bluuebunny/crossref_metadata_2025 huggingface dataset:
Diet and supplement assessment in a Brazilian urban population
Authors: Sousa Alessandra Gaspar , Costa Teresa Helena Macedo da
Published: July 2021

OBJECTIVE: To assess total usual nutrient intakes from foods and dietary supplements by age, sex, physical activity, and nutritional status, and to compare usual nutrient intakes to the Dietary Reference Intakes among non-users and users of dietary supplements in an urban population. METHODS: Cross-sectional population-based survey with 506 adults conducted in the city of Brasília, Brazil, using 24h food recalls. The 24-HR was collected on two nonconsecutive days, for which individuals reported all food, supplements, and beverages consumed in the previous 24 hours. The estimates of mean and the distribution percentiles were adjusted to reflect usual nutrient intake using the Iowa State University method. The prevalence of inadequate micronutrient intake was estimated according to sex using the Estimated Average Requirement (EAR), and values above the Tolerable Upper Intake Level (UL) were also considered. Also, a comparison was made of the total mean usual intake between supplement users and non-users according to BMI and physical activity. RESULTS: The total mean usual dietary intake was significantly higher among users than non-users of dietary supplements (p ≤ 0.02). Dietary supplement use increased intakes of nutrients and decreased prevalence of inadequacy according to sex, with only small (typically < 13%) increases in the population exceeding the Tolerable Upper Intake Level. There was a significant interaction between physical activity and BMI categories with supplement use. CONCLUSIONS: The population that consumes food supplements comprises individuals with more advanced age, female, normal BMI, and physically active. Our findings show that the use of supplements appears beneficial to attain nutrient adequacy. Careful monitoring of intake from food and supplements is recommended, and the statistical methods must be powerful enough to achieve relevant information.
Satisfação dos usuários em relação aos serviços de Atenção Primária à Saúde no Brasil
Authors: Cantalino Juliana Leal Ribeiro , Scherer Magda Duarte dos Anjos , Soratto Jacks , Schäfer Antônio Augusto , Anjos Davllyn Santos Oliveira dos
Published: July 2021

OBJETIVO: Analisar a satisfação de usuários em relação ao acesso, à infraestrutura e à qualidade dos serviços na Atenção Primária à Saúde (APS) no Brasil. MÉTODOS: Estudo transversal com dados de 114.615 usuários vinculados a 30.523 equipes de saúde, obtidos por meio do banco de dados do Programa Nacional de Melhoria do Acesso e da Qualidade da Atenção Básica (PMAQ-AB). Foram estudadas variáveis independentes relacionadas ao acesso, à infraestrutura e à qualidade dos serviços na APS. O desfecho, satisfação dos usuários, foi mensurado através das variáveis: “se tivesse a opção, mudaria de equipe ou de serviço de saúde” e “recomendaria este serviço de saúde para um amigo ou familiar”. Para avaliar a satisfação de acordo com as variáveis independentes de exposição, foi utilizado o teste Qui-quadrado de Pearson, considerando nível de significância de 5%. Análises descritivas das variáveis foram realizadas através de frequências absoluta (n) e relativa (%). RESULTADOS: A satisfação dos usuários esteve associada às variáveis de acesso (p < 0,001), infraestrutura (p < 0,001) e qualidade dos serviços (p < 0,001) na APS. A proximidade do serviço, atenção à demanda espontânea, escuta e o respeito dos profissionais às singularidades do paciente, assim como a resolutividade dos serviços, sem necessidade de encaminhamentos para outros e a boa infraestrutura, estiveram relacionados com a satisfação dos usuários. CONCLUSÃO: Para a garantia da melhoria da qualidade dos serviços ofertados na APS do Brasil, os aspectos de satisfação dos usuários identificados no presente estudo devem ser considerados na organização e gestão dos serviços.
Análise da estrutura fatorial do Audit em adolescentes entre 18 e 19 anos
Authors: Machado Patrícia Maria Abreu , Campelo Cleber Lopes , Oliveira João Victor Pimentel de , Batista Rosângela Fernandes Lucena , Simões Vanda Maria Ferreira , Santos Alcione Miranda dos
Published: July 2021

OBJETIVO: Determinar a estrutura fatorial do instrumento Alcohol Use Disorders Identification Test (Audit) em uma amostra representativa de adolescentes de 18 a 19 anos. MÉTODOS: Estudo transversal realizado com adolescentes nascidos em São Luís (MA). A consistência interna do instrumento foi determinada pelo coeficiente alfa de Cronbach e a validade do construto foi avaliada por meio da Análise Fatorial Confirmatória (AFC). O índice de Kaiser-Meyer-Olkin (KMO) foi calculado para analisar a adequação da amostra. A qualidade de ajuste do modelo fatorial foi analisada de acordo com os índices dos testes qui-quadrado de ajustamento, comparative fit index (CFI), Tucker-Lewis index (TLI) e root mean square error of approximation (RMSEA). RESULTADOS: A amostra do estudo foi composta por 1.002 adolescentes entre 18 e 19 anos, sendo 56,8% meninas, 68,5% com 18 anos, 63,3% pardos ou mulatos, 48,6% pertencentes à classe C, 15,4% não trabalhavam e não estudavam e 52,1% tinham pais separados. A amostra foi adequada para a análise fatorial confirmatória (KMO = 0,79) e o coeficiente do alfa de Cronbach foi de 0,70, demostrando consistência interna satisfatória com cargas fatoriais acima de 0,5, com exceção do item 9 “Ficou ferido ou ficou alguém ferido por ter bebido”. A análise fatorial confirmatória revelou a validade do modelo de três fatores para a amostra em estudo com base nos índices de ajustes psicométricos. CONCLUSÃO: A estrutura fatorial do Audit com três fatores foi confirmada para a população de adolescentes entre 18 e 19 anos residentes em São Luís, ratificando os domínios conceituais originais propostos pela Organização Mundial da Saúde. O Audit apresentou-se como um instrumento confiável para a identificação do consumo de álcool.
Influence of center-based child care on development of two-year-olds in a Brazilian cohort
Authors: Leão Otávio Amaral de Andrade , Mielke Gregore Iven , Silveira Mariângela Freitas da , Domingues Marlos Rodrigues , Murray Joseph , Neumann Nelson Arns , Dalabona Caroline Caus , Bertoldi Andréa Dâmaso
Published: July 2021

