bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplementary Figure 8 from Nrf2 Drives Hepatocellular Carcinoma Progression through Acetyl-CoA–Mediated Metabolic and Epigenetic Regulatory Networks
Authors: Xi Caixia , Pang Junfeng , Barrett Amanda , Horuzsko Anatolij , Ande Satyanarayana , Mivechi Nahid F. , Zhu Xingguo
Published: October 2023

S8. GCK and ALDOA gene expression in glucose-deprived shNRF2 HepG2 cells after energy-refeeding.
Supplementary Figure 7 from Nrf2 Drives Hepatocellular Carcinoma Progression through Acetyl-CoA–Mediated Metabolic and Epigenetic Regulatory Networks
Authors: Xi Caixia , Pang Junfeng , Barrett Amanda , Horuzsko Anatolij , Ande Satyanarayana , Mivechi Nahid F. , Zhu Xingguo
Published: October 2023

S7. Nrf2 suppression decreases liver tumor progression.
Supplementary Figure 4 from Nrf2 Drives Hepatocellular Carcinoma Progression through Acetyl-CoA–Mediated Metabolic and Epigenetic Regulatory Networks
Authors: Xi Caixia , Pang Junfeng , Barrett Amanda , Horuzsko Anatolij , Ande Satyanarayana , Mivechi Nahid F. , Zhu Xingguo
Published: October 2023

Supplementary Figure 4. NRF2 silencing shows no effect on histone acetylation or acetyl-CoA.
Fig S4 from HCK Promotes High-Grade Serous Ovarian Cancer Tumorigenesis through CD44 and NOTCH3 Signaling
Authors: Khella Christen A. , Franciosa Lucyann , Rodirguez-Rodriguez Lorna , Rajkarnikar Resha , Mythreye Karthikeyan , Gatza Michael L.
Published: October 2023

The effect of GSIs on NOTCH3 ICD expression in HGSOC cells following HCK overexpression.
Fig S2 from HCK Promotes High-Grade Serous Ovarian Cancer Tumorigenesis through CD44 and NOTCH3 Signaling
Authors: Khella Christen A. , Franciosa Lucyann , Rodirguez-Rodriguez Lorna , Rajkarnikar Resha , Mythreye Karthikeyan , Gatza Michael L.
Published: October 2023

The source of elevated HCK signature is not due to a bias in tumor purity or immune infiltration.
Supplemental material and methods from Nrf2 Drives Hepatocellular Carcinoma Progression through Acetyl-CoA–Mediated Metabolic and Epigenetic Regulatory Networks
Authors: Xi Caixia , Pang Junfeng , Barrett Amanda , Horuzsko Anatolij , Ande Satyanarayana , Mivechi Nahid F. , Zhu Xingguo
Published: October 2023

Supplemental material and methods
Supplemental Table 1 from Nrf2 Drives Hepatocellular Carcinoma Progression through Acetyl-CoA–Mediated Metabolic and Epigenetic Regulatory Networks
Authors: Xi Caixia , Pang Junfeng , Barrett Amanda , Horuzsko Anatolij , Ande Satyanarayana , Mivechi Nahid F. , Zhu Xingguo
Published: October 2023

Supplemental Table 1. DNA Oligos
Supplemental Table 4 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

Mutant ESR1 generation: gBlock used for template amplification
Supplemental Table 3 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

Differentially expressed genes from in vivo RNA-seq experiments
Supplemental Table 1 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

Differentially expressed genes from in vitro RNA-seq experiments
Supplemental Figure 7 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

In vivo exploration of novel therapeutic targets in ER active and mutant endometrial cancer
Supplemental Figure 5 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

ER-Y537S and ER-D538G mutants drive proliferative gene expression signatures in vivo
Supplemental Figure 3 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

ER binding is altered by the Y537S and D538G mutations
Supplemental Figure 1 from Allele-Specific Gene Regulation, Phenotypes, and Therapeutic Vulnerabilities in Estrogen Receptor Alpha–Mutant Endometrial Cancer
Authors: Blanchard Zannel , Rush Craig M. , Arnesen Spencer , Vahrenkamp Jeffery M. , Rodriguez Adriana C. , Jarboe Elke A. , Brown Callie , Chang Matthew E.K. , Flory Mark R. , Mohammed Hisham , Modzelewska Katarzyna , Lum David H. , Gertz Jason
Published: October 2023

Validation and gene expression analysis of ER mutant models
Supplementary Figure 8 from Continuous Administration of Anti-VEGFA Antibody Upregulates PAI-1 Secretion from Ovarian Cancer Cells via miR-143-3p Downregulation
Authors: Yagi Taro , Sawada Kenjiro , Miyamoto Mayuko , Shimizu Aasa , Oi Yukako , Toda Aska , Nakamura Koji , Kinose Yasuto , Kodama Michiko , Hashimoto Kae , Kimura Tadashi
Published: October 2023

Supplementary Figure 8 shows the representative images of immunohistochemical staining of a human normal kidney for PAI-1 and CD31.
Supplementary Figure 6 from Continuous Administration of Anti-VEGFA Antibody Upregulates PAI-1 Secretion from Ovarian Cancer Cells via miR-143-3p Downregulation
Authors: Yagi Taro , Sawada Kenjiro , Miyamoto Mayuko , Shimizu Aasa , Oi Yukako , Toda Aska , Nakamura Koji , Kinose Yasuto , Kodama Michiko , Hashimoto Kae , Kimura Tadashi
Published: October 2023

