bluuebunny/crossref_metadata_2025 huggingface dataset:
Uma arte exuberante e compromissada: Gori Muñoz em três tempos
Authors: Santos Margareth dos
Published: July 2021

Entrevista realizada com María Antonia Muñoz-Malajovich sobre a trajetória de Gori Muñoz.
Estruturas Antigas, Tectônica Recente: Diferentes Controles na Evolução do Relevo na Região das Altas Cristas Quartzíticas da Mantiqueira Meridional
Authors: Marques Neto Roberto , Da Silva Felipe Pacheco , Moreira Juliana Alves
Published: July 2021

As altas cristas da Mantiqueira Meridional figuram como estruturas festonadas pela rede de drenagem que demanda indiretamente o Oceano Atlântico, figurando como uma unidade morfoestrutural cuja geomorfogênese coaduna retração erosiva de escarpas a processos diastróficos morfotectônicos, encerrando um sistema geomorfológico de alta complexidade, onde os efeitos neotectônicos se sobrepõem às expressões geomorfológicas regionais forjadas em estruturas antigas reativadas durante o rifte Continental do Sudeste do Brasil. Com base em abordagens metodológicas voltadas para as interpretações morfoestruturais e morfotectônicas, a pesquisa aqui divulgada identificou importantes aspectos da tectônica ativa atuando na evolução do relevo regional, cuja espacialidade é estabelecida pelas superfícies de base, pela configuração da drenagem e padrão de lineamentos, pelo reconhecimento e mapeamento de feições morfotectônicas e interpretação de seu significado, bem como pela compartimentação da área segundo seu significado morfoestrutural e morfotectônico.
Um Estudo Sobre o Conceito de Natureza
Authors: Castro Daniel Stella
Published: July 2021

A partir da revisão bibliográfica de “A História da ideia de Natureza” de Robert Lenoble, “A Natureza”, de Maurice Merleau-Ponty e “The Concept of Nature” de Alfred Whitehead, o principal objetivo do presente estudo foi apresentar de maneira sintética as principais noções e conceitos de Natureza discutidos em tais obra, como: Natureza mágica; Natureza como finalidade; Natureza como exterioridade; Natureza como um sistema; Natureza humanista e; o natural na Natureza. Assim, se buscou demonstrar a dificuldade em traçar limites para uma definição única e objetiva sobre Natureza. Posto isso, concluiu-se que a Natureza isolada em diferentes aspectos é uma abstração científica eleita como método em diversos campos disciplinares na busca de entender um determinado fenômeno. Nesse sentido, postula-se que a maneira como um indivíduo ou um grupo social entende a Natureza tem relação direta com suas atitudes no mundo vivido, atitudes que são construídas culturalmente.
Ilhas de calor urbano em Montes Claros – MG
Authors: Almeida Maria Ivete Soares de , Silva Lucas Augusto Pereira da , Leite Marcos Esdras , Laurentino Carla Milena de Moura , Marques Samuel Carlos Santos , Leite Manoel Reinaldo
Published: July 2021

O objetivo desse trabalho é analisar a distribuição espacial das Ilhas de Calor na Cidade de Montes Claros – MG. A escolha dessa área se justifica pelo crescimento populacional e a expansão urbana que a mesma vem passando nos últimos anos. Para execução desse trabalho, dotou do uso de técnicas de sensoriamento remoto para modelagem térmica e índices de comprimentos de onda. Foram utilizados os produtos orbitais dos satélites Landsat 5 e 8 (sensores TM e OLI/TIRS, respectivamente) referentes às datas: 28/08/1985 e 18/09/2016. Os resultados mostraram que a distribuição espacial das Ilhas de Calor está relacionada as alterações na ocupação e uso do solo urbano, variando em escala espaço-temporal, apresentando intensificação nas regiões centrais para as anomalias positivas, e nas bordas apresentam aumento das distribuições de anomalias negativas, sendo essas correlatas ao incremento de áreas verdes em condomínios de alto padrão.  A elevada de temperatura de superfície possui relação proporcional com o NDBI e inversamente com o NDVI, considerando as características da cobertura do solo. Esse estudo se mostra como instrumento de planejamento urbano, sobretudo considerando os aspectos de uso e ocupação do solo para com efeitos no âmbito meteorológico e climatológico, haja vista que estes elementos são essenciais para o bem estar humano.
Vulnerabilidade costeira a partir da abordagem multicritério: estudo de caso no litoral sul do Espírito Santo
Authors: Filgueiras Gilberto Daniel Lima , Albino Jacqueline
Published: July 2021

Questões relacionadas à vulnerabilidade costeira importância ao longo de todo mundo, devido às questões climáticas, elevação do nível dos oceanos e expansão urbana sobre tais regiões. Faz-se, necessário entender o grau de vulnerabilidade costeira e os fatores envolvidos, com o intuito de evitar perdas socioeconômicas e também ambientais. O presente estudo objetivou analisar, mediante o Índice de Vulnerabilidade Costeira (CVI), de que modo a vulnerabilidade costeira é alterada com a mudança das características ao longo da costa dos municípios de Marataízes e Presidente Kennedy, localizados no litoral sul do estado do Espírito Santo. Geomorfologia costeira, Modelo digital de elevação, taxas de mobilidade costeira e distância da isóbata de 20m, foram obtidas por meio de dados de sensoriamento remoto. A espacialização e integração dos dados mediante o CVI foi efetuado com utilização de software GIS. Os resultados mostram que em 49% dos 44km do litoral a vulnerabilidade é moderada a muito alta, com ênfase para as regiões do litoral com existência de falésias e extensas planícies costeiras. Discute-se diferentes formas de ação para minimizar prejuízos frente ao processo de futura urbanização e as diferenças entre as ações a serem tomadas pelos municípios de Marataízes e Presidente Kennedy, que possuem padrões de urbanização contrastantes. Conclui-se que o conjunto de dados de sensoriamento remoto se mostrou eficaz na determinação da vulnerabilidade costeira e que as questões relacionadas às mudanças climáticas devem começar a serem enfrentadas sob a ótica de longo prazo.
A atualidade da teoria dos circuitos na compreensão das práticas econômicas populares: o caso da Economia Criativa em Araraquara/SP
Authors: Ramos Renan Augusto
Published: July 2021

