bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved cluster to normal ratio protocol for increasing the lifetime of wireless sensor networks
Authors: Martey Abraham S. , Esenogho Ebenezer
Published: May 2022

The wireless sensor network (WSN) is a decentralized network that allows sensor nodes to freely join and quit the network, making it a unique network type that varies from centralized systems in which the fusion center decides on entry and exit. However, one of the key drawbacks of these networks is that the sensor nodes are small and located in remote areas. As a result, energy usage must be efficiently managed. In order to limit energy consumption, an efficient clustering protocol approach is necessary, which may be performed by dividing the networks into clusters and selecting cluster heads depending on the remaining energy and sensor distance. The energy hole problem has an impact on the performance of an energy-efficient technique. As a result, this work aims to extend the lifespan of WSNs, the goal of this research is to make the cluster to normal ratio (CTNR) Protocol better. Data forwarding nodes are added to the CTNR protocol hierarchy to enable hopping and avoid consuming nodes in order to solve the energy hole problem (sink). The proposed method is implemented in Matlab, and the results are compared to literature on energy usage and the amount of dead and living nodes. In terms of the performance indicators given, the results reveal that our proposed scheme beat others.
Modeling the impact of jamming attacks in the internet of things
Authors: Kerrakchou Imane , Chadli Sara , Ayachi Yassine , Saber Mohammed
Published: May 2022

Security is a key requirement in the context of the Internet of Things. The IoT is connecting many objects together via wireless and wired connections with the goal of allowing ubiquitous interaction, where all components may communicate with others without constraints. The wireless sensor network is one of the most essential elements of IoT concepts. Because of their unattended and radio-shared nature for communication, security is becoming an important issue. Wireless sensor nodes are susceptible to different types of attacks. Such attacks can be carried out in several various ways. One of the most commonly utilized methods is Jamming. However, there are also some other attack types that we need to be aware of, such as Tampering, Wormhole, etc. In this paper, we have provided an analysis of the layered IoT architecture. A detailed study of different types of Jamming attacks, in a wireless sensor network, is presented. The packet loss rate, energy consumption, etc. are calculated, and the performance analysis of the WSN system is achieved. The protocol chosen to evaluate the performance of the WSN is the S-MAC protocol. Different simulations are realized to evaluate the performance of a network attacked by the different types of Jamming attacks.
Cuckoo search algorithm and particle swarm optimization based maximum power point tracking techniques
Authors: Abdulaziz Sally , Atlam Galal , Zaki Gomaa , Nabil Essam
Published: May 2022

Increasing the efficiency of photovoltaic (PV) systems is a pressing issue, and several studies have focused on the Maximum Power Point Tracking (MPPT) techniques to extract the maximum PV output power. Many MPPT techniques have been discussed in the last decade, and optimization-based MPPT techniques have shown better performance than other MPPT techniques. In this study, two optimization techniques, the cuckoo search algorithm and particle swa9rm optimization with changing inertia weight techniques are discussed and applied to a PV system to track the maximum power point. The MSX-60 PV module and boost DC-DC converter are used in this paper to simulate and model the MPPT system using MATLAB/Simulink to show which technique has the best performance under various solar irradiation scenarios. In addition, different structures of PV arrays such as series-parallel, bridge link, and total cross-tied PV structures are simulated to analyze their effect on the efficiency of MPPT processes.
Modeling phasmophobia (fear of ghosts) using electroencephalogram
Authors: Izwan Safie Sairul , Khalid Puteri Zarina M. , Aimullah Mohd
Published: May 2022

Extreme fears towards ghosts and entities are defined as phasmaphobia. Those diagnosed with phasmophobia symptoms should control their own fears to avoid phasmaphobia attack. In this work, we present the development of phasmophobia detection electroencephalogram database (PDED).PDEDconsistsofanaverageof45minutes electroencephalography (EEG) recordings from eight electrodes situated on the frontal lobe of the brain area. A real-time fear assessment was conducted simultaneously with the EEG recording by the participant. Five different stimuli were used to induce fear in our experiment. 599 EEG epochs related to fear were extracted based on the timestamp recorded by each individual. Asymmetry relation ratio (ARR) techniques were used on these EEG to detect the presence of fear. The quality of long duration of EEG recording from PDED in recognizing fear was thoroughly presented based on ARR. In this study, 91.5% of fear emotion managed to be detected from these epochs. Using PDED, it is also proven that the changes of ARR reflected positive correlation towards the changes of the level of fear. Analysis using emotion recognition rate (ERR) curves indicated that, two electrodes, namely F7 and F8, were sufficient to recognized 88% of fear from the recordings.
An approach to analysis of arabic text documents into text lines, words, and characters
Authors: Abdo Hakim A. , Abdu Ahmed , Manza Ramesh , Bawiskar Shobha
Published: May 2022

Text line extraction from a text document image and segmenting it into isolate words and segmenting these words into individual characters are considered as one of the most critical processes in OCR systems development and turning the document into a searchable electronic representation, this paper presents a new approach to analyze the Arabic text documents, the proposed approach contains four steps, preprocessing, text line segmentation, word segmentation, character segmentation. The horizontal projection method are used to detect and extract the text line from preprocessed text documents image, in word segmentation step The space threshold are computed to determine the spaces among connected components in text line as within-word space or between-words space for segmenting the text line into isolate words, finally thinning method applied to find the skeleton of segmented word and analyses geometric characteristics of the characters to detect ligatures and characters. The proposed approach was tested and evaluated on a set of 115 text images, this set contains images from the KFUPM Handwritten Arabic TexT (KHATT) database and some images produced by the authors. The experiment results are extremely encouraging, with a success rate of 98.6% for lines segmentation, 96% for words segmentation, and 87.1% for characters segmentation.
Entre sucuris e queixadas: transformações nos mitos pano de origem da ayahuasca
Authors: Matos Marcos de Almeida
Published: May 2022

