bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of the Adenosine Pathway to Potentiate Cancer Immunotherapy: Potential for Combinatorial Approaches
Authors: Thompson Elizabeth A. , Powell Jonathan D.
Published: September 2020

Cancer immunotherapy has revolutionized the way that we think about treating cancer. Although checkpoint blockade therapy, including anti-PD-1/PD-L1 and anti-CTLA-4, has shown remarkable success, the responses are limited to only a subset of patients. This discrepancy highlights the many overlapping avenues for immune evasion or suppression that can be employed by a tumor. One such mechanism of immunosuppression is adenosinergic signaling within the tumor microenvironment. We provide an overview of the current status of clinical trials targeting the adenosine pathway, including CD73, CD39, and adenosine receptors. Additionally, we highlight several avenues that may be explored to further potentiate responses in the clinic by combining adenosine-targeting agents to target multiple arms of the pathway or by using conventional immunotherapy agents.
Clostridioides difficile Spore Formation and Germination: New Insights and Opportunities for Intervention
Authors: Shen Aimee
Published: September 2020

Spore formation and germination are essential for the bacterial pathogen Clostridioides difficile to transmit infection. Despite the importance of these developmental processes to the infection cycle of C. difficile, the molecular mechanisms underlying how this obligate anaerobe forms infectious spores and how these spores germinate to initiate infection were largely unknown until recently. Work in the last decade has revealed that C. difficile uses a distinct mechanism for sensing and transducing germinant signals relative to previously characterized spore formers. The C. difficile spore assembly pathway also exhibits notable differences relative to Bacillus spp., where spore formation has been more extensively studied. For both these processes, factors that are conserved only in C. difficile or the related Peptostreptococcaceae family are employed, and even highly conserved spore proteins can have differential functions or requirements in C. difficile compared to other spore formers. This review summarizes our current understanding of the mechanisms controlling C. difficile spore formation and germination and describes strategies for inhibiting these processes to prevent C. difficile infection and disease recurrence.
Evaluating the Sustainable Growth of Small- and Medium-Sized Construction Enterprises Using the Multicriteria Decision-Making Method
Authors: Wang Wenbao , Lin Wenhe , Dai Xinyi , Yan Yingzheng , Xuan Aiping
Published: September 2020

Sustainable growth plays an important role in the development of small- and medium-sized construction enterprises and has increasingly become a hot topic in the political and academic arenas. Growth is an important indicator for measuring the operational status and development potential of listed enterprises and is of great significance for studying sustainable growth. Considering the development of small- and medium-sized construction enterprises, a growth evaluation index system that includes growth resources, growth ability, and growth innovation was established, and a growth evaluation model was constructed using the improved entropy-VIKOR algorithm. Evaluation and analysis of the sustainable growth of small- and medium-sized construction enterprises listed before the end of 2012 were conducted. The parameter sensitivity, weight sensitivity, and effectiveness of the evaluation model were analysed. The empirical results showed that compared with the TOPSIS method and the traditional VIKOR method, the multicriteria evaluation model for the sustainable growth of small- and medium-sized construction enterprises proposed in this paper had better stability and effectiveness. Finally, the key influencing factors, growth ranking, and dynamic growth types of small- and medium-sized construction enterprises were analysed, and targeted suggestions were proposed. The research results can provide a reference for managers, investors, and regulators.
Transcriptional Expressions of CXCL9/10/12/13 as Prognosis Factors in Breast Cancer
Authors: Li Yan , Liang Mingqiang , Lin Yuxiang , Lv Jinxing , Chen Minyan , Zhou Peng , Fu Fangmeng , Wang Chuan
Published: September 2020

CXCLs play critical roles in antitumor immunity by activating tumor-specific immune responses and stimulating tumor proliferation, thus affecting patient outcomes. However, the expression and prognostic values of CXCLs in breast cancer have not been well clarified. The aim of this study was to investigate the impact of CXCLs transcriptional expression on breast cancer patients. Oncomine database, GEPIA (Gene Expression Profiling Interactive Analysis), UALCAN, Kaplan–Meier Plotter, TIMER (Tumor Immune Estimation Resource), and DAVID were used in our study. The transcriptional levels of CXCL9/10/11/13 in breast cancer tissues were significantly elevated while the transcriptional levels of CXCL1/2/3/12 were decreased based on intersections of Oncomine database and GEPIA. Among them, breast cancer patients with high transcriptional levels of CXCL2/9/10/12/13 and low transcriptional level of CXCL3 were associated with a better prognosis. We also found that most of CXCLs expressions are significantly correlated with known prognostic factors, such as patient’s age, major subclasses, individual cancer stages, and nodal metastasis status. In addition, the expression of CXCL9/10/12/13 was also indicated to be correlated with the infiltration of six types of immune cells (B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and dendritic cells). The functions of differentially expressed CXCLs are primarily related to the immune response and cytokine-cytokine receptor interactions. Our results may provide novel evidence of new prognostic or predictive biomarkers for breast cancer patients.
Efficient Task Offloading for 802.11p-Based Cloud-Aware Mobile Fog Computing System in Vehicular Networks
Authors: Wu Qiong , Ge Hongmei , Fan Qiang , Yin Wei , Chang Bo , Wu Guilu
Published: September 2020

