bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 597: Disruptions in Hepatic Insulin Signaling Reveal an Enterohepatic Signaling Pathway that Regulates the ABCG5 ABCG8 Sterol Transporter and Biliary Cholesterol Secretion
Authors: Ji Ailing , Wang Xuebing , Pijut Sonja , Bennett Lisa , van der Westhuyzen Deneys R , Graf Gregory A
Published: March 2019

Insulin resistance is associated with increased risk for cholesterol gallstones as well as the development of diabetic dyslipidemia. HDL is the primary cholesterol carrier in the reverse cholesterol transport (RCT) pathway, the process by which cholesterol is delivered from peripheral organs to the liver for elimination in bile. Therefore, we hypothesized that insulin signaling regulates hepatobiliary cholesterol transport in the RCT pathway. To test the role of insulin signaling, we utilized mice harboring insulin receptor flanked by loxP sites (IR
fl/fl
) in combination with adenoassociated viral vectors containing no transgene (empty) or Cre recombinase to generate control and liver insulin receptor knock out (LIRKO) mice, respectively. As with previous LIRKO models, our mice showed markedly reduced insulin receptor mRNA and protein in liver, but not skeletal muscle or adipose tissue, and impaired glucose tolerance. LIRKO mice had increased biliary cholesterol secretion as well as increased expression of the ABCG5 ABCG8 sterol transporter, the primary mediator of biliary cholesterol secretion. Levels of SR-BI, the primary HDL receptor, were unchanged as were rates of HDL clearance from plasma and selective delivery of HDL cholesterol to the liver. We also observed increased ileal fibroblast growth factor (FGF)15 mRNA. Wild-type mice treated with FGF19 exhibited increased ABCG5 ABCG8 protein expression in the liver. Immunofluorescence microscopy was used to examine the subcellular localization of ABCG5 in the liver following FGF19 administration. Under control conditions, G5 appeared in puncta, diffusely distributed within hepatocytes. Following treatment with FGF19, G5 signal intensity was substantially increased and juxtaposed to zonula occcludin-1 (ZO-1), a tight junction protein the delineates the canalicular channels within the liver. In conclusion, depletion of hepatic insulin receptors increases G5G8 abundance and promotes its localization to the canalicular surface. This effect is associated with an increase in ileal FGF15 expression which promotes its localization to the apical surface and drives biliary cholesterol secretion.
Abstract 600: Hepatic (Pro)renin Receptor Inhibition Decreases Plasma Cholesterol and Triglyceride Levels but Does Not Ameliorate Atherosclerosis in LDLR
-/-
Mice
Authors: Ren Liwei , Sun Yuan , Danser A.H. Jan , Lu Xifeng
Published: March 2019

Rationale:
Cardiovascular diseases (CVD) are the leading cause of death worldwide. Atherosclerosis, dyslipidemia, hypertension, obesity and diabetes are major risk factors for CVD. Elevated plasma cholesterol, especially low-density lipoprotein cholesterol (LDL-c), accelerates atherosclerosis development by inducing lipid accumulation in the vascular wall, foam cell formation and chronic inflammation. Hypertriglyceridemia has also been identified as a risk factor for CVD. Recently, we have identified the (pro)renin receptor [(P)RR], a signaling receptor for renin/prorenin, as a key regulator of lipid metabolism. Inhibiting the (P)RR receptor prevented diet-induced obesity and dyslipidemia in C57BL/6J mice, suggesting that (P)RR inhibition might be used to treat atherosclerosis. Therefore, in the current study, we aimed to explore whether inhibiting the hepatic (P)RR in LDLR
-/-
mice could reduce plasma lipid levels and attenuate atherosclerosis.

Methods and Results:
Eight week-old male LDLR
-/-
mice were injected with GalNAc-modified (liver-specific) antisense oligos against the (P)RR weekly, and fed a high-fat diet (HFD) for 16 weeks. Inhibiting the hepatic (P)RR prevented HFD-induced obesity in LDLR
-/-
mice, similar to our previous observation in C57BL/6J mice. Moreover, (P)RR inhibition decreased the HFD-induced increase in plasma cholesterol levels by almost 50%. FPLC analysis revealed that the VLDL fraction but not the IDL/LDL fraction was reduced by (P)RR inhibition. Plasma triglyceride levels decreased from 1249 ± 136 to 207 ± 32 mg/dL. Remarkably, despite these changes, (P)RR inhibition did not reduce atherosclerotic lesion areas in the aortic root, and even increased plaque size in the entire aorta.

Conclusion:
Hepatic (P)RR inhibition reduces obesity and plasma cholesterol in HFD-fed LDLR
-/-
mice, yet failed to ameliorate atherosclerosis. The latter most likely relates to the lack of effect of (P)RR inhibition on LDL-c, and implies that only LDL-c, and not triglycerides contribute to atherosclerosis progression.
Abstract 601: MicroRNAs and Hedgehog Signaling in the Pathogenesis of Progressive Liver Injury from NAFLD to Fibrosis
Authors: Cheng Xiao , Harris Edward , Cui Juan , Su Neetu
Published: March 2019