Supplementary Figure 6 shows that the transduction of miR-143-3p does not attenuate in vitro tube formation in HUVECs incubated with the culture media of SKOV3ip1 cells.
Supplementary Figure 2 from Continuous Administration of Anti-VEGFA Antibody Upregulates PAI-1 Secretion from Ovarian Cancer Cells via miR-143-3p Downregulation
Authors: Yagi Taro , Sawada Kenjiro , Miyamoto Mayuko , Shimizu Aasa , Oi Yukako , Toda Aska , Nakamura Koji , Kinose Yasuto , Kodama Michiko , Hashimoto Kae , Kimura Tadashi
Published: October 2023

Supplementary Figure 2 shows that PAI-1 enhanced in vitro tube formation not only in the absence of anti-VEGFA antibody but also in the presence of anti-VEGFA antibody.
Supplementary Figure 4 from Simultaneous Targeting of DNA Polymerase Theta and PARP1 or RAD52 Triggers Dual Synthetic Lethality in Homologous Recombination–Deficient Leukemia Cells
Authors: Sullivan-Reed Katherine , Toma Monika M. , Drzewiecka Malgorzata , Nieborowska-Skorska Margaret , Nejati Reza , Karami Adam , Wasik Mariusz A. , Sliwinski Tomasz , Skorski Tomasz
Published: October 2023

S4. Targeting Polθ + PARP and Polθ + RAD52 induced dual synthetic lethality against HR -deficient MPN cells.
Supplementary Figure 1 from Continuous Administration of Anti-VEGFA Antibody Upregulates PAI-1 Secretion from Ovarian Cancer Cells via miR-143-3p Downregulation
Authors: Yagi Taro , Sawada Kenjiro , Miyamoto Mayuko , Shimizu Aasa , Oi Yukako , Toda Aska , Nakamura Koji , Kinose Yasuto , Kodama Michiko , Hashimoto Kae , Kimura Tadashi
Published: October 2023

Supplementary Figure 1 shows Kaplan–Meier plotter analysis of genes that were upregulated during anti-VEGFA treatment.
Supplementary Figure 5 from Hierarchical Phosphorylation of HOXB13 by mTOR Dictates Its Activity and Oncogenic Function in Prostate Cancer
Authors: Chen Yonghong , Dufour Catherine R. , Han Lingwei , Li Ting , Xia Hui , Giguère Vincent
Published: October 2023

Supplementary Figure 5. HOXB13 transcriptional activity depends on its phosphorylation status.
Supplementary Figure 4 from Hierarchical Phosphorylation of HOXB13 by mTOR Dictates Its Activity and Oncogenic Function in Prostate Cancer
Authors: Chen Yonghong , Dufour Catherine R. , Han Lingwei , Li Ting , Xia Hui , Giguère Vincent
Published: October 2023

Supplementary Figure 4. Androgen response dependency on HOXB13 phosphorylation.
Table S5 from Analysis of Several Common APOBEC-type Mutations in Bladder Tumors Suggests Links to Viral Infection
Authors: Rao Nina , Starrett Gabriel J. , Piaskowski Mary L. , Butler Kelly E. , Golubeva Yelena , Yan Wusheng , Lawrence Scott M. , Dean Michael , Garcia-Closas Montserrat , Baris Dalsu , Johnson Alison , Schwenn Molly , Malats Nuria , Real Francisco X. , Kogevinas Manolis , Rothman Nathaniel , Silverman Debra T. , Dyrskjøt Lars , Buck Christopher B. , Koutros Stella , Prokunina-Olsson Ludmila
Published: October 2023

Table S5: Associations between driver mutations in BC and common risk factors
TABLE 2 from Change in Neighborhood Socioeconomic Status and Adherence to the Cancer Prevention Lifestyle Guidelines in Hispanic/Latino Adults: Results from the HCHS/SOL Study
Authors: Pichardo Margaret S. , Pichardo Catherine M. , Talavera Gregory A. , Gallo Linda C. , Kuo Charlene C. , Castañeda Sheila F. , Chambers Earle C. , Daviglus Martha L. , Pirzada Amber , Perreira Krista M. , Sotres-Alvarez Daniela , Peña Ortiz Tania Yadhira , Plascak Jesse J.
Published: October 2023

Weighteda mean and SE for measures of change in neighborhood SES of U.S. Hispanic/Latino adults, by American Cancer Society Guideline Adherence
Factors Associated with Complications after Total Pharyngo-Laryngo-Esophagectomy and Free Jejunal Flap Reconstruction
Authors: Ikuta Kento , Suyama Yoshiko , Fukuoka Kohei , Morita Maki , Kimura Yuka , Umeda Ryunosuke , Kanayama Haruka , Ohga Makoto , Nakagaki Makoto , Fukuhara Takahiro , Fujiwara Kazunori , Yagi Shunjiro
Published: October 2023

Introduction: Free jejunal flap (FJF) reconstruction is a standard procedure for pharyngeal and cervical esophageal defects resulting from head and neck cancer resection. However, improvements in patients’ quality of life after surgery require a further statistical approach. Methods: An observational, retrospective, multivariate analysis was designed to report the incidence of postoperative complications and their association with clinical factors in 101 patients who underwent total pharyngo-laryngo-esophagectomy and FJF reconstruction for head and neck cancer at a university hospital between January 2007 and December 2020. Results: Postoperative complications were observed in 69% of patients. In the reconstructive site, anastomotic leak, observed in 8% of patients was associated with vascular anastomosis in the external jugular vein system (age-adjusted odds ratio [OR]: 9.05, p = 0.044) and anastomotic stricture, observed in 11% of patients was associated with postoperative radiotherapy (age-adjusted OR: 12.60, p = 0.02). Cervical skin flap necrosis was the most common complication (34%) and was associated with vascular anastomosis on the right cervical side (age- and sex-adjusted OR: 4.00, p = 0.005). Conclusion: Although FJF reconstruction is a useful procedure, 69% of patients suffer a postoperative complication. We suppose that anastomotic leak is related to the low blood flow resistance of the FJF and inadequate drainage of the external jugular venous system, and anastomotic stricture is related to the vulnerability of the intestinal tissue to radiation. Furthermore, we hypothesized that the location of the vascular anastomosis may affect the mesenteric location of the FJF and the dead space in the neck, leading to the development of cervical skin flap necrosis. These data contribute to increasing our knowledge about postoperative complications related to FJF reconstruction.
Prolonged Duration of Air-Dry Fixation of Cervical Smears Produces Superior Cytomorphological Staining Quality over Conventional Wet-Fixed Smears
Authors: Danquah kwabena Owusu , Adankwah Ernest , Dadzie Headtlove E. , Gyamfi Daniel , Adjei Ernest Akwasi , Ossei  Sampene Paul Poku , Morhe Emmanuel
Published: October 2023