As práticas econômicas alternativas estão em evidência no contexto brasileiro do século XXI. Constatamos, nesse sentido, a existência de feiras econômicas em Araraquara que reforçam a temática da economia urbana. Dessa maneira, sob o apoio de um governo municipal eleito em 2016 com o slogan “Cidade Solidária e Participativa”, é necessário que se construam iniciativas de compreensão do fenômeno da Economia Solidária e Criativa no município de Araraquara. Para tanto, recorremos à Geografia para fornecer arcabouço teórico-metodológico de referência ao entendimento da problemática. Em específico, foi utilizada a teoria dos circuitos econômicos de Milton Santos (2018), que constata, nos países subdesenvolvidos, a fragmentação da economia urbana em dois circuitos. A proposta, então, qualificou essas práticas econômicas como representativas do circuito inferior da economia urbana, que almeja fornecer ocupação nos circuitos gerais da produção de valores. A partir disso, com a realização de uma pesquisa empírica nas edições da “Rolêfeira”, aplicação de entrevistas e questionários com os sujeitos sociais de interesse, a finalidade do trabalho foi observar a importância desses empreendimentos enquanto estratégias que buscam superar o desemprego e promover o desenvolvimento econômico e social em Araraquara, bem como compreender a própria interface de comunicação estabelecida entre a sociedade civil e o governo municipal.
Relação entre uso e cobertura da terra e parâmetros biofísicos no Cerrado Brasileiro
Authors: Martins Alécio Perini , Galvani Emerson
Published: July 2021

O uso e cobertura da terra é fundamental na regulação do ciclo da água, influenciando o processo de evapotranspiração. Mudanças nas categorias de uso vão interferir diretamente no saldo de radiação por meio da alteração de três parâmetros biofísicos: o albedo, o índice de vegetação e a temperatura de superfície. Neste sentido, a pesquisa objetiva avaliar a relação entre o uso e cobertura da terra e parâmetros biofísicos envolvidos na estimativa de fluxos de calor e evapotranspiração real. Estes parâmetros foram estimados por meio do SEBAL (Surface Energy Balance Algorithms for Land), modelo essencialmente matemático que necessita de poucos dados coletados em superfície para sua calibração. A área de estudo compreende sete bacias hidrográficas localizadas em áreas significativas do Sudoeste de Goiás e Noroeste de Mato Grosso do Sul, domínio do Cerrado, com grande diversidade de ocupação das terras, onde predominam as áreas de produção de grãos e pecuária de corte. A temperatura de superfície foi identificada como o parâmetro biofísico com maior correlação com o saldo de radiação, fluxos de calor e evapotranspiração, apresentando coeficientes de determinação superiores a 80%. O albedo, apesar de apresentar coeficiente de determinação na faixa de 60%, também mostrou-se como importante indicativo de mudanças ambientais com efeito direto sobre o ciclo da água. A análise estatística indica ainda que o modelo mostra-se eficaz para estimativa de evapotranspiração em grandes áreas, com diversidade de uso e poucos pontos de monitoramento em campo.
Oral health indicators and sociodemographic factors in Brazil from 2008 to 2015
Authors: Santos Juliana Leandro dos , Ferreira Raquel Conceição , Amorim Leonardo de Paula , Santos Anna Rachel Soares , Chiari Antônio Paulo Gomes , Senna Maria Inês Barreiros
Published: July 2021

OBJECTIVE: To evaluate the annual variation of oral health and primary care coverage, the tooth extraction ratio, and the average of supervised toothbrushing in Brazilian municipalities according to social development and population size. METHODS: Public secondary data were analyzed. The outcomes were health service indicators (oral health coverage, primary health care coverage, tooth extraction ratio, and average of supervised tooth brushing) estimated for all Brazilian municipalities annually from 2008 to 2015. Mixed-effect multilevel regression models with random intercept and slopes were fitted with a cross-interaction term to estimate the annual percent variation according to the Municipal Human Development Index (MHDI) and population size. RESULTS: Municipalities with low MHDI presented an annual increase in oral health and primary care coverage of 2.65% and 2.23%, respectively, which was significantly higher than municipalities with medium and high MHDI. Oral health and primary care coverage were 69.26% and 35.00% lower among municipalities with a large population. Municipalities with medium and high MHDI showed an annual decrease in tooth extractions of 5.15% and 5.02%, respectively. An annual decrease was observed in the average of supervised toothbrushing of 9.81% and 4.57% in municipalities with low and medium MHDI, respectively. The tooth extraction ratio was higher among larger municipalities; the relation is inverse for supervised toothbrushing. CONCLUSIONS: The access to primary care and oral health services increased in Brazil, while a decrease occurred in mutilating treatment and provision of preventive actions, with disparities among municipalities with different MHDI levels over time.
Covid-19 no Nordeste do Brasil: primeiro ano de pandemia e incertezas que estão por vir
Authors: Kerr Ligia Regina Franco Sansigolo , Kendall Carl , Almeida Rosa Lívia Freitas de , Ichihara Maria Yury , Aquino Estela Maria L , Silva Antônio Augusto Moura da , Ximenes Ricardo Arraes de Alencar , Albuquerque Maria de Fatima Pessoa Militão de , Almeida-Filho Naomar , Souza Rafael Felipe , Brandão Filho Sinval Pinto , Souza Wayner Vieira de , Barreto Maurício Lima
Published: July 2021

