bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 627: Novel Nanoparticles Berberine Chloride Oral Delivery Approaches Targeting PCSK9
Authors: Ochin Chinedu C , Garelnabi Mahdi
Published: June 2021

Introduction:
PCSK9 has been demonstrated to negatively influence the availability of LDL receptors (LDLR) thus predisposing to hypercholesterolemia. The plant alkaloid Berberine has been reported to enhance LDLR function while downregulating the expression of PCSK9 however this compound has limited use due to poor oral bioavailability. The nanotechnology applied to drug delivery helps improve drug pharmacokinetics and biodistribution.

Aims and Objectives:
In this study, we hypothesized that targeted drug delivery through enhanced intestinal M cell absorption would improve the bioavailability of Berberine Chloride and also enhance its pharmacological activity.

Study Design:
This study involved a first phase of optimized Nano-precipitation synthesis and characterization of the RGD functionalized drug delivery PLGA-PEG nanoparticles containing the drug Berberine Chloride along with drug entrapment efficiency and in vitro drug release studies, using Scanning/Transmission Electron Microscopy, Photon correlation Spectroscopy and UV-Spectroscopy. This is followed by a second phase, which consists of the treatment of M Cell cultures produced by co-culturing of Caco2 and Raji B cells using the previously synthesized targeted Nano-medicine. Treated cell culture samples are then analyzed for PCSK9 gene expression and Western blot studies for permeability-glycoprotein (P-gp) and PCSK-9 levels as well as IL1 and associated lipoprotein genes expressions.

Results:
RGD targeted Berberine Chloride encapsulating nanoparticles had a particle size range distribution of 250- 350nm determined through Photon correlation Spectroscopy. Berberine Chloride entrapment efficiency in PLGA-PEG nanoparticles was within the range of 25-50%. This was calculated from the standard curve of Berberine Chloride using UV spectroscopy.

Conclusion:
Favorable drug concentration levels were achieved for Berberine Chloride upon entrapment in RGD targeted PLGA-PEG nanoparticles, providing a promised opportunity that favorable pharmacokinetic and pharmacodynamics results in M cell culture models of Berberine Chloride relationship to PCSK9 levels.
Abstract 634: Silencing miR144 Enhances Regression and Attenutates Progression of Atherosclerosis
Authors: Tarling Elizabeth , Cheng Joan , Cheng Angela , Tontonoz Peter , Sallam Tamer , Vallim Thomas
Published: June 2021

Cholesterol imbalance plays a central role in multiple diseases, particularly cardiovascular disease, the leading cause of death in the United States. Lipid-loaded macrophage foam cells are a critical determining factor in the pathogenesis of atherosclerosis. MicroRNAs are small non-coding RNA molecules that regulate both cholesterol and lipoprotein metabolism, and may significantly contribute to disease progression. We previously identified miR-144 as a novel regulator of lipid metabolism that is also regulated by the nuclear receptor farnesoid X receptor (FXR). We identified the cholesterol transporter ABCA1 as a miR-144 target gene, and showed that increased hepatic expression of miR-144 decreased ABCA1 protein and plasma cholesterol levels. Consequently, we hypothesized that silencing miR-144 may have therapeutic potential because it may increase the levels of atheroprotective HDL particles and enhance reverse cholesterol transport (RCT) due to increased ABCA1 levels in macrophages. Here we present en face analysis of atherosclerosis showing that silencing miR-144 both enhanced regression and attenuated progression of atherosclerosis. In addition, we demonstrate that silencing miR-144 enhances RCT, the process by which macrophage cholesterol is returned to the liver for excretion. Together, these data suggest that this pathway may represent a bona-fide strategy to treat atherosclerosis.
Abstract 640: Genetic Inactivation of Col15a1 in Smooth Muscle Cells Decreases Plaque Size and Decreases Indices of Plaque Stability
Authors: Durgin Brittany G , Cherepanova Olga A , Owens Gary K , Connelly Jessica J
Published: June 2021

In the development of atherosclerosis, it is widely believed that contractile smooth muscle cells (SMC) increase their migratory and proliferative capacity and invade the neointima where they secrete extracellular matrix (ECM) to form a plaque stabilizing fibrous cap. We previously identified that collagen, type XV, alpha 1 (COL15A1) expression is increased in human atherosclerotic plaques and associated with lesion burden. COL15A1 has been shown to link large fibrillar collagens to confer ECM stability. Of interest, siRNA knockdown of
Col15a1
in cultured human aortic SMC results in a decrease in proliferation and an increase in migration of SMC. Taken together, we hypothesize that SMC-derived COL15A1 plays a critical role in atherosclerosis by promoting matrix organization needed for lesion and fibrous cap stability. Due to ambiguity in identifying dedifferentiated SMC without lineage tracing, the contribution of SMC in producing COL15A1 and how loss of SMC-derived COL15A1 alters plaque morphology and cellular composition is unknown. As such, we generated inducible SMC specific lineage tracing (
Myh11
-CreER
T2
ROSA floxed STOP eYFP

ApoE
-/-

)
Col15a1
knockout (SMC-
Col15a1
KO
) and wild-type (SMC-
Col15a1
WT
) mice and placed them on Western diet for 18 weeks. SMC-
Col15a1
KO
exhibited a drastic decrease in overall plaque (~4 fold) and vessel (~1.5 fold) size. In addition, SMC-
Col15a1
KO
have a decrease in overall plaque collagen content (~3 fold) and collagen fiber maturation. With respect to cell type, SMC-
Col15a1
KO
demonstrated a decrease in overall plaque cell number (DAPI+, ~2 fold) and an increase in the proportion of macrophage (LGALS3+) cells and decrease in the proportion of SMC (YFP+) within plaques. SMC-
Col15a1
KO
showed a decrease in lesion SMC proliferative capacity
in vivo
(~2 fold) that was recapitulated
in vitro
in population doubling proliferation experiments using isolated primary aortic SMC from SMC-
Col15a1
WT
and SMC-
Col15a1
KO
. Interestingly, mixing 1:1 of WT and KO SMC lines was able to rescue the proliferation deficit. These studies provide the first evidence that a SMC derived collagen, COL15A1, is a significant and novel player in advanced atherosclerotic plaque progression, stability, and cellular content.
Abstract 643: Type-2 Cannabinoid Receptor Deficiency is Associated with Atherosclerotic Lesion Calcification in Ldlr-null Mice
Authors: Fulmer Makenzie L , Englehaupt Emilee , Garst Chris , Brown Stacy , Thewke Douglas
Published: June 2021

