bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P200: Role of Macula Densa Neuronal Nitric Oxide Synthase in Control of Renin Release and Blood Pressure Recovery Following Hemorrhagic Shock
Authors: Wang Lei , Wang Shaohui , Buggs Jacentha , Zhang Jie , Wei Jin , Zhang Gensheng , Liu Ruisheng 
Published: June 2021

Renin is a rate limiting factor for generation of angiotensin II, which is essential for blood pressure regulation. The role of macula densa nitric oxide (NO) in renin release is not conclusive. The goal of this study was to elucidate the role of macula densa neuronal NO synthase (NOS1) in control of renin release in response to sodium challenges and hemorrhagic shock, as well as in blood pressure recovery after hemorrhagic shock.
C57BL/6L mice and macula densa specific NOS1 knockout (MD-NOS1KO) mice were given 10 days of low (0.1% NaCl), normal (0.4% NaCl) and high (1.4% NaCl) sodium diet. Hemorrhagic shock was induced by withdrawing 0.4 ml whole blood from the right retro-orbital sinus. Mean arterial pressure (MAP) in conscious mice was monitored by radio-telemetry system. Plasma renin concentration (PRC) was determined by radioimmunoassay.
Low sodium diet stimulated PCR by 29% (from 685 ± 32 to 883 ± 112 ng/ml/hr) in WT mice and by 16% (from 652 ± 24 to 756 ± 124 ng/ml/hr) in the MD-NOS1KO mice (n=5/group, p<0.01 vs WT). PCR was not significantly different between the WT and MD-NOS1KO mice fed a normal or high salt diet. As shown in Fig1A, following removal of 0.4ml of blood, MAP dropped to about 40mmHg in the WT mice and 35mmHg in the MD-NOS1KO mice. MAP recovered faster in WT mice than the MD-NOS1KO mice. In Fig1B, PRC increased over 200% of the basal value in WT mice, but only increased about 26% in the MD-NOS1KO mice (n=4/group, p<0.01 vs WT).
We conclude that NOS1 in the macula densa facilitates renin release. Lack of macula densa NO generation impairs blood pressure recovery, which may be mediated by limiting renin release during hemorrhagic shock.
Abstract P282: Use of a CRISPR/Cas9 System for Specification of the Renin Cell Phenotype
Authors: Medrano Silvia , Brown Robin Isadora , Martinez Maria F , Sequeira-Lopez Maria Luisa S , Gomez R A 
Published: June 2021

Renin is a key enzyme/hormone that controls blood pressure and fluid-electrolyte homeostasis. Renin transcription is subject to complex developmental, physiological, and pathological regulation. We have identified the cAMP pathway and associated epigenetic marks as crucial regulators of renin expression. However, those studies involved the use of cAMP activators or analogues or histone deacetylase inhibitors that alter the transcriptome and epigenome of the whole cell without exclusively targeting the renin locus.
The CRISPR/Cas9 Type II bacterial immune system has been modified for genome editing in eukaryotes. This system consists of a CRISPR-associated endonuclease (Cas9) and single guide RNA (sgRNA) to target specific genomic areas. Modifications to the Cas9 enzyme have allowed the use of CRISPR to regulate gene expression by targeted modification of epigenetic marks.
In this study we tested whether renin expression can be induced by a nuclease-null dCas9 protein fused to the catalytic core of the acetyltransferase p300 (dCas9p300) and sgRNAs directed to the renin promoter and enhancer.
We used cultured arteriolar smooth muscle cells of the renin lineage that constitutively express CFP (a renin lineage marker) and YFP only when the renin gene is turned on. Cells were transfected with a dCas9p300 and sgRNA expression vectors and analyzed for YFP expression 48h after transfection.
We tested four sgRNAs targeting the renin enhancer and five sgRNAs targeting the renin promoter either individually or in combination. We found few YFP+ cells when all 4 enhancer sgRNAs or 5 promoter sgRNAs were simultaneously used. The highest YFP expression (32±9 cells/well) was observed when two enhancer sgRNAs (at positions -2,757 and -2,631) and three promoter sgRNAs (at positions -719, -50 and -25) were simultaneously added to the cells. We did not find any YFP+ cells when the dCas9p300 plasmid was transfected without sgRNAs or when cells were transfected with a control plasmid.
Our data support targeted acetylation as a causal mechanism of renin transactivation. CRISPR/Cas9 provides a tool to study the regulation of renin expression by targeting epigenetic marks in the promoter and enhancer of renin.
Abstract P283: Effects of Ovarian Estrogens on Local RAS Activity in Cardiomyocytes and Non-cardiomyocytes
Authors: Sun Xuming , Ahmad Sarfaraz , Lin Marina , Zapata-Sudo Gisele , Cheng Cheping , Varagic Jasmina , Ferrario Carlos , Wang Hao , Groban Leanne 
Published: June 2021

