bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P162: Comparison Of The Clinical Effect Of Empagliflozin On Glycemic And Non-glycemic Parameters In Japanese Patients With Type 2 Diabetes And Cardiovascular Disease Treated With Or Without Baseline Metformin
Authors: Tanaka Atsushi , Node Koichi
Published: August 2021

Introduction:
Recent treatment guidelines for type 2 diabetes (T2D) recommend sodium-glucose cotransporter 2 (SGLT2) inhibitors to be considered preferentially in patients with T2D at high cardiovascular risk or with cardiovascular disease (CVD), regardless of their diabetes status and prior use of metformin.

Hypothesis:
We assessed the hypothesis that the therapeutic impact of SGLT2 inhibitors on clinical parameters differs according to the use of metformin.

Methods:
This was a post hoc analysis of the Effect of Empagliflozin on Endothelial Function in Cardiovascular High Risk Diabetes Mellitus: Multi-Center Placebo-Controlled Double-Blind Randomized (EMBLEM) trial. All participants (n=105; women 31.4%; mean age 64.8 years) had both T2D and DVD and were randomized to either 24 weeks of empagliflozin 10mg daily or placebo. We assessed the effect of empagliflozin on changes from baseline to 24 weeks in glycemic and non-glycemic clinical parameters, according to the baseline use of metformin.

Results:
Overall, 53 (50.5%) patients received baseline metformin. In the 52 patients treated with empagliflozin (48.1% with baseline metformin), the change from baseline systolic blood pressure was greater in patients receiving metformin, compared to metformin-naïve patients (group difference -8.5 [95%CI -17.7 to 0.6 mmHg], p=0.066). Reduction in body mass index was significantly greater in patients receiving baseline metformin, relative to nonusers (-0.54 [95%CI -1.07 to -0.01] kg/m
2
, p=0.047). The group ratio (baseline metformin users vs. nonusers) of proportional changes in the geometric mean of high-sensitivity Troponin-I was 0.74 (95%CI 0.59 to 0.92, p=0.009). No obvious difference was observed in glycemic parameters, such as fasting plasma glucose and HbA1c, between the baseline metformin users and nonusers.

Conclusions:
Our findings suggest empagliflozin has a partially different impact on clinical parameters according to whether or not metformin has been used at baseline. As clinical opportunities for prescribing SGLT2 inhibitors are likely to increase, further research is needed to investigate the effect of SGLT2 inhibitors on clinical parameters taking into account the background situation of hypoglycemic agents, such as metformin.
Abstract P166: Kidney Changes In Patients With Morbid Obesity After Bariatric Surgery: Looking For The Mechanisms
Authors: Oliveras Anna , Vazquez Susana , Galceran Isabel , Goday Alberto , Vera Maria , Soler Maria José , Benaiges David , Crespo Marta , Pascual Julio , Riera Marta
Published: August 2021

Morbid obesity (MO) carries an increased risk of kidney damage. Albuminuria and hyperfiltration decrease after bariatric surgery (BS). The relationships between kidney changes obesity-associated are not fully understood.

Aim:
to analyze renal changes (Δ) and their determinants at 3-mths after BS (3m-postBS) in patients with MO.

Methods:
In a cohort of patients with MO, we analyzed changes in renal function at 3m-postBS and possible associations with anthropometric parameters, ambulatory blood pressure, glucose metabolism, adipocytokine profile, and components of both renin-angiotensin-aldosterone and endocannabinoid systems.

Results:
59 patients were included, 76% women; age (mean ± SD): 42.3 ± 9.5 years; body weight (mean ± SD): 117.8 ± 19.2 Kg. At 3m-postCxB, significant reductions in body weight and waist circumference were observed (p <0.001), but not in blood pressure. Biochemical changes (mean, 95% CI); eGFR-CKDEPI: -4.6 mL/min/1.73m
2
(-8.6; -0.6), p = 0.024; Na
+
: 2.5 mmol/L (1.9; 3.0), p <0.001; K
+
: -0.2 mmol/L (-0.3; -0.1), p = 0.006; HbA1c: -0.47% (-0.63; -0.31), p <0.001 and HOMA-IR-Index: -3.13 (-4.19; -2.06), p <0.001. Δ of Albuminuria: Z-1.8 (p = 0.069). The Δ of eGFR-CKDEPI indirectly correlated only with the Δ of plasma renin activity (PRA), p = 0.026. The Δ of albuminuria indirectly correlated with the Δ of leptin (p = 0.039) and directly with the Δ of HbA1c (0.019), HOMA-index (p = 0.013), ACE2 (p = 0.032) and resistin (p = 0.005), as well as with the Δ of the endocannabinoids N-palmitoyl ethanolamine (p = 0.028) and N-stearoyl ethanolamine (p = 0.022). None of the factors analyzed was associated with changes in sNa
+
. The reduction in sK
+
was significantly correlated with the Δ of leptin (p = 0.028) and with the Δ of aldosterone (p = 0.025). In multivariate analyzes, no factor was independently associated with the different markers of renal function.

Conclusions:
patients with MO experience a decrease in eGFR-CKDEPI associated with an increase in PRA 3m-postBS, indicating that the hyperfiltration present in MO has a hemodynamic origin. On the other hand, the variation in albuminuria is related to the improvement of the carbohydrate metabolism and probably certain cytokines and endocannabinoids have a role, although the latter needs to be confirmed.
Abstract P190: Antihypertensive Treatment Improves Left Ventricular Geometry After 10 Years Even In Elderly Hypertensive Patients
Authors: Toba Ayumi , Ishikawa Joji , Harada Kazumasa
Published: August 2021

Introduction:
Both ageing and high blood pressure (BP) is associated with a risk of left ventricular hypertrophy; therefore, it is unclear whether antihypertensive treatment for 10 years period reduce cardiac hypertrophy and concentricity beyond the impact of ageing. We assessed the hypothesis that even in elderly patients, BP reduction is effective for left ventricular remodeling.

Methods:
Among 558 hypertensive patients without concurrent heart disease referred to a cardiology clinic who underwent echocardiography and ambulatory blood pressure monitoring, 151 patients’ data of echocardiography was available after 10 years of follow up period. We evaluated the relation among BP at baseline, antihypertensive medications, and change in echocardiographic parameters of left ventricular geometry.