Background:
Calcification of atherosclerotic plaques is associated with vulnerability to rupture and increased risk of myocardial infarction. The mechanism of plaque calcification is unclear, but has been shown to be a cell-mediated process involving complex signaling pathways affecting the osteogenic transcription factor Runt-related transcription factor 2 (Runx2). The type-2 cannabinoid receptor (CB2) modulates processes involved in bone remodeling and our prior studies determined that CB2 alters the composition of early lesions in hyperlipidemic Ldlr
-/-
mice; however, the function of CB2 in plaque calcification is unknown. Therefore, we tested the hypothesis that CB2 modulates plaque calcification by evaluating the effects of systemic CB2 gene deletion on lesion calcification and aortic expression of Runx2 in Ldlr
-/-
mice.

Results:
Groups (n≥8) of 8-week old CB2
+/+
Ldlr
-/-
(WT) and CB2
-/-
Ldlr
-/-
(CB2
-/-
) mice were fed a high fat diet (HFD) for up to 24 weeks. Standard blood plasma analysis showed no difference in HFD-induced hyperlipidemia between WT and CB2
-/-
mice. Aortic levels of endocannabinoids, anandamide and 2-archidonylglycerol, were significantly elevated after 12 weeks of HFD feeding as determined by LC-MS/MS. En face analysis revealed the extent of atherosclerosis in the aortic arch and thoracic aorta did not differ between WT and CB2
-/-
mice, but was ~1.9-fold greater in the abdominal aortas of CB2
-/-
mice (17.0±1.3% vs 9.0±1.3%, p=0.002). Calcification of aortic root lesions was ~2.3 fold greater in CB2
-/-
mice compared to WT mice (12.9±1.1% vs 5.6±1.2%, p=0.002) as revealed by von Kossa staining. Western blot analysis showed significantly increased expression of Runx2 in aortas of WT mice compared to CB2
-/-
after 20 weeks of HFD (2.55±0.25 fold, p<0.05).

Conclusion:
Systemic CB2 deficiency enhances lesion calcification and is associated with altered aortic expression of Runx2. These results provide novel mechanistic insights into the function of CB2 signaling in the pathogenesis of atherosclerosis and vascular calcification that may lead to the development of therapies aimed at stabilizing calcified plaque.
Abstract 658: Unraveling the Controversy of Bisphosphonates as Vascular Calcification Therapy Using a Nanoanalytical Approach
Authors: Ruiz Jessica L , Hutcheson Joshua D , Aikawa Elena
Published: June 2021

Vascular calcification significantly predicts atherosclerotic plaque rupture and cardiovascular events. Retrospective studies of women taking bisphosphonates, a proposed therapy for vascular calcification, paradoxically indicated increased risk in patients with prior acute events. We recently demonstrated that calcifying extracellular vesicles (EVs) released by cells within the plaque aggregate and nucleate calcific mineral, but the underlying mechanism and the potential for pharmacological intervention remain poorly understood. We hypothesize that bisphosphonates block EV aggregation and arrest existing mineral growth, freezing calcifications in a high-risk morphology that hastens plaque rupture. This study visualized for the first time EV aggregation and calcification at single-EV resolution, via scanning electron microscopy. Three-dimensional (3-D) collagen hydrogels incubated with calcifying EVs modeled fibrous cap calcification, serving as an in vitro platform to image mineral nucleation and test candidate drugs for the potential to inhibit or reverse vascular calcification. EVs aggregated along and between collagen fibrils. Energy-dispersive x-ray spectroscopy (EDS) confirmed that EV aggregates contained calcium and phosphorous, the building blocks of calcific mineral (vs. internal collagen control, p<0.001). The addition of the bisphosphonate ibandronate decreased the EDS-detected amount of calcium (4.32% by weight (wt%) vs. 2.36 wt%, p<0.001) and phosphorous (4.26 wt% vs. 1.94 wt%, p<0.001) comprising EV aggregates. Further, ibandronate reduced the size (21.5 μm
2
vs. 14.2 μm
2
, p=0.012) and changed the morphology of calcific EV aggregates (Figure). These findings agree with our hypothesis that bisphosphonates alter EV-driven calcification, and confirm that our 3-D collagen hydrogel system is a viable platform to study EV-mediated mineral nucleation and evaluate potential therapies for cardiovascular calcification.
Abstract 95: Naringenin Supplementation of a Low-Fat Diet Enhances the Reversal of Metabolic Dysfunction, Promotes Atherosclerosis Regression and Improves Lesion Pathology in
Ldlr
-/-
Mice
Authors: Burke Amy C , Sutherland Brian G , Sawyez Cynthia G , Telford Dawn E , Huff Murray W
Published: June 2021

Diet-induced metabolic dysregulation (MetS) and atherosclerosis in mouse models can be reversed by intervention with a low-fat diet. Supplementation of a high-fat diet (HFHC) with naringenin prevents development of abnormal lipid and glucose metabolism and attenuates atherosclerosis. In the present study, we hypothesized that intervention by the addition of naringenin to a low-fat diet would enhance the reversal of MetS and atherosclerosis.