OBJECTIVE To investigate the association between child care attendance since birth and development in two-years-old Brazilian children. METHODS The study used longitudinal data from the 2015 Pelotas Birth Cohort. The childhood development (cognitive, fine and gross motor skills, and language) at two-years-old children was assessed using INTER-NDA (INTERGROWTH-21st Neurodevelopment Assessment). The child care attendance was measured at ages one and two years and categorized as: a) never attended child care; b) attended some child care (one or two years); and c) always attended child care (one and two years). Demographic, socioeconomic, health, and child stimulation variables were considered as confounders. Crude and adjusted analyses of child care attendance and development were carried out using linear regression. RESULTS Out of the 3,870 infants included in the analyses, around 1/3 attended center-based child care. In crude analyses, attending center-based child care was associated with positive developmental outcomes, except in motor domains. In adjusted analyses, compared to those children that have never attended child care, children who did attend presented higher scores for cognitive development (always in child care: β: 2.44, 95%CI: 0.83–4.05; some child care: β: 1.35, 95%CI: 0.17–2.53). CONCLUSIONS This study suggests that center-based child care may help improve child cognitive development in the Brazilian context. Furthermore, the association was higher for early and continued attendance. Considering the low prevalence of children in external care, it is recommended to improve child care opportunities in early childhood.
Surveillance of hemorrhagic fever and/or neuroinvasive disease: challenges of diagnosis
Authors: Araújo Leonardo José Tadeu de , Gonzalez Lorenzo Lang , Buss Lewis Fletcher , Guerra Juliana Mariotti , Gomez David Salas , Ferreira Camila Santos da Silva , Cirqueira Cinthya Santos , Ghillardi Fábio , Witkin Steven S. , Sabino Ester Cerdeira
Published: July 2021

OBJECTIVE To evaluate the performance of post mortem laboratory analysis in identifying the causes of hemorrhagic fever and/or neuroinvasive disease in deaths by arbovirus infection. METHODS Retrospective cross-sectional study based on the differential analysis and final outcome obtained in patients whose samples underwent laboratory testing for arboviruses at the Pathology Center of the Adolfo Lutz Institute, in São Paulo, Brazil. RESULTS Of the 1355 adults clinically diagnosed with hemorrhagic fever and/or neuroinvasive disease, the most commonly attributed cause of death and the most common final outcome was dengue fever. Almost half of the samples tested negative on all laboratory tests conducted. CONCLUSION The failure to identify the causative agent in a great number of cases highlights a gap in the diagnosis of deaths of unknown etiology. Additional immunohistochemical and molecular assessments need to be added to the post-mortem protocol if all laboratory evaluations performed fail to identify a causative agent. While part of our findings may be due to technical issues related to sample fixation, better information availability when making the initial diagnosis is crucial. Including molecular approaches might lead to a significant advancement in diagnostic accuracy.
COVID-19 and mental health of pregnant women in Ceará, Brazil
Authors: Machado Márcia Maria Tavares , Rocha Hermano Alexandre Lima , Castro Marcia C. , Sampaio Edgar Gomes Marques , Oliveira Francisco Ariclene , Silva Jordan Prazeres Freitas da , Aquino Camila Machado de , Sousa Liduina de Albuquerque Rocha de , Carvalho Francisco Herlanio Costa , Altafim Elisa Rachel Pisani , Correia Luciano Lima
Published: July 2021

OBJECTIVE To assess the perceptions of pregnant women about COVID-19 and the prevalence of common mental disorders during the implemented social distancing period. METHODS This was an observational, cross-sectional study using digital media, of pregnant women exposed to social distancing due to the COVID-19 pandemic, in Fortaleza, Ceará, Northeastern Brazil. Common mental disorders were estimated using the modified Self-Report Questionnaire-20 (SRQ-20) scale, and the feelings towards COVID-19 were assessed using the Fear of COVID-19 scale through telephone calls made in May 2020. COX multivariate regression models were used to verify the associations. RESULTS Of the 1,041 pregnant women, 45.7% (95%CI: 42.7–48.8) had common mental disorders (CMD). All items of the Fear of COVID-19 Scale showed a significant association with the prevalence of CMD (p < 0.001). A CMD risk gradient was observed, going from a prevalence ratio of 1.52 (95%CI: 1.13–2.04) in pregnant women with two positive items to 2.70 (95%CI: 2.08–3.51) for those with four positive items. Early gestational age and the lack of prenatal care were also associated with CMD. CONCLUSIONS The prevalence of common mental disorders in pregnant women was high during the period of social distancing and was aggravated by negative feelings towards COVID-19.
Doenças crônicas não transmissíveis e covid-19: resultados do estudo Epicovid-19 Brasil
Authors: Mesenburg Marilia Arndt , Hallal Pedro Curi , Menezes Ana Maria Baptista , Barros Aluísio J D , Horta Bernardo Lessa , Barros Fernando Celso de , Hartwig Fernando Pires , Jacques Nadège , Silveira Mariangela Freitas da
Published: July 2021

OBJETIVO Descrever a prevalência de doenças crônicas e fatores socioeconômicos e demográficos associados, avaliar os padrões de distanciamento social e a prevalência de anticorpos contra SARS-CoV-2 e sintomas de covid-19 em portadores e não portadores de doenças crônicas. MÉTODOS Foram avaliados dados de 77.075 mil indivíduos de 20 a 59 anos de três etapas do inquérito sorológico de abrangência nacional Epicovid-19 Brasil, realizadas entre maio e junho de 2021. A presença de anticorpos contra SARS-CoV-2 foi avaliada por teste rápido. Foram investigadas as prevalências autorreferidas de hipertensão, diabetes, asma, câncer, doença renal crônica e doença cardíaca. A prevalência de uso de máscara, de adesão a medidas de isolamento e de anticorpos foi avaliada separadamente entre portadores e não portadores de doenças crônicas. A prevalência de sintomas foi avaliada entre doentes crônicos e não doentes portadores de anticorpos. RESULTADOS A prevalência do pelo menos uma doença crônica foi de 43%, maior na região Sudeste, entre indivíduos brancos e indígenas, mulheres, menos escolarizados e em menor posição socioeconômica. O uso de máscara ao sair do domicílio não diferiu entre doentes crônicos e não doentes (98%). A proporção de participantes que referiram adesão ao isolamento foi maior entre doentes crônicos (15,9%) que entre não doentes (24,9%). A prevalência de anticorpos contra SARS-CoV-2 foi semelhante entre doentes crônicos e não doentes (2,4% e 2,3%). A prevalência de tosse, dispneia, palpitações e mialgia foi significativamente maior entre doentes crônicos, mas a proporção de sintomáticos não diferiu entre os grupos. CONCLUSÃO A prevalência de doenças crônicas no país é alta e a pandemia de covid-19 atinge de forma semelhante doentes e não doentes. Doentes crônicos apresentam formas mais graves de covid-19 e maior prevalência de sintomas. A maior adesão às medidas de distanciamento social entre doentes crônicos não se reflete em menor incidência de covid-19 nesse grupo.
Decade of Aortic Valve Sparing Reimplantation
Authors: Kallenbach Klaus , Karck Matthias , Pak Dorota , Salcher Rolf , Khaladj Nawid , Leyh Rainer , Hagl Christian , Haverich Axel
Published: July 2021