Results:
Mean age was 70.8±8.2 years at baseline. 24-hour BP was 127.4±16.4/71.4±9.6mmHg, awake BP was 129.9±17.1/72.4±10.2mmHg, and asleep BP was 122.5±16.9/67.1±9.1 mmHg. After 10 years (mean age 81.0±9.0 years), number of antihypertensive increased from 1.3±1.2 to 2.0±1.1. Left ventricular mass index (LVMI) insignificantly decreased from 115.7 to 98.7mg/m
2
(p=0.167)and relative wall thickness (RWT) significantly decreased from 0.51 to 0.47(p<0.001). There were 12% of patients with normal left ventricular geometry (N), 52% with concentric remodeling (CR), 9% with eccentric hypertrophy (EH), and 27% with concentric hypertrophy (CH) at baseline. Among patients with N or CR at baseline, those who developed to CH or EH had higher night time systolic BP (126.5±17.0 vs 118.9 ±15.4, p=0.037) than those who did not. Neither number nor type of antihypertensive was related to this geometry change. In logistic regression analysis, night time systolic BP was significantly related to the progression of hypertrophy adjusting for age, sex, BMI, LVMI, RWT, and clinic systolic BP at baseline (p=0.04).

Conclusions:
Even in elderly hypertensive patients, antihypertensive treatment for 10 years improved LV geometry in spite of ageing. Nighttime BP at baseline predicted worsening of LV geometry after 10 years beyond clinic BP.
Abstract P191: Association Between Blood Pressure And Brain Volumes Vary By Sex And Age.
Authors: Alateeq Khawlah , Walsh Erin , Abhayaratna Walter , Cherbuin Nicolas
Published: August 2021

Objective:
To quantify the association between blood pressure (BP) across its full range and brain volumes and white matter lesions (WMLs) while investigating the effects of age, sex, body mass index (BMI), antihypertensive medication, and other risk factors.

Methods:
UK Biobank participants (
N
=36,260) aged 40 to 70 years were included and stratified by sex and age into four groups (age ≤ 45, 46-55, 56-65 and > 65 years old). Multi-level regression analyses were used to assess the association between mean arterial (MAP), systolic (SBP), and diastolic (DBP) blood pressure, and brain volumes segmented using the FreeSufer software (gray matter [GMV], white matter [WMV], left [L] and right hippocampal volume [RHCV]) and WMLs. We also investigated the interaction effects between body mass index (BMI) and antihypertensive medication and BP in predicting brain volumes and WMLs.

Results:
Every 10-mmHg higher DBP was associated with lower brain volumes (GMV: -1300– -2800) [SE=34–90]; WMV: -903.44– -1171.7[SE=34.66–53.03]; LHCV: -7.7– -14.26 [SE=0.44–0.57]; RHCV: -6.25– -22.64[SE=0.32–0.95]) across all age groups. A similar pattern was detected in both sexes, although it was weaker, in men. Also, every 10-mmHg higher MAP was associated with larger WMLs across all age groups but peaked > 65 years (0.1 [SE=0.002]). Both lower BMI and anti-hypertensive medication appeared to afford a protective effect.

Conclusion:
Higher BP is associated with worse cerebral health across the full BP range from middle adulthood and across the life course. Therefore, it is important that prevention efforts be directed at younger adults with focus on achieving optimal BP to decrease future risk of developing dementia.
Abstract P194: Voluntary Exercise Eliminates Maternal Gestational Hypertension-induced Hypertensive Response Sensitization (htrs) In Post-weaning High Fat Diet Fed Male Adult Offspring
Authors: Xue Baojian , Yu Yang , beltz terry , Guo Fang , Wei Shun-Guang , Pollock David M , Pollock Jennifer S , Johnson Alan K
Published: August 2021

Exercise has profound effects on cardiovascular function and metabolism in both physiological and pathophysiological states. Our previous studies demonstrated that maternal gestational hypertension (MGHT) induces hypertensive response sensitization (HTRS) elicited by post-weaning high fat diet (HFD) in male offspring. The present study tested whether voluntary exercise would protect against MGHT-induced HTRS in HFD fed male offspring. Male offspring from both normotensive (NT) and MGHT dams were given access to either “blocked” (sedentary offspring) or functional running (exercised offspring) wheels for 10 weeks during normal fat diet (NFD) or HFD feeding. HFD feeding significantly increased resting blood pressure (BP) in sedentary offspring of both NT (112.3±0.7 to 119.9±1.2 mmHg, p<0.05) and MGHT (112.5±0.9 to 129.6±1.0 mmHg, p<0.05) dams, but the elevated BP induced by HFD was greater in sedentary offspring of MGHT dams (129.6±1.0 vs. 119.9±1.2 mmHg, p<0.05). The sedentary offspring of MGHT dams also displayed greater sympathetic tone and enhanced pressor responses to centrally administrated angiotensin (ANG) II or leptin. The running distance was comparable in four groups of exercise offspring (9.183±1.183, 9.192±1.677, 7.233±1.080, 8.482±1.455 kilometers/day, p>0.05). Voluntary exercise did not alter BP in NFD fed offspring and HFD fed offspring of NT dams, but it attenuated BP in HFD fed offspring of MGHT dams (129.6±1.0 to 121.1±0.8 mmHg, p<0.05) and body weight and heart rate in all offspring. Moreover, voluntary exercise significantly reduced sympathetic tone (Hexamethonium, ip, MAP Δ-50.6±1.0 to Δ-29.7±2.7 mmHg, p<0.05) and pressor responses to central ANG II and leptin in HFD fed offspring of both NT (ANG II: Δ16.0±0.9 to Δ7.5±1.1 mmHg; leptin: Δ11.8±0.6 to Δ5.4±0.9 mmHg, p<0.05) and MGHT (ANG II: Δ24.3±2.1 to Δ7.6±1.8 mmHg; leptin: Δ16.8±0.9 to Δ5.2±1.0 mmHg, p<0.05) dams and eliminated the differences in these responses between NFD fed offspring and HFD fed offspring. These results indicate that exercise training plays a beneficial role in preventing MGHT-induced HTRS and that this effect is associated with reduced brain reactivity to pressor stimuli and centrally driven sympathetic activity.
Abstract P198: Biphasic And Sex-dependent Roles Of The Prorenin Receptor In The Rostral Ventrolateral Nucleus In Mice Subjected To Deoxycorticosterone Acetate-salt Hypertension
Authors: Nakagawa Pablo , Brozoski Daniel , Mathieu Natalia M , Lu Ko-Ting , Gomez Javier , Reho John J , Segar Jeffrey , Grobe Justin L , Sigmund Curt D
Published: August 2021