Various emerging vehicular applications such as autonomous driving and safety early warning are used to improve the traffic safety and ensure passenger comfort. The completion of these applications necessitates significant computational resources to perform enormous latency-sensitive/nonlatency-sensitive and computation-intensive tasks. It is hard for vehicles to satisfy the computation requirements of these applications due to the limit computational capability of the on-board computer. To solve the problem, many works have proposed some efficient task offloading schemes in computing paradigms such as mobile fog computing (MFC) for the vehicular network. In the MFC, vehicles adopt the IEEE 802.11p protocol to transmit tasks. According to the IEEE 802.11p, tasks can be divided into high priority and low priority according to the delay requirements. However, no existing task offloading work takes into account the different priorities of tasks transmitted by different access categories (ACs) of IEEE 802.11p. In this paper, we propose an efficient task offloading strategy to maximize the long-term expected system reward in terms of reducing the executing time of tasks. Specifically, we jointly consider the impact of priorities of tasks transmitted by different ACs, mobility of vehicles, and the arrival/departure of computing tasks, and then transform the offloading problem into a semi-Markov decision process (SMDP) model. Afterwards, we adopt the relative value iterative algorithm to solve the SMDP model to find the optimal task offloading strategy. Finally, we evaluate the performance of the proposed scheme by extensive experiments. Numerical results indicate that the proposed offloading strategy performs well compared to the greedy algorithm.
ATP-Dependent Chromatin Remodeling Complex in the Lineage Specification of Mesenchymal Stem Cells
Authors: Du Wen , Guo Daimo , Du Wei
Published: September 2020

Mesenchymal stem cells (MSCs) present in multiple tissues can self-renew and differentiate into multiple lineages including the bone, cartilage, muscle, cardiac tissue, and connective tissue. Key events, including cell proliferation, lineage commitment, and MSC differentiation, are ensured by precise gene expression regulation. ATP-dependent chromatin alteration is one form of epigenetic modifications that can regulate the transcriptional level of specific genes by utilizing the energy from ATP hydrolysis to reorganize chromatin structure. ATP-dependent chromatin remodeling complexes consist of a variety of subunits that together perform multiple functions in self-renewal and lineage specification. This review highlights the important role of ATP-dependent chromatin remodeling complexes and their different subunits in modulating MSC fate determination and discusses the proposed mechanisms by which ATP-dependent chromatin remodelers function.
3D Global Weak-Form Mesh-Free Method for Acoustic Attenuation Analysis of Expansion Chambers
Authors: Fang Chun-Hui , Liu Cheng-Yang , Fang Zhi
Published: September 2020

In order to avoid the dependence of mesh method on grids, a 3D global weak-form mesh-free method (MFM) is applied to study the three-dimensional acoustic characteristics of silencers. For the expansion chamber silencers, the 3D acoustic modes are extracted and the transmission loss results are computed by using the 3D global weak-form (MFM), which is based on the radial basis function point interpolation method (RPIM) for calculating the shape functions and Galerkin method for discretizing the system equation. The first 15 order 3D acoustic modes and TL results of a special expansion chamber silencer are presented to validate the computational accuracy of the proposed technique, and the relative errors are controlled within 0.5% by comparing with the 3D finite element method (FEF) calculations. Additionally, the effects of axial modes on the acoustic characteristics are investigated, and the pass through frequencies can be eliminated to enhance the acoustic attenuation performance by locating the side branch outlet on the nodal lines of axial modes.
Different Innate Immune Responses in BALB/c and C57BL/6 Strains following Corneal Transplantation
Authors: Bleul Tim , Zhuang Xinyu , Hildebrand Antonia , Lange Clemens , Böhringer Daniel , Schlunck Günther , Reinhard Thomas , Lapp Thabo
Published: September 2020

Purpose: To investigate immunological differences and the role of CD38+/F4/80 + M1 macrophages in C57BL/6J- and BALB/c-recipient mouse corneal transplantation models. Methods: Allogeneic transplantation was performed crosswise in BALB/c mice and C57BL/6J mice; syngeneic transplantation was performed in both strains. Anterior chamber depth (ACD) was measured before and central corneal thickness (CCT) was measured both before and after transplantation. In vivo graft rejection was monitored using anterior eye segment optical coherence tomography (ASOCT) evaluating the CCT and grading of corneal oedema using a well-established clinical score (CS). Histology of corneal grafts was performed 18 or 21 days after surgery. Immunohistochemistry with anti-F4/80 antibody and anti-CD38 antibody was used for detecting M1 macrophages within the grafts. Results: High CS and CCT values after allogeneic transplantation persisted in both BALB/c (n = 18) and C57BL/6J recipients (n = 20). After syngeneic transplantation, CS and CCT values increased in both models in the early phase after surgery due to the surgical trauma. Surprisingly, in the syngeneic C57BL/6J model, high CCT values persisted. Furthermore, anterior synechiae developed in C57BL/6 recipients after both syngeneic and allogeneic transplantation, whereas BALB/c recipients showed almost no synechiae – even though C57/BL6J animals tended to have a deeper anterior chamber (281 ± 11 pixels [mean ± SD]) compared with BALB/c animals of the same age (270 ± 9 pixels [mean ± SD]). Immunohistochemistry revealed numerous CD38+/F4/80 + M1 macrophages in grafts of C57BL/6J recipients following both syngeneic and allogeneic transplantation. However, in BALB/c-recipient mice only sparse M1 macrophages were detectable (CD38 + M1 macrophages relative to all F4/80 + cells: 75 vs. 17% [after allogeneic transplantation] and 66 vs. 17% [after syngeneic transplantation]; p < 0.05). Conclusions: Allogeneic corneal transplants are rejected in BALB/c as well as C57BL/6J mice, but tissue alterations with anterior synechiae are more pronounced in C57BL/6J recipients. Following syngeneic transplantation, C57BL/6J-recipient animals show a persistent graft swelling with increased numbers of CD38+/F4/80 + M1 macrophages in grafted tissue, in contrast to the common model using BALB/c-recipient mice. Our data strongly suggest that strain-dependent differences convey different innate immune responses in BALB/c and C57BL/6J strains. Accordingly, in murine keratoplasty experiments, the mouse line of both donor and recipient animals must be carefully considered. C57BL/6J-recipient mice might be particularly suited to study corneal graft rejection in a clinical setting considered “high risk.”
Presence of Parvovirus B19 but Not Herpesvirus Genome in Acute Skin Rash after Allogeneic Stem Cell Transplantation Correlates with Outcome
Authors: Weber Thomas , Schmidberger Andreas , Ligeti Kinga , Bauer Marcus , Rosenwald Andreas , Müller Lutz P.
Published: September 2020