Non-alcoholic fatty liver disease (NAFLD) is very common (~70%) in subjects with diabetes and is associated, independently of several confounding factors, with an increased risk of cardiovascular disease (CVD). Diabetes is an independent risk factor for progression of liver fibrosis. Recent studies in human subjects with NAFLD and animal models have demonstrated that microRNAs, a group of noncoding small RNAs, are intimately involved in the development and progression of liver injury, and act to alter expression of genes involved in lipid metabolism and apoptosis. MiR-34a and miR-29 family were among the most frequently dysregulated miRNAs in NAFLD. Thus, the objective of this study was to determine the role of miR-34a and miR-29 in the onset and progression of hepatic steatosis to liver fibrosis in a diabetic mouse model. We showed that streptozotocin (STZ)-induced diabetes predisposed mice to a high-fat-diet (HFD) induced severe liver injury in a much short time course compared to non-diabetic mice. The livers exhibited the development of a spectrum pathological change, with hepatic steatosis at week 6, NASH at week 8 and liver fibrosis at week 12 upon high fat feeding. More importantly, the progressive live injury was closely associated with significantly increased or reduced expression of miR-34a and miR-29 family, respectively, resulting in the activation of hedgehog signaling and the markers of fibrogenesis, Col1A1 and alpha-SMA. Treatment with nanoparticles carrying miR-29b1 mimics improved insulin sensitivity and prevented HFD induced liver fibrosis. In vitro, treatment of McA-RH7777 cells, a rat hepatoma cell line, with a hedgehog signaling inhibitor MDB5 prevented overexpression of miR-34a induced by a free fatty acid, palmitic acid but significantly enhanced expression of miR-29 family in both hepatocytes and hepatic stellate cells. This action hindered the activation of fibrogenesis by lipotoxicity and inflammatory cytokine TNFα. Our study, for the first time, delineates the interaction between miR-34a and miR-29 family and hedgehog signaling in the development of NAFLD and liver fibrosis. This novel finding may provide rationale for developing miRNAs as pharmaceutical targets for the prevention and treatment of liver injury.
Abstract 603: Sequences Proximate to the Renin Cleavage Site in Angiotensinogen Do Not Affect Angiotensin II-mediated Functions
Authors: Wu Chia-Hua , Wu Congqing , Ye Feiming , Howatt Deborah , Balakrishnan Anju , Moorleghen Jessica , Vander Kooi Craig , Daugherty Alan , Lu Hong
Published: March 2019

Objective:
Angiotensinogen (AGT) cleavage by renin is the rate limiting step to produce angiotensin (Ang)II, a critical contributor to hypertension and atherosclerosis. Human AGT can not be cleaved by mouse renin as demonstrated by a transgenic mouse model expressing human AGT. Amino acids at positions 11 and 12, adjacent to the renin cleavage site in AGT, have been proposed to regulate renin cleavage between human and mouse. This study determined whether these two residues affect renin cleavage and the consequent AngII-mediated functions.

Methods and Results:
Hepatocyte-specific AGT deficient (hepAGT-/-) mice have low plasma AGT but high renin concentrations. This mouse model injected with adeno-associated viral vectors (AAV) encoding AGT was used to repopulate AGT. We first determined whether repopulation of human AGT using AAV recapitulate phenotypes of human AGT transgenic mice. HepAGT-/- mice were injected with AAV having a null insert or encoding human AGT, while wild type littermates (hepAGT+/+) were injected with the null AAV. Administration of AAV encoding human AGT led to high plasma human AGT concentrations without affecting plasma mouse renin concentrations, and had no effect on blood pressure and atherosclerosis. In a subsequent study, AAV encoding mutated mouse AGT with Leu11Val and Tyr12Ile that were same as the two residues in human were injected into hepAGT-/- mice. Repopulation of the mutated mouse AGT resulted in increased plasma mouse AGT concentrations and reduced renin concentrations that were comparable to their concentrations in hepAGT+/+ mice injected with the null AAV. Consequently, AAV-driven expression of mutated mouse AGT increased blood pressure and atherosclerosis in hepAGT-/- mice that were comparable to their magnitudes in hepAGT+/+ mice injected with the null AAV.

Conclusion:
The two amino acids proximate to renin cleavage site do not affect AngII-mediated effects.
Abstract 609: Pro-atherosclerotic Low-density Neutrophils Are Present in Hypercholesterolemia
Authors: Cochran Blake J , Pinheiro Damilola , Prendecki Maria , Jones Ben , Cegla Jaimini , Lee Victoria , Barter Philip J , Murphy Andrew J , Rye Kerry-Anne , Woollard Kevin J
Published: March 2019

Introduction:
Elevated cholesterol levels and increased accumulation of myeloid cells are key risk factors in the development of atherosclerosis. Whilst there is significant understanding of the impact of hypercholesterolemia on monocyte function, less is known about the impact of cholesterol on neutrophils. Neutrophils contain the necessary machinery to take up, synthesize, efflux and esterify cholesterol and unique neutrophil phenotypes have been shown in various pathologies, including low density neutrophils (LDNs) described in inflammatory diseases.

Aim:
Determine if cholesterol loading of neutrophils occurs in hypercholesterolemia and characterize the associated phenotypic and functional consequences.

Methods:
Patients (n=13) were a mix of familial hypercholesterolemia and mixed dyslipidemia all with total cholesterol > 7.0 mmol/L. Age and sex matched healthy donors were used as controls. Normal and low density neutrophils were isolated using a percoll density protocol and further purified by FACS selecting for CD66b
+
CD15
+
cells.

Results:
LDNs were only present in hypercholesterolemic patients, accounting for 12.29.6% of the cells in the lower density percoll fraction (p<0.01 vs control). LDNs were morphologically similar to mature high density neutrophils and stained positive for neutral lipid. Functionally, LDNs secreted significantly more extracellular traps (NETs) in response to PMA treatment relative to normal density neutrophils isolated from the same patients or healthy controls (p<0.005). Cholesterol loaded neutrophils were more adhesive to endothelial cells compared to controls (p<0.005). Increased NETosis and neutrophil adhesion have been demonstrated to be atherogenic. Phagocytosis was increased in LDNs vs controls (p<0.05). These functional changes could all be replicated in vitro via treatment of neutrophils from healthy subjects with cholesterol or oxLDL.

