised all postpartum mothers who gave birth at Bali Royal General Hospital and met the inclusion criteria. A total of 92 respondents were included. Data were collected using a detailed questionnaire and analysed using the Spearman Rank Test.
Results: The study found that 53.3% of the respondents required assistance in caring for their newborns. The correlation value of 0.784 indicated a strong relationship between rooming-in and the level of independence of postpartum mothers.
Conclusion: There is a strong correlation between rooming-in and the independence level of postpartum mothers in caring for newborns at RSU Bali Royal. Promoting rooming-in practices can significantly enhance the independence of postpartum mothers.
Extent of Implementation on the Use of Body Worn Camera in Police Operations: Basis for Enhanced Action Plan
Authors: Andrade Francha , Ayon Aubrey , Cebrero Jude , Pagkaliwangan Joel , Sarte Roldan , Villa Elizabeth Buena
Published: July 2024

This study aimed to evaluate the implementation on the use of body worn camera in police operations among police officers in Cebu City. It utilized descriptive comparative analysis, employing survey methods. However, it was confined to only 11 Police Stations in Cebu City. The participants consisted of 50 police officers who were selected to utilize body worn cameras, as well as 50 community people in the study area. The selection process for both groups involved purposive sample technique. The research instrument in this study was a survey questionnaire adopted from the operational guidelines and policies on the use of body worn camera during police operations of the Philippine National Police (2018). All the strands of the implementation on the use of body worn camera in police operations were rated by the respondents based on a 5-point scale. Data were analyzed using mean, and One Way Analysis of Variance. The community and law enforcement have overwhelmingly praised body-worn cameras for their major contribution to the development of good ties between the two groups. Cameras enhance accountability and transparency, thereby boosting public trust in the police force. Nevertheless, questions regarding the implications for privacy and safety persist. The presence of cameras has a beneficial effect on both workers and the community, strengthening public trust, enhancing accountability, and decreasing the use of force. In addition, they furnish substantiation to bolster law enforcement endeavors and settle conflicts. Effective community policing depends on establishing trust, which body-worn cameras help to do.
The Effect of Influencer Marketing on Consumer Engagement and Brand Loy-alty
Authors: Yong Sean Calvin Shin Ching , Gao Xing , Poh Wen Sern
Published: July 2024

Companies spend their social media advertising on several platforms, including Facebook, Instagram, TikTok, and Xiaohongshu. The proliferation of technology and the increasing accessibility for individuals to establish careers on social media platforms have led to the emergence of influencer marketing as a strategy that can foster consumer engagement within these platforms and perhaps impact brand loyalty. This study examines the effect of influencers marketing approach in the Malaysian context, taking into consideration the growing prominence of this phenomenon. This study utilized descriptive analysis to offer generalizable recommendations for companies regarding the effect of adopting the influencer marketing approach as a strategic method. The findings of this study provide valuable insights for companies seeking to develop their marketing strategies and promote consumer engagement to expand their market position.
Grammar Express Unleashed: A Microlearning Hub Approach to Least Learned English 7 Competencies
Authors: Cunanan Maica Angelou G.
Published: July 2024

The study employed a mixed-method developmental approach to design a learning hub imbued with microlearning to address the least learned competencies in English 7. The least learned competencies were identified by considering existing test results in the locale of the researcher, which were supported by the findings of existing studies in the literature through a systematic review. The researcher assumed the roles of content creator and curator and identified relevant microlearning materials available for access. Experts validated the quality of the identified learning content through an adopted questionnaire. Afterwards, the researcher designed the Grammar Express. ICT experts were asked to validate the platform using an adapted rubric to determine whether it is suitable as a learning website. An adapted questionnaire was used to measure the level of acceptance of Grammar Express by language teachers. Meanwhile, an adapted questionnaire was used to evaluate the level of usability of the platform as perceived by the students. Based on the findings, the respondents believe that Grammar Express is useful since it offers tangible benefits and can enhance instruction. Similarly, in terms of perceived ease of use, Grammar Express requires less effort to learn and utilize. In addition, there is a strong positive attitude to incorporate Grammar Express in language teaching. Furthermore, there is a strong intention to use Grammar Express, which indicates a high level of acceptance and enthusiasm for the platform. In terms of usability, there is a strong inclination toward frequent use and displayed confidence in navigating the platform independently.
Behind the Privative Space Obstruction. Investigating Visual Perception Privacy, Downtown Algiers’ Mass Housing
Authors: Benlakehal Nadia , Lebdiri Malika , Djedi Hadjer
Published: July 2024

The present paper brings forth visual perception privacy issue in mass housing in Algiers. Considering a correlation between obstruction rates and visual perception privacy, the case study was selected to measure dwellers’ satisfaction according to Altman’s theoretical framework. A qualitative method was adopted and a semi-guided interview was conducted with 28 downtown mass housing residents in Algiers. On the other hand, to understand outsiders’ perspective regarding downtown dwellings’ perception, the qualitative method was supplemented by a survey involving 94 participants. Both methodological tools were structured according to previous non-participatory observation. Finally, the investigated area appears as being double-scaled: on an urban scale visual perception privacy measurement, people are satisfied, while on the residential scale, dissatisfaction prevails. From outsiders’ perspective, dynamic motions combined to street furniture create a visual protective shield between dwellers and unfamiliar outsiders, as the later perceive environment on a cognitive dimension, while familiar outsiders perceive it on an affective-emotional symbolic dimension.
Policy Design for Guidance and Development of Micro, Small and Medium Industries in Lubuklinggau City
Authors: Khalid Haikal Hosali Muhammad , Yogopriyatno Jatmiko , Aziz Zulhakim Abdul
Published: July 2024