OBJETIVO Analisar a epidemia da covid-19 na região Nordeste do Brasil, uma das mais atingidas por essa virose. MÉTODOS Os dados oficiais para covid-19 dos estados do Nordeste são referentes ao período de março de 2020 a março de 2021. A análise para capitais e estados da série do acumulado semanal de casos e de óbitos confirmados foi feita com emprego do aplicativo JoinPoint Trend Analysis. RESULTADOS Em um ano, o Nordeste acumulou 22,9% dos casos e 21,5% dos óbitos do país pela covid-19. No início da pandemia, todos os estados apresentaram um crescimento sustentável de casos, primeiro nas capitais e depois interior. Em seguida, observam-se decréscimos em todos os estados e suas capitais, porém muitos permanecem com números elevados. Em meados do segundo semestre de 2020, o número de casos começa a crescer nos estados e suas capitais, alguns em velocidade explosiva, em especial no final de 2020 e início de 2021. Padrão similar é observado para os óbitos, os quais ultrapassaram ou aproximam-se do teto observado na primeira onda, na qual todas as capitais e estados do Nordeste adotaram intensas medidas de isolamento. Fortaleza, Recife e Teresina atingiram os maiores índices de isolamento de todas as capitais, perto de 0,60. Esse índice decresce, com tendência leve de crescimento até final de dezembro, voltando a decrescer. Com exceção de Fortaleza e Salvador, as demais capitais caíram para menos de 0,40. CONCLUSÃO O Nordeste brasileiro e o país estão em uma situação sanitária, social e econômica cada vez mais complexa. É necessário acelerar o processo de vacinação e manter as medidas não farmacológicas – constante uso de máscaras faciais, medidas de distanciamento e cuidados de higiene –, além de políticas de proteção aos trabalhadores que perderam as suas rendas e subsídios aos pequenos empresários.
Abstract 113: Monocytes Embedded in Plasma Clots Synthesize Coagulation Factors: Balancing Procoagulant and Anticoagulant Signals
Authors: Campbell Robert A , Vieira de Abreu Adriana , Weyrich Andrew S
Published: July 2021

Objective:
Previous studies have demonstrated that vascular cells acutely regulate fibrin clot structure and stability. However, the mechanisms by which fibrin clots push vascular cells, in particular leukocytes, to synthesize pro-and anti-coagulant proteins is unknown. Here, we characterize monocyte-derived pro-and anti-coagulant activity in clots.

Methods:
Human monocytes were isolated from whole blood using CD14 microbeads. In parallel, cell-free plasma was isolated and clot formation induced in the presence of monocytes by the addition of tissue factor and calcium. RNA was isolated using RNAzol. Cell surface tissue factor (TF), tissue factor pathway inhibitor (TFPI), urokinase plasminogen activator (uPA) and its receptor (uPAR) as well as plasminogen activator inhibitor-1 (PAI-1) was measured by ELISA. TF activity was measured using a chromogenic factor Xa assay.

Results:
Monocytes embedded within plasma clots transcribed mRNA for uPAR (2.3-fold), uPA (9.5-fold), TF (9.2-fold), TFPI, and PAI-1 (6.2-fold) in a time-dependent manner. Consistent with this transcriptional response, increased levels of TF, TFPI, and uPAR protein were visually detected on the monocyte surface 18-hours after plasma clot formation. Accumulation of TF, TFPI and uPAR protein was blocked by actinomycin D and cycloheximide, indicating protein synthesis was dependent on transcription of new mRNA. TF-activity was similarly increased (8.9-fold) on the surface monocytes (i.e., increase in plasma-clot versus suspension monocytes). Lastly, active PAI-1 (11.1-fold increase), but not uPA, increased in the culture supernatant over time.

Conclusions:
Plasma clot formation induced robust increases in coagulation-related mRNAs and their corresponding proteins. The balance between the transcription of pro- and anti-coagulant mRNAs and their subsequent translation into bioactive protein likely plays key roles in clot development, stability and resolution.
Abstract 126: Lysophosphatidic Acid Induces Early Growth Response Protein 1 Expression via the Protein Kinase Cδ-Regulated ERK and JNK Activation in Vascular Smooth Muscle Cells
Authors: Iyota Takuya , Sun Longshen , Hao Feng , Schmitz-Peiffer Carsten , Xu Xuemin , Cui Mei-Zhen
Published: July 2021

Lysophosphatidic acid (LPA) modulates vascular cell function in vitro and in vivo via regulating the expression of specific genes. Previously we reported that a transcriptional mechanism controls LPA-induced expression of early growth response protein 1 (Egr-1) in vascular smooth muscle cells (SMCs). Egr-1 is a master transcription factor mediating the expression of various genes that have been implied to modulate a broad spectrum of vascular pathologies. In this study, we determined the essential intracellular signaling pathway leading to LPA-induced Egr-1 expression. Our data demonstrate that activation of ERK-1/2 and JNK, but not p38 MAPK, is required for LPA-induced Egr-1 expression in SMCs. We show the first evidence that MEK/ERK-mediated JNK activation leads to LPA-induced gene expression. Examining the upstream kinases that mediate ERK and JNK activation leading to Egr-1 expression, we found that LPA-induced activation of MAPKs and expression of Egr-1, are dependent on PKC activation. We observed that LPA rapidly activates PKC δ and θ. Overexpression of dominant negative PKCδ, but not dominant negative PKCθ, diminished activation of ERK and JNK and blocked LPA-induced expression of Egr-1 mRNA and protein. We have also evaluated LPA receptor involvement. Our data reveal an intracellular regulatory mechanism: LPA induction of Egr-1 expression is via the LPA cognate receptor, LPA1-dependent and PKCδ-mediated ERK and JNK activation. This study provides the first evidence that PKCδ mediates ERK and JNK activation in the LPA signaling pathway and that this pathway is required for LPA-induced gene regulation as evidenced by Egr-1 expression.
Abstract 136: Differential Effect of Nitric Oxide on the 26S Proteasome in Type 1 and Type 2 Diabetic Rats
Authors: Rodriguez Monica P
Published: July 2021

INTRODUCTION:
We have shown that nitric oxide (NO) is more effective at inhibiting neointimal hyperplasia in rodent models of type 2 versus type 1 diabetes. To explain this difference, we looked at the 26S proteasome and deubiquitinating enzyme, USP5, since recent data revealed altered protein ubiquitination in diabetic environments. We hypothesize that NO has a differential effect on the 26S proteasome, the proteasomal activator, PA28-α, and/or USP5 in type 1 vs. 2 diabetic rat models.