O artigo propõe examinar dois dos principais mitos pano relacionados à ayahuasca como transformações de uma série de outros mitos. Estabelecendo assim um conjunto de transformação, procura-se demonstrar que esses mitos estão intimamente ligados aos mitos que versam sobre a relação dos humanos com bandos de queixadas. Essa ligação pode ser compreendida à luz de uma hipótese bastante influente acerca da história da difusão ayahuasca, expandindo-a para a região dos altos rios Juruá e Purus. Em particular, trata-se de indicar algumas ideias para pensar como os mitos se transformaram para acomodar em seu seio o uso da ayahuasca, e o que essa mudança significa do ponto das cosmologias que produziram aqueles mitos.
A humanidade e seu(s) gênero(s): mito, parentesco e diferença no noroeste amazônico
Authors: Andrello Geraldo , Vianna João
Published: May 2022

Este artigo pretende articular duas investigações etnográficas, entre os Tukano e Baniwa, que abordam a relação entre parentesco e mito. Pretende-se compreender a produção das diferenças sociocosmológicas no Noroeste Amazônico recorrendo-se às narrativas míticas da origem da humanidade. A análise seguirá os acontecimentos que se desdobram a partir de Jurupari, uma criança extraordinária e artefato cerimonial possuidor de capacidades reprodutivas; da cobra-canoa, um animal-objeto-útero que gesta a humanidade; dos nascimentos nas cachoeiras de Hipana e Ipanoré; do adultério com uma cobra-homem branco; e da guerra entre homens e mulheres. Todos estes eventos, personagens e relações, articulam de modo complexo transespecificidade e relações de sexo cruzado. Esperamos ao final evidenciar esses dois distintos modos pelos quais a humanidade atual é estabilizada a partir de um fundo virtual de alteridade, notando as importantes transformações tukano e baniwa no Alto Rio Negro.
A cultura dos mitos: do regime de historicidade Karajá e sua potência “fria"
Authors: Nunes Eduardo Soares
Published: May 2022

Esse artigo toma a caracterização de Lévi-Strauss das “sociedades frias” como inspiração para discutir o regime de historicidade karajá, um povo do Brasil Central. Sua mitologia narra um tempo da criação, onde diversos aspectos do mundo são criados e a humanidade adquire os conhecimentos que hoje a caracterizam enquanto tal. Já a ação no mundo atual não é criadora: antes, as ações fazem aparecer formas já estavam lá, desde o princípio, pois inscritas pelo mito. Dessa maneira, as ações atuais fazem aparecer ou uma continuidade em relação aos tempos antigos, pois elicitam os mesmos nexos relacionais, ou uma ruptura marcada para com “a cultura”. Em um caso como no outro, o que é colocado em evidência é a potência “fria” de seu regime de historicidade, ou seu caráter contrahistórico. Para dar conta desse regime, escapando da ideia história formulada em termos de mudança e continuidade, argumento que é preciso separar o problema da história do problema do tempo.
A História de Carolina. Um estudo de caso sobre a difusão de um mito no Sudoeste Amazônico
Authors: Gow Peter
Published: May 2022

Um povo pode desaparecer, mas não uma história. Isso foi o que ocorreu com um mito kuniba, narrado a Nimuendaju por Carolina, em uma situação de remoção forçada promovida pelo Estado brasileiro no começo do século XX. O mito em questão falava da origem da lua, ocasionada pelo incesto de um irmão com uma irmã. Até o começo do século XX, povos vizinhos dos Kuniba, como os Cashinahua e os Kanamari, tinham narrativas muito distintas sobre a origem da lua. No entanto, ao longo do século, com a partida dos Kuniba, o tema do incesto entre irmão e irmã passa a estar presente nas narrativas, difundindo-se pela região das bacias dos rios Juruá e Purus. Isso conduz a uma reflexão sobre o problema da “causalidade informacional”, sobre a “política externa” estabelecida entre diferentes povos e sobre a “dialética espacial” promovida pelos mitos.
Apresentação
Authors: Vianna João , Soares-Pinto Nicole
Published: May 2022

O conjunto de trabalhos aqui reunidos é dedicado à relação entre mitologia e etnografia, partindo do estudo dos mitos e suas transformações em diversos povos indígenas de distintas regiões nas Américas indígenas. As pesquisas contempladas no dossiê percorrem um arco desenhado entre o Brasil Central, os afluentes do Rio Guaporé e o interflúvio Juruá-Purus no Sudoeste amazônico/Amazônia Ocidental, o Alto Rio Negro no Noroeste Amazônico, Mesoamérica e Noroeste Mexicano. Todos os artigos tomam como inspiração as lições das Mitológicas, de Claude Lévi-Strauss, uma das obras mais monumentais já produzidas no âmbito da antropologia e das ciências humanas. O impacto deste trabalho é, desde sua publicação inaugural em 1964, muito significativo para toda a pesquisa etnográfica realizada na etnologia indígena. Mas a despeito desta trivial avaliação, por sua complexidade, as Mitológicas estão longe de ter sido esgotadas. Este dossiê pretende adensar o estudo dos mitos entre povos indígenas, contribuindo para o entendimento de sua atualidade.
Abstract 107: PKM2 Promotes Neutrophil Activation And Cerebral Thrombo-inflammation: Therapeutic Implications For Ischemic Stroke
Authors: Dhanesha Nirav , Patel Rakeshkumar , Jain Manish , Kumskova Mariia , Thedens Daniel , Olalde Heena M , Nayak Manasa , Doddapattar Prakash , Leira Enrique C. C , Chauhan Anil K
Published: May 2022