Background—

This single center study assesses the outcome of aortic valve sparing reimplantation (AVS) in 284 consecutive patients who were operated on for various indications during the last 11 years.

Methods and Results—

From July, 1993, to July, 2004, 284 patients underwent AVS. Mean age was 53±16 (range 8 to 84) years. Of the 284 patients, 184 were male (64.8%) and 54 (19%) experienced Marfan’s syndrome. Acute aortic dissection Stanford type A was present in 53 patients (19%) and a bicuspid aortic valve was present in 17 patients (6%). Concomitant arch replacement was necessary in 120 patients (42%). Additional procedures were performed in 66 patients (23.2%). Mean follow-up time was 41±32 (range 0 to 130) months. The 30-day mortality was 3.2% overall, 11.3% in emergency patients, and 1.3% in elective patients. Mean bypass time was 174±48 (range 90 to 440) minutes and aortic cross clamp time was 132±33 (range 64 to 283) minutes. In patients undergoing arch replacement, circulatory arrest was 25±17 (range 7 to 99) minutes. Rethoracotomy for bleeding was required in 4.6% of patients. During follow-up, there were 20 (7.3%) late deaths. Reoperation of the reconstructed valve was required in 15 patients (5.3%); underlying reasons were endocarditis (n=4) and aortic insufficiency (n=11) requiring aortic valve replacement. Average grade of aortic insufficiency increased significantly from 0.23±0.46 postoperatively to 0.34±0.59 at latest evaluation (
P
=0.026). Two patients experienced a transient ischemic attack early postoperatively; no further thromboembolic complications were noticed. The majority of patients (96%) presented with a favorable exercise tolerance at last contact.

Conclusions—

The aortic valve reimplantation technique leads to excellent clinical outcome in patients with various pathologies. Lack of anticoagulation and favorable durability should encourage the extension of indications for this technique.
Abstract 119: Potential Role for Bone Marrow Niche and Endothelial Progenitor Cell Dysfunction in Critical Limb Ischemia
Authors: Teraa Martin , Sprengers Ralf W , Moll Frans L , Verhaar Marianne C ,   JUVENTAS Study Group
Published: July 2021

Background

Critical limb ischemia (CLI) is characterized by obstruction of lower extremity arteries and a largely unexplained impaired ischemic neovascularization response. Bone marrow (BM) derived endothelial progenitor cells (EPC) contribute to postnatal neovascularization. We hypothesize that reduced levels and function of circulating progenitor cells and a dysfunctional BM environment contribute to impaired neovascularization in CLI.

Methods

Levels of primitive (CD34+ and CD133+) progenitors and CD34+KDR+ haemangioblastic EPC were analyzed using flow cytometry in peripheral blood (PB) and BM from 101 CLI patients in the JUVENTAS trial (
NCT00371371
) and healthy controls (n=37 and n=12 for PB and BM, respectively). Endothelial damage markers (sE-selectin, sICAM-1, sVCAM-1, thrombomodulin) and PB levels of progenitor cell mobilizing (VEGF, SDF-1α, SCF, G-CSF) and inflammatory (IL-6, IL-8, IP-10) factors were assessed by ELISA and multiplex. Levels and activity of the EPC mobilizing protease MMP-9 were assessed in BM plasma by ELISA and zymography. Circulating angiogenic cells (CAC) were cultured from PB, and CAC paracrine function was assessed.

Results

Endothelial damage markers were higher in CLI (
p<
0.01). PB levels of VEGF, SDF-1α, SCF, G-CSF (
p<
0.05) and of IL-6, IL-8 and IP-10 were higher in CLI (
p<
0.05). Circulating EPC and CD133+ cells and BM CD34+ cells were significantly lower in CLI (all
p
<0.05), BM levels and activity of MMP-9 were lower in CLI (both
p<
0.01). Multivariate regression analysis showed an inverse association between IL-6 levels and BM CD34+ cell levels (
p=
0.007). CAC outgrowth did not differ significantly between CLI patients and healthy controls (
p=
0.137), however CAC from CLI patients had profoundly reduced migration stimulating potential (
p<
0.0001).

Conclusion

CLI patients have reduced levels of circulating EPC despite profound endothelial injury and an EPC mobilizing response. Moreover, CLI patients have lower BM CD34+ cell levels, which were inversely associated with the inflammatory marker IL-6, and lower BM MMP-9 levels and activity. Our data suggest that reduced levels and function of circulating progenitor cells and BM dysfunction contribute to the defective neovascularization response in CLI.
Abstract 145: Epigenetic Modifications of Bone-Marrow Progenitor Cells Promote an M1 Phenotype in Peripheral Macrophages in Diabetes
Authors: Gallagher Katherine A , Joshi Amrita , Carson William , Schaller Matthew , Henke Peter , Feldman Eva , Kunkel Steven
Published: July 2021

Introduction:
Diabetic wounds are characterized by a chronic inflammatory state that is maintained by overexpression of pro-inflammatory cytokines generated by immune cells, namely macrophages. To understand this imbalance, it is critical of understand how changes in immune cells at the systemic level influence the peripheral immune cells. Work from our lab has shown that cytokines induce a histone post-translational “epigenetic signature” at promoter sites of specific genes. However, the role of histone modification of DNA on diabetic progenitor and differentiated immune cells and their impact on chronic inflammation in the periphery has not been studied. We hypothesized that epigenetic changes to bone marrow cells influences peripheral macrophage phenotypes.