The brain renin angiotensin system (RAS) regulates blood pressure (BP) and autonomic function. However, it remains unclear how and where angiotensin II (Ang II) is generated in conditions eliciting brain RAS overactivation including deoxycorticosterone acetate (DOCA)-salt hypertension (HT). In several tissues, the activation of prorenin requires its binding to the prorenin receptor (PRR). New evidence from this study indicates that prorenin and PRR are co-expressed in the proximity to the rostral ventrolateral nucleus (RVL), an anatomical brain region that controls sympathetic nerve activity. Therefore, we hypothesized that selective ablation of PRR targeting the RVL attenuates BP increase due to DOCA-salt. PRR ablation was targeted to the RVL by stereotactic microinjections of adeno-associated virus (AAV) expressing Cre recombinase-mCherry in PRR-flox mice (PRR
RVL-KO
). AAV mCherry was used as control virus (WT). A pressor response to L-glutamate in the injection site served as confirmatory stereotactic target hit. RVL-targeted ablation of PRR resulted in lower BP responses to DOCA-salt in females (WT=115±3 vs KO=104±4 mmHg;
p
<0.05; n=8), but not males (n=5-8), only during the first 3 days of DOCA-salt treatment. However, at day 13 of DOCA-salt treatment, female PRR
RVL-KO
unexpectedly exhibited exaggerated increase in systolic BP (WT=149±3 vs KO=163±3 mmHg;
p
=0.004; n=8) and pulse pressure (WT=31±4 vs KO=45±4 mmHg;
p
=0.02; n=8) when compared to control. Next, mice were challenged with an intraperitoneal hypertonic saline injection equivalent to 10% of their body weight followed by 4 hours of urine collection. Urinary sodium excretion in female PRR
RVL-KO
was significantly lower when compared to WT (
p
<0.05). These data indicate that the role of PRR in the RVL is sex-dependent and biphasic. That is, PRR contributes to the pressor response during the initial stage of DOCA-salt HT in females, presumably by facilitating the generation of angiotensin peptides in the RVL, while it plays a protective role by promoting renal sodium excretion and preventing elevation of systolic BP during the maintenance stage of DOCA-salt HT. This study suggests that distinct PRR expressing cell populations might elicit diverging physiological functions within the RVL.
Abstract P199: Circadian Contributions To Blood Pressure Variation Under Constant Conditions
Authors: Gloston Gabrielle F , Ensor Annie E , Patel Samarth , Williams Rebecca , Peterson Courtney M , Thomas Stephen J
Published: August 2021

The circadian clock is an endogenous biological timekeeper that responds to environmental changes and governs various physiological processes over a 24-hour cycle. Blood pressure (BP) variation is thought to be controlled by the circadian clock, but few studies have examined circadian control of BP in humans. Moreover, it is unknown whether nighttime BP dipping is driven by the circadian system or by external factors. We investigated whether the circadian system drives 24-hour rhythms in BP, including nighttime BP dipping, using a 30-hour constant routine (CR) protocol. The CR protocol controls for external factors, allowing circadian rhythms to be isolated and measured, by having participants lie in a semi-recumbent posture in dim light (<10 lux) at a constant temperature, consume isocaloric snacks every 2 hours, and maintain wakefulness. To measure the BP rhythm, ambulatory BP was measured every 30 minutes (SpaceLabs 90227), and to measure the central circadian rhythm, core body temperature was measured every 10 seconds using an ingestible, wireless sensor (HQInc Core Body Temperature Wireless Data Record and Sensor). To date, 17 normotensive African American participants (13 females and 4 males), with a mean age of 37 (± 11.3) years and body mass index (BMI) of 32.5 kg/m
2
, have completed the study. Approximately 59% of participants (10 of 17) had non-dipping systolic BP at screening, defined as a <10% decrease in mean systolic BP from daytime to nighttime. Under constant conditions, 94% of participants (16 of 17) had a non-dipping BP phenotype. Median systolic BP dipping was 0.8% for females and 2.2% for males. There was a robust rhythm in participants’ core body temperature but not BP, suggesting that the circadian clock may not contribute substantially to a nighttime decrease in BP in normotensive African Americans. Instead, the non-dipping BP phenotype is likely more so a result of behavioral and/or physiological sleep-related processes. Future research and interventions for non-dipping BP may need to target these underlying behavioral and physiological processes.
Abstract P200: Orthostatic Hypotension Is An Independent Predictor Of Delayed Rehabilitation Recovery
Authors: Okamoto Luis E , Park Jin-Woo , Sharma Purnima , Diedrich Andre , Massey Lauren , Johns Jeffery , Biaggioni Italo
Published: August 2021

Inpatient acute rehabilitation is crucial in the management of patients after injuries, surgery, or those with debilitating or neurological conditions. Orthostatic hypotension (OH) is commonly associated with these conditions and can negatively impact their recovery. We found an incidence of OH of 4% in a cohort of 8350 patients at Vanderbilt Stallworth Rehabilitation Hospital (60±17yr, 72% men vs. non-OH 60±18 yr, 57% men). On admission, OH patients had a longer predicted length of rehabilitation hospitalization stay (LOS; 18±6 vs. non-OH 16±5 days; P<0.01) and lower Functional Independence Measure (FIM, 49±19 vs. non-OH 55±19; P<0.01), reflecting greater medical complexity. Actual LOS was even longer than predicted in OH (actual-predicted LOS [ΔLOS], 1±11 vs. non-OH -2±8 days; P<0.01) and FIM efficiency was lower ([discharge-admission FIM]/LOS, 2.5±1.6 vs. non-OH 3.1±1.9; P<0.01), suggesting that OH may be an independent factor for worse rehabilitation outcomes. To test this hypothesis, we conducted univariate and multivariate regression analyses in the adult patients (n=8146) of this cohort to include other potential factors affecting ΔLOS and FIM efficiency (age, sex, diagnosis of admission and comorbidities grouped into 22 categories by organ systems). We found that OH was independently associated with a longer than predicted LOS (i.e. greater ΔLOS) and lower FIM efficiency (regression coefficient 2.2±0.5 and -0.4±0.1, respectively; P<0.01). These associations remained significant after adjusting for predicted LOS and FIM on admission, suggesting that the effects of OH were not accounted for in these metrics. Other factors significantly associated with greater ΔLOS and lower FIM efficiency were comorbidities associated with the urinary tract, gastrointestinal and peripheral nervous systems, infections, electrolyte imbalance and pressure ulcers. In conclusion, OH has a major independent negative effect on rehabilitation outcomes, and is associated with longer than predicted inpatient rehabilitation LOS and lower functional gain. We suggest that the presence of OH should be considered when setting up the rehabilitation plan, to include management of OH.
Abstract P208: Raas And Markers Of Renal Function In Response To Sleep Hygiene Interventions
Authors: Yang Huan , Vazquez Michael , Eske Annika , Peachthong Emily , Mullington Janet
Published: August 2021

