bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 16: BcrKinase is a Novel Akt Kinase That Modulates Scavenger Receptor BI- and PDZK1-dependent Actions of HDL in Endothelium
Authors: Sacharidou Anastasia , Lee Wan-Ru , Shaul Philip E , Mineo Chieko
Published: June 2021

High density lipoprotein cholesterol (HDL) has direct atheroprotective actions on endothelium. These are mediated by scavenger receptor class B, type I (SR-BI) and its adaptor protein PDZK1, and they entail the activation of Akt kinase, which phosphorylates and thereby stimulates endothelial nitric oxide synthase (eNOS). In the present work we sought to determine how PDZK1 couples HDL/SR-BI to Akt and eNOS to modulate endothelial function. Using tandem affinity purification (TAP) following the infection of the human endothelial cell line EAhy926 with adenovirus expressing TAP-tagged PDZK1, we identified Breakpoint Cluster Region (Bcr) kinase as a PDZK1 interacting protein in endothelium. Whereas Bcr is well-known as a component of the Bcr-Abl fusion protein that results from translocation of the Philadelphia chromosome in chronic myelogenous leukemia, little is known of its function in endothelial cells. Bcr contains several distinctive domains including a C-terminal PDZ binding motif and a serine/threonine protein kinase domain. In primary human aortic endothelial cells (HAEC), we determined that endogenous Bcr interacts with PDZK1 in an HDL-dependent manner, and that Bcr is required for HDL-induced activation of eNOS and HDL stimulation of endothelial cell migration, which underlies the ability of the lipoprotein to promote endothelial monolayer integrity. Studies of mutant forms of Bcr with disruption of PDZK1 binding or kinase activity introduced into endothelial cells further revealed that Bcr-PDZK1 interaction and its kinase function are required for HDL activation of Akt and eNOS. Using a novel kinase assay that we recently developed that employs time-resolved Forster resonance energy transfer, we found that via SR-BI and PDZK1, HDL stimulates Bcr kinase activity in endothelial cells more than 20-fold. In addition, using Akt-based peptides in studies of the two known kinases for Akt, mTOR and PDK1, we determined that HDL activates Bcr kinase to directly phosphorylate Akt-Ser473 in an mTOR independent manner, and that Akt-Thr308 is a direct substrate of PDK1. These collective findings have identified Bcr to be a novel kinase for Akt, and they have revealed that Bcr is critically involved in HDL modulation of endothelial cell phenotype.
Abstract 182: Inhibitor of differentiation (Id)3 Regulation of the Vasculature in Adipose Tissue
Authors: Osinski Victoria , Kirby Jennifer , Sonkusare Swapnil , Civelek Mete , McNamara Coleen
Published: June 2021

Rationale:
Over half of the U.S. is overweight or obese and excessive weight gain greatly increases one’s risk of cardiovascular disease. Adipose tissue requires new blood vessel formation to support transport of nutrients during expansion. Our lab has previously published that global loss of the helix-loop-helix transcription factor Inhibitor of differentiation 3 (Id3) results in attenuated increases in adipose depot size and microvascular blood volume in mice fed a high fat diet (HFD). Id3 has also been implicated in regulating tumor angiogenesis. Together, our results suggest that Id3 is regulating HFD-induced angiogenesis in adipose tissue.

Methods and Results:
To determine if Id3 is regulating angiogenesis, we performed genetic, cellular, and arterial experiments and analyses. Using bromodeoxyuridine (BrdU) treatments and flow cytometry, we found that endothelial cells within adipose tissue of mice with a global genetic knockout of Id3 (Id3 KO) had fewer proliferating (BrdU-positive) endothelial cells (0.0679% BrdU+CD31+ cells) than wild type (WT) mice (0.754% BrdU+CD31+). Using mouse adipose tissue gene expression data from the Gene Ontology Consortium database, we found that genes correlating most significantly with Id3 were functionally classified under categories of vascular development (p = 4.45E-8, correlation significance) and angiogenesis (p = 1.66E-6). Sox18, a known transcriptional regulator of blood vessel formation, shows higher relative gene expression in Id3 WT (1.42 ± 0.417) than Id3 KO (0.332 ± 0.126) adipose tissue. Finally, preliminary findings using pressure myography demonstrate that Id3 KO mice have a higher baseline arterial diameter than Id3 WT.

Conclusion:
Dysregulation of vascular development during adipose tissue expansion is altering arterial function in an Id3-dependent manner.
Abstract 186: Hdac7-derived 7aa Peptide May Function as a Phosphorylation Carrier
Authors: Yang Junyao , Wang Wen , Wang Qian , Zeng Lingfang
Published: June 2021

Background:
Histone deacetylase 7 (HDAC7) belongs to class II HDAC family, playing a pivotal role in the maintenance of endothelium integrity. There are 8 splicing variants in mouse HDAC7 mRNAs. Within the 5’ terminal non-coding area of some variants, there exist some short open reading frames (sORFs). Whether these sORFs can be translated and their potential roles in cellular physiology remain unclear.