Ldlr
-/-

mice were fed a HFHC diet for 12 weeks to induce MetS and intermediate atherosclerosis (baseline). Intervention for an additional 12 weeks consisted of transfer to: 1) a low-fat, isoflavone-free chow diet (IFF), 2) IFF with 3% naringenin (IFF+Nar) or 3) continuation on HFHC. HFHC-feeding induced rapid weight gain and adiposity, which were reversed to a greater extent by intervention with IFF+Nar (-93% and -76%,
P
<0.05) compared to IFF (-60% and -46%,
P
<0.05), independent of caloric intake. The hypercholesterolemia and hypertriglyceridemia induced by HFHC were further decreased by intervention with IFF+Nar (-105% and -124%,
P
<0.05) compared to IFF (-96% vs -103%,
P
<0.05). Hepatic lipids were normalized by both IFF+Nar and IFF, although IFF+Nar induced a greater decrease in liver triglyceride (-110% vs -86%,
P
=0.05). Hepatic expression of
Pgc1a
,
Cpt1a
and
Pnpla2
were significantly higher with IFF+Nar compared to IFF. Fasting plasma glucose, plasma insulin and insulin sensitivity were further improved by IFF+Nar, compared to IFF. Relative to baseline, aortic cholesteryl ester (CE) increased with intervention by IFF alone (+40%), whereas IFF+Nar reversed aortic CE content (-19%,
P
<0.04). Compared to baseline, aortic sinus lesion size continued to increase with IFF (+47%), whereas with IFF+Nar lesion size increased only 14% (
P
<0.05), indicating almost complete attenuation of lesion growth. Both intervention diets decreased lesion apoptotic cells similarly, although fewer lesion macrophages (35% vs 43%,
P
<0.05) and reduced necrotic area (6.5% vs 7.5%, trend) resulted from intervention with IFF+Nar, compared to IFF. In conclusion, intervention with naringenin enhances improvements in metabolic function, halts progression of atherosclerosis and improves lesion pathology.
Abstract P631: Hyperglycemia-induced Glycosylation: A Driving Force for Vascular Dysfunction in Diabetes?
Authors: Hoffmann Brian R , Widlansky Michael E , Greene Andrew S
Published: June 2021

Vascular endothelial dysfunction leading to increased risk of coronary artery disease and peripheral vascular disease remains an important problem in type 2 diabetic mellitus (T2DM). T2DM patients with a high glycation index of hemoglobin A
1C
have a greater risk of vascular complications, suggesting there is a correlation between hyperglycemia-induced glycosylation and the progression of vascular dysfunction. The objective of this study was to better understand hyperglycemia-induced glycosylation in the vascular endothelium and its contribution to the onset and progression of T2DM. Initial characterization of the effects of hyperglycemia on
ex vivo
vasodilation and
in vitro
tube formation assays exhibited impaired endothelial phenotypes (p<0.05). PNGaseF enzymatic removal of endothelial cell surface glycosylations was able to restore the healthy tube formation phenotypes (p<0.05). To identify specific glycosylations contributing to this impairment we used a glycoproteomic approach to examine differential glycosylation on the rat microvascular endothelial cell surface following normal (4.5 mM) and high (25 mM) glucose treatments. In comparisons of the groups, 273 N-glycosylation and 499 O-glycosylation modified proteins were identified. Of these proteins, 87 were significantly increased in N-glycosylation and 143 in O-glycosylation as a result of the high glucose treatment (p<0.05). Bioinformatics analyses of the targets revealed differential glycosylation of important endothelial cell surface receptors, a diverse array of ion channels, and immune regulators. The CD59 glycoprotein, important for protection from innate immunity, was increased in N-glycosylation (2.02 fold, p<0.05) and O-glycosylation (unique). Interestingly, glycosylation of CD59 is known to inhibit this protection from innate immunity. The lipoprotein receptor LRP1, important for leptin signaling and energy homeostasis, was also significantly increased in total N-glycosylation (1.8-fold total; p<0.05) and has been linked to insulin resistance. Overall, this study identified multiple hyperglycemia-induced differential glycosylations in the endothelium that may be key regulators of vascular dysfunction during the onset and progression of T2DM.
Blood-Brain Barrier Crossing Renin-Angiotensin Drugs and Cognition in the Elderly: A Meta-Analysis
Authors: Ho Jean K. , Moriarty Frank , Manly Jennifer J. , Larson Eric B. , Evans Denis A. , Rajan Kumar B. , Hudak Elizabeth M. , Hassan Lamiaa , Liu Enwu , Sato Nobuyuki , Hasebe Naoyuki , Laurin Danielle , Carmichael Pierre-Hugues , Nation Daniel A.
Published: June 2021

Hypertension is an established risk factor for cognitive decline and dementia in older adults, highlighting the potential importance of antihypertensive treatments in prevention efforts. Work surrounding antihypertensive treatments has suggested possible salutary effects on cognition and neuropathology. Several studies have specifically highlighted renin-angiotensin system drugs, including AT1-receptor blockers and angiotensin-converting-enzyme inhibitors, as potentially benefiting cognition in later life. A small number of studies have further suggested renin-angiotensin system drugs that cross the blood-brain barrier may be linked to lower dementia risk compared to their nonpenetrant counterparts. The present meta-analysis sought to evaluate the potential cognitive benefits of blood-brain barrier crossing renin-angiotensin system drugs relative to their nonpenetrant counterparts. We harmonized longitudinal participant data from 14 cohorts from 6 countries (Australia, Canada, Germany, Ireland, Japan, United States), for a total of 12 849 individuals at baseline, and assessed for blood-brain barrier crossing potential within antihypertensive medications used by cognitively normal participants. We analyzed 7 cognitive domains (attention, executive function, language, verbal memory learning, recall, mental status, and processing speed) using ANCOVA (adjusted for age, sex, and education) and meta-analyses. Older adults taking blood-brain barrier-crossing renin-angiotensin drugs exhibited better memory recall over up to 3 years of follow-up, relative to those taking nonpenetrant medications, despite their relatively higher vascular risk burden. Conversely, those taking nonblood-brain barrier-penetrant medications showed better attention over the same follow-up period, although their lower vascular risk burden may partially explain this result. Findings suggest links between blood-brain barrier crossing renin-angiotensin drugs and less memory decline.
Urinary Heavy Metals and Longitudinal Changes in Blood Pressure in Midlife Women: The Study of Women’s Health Across the Nation
Authors: Wang Xin , Karvonen-Gutierrez Carrie A. , Herman William H. , Mukherjee Bhramar , Harlow Sioban D. , Park Sung Kyun
Published: June 2021