Methods:
Bone marrow was harvested from 7 diet-induced obese mice (DIO) (mean glucose= 290) and 7 wildtype (WT) controls. In some of the mice, lin -/ckit+ population was isolated and flow cytometry (FACS) analysis was performed to isolate hematopoietic stem cells (HSC) and myeloid progenitor cells (MYP). HSC were defined as cells expressing sca1+/CD150+/flt3+ and MYP co-expression of sca1+/CD150+/FcgR-/CD105low. HSC and MYP were analyzed via FACS for the presence of chromatin marks. In other mice, BMD macrophages were isolated and propagated in cell culture. Macrophages were either untreated or skewed towards an M1 or M2 phenotype with IF-gamma or IL-4, respectively. Levels of protein and mRNA of M1/M2 products were analyzed. Additionally, at 6, 24 and 72 hours the macrophages were assessed for expression of chromatin remodeling marks via FACS analysis.

Results:
HSC had a 2 fold decrease in the expression of H3K9 histone methylation marks in the DIO mice as compared to WT. (Figure 1A) This pattern was also observed in the more differentiated MYP cells. (Fig 1B) Macrophages isolated from diabetic mice also reflected these histone changes, supporting the role of histone methylation on fully-differentiated macrophage gene expression. (Fig 1C) DIO macrophages demonstrated increased levels of TNF-alpha and CCL3, classic M1 phenotype markers. (Fig 1D)

Conclusions:
Epigenetic changes initiated in the bone marrow progenitor cells effect immune cells in the periphery, and may contribute to the chronic inflammation seen in diabetics. Manipulation of these histone modifications at the bone marrow level could allow for the development of therapies to prevent chronic inflammation in the periphery of diabetics.
Abstract 166: Small Dense LDL Cholesterol Is Associated with Risk for Coronary Heart Disease: The Atherosclerosis Risk in Communities (ARIC) Study
Authors: Gaubatz John W , Sun Wensheng , Jiang Jennifer , Buchanan Ashley , Couper David , Virani Salim , Boerwinkle Eric , Ballantyne Christie M , Hoogeveen Ron C
Published: July 2021

Background—
Evidence from
in vitro
studies indicates that small dense LDL (sd-LDL) is more atherogenic than large buoyant LDL. Previously, sd-LDL has been associated with risk for vascular disease. However, the lack of a standardized sd-LDL assay has hampered its clinical application.

Objectives—
We tested the hypothesis that elevated plasma sd-LDL-cholesterol (sd-LDL-C) level is associated with risk for incident coronary heart disease (CHD) and stroke in the ARIC cohort.

Methods—
Plasma sd-LDL-C was measured in 11,419 men and women of the biracial ARIC study using a newly developed automated homogeneous assay. A proportional hazards model was used to examine the relationship between sd-LDL-C, vascular risk factors, and risk for CHD events and stroke over a period of ≈10 years.

Results—
Mean plasma sd-LDL-C was higher in Caucasians than in African Americans (45.2 vs. 37.4 mg/dL, p<0.0001). Plasma sd-LDL-C levels were strongly correlated with an atherogenic lipid profile and were higher in diabetics vs. non-diabetics (49.6 vs. 42.3 mg/dL, p<0.0001, respectively). sd-LDL-C was associated with incident CHD in a basic model as well as a model that included traditional risk factors and hs-CRP with hazard ratios (HRs) of 1.99 (95%CI: 1.68-2.36) and 1.56 (95%CI: 1.26-1.93) for the highest vs. the lowest quartile, respectively (Table). We did not find a significant association of sd-LDL-C with risk for stroke (Table).

Conclusions—
sd-LDL-C is associated with incident CHD but does not predict risk for stroke in ARIC participants. Further studies will need to determine whether sd-LDL-C will add value beyond traditional risk factors to cardiovascular risk assessment in clinical practice.
Abstract 168: Effects of the CETP Inhibitor Anacetrapib on HDL3-to-HDL2 Transfer: Comparison of in Vitro and in Vivo Methodologies
Authors: Johns Douglas G , Chen Ying , Daniels Erin D , Castro-Perez Jose , Wang Sheng-Ping , Akinsanya Karen O
Published: July 2021

Cholesteryl ester transfer protein (CETP) is a target for the treatment of dyslipidemia and coronary artery disease. In addition to the well-known effect of CETP to transfer CE from HDL to LDL and to VLDL,
in vitro
, CETP has been reported to transfer CE between small and large HDL particles (HDL2 and HDL3, respectively). We sought to understand how the CETP inhibitor anacetrapib (ANA) affects HDL3-to-HDL2 transfer under both
in vitro
and
in vivo
conditions.
In vitro
, ANA dose-dependently inhibited transfer of
3
H-CE from total HDL to LDL (IC50 30nM), and from isolated HDL3 to HDL2 particles (IC50 200nM). In human CETP transgenic mice, animals treated with a single dose of ANA (100mg/kg) displayed 80% maximal reduction in plasma CETP activity and a 22% increase in total HDL cholesterol. In animals treated with either vehicle or ANA,
3
H-CE-labeled HDL3 was injected intravenously and
3
H-tracer was monitored in lipoprotein fractions following injection. Animals treated with ANA showed an increase in the amount
3
H-tracer present in HDL2 compared to vehicle over time (20-70% increase across 6 hrs post
3
H-CE-HDL3 injection, P<0.05 vs vehicle). The HDL2 CE pool was also increased with ANA treatment, and
3
H-cholesterol flux into HDL2 was increased with ANA treatment when adjusted to the change in pool size (at 2 and 4 hrs post 3H-CE-HDL3 injection). No change in HDL2
3
H-tracer was seen in C57BL6 mice (lacking CETP) treated with ANA. These results indicate that in contrast to
in vitro
findings, ANA increases flux of CE into HDL2
in vivo
, a process which likely involves multiple pathways. Therefore, the
in vitro
phenomena of 1) HDL3-to-HDL2 transfer by CETP and 2) inhibition of this transfer by CETP inhibitors are not recapitulated
in vivo
. It is clear that
in vivo
approaches are necessary to understand the relevance of HDL3-to-HDL2 transfer
in vivo
, and to accurately study the effects of CETP inhibition on lipoprotein metabolism.
Abstract 180: Increased Atherosclerosis Associated with Periodontal Disease Is Mediated by CD36
Authors: Febbraio Maria , Brown Paul M
Published: July 2021

We previously showed that inflammation, and not hyperlipidemia alone, was necessary for CD36 dependent atherogenesis. Chronic periodontal disease is characterized by a persistent inflammatory state and is epidemiologically associated with cardiovascular disease. We hypothesize that CD36 is an essential link between periodontal disease and atherosclerosis.