Introduction: Immediate fixation of smears in 95% alcohol for Pap staining is commonly used for cytopathological diagnosis of cancers and other diseases. Few research studies have investigated the comparative outcomes of alcohol wet fixation and rehydration of air-dried smears, indicating that air-dried rehydrated (ARF) smears are viable alternative to wet-fixed (WF) smears. However, there are no or few investigations on the effects of long durations of air-drying fixation on cytomorphological staining quality. Materials and Methods: 124 cervical smears were obtained from the Family Planning Unit of Komfo Anokye Teaching Hospital in Kumasi, Ghana. Quadruple smears were WF and air-dried for 2 h, 4 h, and 8 h prior to rehydration with normal saline and fixation (ARF). All the smears were stained with Papanicolaou stain, examined microscopically for their cytomorphological features, and scored. Cytomorphological scores were statistically analyzed using SPSS software. Results: No significant difference in cytolysis, cell border, nuclear border, chromatin, and cellularity between WF and ARF was observed. However, significant differences in cytoplasmic staining quality (p value <0.001) and the absence of RBCs (p value <0.001) were observed in the 4-h ARF. The absence of RBCs in the ARF smears rendered a clearer background than in the wet fixation. Conclusions: ARF, Pap-stained smears showed comparably superior cytomorphological features to those of WF smears. The 8-h ARF smears produce crispy chromatin and excellent background, making them suitable for bloody cytological samples.
Survival and Swallowing Function after Primary Radiotherapy versus Transoral Robotic Surgery for Human Papillomavirus-Associated Oropharyngeal Squamous Cell Carcinoma
Authors: Hughes Ryan T. , Levine Beverly J. , May Nelson , Shenker Rachel F. , Yang Jae H. , Lanier Claire M. , Frizzell Bart A. , Greven Kathryn M. , Waltonen Joshua D.
Published: October 2023

Introduction: The aim of this study was to investigate the impact of primary transoral robotic surgery (TORS) versus radiotherapy (RT) on progression-free survival (PFS), overall survival (OS), and 1-year swallowing function for patients with early-stage HPV-associated oropharyngeal squamous cell carcinoma (OPSCC). Methods: Patients with stage I-II (AJCC 8th Ed.) HPV-associated OPSCC treated with TORS followed by risk-adapted adjuvant therapy or (chemo)radiotherapy between 2014 and 2019 were identified. PFS, OS, and swallowing outcomes including gastrostomy tube (GT) use/dependence, and Functional Oral Intake Scale (FOIS) change over 1 year were compared. Results: One hundred sixty-seven patients were analyzed: 116 treated with TORS with or without adjuvant RT and 51 treated with RT (50 chemoRT). The RT group had more advanced tumor/nodal stage, higher comorbidity, and higher rates of concurrent chemotherapy. There were no differences in 3-year PFS (88% TORS vs. 75% RT) or OS (90% vs. 81%) between groups, which persisted after adjusting for stage, age, and comorbidity. GT use/dependence rates were higher in the RT group. Mean (SD) FOIS scores in the TORS group were 6.9 (0.4) at baseline and 6.4 (1.0) at 1 year, compared with 6.7 (0.6) and 5.6 (1.7) for the RT group. Only clinical nodal stage was found to be significantly associated with FOIS change from baseline to 1 year. Conclusion: There were no differences in PFS or OS between patients treated with primary TORS or RT for early-stage HPV-associated OPSCC. Clinical N2 status is associated with FOIS change at 1 year and may be the major factor affecting long-term swallowing function, irrespective of primary treatment modality.
PERBANDINGAN KARAKTERISTIK KLINIS PASIEN COVID 19 VARIAN OMICRON DAN VARIAN DELTA DI RUMAH SAKIT UMUM DAERAH DR. MOEWARDI SURAKARTA
Authors: Permana Septian Adi , Purwoko P , Thamrin Muhammad Husni , Anggoro Pandu
Published: October 2023

PENDAHULUAN

Perkembangan SARS-CoV-2 telah ditandai dengan pengenalan "Variant Of Concern," atau perubahan sifat virus seperti penularan penyakit dan antigenisitas. VOC Delta SARS-CoV-2 40% -60% lebih mudah menular daripada VOC Alpha (B.1.1.7) dan dapat dikaitkan dengan peningkatan risiko rawat inap.

TUJUAN

Untuk mengetahui karakteristik klinis pasien COVID-19 varian Delta dan Omicron di High Care Unit (HCU) dan Intensive Care Unit (ICU) Rumah Sakit Dr Moewardi.

METODE

Penelitian deskriptif analitik kuantitatif dengan pendekatan retrospektif yang dilakukan pada Juni 2021-Maret 2022. Sampel penelitian adalah pasien terkonfirmasi positif COVID-19 yang di rawat di ICU dan HCU isolasi pada Agustus-Oktober 2021 dan Januari-Maret 2022. Diukur tanda vital, komorbid, hasil laboratorium, status intubasi dan mortalitas.