In recent years compelling evidence has emerged that implicates role of metabolic reprogramming in the modulation of thrombosis and inflammation. The dimeric form of the metabolic enzyme pyruvate kinase muscle 2 (PKM2) enters nucleus and exerts protein kinase activity. We found that nuclear PKM2 levels were upregulated in neutrophils after the onset of ischemic stroke both in humans and in mice. Therefore, we evaluated the role of PKM2 in promoting thrombo-inflammation and ischemic brain injury.We generated novel myeloid cell-specific

PKM2
-/-

mice on wild-type (

PKM2
fl/fl
LysMCre
+

) and hyperlipidemic background (

PKM2
fl/fl
LysMCre
+
Apoe
-/-

). We observed that genetic deletion of PKM2 in myeloid cells limited inflammatory response in peripheral neutrophils and reduced neutrophil extracellular traps following cerebral ischemia/reperfusion. In the filament and autologous clot/rtPA stroke models, irrespective of sex, deletion of PKM2 in myeloid cells either in wild-type or hyperlipidemic mice reduced infarcts and enhanced long-term sensorimotor recovery. Laser speckle imaging revealed improved regional cerebral blood flow in myeloid cell-specific PKM2-deficient mice that was concomitant with reduced post-ischemic cerebral thrombo-inflammation (intracerebral fibrin(ogen), platelet (CD41- positive) deposition, neutrophil infiltration, and inflammatory cytokines). Mechanistically, PKM2 regulates post-ischemic inflammation in peripheral neutrophils by promoting STAT3 phosphorylation. Utilizing small molecule inhibitor (ML265) that inhibits PKM2 dimerization, we evaluated ex vivo thrombosis using human whole blood in a microfluidic flow chamber system. We observed a fivefold reduction in the thrombus growth rate in ML265-treated group. Similarly, ML265 treatment in mice resulted into significantly reduced poststroke neutrophil hyperactivation and improved short-term and long-term functional outcomes following stroke. Collectively, these findings identify PKM2 as a novel therapeutic target to improve brain salvage and recovery following reperfusion.
Abstract 117: Effect Of Home-Based Leg Heat Therapy On Walking Performance In Symptomatic Peripheral Artery Disease: A Pilot Randomized Clinical Trial
Authors: Monroe Jacob , Perkins Susan , Han Yan , Motaganahalli Raghunandan , Roseguini Bruno
Published: May 2022

Introduction:
Few non-invasive therapies currently exist to improve functional performance and restore quality of life in people with lower extremity peripheral artery disease (PAD). We previously demonstrated that supervised leg heat therapy using tube-lined trousers perfused with warm water elicited a clinically meaningful improvement in perceived physical functioning in patients with PAD.

Hypothesis:
We tested the hypothesis that unsupervised, home-based leg HT is safe and well-tolerated and improves walking performance in patients with symptomatic PAD.

Methods:
Thirty-four participants with an ankle-brachial index (ABI) values < 0.90 and a history of claudication were randomized into one of two groups: those receiving leg HT (n=18) or those receiving a sham treatment (n=16). Patients in both groups were provided with identical water-circulating trousers and a portable heating pump and were asked to apply the therapy daily (7 days/week, 90 min per session) for 8 consecutive weeks. The pump given to participants in the HT group circulated water at 43°C through the trousers, while in the sham group the pump circulated water at 33°C. The primary study outcome was the change in 6-min walk distance from baseline to 8 weeks.

Results:
Among the thirty-four patients enrolled in the study, 3 were excluded for failing to comply with study procedures and 1 voluntarily withdrew. One patient in the sham-treated group could not complete the 8-week assessment due to the COVID-19 pandemic. Overall, participants completed 96±4% of the required treatment sessions (Control: 97±4%, HT:96±3%). Further, no serious adverse reactions to treatment were observed. Changes in 6-minute walk distance between baseline and the 8-week follow-up were compared between groups using the Wilcoxon rank sum test, since the data distribution was non-normal. The change in 6-minute walk distance was significantly higher (p=0.029) in the group exposed to HT (n=15; median: 21.3; 25%,75% percentiles: 10.0,42.3) as compared to the control group (n=14; median: -0.91; 25%,75% percentiles: -5.7,14.3).

Conclusions:
These preliminary results suggest that home-based leg HT for 8 weeks promotes clinically meaningful changes in 6-minute walk distance in patients with symptomatic PAD.
Abstract 121: Exploiting Natural Genetic Variation In The Human Triglyceride Regulator
APOA5
To Understand Its Function
Authors: Stankov Sylvia , Vitali Cecilia , Park Joseph , Nguyen David , Englander S. Walter , Phillips Michael C , Hand Nicholas J , Rader Daniel J
Published: May 2022

Plasma triglycerides (TGs) are an independent predictor of the risk for CAD, the leading cause of death worldwide. TGs are also positively associated with risk and severity of hyperTG-induced acute pancreatitis (HTG-AP). Current therapies are often insufficient in reducing extremely elevated TGs. We believe that apolipoprotein A-V (apoA-V, encoded by APOA5) can fill this unmet medical need. ApoA-V is a potent modulator of TG metabolism; it enhances lipoprotein lipase TG hydrolysis. We hypothesize that naturally occurring human APOA5 variants can inform ApoA-V function and identify novel ApoA-V based therapeutic axes.

We used the Penn Medicine Biobank (PMBB) to identify and measure plasma TGs of carriers of APOA5 variants predicted to change ApoA-V structure-function. Then, we used hydrogen-deuterium exchange mass spectroscopy to determine the secondary structure of ApoA-V, thereby identifying putative functional domains onto which we mapped our variants of interest. Finally, we characterized the plasma lipid effects of these mutants using adeno-associated viral (AAV) vectors in apoa5 knockout (KO) mice.