Low density lipoprotein receptor knock out (LDLR KO) mice and CD36/LDLR double KO mice were infected with the periodontal disease associated bacteria, Porphyromonas gingivalis (Pg), by oral lavage and fed a Western diet for 12 weeks (n = 7-14/group). We assessed periodontal disease, risk factors associated with atherosclerosis, and lesion burden. We conducted studies in isolated macrophages to understand mechanistic differences between the groups.

Wild type and CD36 KO macrophages equally phagocytosed bacteria. We measured the cemento-enamel junction of each molar to assess periodontal disease and found that it was significantly increased in infected mice compared with uninfected controls. Histological analysis showed neutrophil, osteoclast and macrophage infiltrates in the alveolar bone of infected mice. Differences in plasma cholesterol, triacylglycerol, insulin resistance and weight gain did not necessarily track with atherosclerosis burden, however blood neutrophils and cytokines were increased in infected LDLR KO mice compared with all other groups. Infected LDLR KO mice had significantly increased atherosclerotic lesion burden compared with uninfected LDLR KO mice, and all of the increased lesion was CD36 dependent. PCR analysis found no evidence for direct infection of atherosclerotic lesions by Pg. In vitro macrophage studies showed that heat killed Pg, lipopolysaccharide (LPS) derived from Pg, oxidatively modified LDL or plasma from infected mice, could not activate the NALP3 inflammasome. Combining heat killed Pg or Pg LPS with oxidatively modified LDL or plasma from infected mice, however, led to significant IL-1 beta secretion that was CD36 and NFkB dependent. Our data suggest that atherosclerosis associated with periodontal disease is mediated by cellular inflammatory responses involving both CD36 and Toll-like receptor.
Abstract 202: Hydrogen Peroxide Increases Osteopontin Expression Through Activation of Transcriptional and Translational Pathways
Authors: Lyle Alicia N , Remus Ebony W , Fan Aaron E , Taylor W Robert
Published: July 2021

The occlusion of blood vessels in the setting of cardiovascular disease leads to ischemia, initiating processes that promote neovascularization to restore blood flow and preserve tissue function. Our in vivo studies show that Osteopontin (OPN) is a critical mediator of post-ischemic neovascularization and that ischemia-induced increases in OPN expression are H
2
O
2
-dependent. However, the mechanisms by which H
2
O
2
increases OPN expression are poorly defined. To determine if H
2
O
2
mediates transcriptional, post-transcriptional, and/or translational regulation of OPN expression in vitro, we used rat aortic smooth muscle cells as an in vitro system and stimulated with H
2
O
2
. Dose response studies showed OPN expression increased with 50 μM H
2
O
2
(51.9%±2.2, p<0.05). Using 50 μM H
2
O
2
, we performed time courses and measured OPN mRNA by qRT-PCR and protein by Western blot. OPN mRNA levels significantly increased in response to H
2
O
2
at 8 (70.4%±5.7, p<0.05) and 18 hours (120.2%±5.2, p<0.005). Interestingly, the increases in OPN protein expression in response to H
2
O
2
occurred in an unusual bi-phasic pattern, with significant increases at 6 (96.9%±1.5, p<0.001) and 18 hours (234.0%±3.6, p<0.001), with a return to baseline in between. An increase in OPN mRNA preceded the increase in OPN protein at 18 hours, suggesting transcriptional regulation; however, the acute increase in OPN at 6 hours was not preceded by increased mRNA, suggesting multiple mechanisms of OPN regulation by H
2
O
2
. To determine if the increase in OPN at 6 hours is due to increased mRNA stability or translation, we performed an RNA stability assay. H
2
O
2
stimulation did not alter OPN stability or the rate of OPN RNA degradation, leading us to conclude the increase in OPN expression at 6 hours is due to increased translation. Further studies reveal H
2
O
2
-mediated increases in phosphorylation of 4E-BP1 at the redox-sensitive Ser65 site (89.4%±6.1, p<0.05), allowing for the subsequent release of eukaryotic initiation factor eIF4E and increased phosphorylation at Ser209 (139.2%±3.9, p<0.05), resulting in increased OPN translation. In conclusion, H
2
O
2
enhances OPN expression through acute increases in translation, while long-term increases in OPN occur through increased transcriptional regulation.
Abstract 218: Fibrinolytic Reactivity of Venous Endothelial Cells from Different Vascular Beds
Authors: Kumar Neil G , Roztocil Elisa , Cullen John P , Gillespie David L
Published: July 2021

Objective:
Little is known about the molecular biology of endothelial cells from different venous vascular beds. As a result, our treatment of deep vein thrombosis (DVT) and pulmonary artery embolism (PE) remain identical. PAI-1 and tPA are important regulators of thrombosis and fibrinolysis, while ICAM-1 is known to bind fibrinogen. Here, we aim to investigate differences in fibrinolytic reactivity between human iliac vein endothelial cells (HIVECs) and human pulmonary artery endothelial cells (HPAECs).

Methods:
Confluent HIVECs and HPAECs, passages 3 - 6, were cultured in the absence or presence of TNFα (10 ng/mL) for 24 hours. Cellular expression of tPA and PAI-1 as analyzed by Western blot analysis and ICAM-1 as analyzed by flow cytometry were compared to controls.