Introduction: Skin rash is a first symptom of acute graft-versus-host disease (GvHD) after allogeneic stem cell transplantation (ASCT) but can also be caused by viruses. The relevance of virus DNA analyses in skin rash for diagnosis and clinical outcome is unknown. Objectives: To record the frequencies of detection of herpes and parvovirus B19 (ParvoB19) DNA in skin rash within 100 days after ASCT and to analyze their relevance for diagnosis, clinical course, and non-relapse mortality (NRM). Methods: We retrospectively identified 55 patients with virus DNA analysis for CMV, EBV, HHV6, HHV8, HSV, VZV, or ParvoB19. We assessed the rate of virus DNA detection and studied associations with histological diagnosis, virus DNA from concomitantly analyzed blood, clinical presentation, exanthema treatment, and NRM. Results: CMV, EBV, HHV6, HHV8, HSV, VZV and ParvoB19 DNA were detected in 12.5, 11.8, 10, 0, 0, 2.9, and 26.7% of exanthemas. Histopathological diagnosis was not associated with virus polymerase chain reaction (PCR) results. Detection of CMV, EBV, or HHV6 DNA but not ParvoB19 in skin and blood was associated with PCR results (p = 0.016; p < 0.001; p = 0.067; p = n.a.). Detection of CMV, EBV, HHV6, or ParvoB19 DNA in the skin was not significantly associated with patient, ASCT, or GvHD characteristics. Detection of ParvoB19 but not herpes virus DNA was associated with less immunosuppressive treatment (p = 0.015) and lower NRM (p = 0.041). In multivariate analyses, detection of ParvoB19 was associated with a lower NRM. Conclusions: Detection of ParvoB19 DNA in exanthema after ASCT might be associated with lower NRM.
Concomitant Primary Hyperparathyroidism in Patients with Multiple Myeloma: A Possible Link?
Authors: Notarfranchi Laura , Marchica Valentina , Dalla Palma Benedetta , Pelagatti Laura , Burroughs-Garcia Jessica , Pedrazzoni Mario , Ruffini Livia , Cetani Filomena , Marcocci Claudio , Giuliani Nicola
Published: September 2020

Hypercalcemia is a significant feature of patients with active multiple myeloma (MM) with extensive bone disease. Among the causes of non-neoplastic hypercalcemia, primary hyperparathyroidism (PHPT) is one of the most common, leading to osteoporosis and bone fractures. Interestingly, some preclinical data indicate that high secretion of parathyroid hormone (PTH) may have a negative impact on bone disease and MM progression. However, concomitant diagnosis of MM and PHPT has rarely been described. Here, we present 4 cases of patients with active MM and hypercalcemia with high or inappropriately normal PTH levels. Interestingly, CD138+ cells from these 4 MM patients lack PTH receptor 1 and PTH-related peptide expressions, indicating that PTH could have a paracrine rather than a direct pro-tumoral effect. Moreover, these cases suggest that the concomitant diagnosis of MM and PHTP may not be so rare and should be considered for the clinical management of MM patients with hypercalcemia.
The Association of Introducing a Faecal Calprotectin Testing Pathway for Suspected Inflammatory Bowel Disease in Primary Care and Time to Diagnosis or Treatment
Authors: Hicks Amy , Hamlin P. John , Selinger Christian P.
Published: September 2020

Background: Primary care faecal calprotectin (FC) was introduced in Leeds in 2014 to distinguish inflammatory bowel disease (IBD) from irritable bowel syndrome and with the hope that it may reduce time to IBD diagnosis and treatment. This study examines the association of FC with referral routes, time to diagnosis, and time to treatment. Methods: All patients newly referred to IBD clinics in 2013 and 2016 were studied. Data on referral routes and dates, FC, date of first treatment, and proxy outcomes for disease severity were collected. Results: In 248 patients, there were no differences between 2013 and 2016 cohorts regarding baseline data and disease severity. The number of direct referrals to gastroenterology rose from 3% (2013) to 17% (2016), whilst 10% were diagnosed during emergency admissions. Referrals via suspected cancer pathways remained high (38% in 2013, 28% in 2016), whilst many had initial investigations at independent centres (16% in 2013, 24% in 2016). Time from referral to diagnosis was similar between 2013 (0.77 month) and 2016 (1.10 months, p = 0.2). A total of 48 (33.3%) patients had FC checked prior to referral, and 37.5% of these were referred directly to gastroenterology. Time from diagnosis to treatment reduced from 1.37 months (2013) to 0.72 month (2016, p = 0.01). Conclusion: Patients present via a multitude of referral pathways, but FC was associated with increased direct referrals to gastroenterology. We found a variation in time to diagnosis and treatment depending on referral routes. Further work is required to ensure patients with suspected IBD get referred to IBD services in a timely manner.
Age and Diastolic Blood Pressure Play an Important Role in the Progression of White Matter Lesions: A Meta-Analysis
Authors: Zhang Dong , Tang Yi , Ge Jian , Liu Yumeng , Jin Jing , He Mingli
Published: September 2020