HASIL

Angka kematian pasien yang terinfeksi Covid-19 varian delta dan varian omicron masing-masing adalah 10% dan 10,8%. Varian Delta lebih signifikan berdampak terhadap CVA (p=0.001) dan penyakit jantung (p<0.001) dibandingkan pada CVA (p=0.015) dan penyakit jantung (p=0.190) pada pasien SARS-CoV-2 varian Omicron. Varian Omicron lebih signifikan berdampak terhadap sepsis (p=0.012) dibandingkan varian Delta (p=0.026).

KESIMPULAN

Tidak didaptakan perbedaan angka mortalitas antara varian COVID-19 omicron dan delta. Kematian pada kelompok omicron sebagian besar disebabkan penyakit komorbid, sedangkan pada kelompok delta, kematian disebabkan karena COVID-19 menyebabkan disfungsi endotel.
E-Learning Competencies for Elementary School Teachers in Botolan, Zambales, Philippines
Authors: Datugan Kimberly V. , Cabal Esmen M.
Published: October 2023

Teachers' communication has been forced online due to the spread of COVID-19. Although technology advancement has made many things possible, the transition to online learning isn't simple. This descriptive study measured the level and challenges encountered in E-learning among one hundred fifty elementary school teachers. This study investigated the level and challenges of E-learning among 150 elementary school teachers. The research measured competencies such as individual media competence, critical media competence, and educational design competence using a survey questionnaire. The majority of respondents were female teachers in the adult stage, with teaching experience and educational qualifications. The teachers considered themselves competent in E-learning and disagreed with the presence of challenges. However, there was a significant difference in challenges based on the highest educational attainment. An online training program was proposed to enhance E-learning competencies, and collaboration with agencies and software companies was suggested for technical assistance. Adequate technical and peer support during E-learning implementation were recommended. School heads were advised to focus on professional development emphasizing technology integration, and teachers were encouraged to utilize visualization tools. Collaboration between school heads and teachers was emphasized for the proposed training program and strategies to address challenges.
Interpersonal, Leadership, and Supervisory Skills of Public School Heads
Authors: Trongco Goldie P. , Benolirao Eingilbert C.
Published: October 2023

This study determined the level of competency of the Elementary School Heads in the District of Sibonga, along with the aspects of interpersonal, leadership, and supervisory skills during the school year 2020-2021 a basis for the management training framework. The areas of concern included the perceived level of competency in terms of interpersonal, leadership, and supervisory skills; the significant difference in the perception of the respondent groups of the three variables; the participants’ perspectives towards the three variables; and the management training framework. The result of the study showed that the respondents showed the same level of competency. Most of them have maintained a competent level of perception. Based on the result, respondents showed the same level of perception as to the different variables presented and no significant difference between them. It can be concluded that the passion and enthusiasm of our teachers and school heads are still maintained and even doubled due to the sudden change in learning approach. It is recommended that there is a need to assess the leadership style and situational favorableness of our teachers and school heads. With that, the Management Training Framework is being proposed.
Motives and Barriers to Exercise among Underweight Filipino College Students
Authors: Manasan Patrick L. , Quitor Cristine G. , Ronquillo Vincent P. , Dabu Darelle C. , Dela Cruz Jennilyn T. , Sode Ma. Angela V. , Cruz Salvador Jr. L.
Published: October 2023

In many cases, underweight students may be left behind due to poor nutrition, health issues, or lack of access to resources that would help them thrive and achieve their academic goals. This condition is often overlooked, but it can have serious health consequences. In this paper, the researchers used the free-listing method to ascertain the perceived motives and barriers to exercise among underweight Filipino university students. This quantitative-descriptive study comprised 300 underweight Filipino college students enrolled in ‘PATHFIT-2: Exercise-Based Fitness Activities' for first-year students during their Second Semester of the Academic Year 2022-2023 from the main campus of a state university in Pampanga, Philippines, who were classified as having BMIs below the normal weight. Furthermore, EMI-2 from Mullan et al. (1997) subscales were used to categorize responses in the free list for motives, whereas Myers and Roth's (1997) subscales were utilized to categorize barriers. The top five reported motives for underweight students were "positive health," "strength and endurance", "ill health avoidance", "appearance", and "weight management". While "competition", "nimbleness", "affiliation", and "health pressures" did not elicit a response.  On the other hand, most respondents do not have enough time and are too lazy to exercise since it may interfere with their school-related tasks. The findings provided educational institutions with precise recommendations on how to establish exercise programs that are in line with what motivates students and examine any barriers that may prevent such physical activity engagement.
The Traditional Games of Macabebe, Pampanga
Authors: Lacson Kylene S. , Matuguina Daisy Mae A. , Cortes Yna Frances S. , Guinto Geraldine N. , Caoleng Clarisse Faye B. , Carlos Mary Chantelle O.
Published: October 2023

The study aimed to discover the undocumented traditional games within the Municipality of Macabebe, Pampanga, the Philippines. The data findings are to be contextualized in the educational implications of the Physical Education curriculum. With the use of a qualitative research approach, particularly a focused ethnography, a total of 16 key informants with age ranges from 61 to 102 years old participated in the semi-structured interview. After the data process, there were 11 traditional games found, five in the coastal area classified as moderately vigorous intensity when playing while six games in the land were vigorous. Using the Braun and Clarke thematic analysis, three themes were explicated. The first theme encompasses the people in Macabebe’s way of life and how the participants provide basic needs to their families. The second theme describes the origin of undocumented conventional games. The third theme states the educational implications of the Physical Education curriculum among the students. Relatively, the study provides data that enumerates the undocumented traditional games in Macabebe to be integrated into the curriculum.
Abstract 102: Hyperlipidemia-induced Hematopoiesis is Repressed by MLKL in Endothelial Cells of the Splenic Niche
Authors: Rasheed Adil , Dennison Taylor , Robichaud Sabrina , Wyatt Hailey , Nguyen My-Anh , Geoffrion Michele , Marouf Yacine , Baxi Adir , Lee Richard , Kazan Hilal , van Solingen Coen , Ouimet Mireille I , Rayner Katey J
Published: October 2023