The relatively low efficacy of ACE-Is in the treatment of heart failure (HF) in women after estrogen (E2) loss may be due to their inability to reach the intracellular sites at which Ang II is generated and/or the existence of cell-specific mechanisms in which ACE is not the essential processing pathway for Ang II formation. We compared the metabolic pathway for Ang II formation in freshly isolated cardiomyocytes (CMs) and non-cardiomyocytes (NCMs) extracted from hearts of gonadal-intact and ovariectomized (OVX) adult WKY and SHR rats. Circulating levels of angiotensinogen (AOGEN) were higher in WKY (958 ± 47 pg/mL) vs. SHR (SHR: 626 ± 40 pg/mL; P<0.05 strain effect) and E2 loss augmented this effect in WKY (WKY OVX: 1,169 ± 66 pg/mL vs. SHR OVX: 625 ± 41 pg/mL). Correspondingly, plasma Ang II levels were higher in WKY vs. SHR (strain effect: WKY: 62 ± 6 pg/mL vs. SHR: 42 ± 9 pg/mL), independent of OVX. Chymase activity was nearly 40-fold higher in NCMs compared to CMs, and for the NCMs, activities were highest in cells from WKY vs. SHR and OVX vs. intact rats (P<0.05 strain and E2 effects, respectively) (Figure).
Neither strain nor gonad status influenced the lower ACE activity found in both NCMs and CMs. In contrast, ACE2 activity in CMs and NCMs was higher in cells from WKY vs. SHR (P<0.05 strain effect), independent of E2 status. We conclude that NCMs from WKY and SHR express significantly higher levels of chymase, ACE, and ACE2 activities. E2 loss leads to selective changes in the activity of chymase, but not ACE, in NCMs. The significance of these novel findings is that targeted cell-specific chymase rather than ACE inhibition may have a greater benefit in the management of HF in women after menopause.
Abstract P308: Pilot Study: 24 Hour and Daytime Blood Pressure Lowering Effect of Melatonin in Younger and Older Non-hypertensive Adults
Authors: Cheng Cynthia , Gooneratne Nalaka S , Payton Colleen , Keith Scott , Hafycz John 
Published: June 2021

The study objective was to study the effect of melatonin on blood pressure (BP) in both younger and older normotensive and prehypertensive individuals.
Preliminary data available at the time of abstract submission are presented for 12 participants (10 ≤ 50 years old; two ≥ 60). free of known hypertension or diabetes. All subjects had office and 24 hour ambulatory BP measured before and after dosing of 9 mg controlled release melatonin for six weeks.

Subjects are described in
Table 1
. Full data analysis for all 30 pilot subjects, including 12 older and 10 control subjects, will be available for the presentation.

Table 2
shows results comparing baseline and follow-up BP. Mean 24 hour SBP/DBP, as well as ambulatory daytime SBP/DBP, were significantly lower following melatonin administration.

In conclusion, these findings suggest possible future utility of melatonin for lowering BP in both younger and older non-hypertensive individuals.
Abstract P336: Changes in Arterial Pulse Wave Velocity, Effective Arterial Elastance, and Contractility in Hypertension Induced Cardiovascular Disease in Rats: Independent Prognostic Values
Authors: Roof Steve R , del Rio Carlos 
Published: June 2021

Hypertension is an established comorbidity in cardiovascular (CV) disease. Changes in aortic pulse-wave velocity (PWV) have been shown to predict the longitudinal development of hypertension, its underlying vascular alterations, and associated CV risks. However, as the heart and arterial tree are coupled, it has been shown that the successful monitoring of not only CV disease progression, but of its therapies, requires the evaluation of ventriculo-arterial uncoupling and its determinants. This study aimed to establish the relationships between PWV and changes in the estimated effective arterial elastance (Ea) and/or contractility over a wide-range of hypertension-induced CV pathologies in a controlled rodent model/environment.
In a large series (n = 96) of anesthetized (pentobarbital) male Sprague-Dawley rats, PWV (carotid-to-femoral), systolic pressures, contractility (preload recruitable stroke work; PRSW), and Ea were evaluated using echocardiography, invasive hemodynamics, and left ventricular (LV) pressure-volume relationships. Data were collected in healthy animals (n = 37) and those with hypertension-induced chronic systolic/diastolic left-ventricular dysfunction (n = 59) (chronic beta-adrenergic stimulation and/or renoprival). A subset of each group, were also studied under common clinical cardio/vasoactive pharmacological interventions.