Method and results:
Our previous studies suggested that one mouse HDAC7 produced a 7aa peptide from the non-coding area. In this study, we demonstrated that one sORF encoding a 7 amino acids (aa)-peptide could be translated in response to vascular endothelial cell growth factor (VEGF) in vascular progenitor cells (VPCs). The 7aa-peptide (7A) could be phosphorylated at serine residue via MEKK1. Importantly, the phosphorylated 7aa-peptide (7Ap) could transfer the phosphorylation group to the Thr residue of the 14-3-3γ protein in a cell free in-gel buffer system. The in vitro functional analyses revealed that 7A enhanced VEGF-induced VPC migration and differentiation toward endothelial cell (EC) lineage, in which MEKK1 and 14-3-3γ served as upstream kinase and downstream effector respectively. Knockdown of either MEKK1 or 14-3-3γ attenuated VEGF-induced VPC migration and differentiation. Exogenous 7Ap could rescue VEGF effect in MEKK1 but not in 14-3-3γ knockdown cells. The in vivo studies showed that 7A especially 7Ap induced capillary vessel formation within matrigel plug assays, increased re-endothelialization and suppressed neointima formation in the femoral artery injury model, and promoted the foot blood perfusion recovery in the hindlimb ischemia model.

Conclusion:
These results indicate that the sORFs within the non-coding area can be translated under some circumstances and that the 7aa-peptide may play an important role in cellular processes like migration and differentiation via acting as a phosphorylation carrier.

Significance:
As a phosphorylation carrier, 7aa possesses therapeutic potentials in tackling angiogenesis related diseases.
Abstract 197: A Novel Association Between High Density Lipoprotein Levels and the Risk of Postoperative Acute Kidney Injury
Authors: Smith Loren E , Smith Derek K , Linton MacRae F , Billings Frederic T
Published: June 2021

Acute kidney injury (AKI) after cardiac surgery occurs in up to 30% of patients and is an independent predictor of death. HDL may attenuate mechanisms of AKI. We hypothesized that a high preoperative HDL cholesterol concentration is protective against postoperative AKI.

We analyzed data from a prospective, 393-subject trial of perioperative atorvastatin to prevent post-cardiac surgery AKI. Statin-using patients were randomized to placebo or 80mg atorvastatin the morning of surgery and 40mg on postoperative day 1. Stain-naïve patients were randomized to placebo or 80mg the day prior to surgery and 40mg daily thereafter during hospitalization. The association between HDL level and maximum serum creatinine change from baseline in the first 48 postoperative hours was assessed using a two-component latent variable mixture model and AKI risk factors. Regression analyses assessed interactions of chronic statin use, perioperative atorvastatin treatment, and HDL level on AKI risk.

Postoperative AKI occurred in 99 patients (25.2%). Median (10
th
, 90
th
percentile) preoperative HDL was 37.6 (25.0, 54.0) mg/dL and postoperative creatinine change 0.09 (-0.11, 0.59) mg/dL. Lower HDL levels were independently associated with increased creatinine rise (p=0.02) (Figure 1A). Regression analysis showed this association was present in statin-using but not statin-naïve patients (p=0.008) (1B). The protective effect of high HDL in chronic statin users was enhanced with perioperative atorvastatin treatment (p=0.004) (1C) and with increasing chronic statin dose (p=0.003) (1D). Similar analyses using LDL found no association with AKI risk (p=0.51).

Conclusions:
Higher preoperative HDL was associated with less risk of AKI. Statin exposure modified this association. Specifically, subjects with higher HDL levels on chronic statin therapy had less creatinine rise and appeared to further benefit from higher chronic statin dose and perioperative atorvastatin therapy.
Abstract 201: Maternal HDL-Cholesterol Levels in Women From the Gambia are Directly Related to Infant Birthweight
Authors: Rebholz Sandra L , Melchior John T , Welge Jeff , Prentice Andrew M , Moore Sophie E , Davidson Sean , Woollett Laura A
Published: June 2021

Babies born with low birthweight are often at a health disadvantage. Previous studies have shown direct relationships between maternal plasma cholesterol and infant birthweight in resource-rich countries. As plasma cholesterol levels are often decreased in resource-poor countries, the purpose of these studies was to evaluate the relationship between plasma cholesterol and birthweight in women enrolled in the ENID trial (ISRCTN49285450) in rural Gambia, West Africa. Plasma was obtained at enrolment (13.6±3.3 wk) and at 20 and 30 weeks of gestation; samples were obtained from women with term infants that weighed <2.75 kg or >3.25 kg at birth. Women with lower HDL-cholesterol (HDL-C) concentrations in mid-pregnancy had lower birthweight infants compared to women with higher HDL-C concentrations. There was no significant association between LDL-C or total cholesterol concentrations and birthweight. The relationship with HDL-C and birthweight was maintained when maternal BMI was included in the model. To begin to elucidate the processes involved in the regulation of fetal growth, placental function was examined in mice with increasing maternal HDL-C concentrations based on apoA-I levels; mice were lacking apoA-I (apoA-I
-/-
), were wildtype (apoA-I
+/+
), or had excess apoA-I (apoA-I
tg/tg
). HDL decreased in size as plasma apoA-I levels increased, and there were no statistical differences in the proteins carried by HDL, except for apoA-I, in pregnant mice of different genotypes. However, pregnancy alone led to changes in the HDL proteome. Importantly, fetuses of mice with lower concentrations of maternal HDL-C had reduced growth rates, not due to a lack of fetal apoA-I. The murine fetal growth rates were directly related to nutrient uptake by and transport across the placenta. This work suggests that maternal HDL affects placental function leading to enhanced nutrient supply and improved growth in utero, making HDL a potential biomarker for fetal growth and putative target for intervention.
Abstract 207: Wound Application of Nitrite Enhances Healing in Diabetic Mice
Authors: Aggarwal Ankur , Madigan Michael C , Cody Ellen , Tzeng Edith
Published: June 2021

Background:
Impaired diabetic wound healing, a source of significant disability, has been attributed to the reduced nitric oxide (NO) bioavailability and increased oxidative stress. We have previously reported that xanthine oxidoreductase (XOR), important in purine metabolism and reactive oxygen species synthesis, is highly expressed in regenerating skin and is required for normal healing. XOR is also expressed in diabetic wounds at similar or greater levels. Recently, XOR has been identified as a strong nitrite reductase and can generate NO from nitrite. Based on these findings, we hypothesize that topical nitrite applied to diabetic wounds may improve healing.