Introduction:
Recent epidemiologic studies report that shorter sleep duration is associated with a rapid decline in renal function. This project investigated the effects of 8 weeks of sleep hygiene interventions on the renin-angiotensin-aldosterone system (RAAS) regulation and markers of renal function. Accumulating evidence suggests that sleep plays an important role in blood pressure (BP) regulation. While BP is influenced by renal function, it is not known if improving sleep hygiene may support regulation of the RAAS.

Methods:
Fifty participants (59.8 ± 1.5 years; 31 women) completed 3 overnight in-hospital stays: baseline (S1), pre-intervention (S2) following a 4-week wait-list evaluation phase from S1 and post-intervention (S3). During S2, participants with elevated BP (BP>120/80 to <160/100) were randomly assigned to two sleep hygiene conditions where in both conditions, the sleep period was stabilized, but in one it was also lengthened. As the study is still ongoing, we remain blind to which condition participants were randomized to. Morning and evening plasma renin activity (PRA), urinary albumin, creatinine, and serum cystatin C levels were measured from all three stays. Albumin-to-creatinine ratio (ACR) and estimated glomerulus filtration rate (eGFR) were calculated.

Results:
There was a significant decrease of night PRA at post-intervention (S3) compared to S2 (p<0.05) and S1 (p<0.05). However, morning PRA did not show any changes throughout the stays (p=0.3). The urinary ACR obtained from evening urine samples did not show significant changes (p=0.66). In addition, serum cystatin C levels measured from evening blood draws and the eGFR did not show significant changes (p=0.21 and p=0.18, respectively).

Conclusion:
The sleep hygiene approach to improving sleep was associated with blunted PRA levels in the evening. When unblinded we will report on whether increasing the sleep period was more effective than stabilizing circadian placement of sleep, alone.
Abstract P211: Single High Na
+
Ingestion Leads To An Increase In Oxidative Stress And Triggers Inflammation.
Authors: Bordcoch Ginette , Tavera Busso Ivan , Masjoan Juncos Juan , Juncos Luis I
Published: August 2021

Hypertension has been linked to a progressive increased in oxidative stress and inflammation. The high prevalence of hypertension poses a great risk to public health as 108 million adults in the United States have the condition. For that reason, a better understanding of the link between a high Na+ intake and the development of hypertension is of crucial importance. We hypothesize that a single ingestion of a high Na+ solution leads to increased oxidative stress and triggers an inflammatory response. Wistar 200-250 g male rats had gastric infusions through the esophagus. Groups were infused with 8 mL liquid Vaseline (Control), 8 mL of NaCl 0.684 M (4% m/v), and 8 mL of NaCl 1.368 M (8% m/v). After infusion, blood was collected at different time points during the first hour. Tissue samples were obtained from the aorta, heart, and kidney. Electron Microscopy (EM) was performed on all tissues, which were also analyzed for molecular markers of oxidative stress: Superoxide Dismutase (SOD) and Malondialdehyde (MDA), and an inflammation marker: Extracellular Signal-Regulated Kinase (ERK). At 2 and a half minutes, serum Na+ concentration was unchanged in the control group compared to an increase observed in animals receiving 4% and 8% Na+ with concentrations of 135±1.4 mEq/L, 141±2.0 mEq/L, and 140±1.2 mEq/L respectively. At the 1-hour time point after infusion, the difference was further increased in the 8% group with serum concentrations of 135±1.8 mEq/L, 140±1.5 mEq/L, and 152±1mEq/L respectively (p<0.05). There was an increase in oxidative stress in the aorta from values of 36.22±4.64 mU/mg SOD and 0.131±0.013 pg/mL MDA in the control group, to 47.11±4.89 mU/mg SOD and 0.291±0.022 pg/mL MDA in the 8% group (p<0.05 in both cases). The same was observed in the heart, where values were: 174.6125.26 mU/mg SOD, 0.026±0.007 pg/mL MDA in controls, and 259.22±21.98 mU/mg SOD, 0.215±0.073 pg/mL MDA in 8% group (p<0.05 both cases). Increased ERK in aortic tissue, values of 0.29±0.03 pg/mL in controls, 2.68±0.18 pg/mL in 4% group and 3.97±0.68pg/mL in 8% group (p<0.05) suggest increased inflammation. We conclude that the elevation in serum Na+ concentration that follows Na+ ingestion leads to increased oxidative stress and inflammation.
Abstract P217: Imbalance Of M1/M2 Macrophages In Ovaries Of Hypertensive Mice Is Associated With Reproductive Dysfunction And Lymphangiogenesis
Authors: Navaneethabalakrishnan Shobana , Wilcox Brooke K , Goodlett Bethany L , Mitchell Brett M
Published: August 2021