Disease animals had significantly (P < 0.001) higher systolic pressures (141 ± 4 vs 116 ± 4 mmHg), faster PWV (9.0 ± 0.8 vs 5.1 ± 0.5 m/s), and depressed contractility (PRSW: 39 ± 1 vs. 49 ± 1 mmHg*). Both PWV (7.6 ± 0.7 to 4.3 ± 0.4 m/s) and Ea (95 ± 10 to 63 ± 8 mmHg/mL) decreased in response to vascular therapies in the diseased animals; only PWV was reduced in healthy rats. More interestingly, over all conditions, PWV was a poor predictor of both systolic pressures (R
2
=0.00009) and Ea (R
2
= 0.024). but yet was a moderate predictor of PRSW (R
2
=0.23, P < 0.001), suggesting a contractility-dependent velocity of propagation.

Taken together, these data suggest that indices of ventriculo-arterial efficacy, cardiac afterload, and peripheral arterial stiffness while all are affected by CV disease, each cannot be used to predict one another, and all provide independent prognostic information in disease.
Abstract P608: Obesity Induced Hypertension Exacerbated Through Upregulation Of Renal Naci Cotransporter, Reversed By Increasing Pgc-1α-ho-1 Gene Expression To Restore Mitochondrial Function
Authors: Bamshad David , Cao Jian , Schragenheim Joseph , Stier Charles T , Abraham Nader G 
Published: June 2021

Introduction:
Hypertension caused by chronic obesity as a result of high calorie food intake or in leptin receptor deficient db/db mice may be linked to mitochondrial dysfunction. Previously we and others have shown that an epoxyeicosatrienoic acid agonist (EET-A), reduced adiposity and ROS resulting in normalization of BP by unknown mechanisms. We hypothesize that EET-A will attenuate BP by restoring mitochondrial function through increasing the PGC-1α-HO-1 axis and increasing urinary sodium excretion by downregulating NCC channels.

Methods:
Db/db mice at 16-wks of age were divided into 3 treatment groups and for an additional 16-wks received: A) control, B) EET-A 1.5mg/100g BW i.p. 2x/week and C) EET-A and lentiviral (Ln)- PGC-1α shRNA (to suppress PGC-1α protein). Oxygen consumption (VO
2
), visceral fat and blood glucose were determined. Additionally, renal tissues were harvested to measure the type 2 Na-K-Cl cotransporters (NKCC2), epithelial Na channels- (ENaC), NaCl cotransporters (NCC), PGC-1α, HO-1, insulin receptors, and mitochondrial biogenesis markers.

Results:
At the conclusion of 32 weeks:

Group A, developed hypertension and presented with decreased urinary Na excretion, decreased VO
2
, decreased downstream PGC-1α signaling, and mitochondrial dysfunction. There were increased levels of NCCs but not of NKCC2s or ENaCs. Renal PGC-1α, HO-1, pAMPK, and mitochondrial fusion protein Mfn 1/2, and Opa1 were decreased, p<0.05.

Group B, exhibited restoration of renal levels of PGC-1α, HO-1, pAMPK, and mitochondrial biogenesis proteins Mfn 1/2 and Opa1. NCC expression was reduced and was associated with an increase in urinary Na excretion; (p<0.05).
The beneficial effect of EET-A observed in group B was suppressed in group C using Ln- PGC-1α shRNA which suppressed PGC-1α expression in renal tissue > 50% and was accompanied by the onset of even more severe suppression of urinary Na excretion than in Group A.

Conclusion:
Treatment of obese mice with EET-agonists leads to the recruitment of PGC-1α-HO-1 which enhances mitochondrial function and induces the downregulation of NCC channels and increased sodium excretion. EET may serve as a powerful therapeutic agent for the treatment of obesity induced hypertension.
Abstract 270: mTORC1 is Essential for Cardiomyocyte Homeostasis and Female Sex Related Cardioprotection
Authors: Guergen Dennis B , Beiner Nelli , Catar Rusan , Mielke Thorsten , Busch Hauke , Boerries Melanie , Dengjel Joern , Brink Marijke , Grahammer Florian , Huber Tobias B , Kusch Angelika , Dragun Duska 
Published: June 2021