METHODS:
The carotid artery injury model was performed in non-diabetic (LZ), type 1 (streptozotocin-injected LZ [STZ]), and type 2 diabetic (ZDF) rats. Groups included control, injury, and injury+PROLI/NO [n=4-7/group]. At 72hrs, carotid arteries were homogenized. PA28-α and USP5 expression was assessed by Western blot analysis. Vascular smooth muscle cells (VSMC) were harvested from LZ and ZDF rats and exposed to NO. After 24hrs, Western blot analysis for PA28-α and USP5, or the 26S activity assay was performed.

RESULTS:
ZDF arteries exhibited 2-fold more PA28-α expression versus STZ arteries. While NO caused a 2-fold decrease in PA28-α after injury in LZ arteries, NO caused complete inhibition in STZ and ZDF arteries versus injury alone. Similarly, ZDF arteries showed 2-fold more USP5 expression versus STZ arteries. While NO caused a decrease in USP5 expression in LZ arteries, NO exposure resulted in a doublet formation and no inhibition of USP5 in STZ and ZDF arteries versus injury alone. NO inhibited 26S proteasome activity (p<0.05), but there were no differences between LZ and ZDF VSMC. NO had no effect on USP5 expression in LZ and ZDF VSMC. However, treatment of VSMC with NO caused a concentration-dependent decrease in LZ and increase in ZDF PA28-α expression.

CONCLUSION:
NO caused significant early changes in PA28-α and USP5 in rodent models of type 1 and 2 diabetes versus non-diabetic rats. NO also increased expression of PA28-α in diabetic VSMC, but did not differentially affect proteasomal activity or protein deubiquitination in vitro. These data indicate a strong relationship between protein ubiquitination and degradation and the development of neointimal hyperplasia in diabetic rat models and may lead to a novel mechanism to prevent restenosis in diabetic patients.
Abstract 153: Bile Acid Receptor TGR5 Agonism Represents Anti-inflammatory Effects via Stimulating Nitric Oxide Production in Vascular Endothelial Cells
Authors: Kida Taiki , Tsubosaka Yoshiki , Hori Masatoshi , Ozaki Hiroshi , Murata Takahisa
Published: July 2021

Objective
TGR5, a membrane-bound, G-protein-coupled receptor for bile acids, is known to be involved in regulation of energy homeostasis and inflammation. However, little is known about the function of TGR5 in vascular endothelial cells. In the present study, we examined whether TGR5 agonism represents anti-inflammatory effects in vascular endothelial cells focusing on nitric oxide (NO) production.

Methods and Results
In human umbilical vein endothelial cells (HUVECs), treatment with taurolithocholic acid (TLCA), which has the highest affinity to TGR5 among various bile acids, significantly reduced tumor necrosis factor (TNF)-α-induced vascular cell adhesion molecule (VCAM)-1 protein expression and adhesion of human monocytes, U937. These effects were abrogated by a NO synthase (NOS) inhibitor,
N
G
-Monomethyl-L-arginine (L-NMMA). In bovine aortic endothelial cells (BAECs), treatment with TLCA as well as lithocholic acid, which also has high affinity to TGR5, significantly increased the NO production. In contrast, deoxycholic acid and chenodeoxycholic acid, which possess low affinity to TGR5, did not affect the NO production. Gene depletion of TGR5 by siRNA transfection abolished TLCA-induced NO production in BAECs. TLCA-induced NO production was also observed in HUVECs measured as intracellular cGMP accumulation. We next investigated the signal pathways responsible for the TLCA-induced NO production in endothelial cells. Treatment with TLCA increased endothelial NOS (eNOS)
ser1177
phosphorylation in HUVECs. This response was accompanied by increased Akt
ser473
phosphorylation and intracellular Ca
2+
([Ca
2+
]
i
). Treatment with phosphoinositide 3-kinase (PI3K) inhibitor, LY294002, or blockade of calcium channel with La
3+
, significantly decreased TLCA-induced eNOS
ser1177
phosphorylation and subsequent NO production.

Conclusion
These results indicate that TGR5 agonism can mediate anti-inflammatory responses by suppressing VCAM-1 expression and monocytes adhesion to endothelial cells. This function is dependent on NO production via Akt activation and [Ca
2+
]
i
increase.
Abstract 179: Atheroprone Hemodynamics and Oxidized Phospholipids Enhance Endothelial and Smooth Muscle Inflammatory Signaling
Authors: Feaver Ryan E , Ding Kuan-fu , Hastings Nicole , Wamhoff Brian , Blackman Brett R
Published: July 2021