This paper aims to describe the challenges and proposed strategies and policies for the Development of Micro, Small and Medium Industries in Lubuklinggau City. This research design is descriptive qualitative, data collection is done through four steps namely; observation, semi-structured interviews, documentation and conducting FGDs. Data analysis consists of two main sub-processes: (1) data reduction and pattern identification, and (2) conclusion drawing and conclusion presentation. The results of the study concluded that there are challenges faced in the activities of Guidance and Development of Micro, Small and Medium Industries in Lubuklinggau City to develop five leading sectors namely; wholesale and retail trade sector, transportation and trade sector, accommodation and eating and drinking sector, real estate sector, corporate services sector, and health services sector. Pentahelix Model Design was designed by the Lubuklinggau City Government to organize the Guidance and Development of Micro, Small and Medium Industries by involving five elements namely academics, business, community, government, and media.
Assessment of environmental degradation on the Mostaganem Plateau Algeria: A critical analysis of soil and water resources
Authors: Ouabel Habib , Taleb Mohamed Lamine , Hamadi Amina
Published: July 2024

This study underscores the importance of accounting for spatial dimensions and temperatures in evaluating payments and environmental interactions. Exploiting natural spaces can foster sustainable development, especially in vulnerable areas, but urbanization poses significant challenges. The Mostaganem Plateau, spanning 78,100 hectares with diverse landscapes, is highly sensitive to environmental pressures like abrasion. Urbanization, agriculture, industry, deforestation, and heavy port activities threaten its biodiversity. Using the plateau as a diagnostic tool helps identify the main causes of its deterioration. Analyzing agricultural and industrial impacts on soil and water highlights the severity of the situation. These analyses, along with in-depth geographic studies, reveal the complex decomposition processes driven by natural and human factors. Implementing effective political policies and management practices is crucial to counter negative influences and promote sustainable development. Protecting the Mostaganem Plateau's fragile ecosystem requires vigilant efforts from the scientific community and social and economic stakeholders. Addressing environmental and ecological impacts is essential for long-term development. Practical responses are needed to mitigate severe degradation and preserve the plateau's environmental integrity for future generations. The study emphasizes the urgent need for monitoring and intervention to safeguard this vulnerable region, ensuring its sustainability and resilience against ongoing threats. Protecting the Mostaganem Plateau's fragile ecosystem requires vigilant efforts from the scientific community and social and economic stakeholders. Addressing environmental and ecological impacts is crucial for long-term development. Practical responses are needed to mitigate severe degradation and preserve the plateau's environmental integrity for future generations.
Pemanfaatan Data Satelit Himawari-8 Dalam Analisa Kejadian Hujan Lebat Di Jombang Tanggal 1-2 Februari 2021
Authors: Sulistiyono Wahyu , Tuna Miranda Sahfira , Ramadhan Shandy Alhaq
Published: July 2024

Terjadinya peristiwa banjir di daerah Jombang pada tanggal 2 Februari 2021 menyebabkan rusaknya 38 rumah serta fasilitas umum. Peristiwa banjir disebabkan hujan dengan intensitas tinggi yang dihasilkan awan cumulonimbus. Analisa terhadap perkembangan awan konvektif diperlukan untuk melihat tahap mikrofisis awan penghasil hujan. Pada penelitian ini analisa awan konvektif serta kondisi amosfer dilakukan dengan menggunakan Satelit Himawari-8. Pengolahan dilakukan dengan metode RGB Day Convective Storm, 24H-Microphysics, Airmass serta Cloud Convective Overlay. Berdasarkan hasil penelitian, diketahui bahwa fase mikrofisis awan konvektif terjadi dalam 2 fase dengan waktu yang berbeda pada tanggal 1 dan 2. Pada analisa suhu puncak awan, selama fase perkembangan awan Cb suhu puncak awan berada dalam rentang nilai -20 sampai -60 0C. Berdasarkan citra Airmass, teramati gugusan awan konvektif berasal dari perairan Laut Jawa serta Laut Selatan. Fase puncak pembentukan awan terjadi pada jam 12.00 UTC pada tanggal 1 dan 2 Februari. Pembentukan awan konvektif mengalami peningkatan ketika nilai updraft menguat dan intensitas ketebalan awan meningkat. Sementara pada fase peluruhan ditandai dengan nilai downdraft yang lebih tinggi serta penurunan sebaran awan berdasarkan citra Day Convective Storm dan 24H-Microphysics.
Abstract 1022: Microrna-615-3p Decreases Apolipoprotein B Expression In Human Liver Cells And Its Plasma Concentrations Are Inversely Correlated With Plasma Lipid Levels
Authors: Ansari Abulaish , Yadav Pradeep K , SWATI FNU , Laine Antoine , Rimbert Antoine , Islam Shahidul , Osman Iman , Hussain M M
Published: July 2024