Introduction: White matter lesions (WMLs) are currently considered as a cerebral microvascular disease, and hypertension is considered as its most important risk factor. This study analysis systematically evaluated the effects of systolic blood pressure (SBP) and diastolic blood pressure (DBP) on the progression of WMLs. Methods: We searched the MeSH terms: “white matter,” “blood pressure,” “hypertension,” “Leukoencephalopathy,” and “leukoaraiosis” on PubMed and Cochrane from January 2000 to August 2019. A total of 12 closely related literature were included for research. Results: The results of the meta-analysis showed that the increase of both SBP and DBP can promote the progression of WMLs (respectively, odds ratio [OR] = 2.90, 95% confidence interval [CI] 2.86–2.94; OR = 3.13, 95% CI 3.03–3.23). Subgroup analysis found that patients with hypertension aged younger than 70 years are at a greater risk of WML progression when their DBP increased (OR = 4.69, 95% CI 3.31–6.65). Conclusion: The relationship between DBP and WMLs is closer than that of SBP. Also, the risk of WML progression in patients aged under 70 years was higher than that in patients aged over 70 years. Furthermore, when DBP is elevated in patients younger than 70 years of age, the risk of WML progression may be higher. Therefore, it is expected that more researchers will attach importance to the change in DBP and identify the range of blood pressure and strategies that control DBP, thus contributing to delay the progression of WMLs.
Abstract 14: Glyburide Treatment Improves Aortic Arch Pulse Wave Velocity In A Murine Model Of Alzheimer’S Disease
Authors: Cruz Diaz Nildris , Day Stephen M. , Snipes J. Andrew , Diz Debra I. , Macauley Shannon L.
Published: September 2020

Brain hyperexcitability is a defining feature of AD, where aberrant neuronal activity may be both a cause and consequence of the pathology. Identifying novel targets to modulate cellular excitability is an important treatment strategy. We showed that inward rectifying, ATP-sensitive potassium (K
ATP
) channels regulate excitability at the neurovascular unit to impact production and aggregation of amyloid-beta (Aβ), a hallmark of AD. Sulfonylureas such as glyburide (GLY) are antidiabetic medications that inhibit K
ATP
channels. We demonstrated that systemic treatment with GLY decreased Aβ production and plaque pathology by modulating vascular K
ATP
channel activity and neuronal excitability. Here, we investigated whether GLY alters cardiac function or pulse wave velocity (PWV) in APPswe, PSEN1dE9 (APP/PS1) mice, a model of Aβ overexpression.Female APP/PS1 mice at 8 months of age with established amyloid plaque pathology were treated for 1 month with GLY (subcutaneous, slow release 2.5mg pellet, ~30μg/GLY/day) or Placebo (n = 5, each group). After 1 month, transthoracic echocardiography was performed (Vevo 2100 LAZR, FUJIFILM/VisualSonics, Inc.; Toronto, Ca) with a 30 MHz linear array transducer. There were no differences in Placebo vs. GLY mice for heart rate, cardiac output, E/e’ ratio, ejection fraction, and fractional shortening. Arterial stiffness, measured as PWV in the aortic arch [Distance (D)/Time (T); D = mm from ascending to descending aorta and T = (R to ascending aorta foot) - (R to descending aorta foot) in ms], was lower in GLY treated mice (5 ± 1 vs. Placebo 9 ± 1 mm/ms; p < 0.04). Wild Type females at 9 months of age (n = 5) reveal similar cardiac function values as Placebo and GLY treated APP/PS1 mice, with a trend for lower PWV (6 ± 1 mm/ms) than the APP/PSI Placebo group. Thus, short-term treatment with GLY does not impact overall cardiac function. Moreover, improved vascular stiffness in GLY-treated APP/PS1 mice may contribute to improved neurovascular coupling independent of changes in Aβ aggregation, since arterial stiffness is associated with increased cerebrovascular pathology in clinical studies. SLM: Bright Focus Fdn, NIA K01AG050719, Donors Cure New Vision; SMD: T32AA007565; Hypertension & Vasc Res Ctr; NCATS UL1TR001420 (Vevo Core)
Abstract 15: The Role Of Opioid Receptors In Podocytes In The Development Of Hypertension In Dahl Salt-sensitive Rats
Authors: Golosova Daria , Palygin Oleg , Levchenko Vladislav , Klemens Christine A , El-Meanawy Ashraf , Staruschenko Alexander
Published: September 2020

The rise in opioid use underscores the importance to better understand the direct and indirect effects of opioids on renal function and blood pressure. Although opioid use is associated with predictors of cardiovascular diseases, these drugs are common analgesics for hypertensive patients. We hypothesize that stimulation of opioid receptors (ORs) leads to elevated intracellular calcium level in podocytes ultimately leading to cell apoptosis, development of albuminuria and consequent progression of hypertension. Live calcium imaging experiments on freshly isolated glomeruli from rat and human kidneys, as well as human immortalized podocyte cell line, was performed to test the effect of specific ORs agonists. Following experiments assessed the effect of opioid signaling on the development of hypertension and kidney function in Dahl salt-sensitive (SS) rats, which were fed a 0.4% (LS) or 8% (HS) NaCl diets for 14 days with or without a daily
i.v.
bolus infusion of BRL52537, a potent and selective kappa-OR agonist. Stimulation of kappa-ORs, but not mu-ORs or delta-ORs, mediated calcium influx in podocytes through activation of TRPC6 channels. The effect of BRL52537 was completely abolished when we used the 0 mM calcium media or when a TRPC6 channel inhibitor (SAR7334) was applied. Triggering the kappa-OR/TRPC6 pathway induced podocyte cell shape changes via actin cytoskeleton remodeling.
In vivo
studies revealed that rats chronically treated with BRL52537 exhibited augmented blood pressure (MAP was 179 ± 15 vs. 151 ± 11 mmHg), albuminuria, and elevation in podocyte calcium. Western blot analysis revealed elevated levels of nephrin in urine samples and pro-caspase-3 in renal cortex. Moreover, TRPC6 expression was elevated under hypertensive conditions and further promoted pathological increase in calcium influx in response to kappa-OR stimulation. Summarizing the data, the opioid-induced increase in the calcium flux in podocytes is expected to contribute to kidney injury leading to progression of salt-induced hypertension. These data demonstrate that the kappa-OR/TRPC6 signaling pathway directly influences podocyte calcium handling, provoking the development of kidney injury in the opioid treated hypertensive cohort.
Abstract 16: Progressive Alterations Of Mitochondrial Bioenergetics In The Kidney During The Development Of Salt-induced Hypertension
Authors: Tomar Namrata , Kandel Sunil M , Zhang Xiao , Zheleznova Nadezhda , Cowley Allen W , Dash Ranjan K
Published: September 2020