Conclusion:
Hypercholesterolemia results in cholesterol loading and a pro-atherosclerotic phenotype in neutrophils. Our data provides evidence that this cholesterol loading occurs in the circulation. More work is now being carried out to further characterize the phenotype of LDNs and mechanisms of maturation during hypercholesterolemia.
Abstract 613: Disturbed Flow-induced Telomeric Repeat Binding Factor 2 (terf2)-interacting Protein (terf2ip) K240 Sumoylation is Critical for Endothelial Senescence but Not Inflammation
Authors: Fujii Yuka , Le Nhat-Tu , Heo Kyung-Sun , Vu Hang T , Kolta Sivareddy , Ko Kyung-Ae , Wang Yin , Thomas Tamlyn N , McBeath Elena , Giancursio Carolyn , Fujiwara Keigi , Abe Jun-ichi
Published: March 2019

Upregulation of both inflammation and senescence (Sen) of endothelial cells (ECs) in the disturbed flow area within the aortic arch is well established, but the exact mechanisms and the relationship between inflammation and senescence remain unclear. TERF2IP is involved in regulation of DNA double strand break (DSB) repair of telomeres (TLs) as a component of the shelterin complex and in IκB kinase (IKK)-NF-kB signaling in the cytosol. TERF2IP and TERF2 form a stable heterodimer to protect the duplex region of TLs. Recent studies showed that TERF2IP or TERF2 deficiency caused TL shortening in mice. We studied how TERF2IP SUMOylation is regulated in ECs and what role it may play in EC inflammation and Sen. We found that disturbed flow (d-flow) activated p90RSK and increased TERF2IP S205 phosphorylation and subsequently K240 SUMOylation. Overexpression of dominant negative p90RSK inhibited d-flow-induced TERF2IP S205 phosphorylation and K240 SUMOylation, and also inhibited EC inflammation and Sen provoked by d-flow. TERF2IP S205 phosphorylation induced TERF2IP nuclear export, which then caused EC inflammation and Sen. In contrast, TERF2IP K240 SUMOylation had no effect on TERF2IP nuclear export, and the TERF2IP K240R SUMOylation site mutant failed to inhibit d-flow-induced NF-kB activation. Interestingly, this point mutation inhibited d-flow-induced Sen with reduced d-flow-induced TL shortening, 8-OHdG level, and apoptosis. Overexpression of a de-SUMOylation enzyme SENP2 and SENP2 T368 mutant significantly inhibited d-flow-induced TERF2IP SUMOylation, suggesting that SENP2 regulates nuclear TERF2IP SUMOylation. Lastly, EC Sen increased when SENP2 was specifically knocked out in ECs in mice, suggesting that TERF2IP K240 SUMOylation may play a role in d-flow-induced EC Sen. These data suggest that TERF2IP post-translational modifications play roles in regulating EC inflammation and Sen, which are differentially regulated by S205 phosphorylation and K240 SUMOylation. TERF2IP K240R knock-in mice will be used to determine the role of TERF2IP K240 SUMOylation in d-flow-induced atherogenesis. We suggest that TERF2IP SUMOylation promotes TL dysfunction, but it does not interfere with TERF2IP-mediated IKK-NF-kB activation in the cytosol.
Abstract 617: Apolipoprotein E Receptor-2 Deficiency Accelerates Smooth Muscle Cell Senescence via Impairment of Protein Phosphatase-2-mediated Cytokinesis
Authors: Komaravolu Ravi K , Waltmann Meaghan D , Jaeschke Anja , Hui David Y
Published: March 2019

Objective:
Genome-wide association studies revealed apoE receptor-2 (apoER2) variants increase atherosclerosis risk in humans. ApoER2 deficiency in hypercholesterolemic

Ldlr
-/-

mice also displayed accelerated atherosclerosis with more complex lesions and extensive lesion necrosis. While loss of apoER2 in macrophages contributes to the advanced lesion phenotype, apoER2 is also expressed in vascular smooth muscle cells (SMC). This study explores the role of apoER2 in SMC functions.

Methods and Results:
Freshly isolated aortic SMC from apoER2 knockout mice showed ~40% less proliferative capacity and displayed a significant reduction in their population doubling when compared to SMC from wild-type (WT) mice. Despite the reduced proliferative rate, apoER2-deficient SMC responded more robustly to TGF-β induced fibronectin synthesis, suggesting a senescence-associated secretory phenotype with increased fibrosis. Accelerated senescence of apoER2-deficient SMC was confirmed by cytochemical staining of β-galactosidase activity and by the increased number of multinucleated polyploidy cells compared to control. Western blot analysis of cell cycle proteins revealed the accumulation of cyclin B1, D1 and securin in apoER2-deficient SMC, indicating that loss of apoER2 does not affect cell cycle entry but promotes cell cycle arrest at the metaphase/anaphase. Increased levels of phosphorylated aurora kinases A/B/C were also observed in apoER2-deficient SMC, suggesting the impairment of protein phosphatases during cell cycle exit. Interestingly, co-immunoprecipitation experiments showed that apoER2 interacts with the catalytic subunit of protein phosphatase 2A (PP2A-C). In the absence of apoER2, PP2A-C failed to interact with cell-division cycle protein 20 (CDC20) thus resulting in inactive anaphase promoting complex (APC/C
CDC20
) and the impairment of cell cycle exit.

Conclusions:
This study identifies a novel function of the lipoprotein receptor apoER2 in SMC mitosis. The data indicate that apoER2 promotes the recruitment of PP2A-C to the anaphase complex, and the impairment of this process accelerates premature smooth muscle cell senescence similar to that observed in the aging vessel wall.
Abstract 627: Combined Plaque Evaluation by Ultrasound and Microarrays Reveals Bclaf1 as a Novel Regulator of Smooth Muscle Cell Transdifferentiation in Atherosclerosis
Authors: Rykaczewska Urszula , Saliba-Gustafsson Peter , Lengquist Mariette , Kronqvist Malin , Lund Kent , Caidahl Kenneth , Skogsberg Josefin , Vukojevic Vladana , Lindeman Jan H , Paulsson-Berne Gabrielle , Hansson Göran K , Leeper Nicholas , Ehrenborg Ewa , Razuvaev Anton , Hedin Ulf , Perisic Matic Ljubica
Published: March 2019

Objective:
Understanding molecular processes behind carotid plaque instability is necessary to develop methods that can identify patients and lesions at risk of stroke. Here, we investigated molecular signatures in human plaques stratified by echogenicity as assessed by duplex ultrasound (US).