Plasma lipids are mainly carried in apolipoprotein B (apoB) containing lipoproteins. High levels of these lipoproteins are associated with several metabolic diseases and lowering their plasma levels is associated with reduced incidence of atherosclerotic cardiovascular disease. MicroRNAs (miRs) are small non-coding RNAs that reduce protein expression of their target genes and are potential therapeutic agents. Here, we identified a novel miR-615-3p that interacts with human 3′-UTR of apoB mRNA, induces post-transcriptional mRNA degradation, and reduces cellular and secreted apoB100 in human hepatoma Huh-7 cells. Reducing cellular miR-615-3p levels by CRISPR-sgRNA increased cellular and secreted apoB100 indicating endogenous miR regulates apoB expression. Furthermore, we found that obese subjects have lower levels of miR-615-3p than healthy individuals. Moreover, there was a significant negative correlation between miR-615-3p and plasma lipids and apoB levels. These studies indicate that miR-615-3p negatively regulates apoB and its plasma levels could serve as a biomarker for obesity and possibly other metabolic diseases.
Abstract 1043: Single-cell Characterization Of Mouse Aorta During Atherosclerosis Regression
Authors: Coronel Johana , Bashore Alex , Xue Chenyi , Zhu Lucie , Li Mingyao , Reilly Muredach P
Published: July 2024

Atherosclerosis is a cardiovascular disease characterized by the accumulation of fibro-fatty plaque in the arteries. Over half the cells in a mature atherosclerotic lesion are smooth muscle cells (SMCs) or SMC-derived cells (SDCs). SMCs can undergo a process called phenotypic switching, which involves dedifferentiation to an intermediate multipotent cell state, migration, and transdifferentiation into alternative cell types (SDCs). Increased SMC phenotypic switching is associated with worse atherosclerosis progression. However, little is known of the roles of SMCs and SDCs in atherosclerosis regression. We hypothesize that SDCs may promote plaque stability during atherosclerosis regression. In this study, we used an SMC lineage tracing mouse model (Ldlr–/–; LoxP-stop-LoxP-ZsGreen1+/-; Myh11-CreERT2+/-) that is limited to male mice. At 6 weeks of age, the mice were administered a tamoxifen diet for 1 week, followed by a week of chow diet. Mice were then fed a 16-week Western diet to induce atherosclerosis. We modeled lipid-lowering induced regression by switching to chow diet and administering HDAd-hLDLR. Mice were harvested at timepoints 0, 2, 5, 10, and 20 weeks. Arterial tissues, such as the aorta, brachiocephalic artery, and aortic root were used for histological characterization of disease and for single cell RNA-sequencing (scRNA-seq). The scRNA-seq data showed no major changes in SMC and SDC cell clusters, showing that our hypothesis was likely null. We noted an overall decrease in macrophages, and an increase in both of the two fibroblast clusters, which also had transcriptional signatures of increased proliferation and reduced apoptosis and senescence. Our histology data showed thicker fibrous caps and increased collagen, as expected in a regression model. Going forward, we will assess whether one or both fibroblast clusters are necessary for the stabilization of plaques during atherosclerosis regression.
Abstract 1049: Long Noncoding RNA Regulation Of Inflammation In Atherosclerosis
Authors: Gourvest Morgane , Barcia Jose Gabriel , Zhang Tracy , Wang Jesse , Peled Daniel , Newman Alexandra , Dupuis Samantha , Giannarelli Chiara , van Solingen Coen , Moore Kathryn J
Published: July 2024

Atherosclerosis is a chronic inflammatory disease driven by cholesterol deposition in the artery wall. Resolution of atherosclerotic inflammation and plaque regression are important clinical goals. As long noncoding RNAs (lncRNAs) are key regulators of gene expression, there is an intense interest in understanding their roles in atherosclerosis and their potential as therapeutic targets. To identify lncRNAs regulated during plaque progression and regression, we performed single cell RNA-sequencing of CD45
+
immune cells isolated from aortic arch plaques of
Ldlr
–/–
mice under conditions of plaque progression (16 wks Western diet; WD) or regression (16 wks WD + 4 wks apoB ASO). We screened for lncRNAs acting in
cis
to modify the expression of proximal protein-coding genes and identified

Caslr (
C
tsc
a
nti
s
ense
l
nc
R
NA)