Macrophages are the principal immune cells in ovaries. Besides protecting against invading pathogens and antigens, macrophages also play an essential role in folliculogenesis, ovulation, luteinization, lymphangiogenesis, and other functions. An imbalance in M1/M2 macrophages is observed in systemic circulation in patients and animals with hypertension (HTN). Although studies have demonstrated an association between HTN and reproductive dysfunction in females, the effect of HTN on ovarian M1/M2 ratio and lymphatics is largely unknown. We hypothesized that L-NAME-induced HTN (LHTN) and salt-sensitive hypertension (SSHTN) may increase the M1/M2 balance in ovaries which is associated with lymphangiogenesis and reproductive dysfunction in mice. Female mice were either provided L-NAME (0.5 mg/mL) in their drinking water for 3 weeks or L-NAME for 2 weeks, followed by a 2-week washout period and subsequent 3-week 4% high salt diet (SSHTN). Control mice (C) received tap water and normal diet. Flow cytometry analysis revealed a significant decrease in M1 (C: 46%±1, LHTN: 33%±2; p<0.05) and an increase in M2 (C: 7%±1, LHTN: 12%±1; p<0.05) macrophages in ovaries from LHTN mice. In SSHTN mice, ovaries had significantly increased M1 (C: 24%±1, SSHTN: 44%±2; p<0.05) and decreased M2 (C: 12%±1, SSHTN: 4%±1; p<0.05) macrophages. There was a significant increase in gene expression of the hormone receptors AR, FSHR, ERa, ERb, and LHR, and the steroidogenic enzymes StAR, 3bHSD, CYP11a1, and CYP17a1. Ovaries of hypertensive mice had a significant increase in gene expression of the lymphatic vessel markers Lyve-1, Podoplanin, and Prox-1, the lymphangiogenic growth factor VEGF-C and receptors VEGFR-2 and VEGFR-3. Taken together these data demonstrate that HTN disturbs M1/M2 macrophages in ovaries and is associated with reproductive dysfunction and lymphangiogenesis. Manipulation of M1/M2 ratio may be a therapeutic opportunity to improve reproductive health in hypertensive women.
Abstract P219: Losartan Prevents The Recruitment Of Renal M1-like Macrophages And Dendritic Cells In Medulla Of Angiotensin Ii Hypertensive Mice
Authors: Araos Patricio A , Guzmán Andrés , Figueroa Stefanny M , Reyes Javier , Amador Cristián A
Published: August 2021

Immune cells play a major role in the development and progression of hypertension. Previous studies have shown that antigen presenting cells (APCs), such as macrophages (Mø) and dendritic cells (DCs) are particularly abundant in kidney. However, the relevance of these renal APCs on hypertension and whether their distribution change during the anti-hypertensive treatment remain unknow. We evaluated whether losartan (Los) treatment changes the abundance of APCs in the renal cortex and medulla in Angiotensin (Ang) II-infused mice.Male C57BL/6 mice (8-12wo) were treated with AngII (490ng/Kg/min), AngII+Los (20mg/Kg/day) or Vehicle for 14 days (n=4-6). Systolic blood pressure (SBP) was measured by the tail cuff method, and renal cortex/medulla were isolated for the measurements of: APCs (MHC-II
+
:CD11c
+
), DCs (APCs:F4/80
-
:CD64
-
/CD103
+
for type-1 DCs, or APCs:F4/80
-
:CD64
-
:CD11b
+
for type-2 DCs), and M1-like Mø (APCs:F4/80
-
:CD64
+
:CD11b
+
), by flow cytometry.Los treatment prevented the increased SBP (AngII+Los=118.8±6.4 mmHg
vs.
AngII=158.0±21.1 mmHg; p<0.001), and the APCs recruitment in renal cortex (AngII+Los=23.2±2.7
vs.
AngII=36.0±5.9%; p<0.01) and in renal medulla (Veh=16.3±7.7; AngII=26.3±4,7; AngII+Los=14.9±3.3%; p<0.05) induced by AngII. In addition, we observed an increase of DC2 and M1-like Mø recruitments in renal medulla of AngII mice (DC2
Veh
=29.0±5.0
vs.
DC2
AngII
=45.5±7.3%; p<0.05; M1
Veh
=44.8±7.5
vs.
M1
AngII
=58.3±5.3%; p<0.05), which were prevented by Los treatment (DC2
AngII+Los
=27.1±6.8%; p<0.05; M1
AngII+Los
=47.0±3.5%; p<0.05). Interestingly, we did not observe differences between groups on M1-like Mø, and DC2 populations in renal cortex. However, Los treatment prevented the increase of DC1 on renal cortex (Veh=2.1±1.4; AngII=5.2±2.4; AngII+Los=2.1±0.8%; p<0.05), without differences between groups at medullar level.Our results show that Los treatment has a differential effect on the APCs populations in renal cortex and medulla, suggesting that renal APCs have different participations on hypertension according their microenvironment.Supported by Fondecyt #1201251 and #3201016
Abstract P224: Obesity Is Associated With Hypoxia At The Maternal-fetal Interface In A Mouse Model Of Superimposed Preeclampsia, BPH/5
Authors: Adams Daniella , Beckers Kalie , Gomes Viviane , Sones Jennifer L
Published: August 2021

Obesity impacts 1/3 of US adults. Maternal obesity significantly increases risk of adverse pregnancy outcomes, including preeclampsia (PE). Late-gestational hypertension and fetal growth restriction (FGR), hallmarks of PE, are observed spontaneously in BPH/5 mice. Similar to obese preeclamptic women, BPH/5 have increased visceral white adipose tissue (WAT) and are leptin resistant. We hypothesize that attenuation of maternal obesity and reduction of reproductive WAT in pregnant BPH/5 female mice will improve decidualization, placental development, and attenuate FGR. To test this hypothesis, pregnant C57 and BPH/5 female mice fed ad libitum (lib) and pair-fed (PF) BPH/5 female mice, beginning at day of copulatory plug detection, e0.5 (n=5/group), were utilized. Pair-feeding BPH/5 females the equivalent daily food intake of lean control ad lib fed C57 females reduced maternal WAT and circulating leptin in this model by e7.5 as well as reduced pro-inflammatory cytokines, Ptgs2 and IL-6, mRNA in the decidua. Therefore, we tested whether hypoxia-related genes, hypoxia inducible factor (Hif) 1α, heme oxygenase-1 (Ho-1), and stem cell factor (Scf), would be altered in the decidua of PF BPH/5 at e7.5. Using real time PCR, we found that e7.5 implantation sites from ad lib fed BPH/5 had a 1.5 to 2-fold increase in Hif1α, Ho-1, Scf, and VEGF mRNA (p<0.05) compared to C57 that was significantly reduced by PF. Furthermore, real time PCR for leptin (lep) mRNA showed a 6-fold increase in e7.5 implantation sites from ad lib fed BPH/5 versus C57 and lep along with lep receptor mRNA was reduced in PF BPH/5 (p<0.05). Therefore, PF BPH/5 pregnant mice have attenuated obesity and leptin in WAT, circulation, and e7.5 implantation sites that is associated with a reduction in markers of hypoxia at the maternal-fetal interface. Finally, BPH/5 PF litter sizes are significantly higher than ad lib fed, 6 vs 3.5, respectively (n=5-8; p<0.05) and symmetrical FGR is attenuated. In conclusion, maternal weight loss in BPH/5 beginning at conception may improve placental development by mitigating hypoxia at the maternal-fetal interface in this model. Future investigations are needed to determine this effect on the maternal hypertensive syndrome and offspring outcomes long-term.
Abstract P225: Inhibiting B Cell Activating Factor Normalizes The Hypertension And Cytolytic Natural Killer Cells Associated With Placental Ischemia
Authors: Herrock Owen T , Whitney Darby , Amaral Lorena M , Campbell Nathan , Ingram Nicole , Turner Ty , Deer Evangeline , Cockrell Kathy , Fitzgerald Sarah , Lamarca Babbette A
Published: August 2021