Results:
Following TNFα stimulation, PAI-1 was upregulated in both HPAECs and HIVECs, however the upregulation observed in HPAECs was approximately 9-fold the increase observed in HIVECs (relative expression: 3.23 ± 0.52 vs 1.26 ± 0.27, n = 3, p < 0.05). While TNFα had no effect on tPA expression in HIVECs, tPA expression in HPAECS was upregulated by 33% (n = 3, p < 0.05). Although TNFα stimulation increased the number of ICAM-1 positive to approximately 100% in both cell types, a 3-fold greater increase in the Mean Fluorescence Intensity (MFI) was observed in HIVECs when compared to HPAECs (relative MFI: 69.28 ± 13.58 vs 21.92 ± 7.22, n = 3, p <0.05).

Conclusions:
HPAECs and HIVECs react differently in terms of fibrinolytic potential when challenged with a cytokine associated with systemic inflammation, such as in DVT and PE. These findings suggest that endothelial cells from distinct venous vascular beds may differentially regulate the fibrinolytic pathway, thus demonstrating unique properties of the deep veins and the pulmonary artery to respond to thromboembolism.
Abstract 22: LDLR Promotes and PCSK9 Inhibits LDL-Derived Transintestinal Cholesterol Excretion
Authors: Le May Cedric , Berger Jean Mathieu , Pillot Bruno , Prieur Xavier , Letessier Eric , Collet Xavier , Lespine Anne , Cariou Bertrand , Costet Philippe
Published: July 2021

Direct transintestinal cholesterol excretion (TICE) is an alternative path to biliary secretion and accounts for 33% of fecal cholesterol excretion in mice.

Objectives:
We aimed at identifying i) the lipoproteins involved in TICE ii) the role of intestinal LDL receptor (LDLR) and of its natural inhibitor, circulating proprotein convertase subtilisin kexine type 9 (PCSK9) at the basolateral pole iii) the implication of ATP binding cassette transporter ABCB1ab, a cholesterol floppase localized at the enterocytes apical side.

Methods:
We labelled human lipoproteins with free 3H cholesterol (free3H) or with 3H-cholesteryl oleate (3HCO). We measured lipoprotein-derived 3H-cholesterol TICE ex vivo in intestinal explants mounted in Ussing chambers. Human explants were obtained from 4 patients undergoing bariatric surgery with their informed consent. In vivo, TICE was measured in mice i.v. injected with radiolabelled lipoproteins, by cannulation of the proximal intestine and concomitant surgical bile diversion, counting radioactivity in intestinal perfusates over 120 minutes.

Results:
For the first time, we showed direct evidence of TICE in human duodenal explants, from both LDL and HDL. Both lipoproteins (labelled with free3H or
3HCO
) contributed to TICE in mouse explants; TICE was highly responsive to changes in temperature and medium oxygenation. LDL-derived TICE was decreased by 58% (p<0.05) in explants from LDLR knockout mice (LDLR KO), compared with control C57Bl6J. In vivo free3H- or 3HCO-LDL-derived-TICE was conserved in LDLRKO mice, suggestive of a compensatory mechanism. LDL-derived TICE was increased by 62% (p<0.01) in PCSK9KO that present with ∼300% more intestinal LDLR. Acute depletion of intestinal LDLR with purified recombinant PCSK9 (i.v.) led to 40% (p<0.05) less TICE in PCSK9KO but had no effect in LDLRKO, confirming the implication of this receptor. Lovastatin (0.02% W/W 10d) increased TICE by 71% (p < 0.05) in C57Bl6J but not in LDLRKO. Interestingly, using 3H-cholesterol diluted in intralipid as a source, we showed that ABCB1 plays an important role in TICE. Indeed ABCB1ab
-/-
mice presented with 26% (p<0.05) less TICE than FVB controls in vivo and ABCB1 inhibitor PSC-833 (5mircoM) decreased TICE by 64% in an ABCB1 dependent fashion in explants.

Collectively, these results provide the first molecular understanding of TICE.
Abstract 232: The P2Y2 Nucleotide Receptor Mediates Blood Flow Recovery Following Hindlimb Ischemia in Mice
Authors: McEnaney Ryan M , Madigan Michael C , Sachdev Ulka , Billiar Timothy R , Tzeng Edith
Published: July 2021

Objectives:
Arteriogenesis is a process by which small resistance vessels mature into large caliber collateral arteries in response to arterial occlusive disease. The forces stimulating arteriogenesis nclude shear stress and circumferential wall stress. Nucleotides are released from cells in response to mechanical stimuli and may activate nucleotide receptors. Thus, we hypothesize that the P2Y
2
nucleotide receptor, expressed by vascular cells, mediates arteriogenesis.
Methods:
C57Bl6 NJ (WT) and P2Y
2
R KO mice (n=8/group) underwent unilateral femoral artery ligation (FAL). Perfusion was measured immediately after ligation and at 3, 7, 14, 21, and 28 days using laser Doppler perfusion imaging (LDPI). Mice were sacrificed at 28 days and tibialis anterior collected for histology.
Results:
LDPI of the ischemic hindlimb showed a return to baseline perfusion in WT mice (Figure). P2Y
2
KO mice recovered to <40% of baseline. Additionally, 4/8 P2Y
2
KO mice developed wounds of the ischemic limbs vs. 0/8 of WT mice. Histological analysis of tibialis anterior demonstrated regenerating myocytes in 3/4 P2Y
2
KO vs. 1/4 WT animals. By immunohistochemistry, angiogenesis was

significantly increased in P2Y
2
KO with a CD31+cell/myocyte ratio of 1.974 vs. 1.314 of WT (p=0.05). There was also greater inflammation in P2Y
2
KO with CD45+ cell infiltration over 3-fold that of WT (p<0.001).
Conclusion:
The P2Y
2
nucleotide receptor is important for blood flow recovery in a murine model of FAL. P2Y
2
KO mice also demonstrated delayed muscle regeneration with greater angiogenesis and inflammation in ischemic limbs. Further molecular analyses are needed to elucidate the mechanisms involved.
Abstract 281: Resveratrol Enhances Cholesterol Efflux Through PPAR-γ--Dependent Signaling Pathways in Human Macrophages
Authors: Hai Ofek Y , Voloshyna Iryna , Littlefield Michael J , Carsons Steven , Reiss Allison B
Published: July 2021

Introduction:
We previously reported that resveratrol, a plant derived polyphenol with numerous cardioprotective properties, improves cholesterol homeostasis by upregulating the expression of reverse cholesterol transport (RCT) proteins critical for preventing lipid overload and macrophage foam cell formation. However, the mechanism(s) by which resveratrol exerts this effect are unclear. The present study explores a possible mechanism for the atheroprotective actions of resveratrol on cholesterol metabolism.