The mixed lineage kinase domain-like protein (MLKL) is well-known for its role in the execution of necroptotic cell death. We showed that antisense oligonucleotide knockdown of MLKL (
Mlkl
KD) in
Apoe
-knockout mice reduces plaque necrotic core, as predicted, but not total plaque area. Additionally,
Mlkl
KD expanded splenic hematopoietic stem and progenitor cells (HSPCs) and mature myeloid cells known to drive atherogenesis. Thus, we
hypothesized
that MLKL limits splenic hematopoiesis and thereby immune cell contributions to the growing atheroma. To test whether hematopoietic or non-hematopoietic MLKL expression was responsible for HSPC expansion, we transplanted
Ldlr
-knockout mice with
Mlkl
-knockout bone marrow and found no changes in hematopoiesis, suggesting that MLKL functions in other cells in the splenic niche. Flow cytometry and immunofluorescence staining during a time course of atherosclerosis revealed a decrease of splenic endothelial cells that was correlated with a reduction of MLKL endogenously or by
Mlkl
KD, suggesting that MLKL maintains splenic endothelial cells. Both
in vivo
and
in vitro
we observed increased lipid accumulation and concomitant defects in cell cycle in
Mlkl
KD splenic endothelial cells. Similar to our previous work in macrophages,
Mlkl
KD led to an accumulation of CHMP4B in cultured splenic endothelial cells, suggesting aberrant endocytic trafficking, which has also been linked to dysfunctional proliferation. Furthermore, there was greater production of immune cells derived from transplanted CD45.1 HSPCs in
Mlkl
KD recipient spleens. Co-culture with
Mlkl
KD splenic endothelial cells increased HSPC expression of pStat5, a marker of activation, which was coupled with an increase in myeloid colony formation, as compared to co-culture with control splenic endothelial cells. Remarkably no changes in HSPC regulation were found upon
Mlkl
KD in bone marrow endothelial cells. In conclusion, we demonstrate a novel role for MLKL in regulating the balance of HSPCs, specifically through preservation of splenic endothelial cells that repress hematopoiesis, highlighting the importance of spleen lipid metabolism and its impact on inflammation that potentiates atherogenesis.
Abstract 106: Single Cell High Dimensional Analysis of Human Peripheral Blood Mononuclear Cells Reveals Unique Intermediate Monocyte Subsets Associated with Sex Differences in Coronary Heart Disease
Authors: Komaravolu Ravi K , Chatterjee Nandini , Alimadadi Ahmad , McNamara Coleen A , Taylor Angela M , Manichaikul Ani , Miller Yuri I , Ley Klaus , Hedrick Catherine C
Published: October 2023

Monocytes are innate immune cells that are defined by three subsets: classical (cMo), intermediate (iMo) and non-classical (nMo). How these subsets become more heterogeneous in phenotype and function in CVD progression remains elusive. We employed single-cell transcriptome and protein analysis to define how human monocytes differ in subjects with moderate to severe CVD. Peripheral blood mononuclear cells (PBMC) were obtained from 61 human subjects, undergoing coronary angiography as part of the Coronary Assessment of Virginia (CAVA) Cohort. Coronary atherosclerosis severity was quantified using Gensini Score (GS). We analyzed 487 immune-related genes and 49 surface proteins at the single cell level in PBMC from each subject using the BD-Rhapsody platform. WNN Seurat 4.0 was used for high dimensional analysis. Data was normalized using GLMM to control for covariates (age, sex, diabetes, and statin use). We identified cMo, iMo, nMo, plasmacytoid DC, and classical DC. We found a significant increase only in the frequencies of intermediate (iMo) monocytes in subjects with high CVD (GS>30) compared to subjects with low CVD (GS<6) (
p
=0.004). Spearman correlation analysis with GS from each subject revealed a positive correlation with iMo frequencies (r=0.314,
p
=0.014) and further showed sex-dependent positive correlation in female patients (r=0.663,
p
=0.0037). Interestingly, cMo, thought to be potently atherogenic, did not correlate with CVD severity. Further analysis identified 3 iMo subsets, distinguished by expression of HLA-DR, CD86, CXCR3, and PD-L1. We found that frequencies of two of them, iMo_CXCR3
+
PDL1
lo
(
p
=0.0003) and iMo_HLA-DR
+
CD86
int
(
p
=0.019), were associated with CVD severity. The iMo_CXCR3
+
PDL1
lo
immunoregulatory subset was positively correlated with GS in both females (r=0.66,
p
=0.004) and males (r=0.315,
p
=0.037). However, the iMo_HLA-DR
+
CD86
int
subset was positively correlated only in females (r=0.55,
p
=0.022), illustrating substantial heterogeneity among monocyte subsets in a sex-dependent manner. In sum, we identified novel sex- and CVD-dependent iMo subsets in subjects with CVD. Future functional analysis of iMo populations should provide a deeper understanding of long-known sex-differences in CVD progression.
Abstract 107: CaMK4 is a Novel Regulator of Myeloid Phenotype and Function in Atherosclerosis
Authors: Gonzalez Azuah L , Youwakim Cristina , Dungan Matthew M , Fuller Kristin , Michell Danielle L , Cavnar Ashley , Rahman Jamshed , Vickers Kasey C , Brown Jonathan D , Doran Amanda C
Published: October 2023

Background:
Chronic inflammation is a major driver of atherosclerotic cardiovascular disease, and therapeutics that target inflammation reduce clinical cardiac events beyond levels seen with conventional strategies targeting cholesterol alone. Recent work by our group and others has demonstrated that advanced atherosclerosis is also characterized by the failure of an active repair process termed ‘inflammation resolution’. Strategies that boost resolution and break the cycle of chronic inflammation promotes plaque stability.