Methods:
Anesthetized diabetic db/db mice underwent excisional wounding. The wounds were treated with either Aquaphor with 2% sodium nitrite or Aquaphor alone and covered with a bioocclusive dressing. Wounds were photographed every other day for digital planimetry until day 28. Wounds were collected on day 28 and fixed, sectioned, and examined for proliferation (Ki67) and angiogenesis (CD31) by immunostaining.

Results:
Wounds in db/db mice treated with topical Aquaphor + nitrite displayed significantly accelerated healing compared to those treated with Aquaphor alone (Table). Aquaphor + nitrite also increased wound angiogenesis (29.3 ± 4.7 vs 13.2 ± 4.8, N=6-8/group; p<0.038) and keratinocyte proliferation (20.4 ± 5.1 vs 3.9 ± 2.4; p<0.034) compared with those treated with Aquaphor alone.

Conclusion:
Topical nitrite significantly accelerated wound healing in db/db mice. Nitrite treatment enhanced angiogenesis and keratinocyte proliferation in the wounds. These prohealing effects are likely mediated by XOR mediated conversion of nitrite to NO which can potentially divert XOR away from ROS production. Future studies will define the ability of nitrite to alter wound oxidative stress as well as the translation of this simple and safe therapy for human application.
Abstract 221: Anti-angiogenic Therapy Rescues the Defect in Ischemia-Induced Neovasculature in Type II Diabetic Mice
Authors: Wu Jianbo , Xiao Lamei , Yang Yan , Deng Xin , Pang Ningbo
Published: June 2021

Objective:
Type II diabetes mellitus (DM) leads to the development of microvascular diseases and is associated with impaired angiogenesis. The presence of VEGF can block PDGF-dependent regulation of neovascularization and vessel normalization. We tested the hypothesis that the inhibition of VEGF rescues the defect in Ischemia-Induced neovasculature by inducing PDGF-BB activation.

Methods and Results:
Hindlimb ischemia was generated by femoral artery occlusion in diet-induced diabetic mice and age-matched normal chow (NC)-fed mice. Bevacizumab, a humanized monoclonal antibody against VEGF-A, was intraperitoneally administered to mice (200 μg/mouse; once weekly for 2 weeks) to determine if bevacizumab restores neovasculature formation in DM mice. Color Laser Doppler showed that the blood flow perfusion in ischemic hindlimb was significantly increased in the DM treated by bevacizumab compared to vehicles at 7, 12, and 14 days (n=6/group, p<0.05). Real-time PCR revealed that the expression of PDGF-BB was significantly enhanced in the DM treated by bevacizumab compared to vehicles. Furthermore, anti-PDGF drug significantly inhibit the blood flow perfusion in the DM mice treated by bevacizumab.

Conclusions:
These results indicate that inhibition of VEGF could restore PDGF-BB expression, and possibly imply the molecular target of diabetic foot.
Abstract 224: Biological Clock Strongly Regulates Brown Adipose Tissue Activity: Implications for Postprandial Triglyceride Metabolism
Authors: Berbée Jimmy , van den Berg Rosa , Kooijman Sander , Ramkisoensing Ashna , Coomans Claudia P , Meijer Johanna H , Biermasz Nienke R , Rensen Patrick C
Published: June 2021

Background:
Brown adipose tissue (BAT) is an attractive anti-dyslipidemia target as it burns high amounts of triglycerides (TG) into heat. In humans, plasma TG levels display 24 h variation independent of food intake, indicating a strong regulation by the biological clock. We recently showed that prolonged day length decreases the uptake of TG by BAT. We now aimed to assess the 24 h rhythm of BAT activity, the effect of day length thereon, and the consequences for postprandial TG metabolism.

Methods and Results:
Male C57Bl/6J mice were exposed to short (8 h), regular (12 h), or long (16 h) days during 5 weeks. The rhythm of BAT was determined by its capacity to take up plasma TG-derived fatty acids (FA) at 6 time points during a 24 h period. Mice exposed to regular days displayed a remarkable diurnal pattern of TG-derived FA uptake from glycerol tri[
3
H]oleate-labeled VLDL-like particles by BAT, but not by other tissues, reaching a peak [
3
H]FA uptake at onset of dark. Short day length increased the amplitude of [
3
H]FA uptake by 3-fold and advanced the peak of [
3
H]FA uptake, again to onset of dark. Conversely, long day length delayed the peak to onset of dark. We next assessed whether BAT rhythmicity affects postprandial TG clearance in dyslipidemic APOE*3-Leiden.CETP mice. Mice were fed a western-type diet that were exposed to either a short (8 h) or long 16 h (long) days and gavaged with olive oil at several time points during the day. Irrespective of day length, postprandial TG excursion was virtually absent before the onset of dark (highest BAT activity) and high before onset of light (lowest BAT activity).