Atherosclerosis is a focal inflammatory disease marked by activation of NFkB within regions where low and oscillating shear stresses (Atheroprone) “prime” the endothelium for an inflammatory response. Interestingly, oxPAPC, a major component of oxLDL, has not been shown to act through the canonical NFkB pathway, and although oxLDL is capable of increasing the expression of NFkB-dependent genes it often requires higher concentrations of oxLDL (>100ug/ml). Further, these studies were all performed within static monocultures, where the endothelium was not a “primed” physiologic phenotype. Therefore, we wanted to test whether the addition of oxLDL within Atheroprone hemodynamics (where NFkB signaling was already activated) would further enhance inflammatory signals downstream of NFkB in an endothelial cell (EC) and smooth muscle cell (SMC) co-culture. After preconditioning cells with Atheroprone shear stress, NFkB activity and gene expression is significantly increased compared to cells exposed to shear stress patterns from regions protected from atherosclerosis. Adding oxLDL to this system further enhances inflammatory gene expression in a dose-dependent fashion above Atheroprone conditions treated with native LDL (nLDL). Performing gene arrays we found that oxLDL regulated over 350 genes in ECs compared to cells treated with nLDL. These gene panels were enriched with shear stress-regulated, pro- (VCAM, E-Selectin) and anti-(KLF2, eNOS) inflammatory genes. Though not directly exposed to either Atheroprone shear stress or LDL, over 50 genes were regulated by these conditions in the underlying SMCs in our co-culture system, including inflammatory genes. Lastly, we measured changes in protein secretion due to the addition of oxLDL compared to nLDL. Many NFkB-dependent, pro-inflammatory cytokine levels were increased, including IL8, MCP-1, and IL6. Investigating the role of oxLDL within the context of physiologic shear stress was found to enhance the Atheroprone-regulated gene profile towards a more pro-inflammatory phenotype. These conditions mimic vessel wall inflammation found in human arteries and provide an ideal environment for testing drugs intended for treating advanced atherosclerosis.
Abstract 183: Oxidized LDL Cholesterol Induces Endothelial Dysfunction in the Aorta and Lox-1 Expression in Macrophages
Authors: Leuner Anja , Poitz David M , Augustin Robert , Neubauer Heike , Brunssen Coy , Ravens Ursula , Strasser Ruth H. , Morawietz Henning
Published: July 2021

Elevated plasma cholesterol is one of the major risk factors in the development of atherosclerotic lesions. Oxidation of native LDL cholesterol (nLDL) by reactive oxygen species leads to the formation of oxidized LDL (oxLDL). An important receptor for the cellular uptake of oxLDL is the lectin-like oxidized low-density lipoprotein receptor-1 (Lox-1). Lox-1 is highly expressed on macrophages, but also present on arterial endothelial and vascular smooth muscle cells. Especially the uptake by macrophages leads to the formation of foam cells in atherosclerotic lesions.

Aim of the present study was to analyze the impact of oxLDL on endothelial function in murine aortas and on Lox-1 expression in human macrophages. In addition, we analyzed the effect of a high-fat diet on vascular function in mice with an endothelial Lox-1 overexpression. First, we incubated aortic rings of wild-type mice for 2 h with 100 μg/mL oxLDL and analyzed the endothelial function using a Mulvany myograph. Compared to basal conditions, oxLDL significantly impaired endothelium-dependent vasodilation. Next, we fed mice with an endothelial overexpression of Lox-1 for 20 weeks a high-fat diet and analyzed the endothelial function in the thoracic aorta. Interestingly, these mice had no impaired endothelium-dependent relaxation after high-fat diet feeding. To get further insight into Lox-1 regulation by oxLDL, we analyzed the impact of oxLDL on Lox-1 expression in human macrophages. Monocytic THP-1 cells were differentiated with phorbol myristate acetate into macrophages and stimulated for 24 h with nLDL or oxLDL. We found a significant induction of Lox-1 mRNA expression after oxLDL incubation, whereas nLDL had no effect. Our data suggest an increased oxLDL uptake in oxLDL-treated macrophages by increased Lox-1 receptor expression.

In conclusion, our data support an important role of oxLDL as a proatherosclerotic risk factor by its ability to induce endothelial dysfunction and Lox-1 expression in macrophages. Both processes may be involved in the development of atherosclerotic lesions but the physiological significance and functional role of Lox-1 in contributing to the human disease warrants further investigations.
Abstract 207: Differential Regulation of Monocyte/Macrophage Atherogenic Properties by Adiponectin: Role of Macrophage Polarization and Adiponectin Receptors
Authors: vanStijn Caroline M , Kim Jason , Tangirala Rajendra K
Published: July 2021

Adiponectin, an adipocytokine produced by the adipose tissue, exerts metabolic, anti-inflammatory and anti-atherogenic effects to ameliorate diabetes and cardiovascular disease and is a potentially important therapeutic target. However, mechanisms of adiponectin vascular actions and the regulation of macrophage adiponectin receptor expression under inflammatory/atherogenic activation remain unclear. Our studies with human monocytes/macrophages revealed differential adiponectin receptor regulation in subjects with insulin-resistance. Here, we investigated adiponectin regulation of macrophage gene expression under pro- and anti-inflammatory conditions. We addressed the hypothesis that differential activation of macrophages into the classical (M1) or alternative (M2) program alters their adiponectin receptor (AdipoR1 and AdipoR2) expression. The microarray gene expression analyses in human monocytes exposed to TNF-α showed that adiponectin inhibited several inflammatory/atherogenic genes. Our studies revealed that adiponectin itself induces AdipoR1 and AdipoR2 expression in macrophages. We further investigated the effects of macrophage polarization (M1 or M2) on adiponectin receptor expression in bone marrow-derived and peritoneal macrophages. These studies demonstrated that M1 activation (IFN-γ and LPS) significantly reduced AdipoR1 and AdipoR2 expression. In contrast, M2 activation of (IL-4 or IL-10) maintains a significantly higher level of AdipoR1 and AdipoR2. In M2 activation, adiponectin receptor expression was more substantial in IL-10 than IL-4-polarized macrophages. These results provide important evidence that macrophage polarization profoundly alters their adiponectin receptor expression and thus functional responses to adiponectin. Thus, adiponectin-mediated macrophage functions are regulated by adiponectin receptor expression which is modulated by the macrophage polarization which controls their inflammatory and atherogenic properties.
Abstract 213: Ticagrelor Inhibits Human Platelet Aggregation via Adenosine in Addition to P2Y12 Antagonism
Authors: Nylander Sven , Femia Eti , Scavone Mariangela , Berntsson Pia , Asztély Anna-Karin , Löfgren Lars , von Bahr Helena , Nilsson Ralf G , Cattaneo Marco
Published: July 2021

Introduction.
Ticagrelor, a P2Y
12
antagonist, inhibits adenosine uptake by erythrocytes and increases adenosine-induced coronary blood flow.