Hypertension is a complex disease and a leading cause of morbidity and mortality globally. Although oxidative stress and mitochondrial dysfunction have been found in the kidney in various models of hypertension, progressive alteration of mitochondrial oxidative phosphorylation (OxPhos) in the kidney during the development of salt-sensitive hypertension has not been characterized. The present study determined changes of OxPhos in kidneys of Dahl salt-sensitive (SS) rats before (0.4% NaCl diet; LS) and after switching to a high salt diet (4.0% NaCl; HS) during the development of hypertension. Mitochondria were isolated from the outer medulla (OM) and cortex of the kidney of SS rats fed a LS diet since weaning and studied at days 3, 7, 14 & 21 of a HS diet feeding. Oxygen consumption rates (OCR) were measured in mitochondria energized with pyruvate + malate as substrates for three different respiratory states using an Oroboros Oxygraph-2k Instrument. This includes i) leak state (in the absence of ADP), ii) ADP-stimulated state, and iii) uncoupled state (in the presence of an uncoupler FCCP). A biphasic pattern of ADP-stimulated OCR with progressive uncoupling was observed in both the renal OM and cortex. Mitochondrial efficiency for ATP synthesis was increased in the early phases of hypertension (3 & 7 days) but was severely compromised in the established phases of hypertension (14 & 21 days). This decreased mitochondrial efficiency was associated with uncoupling of OxPhos and high levels of oxidative stress which we hypothesized were due to mitochondrial ROS stimulation of membrane NOXs. To test this, experiments were performed in SS rats with double knock out (DKO) of the cytosolic subunit of NOX2 (p67
phox
) and NOX4 (SS
p67phox-/-/Nox4-/-
). DKO SS rats were fed a HS diet and OCR of renal cortical and OM mitochondria was determined at days 7 and 14. In contrast to SS rats, the DKO SS rats fed a HS diet showed no significant differences in mitochondrial OCR in the cortex or OM, nor to a control group maintained on a LS diet. HS diet in SS rats initially increases the efficiency of renal cortical and medullary mitochondrial ATP production (days 1-7) followed by an enhanced ROS production with mitochondrial uncoupling and reduced efficiency of ATP production by the third week.
Abstract 18: Orexin Knockout BPH/2J Hypertensive Mice Still Have Elevated Basal Blood Pressure But Lower Stress Reactivity
Authors: Jackson Kristy L , Delerue Fabien , Kumar Natasha N , Ittner Lars M , Head Geoff , Carrive Pascal
Published: September 2020

Background:
Orexin is a neuropeptide which can influence a wide range of physiological functions including blood pressure, sympathetic nervous system activity and arousal. Orexinergic signalling is suggested to contribute to the neurogenic hypertension in BPH/2J mice, based on the hypotensive response to the dual orexin receptor (OxR) antagonist, Almorexant, which was not observed in normotensive BPN/3J controls. Orexin is also known to influence the cardiovascular response to certain stressors. Thus, it is possible that orexin may also contribute to the exaggerated pressor response to stress that is reported in BPH/2J mice.

Objective:
To determine the contribution of orexin to the hypertension and exaggerated stress reactivity in BPH/2J wild type mice by comparing with BPH/2J Orexin homozygous knock out (BPH OxKO) mice.

Methods:
BPH OxKO (n=9) and BPH wild type (BPH WT) mice (n=6) were implanted with radiotelemetry probes to measure blood pressure (BP), heart rate (HR) and locomotor activity across the 24hr period and in response to restraint and dirty cage swap stressors. The BP responses to ganglion blocker pentolinium (5mg/kg, i.p.), was also measured.

Results:
There was no difference in average mean arterial pressure (MAP) over the 24hr period in BPH OxKO mice (131±1 vs 133±1mmHg,
P
=0.351), but there was a modest elevation in systolic AP (+4.5mmHg) exclusively during light period in BPH OxKO compared with BPH WT mice (145±2 vs 141±2mmHg,
P
<0.032). HR was comparable between strains (
P
=0.160) and locomotor activity tended to be lower in BPH OxKO compared with BPH WT mice during the dark period (1.1±0.2 vs 1.7±0.3 units respectively,
P
=0.051). The pressor response to dirty cage swap stress was lower in BPH OxKO compared with BPH WT mice (22±1 vs 28±1mmHg respectively,
P
<0.001), as was the response to restraint stress (36±1 vs 40±2mmHg respectively,
P
<0.05). The depressor response to ganglion blockade was comparable between strains (
P
=0.255).