Conclusion:
Day length dictates a diurnal rhythm in TG-derived FA uptake capacity of BAT, thereby strongly determining postprandial TG metabolism. We propose that the diurnal variations in TG levels observed in humans may be explained by a diurnal rhythm in BAT activity.
Abstract 234: Reduction of LpX and Improvement in Kidney Function in LCAT-KO Mice by Enzyme Replacement Therapy
Authors: Vaisman Boris L , Neufeld Edward B , Freeman Lita A , Sampson Maureen L , Pryor Milton , Axley Milton J , Karathanasis Sotirios K , Remaley Alan T
Published: June 2021

Familial LCAT Deficiency (FLD) is associated with the gradual development of severe kidney dysfunction from the renal deposition of LpX, an abnormal lipoprotein that accumulates in this disorder. The aim of our study was to develop an efficient mouse model of FLD in which elevated plasma LpX and kidney dysfunction can be rapidly induced in order to test whether recombinant human LCAT (rhLCAT) injections can prevent renal disease. We used the previously described LCAT-Ko x SREBP1a transgenic mouse model (Zhu et al., 2004) with the transgene placed under control of PEPCK promoter, which can be induced by high protein diets. We demonstrate that high levels of LpX plasma particles appeared within 5-7 days after 2-3 month old mice were switched to a protein rich carbohydrate low diet (PRCL). Kidney dysfunction measured by albumin/creatinine ratio after 9 days on PRCL diet increased by 32±12 fold. On the PRCL diet, plasma VLDL-C and LDL-C fractions of LCAT-Ko x SREBP1a mice increased 2-4 fold compared to mice on a normal chow diet. Transmission electron microscopy clearly demonstrated the presence of multilamellar LpX particles in plasma and renal glomeruli, as well as robust accumulation lipid droplets in hepatocytes of mice kept on the PRCL diet for 7 days. All these changes were reversible; when mice were returned to the normal chow diet their plasma lipid characteristics and kidney function quickly returned within days to the level observed before the diet was initiated. When LCAT-Ko x SREBP1a mice were placed on PRCL diet and simultaneously treated with rhLCAT (x 3 per week IV injection 30 mg/kg for 2 weeks) plasma LpX was eliminated and a normal mouse lipoprotein profile was observed and most notably HDL-C increased from 5.6 g/dL to 54.1 mg/dL. Treatment with rhLCAT decreased the albumin/creatinine ratio 5±1 fold.

Conclusions:
LCAT-Ko and SREBP1a mice on a PRCL diet can be used as efficient model for investigating potential therapies for LCAT deficiency. Results show the feasibility of rhLCAT treatment for preventing renal disease in patients with FLD.
Abstract 249: Increased Vascular CCRL2 Expression by Disturbed Blood Flow Promotes Atherosclerotic Plaque Formation in ApoE Deficient Mice
Authors: Hu Shuhong , Zhu Li
Published: June 2021

Atherosclerosis is a chronic inflammatory disease of the arterial wall elicited by accumulation of LDL and leucocytes in the subendothelium at predilection sites with disturbed laminar flow. Chemokines and their receptors appear to act as critical players in atherosclerosis as they not only direct atherogenic recruitment of leucocytes but also exert cell hemostatic functions by chemokine ligand-receptor axes and their specific or combined contributions. Atypical chemokine (C-C motif) receptor-like 2 (CCRL2) cooperates with its ligand chemerin and leukocyte-expressed chemerin receptor chemokine-like receptor 1 (CMKLR1) to regulate cell trafficking and inflammatory responses,but its role in atherosclerosis is not clear. To investigate whether CCRL2 contributes to the pathomechanism of atherogenesis, we generated CCRL2
-/-
mice in hyperlipidemic atherosclerosis-prone ApoE
-/-
background and found that the atherosclerotic plaque area of the total aorta was significantly reduced compared with CCRL2
+/+
ApoE
-/-
mice on a high fat diet. The protective effect of CCRL2 deficiency was anatomically isolated primarily to the site of disturbed blood flow (D-flow) in the aortic arch but not in the descending aorta. Endothelial CCRL2 was upregulated in response to D-flow and either CCRL2 or CMKLR1 deletion reduced plaque formation. Further studies showed that CCRL2 co-localized with CMKLR1 and chemerin within the atherosclerotic aorta root. CCRL2 deficiency led to significantly less lipid deposition in aortic root, reduced CMKLR1
+
leukocyte rolling on lesional vascular endothelium, diminished macrophage accumulation and foam cell formation, and polarized macrophage to an M2-like phenotype. These results demonstrate that D-flow induction of vascular CCRL2 is required for optimal formation of atherosclerotic plaques via coordinating the accumulation of CMKLR1
+
monocytes/macrophages within the vascular wall, and thus identifies CCRL2 as a novel drug target to prevent or treat atherosclerosis.

This work was supported by Natural Science Foundation of China (grant 81370373 to L.Z. and 31300781 to C.T.)

Key Words:
atherosclerosis, CCRL2, chemerin, macrophage
Abstract 25: Inducible Depletion of Calpain-2 Attenuates Angiotensin II-induced Cytoskeletal Structural Protein Destruction During Abdominal Aortic Aneurysm Development in Mice
Authors: Muniappan Latha , Javidan Aida , Jiang Weihua , Moorleghen Jessica J , Balakrishnan Anju , Howatt Deborah A , Subramanian Venkateswaran
Published: June 2021

Background and Objective:
In abdominal aortic aneurysm (AAA) patients, structural integrity of the aortic wall is disrupted due to the dissociation of cytoskeletal structural proteins that bridge SMC contractile filaments with extracellular matrix by proteases. However, identity of functional proteases which target cytoskeletal structural linker proteins remain unknown. Recently, using the angiotensin II (AngII)-induced AAA model, we demonstrated that AngII significantly increased fragmentation of cytoskeletal structural protein, Filamin A, in AAAs. Further, using a pharmacological inhibitor and genetic deficient mice, we identified that calpain-2 (a class of calcium-activated, intracellular cysteine proteases) plays a critical role in AngII-induced AAA formation in mice. The purpose of this study was to determine the functional contribution of calpain-2 in AngII-induced cytoskeletal structural protein destruction during AAA development.