Mechanistic target of rapamycin (mTOR) kinase operates in two functionally and structurally distinct protein complexes mTORC1 and mTORC2. Raptor is a defining mTORC1 protein critical for cardiac growth in response to stress. Our previous studies showed sexual dimorphism in LV remodeling of normotensive DOCA-salt mice, where females retain normal LV architecture and males developed maladaptive LV hypertrophy. We hypothesized involvement of mTORC1 in regulation of this sexual dimorphism and induced cardiomyocyte (CM) specific raptor deletion (ra-cKO) in our model of normotensive mineralocorticoid challenge. Deletion of raptor in CM lead to decreased mTORC1 downstream target activation of 4E-BP1 and strong compensatory hyperactivation of mTORC2 target AKT in both sexes.
In contrast to healthy wild type females, female DOCA-salt ra-cKO mice developed severe dilative phenotype with tremendous interstitial fibrosis (15fold increase vs. WT) of similar extent as males. Functionally, ra-cKO females had stronger decrease in EF (51 vs. 26%) than ra-cKO males (50 vs. 32%). Greater loss of sarcomeric ultrastructure and mitochondrial morphology in transmission electron microscopy was observed in male ra-cKOs. Analysis of label-free proteomics and transcriptomics approaches revealed genotype and sex-specific clustering and identified significantly regulated single proteins or gene sets. Concordant to observed ultrastructural changes, down-regulation of genes for Z-disc, t-tubule and sarcomere organization was prominent in male hearts, whereas females displayed up-regulation of gene sets for inflammation, angiogenesis and mitochondrial function.
Intact mTORC1 is indispensible for CM homeostasis of both sexes as reflected by development of severe phenotype even by a mild stressor in our model. More specific, female sex-specific cardioprotection is completely lost in absence of mTORC1. Despite similar phenotypic changes in both sexes, sex-specific differences in regulation of gene expression points toward sex-specific functional selectivity of mTORC1 in regulation of CM homeostasis. Given the broad indications for mTOR inhibitors and especially used high concentration in oncology, one could anticipate adverse cardiac side effects.
Abstract 50: DNA Methylation Signatures in Blood Distinguish Patients at Risk of Post-operative Atrial Fibrillation Following Cardiopulmonary Bypass
Authors: Monte Emma , Fischer Matthew A , Wisniewski Nicholas , Scovotti Jennifer , Howard-Quijano Kimberly , Kimball Todd , Hofer Ira , Wang Yibin , Pellegrini Matteo , Mahajan Aman , Vondriska Thomas 
Published: June 2021

Post-operative atrial fibrillation (AF) is the most common complication for cardiac surgery, with an incidence of 27-40% and is associated with increased 30-day mortality. Several models exist to predict post-op AF using clinical data, however none have been widely implemented, and all have room for improvement (average area under ROC curve is ~0.70). We tested whether DNA methylation, assayed from pre-operative blood draws, could add predictive value.
We enrolled a cohort of 55 adult patients undergoing cardiopulmonary bypass, with a 36% incidence of post-op AF. Reduced representative bisulfite sequencing provided single-base resolution for an average of 3 million CpGs with 10x sequencing coverage per sample. We first tested whether these CpGs were correlated with AF. Of the 809,569 CpGs that passed thresholds in all samples, 16,774 were significantly correlated with AF (corrected p-value <=0.01 using methylKit in R). We used the R package Glmnet to perform logistic regression on the 16,774 CpGs in 40 of the 55 patients using 5-fold cross validation to build a model and selecting the 381 CpGs which were included in 95% of the 5,000 models. Our assumption being that these 381 CpGs were predictive no matter the subset of patients selected in the randomly generated groups employed during cross validation. Using these 381 CpGs as inputs we built a final model that ultimately incorporated 51 CpGs. None of the 51 CpGs were significantly correlated with age (corrected p-value <=0.01), ruling out age as a confounding factor. We tested our model on the remaining 15 patients. Our model correctly predicted 5 of the 5 AF cases, and 9 of the 10 AF-free cases. Validation of our model in a larger cohort is ongoing, and future directions will incorporate our epigenomic signature with readily available clinical risk factors to further optimize the model.
Abstract 81: Absence of Macrophage microRNA-155 Affects Cardiac Metabolic and Inflammatory Remodeling Following Diabesotension and Protects Against Development of Heart Failure
Authors: Rech Monika , Wouters Kristiaan , Lecomte Julie , Bitsch Nicole , Heymans Stephane , van Bilsen Marc , Schroen Blanche 
Published: June 2021