Environmental exposure to heavy metals may contribute to increased blood pressure; however, evidence from midlife women who are at greater risk of cardio-metabolic disease is limited. We evaluated the associations of urinary concentrations of arsenic, cadmium, mercury, and lead with longitudinal changes in blood pressure in the Study of Women’s Health Across the Nation Multi-Pollutant Study. The sample included 1317 White, Black, Chinese, and Japanese women, aged 45 to 56 years at baseline (1999–2000), whose systolic blood pressure (SBP) and diastolic blood pressure were measured annually or biannually through 2017. Urinary metal concentrations were determined at baseline. Longitudinal changes in SBP and diastolic blood pressure were modeled using linear mixed-effects models by tertiles of metal concentrations. After multivariable adjustment, estimated annualized increases (95% CI) in SBP in the highest and lowest tertiles were 0.93 (0.85–1.01) mm Hg and 0.74 (0.66–0.82) mm Hg for arsenic, 0.82 (0.75–0.90) mm Hg and 0.72 (0.65–0.80) mm Hg for mercury, and 0.86 (0.78–0.93) mm Hg and 0.72 (0.64–0.79) mm Hg for lead, respectively. Similar results were observed for associations of arsenic, mercury, lead with diastolic blood pressure. Urinary cadmium was associated with a greater rate of increase in SBP only among never smokers. Women with higher concentrations of all four metals had greater annualized increases in SBP and diastolic blood pressure than those with lower concentrations. Our findings suggest that exposure to heavy metals may accelerate the increase in blood pressure in midlife women, supporting the need for continued efforts to reduce these environmental exposures.
Abstract 397: Regional Differences in Periaortic Adipose Tissue
Authors: Tran Khanh-Van , Fitzgibbons Timothy , Corvera Silvia
Published: June 2021

Introduction:
To combat obesity and associated comorbidities, it is vital to have an in-depth understanding of the adipose tissue. The presence of brown adipose tissue is associated with decrease risk of T2DM while the opposite is true for the presence of visceral white adipose tissue. In this study, we examined the adipocyte populations surrounding the thoracic and abdominal aorta of C57Bl/6J mice.

Methods:
We utilized the aortic ring assay to examine the adipocytes arising from the thoracic and abdominal regions of the aorta. We previously found that adipocyte progenitors proliferate when tissue is embedded in Matrigel. C57Bl/6J mice were sacrificed, thoracic and abdominal aorta fragments were removed, embedded in Matrigel matrix and allowed to proliferate in the absence or in the presence of Rosiglitazone, a PPARγ agonist. Explants were then fixed in formaldehyde for immunofluorescence analysis or used for RNA extraction and subsequently real-time PCR.

Results:
Classically, cells that arise from the aorta fragments are spindle-like and express several endothelial cell markers, such as CD31, CD34 and vWF. We found that in the presence of rosiglitazone, cells arising from the aorta accumulate lipid droplets to take on an adipocyte phenotype. In the presence of rosiglitazone, the expression of CD31 and CD34 are decreased and expressions of adiponectin and perilipin are increased. The perilipin staining patterns in these cells are consistent with that of adipocytes, i.e. surrounding a lipid droplet. Adipocytes around the abdominal aorta have higher mRNA levels of Hoxc9, a homeobox gene expressed higher in white adipocytes than brown adipocytes. Conversely, adipocytes arising from the thoracic aorta have higher expression of UCP-1 and cidea, markers of brown adipocytes.

Conclusion:
Results show that adipocytes from thoracic and abdominal regions of the aorta have different genotypes and phenotypes. Cells surrounding the thoracic aorta may have more of a “beige,” in between white and brown phenotype, as opposed to a classic white adipocyte phenotype seen in fat cells surrounding the abdominal aorta. Our e
x vivo
aortic ring model is a tool that can provide valuable insight into the role that adipocytes play in different regions of the aorta.
Abstract 128: TICI Reperfusion in HERMES: Success in Endovascular Stroke Therapy
Authors: Liebeskind David S , Jovin Tudor G , Majoie Charles B , Mitchell Peter J , van der Lugt Aad , Menon Bijoy K , San Román Luis , Campbell Bruce C , Muir Keith W , Hill Michael D , Dippel Diederik W , Saver Jeffrey L , Demchuk Andrew , Dávalos Antoni , White Philip , Brown Scott , Goyal Mayank ,   for the HERMES Collaborators
Published: June 2021

Background:
Successful revascularization after endovascular therapy for acute ischemic stroke is measured by TICI score, yet variability exists in scale definitions and use. We examined the degree of reperfusion and association with outcomes in the HERMES collaboration of recent endovascular trials.

Methods:
An independent reader of the HERMES Imaging Core, blind to all other data, evaluated the angiography of subjects treated with endovascular therapy in HERMES. A battery of various TICI scores (mTICI, oTICI, oTICI2C) was used to define reperfusion of the initial target occlusion on noninvasive imaging (ITO) and conventional angiography (CATO). Statistical analyses examined all TICI reperfusion metrics and correlation with clinical outcomes.

Results:
Angiography of 593 subjects was analyzed, including ITO (124 ICA, 413 M1, 47 M2) and CATO (161 ICA, 329 M1, 62 M2). Across the entire scale range (0-3), the mTICI (AUC 0.61), oTICI (AUC 0.61) and oTICI2C (AUC 0.62) revealed similar ROC characteristics (p=0.450) in discriminating that more reperfusion is associated with better clinical outcomes. Using oTICI2C (3=100%, 2C=90-99%, o2B=67-89%, m2B=50-66%) of CATO, there were 44 TICI 3 (8%), 125 TICI 2C (22%), 178 TICI o2B (32%), 80 TICI m2B (14%), 85 TICI 2A (15%), 15 TICI 1 (3%) and 35 TICI 0 (6%). mRS shift analyses from baseline to 90 days revealed increasing TICI grades were linked with better outcomes (Figure), with significant distinctions of m2B vs. 2C (p=0.023) and all 2B combined vs. 3 (p=0.045).