Methods and Results:
Calpain-2 floxed mice that were hemizygous for Cre-ERT2 in an LDLr -/- background were produced by breeding male Cre-ERT2 to female calpain-2 floxed mice. At 8 weeks of age, male Calp-2 x Cre-ERT2 (Cre+) and non-Cre littermates (Cre-) mice were injected with tamoxifen (25 mg/kg, i.p.) daily for 5 consecutive days. After 2 weeks, Western blot analyses showed a complete depletion of calpain-2 protein in the aorta from Cre+ mice compared to Cre- littermates. Male Cre+ and Cre- (N=12 each) mice were fed a Western diet and infused with saline or AngII (1,000 ng/kg/min) by osmotic minipumps for 2 weeks. AngII infusion significantly (P<0.05 vs saline) increased c-terminal fragmentation of cytoskeletal structural protein, talin and kinases such as integrin linked kinase-1 (ILK-1), and focal adhesion kinase in addition to filamin A. Inducible depletion of calpain-2 significantly (P<0.05; Cre+ vs Cre-) blunted AngII-induced fragmentation of talin, filamin A and ILK-1. Further siRNA mediated silencing of calpain-2 in aortic SMCs and fibroblasts significantly reduced AngII-induced fragmentation of filamin A and talin.

Conclusion:
These findings suggest that calpain-2 plays a critical role in AngII-induced cytoskeletal structural protein fragmentation during AAA development in mice.
Abstract 270: Plant Based Diet Improves Glucose Metabolism in 18 Days
Authors: Ramirez Francisco E , Siebold Jill , Ivy Linda , Antuna Kathelyn , Sanchez Albert , Nedley Neil
Published: June 2021

Background:
Impaired glucose metabolism affects atherosclerosis. Improvement in blood glucose could prevent many future complications.

Hypothesis:
The low caloric density of plant-based diets could decrease weight affecting fasting blood glucose (FBG) levels

Methods:
Data from 1827 patients that received residential treatments at a lifestyle center in Weimar, CA was used. A low fat whole plant based diet was prescribe together with intensive exercise, sleep hygiene, medical therapy, hydrotherapy, massage, psychological and spiritual therapies were employed. A physician monitored the patients during their 18 days of training and intervention. Some patients reduced their blood glucose medication during the 18 days. The morning FBG was measured at baseline and at the end of 18 days. Mg/dl scale was used.

Results:
From 1827 patients, 1193 of them were females. Mean age at baseline was 59.6 SD 15.5. At baseline fasting blood sugar was (average, median, mode, min, max) 108.2, 96, 90, 39, 437, 39.5 SD 39, SEM .9. At the end of the 18 days it was measured at (average, median, mode, min, max) 102.1, 93, 89, 51, 495, SD 32, SEM .7. Weight and BMI during this time also changed. The average weight at the beginning was 178 lbs SD 54.7, SEM 1.2 and at end it was 174 lbs SD 51, SEM 1.2. Average BMI at beginning was 28.6 SD 7.7, and at end was 27.9 SD 7.7.

Conclusion:
Plant based diet, as well as lifestyle changes can help decrease FBG levels. Some patients had an increase in FBS due to decrease of their medication.
Abstract 271: Animal Products Tend to Increase Hemoglobin A1c Among Depressed Individuals
Authors: Ramirez Francisco E , Siebold Jill , Nedley Neil , Antuna Kathelyn , Sanchez Albert , Grilo Paulo
Published: June 2021

Background:
Development of a diabetic state has been correlated with the use of energy-rich foods. Depressed individuals are prone to have erroneous diet habits.

Hypothesis:
Animal products are energy-rich products. Intake of these products could affect glucose levels in depressed individuals.

Methods:
The study analyzed participants in a 10-day in-patient residential ,depression recovery program. A total of 309 participants that completed the program where studied: 106 men and 203 women. On day one, all participants completed a depression and anxiety assessment test, a validated 75-item self-report screening tool that determines (1) depression based on DSM-5 (Diagnostic and Statistical Manual of Mental Disorders) criteria, (2) anxiety level, (3) mini EQ (Emotional Quotient), (4) demographic data, and (5) dietary intakes. The fact that meat, cheese and fish were part of each participant’s diet was determined dichotomously using their answers on the assessment test. A potential diabetic state was screened in each participant by analyzing hemoglobin A1c level in day one of the program. The participants’ dietary habits were correlated to the hemoglobin A1c level. SPSS software was used to process the data. An independent-samples t-Test was conducted to compare the intake of various energy rich foods with hemoglobin A1c level.

Results:
At baseline participants had a mean Beck Depression index II of 27.4 (SD = 10.1, min = 7, max = 53), corresponding with high moderate depression. Results exposed that consumption of meat at least twice a day (M=5.5, SD=.68, SEM=.4) compared to no meat intake (M=5.7, SD=1.15,SEM=.17), was related with higher levels of hemoglobin A1c, t(286)=-2, p = 0.041. This association was not found for cheese t(286)=0.45, p = 0.65 or fish t(286)=0.9, p = 0.368 intake.