Hypothesis:
Our review of publicly available RNAseq data revealed an increase in expression of Calcium/calmodulin-dependent protein kinase 4 (CaMK4) in advanced/unstable regions of plaque within human carotid arteries as well as in myeloid cells derived from atherosclerotic murine aortas. Therefore, we hypothesized that CaMK4 promotes inflammation and impairs resolution.

Results and Methods:
Control and

Camk4
-/-

mice were injected with AAV-8 PCSK9 virus and fed a high-fat high-cholesterol Western diet for 12 weeks. Cross-sectional analysis of aortic roots showed that

Camk4
-/-

mice had significantly less plaque burden than Controls. Flow cytometric analysis revealed elevated monocyte numbers in

Camk4
-/-

mice compared to Control mice. Further analysis demonstrated increased skewing of

Camk4
-/-

monocyte populations toward a Ly6c
low
subset, indicating a more pro-reparative phenotype compared to Control mice. We considered whether CaMK4 may regulate a gene signature driving monocyte conversion and found that

Camk4
-/-

monocytes expressed higher levels of
Nr4a1, Cebpb,
and
Klf2,
which have been shown to promote conversion to Ly6c
low
monocytes. As Ly6c
low
monocytes give rise to pro-reparative macrophages, we generated bone marrow-derived macrophages and found that loss of CaMK4
in vitro
led to higher levels of pro-reparative cytokines being produced in response to LPS stimulation as well as more efficient clearance of apoptotic cells when compared to Controls.

Conclusions:
Overall, these findings suggest that CaMK4 regulates myeloid phenotype
in vitro
and
in vivo
and that loss of CaMK4 promotes pro-resolving macrophage function. Therefore, targeting CaMK4 may offer a unique way to target atheroprogression.
Abstract 108: Il-1β Inhibition Promotes Fibrous Cap Thickening and Accumulation of Fibroblast Like Cells in Atheromas in Clonal Hematopoiesis
Authors: Fidler Trevor P , Kim Eunyoung , Hardaway Brian , Xue Chenyi , Abramowicz Sandra , Xiao Tong , Maegdefessel Lars , Wang Nan , Tall Alan
Published: October 2023

Clonal hematopoiesis (CH), a highly prevalent condition in the elderly, arises from somatic mutations that endow a survival advantage to hematopoietic stem cells. In CANTOS IL-1β inhibition was particularly beneficial in subjects with
TET2
CH. Amongst the common genetic variants giving rise to CH,

JAK2
V617F

(

JAK2
VF

) gain of function and
TET2
loss of function mutations have been shown to increase macrophage inflammasome activation and atherosclerosis in mice. We have shown that macrophages harboring
Jak2
mutations have increased AIM2 inflammasome activation and pyroptosis which promoted features of unstable atherosclerotic lesions marked by prominent necrotic cores and an accumulation of inflammatory macrophages. Inflammasome inhibition led to a thickening of fibrous caps, indicating plaque stabilization. scRNA-Seq analysis of human carotid plaques indicates that myeloid cells were enriched for inflammasome components while endothelial, fibroblast, and smooth muscle cell (SMC) like cells were enriched for the IL-1 receptor suggesting IL-1-mediated cross-talk from myeloid cells to stromal cells. Here we investigated this cross-talk using IL-1β antibodies in mice modeling

Jak2
VF

or
Tet2
CH and expressing Myh11-Cre/ZS-green in order to fate map SMC-derived cells. ScRNA-Sequencing of green and non-green cells revealed that IL-1β antagonism had minimal effects on SMC differentiation. However, in both
Jak2
and
Tet2
models non-SMC derived fibroblast like cells expanded more than two-fold following IL-1β inhibition and were primarily enriched for matrix genes like
Col1a1
,
Col1a2
,
Lum
, and
Dcn
. Histological analysis of the fibroblast enriched protein Decorin revealed increased expression and localization in fibrous cap region. Together these findings suggest that IL-1β inhibition may promotes cap thickening by promoting fibroblast like cell accumulation in lesions. These results provide a potential mechanistic explanation for why IL-1β therapy was beneficial in CANTOS.
Abstract 110: The Histone Methyltransferase Setdb2 Modulates Macrophage TIMP-MMP Activity During Abdominal Aortic Aneurysm Development
Authors: Davis Frank , Melvin William , Mangum Kevin , Joshi Amrita , Wasikowski Rachael , Xing Xianying , Tsoi Liam , Gudjonsson Johann , Gallagher Katherine A
Published: October 2023

Introduction:
Abdominal aortic aneurysms (AAA) are characterized by aortic wall degradation leading to the fatal consequence of aortic rupture. Aortic remodeling is driven by an inflammatory macrophage (MΦ) phenotype and imbalance of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs). The epigenetic enzyme, SETDB2, plays a critical role in establishing MΦ phenotype and regulating gene expression. We examine the impact of SETDB2 on AAA development and establish SETDB2 as a novel target in the treatment AAAs.

Methods:
Single-cell sequencing was conducted on human AAA and control tissue samples. For our murine model, C57BL/6 mice were injected with an AAV encoding a PCSK9 gain-of-function mutation and then fed a saturated fat-enriched diet and infused with AngII (1 μg/kg/min) or saline. AAA maximum diameters were quantified and MΦs were sorted.
Setdb
and
Timp1-3
gene expression was examined. Chromatin immunoprecipitation (ChIP) was used to evaluate histone 3 lysine 9 trimethylation (H3K9me3).