Results:
Plaque echogenicity measured by US was correlated to microarray profiles from lesions retrieved at surgery (n=96). Pathway analyses highlighted enrichment of cell apoptosis and proliferation, and BCLAF1 (BCL2 associated factor 1) as the most significantly dysregulated gene (adjusted p<0.0001). BCLAF1 was strongly downregulated in plaques vs. control tissues, positively correlated to markers of cell proliferation and negatively to apoptosis, at both transcriptomic and proteomic level. Immunohistochemistry showed that BCLAF1 was localized in smooth muscle cells (SMCs) nuclei and repressed early during atherogenesis, but reappeared in CD68+ cells in advanced plaques. Proximity ligation assay demonstrated interaction of BCLAF1 with previously reported interaction partners THRAP3 and BCL2, in normal arteries and plaques.
In vitro
, stimulation of SMCs with pro-survival factors EGF, bFGF, PDGFB resulted in induction of BCLAF1, while it was suppressed by macrophage-conditioned medium. Moreover, BCLAF1 silencing in SMCs led to downregulation of BCL2 and SMC markers, and a decrease in proliferation and adhesion (p<0.0001). Transdifferentiation of SMCs using oxLDL, confirmed by CD68 upregulation and MYH11 repression, was accompanied by upregulation of BCLAF1. However, a combination of oxLDL exposure and BCLAF1 silencing, resulted in preserved expression of MYH11 and prevented transdifferentiation. Finally, BCLAF1 expression in CD68+/BCL2+ cells of SMC origin, was verified in plaques from MYH11-lineage tracing atherosclerotic mice.

Conclusions:
Carotid plaque echogenicity correlated with enrichment of molecular pathways associated with cell survival and apoptosis and identified BCLAF1, previously not described in atherosclerosis, as the most dysregulated gene. Functionally, BCLAF1 appeared to promote SMC survival by transdifferentiation into macrophage-like phenotype, by interacting with BCL2 and THRAP3.
Abstract 646: A Simple Multi-Risk-Factor Decision-Making Strategy for Improved Coronary Plaque Burden Increase Prediction: a Patient-Specific 3D FSI Study Using IVUS Follow-up
Authors: Tang Dalin , Wang Liang , Maehara Akiko , Molony David , Samady Habib , Wu Zheyang , Zheng Jie , Mintz Gary S , Giddons Don P
Published: March 2019

Plaque progression and vulnerability are influenced by many risk factors. Our goal is to find simple methods to combine multiple risk factors for better plaque development predictions.

A sample size of 374 intravascular ultrasound (IVUS) slices with matched follow-up was obtained from 9 patients (Mean age 59, 7 m) with informed consent obtained. 3D fluid-structure interaction models were constructed to obtain plaque stress/strain conditions. Four morphological and biomechanical factors (plaque burden (PB), cap thickness (CT), lipid percent (LP) and average plaque wall stress (PWS)) were chosen to predict plaque burden increase defined as

PBI = (PB at follow-up) - (PB at baseline).

For a given slice Si, the ground truth Y
PBI
is define as

Y
PBI
(Si)=1 if PBI(Si)>0; Y
PBI
=0 if PBI(Si)≤0.

For a single predictor W, a threshold value Wc was used to assign the binary prediction outcome:

Y
W
(Si)=1 if W>Wc; Y
W
(Si)=0 if W≤Wc.

Wc was chosen to get optimal agreement between Y
W
and Y
PBI
. To use multiple predictors (say, W1, W2, W3) to PBI, a new predictor Combo(W1,W2,W3) was created with its values defined as

Combo(W1,W2,W3)=Y
W1
+Y
W2
+Y
W3
, where Y
W1
, Y
W2
and Y
W3
were evaluated the same way as before. Combo was then treated as a single predictor and a threshold value was determined to achieve best agreement with Y
PBI
.

Table 1 summarizes the optimal thresholds and agreement rates for all 15 strategies. Agreement rate using PB alone was 57.5%. PWS was the best single predictor for PBI with agreement rate 62.6%. Combining CT and PWS achieved 66.5% agreement rate, 9% better over PB, which was also obtained by combining 4 risk factors.

The method presented here could be used to combine predictors from different sources (stenosis, cap, lipid, inflammation, macrophage, hemorrhage, stress, strain, flow shear stress, FFR, smoking, diabetes, cholesterol, alcohol, hypertension, pro-rupture genes, etc.) to improve prediction accuracy and help decision-making in clinical practice.
Abstract 665: Perivascular Adipose Tissue near Aortic Arch is a Major Site for Atheroprotective IgM Producing B-1 Cells
Authors: Srikakulapu Prasad , Upadhye Aditi , Davy John , McNamara Coleen A
Published: March 2019

Background:
Perivascular adipose tissue (PVAT) regulates artery physiology and pathology, such as promoting atherosclerosis development through local production of inflammatory cytokines. The phenotype of PVAT is region-specific and PVAT composed of brown adipose tissue (near the aortic arch and thoracic aorta) and white adipose tissue (near the abdominal aorta). B-1 cells limit adipose tissue inflammation and atherosclerosis through the production of IgM antibodies. PVAT harbors high numbers of B cells. Id3 is a basic helix-loop-helix protein and important for B cell development. B cell-specific Id3 deficiency (Id3
BKO
) increases B-1b cell numbers and provides atheroprotection. However, the location and regulation of IgM producing B-1 cells in the PVAT are unknown.

Methods and Results:
Flow cytometry analysis of PVAT of normal chow diet fed young ApoE
-/-
mice (n=5) demonstrated that the abundant CD19
+
B cells (per gram fat) harbored in PVAT around aortic arch (2.7
+
0.69 x10
5
cells) compared to PVAT near the thoracic (0.9
+
0.29 x10
5
cells) and abdominal aorta (1.1
+
0.51 x10
5
cells). Interestingly, a large proportion (40%) of these B cells are belong to the CD19
hi
B220
low
B-1 subset in PVAT near aortic arch. CXCL13 is a chemokine, which is important for B-1 cell recruitment to omental fat. Real time-PCR data confirmed that high numbers of B-1 cell recruitment to PVAT near aortic arch is due to high expression of CXCL13 in the PVAT near the aortic arch compared to PVAT near thoracic aorta and abdominal aorta. Moreover, Flow cytometry and ELISPOT data in normal chow fed young ApoE
-/-
mice with Id3
BKO
demonstrated that Id3
BKO
increased B-1b cells (Id3
BKO
: 1.2
+
0.16 x10
3
; Id3
WT
: 0.4
+
0.06 x10
3
; p-value: <0.01; n=6 mice/group) and IgM secreting cells (Id3
BKO
: 2.0
+
0.42 x10
3
; Id3
WT
: 0.78
+
0.23 x10
3
; p-value: <0.05; n=6 mice/group) respectively in PVAT near aortic arch compared to Id3
WT
control mice.