Conclusions:
The benefit of endovascular therapy in HERMES was strongly associated with increasing degrees of TICI reperfusion. The oTICI2C metric reveals important distinctions in clinical outcomes that should be used in future studies.
Abstract 207: A Systematic Literature Review of Patients with Carotid Web and Acute Ischemic Stroke
Authors: Dhruv Parth , Kim Jae , Koffel Jonathan , Anderson David , Streib Christopher
Published: June 2021

Introduction:
Carotid Web (CW) is a rare form of focal fibromuscular dysplasia that results in an abnormal shelf-like projection of intimal fibrous tissue into the carotid bulb. It is theorized that CW leads to ischemic stroke secondary to blood flow stasis and subsequent embolization. There is uncertainty in diagnosis, prognosis, and optimal management of this uncommon entity. To address this knowledge gap, we performed a systematic literature review (SLR) of CW.

Methods:
Our literature search for CW and related terms yielded 1017 results. After a preliminary assessment of all 1017 retrieved manuscripts; 72 manuscripts were reviewed in detail. A total of 31 manuscripts met entry criteria and were included in our SLR. We present the demographics, cardiovascular (CV) risk factors, neuroimaging findings, stroke recurrence or stroke free-duration, and treatment modality of CW patients.

Results:
Our SLR resulted in 104 patients with CW, (median age at presentation: 45.7 [IQR 45-57], 52.3% female). The majority (68.3%) of CW patients did not have CV risk factors, however, 22.2% of patients were smokers. 47.8% of patients suffered recurrent stroke. The majority of patients were ultimately treated with antiplatelet therapy (94.4%) and 21.2% underwent carotid revascularization (4.8% carotid stenting, 16.3% carotid endarterectomy). None of the patients who underwent revascularization had a reported stroke recurrence. See Table.

Conclusions:
CW is a rare disease leading to ischemic stroke in younger patients without typical CV risk factors. CW patients in our SLR were at extremely high risk for recurrent stroke (47.8%). Aggressive secondary stroke prevention measures are indicated in this patient population, however, the optimal treatment strategies remain unclear. Carotid revascularization may be the definitive treatment for certain patients with CW, but further studies are needed as incomplete reporting and potential publication bias limit our findings.
Abstract WP272: Impact of Stroke Legislation on Developing Stroke Systems of Care and Improving Acute Therapy
Authors: Kim Elizabeth , Jones Peggy , Richards Christopher T , Prabhakaran Shyam , Love Tracy , Cranick Lesley , O’Neill Kathleen , Sednew Renee , Scharnott Michelle , Peysakhovich Mark , Meixner Alex , Mirostaw Julie , Alberts Mark J
Published: June 2021

Background:
Stroke is the 4th leading cause of death in Illinois and the leading cause of disability. In 2009, Illinois passed stroke legislation to establish a Stroke Advisory Sub-committee to advise the State EMS Advisory Council. Legislation also created 11 multidisciplinary EMS Regional Stroke Advisory Subcommittees, recognition of certified Primary Stroke Centers, state designation of Emergent Stroke Ready hospitals and updating of EMS routing protocols. In 2014, updated legislation was passed to include state recognition of nationally certified Comprehensive Stroke Centers (CSC), updating of EMS routing protocols to include CSC’s and establish a state stroke fund created from hospital designation fees.

Hypothesis:
Implementation of Illinois’s stroke legislation by EMS region enhances systems of care collaboration between hospitals and EMS providers and improves timely intervention for patients with acute ischemic stroke.

Methods:
We queried the state-wide AHA’s Get With The Guideline stroke registry, which captured 72% of Illinois stroke discharges from 2009 through 2015. In total, 90 hospitals entered data, 57 from PSCs, 8 from CSCs, and 25 from Emergent Stroke Ready hospitals. All patients included in the registry as confirmed stroke were included for analysis. Hospital time and treatment data were recorded in an aggregate manner. Descriptive statistics were used to report results.

Results:
The total number of confirmed stroke patients treated was 123,859 Median door-to-needle (DTN) times for IV tPA therapy went from 85 minutes in 2009 to 56 in 2015 . The percent of acute ischemic stroke patients with a door-to-needle time of 60 minutes or less increased from 18.0% in 2009 to 62.9% in 2015.

Conclusions:
The decrease in DTN from 2009 to 2015 is an indicator of improved care due to the Illinois stroke system of care model. This is a good example of all system stakeholders including state government and EMS working collaboratively to improve care. Which elements of a stroke system are responsible for this improvement (hospital designation, routing, EMS training) will require further research. Regional coordination of resources and state hospital designation may impact prehospital routing of suspected strokes and may improve patient outcomes.
Abstract 80: Predictors of Early Infarct Recurrence in Patients With Symptomatic Intracranial Atherosclerotic Disease
Authors: Prabhakaran Shyam , Liebeskind David S , Cotsonis George , Nizam Azhar , Feldmann Edward , Sangha Rajbeer , Campo-Bustillo Iszet , Romano Jose
Published: June 2021

Introduction:
Prior studies have evaluated risks factors for recurrent stroke in patients with symptomatic intracranial atherosclerotic disease (ICAD). However, few reports have assessed risk factors for early infarct recurrence in the territory distal to the symptomatic artery.