Hypothesis:
We hypothesize that the anti-atherogenic effects of resveratrol are mediated through PPAR-γ dependent signaling pathways leading to upregulation of RCT proteins and enhanced cholesterol efflux.

Methods:
THP-1 macrophages, a pertinent model for atherosclerosis, were incubated for 20h with resveratrol (10μM) with/without the specific PPAR-γ antagonist GW6992 (1μM). Alternatively, cells were transfected with 100 nM of human PPAR-γ small interfering RNA (siRNA) for gene silencing prior to resveratrol treatment. Total RNA was isolated, converted to cDNA, and evaluated by QRT-PCR. Each reaction was done in triplicate. Western blot analysis was performed to confirm results of gene expression.

Results:
Resveratrol significantly increased expression of PPAR-γ, ABCA1, and 27-hydroxylase mRNA (mean±SEM, 136.2±8.5%, 168.3±9.2%, and 171.7±11.0% of control respectively, P<0.001). PPAR-γ gene silencing and PPAR-γ antagonist GW6992 effectively reduced PPAR-γ message by 90.4% and 54.7%, respectively (P<0.001). Both pharmacological blockade and gene knockdown of PPAR-γ nullified resveratrol effects on cholesterol efflux proteins. In cells treated with GW6992, mRNA levels of ABCA1 and 27-hydroxylase were decreased to 66.1±3.3% and 55.0±2.9% of control, respectively (P<0.001). Similarly, PPAR-γ silencing resulted in downregulation of ABCA1 and 27-hydroxylase expression to the level of control, which is significantly lower than in resveratrol treated cells (P<0.001). Data from gene expression studies were subsequently confirmed by Western blot analysis.

Conclusions:
We propose here a mechanistic model for the atheroprotective effects of resveratrol. Our data strongly suggests that resveratrol regulates cholesterol efflux and intracellular cholesterol processing via the PPAR-γ/LXR-α pathway.
Abstract 286: Phytochemical Treatment of Insulin Resistance in Metabolic Syndrome Patients
Authors: Siegel Günter , Knes Otto , Just Sören , Winkler Karl , Ermilov Eugeny
Published: July 2021

Background:
Since HOMA-IR predicts coronary artery calcification incidence and progression, but not independently of metabolic syndrome (MS), we sought to determine whether insulin resistance (IR) could be lowered by a phytochemical approach. In a 2-month clinical pilot study with Ginkgo biloba (EGb 761, 2 × 120 mg/d) on 11 MS patients, a novel biomarker spectrum embracing parameters of plaque formation, stability and progression, oxidative stress, inflammation, lipid composition and second messengers, made point-of-care theranostics feasible.

Methods:
Laser ellipsometry, photometric methods, ELISAs and EIAs were applied.

Results:
Nanoplaque formation was reduced by 14.3% (p < 0.01), nanoplaque size by 23.4% (p < 0.0004), oxLDL/LDL 21.0% (p < 0.002), 8-
iso
-PGF
2α
39.8% (p < 0.003), MPO 29.6% (p < 0.01), IL-6 12.9% (p < 0.04), hs-CRP 39.3% (p < 0.005), Lp(a) 26.3% (p < 0.001), MMP-9 32.9% (p < 0.04), whereas SOD was augmented 17.7% (p < 0.01), GPx 11.6% (p < 0.001), cAMP 43.5% (p < 0.001), and cGMP 32.9% (p < 0.001). Since none of the patients had a diabetes, we evaluated IR. Fasting morning glucose was reduced by 4.0% from 94.4 to 90.2 mg/dL (p < 0.03), insulin 8.6% from 12.7 to 11.5 mU/L (p < 0.04), and HOMA-IR 12.3% from 3.07 to 2.64 mU/L×mg/dL (p < 0.02). Because ginkgo is not an antidiabeticum, we looked out for a mechanistic explanation. Insulin (1), glucose (2) and HOMA-IR (3) were correlated to IL-6 (4), hs-CRP (5), TNFα (6) and TGFβ
1
(7), which changes in cytokine pattern could unravel lowering of the former quantities: 1 vs 4 (r = 0.73, p < 0.06); 2 vs 4 (r = 0.80, p < 0.03); 3 vs 4 (r = 0.71, p < 0.07); 1 vs 5 (r = 0.64, p < 0.03); 2 vs 5 (r = 0.75, p < 0.02); 3 vs 5 (r = 0.68, p < 0.14); 1 vs 6 (r = -0.68, p < 0.05); 2 vs 6 (r = -0.73, p < 0.04); 3 vs 6 (r = -0.67, p < 0.05); 1 vs 7 (r = -0.75, p < 0.09); 2 vs 7 (r = -0.70, p < 0.02) and 3 vs 7 (r = -0.89, p < 0.01).

Conclusion:
Ginkgo had beneficial effects on a multitude of arteriosclerotic and diabetic biomarkers. IR was diminished by 12.3% to an HOMA-IR markedly below 3 on average, a clinically tolerable level. Further, this decrease can explain the reduction in Ca
2+
-dependent nanoplaque formation and size. Thus, ginkgo may be used as complementary drug in the treatment of MS, arteriosclerotic and diabetic patients.
Abstract 293: Id3 Regulates High-Fat-Diet--Induced IgG2c Production
Authors: Harmon Daniel B , Oldham Stephanie N , Erickson Loren D , McNamara Coleen A
Published: July 2021

Background:
High-fat diet (HFD)-induced adipose tissue inflammation leads to insulin resistance and glucose intolerance, increasing the risk of cardiovascular disease. Recently, B cells and IgG antibodies have been shown to promote the metabolic consequences of obesity. HFD induces IgG2c production - an IgG isotype that promotes inflammatory disorders. Mice null for Id3, a helix-loop-helix protein known to mediate IgG2c responses
in vivo,
develop significantly attenuated HFD-induced obesity; raising the interesting hypothesis that Id3 promotes HFD-induced obesity by mediating HFD-induced IgG2c production.