Conclusion:
The present findings suggest that whilst orexin does not contribute to sympathetically mediated hypertension, it may play a role in the exaggerated cardiovascular response to stress and hyperactivity in BPH/2J mice.
Abstract 22: Formyl Peptide Receptor-1 Activation Is Crucial For The Cause Of Spontaneous Hypertension In Dahl Salt Sensitive Rats
Authors: Edwards Jonnelle M , Galla Sarah , Bearss Nicole R , Mell Blair , Cheng Xi , Saha Piu , Vijay-Kumar Matam , McCarthy Cam , Joe Bina , Wenceslau Camilla F
Published: September 2020

Mitochondria evolved from bacteria and use N-formylated peptides (NFPs) to synthetize protein. Bacterial and mitochondrial NFPs activate formyl peptide receptor 1 (FPR-1) and lead to vascular injury. We previously observed that Dahl Salt Sensitive rats (S) fed a low-salt (LS, 0.3% NaCl) diet presented spontaneous hypertension, vascular dysfunction, and overexpression of FPR-1 in arteries when compared to Dahl Salt Resistant (R) rats. High salt (HS, 2% NaCl) diet worsened these phenotypes in S rats. Interestingly, HS diet induced leaky gut and amoxicillin (AMO) treatment decreased BP in S-HS. Due to the dual sources of NFPs (microbiota and host mitochondria), we hypothesized that cell death-derived mitochondria and/or leaky gut-derived bacterial NFPs lead to FPR-1 activation, vascular injury and elevated BP in S rats independent of HS diet. For this, we used flow cytometry to measure cell necrosis and early and late apoptosis in kidney, bone marrow-derived macrophages and mesenteric resistance arteries (MRA) from male S and R rats (8-week old) on a LS diet. Zonulin, a biomarker for leaky gut, was measured in plasma. In another group, rats were treated with FPR-1 antagonist [Cyclosporin H (CsH), 0.3 mg/kg/day, osmotic mini-pump, 14 days], vehicle (VEH) or received water with AMO (5 mg/kg/day) for 21 days to deplete bacteria. BP was measured by telemetry and vascular function and structure were assessed in MRA. S rats presented increased kidney cell necrosis (R: 3.8±0.3 vs. S: 5.3±0.5* %). CsH decreased spontaneous elevation of BP [Diastolic: R+VEH: 77±2.7 vs. R+CsH: 81±1.2 vs. S+VEH: 126±3.0* vs. S+CsH:115±2.7
#
] and vascular hypercontractility [KCl (120mM): R+VEH: 9.4±1 vs. R+CsH: 10.2±0.4; S+VEH: 15.5±0.9* vs. S+CsH:11.7±0.8
#
mN; Phenylephrine (10μM): R+VEH: 9.3±1 vs. R+CsH: 9.7±1; S+VEH: 14.5±1*vs. S+CsH: 11.4±0.6
#
mN) in S-LS rats. AMO did not change vascular contraction or BP. Leaky gut was not observed in Dahl S-LS diet. In conclusion, FPR-1 can serve as a causative agent for the spontaneous elevation of BP and kidney-derived mitochondria, but not gut-derived microbiota, are the main source for NFPs.
Abstract 3: Preeclampsia-induced T Helper-associated Cytokine Imbalances Persist Postpartum
Authors: Scroggins Sabrina M , Gray Gabrielle , Berkey Cassandra , Gandley Robin E , Sutton Elizabeth F , Gemmel Mary , Brandt Debra S , Santillan Donna A , Pierce Gary L , Hubel Carl A , Catov Janet M , Santillan Mark K
Published: September 2020

Preeclampsia (PreE), a hypertensive disorder in pregnancy, contributes to long-term maternal cardiovascular disease risk. By 2025, it is estimated that more women than men will have hypertension (HTN), yet the mechanisms contributing to the development of HTN in women are less understood. One potential mechanism underlying HTN in women is a persistent imbalance of anti- and pro- inflammatory T
H
cells following PreE. Consistent with this, anti-inflammatory T helper (T
H
) cytokines are reduced and pro-inflammatory T
H
cytokines are increased during a PreE pregnancy.

De-identified and coded plasma samples and clinical data were obtained from the Magee-Women’s Research Institute & Foundation or the University of Iowa Maternal-Fetal Tissue Bank (IRB 201808705) from women 1-3 (N=93) or 8-10 (N=58) years (yrs) following a normotensive (CTL) or PreE-affected pregnancy. Postpartum (PP) HTN was defined as having stage 1 or higher HTN as designated in the updated 2017 ACC/AHA guidelines. Women with PreE had higher rates of HTN at 1-3 years and at 8-10 years PP, (24% vs. 5% and 65% vs. 17%, all p<0.05) compared to women with a normotensive pregnancy.

To determine if T
H
cells play a role in the future development of HTN, we investigated if the T
H
cytokine changes observed in PreE persist 1-3 yrs and 8-10 yrs PP. Cytokine concentrations were determined via ELISAs and normalized to total protein. Average cytokine concentrations are reported in pg/g. At 1-3 yrs PP, concentrations of anti-inflammatory cytokines IL-4 (47 vs. 6819, p<0.05), IL-10 (1204 vs. 15042, p<0.05) and TGFβ (8.2x10
5
vs. 4.4x10
6
, p<0.05) were reduced in women with a prior PreE pregnancy vs. women with a CTL pregnancy. At 8-10 yrs PP, pro-inflammatory IL-6 (86 vs. 18, p<0.05) and TNFα (298 vs. 53, p<0.05) were both significantly increased in women with prior PreE compared to women with a CTL pregnancy.

Here, we confirm women with a prior PreE pregnancy present with a higher prevalence of HTN early (1-3 yrs) and later (8-10 yrs) PP compared to women with a normotensive pregnancy. Further, we show an altered T
H
cytokine milieu persists following delivery in women with PreE. This pro-inflammatory milieu is associated with increased rates of HTN and thus, may underlie the future development of HTN in women with a history of PreE.
Abstract 4: Molecular Profile Of Peripheral Blood Mononuclear Cells In Hypertensive Adolescents With Target Organ Damage
Authors: MOHAMMED ARIF Fnu , Urbina Elaine M , Sadayappan Sakthivel , Becker Richard
Published: September 2020

Introduction:
Primary hypertension (PH) is a multifactorial disease influenced by genetic, epigenetic, and environmental factors. Despite the occurrence of PH-associated cardiovascular events in youth, the molecular mechanism(s) of target organ damage (TOD) are unknown.