We identified APOA5 variants associated with changes in plasma TGs. These variants primarily fall near the central heparin binding domain or C-terminal lipid binding domain. We selected APOA5 Q275X, which removes the entire lipid binding domain, for further interrogation. Apoa5 KO mice that received APOA5 Q275X AAV had higher plasma TGs than mice treated with WT APOA5 AAV. While WT ApoA-V protein associated with VLDL and HDL particles, Q275X ApoA-V protein appeared in lipoprotein free fractions.

We have identified APOA5 variants associated with plasma TG phenotypes in humans, and mapped them to an experimentally determined ApoA-V secondary structure to identify the functional domains likely impacted. We have identified APOA5 Q275X as a loss of function variant that fails to bind lipoprotein particles and is associated with elevated plasma TGs. Continued study of this and other interesting naturally occurring variants will provide insight into the function of ApoA-V in TG metabolism. These insights can help us to therapeutically enhance ApoA-V to rapidly reduce TG levels during acute HTG-AP and to help prevent recurrent HTG-AP.
Abstract 124: The Immunometabolic Role Of Platelets In Uncomplicated Malaria Infection
Authors: Blick-Nitko Sara K , Morrell Craig , Livada Alison C , Maurya Preeti , Ture Sara , Munger Joshua , Schafer Xenia
Published: May 2022

The malaria causing
Plasmodium
parasite is a major public health threat.
Plasmodium vivax (P vivax)
is the cause of uncomplicated malaria (UCM). Platelets are the cellular mediators of thrombosis and are also the most numerous immune cells in the blood, and a first responder to infections. Thrombocytopenia is a frequent complication of malaria, and a decrease in platelet count is a negative predictor of disease outcome. Malaria infection elicits a strong interferon gamma (IFNγ) response. IFNγ is a potent inducer of indoleamine 2,3-dioxygenase (IDO1) the rate-limiting enzyme that catalyzes the first step in Tryptophan (Trp) metabolism in the kynurenine (Kyn) pathway, shunting Trp away from serotonin production. Trp metabolism may be altered in malaria infection as a means to regulate immunometabolic responses, but the mechanisms remain unknown. Our platelet RNA-sequencing data from
P vivax
infected humans and from
P yoelii
infected mice showed increased expression of genes related to Trp metabolism, including
IDO1
. The role for platelets in metabolic pathway regulation is poorly explored in general, but particularly in infectious diseases. We introduce a novel idea that platelets participate in immunometabolism to infection. Using complementary experimental approaches such as liquid chromatography-mass spectrometry, ELISA, PCR, western blot, and flow cytometry, we test the hypothesis that platelets are a source of IDO1 in UCM malaria, and thrombocytopenia results in IDO1 depletion and immune dysregulation. We have discovered a role for platelets in Trp metabolic pathway regulation and that platelet regulated immune responses to malaria infection are in part dependent on the Trp metabolic pathways. During
P yoelii
infection there is a depletion of Trp, and increased Kyn metabolites, as well as decreased plasma serotonin. Platelet transfusions to infected mice can increase Kyn. Understanding the interplay between platelets and immunometabolic pathways may provide a better understanding of the impact of thrombocytopenia in diseases beyond malaria, and provide a means to improve malaria infection responses as well as improved platelet-directed therapeutics in many hematological, metabolic, and immune diseases
.
Abstract MP01: Retinoic Acid Receptor Alpha (Rarα) In Macrophages Protects From Diet-induced Atherosclerosis In Mice
Authors: Cassim Bawa Fathima Nafrisha , Zhu Yingdong , Gopoju Raja , Xu Yanyong , hu shuwei , Jadhav Kavita , Zhang Yanqiao
Published: May 2022

Atherosclerosis is a chronic vascular disease caused by inflammation and accumulation of lipids in the blood vessels. Macrophages in atherosclerotic lesions participate in lipid accumulation giving rise to formation of foam cells and production of mediating inflammatory cytokines. This makes them an attractive target for therapy. Previous studies have shown that retinoid signaling plays a broader role in modulating macrophage lipid metabolism and its inflammatory phenotype. Our preliminary results showed that when macrophages (RAW267.4 cells and mouse peritoneal macrophages) were treated with all-trans retinoic acid or retinoic acid receptor alpha (Rarα)-specific agonist, cholesterol efflux genes like Abca1 And Abcg1 and anti-inflammatory gene like Arg-1 were significantly increased. We hypothesized that retinoic acid receptor alpha (Rarα) may play a role in macrophages to protect against diet-induced atherosclerosis. To test this hypothesis, we conducted in-vivo studies using macrophage-specific Rarα knockout mice. Rarα floxed mice were crossed with lysozyme-cre transgenic mice to generate macrophage specific knockout mice (m-Rarα
-/-
). Rarα
fl/fl
(control) and m-Rarα
-/-
were given high fat/high cholesterol (HFHC) diet for 16 weeks. Intracellular triglyceride and cholesterol levels were significantly increased in peritoneal macrophages of m-Rarα
-/-
compared to the control. Analysis of macrophage gene expression showed increase in the expression of pro-inflammatory genes (Il-1β, Tnfα and Tgm-2) with decrease in genes involved in cholesterol efflux (Abca1 and Abcg1) and anti-inflammation (Arg-1 and Mrc-1) in m-Rarα
-/-
macrophages. This implies that loss of Rarα in macrophages can aggravate atherosclerosis. To test this hypothesis, we generated mice deficient in macrophage Rarα and low-density lipoprotein receptor and their control mice. These mice were then fed an HFHC diet for 16 weeks. The Oil Red O staining of aortas showed that mice lacking macrophage Rar had a significant increase in atherosclerotic plaques compared to the control mice. Our data demonstrate that Rarα in macrophages is protective from atherosclerosis.
Abstract MP04: Single Cell Profiling Identifies IgM To MDA-LDL Producing Human B Cells And Atheroprotective Role Of CD24
Authors: Pattarabanjird Tanyaporn , Nguyen Anh , McSkimming Chantel , Dinh Huy , Marshall Melissa , Ghosheh Yanal , Durant Chirstopher , Vallejo Jenifer , Gulati Rishab , Saigusa Ryosuke , Drago Fabrizio , Taylor Angela , Gonen Ayelet , Tsimikas Sotirios , Miller Yury I , Ley Klaus F , Hedrick Catherine C , McNamara Coleen A
Published: May 2022