Conclusion:
Results provide the first evidence that atheroprotective B-1 cell profile in PVAT is region-specific and identify Id3 and CXCL13 as regulators of aortic arch PVAT B-1b cell numbers and IgM production.
Abstract 685: Sterol Regulatory Element-Binding Protein 2-Mediated Endothelial-to-Mesenchymal Transition Contributes to Pulmonary Fibrosis
Authors: Martin Marcy , Zhang Jiao , Miao Yifei , Kang Jian , Huang Hsi-Yuan , He Ming , Dong Jianjie , Wong Simon , Huang Hsien-Da , Chen Zhen , Hagood James S , Shyy John Y
Published: March 2019

Objective:
Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease with a median survival rate of 3-5 years. The clinical presentation of IPF relies on the formation of fibroblastic foci, areas of active myofibroblasts depositing extracellular matrix resulting in stiff and dilated alveoli. Whereas therapeutics targeting fibroblast proliferation have made little clinical progress, the role of endothelial cell (EC) dysfunction in pulmonary fibrosis has been overlooked. Having previously identified that sterol regulatory element-binding protein 2 (SREBP2) is a key mediator of innate immune and redox responses in ECs, we postulate that SREBP2 plays a role in promoting EC dysfunction contributing to pulmonary fibrosis.

Approach and Results:
RNA-sequencing demonstrated that SREBP2 overexpression in ECs led to the induction of TGF, Wnt, and cytoskeleton remodeling gene ontology pathways. SREBP2 regulation of these pathways indicated mesenchymal cell differentiation, which alluded to the promotion of endothelial-to-mesenchymal transition (EndoMT). SREBP2 overexpression in cultured ECs led to increased expression of mesenchymal genes such as snai1 family transcriptional repressor 1 (snai1), α-smooth muscle actin (αSMA), vimentin, and neural cadherin (N-cad). Furthermore, SREBP2 was found to directly bind to the promoter regions and transactivate these mesenchymal genes in ECs. Furthermore, a myofibroblast-like phenotypic switch in ECs was evident, including increased proliferation, stress fiber formation, and ECM deposition. Pulmonary fibrosis mouse models using bleomycin demonstrated exacerbated lung vascular remodeling and increased EndoMT in endothelial-specific SREBP2 overexpression transgenic (EC-SREBP2-Tg) mice when compared to wildtype littermates. Additionally, SREBP2 was found to be markedly increased in lung specimens from IPF patients.

Conclusions:
SREBP2 promotes EndoMT through direct transactivation of mesenchymal genes (i.e., snai1, αSMA, vimentin, and N-Cad). Pathogenic factors activating SREBP2 specifically in ECs increased EndoMT and exacerbated pulmonary fibrosis
in vivo
. This suggests that targeting SREBP2 may be a potential therapeutic in pulmonary fibrosis.
Abstract 692: Development of the Gene Therapy With CRE Decoy ODN to Prevent Vascular Intimal Hyperplasia
Authors: Uchida Daiki , Saito Yukihiro , Hirata Satoshi , Azuma Nobuyoshi
Published: March 2019

Background:
Intimal hyperplasia (IH) is the main cause of vein graft stenosis or failure after bypass surgery. However, in the previous study, no therapeutic targets for the treatment of IH have been identified. Our recent research have been reported that the inhibition of Cyclic adenosine monophosphate response-element (CRE) binding protein (CREB) activation is a key role for vein graft IH.

Objective:
In consideration of future clinical application, we focused on decoy oligodeoxynucleotide (ODN) transfection as gene therapy strategy for suppressing IH. The goal of the present study is to identify whether the CRE decoy ODN had the therapeutic efficacy for suppressing IH.

Methods:
We developed phosphorothioate CRE decoy ODN and checked binding ability to a CRE sequence using CREB transcription assay . We chose a decoy ODN having high first-binding ability and transfected it to vascular smooth muscle cell(VSMC) in vitro. Proliferation was assessed using MTS assay and migration was assessed using a modified Boyden-chamber assay. We examined CRE activity using Luciferase reporter gene assay. We checked expression of mRNAs using qRT-PCR. In wire-injury mouse model(C57BL6,n=6), CRE decoy ODN was transfected to injured vessel wall using ultrasound-sonoporation method in vivo.

Results:
CREB transcription factor assay showed CRE decoy had about 80 folds binding ability of control decoy in IC50 value (vs.control,n=5). Transfer of CRE decoy ODN resulted in significantly downregulated CRE activity(vs.control,P<.01,n=12) and inhibited VSMC proliferation (vs.control,P<.01,n=12) and migration (vs.control,P<.01,n=5). In a wire-injury mouse model, histopathological analysis revealed that transfer of the CRE decoy ODN significantly repressed IH whereas intensive IH was observed in the control group. In this vessel tissue, Levels of mRNA CCNA2 and Bcl2 were significantly lower in CRE decoy group than in control group(n=6,P<.01,respectively). CRE-activated gene expression was strongly repressed.

Conclusions:
The present result suggested that CRE decoy ODN inhibiting CREB function provide an effective therapeutic approach to suppressing IH.
Abstract 696: Transgelin in Aortic Abdominal Aneurysm and its Potential Link in Electronic Cigarette Vapor Mediated Cardiovascular Disease
Authors: Ghosh Abhijit , Krause Will , Kasten Seth , Abdallah Andrew , Hirve Anirudh , Pechota Angela , Durham Laura , Dunivant Veronica , Eliason Jonathan L
Published: March 2019

Introduction:
The E-cigarette form of smoking (e-cig) has rapidly increased. Potential adverse cardiovascular effects are unknown. We sought to determine if e-cig use affects Transgelin (TAGLN), an actin binding protein with multiple important cell-signaling and regulatory actions noted to be targeted by autoantibodies in AAA.