Methods:
We analyzed data from patients who underwent study-paid brain MRI at 6-8 weeks after enrollment in the ongoing MyRIAD study, an NIH/NINDS funded prospective multicenter observational study of patients with recent (
<
21 days) stroke or TIA (recurrent or with DWI) caused by ICAD 50-99% without planned angioplasty/stenting. The outcome of interest was new infarcts on brain MRI (on DWI or FLAIR) at 6-8 weeks compared to qualifying brain MRI at time of index stroke or TIA. Qualifying events and clinical and radiographic outcomes are centrally ascertained by 2 independent reviewers. We used logistic regression to identify independent clinical predictors of new infarct in the territory of the symptomatic artery.

Results:
Among 84 (80%) of 105 enrolled patients in MyRIAD with 6-8 week MRI, the mean age was 63.6
+
12.4 years, 83.1% have stenosis 70-99%, and 51.2% had history of diabetes; those who underwent MRI did not differ from those who did not undergo MRI. A new DWI/FLAIR infarct in the territory of the symptomatic artery was noted in 26.2%. Those with recurrent infarcts were younger (57.7 vs. 65.7 years, p=0.009), more likely to have diabetes (71.4% vs. 44.3%, p=0.043), have greater degree of stenosis (82.5% vs. 76.0%, p=0.099), and have greater decline in systolic blood pressure (SBP) from enrollment to 6-8 week follow-up (+5 vs. -6.2 mm Hg, p=0.074). In adjusted analyses, age (aOR 0.922, 95% CI 0.869-0.979) and change in SBP (aOR 0.967, 95% CI 0.937-0.997) were related to new infarct in the territory.

Conclusions:
Early recurrent infarcts occur in more than one-quarter of patients with symptomatic ICAD, may be a suitable biomarker of disease activity, and add to the subclinical burden of this high-risk disease. Given the association between SBP lowering and infarct recurrence, studies of early blood pressure management strategies, including extended permissive hypertension, may be warranted in this population.
Abstract WP315: Neuroprotective Effects of Endogenous Adropin in Experimental Ischemic Stroke
Authors: Yang Changjun , Sanz Brian D , DeMars Kelly M , Butler Andrew A , Candelario-Jalil Eduardo
Published: June 2021

Adropin is an endogenous peptide highly expressed in the brain and is encoded by the energy homeostasis-associated gene,
Enho
. We recently found that treatment with synthetic adropin peptide reduces infarct volume in ischemic stroke. This protection by exogenous adropin is associated with a significant increase in endothelial nitric oxide synthase (eNOS) phosphorylation (Ser
1176
) and reduction of blood-brain barrier (BBB) permeability. However, it is not known whether endogenous adropin modulates ischemic brain damage. We hypothesize that lack of brain adropin increases stroke damage by exacerbating neurovascular dysfunction, while overexpression of this peptide decreases ischemic brain injury. We measured infarct size and BBB damage in male and female adropin overexpressing transgenics (AdrTg), adropin knockout (
Enho
-/-
), and corresponding wild-type (WT) control mice subjected to permanent middle cerebral artery occlusion (pMCAO). At 48h after stroke, infarct volume was significantly smaller in AdrTg mice of both sexes compared to WT controls, while stroke resulted in a much larger infarction in both male and female
Enho
-/-
compared to
Enho
+/+
mice. Brain levels of IgG and albumin, two sensitive markers of BBB disruption, were significantly reduced in AdrTg mice compared to the WT group, while the levels of these two plasma proteins in the ischemic brain of
Enho
-/-
mice were dramatically increased compared to
Enho
+/+
animals after stroke. Latex vessel casting showed no differences in cerebrovascular anatomy or the diameter of the main brain arteries between
Enho
-/-
, AdrTg and their respective WT controls. In AdrTg mice, we found a positive correlation between brain adropin levels and eNOS phosphorylation. These data suggest that increased eNOS phosphorylation by adropin is a potential mechanism underlying its protective effects in stroke. Consistent with this hypothesis, the protective effects of adropin were completely abolished in eNOS knockout (eNOS
-/-
) mice subjected to pMCAO. Taken together, our findings show for the first time that endogenous adropin is a neuroprotective peptide in ischemic stroke. Understanding adropin signaling and function could offer a novel therapeutic strategy for the treatment of ischemic brain damage.
From Competition to Cooperation: The Radicalization Effect of Salafists on Egypt’s Muslim Brotherhood in the Aftermath of the 2011 Uprising
Authors: Yaghi Mohammad
Published: June 2021

Using the case studies of the 2012 Constitution, the call of al-Jabha al-Salafiyya for the Revolution of the Muslim’s Youth (rmy) and the Salafi’s statement of Nida Ard al-Kinana, this article provides empirical evidence that the Salafists have a radicalization effect on Egypt’s Muslim Brotherhood when they compete or cooperate with each other. By “radicalization effect,” the article means pushing the Brotherhood to build less inclusive institutions and/or pulling them toward the justification of the use of violence in religious terms in their confrontation with al-Sisi’s regime. Methodologically, the article relies on the Salafist and the Brotherhood statements as well as on the work of other scholars.
Contribution to Chaetosphaeriaceae—Kionochaeta setosimplicia sp. nov. from Thailand
Authors: NUANKAEW SALILAPORN , CHUASEEHARONNACHAI CHARUWAN , SOMRITHIPOL SAYANH , KWANTONG PAPICHAYA , SRIHOM CHARISA , CHANDAPHEAKDEY ROS , BOONYUEN NATTAWUT
Published: June 2021