Murine B-1 cells produce IgMs that inactivate oxidation-specific epitopes and protect against atherosclerosis. Despite compelling data that IgM to MDA-LDL (IgM
MDA-LDL
) is inversely associated with coronary artery disease (CAD) and cardiovascular (CV) events in human, the human B cell subtype that produces IgM
MDA-LDL
is unknown. Here, we utilized mass cytometry to identify the 11 circulating human B cell subtypes and discovered that only the frequency of CD20
+
CD27
+
IgM
+
cells was associated with IgM
MDA-LDL
levels. Notably, high expression of CD24, a GPI-anchored sialoglycoprotein, was associated with IgM
MDA-LDL
(r = 0.57, p = 0.002) and inversely associated with CAD severity (r = -0.58, p=0.001). Adoptive transfer (AT) of sorted CD20
+
CD27
+
IgM
+
CD24hi (B
27+IgM+CD24hi
) and CD24lo/- (B
27+IgM+CD24lo/-
) human B cells into humanized mice demonstrated that CD24 is critical for trafficking to the spleen and IgM production. Bulk RNAseq coupled with pathway analysis of B
27+IgM+CD24hi
and B
27+IgM+CD24lo/-
cells revealed enhanced BCR and CCR6 signaling in B
27+IgM+CD24hi
. Imaging flow cytometry indicated colocalization between CD24 and CCR6 which was abrogated by CD24 blocking antibody (CD24mAb). Trans-well migration and internalization assays demonstrated that CD24 blockade increased CCL20-induced CCR6 internalization (p < 0.05, n =4), and reduced CCL20-induced migration of B
27+IgM+CD24hi
cells (p < 0.05, n =4). CD24mAb treatment of B
27+IgM+CD24hi
cells prior to AT into humanized mice also reduced the amount of surface CCR6 on B
27+IgM+CD24hi
cells
in vivo
(p < 0.05, n = 4). Both CCR6 knockout and CD24 inhibition of B
27+IgM+CD24hi
cells reduced migration to the spleen (p < 0.05, n = 4), and plasma level of human IgM (p = 0.057, n = 4). Lastly, single cell multi-omics sequencing of PBMCs from 60 CAD subjects revealed elevated CCR6 and IgM signaling in B
27+IgM+CD24hi
cells in subjects with less severe CAD. Results provide the first evidence for a role for CD24 in regulating CCR6-mediated B cell homing to the spleen, IgM production and CAD in humans. Findings may have important implications for the new CD24 immunotherapy entering clinical trials and raise the possibility that augmenting CD24 and/or CCR6 on B
27+IgM+
cells may be an effective atheroprotective strategy.
Abstract MP11: Histone Methyl Transferase (suv39h1): Function In Smooth Muscle Cell Phenotypic Plasticity
Authors: Chatterjee Payel , Chakraborty Raja , Xie Yi , Sizer Ashley , Hwa John , Martin Kathleen A
Published: May 2022

Introduction:
The phenotypic plasticity of vascular smooth muscle cells (VSMCs) is central to growth and remodeling processes, but also contributes to cardiovascular disease. This unique ability of VSMCs to reversibly differentiate and de-differentiate is incompletely understood. SUV39H1 is a histone methyltransferase that specifically trimethylates Lysine-9 of histone H3 (H3K9me3), resulting in transcriptional repression through epigenetic gene silencing.

Hypothesis:
We hypothesized that SUV39H1 may play a role in SMC phenotypic switch.

Methods:
Using
in vitro
and
in vivo
approaches including knockdown, qPCR, western, and chromatin immunoprecipitation (ChIP) assays to determine role of SUV39H1 in SMC plasticity.

Results:
A qPCR array screen of epigenetic regulators in VSMCs identified SUV39H1 mRNA as upregulated with PDGF induced dedifferentiation and downregulated with rapamycin induced differentiation. This was confirmed at the protein level. SUV39H1 knockdown significantly increased VSMC contractile protein mRNA, protein levels and decreased dedifferentiation associated gene expression, and also decreased PDGF-induced VSMC migration (n=4). Interestingly, we found that expression of KLF4, the master transcriptional regulator of dedifferentiation in SMCs, was dramatically decreased after SUV39H1 knockdown (n=5). We further noted that SUV39H1 knockdown decreased KLF4 mRNA stability (n=3). Mechanistically, SUV39H1 knockdown increased miRNA143, a well-known repressor of KLF4. ChIP assays at contractile gene promoters showed a significant decrease in H3K9me3 mark and an increase in H3K27 acetylation, an activation mark (n=4). Carotid artery ligation induced intimal hyperplastic lesions in wild type C57BL/6 mice, showed a significant increase in SUV39H1 and H3K9me3 expression compared to uninjured vessels.