Methods:
Human aortas and serum from abdominal aortic aneurysm (AAA) (n=26) and non-AAA (n=15), and rat aortas from elastase-perfused (EP) or saline-perfused (SP) animals (n=3 each group) were analyzed for aortic transgelin (TAGLN) and alpha smooth muscle actin (α-sma) content, serum TAGLN content and/or serum anti-TAGLN antibodies (AB). Rats exposed to e-cig vapor (EV) 90 min/day for 4 or 8 weeks , or room air (RA) (n=6 per group), and rats undergoing aortic low-dose topical elastase (LDTE) to induce inflammation followed by EV or RA x 100 days (n=6 per group), were also analyzed. Aortas were assessed by Western blot for TAGLN and actin content, while serum studies used Elisa to detect circulating TAGLN, or anti-TAGLN antibodies.

Results:
Western blots revealed loss of TAGLN and α-sma in the aortas of AAA compared to control tissues. In rat AAA we observed loss of TAGLN and diminished levels of α-sma in EP compared to SP control aortas. Serum from AAA patients had increased anti-TAGLN antibodies (
P
=0.085, 45% more in AAA) and circulating TAGLN (P=0.008) compared to controls. LDTE EV rat aortas had less TAGLN (
P
=0.002) and α-sma (
P
=0.008) compared to RA rats throughout the aorta. LDTE + EV rat serum had high levels of anti-TAGLN antibodies (P=0.027) and circulating TAGLN (P=0.007) compared to RA rats.

Conclusion:
These results show that AAA is associated with decreased aortic TAGLN in humans and rats in the region of the aneurysm. EV exposure alone, and especially following LDTE results in marked reduction of TAGLN throughout all segments of the aorta. Human AAA patients and EV rats with LDTE also exhibit elevation of circulating TAGLN and anti-TAGLN antibodies. These results suggest an autoimmune mechanism for TAGLN targeting in the aorta when AAA is present or e-cig vapor exposure occurs, and that e-cig use may result in cardiovascular risk.
Abstract 705: Nudt6 Inhibition Limits Smooth Muscle Cell Apoptosis and Abdominal Aortic Aneurysm Progression
Authors: Jin Hong , Winter Hanna , Chernogubova Ekaterina , Li Yuhuang , Winski Greg , Bäcklund Alexandra , Maegdefessel Lars
Published: March 2019

Abdominal Aortic Aneurysm (AAA) is a highly lethal disease, of which diagnosis and treatment is still a major burden. Natural Antisense Transcripts (NAT) are known to inhibit key transcripts by antisense targeting. Their contribution to the development and progression of aneurysm disease has not been studied to date. The aim of this study was to investigate the role of the NAT Nudix Hydrolase 6 (Nudt6) and its antisense target Fibroblast growth factor (Fgf2) in AAA disease, by modulating NUDT6
in vivo
in Angiotensin II (AngII)-induced experimental AAA
via
sonoporation (microbubble procedure) as well as
in vitro
in primary mouse aortic smooth muscle cells (mAoSMCs) via transfection
.
Further, primary mAoSMCs were observed using the IncuCyte imaging system to monitor changes in dynamic processes like proliferation and apoptosis. FGF2 is known to be a potent mitogen of smooth muscle cell (SMC) proliferation; a process being drastically diminished in AAA progression. By inhibiting Nudt6 with GapmeR
in vivo
we could observe smaller aortic diameters, increased both Fgf2 expression and SMCs survival by immunohistochemistry.
In vitro
, proliferation was increased in Nudt6 inhibited AngII-stimulated primary mouse aortic SMCs. Overexpression of Nudt6 did not have a significant effect on proliferation. Nudt6 and Fgf2 gene expression are inversely correlated and experience a switch of expression patterns during AngII stimulation over a longer period of time if primary SMCs are transfected with Nudt6 inhibitor and treated with AngII. All findings suggest a beneficial effect of Nudt6 inhibition in experimental AAA. In the future, it could be used as a potential therapeutic target in AAA by increasing SMC survival rates.
Predicting Future Cardiovascular Events in Patients With Peripheral Artery Disease Using Electronic Health Record Data
Authors: Ross Elsie Gyang , Jung Kenneth , Dudley Joel T. , Li Li , Leeper Nicholas J. , Shah Nigam H.
Published: March 2019

Background:

Patients with peripheral artery disease (PAD) are at risk of major adverse cardiac and cerebrovascular events. There are no readily available risk scores that can accurately identify which patients are most likely to sustain an event, making it difficult to identify those who might benefit from more aggressive intervention. Thus, we aimed to develop a novel predictive model—using machine learning methods on electronic health record data—to identify which PAD patients are most likely to develop major adverse cardiac and cerebrovascular events.

Methods and Results:

Data were derived from patients diagnosed with PAD at 2 tertiary care institutions. Predictive models were built using a common data model that allowed for utilization of both structured (coded) and unstructured (text) data. Only data from time of entry into the health system up to PAD diagnosis were used for modeling. Models were developed and tested using nested cross-validation. A total of 7686 patients were included in learning our predictive models. Utilizing almost 1000 variables, our best predictive model accurately determined which PAD patients would go on to develop major adverse cardiac and cerebrovascular events with an area under the curve of 0.81 (95% CI, 0.80–0.83).

Conclusions:

Machine learning algorithms applied to data in the electronic health record can learn models that accurately identify PAD patients at risk of future major adverse cardiac and cerebrovascular events, highlighting the great potential of electronic health records to provide automated risk stratification for cardiovascular diseases. Common data models that can enable cross-institution research and technology development could potentially be an important aspect of widespread adoption of newer risk-stratification models.
Prognostic Importance of Defibrillator‐Appropriate Shocks and Antitachycardia Pacing in Patients With Mild Heart Failure
Authors: Biton Yitschak , Daimee Usama A. , Baman Jayson R. , Kutyifa Valentina , McNitt Scott , Polonsky Bronislava , Zareba Wojciech , Goldenberg Ilan
Published: March 2019

Background

Patients with heart failure and an implantable cardioverter‐defibrillator (
ICD
) for primary prevention are at increased mortality risk after receiving shock therapy. We sought to determine the prognostic significance of
ICD
therapies, both shock and antitachycardia pacing, delivered for different ventricular arrhythmia (
VA
) rates.