The co-existence of diabetes, obesity, hypertension - diabesotension - is a major burden in western societies and often culminates into heart failure (HF). Diabesotension is associated with a chronic status of low-grade inflammation, involving macrophage infiltration into adipose and cardiac tissues, local activation of inflammatory pathways and concomitant insulin-resistance. We showed that the microRNA miR-155 is a powerful modulator of hypertension-induced cardiac inflammation and HF. Here we hypothesize that absence of miR-155 in macrophages protects against adipose tissue and cardiac inflammation during diabesotension and thereby prevents insulin resistance and cardiac dysfunction.
Male C57Bl/6J mice received a bone marrow transplantation of miR-155 WT (n=30) or miR-155 KO (n=30) donors with same genetic background. High fat (HFD; n=22/gr) or chow diet (n=8/gr) was given for 20 weeks. An HFD subgroup was subjected to pressure overload by angiotensin II (1.5mg/kg/d) for 4 weeks (HFD+PO; n=12/gr). Glycaemia, glucose tolerance test, cardiac MRI were performed after 16 and/or 20 weeks. Organs were stored for histology, RNA and FACS analysis.
After 16 weeks of HFD, body weight (+11% WT; +14% KO; p<0.05) and glycaemia (+16% WT; +18% KO; p<0.05) increased compared to chow diet animals in both genotypes, confirming the status of obesity and pre-diabetes. Absence of macrophage miR-155 protected HFD-mice from glucose intolerance (at T15: WT 25.08; KO 18.15 mmol/L; p<0.05), suggesting that adipose tissue macrophages might contribute to insulin resistance. In addition, HFD+PO caused mild systolic dysfunction in WT but not in miR-155 KO animals compared to control (ejection fraction: -19% WT, p<0.05; -7% KO, n.s.). While cardiac interstitial fibrosis was not affected by absence of macrophage miR-155, cardiac CD45-positive leukocyte infiltration upon HFD+PO was abrogated. HFD+PO-induced cardiac expression of two PPAR-dependent metabolic genes, angiopoietin-like 4 and uncoupling protein 3, is repressed in the absence of macrophage miR-155.
Our data suggest that macrophage miR-155 plays a role in the development of insulin resistance and cardiac dysfunction, possibly by affecting local immune cell function and metabolic programming.
Abstract TMP69: Pre-Mixing IV TPA for Acute Ischemic Stroke Treatment: Impact on Door to Needle Times, SICH Rates, Patient Selection, and Amount of Drug Wasted With Associated Costs
Authors: Sessa Joy , Sherman Veronica , Doctor Norman , Rogers Bradley , Ham Jared , Vinitskaya Irina , DiMartini Gina , Mehta Brijesh 
Published: June 2021

Acute ischemic stroke (AIS) is a significant concern in the United States with 795,000 people suffering strokes annually (Benjamin, 2019). The irreversible effects of a minute of acute brain ischemia is significant—1.9 million neurons lost (Saver, 2006). Thrombolytic therapy has been a key component of stroke treatment with McTaggart, (2015) recommending a 30 minute door to needle (DTN) goal. Target Stroke II (2017) recommends pre-mixing tPA. However, little is known about the safety and efficacy of this practice.

This study retrospectively reviewed 211 AIS patients treated at 2 stroke centers to measure the efficacy & safety of a standardized pre-mix criteria for tPA administration. Variables included DTN times, SICH rates, rate of tPA vials wasted & reimbursed, and patient selection (acute infarct on imaging). Study inclusion criteria: adults (≥ 18 years) without contraindications to tPA, baseline NIHSS
>
4, onset of focal neurological deficits < 4.5 hrs, not on anticoagulants, & initial SBP
<
185.

Treatment group patients (n=107) received pre-mixed tPA (mixed prior to resulted NCCT). Retrospective comparison group patients (n=104) met the same inclusion parameters as the treatment group.
Study results: median DTN time for the comparison group: 30 min (StD=19.6); treatment group: 20 min (StD=13.5) (p = 0.00002), a 33% decrease in DTN time. SICH rates for the comparison group: 1.4%; treatment group: 0.8%. tPA vials wasted: comparison group: 13%, treatment group: 22%, a 9% increase in vial wastage when pre-mixing tPA. The percentage of wasted vials reimbursed by the manufacturer was 100% for both groups. Patient selection accuracy was 66% for the comparison group and 51% for the treatment group, a 15% accuracy decrease for the pre-mix group.
In conclusion, implementing a pre-mix criteria for tPA decreased DTN times significantly without increasing SICH rates. Use of a pre-mix criteria did not demonstrate improved patient selection, evidenced by the lower rate of acute infarcts on imaging in pre-mixed patients. Although the drug wastage increased in the treatment group, reimbursement rates remained at 100%. Utilizing a standardized pre-mix criteria for tPA is a safe, effective way to decrease door to needle times without compromising financial stability.