Conclusion:
We identify SUV39H1 an epigenetic regulator of VSMC phenotype whose expression and activity increase with dedifferentiation
in vitro
and
in vivo
. Mechanistically, SUV39H1 influences VSMC chromatin marks and KLF4 expression to repress contractile phenotype. Understanding the role of SUV39H1 and its targets may have implications for developing new therapeutic strategies for treating vascular diseases.
Abstract MP14: The Profibrotic Transition Of Vascular Smooth Muscle Cells-derived Resident Vascular Adventitial Progenitor Cells Contributes To Angiotensin II-induced Cardiac Fibrosis.
Authors: Lu Sizhao , Strand Keith , Jolly Austin , Dubner Allison , Mutryn Marie , Nemenoff Raphael A , Moulton Karen , McKinsey Timothy A , Weiser-Evans Mary C
Published: May 2022

Cardiovascular fibrosis is an important end-stage pathology that characterizes most cardiovascular diseases. Activated cardiac myofibroblasts (MFs) are the major contributors to ECM deposition in pathological fibrosis. However, due to potential heterogeneity of MFs, the origin of these cells remains controversial. Using highly specific smooth muscle cell lineage-tracing mouse models, we discovered the smooth muscle cell origin of a subpopulation of resident vascular adventitial progenitor cells, defined by the expression of the stem cell marker Sca1 (AdvSca1-SM cells), which rapidly proliferate and adopt a myofibroblast phenotype in response to acute vascular injury. Further, we identified a specific gene signature of active hedgehog/Wnt/β-catenin/Klf4 signaling in AdvSca1-SM cells and validated a Gli1-CreERT2-ROSA26-YFP (Gli1) reporter mouse model to be a faithful lineage tracing system for AdvSca1-SM cells. However, the function of AdvSca1-SM cells in cardiac fibrosis is unknown. Using immunofluorescent staining and label-free second harmonic generation (SHG) imaging, we observed the expansion and migration of AdvSca1-SM cells in close association with perivascular and interstitial cardiac fibrosis in a model of Angiotensin II (AngII)-induced cardiac fibrosis in Gli1 reporter mice. We performed single cell RNA sequencing (scRNA-seq) to examine the phenotype of AdvSca1-SM cells in this model. Our data showed that, upon AngII challenge, AdvSca1-SM cells differentiate along a profibrotic trajectory, which is characterized by loss of expression of
Klf4
, the lncRNA,
Meg3
, and stemness genes and up-regulation of myofibroblast genes. Importantly, AngII-induced profibrotic transcriptomic changes of AdvSca1-SM cells were recapitulated in human ventricular tissues exhibiting a gene signature of cardiac hypertrophy, emphasizing the translational significance of this phenotypic transition. Connectivity map analysis of the scRNA-seq data identified statins as potential candidates for inhibition of the profibrotic transition of AdvSca1-SM cells. In agreement, simvastatin induced the expression of stemness genes and inhibited TGFβ-induced up-regulation of αSMA and down-regulation of Klf4 in cultured AdvSca1-SM cells.
Abstract MP16: Multi-trait Gwas Of Atherosclerosis Detects Novel Loci And Potential Therapeutic Targets
Authors: Bone William P , Bellomo Tiffany , Chen Brian Y , Gawronski Katerina A , Zhang David , Park Joseph , Levin Michael , Tsao Noah , Klarin Derek , lynch julie , Assimes Themistocles L , Gaziano Michael , Wilson Peter , Cho Kelly , Vujkovic Marijana ,   the VA Million Veteran Program, O'Donnell Chris J , Chang Kyong-Mi , Tsao Philip S , Rader Daniel J , Ritchie Marylyn , Damrauer Scott , Voight Benjamin F
Published: May 2022

Atherosclerosis, which is the narrowing of the arterial walls via accumulation of cholesterol-rich arterial plaques, is the leading cause of vascular disease worldwide, including myocardial infarction and ischemic stroke. Although atherosclerosis affects arteries throughout the body, previous genome-wide association studies (GWAS) have been performed on specific atherosclerotic phenotypes such as coronary artery disease (CAD) and peripheral artery disease (PAD). There is substantial evidence to suggest that these more specific atherosclerosis phenotypes share a common genetic etiology. We performed a series of multi-trait GWAS using combinations of two atherosclerosis traits and seven atherosclerosis risk factor traits and detected 31 novel pleiotropic loci. We performed these multi-trait GWAS using the N-GWAMA multi-trait GWAS method and summary statistics for CAD (van der Harst et al. 2018), PAD (Klarin et al. 2019), body mass index (Pulit et al. 2019), type II diabetes (Vujkovic et al. 2020), smoking initiation (Wootton et al. 2020), and lipid traits (Klarin et al. 2019). We identified candidate causal genes for 14 of these loci through colocalization analysis with GTEx expression quantitative trait locus (eQTL) data.
VDAC2
and
PCSK6
are two candidate causal genes that our results and previous literature suggest are potential therapeutic targets.
VDAC2
eQTLs in aorta and tibial artery colocalized with a multi-trait GWAS signal detected in the CAD PAD multi-trait GWAS. Previous work has shown that
VDAC2
regulates apoptosis, and our results suggest increased
VDAC2
expression in smooth muscle cells could increase smooth muscle cell accumulation in atherosclerotic plaques. A sQTL (splicing QTL) for
PCSK6
in liver colocalized with a multi-trait GWAS signal between PAD and LDL. Further analysis of the sQTL signal suggested that the effect allele correlates with a more active isoform of
PCSK6
, which could increase lipid fractions and risk of atherosclerosis. These results show that joint analysis of atherosclerotic disease traits and their risk factors allows for identification of unified biology that may offer the opportunity for therapeutic manipulation.
Abstract MP20: Pd-1 Pathway Upregulation By Orchiectomy Attenuates The Aldosterone And High Salt Induced Aortic Aneurysms In Male Mice
Authors: Mu Xufang , Liu Shu , McClintock Timothy , Stromberg Arnold , Villasante-Tezanos Alejandro , Gong Ming C , Guo Zhenheng
Published: May 2022

Objective:
Male sex is a well-established risk factor for abdominal aortic aneurysms (AAA) but the underlying mechanisms remain to be fully understood. Using an aldosterone and high salt (Aldo/salt) induced AAA mouse model, we have demonstrated that androgen and its receptor mediate the high susceptibility to Aldo/salt induced AAA. The current study further investigates the mechanisms downstream of androgen.