Objectives:
(1) To identify an epigenetic signature and gene expression profiles in adolescents with low blood pressure (BP) and normal left ventricular mass (LVM) compared to those with high BP and high LVM; and (2) to determine novel gene targets and associated signaling pathways for future investigation and intervention.

Methods and Results:
A total of 397 participants (mean age 15.6 ±1.7 years, 59% male, 63% Caucasian) were enrolled across the distribution of BP. The average daytime ambulatory systolic BP recorded in healthy and hypertensive participants was 112 ±9.71 and 133 ±7.2 mmHg (
p<0.05
) respectively. Clinical measures revealed higher body mass index (26.8 ±7.02
vs
29.6 ±7.88 Kg/m
2
;
p<0.05
), and abnormal circulatory HDL (47.4 ±12.1
vs
43.4 ±11.7 mg/dL;
p<0.05
), glucose (87.8 ±7.98
vs
90.8 ±8.17 mg/dL;
p<0.05
), insulin (17.8 ±14.3
vs
23.7 ±19 μIU/dL;
p<0.05
), creatinine (0.718 ±0.13
vs
0.727 ±0.17 mg/dL;
p<0.05
), uric acid (5.4 ±1.63
vs
6.04 ±1.52 mg/dL;
p<0.05
), CRP (1.35 ±1.8
vs
1.92 ±2.14 mg/dL;
p<0.05
), and left ventricular hypertrophy (LVM/ht
2.7
; 31.4 ±6.74
vs
33.5 ±7.15 g/m
2.7
;
p<0.05
), and arterial stiffness (Pulse wave velocity; 4.83 ±0.69
vs
5.35 ±0.92 m/sec;
p<0.05
). Using peripheral blood mononuclear cells, mRNA-Seq, miRNA-Seq, and whole-genome DNA methylation analysis revealed master genes, and regulatory pathways related to BP regulation, tissue fibrosis and cardiovascular remodeling. Our study reveals a novel PH-associated TOD mechanism, showing angiogenesis inhibition mediated by
VASH1
(Vasohibin-1) upregulation and downtrends in
VASH2
(Vasohibin-2),
VEGFC
(Vascular endothelial growth factor C),
HIF1α
(Hypoxia-inducible factor 1-alpha), and
IGF1
(Insulin Like Growth Factor 1). Moreover,
VASH1
targeting miRNA
hsa-miR-30e-5p
is inversely regulated.

Conclusion:
Angiogenesis inhibition in the presence of common demographic and clinical intermediate-phenotypes may contribute to the development of TOD in hypertensive youth.
Abstract 8: Deletion Of The Alpha Subunit Of EnNaC And MTORC2 Inhibition Attenuate DOCA-salt-induced Endothelial Cell And Vascular Stiffness.
Authors: Zhang Liping , Yang Yan , Aroor Annayya , Jia Guanghong , Sun Zhe , Bonnard Benjamin , Ramirez Francisco , Martinez-Lemus Luis A , Jaisser Frederic , Sowers James , Hill Michael A
Published: September 2020

The deoxycortisone acetate (DOCA)/salt model of hypertension and vascular disease replicates a situation of high aldosterone and high salt consumption in humans. We hypothesized that in this model, increased mineralocorticoid receptor (MR) activation would result in increased activity of the endothelial cell sodium channel (EnNaC) leading to increased endothelial cell and vascular stiffness. Female and male mice were implanted with slow release DOCA pellets (50 mg) and given salt in their drinking water (1% NaCl with 0.2% KCl) for 21 days. To probe signaling pathways underlying EnNaC activation induced endothelial cell stiffness we used mice with a specific deletion of the alpha subunit of EnNaC (EnNaC KO) and mice treated with a pharmacological inhibitor of mTORC2 (PP242, 10 mg/kg/day; mTI). Indeed, mTORC2 activation has been shown to increase SGK1 mediated activation of the epithelial Na+ channels in renal epithelial cells (Gleason et al., J. Clin. Invest. 2015). DOCA-salt treated control mice showed an increase in blood pressure and aortic vascular stiffness (ECHO PW analysis) which was attenuated in both the EnNaC KO and mTI-treated groups. Similarly, DOCA-salt administration increased endothelial cell Na
+
transport activity as measured by whole cell patch clamp. The Na
+
conductance was blocked by amiloride consistent with a role for the EnNaC ion channel. Further, intrinsic endothelial cell stiffness as measured by AFM was decreased in EnNaC KO and mTI-treated mice compared to control mice given DOCA-salt, alone. Collectively, the data indicate that in the presence of MR activation and high salt consumption, that activation of EnNaC contributes to endothelial cell and vascular stiffness. This appears to involve a contribution of mTOR signaling which is known to stimulate SGK1 mediated increases in the Na
+
channel at the EC surface and associated increased channel activity. We further conclude that the data are relevant to situations of enhanced aldosterone secretion and MR activation in concert with high salt consumption (ie. salt sensitivitive and resistant hypertension).
Abstract MP04: High Fat Diet Exposure Induces Epigenetic Modulation In The Brain Renin Angiotensin System
Authors: Elgazzaz Mona , Lazartigues Eric
Published: September 2020