Preeclampsia (PE) is new onset hypertension (HTN) during pregnancy and is associated with a pro-inflammatory state with increased T Cells, B Cells, cytolytic Natural Killer (cNK) Cells, and Agonistic Antibodies to the Angiotensin II Type 1 Receptor (AT1-AA) compared to Normal Pregnancy (NP). The Reduced Uterine Perfusion Pressure (RUPP) model of placental ischemia recapitulates these features. We have shown that blocking communication between T Cell CD40L and B Cell CD40 or B Cell depletion prevents HTN and AT1-AA in rat models of placental ischemia induced HTN. This suggests that T Cell dependent B Cell activation is involved in the HTN and AT1-AA seen in PE. B2 B cells maturing into antibody producing plasma cells are the product of T Cell Dependent B Cell Interactions. B Cell activating factor (BAFF) is an integral cytokine in the development of B2 cells specifically. Therapeutic antibodies against BAFF are used in treatment for Systemic Lupus Erythematosus with long-term treatment with Anti-BAFF antibodies reducing patient circulating B cells. Thus, we hypothesize that Anti-BAFF antibodies will selectively deplete B2 cells, therefore reducing HTN and cNK Cells in the RUPP rat model of PE. To test this hypothesis, Gestation Day (GD) 14 pregnant rats underwent the RUPP procedure and some were treated with 1 mg/kg Anti-BAFF antibodies by infusion via jugular catheters inserted 24 hours prior. On GD19, blood pressure (MAP) was measured and circulating, placental, and splenic B Cells and NK cells were measured by flow cytometry. RUPP rats had elevated MAP (119±2 mmHg, n=12) compared to NP (100±3 mmHg, n=9) which was normalized by Anti-BAFF treatment (100±3 mmHg, n=11)(p<0.05). RUPP fetuses were smaller at GD19 (1.97±0.08 g; n=12) compared to NP fetuses (2.207±0.1 g)(p<0.05), and this was unchanged with Anti-BAFF (1.82±0.1 g). RUPP rats had elevated cNK cells (9.1±2%, n=7) compared to NP (3.1±3%, n=3) which was attenuated with anti-BAFF therapy (3.6±1.3%, n=6). RUPP rats had elevated splenic B Cells (4±0.7%, n=8) compared to NP (2±0.7%, n=4) which was reduced by Anti-BAFF (2.8%±1.1%, n=4). In conclusion, Anti-BAFF treatment reduces B cells and cNK cells and normalizes the HTN associated with placental ischemia without worsening fetal weight in a rat model of PE.
Abstract P231: Aspirin Reverses Placental Gene Expression Changes Associated With Preeclampsia
Authors: Santillan Donna A , Devor Eric J , Cooley Brandon J , Pearl Sydney H , Santillan Mark K
Published: August 2021

Aspirin is the only medication recommended for preventing preeclampsia, a hypertensive disorder of pregnancy. However, the mechanism(s) by which aspirin prevents preeclampsia are unknown. To better understand the mechanism by which aspirin prevents preeclampsia, we compared expression of genes involved in signal transduction, cardiac remodeling, ion transport, stress and immune response, apoptosis, sarcomere structure, transcriptional regulation, and cell cycle regulation between placental tissues from 4 different cohorts: preeclamptic women, preeclamptic women who took aspirin during pregnancy, non-preeclamptic women, and non-preeclamptic women who took aspirin (N=3 per cohort). Tissues were obtained from the University of Iowa Maternal Fetal Tissue Bank (IRB# 200910784). When comparing whole thickness samples, 27 genes had a greater than 5-fold change in expression in the tissues of women with preeclampsia compared to those without. For example, CDKN1B whose protein inhibits the cell cycle, is increased in preeclampsia (-3.7ΔΔCt, 13.4-fold increase). However, in women who took aspirin, both those that developed preeclampsia and those who did not, CDKN1B expression was similar to women without preeclampsia who did not take aspirin indicating a decrease in gene expression in response to aspirin. However, aspirin also increases expression of several genes associated with preeclampsia. For example, we identified that RGS2 and PLAC1 in the decidual layer of the placenta were increased in response to aspirin. Decreases in expression of both genes is associated with preeclampsia. However, 19 genes whose expression is increased in preeclampsia compared to controls had no response to aspirin. Identifying specific genes and pathways that serve as targets of aspirin during pregnancy provides valuable insight into the mechanisms by which aspirin reduces the occurrence of preeclampsia and may help to identify who would benefit most from its use. Additionally, it may identify targets for the development of other preeclampsia preventative or therapeutic medications.
Abstract P236: Organofluorine Hydrazones Preventing Oxidant Stress In An
In Vitro
Model Of Preeclampsia
Authors: Zsengeller Zsuzsanna K , Mastyugin Maxim , Lo Agnes , Salahuddin Saira , Karumanchi Ananth , Torok Marianna , Torok Bela
Published: August 2021

Background:
Preeclampsia (PE) is a pregnancy-specific syndrome affecting 5-7% of all pregnant women, and there is no effective treatment available. Early abnormal placental development is associated with oxidative stress and the release of reactive oxygen species in the placenta. This phenomenon leads to downstream signaling and production of anti-angiogenic factors, which cause endothelial and trophoblast dysfunction and the cardinal features of PE, i.e., hypertension, proteinuria, and in severe cases, eclampsia. Our group developed a novel series of mitochondria-targeted antioxidants and sought to test if these compounds can effectively reduce placental oxidative stress and mitigate PE symptoms in vitro.