, a novel lncRNA transcribed from the opposite strand of Cathepsin C gene (
Ctsc
) encoding a pro-atherosclerotic lysosomal protease.
Caslr–Ctsc
pair is upregulated in macrophages of progressing plaques and in response to pro-inflammatory stimuli (
i.e
. LPS, IL-6)
in vitro
, and conversely downregulated during atherosclerosis regression and in response to anti-inflammatory stimulus (
i.e.
IL-4). LncRNA knockdown approaches combined with transcriptomic analyses and functional assays showed that
Caslr
regulates
Ctsc
expression and associated inflammatory responses, including NF-κB signaling and inflammasome-dependent IL-1β release. RNA purification strategies combined with mass-spectrometry revealed that
Caslr
interacts with spliceosome-associated proteins to coordinate splicing of
Ctsc
mRNA. Notably,
Caslr
shares functional homology with a human lncRNA,
CASLR,
proximal to the
CTSC
gene, which regulates CTSC expression and protease activity. Human
CASLR
and
CTSC
are highly expressed in the blood of hyperlipidemic patients and their expression is coordinately decreased upon aggressive cholesterol lowering by anti-PCSK9 treatment. Collectively, our data identify
Caslr
as an evolutionary conserved lncRNA acting in
cis
to drive the expression of cathepsin C with downstream consequences on inflammatory signaling in plaque macrophages, and suggest that targeting this lncRNA may be beneficial in treating atherosclerosis.
Abstract 1063: G Protein 12 Alpha Subunit Covalent Binding To Alpha-snap And Its Potential Role In Von Willebrand Factor Secretion
Authors: Rusu Luiza , Bloem Laura J , Chen Hui , Lee Hyun , Ratia Kiira M , Jayathilaka Lasanthi , Carlier Paul R , Minshall Richard D
Published: July 2024

Rationale:
α-SNAP is part of a large 20S complex composed of 3 SNARE, 4 α-SNAP, and 6 N-ethylmaleimide sensitive factor proteins that together regulate endothelial cell Weibel Palade body fusion and exocytosis of von Willebrand factor (vWF). We previously showed that Gα12 is essential for basal and thrombin-induced vWF secretion, that GST-α-SNAP pulldown of Gα12 is dependent on N-terminal domain aa 10-15 in Gα12, and that a myristoylated 6 aa α-SNAP Binding Domain Gα12 peptide (Myr-SBD6) reduces vWF secretion, thrombo-inflammation, and lethality in septic mice. Here, using direct binding assays (Surface Plasmon Resonance (SPR), AlphaLISA, and mass spectrometry (MS)), we assessed the molecular underpinnings of SBD6 and modified higher affinity peptide (SBD15) binding directly to α-SNAP.

Methods:
GST-α-SNAP was incubated alone or with SBD6, SBD15-PEG-biotin, or SBD6 with C to S mutation and then evaluated by Maldi MS. SPR was performed for both SBD6 and SBD15 to assess binding affinity (
K
D
) and kinetics (
k
a
and
k
d
). After demonstrating the covalent linkage of α-SNAP with SBD15-PEG-biotin, cysteine mapping was performed to identify the SBD binding site on α-SNAP. Free cysteines in SBD15-biotin conjugated α-SNAP were capped with N-ethylmaleimide, disulfide bonds were reduced with DTT, reduced Cys residues were capped with iodoacetamide (IAA), and tryptic peptide digests were analyzed by LC/MS/MS.

Results:
Maldi MS studies showed that SBD15-PEG-biotin exhibited covalent binding to GST-α-SNAP. Further, proteomic analysis of Cys mapped fragments identified a peptide containing C103, KADPQEAINC
103
LMR, that was modified by NEM in control samples and shifted in the presence of SBD15, indicating modification by IAA in α-SNAP - SBD15 conjugates. Further, the C103S α-SNAP mutant was unable to bind to SBD6, and C11S-SBD15-PEG-biotin was unable to bind to α-SNAP.

Conclusion:
We propose that amino-terminal Gα12 Cys11 covalently binds to α-SNAP Cys103 and that this plays a critical role in the mechanism regulating vWF secretion. The discovery of this disulfide binding mechanism between Gα12 and α-SNAP opens up new avenues for therapeutic intervention in thrombotic disorders associated with elevated plasma vWF.
Abstract 110: Pro-thrombotic BCR-ABL Tyrosine Kinase Inhibitors Ponatinib And Nilotinib, But Not The Novel Agent Asciminb, Impair Endothelial Cell Wound Healing
Authors: Travers Richard J , Stepanian Alec , Lu Qing , Martin Gregory , Jaffe Iris Z
Published: July 2024

Introduction:
Tyrosine kinase inhibitors (TKIs) targeting ABL kinase have transformed the treatment of BCR-ABL positive leukemias. First generation imatinib is well tolerated while the newer TKIs, ponatinib and nilotinib, are more effective for leukemia but both substantially increase risk of acute arterial thrombosis, thereby limiting their clinical benefit over imatinib. Asciminib is a new BCR-ABL inhibitor with an unknown safety profile. We previously showed that BCR-ABL inhibitors that increase thrombosis in humans also cause toxicity in endothelial cells (ECs).

Hypothesis:
We hypothesize that BCR-ABL TKIs that cause EC dysfunction
in vitro
delay healing at sites of vascular injury
in vivo
as a potential mechanism of increased risk of thrombosis.

Methods:
We used the
in vitro
scratch wound assay and western blot for endothelial nitric oxide synthase (eNOS) phosphorylation in human umbilical vein (HUV)ECs and a mouse carotid artery wire injury model to assess the impact of BCR-ABL TKIs on EC wound healing capacity
in vitro
and
in vivo
.