Approaches and Results:
- To dissect the mechanisms connecting androgen and AAA, aortas were collected for RNA sequencing from 3 groups of 10-month-old wild-type mice #1 intact mice; #2 orchiectomized mice; and #3 orchiectomized mice plus DHT. All mice were given Aldo/salt for 7 days. Differentially expressed genes were analyzed using DESeq2 in R. We filtered genes that were upregulated in group #2 compared to group #1 and the up-regulation was reversed in group #3 by the DHT, or vice versa (fold change >1.5 and padj <0.05). Selected genes were run for gene ontology analysis in Enrichr (database Bioplanet 2019). Many pathways related to T cell activity were significantly enriched, particularly PD-1 signaling was one of the top pathways upregulated in orchiectomy group #2. PD-1 is known for its role as an immune checkpoint, and inflammation is a major hallmark for AAA development. Therefore to explore the role of PD-1, we first confirmed the PD-1 mRNA changes in aorta by qPCR. Secondly, IHC staining also showed PD-1 protein was significantly increased in the spleen of orchiectomized mice compared to intact controls. Finally, to investigate the potential causal role of PD-1 in the androgen-mediated aortic aneurysms formation, we injected αPD-1 antibody or control IgG antibody to orchiectomized mice 3 days before and during the 8 weeks of Aldo/salt administration. Results showed that 5 out of 12 αPD-1 mice, while none of the 8 control mice developed aortic aneurysms (p<0.05).

Conclusions:
PD-1 pathway is involved in the androgen associated high susceptibility of Aldo/salt induced aortic aneurysms in mice.
Abstract P105: Inducible Deletion Of Fibrillin-1 In Smooth Muscle Cells Has Modest Effects On Thoracic Aortic Dilatation In Adult Mice
Authors: Ito Sohei , Ohno Satoko , Franklin Michael , Moorleghen Jessica J , Howatt Deborah A , Sawada Hisashi , Lu Hong S , Daugherty Alan
Published: May 2022

Introduction:
Fibrillin-1 (
Fbn1
) mutations are associated with thoracic aortic aneurysms (TAAs) in patients with Marfan syndrome. Constitutive deletion of
Fbn1
leads to neonatal death due to aortic rupture in mice. However, the role of
Fbn1
in maintaining aortic integrity during the adult phase remains unclear. Since smooth muscle cell (SMC) is a major constituent of the aortic media, this study examined whether deletion of
Fbn1
in SMCs of adult mice leads to compromise of aortic integrity.

Methods and Results:
Fbn1
floxed mice were generated that included insertion of tdTomato as a reporter for gene deletion. Female
Fbn1
floxed mice were bred with male
Fbn1
floxed mice expressing CreER
T2
driven by an
Acta2
promotor. Tamoxifen was injected intraperitoneally for 5 days into male
Fbn1
floxed littermate mice that were either Cre negative or positive (n=6-8/genotype, 5-week-old) to create sm
Fbn1
+/+ and -/- mice, respectively. Gene recombination was confirmed by the presence of tdTomato in sm
Fbn1
-/-
mice using PCR and confocal microscopy. To investigate the impact of SMC-specific
Fbn1
deletion, male sm
Fbn1
+/+ and -/- littermates terminated at two intervals, 5 and 36 weeks after completion of tamoxifen injections. In situ aortic images were captured, and aortic diameters were measured at ascending, arch, and descending thoracic aortas. Five weeks after tamoxifen injection, aortic diameters of smFbn1+/+ and -/- mice were not different. Despite the long-term deletion of
Fbn1
in SMCs, aortic dilatation was not observed in any regions of aortas in sm
Fbn1
-/- mice compared to wild type littermates 36 weeks after tamoxifen injection. Next, we investigated whether
Fbn1
deletion in SMCs augmented angiotensin II (AngII)-induced aortic dilatation. AngII was infused subcutaneously into male sm
Fbn1
+/+ and -/- littermates for 12 weeks (n=11-12/genotype), started 2weeks after completion of tamoxifen injections. AngII-induced aortic dilatations were augmented modestly by SMC-specific
Fbn1
deletion (+/+ 1.7±0.0, -/- 1.9±0.1 mm, P=0.047).

Conclusion:
Deletion of
Fbn1
in SMCs did not cause spontaneous TAA formation and had modest effects on AngII-induced TAAs in adult mice.
Fbn1
in SMCs does not exert a critical role in mice after the aortic structure has formed.
Abstract P106: Sustained Inhibition Of High Mobility Group Box 1 Mrna By Antisense Oligonucleotides Demonstrated A Protracted Half-life Of The Protein
Authors: Mohammadmoradi Shayan , Sawada Hisashi , Ito Sohei , Ye Dien , Mullick Adam E , Howatt Deborah A , Franklin Michael , Lu Hong S , Daugherty Alan
Published: May 2022

Background and Objectives:
High-mobility group box 1 (HMGB1) is a highly conserved nonhistone DNA-binding nuclear protein which is present in almost all eukaryotic cells. HMGB1 is proposed to exert effects on several forms of vascular disease including atherosclerosis, pulmonary hypertension, and abdominal aortic aneurysm. However, the lack of a viable whole-body genetic deletion has increased interest in pharmacological approaches to sufficiently inhibit HMGB1 to investigate its role in pathological conditions. In the current study, we assessed the efficacy of a novel antisense oligonucleotides (ASOs) approach to deplete HMGB1 in mice.