Methods:
We induced in vitro oxidant stress in human trophoblast (HTR8/SVneo) cells with hydrogen peroxide (H
2
O
2
) and assessed whether augmenting cell-redox function with the proposed antioxidant compounds reduced (i) cell injury (cell cytotoxicity assay), (ii) mitochondrial stress (mitochondrial -derived superoxide production, mitochondria dysfunction) (iii) production of the transcription factor HIF1A and (iv) downstream anti-angiogenic responses (sFLT1 production). These effects were compared with the antioxidants - NAC, Vit E and MitoQ.

Results:
In our cell-based assays, pretreatment with the hydrazone compounds reduced mitochondrial-derived ROS production in H
2
O
2
-exposed trophoblasts cells, indicating that the key factor in the development of PE, the oxidant stress, can be alleviated by the antioxidant hydrazones. The most effective of these compounds, HY12 also reduced HIF-1A expression and sFLT1 protein expression H
2
O
2
-exposed HTR8 cells. Furthermore, the antioxidant hydrazones improved the mitochondrial electron chain enzyme activity in the stressed HTR8 cells, which is another promising characteristic of the applied hydrazones.

Conclusion:
In reducing placental trophoblast oxidative stress, hydrazone-based antioxidants present a potential novel therapeutic approach for the treatment of preeclampsia. Future investigation is warranted regarding the in vivo use of these compounds.
Abstract P239: Identification Of Binding Sites For Phosphodiesterase 5 (PDE5) On Rho-related BTB Domain Containing 1 (RhoBTB1) As A Guide To Identifying Novel RhoBTB1-Binding Partners
Authors: Kumar Gaurav , Lu Ko-Ting , Jing Wu , Fang Shi , Sigmund Curt D
Published: August 2021

In-depth studies of molecular pathways have provided insights to explore therapeutic targets for treating hypertension as a complex disease. We previously identified PDE5 as an interacting partner with RhoBTB1 protein in aortic smooth muscle cells (SMCs). RhoBTB1 regulates PDE5 activity and therefore intracellular cGMP levels, and restoration of RhoBTB1 expression in RhoBTB1-deficient states results in improvement of cardiovascular status in a mouse model of hypertension. Here we identified the specific regions of RhoBTB1 that are responsible for the recruitment of PDE5. Our goal is to use this as a guide to identify additional RhoBTB1 interacting proteins which play important roles in cardiovascular function in the hope their identification may open novel therapeutic avenues against hypertension and associated pathologies. We hypothesize that RhoBTB1 acts as an adaptor for Cullin-3 E3 ring ubiquitin ligase complex, where Cullin-3 functions as “scaffold” which delivers PDE5 (and other targets) to Cullin-3 complex followed by ubiquitination and proteasomal degradation. We generated several truncations in full length RhoBTB1, splitting it into its GTPase, BTB1, BTB2, Carboxyl-terminal (CT), BTB1-BTB2, and BTB1-BTB2-CT domains. To check the preferred binding interface for PDE5, we co-transfected HEK293 cells with epitope tagged PDE5 and separately tagged truncations of RhoBTB1, and analyzed the interaction between PDE5 and RhoBTB1 domains using co-immunoprecipitation assays. Our result shows that BTB1-BTB2-C domain of RhoBTB1 is the preferred binding region for PDE5. The BTB1-BTB2 domain lacking the CT was unable to bind to PDE5. Next, we will utilize stable isotope labeling by amino acids in cell culture (SILAC) followed by proteomic profiling to identify additional targets and/or adaptors involved in the binding of PDE5 and RhoBTB1. Similarly, we will tag specific domains of RhoBTB1 using the ascorbate peroxidase (APEX2) fusion system followed by proteomic profiling.
Abstract P240: Role Of Acetic Acid Produced By Gut Microbiota In The Mechanism Of Reducing Blood Pressure By
Saccharina Japonica
In Renovascular Hypertensive Rats
Authors: Harui Ayaka , Maruyama Saki , Segawa Yukiko , Hashimoto Hiroko , Osera Tomoko , Kurihara Nobutaka
Published: August 2021

In a previous study, we observed that
Saccharina japonica
(SJ) intake reduced blood pressure (BP) in 2-clip, 1-kidney renovascular hypertension model (2K1C) rats. However, the mechanism is not clear. Dietary fibers such as alginate, fucoidan and laminaran are components of SJ and have been reported to change gut microbiota and increase short-chain fatty acids (SCFAs) in normotensive rats. SCFAs have been reported to have an antihypertensive effect when intraperitoneally or intracolonically administered, although it was reported that 2K1C rats reduce SCFAs and bacteria which produce SCFAs compared to sham operated control model (SHAM) rats. To investigate whether acetic acid (AA), one of SCFAs, is involved in the mechanism of BP reduction by an ingestion of SJ, we observed AA, in 2K1C rats fed SJ. Under anesthesia, 2K1C and SHAM were induced to Sprague Dawley rats. Both models were allowed to ingest a standard diet (C group) or a 5% (5 of 100) SJ mixed diet (SJ group) for 7 weeks, to a drinking water freely. During the breeding period, the amount of intake, body weight, and systolic BP (SBP) of each group were observed. After that, rats were euthanized, the contents of cecum were collected, and the pH was measured. The amounts of AA were measured using a commercial kit. The SBP of 2K1C-C was significantly higher than that of SHAM-C throughout the feeding (146 ± 3
vs
131 ± 1 mmHg, p<0.05), the SBP of 2K1C-SJ was significantly lower than that of 2K1C-C (136 ± 5
vs
146 ± 3 mmHg, p<0.05). The pH of the cecal contents was significantly lowered in the SJ group rats in both SHAM and 2K1C. The amounts and concentrations of AA in the cecal contents were significantly increased in both SHAM ( 46 ± 8
vs
92 ± 9 mmol, p<0.05)and 2K1C (50 ± 3
vs
87 ± 5 mmol, p<0.05) in the SJ diet group compared with the C group. On the other hand, there was no significant difference in the amounts of AA between SHAM and 2K1C animal models. These results indicated that ingestion of SJ increased AA, and decreased pH in the cecal contents in both SHAM and 2K1C of the SJ group, which may be involved in the antihypertensive effect of SJ ingestion.
Abstract P243: Inverse Salt Sensitivity In Normotensive Adults: Role Of Demographic Factors
Authors: Romberger Nathan T , Stock Joseph , Patik Jordan , McMillan Ron , Lennon Shannon , Edwards David G , Farquhar William B
Published: August 2021