Species of Kionochaeta are saprobes commonly associated with decaying plant materials. As a part of surveys on freshwater microfungi, Kionochaeta-like taxon was found on a submerged decaying twig at a rivulet in the protected forest area of Khao Kheow Open Zoo (KKOZ), eastern Thailand. This taxon is microscopically similar to Kionochaeta, but clearly differs by its accessory lateral setiform branches and sterile setae arising beside the conidiophores. Multi-locus phylogenies inferred from four combined loci (SSU, ITS, LSU and RPB2) demonstrated that this taxon clustered as a unique clade with Kionochaeta species (Chaetosphaeriaceae, Chaetosphaeriales). Therefore, we introduce this taxon as K. setosimplicia, and it is phylogenetically close to K. castaneae, K. ramifera and K. pughii. Kionochaeta setosimplicia is separated from K. microspora and K. spissa. In Kionochaeta clade 1, K. nanophora forms a basal taxon. Kionochaeta ivoriensis forms a distinct Kionochaeta clade 2 distant from other Kionochaeta.
Assessment of Bone Regeneration Using Tooth Ash and Injectable Platelet Rich Fibrin: A Microcomputed Tomographic (CT) Analysis
Authors: Alshihri Saleh , Kindi Mohammed , Alfotawi Randa , Hindi Marium Al , Alghamdi Osama , Ramalingam Sundar , Assiri Mohammed
Published: June 2021

Introduction: One of the main challenge of bone graft or socket preservation in particular is to get good quality and quantity of bone in short time prior to implant bed preparation. The buccal bone at the crest of the ridge is a very thin bone and usually resorb faster than
the rest of alveolar bone which may hinder an optimum dento-osseous implant insertion. The purpose of this study will be to assess the bone regeneration capabilities of Tooth Ash Particles (TAP) with injectable Plasma Rich Fibrin (i-PRF) and Tooth Ash Particles (TAP) alone at defects created
in the goat mandible bone using micro-computed tomographic (micro-CT). Materials and Methods: A total, 54 bone defect (5 mm × 8 mm) were performed in the 18 goats. The created defect received different treatment (Tx): Tx.A: Unfilled defect (allow natural bone regeneration; Tx.B:
Tooth Ash particle (TAP) alone; Tx.C: Tooth Ash + injectable PRF (TAP/i-PRF). Six goats, were sacrificed at different time points:Group 1: at 2nd week, Group2: at 5th week and Group3 at 8th week. The newly formed bone (NFB) was analyzed using micro-CT at different time points. Quantitative
and qualitative assessment were carried out namely; the volume of new bone formation (NF-BV) within the defect and its mineral density (NF-BMD), Trabecular Thickness (Tb Th), Trabecular Number (Tb N) and Trabecular Separation (Tb Sp). Result: By 8th week, the mean NF-BV was 69.482 ±
6.554 mm3 (cubic millimeter), 65.872±6.804 mm3, 26.820±14.643 mm3, while the mean NF-BMD was 0.417±0.119g/mm3, 0.786±0.036 g/mm3, 0.805±0.033 g/mm3 for the defects which received Tx.C, Tx.B
and Tx.A respectively. At 8th weekTb Th of NFB was 0.612±0.168, 0.913±0.112, and 0.701 ±0.126, Trabecular Number of NFB was 2.062±0.946, 1.002±0.155, and 1.816±2.042 and, Trabecular Separation of NFB was 0.330 ±0.131, 0.559 ±0.110, and
0.495 ±0.258 for the defects which received Tx.A, Tx.B and Tx.C respectively. Conclusion: Micro-CT study demonstrated that tooth ash particles mixed with injectable Platelet Rich Fibrin (i-PRF) on mandibular bone defect in goat’s model, resulting in new bone with significantly
higher volume, mineral density and less remodeling rate when compared with normal bone regeneration of unfilled defects.
A Shunt Method for the Treatment of Syringomyelia Secondary to Trauma and Tuberculous Meningitis
Authors: Li Hao , Lou Yongli , Liu Yong
Published: June 2021

The pathogenesis and surgical treatment of syringomyelia (SM) secondary to trauma and tuberculous meningitis are not clear. We hypothesize that the main cause of SM is impaired cerebrospinal fluid circulation in the subarachnoid space due to spinal arachnoid adhesion induced by trauma
or tuberculous meningitis. Using trans-segmental subarachnoid shuntsurgery, we re-established normal cerebrospinal fluid circulation in the subarachnoid space, which resolved the potential cause of SM formation. This study aimed to evaluate the efficacy of trans-segmental subarachnoid shunt
surgery in the treatment of SM secondary to trauma or tuberculous meningitis. A total of 143 patients with SM after trauma and tuberculous meningitis were recruited between September 2014 and February 2017. Among these patients, 64 had posttraumatic SM, including 39 males and 25 females with
an age range of 21–65 years and an average age of 40.02 years, and 79 patients had SM secondary to tuberculous meningitis, including 49 males and 30 females with an age range of 23–62 years and an average age of 44.95 years. All patients underwent trans-segmental subarachnoid shunt
surgery and were subjected to magnetic resonance imaging (MRI) examination between 6 months and 1 year and between 2 and 2.5 years after surgery. Clinical efficacy was assessed by the Tator method. The main symptoms or signs were evaluated by improvement, stabilizationor aggravation of symptoms.
Patients were followed-up twice, once at 6 months to 1 year after surgery and once at 2 to 2.5 years after surgery. A total of 143 patients were followed-up for the first time, among whom, the clinical symptoms were improved in 59 patients(41.26%), stable in 51 patients (35.66%) and aggravated
in 36 patients (23.08%). MRI exaination showed that the spinal cord cavities completely disappeared in 27 patients (18.88%), shrank significantly in 71 patients (49.65%), did not change or shrink significantly in 32 patients (22.38%) and expanded in 13 patients (9.09%). A total of 122 patients
were followed-up for the second time, and the clinical symptoms were improved in 69 patients (56.56%), stable in 25 patients (20.49%) and aggravated in 28 patients (22.95%). Compared to the condition in the first follow-up, 14 patients who were stable were improved, 5 patients who were aggravated
were improved, and the symptoms did not change significantly among other patients. The imaging examination did not show any significant change compared to that of the first follow-up. We suggest that trauma or tuberculous meningitis can induce spinal arachnoid adhesion, leading to impaired
cerebrospinal fluid circulation in the subarachnoid space, which then causes SM. Trans-segmental subarachnoid shunt surgery is a noninvasive, safe and effective treatment for SM secondary to trauma and tuberculous meningitis.
Galangin Alleviates Tumor Progression and Metastasis in Intraperitoneal Ovarian Cancer Model via Inhibiting Janus Kinase 1/Signal Transduction and Activator of Transcription 3 Signaling
Authors: Yue Hua , Zhang Xiuling , Xia Yali
Published: June 2021