Results:
Using single-cell RNA sequencing, we identified SETDB2 was profoundly upregulated in human aortic MΦs and two murine AAA models. Mechanistically, interferon-β regulates
Setdb2
expression via JAK/STAT signaling which trimethylates histone 3 lysine 9 (H3K9) at NF-κB binding sites on the TIMP1-3 gene promoters thereby suppressing
Timp1-3
transcription and leading to unregulated MMP activity. In vivo MΦ-specific knockout of SETDB2 (

Setdb2
f/f
Lyz2
Cre+

) inhibited AAA formation with significant suppression of vascular inflammation, MΦ infiltration, and elastin fragmentation. Gene expression and ChIP analysis demonstrated genetic depletion of SETDB2 prevented AAA development with removal of the repressive H3K9 trimethylation epigenetic mark on the
Timp1-3
promoter resulting in increased
Timp
expression and preserved aortic architecture. Lastly, pharmacological inhibition of the JAK/STAT pathway with a FDA approved inhibitor, Tofacitinib, limited
Setdb2
expression and inhibited AAA development.

Conclusions:
Taken together, our results identify SETDB2 as a critical regulator of MΦ-mediated inflammation and protease production in AAAs and suggest SETDB2 as a mechanistic target for the management of AAAs.
Abstract 112: Pcsk6 Ablation In Vascular Smooth Muscle Cells Promotes Abdominal Aortic Aneurysm Development And Rupture
Authors: Jin Hong , Lindquist Liljeqvist Moritz , Wu Zhiyuan , Micali Linda , Suur Bianca E , Lengquist Mariette , Zhang Xiang , Kronqvist Malin , Pauli Jessica , Hedin Ulf , Roy Joy , Matic Ljubica
Published: October 2023

Background and Aim:
We have previously shown that Proprotein Convertase Subtilisin/Kexin type 6 (Pcsk6) is strongly enriched in atherosclerotic plaques and plays a key role in lesion vulnerability and vessel wall healing by regulating vascular smooth muscle cell (VSMC) migration, proliferation, and extracellular matrix remodeling. Considering the broad importance of these processes in vascular pathologies, this study aimed to investigate the expression and function of Pcsk6 in the abdominal aortic aneurysm (AAA).

Methods and Results:
Bioinformatic analysis of human transcriptome microarray and RNA-sequencing data from human AAA biobanks of the Karolinska Institute and the Technical University of Munich have demonstrated a significantly increased level of Pcsk6 transcript in AAA tissues compared to controls, and even higher levels in ruptured AAAs. Single-cell RNA sequencing data demonstrated a significantly increased level of Pcsk6 transcript in fibroblasts and T cells but decreased in VSMCs. To further clarify if Pcsk6 dysregulation in VSMCs is a pathological reaction or if it has a causal role in AAA development and aortic dissection, we performed pilot animal studies: 1) peri-adventitial porcine pancreatic elastase (PPE) induced AAA model on male VSMC conditional Tagln-Pcsk6 deficient and control mice; 2) AngII infusion model on male Tagln-Pcsk6 deficient and control mice with ApoE-/- background. Interestingly, although no difference in baseline blood pressure, aortic dilatation, and constriction was found between Tagln-Pcsk6 deficient and control mice, ultrasound imaging and histology analyses showed that mice with Pcsk6 deficiency in VSMCs manifested remarkably larger AAA, as well as significantly increased rupture and dissection rate. Mechanistically, RNA sequencing analysis indicated significant inhibition of extracellular matrix components and biosynthesis signaling in aortas from Tagln-Pcsk6 deficient mice.

Conclusion:
PCSK6 expressed by VSMCs plays a substantial role in AAA progression and aortic dissection, opening new avenues for the therapy of aortic disease. Further mechanistic studies are necessary before this translational aspect can be evaluated.
Abstract 116: Variation In Lipoprotein (a) Level Reduction With Potent Lipid Lowering: An Analysis Of The Cholesterol Reduction And Residual Risk In Diabetes Study
Authors: Akinlonu Adedoyin , Boffa Michael , Lyu Chen , Zhong Hua , Jindal Manila , Bredefeld Cindy , Newman Jonathan D , Fisher Edward , Koschinsky Marlys L , Goldberg Ira J , Berger Jeffrey
Published: October 2023

Introduction:
Lipoprotein (a) [Lp(a)] is a recognized risk factor for ASCVD and a driver of residual risk. Proprotein Convertase Subtilisin/Kexin Type 9 inhibitors (PCSK9i) modestly decrease Lp(a) by ≈ 15%. However, there is significant heterogeneity in Lp(a) lowering response to PCSK9i. We hypothesize that the apolipoprotein a [apo(a)] isoform size is responsible for the variable response in Lp(a) lowering.

Methods:
We examined the change in Lp(a) levels from baseline in the Cholesterol Reduction and Residual Risk (CHORD) study. Participants with and without diabetes—all with LDL-C >100mg/dl—received potent lipid lowering therapy (LLT) for 30 days with (1) Evolocumab 140mg q 14 days, and (2) atorvastatin 80mg/day (or ezetimibe 10mg/day if already on a statin or statin intolerant). Lp(a) level was measured in the clinical lab. The apo(a) isoform size was measured by denaturing agarose gel electrophoresis followed by western blotting.