We hypothesiszed that ticagrelor inhibits platelet function not only by antagonizing P2Y
12
, but also by increasing the levels of adenosine, which inhibits platelets by interacting with the platelet A
2A
receptor.

Methods.
Collagen-induced platelet aggregation (PA) was measured in citrate-anticoagulated whole blood (WB) (Multiplate) or platelet-rich plasma (PRP) (light-transmission aggregometry) from 12 healthy controls and 2 patients with congenital deficiency of P2Y
12
, in the presence or absence of adenosine (7.1 μM), which marginally affected PA in WB, and the A
2A
inhibitor ZM241385. The effects on PA of ticagrelor, the active metabolite of prasugrel (PAM) (P2Y
12
antagonist) and dipyridamole (DYP) (ENT-1 inhibitor) were compared. The effects of these drugs on the clearance of adenosine after its in vitro addition to WB were also studied.

Results.
PA in WB: Adenosine contributed to drug-induced inhibition of PA in healthy controls. The magnitude of the contribution (absolute difference in % aggregation) was greater in the presence of ticagrelor (20%, p <0.01) or DYP (18%, p <0.01) than in the presence of PAM (6%, p 0.02). PA in PRP: Adenosine markedly contributed to inhibition of PA in healthy controls in the presence of all tested drugs. In both WB and PRP ZM241385 reversed the inhibitory effect of adenosine. Data from P2Y
12
-deficient patients was in agreement, but due to low numbers no statistical analysis was performed. Adenosine, added at 7.1μM to WB in vitro, was detectable up to 0.5 min in the presence of vehicle or PAM, up to 3 min in the presence of ticagrelor, and up to 60 min in the presence of DYP. These results are compatible with the hypothesis that ticagrelor and DYP, but not PAM, increase the adenosine levels by inhibiting its uptake by erythrocytes.

Conclusion.
This study provides the first evidence of an additional anti-platelet mechanism by ticagrelor, mediated by the induced increase of adenosine levels in WB. It remains to be established if and to what extent this additional mechanism of action contributes to the strong clinical benefit observed with ticagrelor in patients with acute coronary syndromes.
Abstract 266: MicroRNA Profiling Identifies Enrichment for Proteoglycan-Genes in Symptomatic Carotid Plaques
Authors: Adhikari Neeta , Carlson Marjorie , Lerman Amir , Loeffler Darrell , Walek Dinesha , Daniel Jerry , Mackey Aaron J , Lawson Mark J , Hall Jennifer L
Published: July 2021

MicroRNAs are a class of small non-coding RNAs that regulate gene expression by binding to discrete sites of genes and inhibiting translation. The objective of this study was to identify microRNAs and downstream enriched targets associated with symptomatic carotid plaques. Carotid plaque samples from patients were defined as symptomatic if they were associated with a prior ischemic stroke or transient ischemic attack, or as asymptomatic if there were no prior ischemic events. Digital microRNA profiling was performed on the NanoString nCounter analysis system (NanoString Technologies, Seattle, WA) on carotid plaques from symptomatic (n=6) and asymptomatic (n=6) patients. We used the BioConductor/R package "edgeR" to perform moderated-ANOVA testing for differences in gene abundance between groups, assuming a negative binomial distribution of gene counts, with Benjamini-Hochberg multiple hypothesis correction to control false discovery rate. Analysis revealed five microRNAs that were significantly up-regulated in the symptomatic cohort (see Table 1) including miR-16, which was confirmed by RTqPCR (p=0.02). Predicted downstream targets of miR-16 were enriched for proteoglycan related genes including hyaluronan synthase 2, perlecan and betaglycan. Additionally, RTqPCR demonstrated up-regulation of CathepsinB transcript in symptomatic patients (n=46-50 per cohort, p=0.02) while CathepsinL and heparanase transcripts were not significantly different. CathepsinL enzyme activity was also not significantly different (n=46-50 per cohort). Taken together, these findings may reveal novel proteoglycan mediated signaling pathways, mediated through miR-16, regulating the progression of carotid plaque formation.
Abstract 305: HDL Antagonizes the Defects in the Lymphatic System that Contribute to the Pathogenesis of Rheumatoid Arthritis
Authors: Bisoendial Radjesh , Weninger Wolfgang , Petrides Francine , Tabet Fatiha , Tak Paul P , Rye Kerry Anne
Published: July 2021

Rheumatoid arthritis (RA) is a common inflammatory erosive arthropathy that increases cardiovascular (CV) risk two-fold. Lymphatic dysfunction, a hallmark of chronic inflammation, is considered an attractive target to resolve synovial inflammation. Given the strong vascular anti-inflammatory properties of high-density lipoprotein (HDL) and its key apolipoprotein (apo)A-I, we hypothesized that they may promote inflammation resolution in RA by improving lymphatic endothelial function via up-regulation of the homeobox-containing transcription factor, Prox-1,a key regulator of inflammation-induced lymphangiogenesis (IIL) .

Primary human dermal lymphatic endothelial cells (LEC) were treated for 3 days with TNFα (10 ng/mL) or pre-incubated with apoA-I (1.2 mg/mL) for 24 h, followed by TNFα exposure for 3 days. Prox1 mRNA levels were quantified by qPCR. For the tube formation assay, LECs were seeded into a pre-coated Matrigel 24-well plate, and treated with TNFα for 6 h or pre-incubated with apoA-I for 24 h, followed by TNFα exposure for 6 h. Mouse thoracic ducts (TD) were isolated from 2-4 month old C57Bl/6 mice for studying lymphatic vessel (LV) sprouting. TD rings (1 mm) were implanted into a 3-D culture system containing Matrigel under hypoxic conditions and either treated with TNFα, or pre-incubated for 24 h with apoA-I then exposed to TNFα.