To investigate the impact of galangin on tumor progression and metastasis in intraperitoneal ovarian cancer model. Ovarian cancer cells were treated with DMSO or galectin, cell viability was detected by MTS or acid phosphatase assay, SKOV3 cells were transfected with STAT3 targeted shRNA and the expression of signal transduction-related proteins in cells was analyzed by immunoblotting assay, the expression of IL-6, IL-2, INF-y was estimated by enzyme-linked immunosorbent assay the peritoneal metastasis model of ovarian cancer was established using shSTAT3 transfected or untransfected SKOV3 cells and treated with galangin or DMSO. Tumor mass, number of small tumor nodules and ascites volume were detected in the mouse model. Ovarian cancer-bearing mice treated with galangin showed a dramatic decreased tumor burden as demonstrated by the 25 times-reduced total weight of small tumor nodules, 60%-reduced primary tumors, attenuated luciferase activity and completely blocked ascites production. Moreover, galangin inhibited cell viabilityin vitroin a concentration-dependent manner. Further, p-STAT3 was suppressed by galangin treatment bothin vivoand vitro. Galangin inhibited the expression of p-JAK1, the upstream signaling of p-STAT3 and IL-6 in the downstream. Meanwhile, knockdown of STAT3 by shSTAT3 transfection mimicked the therapeutic effects of galanginin vivoand vitro. Galangin supresses IL-6 secretion, peritoneal metastasis and ascites production by inhibiting JAK1/STAT3 signaling.
Toxicities of high-dose chemotherapy and autologous hematopoietic cell transplantation in older patients with lymphoma
Authors: Dahi Parastoo B. , Lee Jasme , Devlin Sean M. , Ruiz Josel , Maloy Molly , Rondon-Clavo Carlos , Petrlik Erica , Tamari Roni , Shah Gunjan , Scordo Michael , Matasar Matthew J. , Hamlin Paul A. , Papadopoulos Esperanza , Jakubowski Ann A. , Perales Miguel-Angel , Moskowitz Craig H. , Sauter Craig S. , Giralt Sergio A.
Published: June 2021

High-dose chemotherapy and autologous hematopoietic cell transplantation is an effective consolidation therapy in lymphoma; however, its use in elderly patients has been limited because of concerns for greater toxicity in this group. We investigated the toxicities of carmustine, etoposide, cytarabine, and melphalan (BEAM) and autologous hematopoietic cell transplantation (AHCT) in 346 patients in 2 age groups: 279 patients aged 60 to 69 years and 67 patients aged ≥70 years. The majority developed severe toxicities; the most common were febrile neutropenia, gastrointestinal, infections, and cardiovascular. Older patients were at higher risk for grade ≥3 cardiovascular toxicities (hazard ratio [HR], 3.36; 95% confidence interval [CI], 2.25-5.00; P < .001) and skin toxicities (HR, 2.45; 95% CI, 1.08-5.54, P = .032). In the older group, nonrelapse mortality at 100 days and at 2 years was 2.99% (95% CI, 0.55-9.32) and 6.2% (95% CI, 1.97-13.95), respectively, vs 1.79% (95% CI, 0.68-3.92) and 2.91% (95% CI, 1.37-5.42), respectively, in the younger group. When adjusting for the number of grade ≥3 toxicities within the first 100 days, older patients had a 1.71-fold (95% CI, 1.08-2.71) increased risk for progression or death relative to younger patients. Although BEAM followed by AHCT is effective, it is associated with significant organ toxicities, especially in patients aged ≥70 years. Interventions to mitigate toxicities while maintaining efficacy are much needed.
Revised manuscript R2, clean version are serum levels of 25-hydroxy vitamin D reduced following orchiectomy in testicular cancer patients?
Authors: Dieckmann Klaus-Peter , Andura Osama , Pichlmeier Uwe , Otte Klaus Martin , Isbarn Hendrik , Wülfing Christian
Published: June 2021

Background

The testis represents one place where the progenitor of vitamin D is converted into its active form. Loss of one testis was suggested to result in reduced vitamin D serum levels. Vitamin D deficiency would represent a significant health problem in the long-term course of patients with testicular germ cell tumors (GCTs) since most of them survive. The purpose of this study was to look to the serum 25(OH)-Vitamin D (25OHD) levels in patients with GCTs before and after orchiectomy. A total of 177 GCT patients underwent measurements of serum 25OHD levels, thereof 83 with preoperative measurements and 94 with measurements at six particular time-points from immediate postoperatively to >24 months. Longitudinal assessments of 25OHD serum levels were performed in individual patients with repeated measurements. A second analysis involved patient cohorts with measurements at six postoperative time-points. Serum levels of patients were also compared with 2 control groups, one consisting of 84 patients with non-neoplastic testicular diseases and another with 237 patients with non-neoplastic urologic diseases. We also looked to associations of 25OHD levels with levels of testosterone, follicle stimulating hormone (FSH), age, histology of GCT and season. Descriptive statistical methods were employed to compare groups and to analyze changes over time.

Results

Normal serum levels of 25OHD were found in 21.7%, 23.1%, 20.2%, 21.9% in GCT patients preoperatively, after >2 years, in control group 1 and control group 2, respectively. Levels were significantly higher in spring and summer, but no association was found with other parameters. We found a significant transient decrease of 25OHD levels with a nadir at 6-12 months after orchiectomy and a recovery thereafter.

Conclusion

Contrasting with previous studies we found no permanent reduction of serum 25OHD levels after orchiectomy but transient postoperative drop of 25OHD levels. There were no associations of 25OHD levels with age, and levels of testosterone or FSH. Our results may point to a particular role of the testis in vitamin D metabolism and may thus enhance the understanding of the diverse physiological roles of the testis.