Results:
Among 87 participants (median age 53, 59% [51 of 87] female, 54% [47 of 87] diabetes), median Lp(a) was 35 mg/dl at baseline and 25 mg/dl at 30 days (P<0.01). Among 47 participants with a high Lp(a) at baseline (>30mg/dl), there was an average 14% reduction in Lp(a) from baseline to 30 days with considerable heterogeneity in response (Figure 1A). While baseline Lp(a) Level was not correlated with change in Lp(a) (Figure 1B), there was an inverse correlation between major isoform size and Lp(a) reduction (r= -0.64, P<0.001) (Figure 1C). After adjustment for age, sex, race/ethnicity, diabetes, and type of LLT, major isoform size was significantly associated with change in Lp(a) (Beta 0.96, 95% CI 0.94 to 0.98, P<0.001). An apo(a) isoform size of 18 discriminated the response of Lp(a) levels to LLT (isoform size ≥18 - 20% [14 to 26] reduction, isoform <18 - 5% [-8 to 19] increase).

Conclusion:
Our data indicate considerable variation in Lp(a) reduction with potent LLT, and that apo(a) isoform size is strongly associated with this variability.
Abstract 118: Positive Adipose Liver Crosstalk Protects Adipose MTP Knockout Mice from Hepatic Steatosis on an Obesogenic Diet
Authors: Rajan Sujith , Verano Michael , Aleman Jose O , Hussain M M
Published: October 2023

Introduction:
Our previous study demonstrated that adipocyte MTP regulates intracellular lipolysis by inhibiting ATGL activity. Adipose-specific MTP knockout mice (A-

Mttp
-/-

) had smaller adipocytes and showed increased thermogenesis and gained less weight on an obesogenic diet. They also exhibited moderately high plasma triglyceride levels and less hepatic steatosis compared to wild-type counterparts. In this study, we have elucidated the mechanism behind less hepatic lipid accumulation and moderately high plasma TG in A-

Mttp
-/-

mice.

Methods:
We characterized lipoproteins and studied hepatic ApoB production. We have analyzed adipose-liver crosstalk using two strategies 1) lipidomics of adipose tissue, plasma, and liver. 2) adipokine profiling. Further, we have used western blotting and qRT-PCR to check signaling and gene expression.

Results:
Plasma lipid profiling revealed increased TG content in the VLDL and LDL fraction of A-

Mttp
-/-

mice compared to

Mttp
f/f

mice. Additionally, A-

Mttp
-/-

mice showed significantly increased hepatic triglyceride production compared to the wild type. Lipidomic analysis revealed the movement of fatty acids from adipose tissue to the liver, resulting in significantly higher amounts of oleate, palmitate, linoleate, and stearate in the liver of A-

Mttp
-/-

mice compared to

Mttp
f/f

mice. A-

Mttp
-/-

mice liver also showed significantly increased expression of genes involved in fatty acid uptake and utilization such as
Cd36
,
Mcad
,
Fatp2
, and
Cpt1a
compared to

Mttp
f/f

mice. Further adipokine profiling showed a significantly higher amount of adiponectin and decreased amount of proinflammatory cytokines such as leptin in the plasma of A-

Mttp
-/-

mice compared to

Mttp
f/f

mice. We also found decreased ceramide levels and significantly increased phosphorylation of AMPK in the liver of A-

Mttp
-/-

mice compared to

Mttp
f/f

mice.

Conclusion:
We conclude that increased availability of substrate for TG production might contribute to increased ApoB secretion by the liver of A-

Mttp
-/-

mice. In addition, positive cytokine profile and increased adiponectin signaling in the liver of A-

Mttp
-/-

mice protects it from hepatic steatosis.
Abstract 119: Aging-related Activation Of Platelet Mtor Promotes Platelet Hyperreactivity And Thrombosis
Authors: Portier Irina , Manne Bhanu Kanth , Kosaka Yasuhiro , Denorme Frederik , Babur Ozgun , Wilmarth Phillip A , Aslan Joseph E , Rondina Matthew T , Campbell Robert
Published: October 2023

Background:
Aging is an independent risk factor for the development of cardiovascular disease. While aging is associated with increased thrombotic risk, mechanisms of platelet hyperactivation in aging remain undescribed.

Aims:
We examined the activation of intracellular signaling pathways in platelets from young and old human subjects and mice, where the mammalian target of rapamycin (mTOR) is hypothesized to be a mechanistic regulator of platelet hyperreactivity in aging.

Methods:
Platelets were isolated from healthy young (<45y) and older (>70y) human subjects. Platelet signaling was examined with quantitative mass spectrometry and Western blot. Platelet function was assessed with aggregometry. Platelet activation was also assessed in young (8-12 weeks) and aged (>18 months) wild-type (WT; mTOR
fl/fl
) and platelet-specific mTOR knockout mice (KO; mTOR
fl/fl
-Pf4-Cre).

Results:
We observed significant changes in phosphorylation of substrates within and downstream of mTOR/S6K and PI3K/Akt signaling pathways in aged humans compared to young. Western blot analysis confirmed significant (p<0.01) basal activation of the mTOR pathway, including P-AKT, P-p70S6K and P-4EBP1 in both aged humans and mice. Additionally, aged human and murine platelets had significant greater aggregation compared to young controls. Inhibition of mTOR either with torin-1 in aged humans or genetic deletion in aged mice reversed platelet hyperreactivity. In a collagen-epinephrine thrombosis model, aged WT mice succumbed significantly (p<0.001) faster compared to young WT mice, while aged KO mice had similar times to death as young WT mice. Mechanistically, we observed an increase in RAC-GTP (p<0.0001) downstream of mTOR activation in aged WT mice. In addition, we observed increased mitochondrial reactive oxygen species (mtROS) in resting platelets in aged WT mice compared to young WT and young or aged KO mice (p<0.05). Increased ROS led to significantly more P-p38 and thromboxane generation in aged WT mice (p<0.01). P-p38 and thromboxane generation were unchanged in aged and young KO mice.

Conclusions:
Aging is associated with increased platelet mTOR activation and platelet hyperreactivity mediated through increased activation of p38 and thromboxane generation.