Methods and Results:
Either ASOs (25 mg/kg/day) or phosphate-buffered saline (PBS) were injected intraperitoneally into male C57BL/6J mice (8-10-week-old). ASOs were injected at day 0 and day 3 in the initial week and once a week for the duration of the study. Mice were terminated at either 2, 4, or 12 weeks after the initial injection of ASOs (N = 5/group). Since HMGB1 is highly abundant in kidney and liver, mRNA and protein abundance of HMGB1 were examined subsequently in these organs by qPCR and Western blot analyses.ASO administration resulted in 90% decrease of
Hmgb1
mRNA abundance in kidney and liver by 2 weeks of ASO injection. The decreased
Hmbg1
mRNA abundance was observed consistently at both week 4 and 12. Surprisingly, ASOs failed to deplete HMGB1 protein abundance in kidney and liver at 2- and 4-week time intervals. However, HMGB1 protein abundance was markedly reduced to less than 20% in kidney and liver tissues at week 12, indicating an extended protein half-life.

Conclusion:
These findings reveal the protracted interval need to deplete HMGB1 protein depletion after mRNA inhibition.
Abstract P110: Molecular Modeling Of Apolipoprotein(a): Insight Into The Orientation Of Key Functional Domains Mediating Lipoprotein(a) Assembly And Ligand Binding
Authors: Daichendt Luke G , Szabla Robert , Junop Murray S , Koschinsky Marlys L , Boffa Michael B
Published: May 2022

The apo(a) component of Lp(a) plays a major role in mediating the pathological effects of Lp(a) in vascular disease. Apo(a) is composed of repeating kringle (K) domains, of which 10 types are unique (KIV1; KIV3-KIV10; KV) and one (KIV2) is present in variable numbers. KIV7 and KIV8 contain weak lysine binding sites (LBS) that play a role in Lp(a) assembly and KIV10 contains a strong LBS important for binding of Lp(a) to biological substrates and the attachment of pro-inflammatory oxidized phospholipids. Following the kringle domains is an inactive protease domain. While the structures of individual kringles have been solved, no structural information exists on intact apo(a). We therefore performed
in silico
molecular modeling of apo(a) KIV7-protease. This region contains the key functional domains of apo(a) and has a similar topology to plasminogen. The structures of human, rhesus monkey (lacking the KIV10 strong LBS) and baboon (lacking KV) apo(a) were modeled with Swiss-Model using the crystal structure of the closed conformation of full-length plasminogen (pdb: 4A5T) as the template, followed by energy minimization using Rosetta Relax. The lowest energy human apo(a) model represented a compact, plasminogen-like, tertiary structure that promoted high solvent exposure of the KIV7 LBS and the KIV9 unpaired cysteine, moderate exposure of the KIV10 LBS, and low exposure of the KIV8 LBS. Interestingly, there was substantial variability in the energy minimized models; of the 10 lowest energy models from Rosetta Relax, two showed notable translation of the protease domain resulting in increased solvent exposure of KIV8 at the expense of KIV10. The rhesus and baboon models were highly similar to human but showed much less variability in energy minimized models. Our findings show that not all key sites in apo(a) are highly solvent exposed, but the flexibility of human apo(a) structure suggests the possibility of conformational changes that could alter their accessibility.
Abstract P112: A Simple, Rapid, And Sensitive Fluorescence-based Method To Assess Triacylglycerol Hydrolase Activities
Authors: Rajan Sujith , De Guzman Hazel C , Palaia Thomas , Goldberg Ira J , Hussain M
Published: May 2022

Introduction:
Lipases constitute an important class of water-soluble enzymes that catalyze the hydrolysis of hydrophobic triacylglycerol (TAG). Their activity is usually measured after isolation and quantification of the hydrolyzed free fatty acids. We have used NBD-labeled lipids previously to develop TAG, cholesteryl ester, and phospholipid transfer assays for microsomal triglyceride transfer protein. To date, however, NBD-labeled TAG (NBD-TAG) has not been used to assess the enzymatic activities of different lipases.

Methods:
We developed methods to measure lipase activities using adipose triglyceride lipase (ATGL), lipoprotein lipase (LpL), and pancreatic triglyceride lipase (PNLIP) as model lipases. In these assays, we incubated a source of ATGL, LpL, or PNLIP with substrate emulsions containing nitrobenzoxadiazole (NBD)-labeled TAG in phosphatidylcholine (PC) and phosphatidylinositol (PI), measured increases in NBD fluorescence with time, and calculated enzyme activities.

Results:
Incorporation of NBD-TAG into PC vesicles resulted in some hydrolysis; but required product isolation step to measure NBD fluorescence. However, incorporation of PI into these NBD-TAG/PC vesicles significantly increased substrate hydrolysis in time, protein, and substrate concentration dependent manner negating the need to isolate products to measure lipase activities. Further, increasing the ratio of NBD-TAG to PC enhanced substrate hydrolysis. Next, we tested specific lipase inhibitors and found that orlistat inhibits all three enzymes indicating that it is a pan-lipase inhibitor.

Conclusions:
We describe a simple, rapid fluorescence-based TAG hydrolysis assay to assess three major TAG hydrolases: ATGL localized intracellularly, LpL localized at the extracellular endothelium, and PNLIP produced and secreted from the pancreas into the intestinal lumen. The major advantages of this method are its speed, simplicity, and avoidance of a product isolation step. This assay is potentially applicable to a wide range of lipases and is amenable to high-throughput screening to discover novel modulators of triacylglycerol hydrolases, and in the diagnosis of diseases associated with increases in plasma lipase activities.