Conclusion:
It seems meat intake, excluding fish or chicken, has a negative effect in the level of hemoglobin A1c in a clinically depressed population. To further confirm these results, prospective studies are required, as to determine its mechanisms.
Abstract 301: Aortic Segmental Diameters are Influenced by Both Common and Segment-specific Factors
Authors: Chen Xiaofeng , Yang Xinan , Jiang Jianjun , Xu Baohui , Tang Lijiang
Published: June 2021

Background and Objective:
Aortic diameter is a critical parameter for the diagnosis of aortic aneurysm. In non-aneurysm patients, a large infrarenal aortic diameter are also a risk for all-cause mortality. Additionally, aortic dimension is associated with several risk factors of coronary artery disease; the measure of subclinical disease; a predictor of incident congestive heart failure; stroke risk; and all-cause and cardiovascular disease mortality. The purpose of this study was to explore the risk factors and potential alternative pathogenic mechanisms of aortic dilatation.

Methods and Results:
Five hundred and twenty patients with clinically evident arterial disease or cardiovascular risk factors were prospectively recruited. Comprehensive transthoracic M-mode, 2-dimensional, and Doppler echocardiographic studies were performed using commercially and clinical diagnostic ultrasonography technique. The aortic dimensions were assessed at end-diastole at the different levels: (1) the annulus, (2) the mid-point of the sinuses of Valsalva, (3) the sinotubular junction, (4) the ascending aorta at the level of its largest diameter, (5) the transverse arch, (6) the descending aorta posterior to the left atrium, and (7) the abdominal aorta just distal to the origin of the renal arteries. The relationships of aortic dimensions with clinical characteristics were assessed by linear multiple regression analysis. Variables analyzed included common cardiovascular risk factors, co-morbidities, lipid profile, hematological parameters. Age and diastolic blood pressure were positively associated with the diameters of arch, descending, ascending and abdominal aorta. In contrast, female gender and hypertension were negatively associated with the diameters of arch, descending, ascending and abdominal aorta. The presence of coronary arterial disease were positively associated with arch diameter, whereas High-density lipoprotein cholesterol levels were positively associated with the diameters of ascending, descending and abdominal aorta. Effects from other factors varied among aortic segments.

Conclusion:
Aortic segmental diameters are influenced by both common and segment-specific factors.
Abstract 31: Resolvin D1 Attenuates PDGF-induced Vascular Smooth Muscle Cell Migration via the cAMP Pathway
Authors: Mottola Giorgio , Wu Bian , Chatterjee Anuran , Chen Mian , Conte Michael S
Published: June 2021

Introduction:
Resolvin D1 (RvD1), a specialized pro-resolving lipid mediator (SPM), attenuates migration in vascular smooth muscle cells (VSMC), which is critical to the development of neointimal hyperplasia. SPM are known to interact with G-protein coupled receptors (GPCR). We sought to investigate the pathways by which RvD1 influences VSMC migration.

Methods:
VSMC were harvested from human saphenous veins. cAMP levels were measured via ELISA in the absence or presence of RvD1 receptor (ALX, GPR32) blockers. NF449, a G
s
-protein inhibitor, was also used. PDGF-BB (10ng/ml) was used as an agonist in a VSMC scratch assay as well as the receptor blockers. PDGF-BB-induced cytoskeletal changes were measured as aspect ratio after actin staining, and a scratch assay was used to assess migration. PDGF-induced Rac1 activity was measured via ELISA; VASP phosphorylation was assessed by Western blot and Paxillin translocation by Immunofluorescence. PKA inhibitor Rp-8-Br-cAMP (10μM) was used in the phenotypic and downstream signaling studies.

Results:
RvD1 treatment (10nM) of VSMC induced a significant acute flux in cAMP levels at 5 minutes (Fig. 1A; n≥3); this increase was abolished by an ALX antagonist (WRW4), the anti-GPR32 blocking Ab, and NF449. RvD1 (10nM) attenuated PDGF-induced VSMC migration (Fig. 1B-C; n≥3), cytoskeletal rearrangements (n=4), Rac1 activation (n≥3) and increased phosphorylation of VASP (n=3). These effects were negated by the addition of Rp-8-Br-cAMP, suggesting cAMP involvement. RvD1’s anti-migratory effect was reversed by blocking ALX or GPR32 (Fig. 1C; n≥8).

Conclusion:
Our results suggest that RvD1 attenuates VSMC migration by increasing levels of cAMP through ALX and GPR32 via a G
s
-protein-mediated action. Interference with Rac1, VASP and Paxillin function appear to be important for the anti-migratory activity of RvD1.
Abstract 317: Paracrine Effects of Satellite Cells on Collateral Vessel Formation
Authors: Hansen Laura , Joseph Giji , Weiss Daiana , Taylor W. Robert
Published: June 2021

Peripheral artery disease is a major health problem in the United States that effects 8.5 million people and can lead to limb pain, decreased mobility, and in severe cases amputation. The ability to form a robust collateral network to restore blood flow and prevent ischemia leads to a better prognosis and restoration of function. The growth of collaterals is a complex process that involves recruitment of various cell types including smooth muscle cells, endothelial cells, and macrophages. Migration and proliferation of these cells are processes regulated by numerous cytokine and paracrine signals. We hypothesize that an important and novel source of these signals is satellite cells. Satellite cells are myogenic progenitor cells that lie below the basal lamina of muscle fibers. In healthy muscle, the cells are quiescent but in response to injury, such as ischemia, they become activated and proliferate. We hypothesized that activated satellite cells produce factors that will influence critical cells for vessel formation in addition to differentiating to repair muscle. To study the paracrine effects of satellite cells on vascular smooth muscle cells, we used a co-culture system with freshly isolated satellite cells from the ischemic leg as the stimulus. We found that satellite cells significantly increased smooth muscle migration 2.5 fold compared to media alone using a modified Boyden chamber assay. BrdU staining to assess proliferation showed modest increases in smooth muscle proliferation (1.3 fold change, p<0.01). Finally, to investigate these paracrine effects in vivo, we delivered alginate encapsulated satellite cells to mice following the hind limb ischemia procedure, which is a model of collateral growth. We found that mice that received the encapsulated satellite cells had significantly improved perfusion as measured by Laser Doppler imaging at day 14 post surgery when compared to empty capsules (perfusion ratio of 0.87 ± 0.04 (cells) vs 0.68 ± 0.07 (empty capsules), p<0.05). This result demonstrates that satellite cells can positively influence collateral growth in vivo. We believe that satellite cells play a critical role in collateral vessel formation and may potentially be a therapeutic strategy for the treatment of peripheral artery disease.
Abstract 326: Group IIA Secretory Phospholipase A
2
Predicts Graft Failure and Cardiovascular Mortality in Renal Transplant Recipients by Mediating Decreased Kidney Function
Authors: Annema Wijtske , de Boer Jan Freark , Dikkers Arne , Bakker Stephan J , Tietge Uwe J
Published: June 2021