Cardiometabolic Diseases (CMD) are one of the highest causes of morbidity and mortality worldwide. In-utero, environmental insults can program brain’s progeny through changes in gene expression level affecting cardiometabolic functions. Methylation and demethylation of DNA take place by DNA methyltransferase (DNMT) and Ten eleven translocation (TET) enzymes, respectively. Expression of renin angiotensin system (RAS) on pre-autonomic neurons in the hypothalamus is believed to play a pivotal role in sympathetic activity controlling metabolic and blood pressure (BP) homeostasis. We hypothesize that perinatal exposure to high fat high sucrose diet (HFHSD) programs the brain RAS through methylation mechanisms that lead to autonomic dysfunction and hypersensitivity to CMD. C57BL6/J dams were fed HFHSD or regular diet (RD) and mated with RD sires. After weaning, offspring were raised on RD. Three-month old progeny underwent 24 h BP recording (telemetry), measurement of fasted blood glucose (FBG), assessment of autonomic function (pharmacological) and baroreflex sensitivity (sequence method). Mice were then sacrificed, and brains harvested for quantification of mRNA levels of DNMT, TET and RAS components in the hypothalamus. HFHSD-exposed male offspring showed elevation in systolic BP (140.2 ±1.9
vs.
131.2 ±2.4 mmHg, p=0.006), heart rate (611.3 ±7.8
vs
562.5 ±7 bpm, p<0.0001) and FBG (172.7 ±6.3
vs.
142.6 ±6.3 mg/dl, p=0.018) but not females. Cardiac sympathetic activity was higher in HFHSD-exposed mice (-148 ±11.5
vs.
-35.2 ±4.7 bpm, p<0.05). Surprisingly, sympathetic vascular tone was only increased in HFHSD-exposed females (-64.6 ±5.7
vs.
-36.7 ±2.9 mmHg, p<0.05) while spontaneous baroreflex gain was improved in HFHSD-exposed males (5.3 ±0.7
vs.
2.6 ±0.4 msec/mmHg, p<0.05). In the brain, DNMT1, DNMT3a and TET1, 2 and 3 mRNA expression were downregulated (p<0.05) while AT
1a
R and ACE2 genes were upregulated (p=0.02) in HFHSD-exposed mice. We conclude that perinatal exposure to a hypercaloric diet epigenetically programs the progeny’s brain through methylation leading to a pre-hypertensive and pre-diabetic phenotype associated with changes in the expression of RAS genes which could eventually result in increased susceptibility to CMD.
Abstract MP05: Pvn-specific Microgliosis And Inflammation Precedes Sympathoexcitation In gαI
2
Protein-dependent, Salt-sensitive Hypertension
Authors: Moreira Jesse D , Chaudhary Parul , Nist Kayla M , Wainford Richard D
Published: September 2020

Aim:
Hypothalamic paraventricular nucleus (PVN) Gαi
2
proteins mediate sympathoinhibitory responses to a high salt (HS; 4% NaCl) diet. Failure to upregulate Gαi
2
proteins in response to a HS diet results in PVN inflammation and salt sensitive hypertension (SSHTN). We hypothesize that microglial-mediated PVN inflammation precedes sympathoexcitation in Gαi
2
protein-dependent SSHTN.

Methods:
Three-month-old male Sprague Dawley rats implanted with ICV cannulas fitted to osmotic minipumps to centrally infuse either a control scrambled (SCR) oligodeoxynucleotide (ODN) or a Gαi
2
targeted ODN, which downregulates CNS Gαi
2
proteins by ~85%, (25μg/5μl/day/ODN) were placed on a 1-7-day normal salt (NS; 0.6% NaCl) or HS diet (n=5/group) and underwent cardiac perfusion. Brain immunohistochemistry was used to assess PVN and subfornical organ microgliosis and qualitatively assess levels of PVN pro-inflammatory cytokines (PIC) IL-1β, IL-6, and TNFα. In additional groups, MAP was assessed via radiotelemetry, and whole blood and kidneys were obtained for ELISA measurement of plasma and renal norepinephrine (NE) as estimates of sympathetic tone.

Results:
By 24h in control SCR ODN infused rats a HS diet, which did not alter MAP or microglial activation, evoked sympathoinhibition. In contrast, in Gαi
2
ODN infused rats a HS diet did not result in sympathoinhibition and evoked significant increases in MAP, PVN microgliosis and PVN PIC expression within 24h, and elevated renal NE content by Day 3.

Conclusions:
Our data suggest that in the male Sprague-Dawley model of PVN Gαi
2
protein-dependent SSHTN PVN inflammation (microgliosis and PIC production) precedes sympathoexcitation.
Abstract MP10: Microbiome-based Diagnostic Screening Of Cardiovascular Disease Using A Machine Learning Approach
Authors: Aryal Sachin , Alimadadi Ahmad , Manandhar Ishan , Joe Bina , Cheng Xi
Published: September 2020

In recent years, the microbiome has been recognized as an important factor associated with cardiovascular disease (CVD), which is the leading cause of human mortality worldwide. Disparities in gut microbial compositions between individuals with and without CVD were reported, whereby, we hypothesized that utilizing such microbiome-based data for training with supervised machine learning (ML) models could be exploited as a new strategy for evaluation of cardiovascular health. To test our hypothesis, we analyzed the metagenomics data extracted from the American Gut Project. Specifically, 16S rRNA reads from stool samples of 478 CVD and 473 non-CVD control samples were analyzed using five supervised ML algorithms: random forest (RF), support vector machine with radial kernel (svmRadial), decision tree (DT), elastic net (ENet) and neural networks (NN). Thirty-nine differential bacterial taxa (LEfSe: LDA > 2) were identified between CVD and non-CVD groups. ML classifications, using these taxonomic features, achieved an AUC (area under the receiver operating characteristic curve) of ~0.58 (RF). However, by choosing the top 500 high-variance features of operational taxonomic units (OTUs) for training ML models, an improved AUC of ~0.65 (RF) was achieved. Further, by limiting the selection to only the top 25 highly contributing OTU features to reduce the dimensionality of feature space, the AUC was further significantly enhanced to ~0.70 (RF). In summary, this study is the first to demonstrate the successful development of a ML model using microbiome-based datasets for a systematic diagnostic screening of CVD.