Results:
ECs were treated with the Cmax concentration in cancer patients, as were mice, as confirmed by serum LC-MS.
In vitro
, ponatinib and nilotinib significantly decreased HUVEC wound healing compared to both imatinib and asciminib (p<0.001), with no difference between imatinib and asciminib. Ponatinib significantly decreased peNOS compared to both imatinib and nilotinib (<0.05), while asciminib was not significantly different from any of the other TKIs.
In vivo
, nilotinib significantly decreased carotid EC wound healing compared to imatinib and asciminib (p< 0.05), while ponatinib was not significantly different from any other TKIs. Carotid thrombosis after wire injury was observed in 3/26 mice treated with ponatinib, 5/23 with nilotinib, 1/21 with asciminib, and 0/26 treated with imatinib, with clot-free survival studies ongoing.

Conclusions:
Ponatinib and nilotinib cause EC toxicity by distinct mechanisms
in vitro
, which has implications for mitigating their cardiovascular risk. Asciminib causes little vascular toxicity
in vitro
or
in vivo
by these assays. Carotid thrombosis developed in some TKI treated mice, suggesting this model can be used to study the link between impaired EC healing and thrombosis.
Abstract 112: Endothelial Mixed Lineage Leukemia 1 (kmt2a/MLL1) As A Central Mediator Of Immunothrombosis During SARS-CoV-2 Infection
Authors: Parchment Nathaniel , Sharma Sriganesh , Kumar Kiran , Rocco Sabrina , Moreno Oscar Y , Luke Catherine , Gallagher Katherine A , Henke Peter , Obi Andrea T
Published: July 2024

Background:
Endotheliitis in severe SARS-CoV-2 (CoV2) results in increased cellular adhesion molecules (CAMs) and procoagulant molecules (PCMs). Cell surface CAMs increase endothelial cell (EC) interactions with circulating immune cells (IC). Increased EC-IC interactions and a procoagulant EC phenotype augment immunothrombotic risk during CoV2 infection. Prophylactic anticoagulants during CoV2 infection are associated with unacceptably high bleeding risk. New targets to treat endotheliitis are required.

Hypothesis:
We hypothesize chromatin modifying enzymes (CME)s associated with inflammatory NF-kB signaling drive transcriptional changes in endothelial cells (ECs) leading to a prothrombotic and procoagulant phenotype.

Aims:
Identify potential CMEs that drive CoV-2 endotheliitis.

Methods:
Public human single cell RNA sequencing data evaluated the pulmonary endothelial epigenetic transcriptome (PMID34876692). Murine venous ECs (mVECs), EOMA cells, and human venous ECs (HUVECs) were treated with murine beta-coronavirus (MHV-A59) or CoV2 spike protein (S1), respectively. Pharmacologic and siRNA inhibition were used to accomplish MLL1 silencing.

Results:
Humans with fatal CoV2 infection demonstrated marked increase in the KMT2/MLL class of histone methyltransferases in pulmonary ECs. mVEC infection significantly increased transcription of CME
kmt2a,
CAM
esel,
and PCMs
plau
and
f3.
Significant enrichment of H3K4 methylation and MLL1 are seen on promotors of identified genes. Inhibition of MLL1 reduced CAM and PCM transcripts. Functional EC-IC adhesion testing revealed significant decreases in monocyte adhesion with MLL1 inhibition.

Conclusion:
Kmt2a/MLL1 positively regulates CoV-2 induction of CAMs and PCMs, endothelial inflammation via positive regulation of CAM gene transcription, and resultant EC-IC interactions. MLL1 may represent a novel therapeutic target to ameliorate immunothrombotic risk during CoV-2 infection.
Abstract 1123: The Functional Roles Of Cxcr4 In Lymphangiogenesis
Authors: Bkhache Meriem , Do Long , Delgado Esteban , Oza Manisha , Liu Xiaolei
Published: July 2024

Introduction:
The lymphatic vessels play important roles in maintaining tissue fluid homeostasis, intestinal lipid uptake and immune surveillance. Malformation or dysfunction of these vessels are associated with a number of diseases, including lymphedema, inflammation, fibrosis, myocardial infarction and atherosclerosis etc. Recent studies have shown that stimulation of lymphatic vessel growth could prevent lymphedema and improve many disease pathologies, highlighting targeting lymphatic growth as a novel therapeutic approach for lymphatic related disease.

Hypothesis:
Lymphatic vessels grow by active sprouting, and subsequently mature into a vascular complex including lymphatic capillaries and collecting vessels that ensure fluid transport. However, the mechanisms regulate lymphatic sprouting and patterning are largely unknown. Here we have identified one of the key regulators and signaling pathways involved in lymphangiogenesis and function during development and disease especially CXCL12(SDF-1)/Cxcr4, a novel chemokine signaling, that plays critical roles in regulating lymphatic sprouting and morphogenesis during development.

Results:
We showed that Cxcr4 is highly expressed in migrating and sprouting lymphatic endothelial cells (LECs) during early embryonic stages. Importantly, LEC specific CXCR4 deficient embryos develop severe edema, with impaired lymphatic sprouting and migration. In addition, LEC specific Cxcr4 deficient embryos show reduced cell surface VEGFR3 but increased intracellular VEGFR3 levels in lymphatic vessels.In vitro studies showed that the blockade of Cxcr4 activity affects VEGFC-mediated LEC function. SDF-1 treatment stimulated LEC sprouting, migration and tube formation, and these effects were greatly inhibited by AMD3100 treatment which is Cxcr4 antagonist.