Methods and results:
Western Blot analysis revealed that primary splenic B cells stimulated with IFN + CD40L (an IgG2c-inducing cocktail) expressed significantly greater amounts of Id3 protein compared to B cells treated with CD40L alone ((1.26+/-0.05 vs 0.88+/-0.03; n=3; p=0.003). ELISA determination of immunoglobulin levels from whole adipose tissue of fed C57Bl/6J (WT) mice fed a HFD (60% kcal fat) revealed a 4 fold increase in IgG2c (3810+/-789 ng vs 993+/-422 ng; n=5; p=0.01) - but not IgM, IgG1, or IgG2b - compared to chow-fed controls. No differences in serum IgG2c were observed. In contrast, HFD-induced adipose tissue IgG2c levels were significantly attenuated in Id3
-/-
mice (1643+/-493 vs 889+/-272 ng; n=6-7; p=0.19), despite the fact that Id3
-/-
mice had significantly more adipose tissue B cells per (g) fat compared to WT (1.6E5+/-0.2E5 vs 0.5E5+/-0.09E5; n=5; p=0.001).

Conclusions:
Id3 promotes HFD-induced local adipose tissue IgG2c production; an effect not dependent on B cell number. Studies in animals with B cell-specific loss of Id3 are ongoing to identify the mechanisms whereby Id3 mediates HFD-induced IgG2c production and downstream metabolic effects.
Abstract 319: Dalcetrapib-Mediated Modulation of Cholesteryl Ester Transfer Protein Activity Increases HDL-Cholesterol, Apolipoprotein A-I Levels and Cholesterol Efflux Capacity in Chow-Fed Rabbits
Authors: Brodeur Mathieu R , Rhainds David , Charpentier Daniel , Mihalache-Avram Téodora , Maugeais Cyrille , Rhéaume éric , Niesor Eric J , Tardif Jean-Claude
Published: July 2021

Introduction:
A potential approach to reduce CV risk is to increase HDL-C levels. This could be achieved by reducing cholesteryl ester transfer protein (CETP) activity. Dalcetrapib, which modulates CETP activity by changing its conformation and raises HDL-C without inhibiting CETP-induced pre-β-HDL formation in humans, was shown to decrease progression of atherosclerosis in rabbits.

Hypothesis:
Investigate the modifications of HDL particle size distribution and cholesterol efflux capacity of serum produced by dalcetrapib in normocholesterolemic rabbits.

Methods:
New Zealand white rabbits were treated with dalcetrapib (300 mg/kg as food admix) or placebo for 14 days. We evaluated CETP conformation and mass by ELISAs (including antibodies sensitive to conformational change), CETP activity by fluorescent lipid transfer, lipid profile and apoA-I distribution in HDL subclasses by 2D-non denaturing gradient gels (2D-NDGGE). Cholesterol efflux capacity of rabbit sera was determined after loading cells with
3
H-free cholesterol, using HepG2 hepatocytes to measure SR-BI-dependent efflux and by inducing ABCA1 or ABCG1 expression in BHK cells.

Results:
Dalcetrapib modified the conformation of rabbit CETP
in vitro
and
in vivo
and, after 14 days, this was associated with increased CETP mass (+50%, p<0.001) and reduced CETP activity (-86%, p<0.001). Total cholesterol was increased with dalcetrapib (+178%, p<0.001), due to a higher HDL-C level. In contrast, dalcetrapib reduced LDL-C and triglycerides by 41% (p<0.01) and 48% (p<0.001). Serum analysis by 2D-NDGGE showed that total rabbit apoA-I was increased 1.7- fold in animals treated with dalcetrapib. This was associated with an increase in large HDL but also in small α-migrating HDL with pre-β-HDL size. Cholesterol efflux assays showed that ABCA1-, ABCG1- and SR-BI-dependent efflux were all increased in dalcetrapib-treated rabbits (+24%, p=0.038; +21%, p=0.021; +44%, p<0.001).

Conclusion:
Modulation of CETP activity and conformation by dalcetrapib increases HDL-C and apoA-I levels and affects apoA-I distribution in HDL subclasses. These changes are associated with increased cholesterol efflux capacity, suggesting that HDL functionality is preserved in dalcetrapib-treated chow-fed rabbits.
Abstract 327: SR-BI Deficiency and Hypercholesterolemia Synergistically Lead to Impaired Erythropoiesis
Authors: Song Zhiqing , Guo Ling , Li Xiang-An
Published: July 2021

Recent studies showed that mice deficient in scavenger receptor BI (SR-BI) exhibit impaired erythropoiesis and it has been proposed that the high cholesterol environment plays a major role in the abnormal erythropoiesis. In this study, we utilized three animal models to assess the role of SR-BI and hypercholesterolemia in erythropoiesis. First, we used a high fat diet-induced hypercholesterolemia model. High fat diet caused a 2-fold increase in plasma cholesterol levels in SR-BI
+/+
(172 mg/dl to 350 mg/dl) and SR-BI
-/-
mice (313 mg/dl to 716 mg/dl). The high fat diet treatment markedly exacerbated the impaired erythropoiesis in SR-BI
-/-
mice as shown by 4-fold increase in reticulocyte percentage and 2.5-fold increase in early-to-late erythroblast ratio. Unexpectedly, high fat feeding did not induce abnormal erythropoiesis in SR-BI
+/+
mice despite of hypercholesterolemia in these mice. We then used SR-BI/LDLR double knockout mice to further elucidate the contribution of hypercholesterolemia to erythropoiesis. SR-BI/LDLR double knockout mice had 2-fold increase in plasma cholesterol levels and exhibited severer impaired erythropoiesis, similar to high fat-fed SR-BI
-/-
mice. Interestingly, despite of hyperlipidemia, LDLR single knockout mice did not display impaired erythropoiesis. Finally, we investigated the contribution of hepatic SR-BI using ScarbI
I179N
mutant mice, whose hepatic SR-BI expression has been knocked down by 90% and therefore has a 1.7-fold increase in plasma cholesterol levels compared to wild type controls. ScarbI
I179N
mice displayed normal erythropoiesis, similar to wild type controls. These findings indicate that hypercholesterolemia does not cause abnormal erythropoiesis in the presence of SR-BI, but markedly impairs erythropoiesis in the absence of SR-BI. We conclude that SR-BI is essential for normal erythropoiesis, and that hypercholesterolemia and SR-BI deficiency synergistically exacerbated impaired erythropoiesis.