Incubation with TNFα decreased LEC Prox1 mRNA levels by about 56% (P<0.05). Prior exposure of LECs to apoA-I blocked TNFα-mediated Prox-1 suppression (P<0.05). TNFα also reduced LEC tube formation by about 55% (p<0.05) and completely inhibited LV sprouting from TD rings (p<0.05). ApoA-I protected against TNFα-mediated inhibition of LEC tube formation (44.7±8.1 versus 81.9±15.0% of control; P<0.05) and the inhibitory effect of TNFα on LV sprouting (P<0.05).

These results suggest that apoA-I directly regulates IIL by upregulating Prox1 and protecting against TNFα-mediated restriction of lymphatic sprouting and growth in vitro. Hence, raising HDL may provide dual therapeutic benefits in RA by targeting inflammation and reducing cardiovascular risk. New classes of HDL-raising drugs will allow us to study the long term effects of increasing HDL levels on RA progression and CV risk.
Abstract 310: Improvement in Vascular Reactivity by Institution of a "Green-Based" Diet with Supplemental Fish Oil and the Polyphenolic Compounds, Grape Seed Extract and Pycnogenol
Authors: Gundry Steven R , Epstein Jean
Published: July 2021

Objectives:
Endothelial dysfunction (ED) is highly correlated with the development of coronary artery and vascular disease in general. While dietary factors have been previously implicated in the causation of endothelial dysfunction, a purposeful change in dietary habits, coupled with the addition of several polyphenol-containing supplements, has not been studied for their effects on endothelial function, as measured by Peripheral Arterial Tonometry (PAT).

Methods:
125 consecutive pts, aged 65-86, M:F ratio 3/2, with known vascular risk factors of HTN, DM, Hypercholesterolemia, hx of MI, Stent, CABG, were enrolled in a dietary program, which emphasizes large amts of leafy green vegetables, olive oil, radical reduction of grain, legumes, and fruits; and generous amts of animal proteins (Diet Evolution). All pts were instructed to take 2-4,000 mg of high DHA fish oil, 200mg of Grape Seed Extract, and 50 mg of Pycnogenol per day. All pts had Endothelial Reactivity (ER) using PAT before and after a 5-minute arm occlusion using the EndoPAT 2000 (Itamar, Israel) at baseline and at 6 months.

Results:
Baseline Endothelial Reactivity was 1.88+/-0.7 (range 1.0-3.3), with 90/125 pts (72%)having endothelial dysfunction (less than 1.60). At 6 months, ER increased to 2.25+/-0.5 (range 1.2-3.6) (p<0.01). Only 20/125 (16%) remained with ED, but all had increased ER numbers. Two pts stopped the polyphenols after a normal PAT; both developed ED on repeat PAT.

Conclusions:
We conclude that a limited grain, legume, fruit diet with emphasis on greens, olive oil and animal proteins with supplemental fish oil and polyphenols dramatically improves endothelial function in a group of high risk patients.
Abstract 316: Deficiency in the Low-Density Lipoprotein Receptor-Related Protein 8 Alters Macrophage Function and Promotes Atherosclerotic Plaque Necrosis
Authors: Waltmann Meaghan D , Basford Joshua E , Konaniah Eddy S , Cash James G , Hui David Y
Published: July 2021

The low-density lipoprotein receptor-related protein 8 (Lrp8; apolipoprotein E receptor 2; ApoER2) is a member of the low-density lipoprotein receptor (
Ldlr
) gene family which is predominately expressed in the brain, but is also expressed in vascular cells and macrophages where its function is currently unknown. Genomewide linkage analysis has associated a single nucleotide polymorphism in the
LRP8
gene with familial and premature coronary artery disease and myocardial infarction in humans, suggesting that Lrp8 may contribute to atherosclerosis. This study assessed the role of Lrp8 in macrophage function and atherosclerosis. Following 24 weeks on a high-fat, high-cholesterol Western-type diet, lesion sizes were similar between
Lrp8
knockout mice on an
Ldlr
deficient background (
Lrp8
-/-
Ldlr
-/-
) and
Ldlr
single knockout (
Ldlr
-/-
) mice, but more complex lesions with extensive necrosis were observed in the

Lrp8
-/-
Ldlr
-/-

mice. The differences in lesion composition were not due to differences in plasma cholesterol levels, but increased expression of the macrophage marker CD68 in the
Lrp8
-/-
Ldlr
-/-
lesions suggested that these differences were due to alterations in macrophage function. When compared to wild-type (WT) cells, mRNA levels of the nuclear receptor PPARγ and PPARγ-responsive genes such as
Cd36, Abca1
, and
Lrp1
were significantly increased in
Lrp8
deficient macrophages. Additionally, the expression of inducible nitric oxide synthase (iNOS) and
Ldlr
were reduced in
Lrp8
deficient macrophages compared to WT cells. Furthermore, following 24 hours in culture there was a reduced number of adherent macrophages found in the
Lrp8
deficient population compared to the WT cells. Consistent with this, phosphorylation of Akt was reduced while phosphorylation of p53 was increased in
Lrp8
deficient macrophages, suggesting that Lrp8 may affect macrophage viability and proliferation via modulation of cell signaling events. Taken together, these data suggest that Lrp8 plays a critical role in macrophage function via regulation of foam cell formation as well as cell proliferation and viability, with reduced expression of Lrp8 significantly contributing to the pathogenesis of atherosclerosis and lesion necrosis.
Abstract 343: Induction of Apoptosis in Human Macrophages by 7-ketocholesterol and Iron Ascorbate
Authors: Leonard Deborah A , Scofield Ashley M , Greene Robert S
Published: July 2021