The acute phase protein group IIA secretory phospholipase A2 (sPLA2-IIA) has proatherosclerotic properties. The present study prospectively investigated whether plasma sPLA2-IIA levels are associated with graft failure, cardiovascular and all-cause mortality in renal transplant recipients (RTRs), patients known to be susceptible to accelerated atherosclerosis, both in the graft and in the systemic vasculature. In 495 RTRs (median follow-up 7.0 years) sPLA2-IIA was determined at baseline and was significantly higher in RTRs than in healthy controls (median 384 ng/dL [range 86-6951] vs. 185 ng/dL [range 104-271], P<0.001), but lower than in end-stage renal disease patients (median 1053 ng/mL [range 458-2599], P<0.001). Kaplan-Meier analysis demonstrated an increased risk for graft failure (P=0.002), cardiovascular (P<0.001) and all-cause mortality (P<0.001) with increasing gender-stratified quartiles of sPLA2-IIA. Cox regression analyses showed a strong association of sPLA2-IIA with increased risks of graft failure (hazard ratio=1.42[1.11-1.83], P=0.006), cardiovascular (hazard ratio=1.48[1.18-1.85], P=0.001) and all-cause mortality (hazard ratio=1.39[1.17-1.64], P<0.001). However, this association was largely explained by parameters of kidney function. Further analyses in RTRs demonstrated that patients with higher baseline sPLA2-IIA levels showed faster decline in renal function during follow-up. In addition, kidney function in human sPLA2-IIA transgenic mice deteriorated more rapid over time as compared with wild-type controls (urinary albumin:creatinine ratio at 48 weeks of age 774±156 vs. 193±60, P<0.01). In summary, this prospective study demonstrates that sPLA2-IIA is a significant predictive biomarker for the occurrence of chronic graft failure, overall and CVD mortality in RTRs dependent on kidney function. This dependency is explained by sPLA2-IIA impacting negatively on kidney function over time in humans and transgenic mice.
Abstract 331: Restless Leg Syndrome and Its Association with Iliofemoral Venous Disease
Authors: Reddy Navya
Published: June 2021

Background:
Restless leg syndrome (RLS) is one of the most common sleep disorders, affecting between 5% and 10% of the U.S. population.1 However, it has an unclear etiology, and is often over-medicated without a resulting qualitative improvement in lifestyle. It has not been determined whether iliofemoral or iliocaval venous disease resulting from either thrombotic lesions or external compression, has any association with RLS.

Methods:
A total of 510 patients treated for iliofemoral venous compression syndrome (IVCS) with angioplasty and stenting were documented to have concomitant RLS per NIH screening criteria,2 prior to endovenous revascularization. 367 of the following patients were selected for retrospective research analysis based on pre-procedural selection criteria which included: IVCS with > 60% luminal stenosis, confirmed RLS > 1 year with or without current treatment, diminished quality of life, and clinical signs and symptoms of chronic venous disease. Patients with Diabetes, End stage renal disease, current pregnancy, and Rheumatic disease were excluded from this study. Patients with thrombotic lesions of the iliofemoral veins were also excluded. Following revascularization, patients were followed at 6 weeks, 3 months, and 6 months with doppler evaluation and asked to rate the severity of their RLS using a scale developed by the International Restless Leg Syndrome study group.3

Results:
Of the 367 patients studied, 52% of patients had improvement of their RLS symptoms at 3 months, 90% reported significant relief with cessation of medication(s) within 6 months, and 10% did not have any statistically significant improvement by 6 months (p < 0.001).

Conclusion:
This study demonstrates a strong relationship between deep vein outflow obstruction, with either iliocaval or iliofemoral stenosis, and RLS. It is suspected that a large population suffers from iliofemoral venous disease, and including this subset of patients in the differential diagnosis for RLS can allow for earlier treatment of IVCS and prevention of RLS.
Abstract 335: Mechanical Valve Thrombosis and Therapy
Authors: Osoro Moses
Published: June 2021

Background:
Obstruction of prosthetic heart valves may be caused by thrombus formation and/or pannus in-growth. Thrombosis is more likely with inadequate anticoagulation and associated with more acute onset of symptoms.

Case:
A 36 year old female with a history of bicuspid aortic valve, now with a mechanical aortic valve (21mm Carbomedics Top-Hat) presented with sudden onset of exertional dyspnea and orthopnea (NYHA Class III). She had been compliant with anticoagulation, with bi-monthly INRs ≥2.5. On admission, her INR was 3.14. Physical exam revealed pedal edema and 3/6 crescendo-decrescendo murmur radiating to the carotids.