Methods and Results

We evaluated mortality risk among 1790
ICD
‐implanted patients from
MADIT
‐CRT (Multicenter Automatic Defibrillator Implantation Trial With Cardiac Resynchronization Therapy). For the first analysis, patients were divided into mutually exclusive groups by the rate of treated
VA
only: slow
VA
(<200 beats per minute) and fast
VA
(≥200 beats per minute or ventricular fibrillation). In a secondary analysis, both the type of
ICD
therapy and
VA
rate were used. The reference group was always patients who had no
ICD
therapy.
ICD
therapy for fast
VA
was associated with increased mortality risk (hazard ratio [
HR]
, 2.27; 95%
CI
, 1.48–3.48;
P
<0.001). However, mortality risk after
ICD
therapy for slow
VA
was similar to the risk related to no
ICD
therapy (
HR
, 1.45; 95%
CI
, 0.86–2.44;
P
=0.162). Consistently, shocks (
HR
, 2.96; 95%
CI
, 1.91–4.60;
P
<0.001) and antitachycardia pacing (
HR
, 2.22; 95%
CI
, 0.96–5.14;
P
=0.063) for fast
VA
were both associated with increased mortality risk. Shocks and antitachycardia pacing for slow
VA
were not significantly associated with increased mortality risk (
HR
, 1.43 [95%
CI
, 0.52–3.92;
P
=0.489]; and
HR
, 1.43 [95%
CI,
0.80–2.56;
P
=0.232], respectively).

Conclusions

In patients with mild heart failure receiving
ICD
for primary prevention, mortality is associated with the rate of underlying
VA
rather than the type of therapy. These findings suggest that fast
VA
is a marker for increased mortality rather than shock therapy directly contributing to increased risk.

Clinical Trial Registration

URL
:
http://www.clinicaltrials.gov
. Unique identifier:
NCT
00180271.
A new species and a new combination of the genus Arisaema (Araceae) from China
Authors: MA ZHENG-XU , DU WEN-YAN , WANG XIAO-YUN
Published: March 2019

A new species, Arisaema melanostomum, and a new combination, A. yunnanense subsp. quinquelobatum, are proposed, described and illustrated in this article.
Taxonomy of European Damaeidae (Acari: Oribatida) IV. Partial revision of Metabelba Grandjean, 1936 with proposal of one new subgenus, one new species and redescriptions of two known species
Authors: MOUREK JAN , MIKO LADISLAV , BERNINI FABIO
Published: March 2019

We review the current state of taxonomy of the oribatid mite genus Metabelba Grandjean, 1936 (Damaeidae) and present an amended generic diagnosis. In contrast to Subías (2004) we consider Allobelba Kunst, 1961 and Neobelba BulanovaZachvatkina, 1967 as separate genera, not subgenera of Metabelba. We consider the subgenus Metabelba (Parametabel- ba) Mihelčič, 1964, to be a nomen nudum and propose a new subgenus—Metabelba (Pateribelba) n. subg., with type species Metabelba (Pateribelba) sphagni Strenzke, 1950—to replace it. Pateribelba n. subg. includes species of Metabelba without the propodolateral apophysis. Together with the type species, the following named species (new combinations) belong to this subgenus: Metabelba (Pateribelba) italica (Sellnick, 1931); M. (P.) romandiolae (Sellnick, 1943); M. (P.) platynotus Grandjean, 1954; M. (P.) ericius Kunst, 1957; M. (P.) rhodopeia Kunst, 1961; M. (P.) filippovi Bulanova-Zachvatkina, 1965; M. (P.) flagelliseta Bulanova-Zachvatkina, 1965; M. (P.) monilipeda Bulanova-Zachvatkina, 1965; M. (P.) pseudoitalica Bulanova-Zachvatkina, 1965; M. (P.) paraitalica Kulijev, 1967, M. (P.) glabriseta Mahunka, 1982 and M. (P.) machadoi Pérez-Iñigo, 1986. A new species Metabelba (Pateribelba) denscanis n. sp. is described and illustrated; it is characterized mainly by the presence of a notogastral cornicle in the adult. It was found in Central Bohemia (Czech Republic), Upper Silesia (Poland) and South Tyrol (Italy). Metabelba romandiolae and M. sphagni are redescribed and illustrated. Metabelba gladiator Mihelčič, 1963 is considered to be junior synonym of M. romandiolae. Metabelba lanceolata van der Hammen, 1952 is considered to be a junior synonym of M. sphagni. We also provide a ten-tative diagnostic key of the known species of Metabelba (Pateribelba) n. subg., based on detailed study of available species and a review of literature data.
A new sclerite-free genus and species of Clavulariidae (Coelenterata: Octocorallia)
Authors: ALDERSLADE PHILIP , MCFADDEN CATHERINE S.
Published: March 2019

This short communication describing only a single new taxon is necessary to facilitate further research publications by Neil Blackstone & Austin Parrin (Northern Illinois University) and colleagues. Neil and Austin have found the new species to be an ideal experimental animal that is fast-growing and extremely easy to maintain in laboratory aquaria (Parrin et al. 2010). [NTM = Museum & Art Gallery of the Northern Territory, PO Box 4646, Darwin, NT 0801, Australia].
New species of oribatid mites in the families Synichotritiidae and Phthiracaridae from Thailand, with a checklist of Thai Euptyctima (Acari: Oribatida: Euphthiracaroidea, Phthiracaroidea)
Authors: FUANGARWORN MARUT , LEKPRAYOON CHARIYA
Published: March 2019