Conclusions:
These data suggest Cxcr4 has a novel role in regulating lymphangiogenesis, and identifying the molecular mechanism that pave the way for novel therapeutic strategies in lymphatic related disorders and cardiovascular diseases.
Abstract 1130: Tenascin-C In Arteriovenous Fistula Failure: Unraveling Venous Remodeling Dynamics
Authors: Gonzalez Luis , Zhang Weichang , Ohashi Yuichi , Vazquez-Padron Roberto , Dardik Alan
Published: July 2024

Background:
AVF are the preferred vascular access for hemodialysis, with superior patency and lower infection rates than grafts or catheters. Nevertheless, AVF patency is reduced by both early thrombosis and NIH, culminating in diminished utilization due to one-year patency rates as low as 40-50%. Tenascin-C (TnC), an ECM glycoprotein with limited adult expression, is transiently present at sites of tissue injury and inflammation. TnC has been implicated in playing a role in the development of NIH.

Hypothesis:
Sustained TnC expression contributes to NIH, potentially leading to AVF failure.

Methods:
Female and male wild-type C57Bl/6 mice, aged 9-11 weeks, had sham surgery or AVF creation by puncturing the infrarenal aorta into the inferior vena cava (IVC) with a 25-gauge needle. Doppler ultrasound confirmed AVF presence. Tissue samples were collected for IF, WB, and qPCR. Human matched vein and AVF tissue were obtained from patients undergoing planned two-stage AVF creation surgeries and analyzed with IF.
In vitro
analysis of human dermal microvascular endothelial cells (HDMEC) was performed to investigate the role of TnC in thrombomodulin expression.

Results:
TnC expression significantly increased in the AVF by day 7, returning to baseline by day 21 and reaching negligible levels by day 42. The spatial and temporal regulation of TnC expression post-AVF creation correlated with ECM remodeling and inversely with thrombomodulin. Both mice and human studies demonstrated differential regulation of TnC and thrombomodulin in patent and occluded AVF. In HDMEC, TnC knockdown suppressed NF-kB expression and increased thrombomodulin expression, while exogenous TnC had the opposite effect.

Conclusion:
TnC plays a pivotal role in driving pathological inflammation, with its expression being both temporally and spatially regulated during venous remodeling post-AVF creation. Persistent TnC expression is noted in occluded mouse and human AVF, leading to decreased thrombomodulin expression and heightened thrombogenicity. Sustained TnC expression regulates the shift from physiological to pathological venous remodeling, underscoring the crucial role of the immune response in this process and highlighting potential translational therapeutic strategies.
Abstract 1149: Exploring The Influence Of Plaque Morphology On Drug Delivery With Drug-coated Balloons Via Scanning Electron Microscopy In Peripheral Artery Disease Lesions
Authors: Csore Judit , gu Jianhua , Benfor Bright , Osztrogonacz Peter , Roy Trisha
Published: July 2024

Introduction:
Drug-coated balloons with Paclitaxel aim to address restenosis in lower extremity interventions, but concerns persist about Paclitaxel's distal embolization and drug delivery efficiency.

Hypothesis:
"Nodular" calcium exhibits poorer drug transfer than "smooth" surfaces during intervention.Aims:To investigate the impact of plaque morphology on drug transfer in advanced scanning electron microscopy (SEM).

Methods:
Vessels from amputated legs were sectioned into 5 mm samples, longitudinally halved, air-dried, gold-coated, and examined via high-vacuum SEM. Two preselected samples (one “nodular” and one “smooth” surface) were scanned with 300x300 micron field of views (FOVs), covering the surface for both samples. Drug coverage area was classified per FOV into percentage categories (minimal=0-25%, moderate=26-50%, good=51-75%, 4=76-100%) (Fig1A).

Results:
SEM analysis provided 104 and 107 300x300 micron FOVs for the “nodular” and “smooth” samples, respectively. Distribution of drug coverage categories differed significantly between “nodular” and “smooth” lesions (p<.001) (Fig1B). "Nodular" sample had lower overall drug coverage than "smooth", with a median of “good” coverage in “nodular” vs. “minimal”-“moderate” coverage in “smooth” sample” (p<.001). For the “nodular” surface, 50% of FOVs had “minimal” drug coverage (vs. 11% of “smooth” surface FOVs, p<.001). “Excellent “coverage was observed in 38.31% of “smooth” sample FOVs compared to 10.58% “nodular” FOVs (p<.001). Most of the drug in "nodular" lesion concentrated on the top of the nodule, while "smooth" exhibited uniform distribution.