Inflammation is the new paradigm for the development and progression of atherosclerotic lesions in coronary artery disease. Factors that increase oxidative stress and apoptosis of cells within the lesion contribute to sustaining the inflammatory response. It was hypothesized that redox-active iron and oxysterols, which have both been found in association with atherosclerotic plaques, contribute to oxidative stress and apoptosis by distinct, but overlapping signaling pathways. The human acute monocytic leukemia cell line THP-1 was used as a model system to study the roles of redox-active iron and 7-ketocholesterol in macrophage apoptosis. The ability of both reagents to induce apoptosis in THP-1 macrophages was demonstrated using flow cytometry. The kinetics of poly(ADP-ribose) polymerase (PARP) cleavage as measured by ELISA were also similar. qRT-PCR was used to measure mRNA levels of several pro- and anti-apoptotic genes. While 7-ketocholesterol increased the level of CHOP mRNA (a transcription factor in endoplasmic reticulum (ER)-initiated apoptosis), there was no similar increase in response to iron ascorbate treatment. Expression of other markers of ER stress, like the transcription factors ATF3 and CEBPbeta, was increased by both compounds, but the timing of changes in gene expression was different. TNFalpha regulates both apoptosis and inflammatory cytokine production. Macrophages are major producers of TNFalpha as well as being highly responsive to the cytokine. A comparison of the changes in TNFalpha mRNA and protein levels in response to iron ascorbate and 7-ketocholesterol suggested that while ERK1/2 and NFkB signaling are important in TNFalpha expression, differentially regulated post-transcriptional processes determine the release of TNFalpha by macrophages. These preliminary results suggest that the apoptotic pathways activated by oxysterols and redox-active iron may be different and that these compounds may have additive or synergistic effects on lesion progression.
Abstract 404: Cilostazol Attenuated Angiotensin II-Induced Abdominal Aortic Aneurysms in Apolipoprotein E--Deficient Mice
Authors: Umebayashi Ryoko , Uchida Haruhito A , Makino Hirofumi
Published: July 2021

Objective:
Chronic infusion of angiotensin II (AngII) promotes development of abdominal aortic aneurysms (AAAs) in hypercholesterolemic mice. Previous studies have shown that local inflammation and increased oxidative stress plays an important role on formation of AAAs. Cilostazol, a phosphodiesterase-3 inhibitor with antiplatelet aggregation and vasodilatory effects, exerts vasoprotective effect by inhibiting inflammation and superoxide generation in several experimental studies. The purpose of this study was to evaluate whether cilostazol influenced AngII-induced AAAs.

Methods and Results:
Male apolipoprotein E deficient mice (8-12 weeks old) were fed with either normal diet or cilostazol-containing (0.1% wt/v) diet. After 1 week of cilostazol administration, mice were infused subcutaneously with either AngII (1,000 ng/kg/min, n = 16 -18) or saline (n = 5 - 6) by osmotic minipumps for 4 weeks. AngII equivalently increased systolic blood pressure, irrespective of cilostazol adminstration. Cilostazol had no effect on serum cholesterol concentrations, triglycerides, high-density lipoprotein-cholesterol, body weights, heart rates, and systolic blood pressures. Cilostazol did not affect ex vivo measurement of maximal diameter of abdominal aorta (0.88 ± 0.04 mm vs 0.88 ± 0.03 mm, n.s.) in saline infused-mice. AngII infusion significantly increased ex vivo maximal diameters of abdominal aortas but cilostazol administration attenuated (1.94 ± 0.16 mm vs 1.53 ± 0.17 mm, P < 0.05). In addition, gelatin zymography demonstrated that cilostazol diminished AngII-induced increase in aortic MMP-2 activity (P < 0.05).

Conclusion:
Cilostazol attenuated AngII-induced AAAs in male apolipoprotein E deficient mice.
Abstract 408: Essential Domain Transition Involving Central Helical Repeats Prevent Lipid-Free ApoA-I from Acquiring Lipid
Authors: Pollard Ricquita , Fulp Brian , Thomas Michael , Sorci-Thomas Mary
Published: July 2021

Plasma high density lipoprotein (HDL) concentration is negatively correlated with the occurrence of coronary heart disease in the human population. Because apoA-I is the main protein constituent of HDL, a thorough understanding of apoA-I structural topology is essential for elucidating its ability to package and mobilize cholesterol for catabolism. To determine which of the 10 helical repeats within apoA-I participates in the structural transitions that drive unfolding of the 4-helix bundle, we created several loss of function mutations. Published three-dimensional coordinates of full-length lipid-free apoA-I were used to predict amino acids having spatial separations of 3-5Å within the 4-helix bundle. Based on these predictions, we proposed that specific targeted double cysteine residue substitutions could form disulfide linkages and prevent “opening” of a critical domain required for unfolding of the apoA-I 4-helix bundle when exposed to lipid. To test the importance of helical repeats 4, 5, 6 and 7, double cysteine mutants D103C-R177C apoA-I and F104C-H162C apoA-I were created, expressed and purified using established procedures. Mass spectrometry combined with MS/MS sequencing was used to verify the “locked” disulfide form of each double cysteine substitution mutants. Using 20% SDS-PAGE we show that electrophoretic mobility-shift distinguishes between “locked” or -DTT versus “unlocked” or +DTT form for each of the mutant apoA-I proteins. Particle formation was tested for each mutant by measuring the formation of recombinant HDL (rHDL) using cholate dialysis, as well as, the formation of nascent HDL (nHDL) from ABCA1 expressing cells. Examination of the size of rHDL and nHDL particles formed suggests that unfolding of lipid-free apoA-I to acquire lipid involves the “locked” or restricted helical repeats 4-7. In conclusion, when both double cysteine apoA-I mutants exist in their “locked” conformation evidence of impaired particle formation was observed confirming the existence of the apoA-I 4 helix bundle in lipid free state and the role of central helical repeats 4-7 in lipid binding and particle formation.