Transthoracic echocardiogram (TTE) showed a left ventricular ejection fraction of 45-50% and a prosthetic aortic valve with significantly increased transvalvular gradients (peak gradient of 68 and mean gradient of 43 mmHg). Transesophageal echocardiogram revealed restricted motion of one of the leaflets without obvious vegetations or masses. Fluoroscopy in multiple views confirmed prosthetic valve dysfunction with only one functioning leaflet.

Decision-making:
Given the sudden onset of symptoms, thrombosis was suspected as the most likely etiology. The patient opted against re-do surgery, thus a decision was made to attempt thrombolysis.

After several hours of an unfractionated heparin (UH) drip, thrombolysis with Alteplase was attempted with a 10mg IV bolus followed by a 90mg infusion. The infusion was breviated due to severe abdominal pain and concern for bleeding, but her hemodynamics and hematocrit remained stable. The efficacy of the therapy was assessed by repeat TTE and fluoroscopy. TTE showed a peak gradient of 37.5 and a mean gradient of 17mmHg (within the normal range for this valve type). Fluoroscopy showed good motion of both valve leaflets. She showed significant clinical improvement. UH infusion was continued as a bridge to a new higher INR goal of 3.5 (3.0-4.0).

Conclusion:
This case demonstrates the importance of including valve thrombosis in the differential diagnosis of valve malfunction even in the setting of uninterrupted anticoagulation with goal INR. It also stresses good history taking and building a contextualized, patient-relevant differential diagnosis.
Abstract 343: Vascular Inflammation in Two Rat Models of Arterial Hypertension is Promoted by Coagulation FXI
Authors: Lagrange Jeremy , Kossmann Sabine , Daiber Andreas , Oelze Matthias , Monia Brett , Ruf Wolfram , Münzel Thomas , Wenzel Philip
Published: June 2021

Backgroud:
Interactions of platelets, leukocytes and the vessel wall play pivotal roles in activating coagulation and precipitating thrombosis. We were recently able to uncover an angiotensin II (ATII) driven factor XI (FXI)-thrombin amplification loop leading to vascular injury in experimental hypertension in mice.

Objective:
We wanted was to explore the role of thrombin-FXI feedback loop in different models of arterial hypertension in rats.

Methods:
ATII treated wistar rats (1mg·kg
-1
·d
-1
for 7 days using osmotic minipumps) and 5/6 nephrectomized were used for this study. During 2 weeks rats were treated with a FXI antisense oligonucleotide (ASO) (1 week after nephrectomy or 2 weeks before ATII pump implantation, respectively). Blood pressure was recorded with tail cuff measurement. Fluorescence oxidative microtopography was used to evaluate vascular ROS production. Vascular reactivity was assessed in isolated aortic segment. Calibrated automated thrombography was used to measure thrombin generation.

Results:
In ATII infused rats as well as 5/6 nephrectomized rats vascular dysfunction related to hypertension was attenuated when rats were treated with FXI ASO. Hypertension induced VCAM-1 expression was normalize with inhibition of FXI. ROS formation was normalized in ATII infused rats as well as 5/6 nephrectomized treated with FXI ASO. Thrombin generation in platelet rich plasma from 5/6 nephrectomized rats was completely abolished when FXI was inhibited. Finally the overall blood pressure increase was abrogated by FXI ASO treatment in 5/6 nephrectomized rats.

Conclusion:
FXI plays a critical role in a FXI-thrombin feedback loop in hypertension. This pathway is relevant in mice and rats and we were able to very recently obtain the first conclusive results in humans. FXI could be a novel therapeutic target to interrupt this heterotypic cellular coagulation-inflammatory circuit.
Abstract 348: Loss of Cadherin-11 Prevents Aortic Valve Interstitial Cell Contraction
Authors: Bowler Meghan , Merryman W. David
Published: June 2021

Introduction:
Calcific aortic valve disease affects 25% of the Western population over 65. Dystrophic calcification is initiated by TGF-β-mediated myofibroblastic differentiation of resident aortic valve interstitial cells (AVICs), which exhibit increased cell contractility and a subsequent imbalance of intercellular tension, which is speculated to lead to apoptosis of AVICs. Cadherin-11 (CDH11), a mechanosensitive transmembrane protein involved in cell-cell adhesion, is responsible for mediating much of this intercellular tension. While cadherins are known to complex with catenins intracellularly, downstream signaling is poorly understood. Both CDH11 and α-smooth muscle actin (αSMA), a myofibroblast marker involved in cell contractility, are overexpressed in diseased human valves, though previous
in vitro
work suggests they have an inverse relationship.

Materials and Methods:
AVICs were isolated from immortalized WT, CDH11
+/-
and CDH11
-/-
mice. They were exposed to 24 hours of 10% equibiaxial strain on the Flexcell-4000 system to recapitulate the mechanical environment of the valves and analyzed for αSMA, β-catenin, and p120-catenin via western blot. AVICs were also seeded on collagen gels and treated with 1 ng/ml murine TGF-β1 to assay contractility or plated on coverslips and stained for myosin light chain II (MLC II).

Results and Discussion:
Loss of CDH11 in AVICs results in a dramatic increase in αSMA (
A
). Though this suggests the cells would be highly contractile, we observe reduced contractility in CDH11
+/-
and CDH11
-/-
AVICs (
B
). Increases in β-catenin and p120-catenin expression indicate dramatic changes in intracellular cadherin signaling (
A
), and immunofluorescence shows a decrease in MLC II (
C
). Thus, loss of CDH11 may prevent contraction (
B
) by either preventing linkage between contractile AVICs or preventing MLC II expression. These results indicate that CDH11 may be a promising therapeutic target for calcific aortic valve disease.