Conclusions:
“Nodular” calcification may impede homogenous drug transfer, while a “smooth” surface allows for more effective drug delivery. Our approach contributes to a better understanding of drug delivery in lower extremity PVIs, ultimately guiding personalized treatment and laying the groundwork for innovative device development.
Abstract 117: AIBP Deficiency Affects Inflammarafts Formation And Macrophage Reprogramming In Atherosclerosis
Authors: Li Shenglin , Choi Sooho , Navia-Pelaez Juliana M , Miller Yury I
Published: July 2024

Background:
Excessive accumulation of cholesterol in plasma membrane of vascular macrophages leads to formation of inflammarafts - large, cholesterol-rich lipid rafts hosting inflammatory receptors assemblies - and reprograms macrophages to facilitate hyperinflammatory responses. Removal of cholesterol from membrane and storage in intracellular lipid droplets reduces macrophage inflammatory status. Apolipoprotein A-I binding protein (AIBP) has been found to mediate selective cholesterol efflux and lipid raft dynamics regulation via binding to toll-like receptor-4 (TLR4) thus reducing inflammarafts in activated macrophages. However, the underlying mechanisms driving lipid raft dynamics changes of reprogrammed macrophages remain unclear. The objective of this study was to evaluate the effect of AIBP deficiency on inflammaraft formation and macrophage reprogramming, separately in macrophage foam cells and non-foamy macrophages.

Methods:
Single-cell suspensions from the aortae of 16-week high-fat diet-fed

Apoa1bp
-/-
Ldlr
-/-

and

Ldlr
-/-

mice were gated for BODIPY-high foamy and BODIPY-low non-foamy CD45
+
F4/80
+
macrophages by flow cytometry. Lipid droplets were assessed by BODIPY fluorescence and inflammarafts were characterized by increased levels of lipid rafts and TLR4 dimers. Transcriptome and chromatin accessibility were measured by RNA-seq and ATAC-seq. Plasma lipid levels and size of atherosclerotic lesions and necrotic cores at the aortic origin were measured using standard methods.

Results:
We found a larger number of foamy macrophages in
Apoa1bp

-/-

Ldlr

-/-

mice, expressing significantly higher levels of lipid rafts and TLR4 dimers than those in
Ldlr

-/-

mice. However, there were no inflammarafts differences between non-foamy macrophages in
Apoa1bp

-/-

Ldlr

-/-

and
Ldlr

-/-

mice. The inflammarafts measures were associated with plasma cholesterol levels and size of atherosclerotic lesions and necrotic cores. Inflammatory pathways were up-regulated in
Apoa1bp

-/-

Ldlr

-/-

foamy macrophages, compared to
Ldlr

-/-

mice.

Conclusions:
Our results suggest that AIBP helps reduce inflammarafts specifically in macrophage foam cells, rendering them less inflammatory as shown by transcriptomic and open chromatin analyses.
Abstract 121: Ponatinib Promotes Endothelial Cell Inflammation And Leukocyte Trafficking Via Tumor Necrosis Factor Signaling: A Potential Mechanism Of Vascular Toxicity
Authors: Stepanian Alec , Travers Richard J , Tesfu Angelina , Lu Qing , Jaffe Iris Z
Published: July 2024

Background:
ABL tyrosine kinase inhibitors (TKI) have dramatically improved chronic myeloid leukemia (CML) survival. Due to resistance to the first ABL-TKI imatinib (Ima), most patients are treated with newer agents, including ponatinib (Pon). Although Pon improves cancer remission, the survival benefit is mitigated by a 4-fold increase in arterial thrombosis. The mechanism of Pon-induced arterial thrombosis and the safety of Asciminib (Asc), a newly approved alternative, are not known. CML patients are older and have notably elevated serum TNFa levels.

Hypothesis:
Since arterial thrombotic events occur when inflamed vascular plaques rupture, we tested the hypothesis that Pon enhances endothelial cell (EC) response to TNFa, promoting EC inflammation and leukocyte trafficking.

Methods and Results:
In vitro:
Human umbilical vein ECs (HUVECs) were treated with Pon, Ima, or Asc at concentrations observed in humans. Adhesion molecules that mediate leukocyte rolling (E- and P-selectin) and adhesion (ICAM1, VCAM1), and TNF receptors (TNFR) 1 and 2 were quantified at 24 hours by qPCR, flow cytometry (FC), and immunoblot. Pon increased TNFR2, P-selectin, VCAM, and ICAM by FC, qPCR, and immunoblot while Ima and Asc had no impact. In primary human coronary artery ECs Pon, but not Asc nor Ima, similarly increased VCAM and P-selectin mRNA. Pretreatment of HUVECs with a nonspecific TNFR inhibitor prevented Pon induction of ICAM1 and P-selectin mRNA.
In vivo:
Mice were treated with ABL TKIs by oral gavage to achieve drug plasma levels consistent with those in CML patients. After three days, leukocyte trafficking was quantified by intravital microscopy (IVM) in mouse mesenteric vessels. Pon, but not Asc, increased the number of rolling and adherent leukocytes, and increased serum TNFa by ELISA. Treatment with a TNFR inhibitor blocked Pon induced leukocyte trafficking by IVM.