Inverse salt sensitivity, a sodium induced
reduction
in blood pressure (BP), has recently been associated with increased incidence of hypertension. However, there is a paucity of data regarding the prevalence of inverse salt sensitivity (ISS). Therefore, the purpose of this retrospective analysis was to determine the prevalence of ISS in a cohort of normotensive adults (n=111; age=37±13yr) and to determine if ISS is associated with any demographic characteristic(s). Subjects included healthy, normotensive, and non-obese adults who participated in a controlled feeding study, consuming a low-sodium (LS, 20 mmol Na/d) and high-sodium (HS, 300 mmol Na/d) diet for 7 days each in a randomized order; diet compliance was assessed with 24-hr urinary sodium excretion (LS=29.0±23.6, HS=240.5±92.0, p<0.0001). On the final day of each diet, 24-hr ambulatory BP was assessed. Subjects were grouped based on change in 24-hr mean arterial pressure (MAP) from the LS to HS diet. ISS was defined as a reduction in MAP >5mmHg, salt sensitive (SS) as an increase in MAP >5mmHg, and salt resistant (SR) as a change in MAP ≤5mmHg. Group differences in sex, age, and BMI were evaluated using Chi Square tests and race using a Fisher’s exact test. Group differences in mean age and BMI were assessed using a one-way ANOVA (mean±SD). Overall (n=111), 12.6% (n=14) were ISS, 72.1% (n=80) SR, and 15.3% (n=17) SS. Prevalence of ISS was not different with respect to sex (male [n=57]: 15.8% (9), female [n=54]: 9.3% (5), p=0.30), race (white [n=84]: 14.3% (12), black [n=16]: 6.3% (1), Asian [n=11]: 9.1% (1), p=0.88), or age (21-40 y.o. [n=64]: 15.6% (10), 41-60 y.o. [n=47]: 8.5% (4), p=0.26). However, prevalence of ISS tended to be higher in those with lower BMI (BMI < 25 [n=71]: 16.9% (12), BMI ≥ 25 [n=40]: 5.0% (2), p=0.07). There were also no group differences with respect to mean age (ISS=34±13yr, SR=37±12yr, SS=41±14yr, p=0.29), but SS had a significantly higher BMI than ISS and SR (ISS=22.9±2.8, SR=24.3±2.6, SS=26.7±3.6, p=0.0009). Together, in a normotensive cohort, we found that 12.6% of subjects were ISS. Furthermore, prevalence of ISS was not different with regards to sex, race, and age, but ISS may be associated with lower BMI. Future research is needed to further understand the ISS phenotype.
Abstract P249: Angiotensin At1a Receptor Signaling In Agrp Neurons In Metabolism Control
Authors: Balapattabi Kirthikaa , Mouradian Gary , Hodges Matthew , Sigmund Curt D , Grobe Justin L
Published: August 2021

The adverse cardiovascular responses to leptin are preserved, but beneficial metabolic effects, such asincreases in resting metabolic rate (RMR) are lost in obesity induced by high fat diet (HFD) feeding. Wepreviously reported angiotensin II (ANG) type 1a (
Agtr1a
; AT1A) receptors in a subset of AgRP neurons arecritically required for the integrative control of RMR. Understanding the mechanisms in this unique subset ofRMR controlling- AgRP neurons is the goal of this study and is critical to decipher the pathogenesis of obesity-associated hypertension. Male C57BL/6J mice were maintained on chow or HFD from 8-18 weeks of age. Cell-attached voltage clamp recording of AgRP neurons in ARC sections of Ai9(tdTomato)
AgRP
mice showed that ANGapplication results in three distinct cellular responses (n=65 neurons; 8 mice). In chow fed mice, ANG suppressedelectrical activity in 1/3 of AgRP neurons (ie. “Type 1” AgRP neurons) via a losartan-sensitive mechanism,indicating involvement of AT1 receptors (Firing rate; aCSF:1.23 ± 0.10 vs ANG:0.60 ± 0.08 vs ANG+LOS:1.37 ±0.11 Hz, p<0.05). ANG caused excitation in 1/3 of AgRP neurons (“Type 2” AgRP cells) which was mediatedthrough a PD-123,319-sensitive mechanism, implicating AT2 receptors (ANG:1.69 ± 0.12 vs ANG+PD:0.86 ±0.06 Hz, p<0.05). Finally, 1/3 of AgRP neurons did not respond to ANG (“Type 0” cells). Complementary to thesefindings, mice with AT1A deleted from AgRP cells (ie,
Agrp
-Cre x
Agtr1a
Flox
x Ai9 mice) exhibited a completeloss of Type 1 responses but maintenance of Types 0 and 2 (n=23 cells; 4 mice). Further, pharmacologicaldissection of signaling cascades implicated a pertussis toxin-sensitive mechanism (Gαi cascade; ANG+PTX:1.00± 0.04 Hz, p<0.05) and multiple potassium + chloride channels in the ANG-mediated inhibition of Type 1 cells.Most intriguingly, the relative proportion of AgRP neurons exhibiting Type 1 (ANG -inhibited) vs Type 0 or 2responses was decreased with HFD (Type 1 cells proportion- chow: 35%,23 out of 65; HFD: 18%, 10 out of 56).These results establish a specific ANG-inhibited subtype of AgRP neuron and implicate the AT1A/Gαi signalingcascade in the inhibitory effect of ANG. Switch of Type 1 AgRP cells to Type 0 or 2 in response to HFD suggestsHFD-induced neural plasticity/adaptation.