Two new species of oribatid mites, Temburongia similanensis sp. nov. (Synichotritiidae) and Austrophthiracarus trapezoides sp. nov. (Phthiracaridae), are described based on adult specimens from leaf litter on Similan Islands, Thailand. Temburongia similanensis sp. nov. differs from T. patoi Mahunka, 1990 in having simple prodorsal and notogastral setae, one pair of median prodorsal cristae, two pairs of exobothridial setae, two longitudinal rows of ano-adanal setae, coxal setation of 3-1-2-3, tridactylous legs, and, more unusually, clusters of bothridial brachytrachea which are unknown in other synichotritiid mites. Austrophthiracarus trapezoides sp. nov. can be distinguished from the similar species A. pseudotuberculatus Mahunka, 2008 by the presence of a large trapezoid prodorsal tubercle and longer notogastral setae (seta c 1 reaching d 1 ). A checklist of Thai Euptyctima, including 19 species in four families (Oribotritiidae, Euphthiracaridae, Synichotritiidae, and Phthiracaridae), their synonyms and distributional data are presented. Plonaphacarus semiaciculatus (Mahunka, 2008) comb. nov. is proposed as a new combination for Rhacaplacarus semiaciculatus Mahunka, 2008.
A probably invasive new genus and new species of soft coral (Octocorallia: Alcyonacea: Clavulariidae) from Brazil
Authors: VAN OFWEGEN L. P. , HADDAD M. A.
Published: March 2019

Stragulum bicolor n. gen. & n. sp. (Octocorallia: Clavulariidae) is described from Brazil. It was found in shallow waters of the States of Ceará, Rio de Janeiro, São Paulo, Paraná and Santa Catarina, and is probably invasive. The genus forms encrusting sheets on solid objects, such as seaweed basal stems, barnacle shells and artificial substrates. The new taxon has spindles, crosses and radiates in the outer layer of the coenenchyme, fused sclerites in the interior and the polyps are unarmed. The new genus is compared with other taxa that have fused sclerites.
A review of the genera Elydnodes Hampson, 1913 and Antitrisuloides Holloway, 1985 with description of two new species (Lepidoptera, Noctuidae). Revision of Pantheinae, contribution III
Authors: BEHOUNEK GOTTFRIED , KONONENKO VLADIMIR
Published: March 2019

Two genera in the noctuid subfamily Pantheinae are revised: Elydnodes Hampson, 1913 and Antitrisuloides Holloway, 1985. Two new species (Elydnodes griseata sp. n. and Antitrisuloides siamensis sp. n.) are described from Thailand, Laos and Malaysia.
A new genus and species of anchialine Hymenosomatidae (Crustacea, Decapoda, Brachyura) from Samar, Philippines
Authors: HUSANA DANIEL EDISON M. , TAN SWEE HEE , KASE TOMOKI
Published: March 2019

A new genus and species of brachyuran crab, Samarplax principe (family Hymenosomatidae) is described from an anchialine cave in Samar Island, Philippines. This cavernicolous species lacks rostrum and has degenerated eyes, possesses two small spines at the lateral margin of the carapace, has a proportionally shorter projected merus of the third maxilliped, an almost flat epistome and brush-like setae instead of teeth along the cutting edges of the chelae. The complete loss of visual organs and pigmentation, the long but slender ambulatory legs and large egg size suggest a completely hypogeal lifestyle for this species. This is the first species of Hymenosomatidae recorded from an anchialine cave in the Philippines exhibiting true troglomorphic adaptations.
Sexual morphs and colour variants of Aphis (formerly Toxoptera) odinae (Hemiptera, Aphididae) in Japan
Authors: BLACKMAN ROGER L. , SORIN MASATO , MIYAZAKI MASAHISA
Published: March 2019

Recent molecular studies have shown that the genus Toxoptera is polyphyletic, and in particular that the name of the aphid known since 1952 as Toxoptera odinae (van der Goot) should revert to Aphis odinae (van der Goot), stat. rev. In this paper oviparae, alate males and fundatrices of A. odinae are described for the first time, from Japan. A. odinae is common as a grey-brown or rust-brown aphid on numerous plant species of shrubby habit throughout the Old World tropics and subtropics, but in temperate east Asia much darker forms occur, and Japanese populations of A. odinae include a dark green form not found elsewhere in the world. Multivariate morphometrics were used to confirm that both colour forms of the aphid in Japan were conspecific with samples from other parts of the world. Pergandeida kalopanacis Hori 1927 is a new synonym of A. odinae.
The millipede genus Anaulaciulus Pocock, 1895 in Taiwan, with descriptions of four new species (Diplopoda, Julida, Julidae)
Authors: MIKHALJOVA ELENA V. , GOLOVATCH SERGEI I. , CHANG HSUEH-WEN
Published: March 2019

The genus Anaulaciulus Pocock, 1895 is represented in Taiwan by eight nominate species, all keyed and nearly all mapped, including four new ones: A. immensus sp. nov., A. oligosegmentatus sp. nov., A. multiarticulatus sp. nov. and A. setulifer sp. nov. One species, A. tonginus (Karsch, 1881), is confirmed to be present in the fauna of the island, while the remaining three require revision. Taxonomic and distributional remarks are provided for all of the species.
Systematic revision of Thaumastocoris Kirkaldy (Hemiptera: Heteroptera: Thaumastocoridae)
Authors: NOACK ANN E. , CASSIS GERASIMOS , ROSE HARLEY A.
Published: March 2019

The genus Thaumastocoris is revised. Nine new species are described (T. busso, T. freomooreae, T. kalaako, T. majeri, T. nadeli, T. ohallorani, T. roy, T. safordi, and T. slateri) and the five previously described species are redescribed. A diagnostic key to species is provided, supported with illustrations of key character systems and maps depicting their distributional range. Host plants are tabulated, and biology and host plant associations are discussed.
The correct publication date for mecopteran insects (Choristidae, Bittacidae, and Panorpidae) described by John Obadiah Westwood, resurrecting Panorpa confusa Westwood, 1841, the confused scorpionfly
Authors: SOMMA LOUIS A.
Published: March 2019

I correct the publication dates for one genus and 14 species of Mecoptera (1 choristid, 4 bittacids, and 9 panorpids), and resurrect Panorpa confusa Westwood, 1841. Additionally, I provide a nomenclatural and taxonomic summary for these taxa, and a publication history for all of Westwood’s related publications.