Conclusion:
Pon, but not Ima nor Asc, induces expression of TNF receptors and adhesion molecules in human ECs in vitro and promotes leukocyte trafficking in vivo. Since inflamed plaques lead to acute arterial thrombosis, this mechanism could contribute to the high risk of arterial thrombosis in Pon-treated cancer patients. If so, Asc may be a safer choice as it does not induce EC inflammation.
Abstract 128: Blood TMAO Predicts Abdominal Aortic Aneurysm Growth Rate And Need For Surgical Intervention
Authors: Cameron Scott J , Li Xinmin , Benson Tyler W , Conrad Kelsey , Wang Zeneng , Helsley Robert N , Schugar Rebecca , Fleifil Salma , BUFFA JENNIFER , Maegdefessel Lars , Mani Kevin , Björck Martin , Scalise Alliefair , Cameron Scott J , Shim Sharon , Wanhainen Anders , Sharew Betemariam , Lusis Aldons J , Lyden Sean , Tang Wilson , Owens Philip , Hazen Stanley L
Published: July 2024

Background:
Patients known to have abdominal aortic aneurysms (AAAs) undergo surveillance imaging to predict both the risk for acute aortic syndromes including aortic dissection and aortic rupture, and the need for surgical intervention. Aortic rupture is a feared consequence of rapidly expanding AAAs.

Aims:
A blood biomarker that predicts risk for AAA and distinguishes slow from fast AAA growth does not exist but could be a valuable tool for risk stratification. We recently reported systemic levels of the gut microbe-generated metabolite trimethylamine N-oxide (TMAO) are associated with the presence of AAA in humans, and directly contribute to AAA progression in animal models.

Methods:
We leveraged access 805 patients in two cohorts from Europe (n=237) and the United States (n=658) undergoing serial aortic imaging surveillance. Blood TMAO concentration was measured by stable isotope dilution mass spectrometry.

Results:
TMAO was positively associated with larger baseline and follow-up abdominal aortic diameter beyond traditional factors (Adjusted OR 3.87, 95% CI, 2.70-5.58, P<0.001). In addition, TMAO predicted rapid AAA growth (HR 1.81, 95%CI, 1.23-2.67, P=0.0029). When TMAO was added to a predictive model of traditional risk factors for AAA, we saw a significant improvement in the net reclassification index (NRI, 0.43, P < 0.001; C statistic, 0.77 (0.72–0.81) with TMAO vs. NRI 0.73 (0.69–0.78) without TMAO, P =0.005). Finally, including TMAO in a predictive model for assessing need for surgical intervention among AAA patients significantly improved the prognostic performance of the model (NRI, 0.44, P < 0.001; C statistic, 0.78 (0.73–0.82) with TMAO vs. 0.75 (0.70–0.79) without TMAO, P =0.004) (Fig. 1).

Conclusions:
Elevated TMAO levels predict increased risk of AAA, and identify patients with fast-growing AAA and those who may benefit from early surgical intervention to prevent acute aortic syndromes.
Abstract 147: Monocyte-derived Extracellular Delta-like 4 Contributes To Pulmonary Hypertension Progression By Inhibiting Endothelial Notch1 Activation
Authors: Wang Shumin , Yang Yutiancheng , Wang Tengge , Rhen Jordan , Beam Nathaniel , Xu Jasmine , Jia Heqing , Ture Sara , dutheil mary , Antigny Fabrice , Montani David , Morrell Craig , Pang Jinjiang
Published: July 2024

Background:
We published that reduced Notch1 cleavage in lung endothelial cells (ECs) contributes to pulmonary hypertension (PH) development. However, the underlying mechanism of Notch1 inactivation remains elusive. We recently found enhanced circulating extracellular Delta-like ligand 4 (exDll4) from monocytes in HIV patients, the population with persistent systemic inflammation. Mobilized Notch ligands have demonstrated the ability to inhibit Notch signaling in both in vitro and in vivo studies. Hence, we hypothesize that inflammation increases monocyte-derived exDll4 and diminishes Notch1 cleavage in lung ECs, subsequently promoting the progression of PH.

Methods and Results:
Tumor necrosis factor-alpha (TNFα) is a major cytokine increase in the plasma of PAH patients and mouse models. Our
in vitro
studies revealed that TNFα induced a 17.1-fold and 23.6-fold increase of Dll4 mRNA and protein in human monocytes and a 7.2-fold elevation in exDll4 secretion. In PH mouse models induced by chronic hypoxia, Dll4 protein expression in circulating monocytes increased by 4.2 folds, and plasma exDll4 levels rose by 2.6 folds. In PAH patients, serum exDll4 was elevated by 3 folds compared to non-PAH patients. Mimicking enhanced exDll4 in PH patients; we found that exDll4 injection exacerbated PH progression in hypoxia-treated mice, accompanied by inhibited EC-Notch1 cleavage, leaky EC cell-cell junctions, and increased immune cell infiltration in the lungs. Notably, specific knock-out monocyte Dll4 in mice (mDll4 KO mice) significantly attenuated PH progression, indicated by reduced right ventricular systolic pressure and right ventricular remodeling. Mechanistically, exDll4 treatment significantly inhibited Notch1 activation and Notch target gene expression in human pulmonary microvascular ECs (HPMECs) and led to cell apoptosis, cell arrest, impaired barrier function, and enhanced inflammation.

Conclusion:
Our study demonstrates the role of monocyte-derived exDll4 in PH progression by inducing EC arrest and apoptosis and impairing barrier function in lung ECs through inhibition of Notch1 signaling. These findings imply that monocyte-derived Dll4 could be a potential therapeutic target and biomarker for PH.