bluuebunny/crossref_metadata_2025 huggingface dataset:
Association between Gut Microbiota with Mild Cognitive Impairment and Alzheimer’s Disease in a Thai Population
Authors: Wanapaisan Pagakrong , Chuansangeam Mallika , Nopnipa Saifon , Mathuranyanon Rubwad , Nonthabenjawan Nutthawan , Ngamsombat Chanon , Thientunyakit Tanyaluck , Muangpaisan Weerasak
Published: September 2022

Background: Mild cognitive impairment (MCI) and Alzheimer’s disease (AD) are common in older adults. Much recent work has implicated the connection between the gut and the brain via bidirectional communication of the gastrointestinal tract and the central nervous system through biochemical signaling. Altered gut microbiota composition has shown controversial results based on geographic location, age, diet, physical activity, psychological status, underlying diseases, medication, and drug use. Objectives: This study aimed to investigate the relationships of gut microbiota with MCI and AD. Methods: 16S metagenome profiles from stool collection of participant groups (normal; n = 20, MCI; n = 12, AD; n = 20) were analyzed. The diagnosis of cognitive conditions was made by standard criteria consisting of clinical interviews, physical examinations, cognitive assessments, laboratory examinations, and neuroimaging by both structural neuroimaging and amyloid positron emission tomography scans. Correlations between medical factors with food frequency and the fecal microbiome were elucidated. Results: A significant difference at the operational taxonomic unit level was observed. The significantly higher abundance of bacteria in nondementia patients belonged to the Clostridiales order, including Clostridium sensu stricto 1 (p < 0.0001), Fusicatenibacter (p = 0.0007), Lachnospiraceae (p = 0.001), Agathobacter (p = 0.021), and Fecalibacterium (p < 0.0001). In contrast, Escherichia-Shigella (p = 0.0002), Bacteroides (p = 0.0014), Holdemanella (p < 0.0001), Romboutsia (p = 0.001), and Megamonas (p = 0.047) were the dominant genera in the AD group. Left and right hippocampus and right amygdala volumes were significantly decreased in the AD group (p < 0.001) and significantly correlated with the groups of bacteria that were significantly different between groups. Conclusion: There was a relationship between the composition of the gut microbiome and neurodegenerative disorders, including MCI and AD. Reduction of Clostridiaceae and increases in Enterobacteriaceae and Bacteroides were associated with persons with MCI and AD, consistent with previous studies. The altered gut microbiome could be potentially targeted for the early diagnosis of dementia and the reduction of AD risk.
Abstract 016: Magnesium Deficient Diet-induced Hypertension And Renal DC-specific NLRP3 Inflammasome Activation
Authors: Pitzer Ashley L , Kirabo Annet , Ray Evan C
Published: September 2022

Background:
Analysis from a 2013-2016 dietary survey by the National Health and Nutrition Examination Survey Several found that less than 50% of Americans consume sufficient magnesium. Multiple studies have identified a link between dietary magnesium and inflammation associated with multiple pathologies including hypertension, chronic kidney disease, and cardiovascular disease. Additionally, magnesium sulfate inhibits NLRP3 inflammasome activation in human macrophages. We have previously shown that activation of the NLRP3 inflammasome is a key component of salt-sensitive hypertension. In this study, we hypothesize that a magnesium-deficient diet could promote NLRP3 inflammasome activation in myeloid cells and systemic hypertension.

Materials and Methods:
In vivo
studies were performed on mice with an
SV/129
background given either a normal-magnesium (Norm-Mg
2+
) diet (0.08%, as magnesium oxide) or low-magnesium (Low-Mg
2+
) diet (0.01%) for 5 weeks. Systolic blood pressure (SBP) was monitored on a weekly basis by tail-cuff.

Results:
Mice fed a Low-Mg
2+
diet had a significant increase in SBP compared to the Norm-Mg
2+
diet fed mice (105.8 ± 11.3 to 130.4 ± 7.4 mmHg vs 106.5 ± 9.4 to 121.8 ± 6.5 mmHg, [n=7, p=0.0071, 2 way ANOVA]). Using flow cytometry, we observed an increase in NLRP3 (34.1 ± 2.5 vs 7.4 ± 0.6 % of DCs, [n=5-7, p=0.0025, Mann-Whitney test]) and IL-1β (41.9 ± 6.6 vs 15.86 ± 6.6 % of DCs, [n=5-7, p=0.048, Mann-Whitney test]) expression in renal dendritic cells (DCs) of Low-Mg
2+
mice compared to Norm-Mg
2+
mice. There was a similar increase in IsoLG-adduct formation in renal DCs (8.5 ± 2.0 vs 0.6 ± 0.05 % of DCs, [n=5-7, p=0.0025, Mann-Whitney test). Interestingly, we found no difference in NLRP3 (24.92 ± 5.3 vs 31.54 ± 3.1 % of monocytes, [n=5-7, p=0.1376, Mann-Whitney test]), IL-1β (9.5 ± 4.3 vs 4.6 ± 0.4 % of monocytes, [n=5-7, p=0.7551, Mann-Whitney test]), or IsoLG-adduct formation (29.42 ± 3.6 vs 31.9 ± 3.6 % of monocytes, [n=5-7, p=<0.999, Mann-Whitney test]) in renal monocytes.

Conclusions:
Our findings suggest a magnesium-deficient diet induces a hypertensive response and activates the NLRP3 inflammasome in renal DCs but not monocytes.
Abstract 019: T Cell Recognition Of Isolevuglandin-adducted Proteins In Hypertension Is Mhc Class 1-restricted
Authors: Bloodworth Nathaniel C , Chen Wei , Ruggeri Barbaro Natalia R , Ao Mingfang , Palubinsky Amy M , O'Niel Richard T , Phillips Elizabeth J , Moretti Rocco , Davies Sean S , Mallal Simon A , Meiler Jens , Harrison David G
Published: September 2022

Introduction:
Isolevuglandins (isoLGs) are peroxidized lipids that covalently bond lysine residues to modify self-antigens in hypertension. We hypothesized that isoLG-adduction restricts peptide presentation to specific class 1 major histocompatibility complexes (MHC) in mice and humans.

Methods and Results:
We developed 2 mouse strains expressing either his-tagged class 1 MHC-I H-2D
b
or H-2K
b
with truncated membrane binding domains. Shed MHC-I from cultured splenoctyes was adsorbed onto nickel-agarose beads and used to stimulate T cells. We found that CD8+ T cell proliferation is restricted to H-2D
b
and not H-2K
b
(Fig 1A) and occurs only if both T-cells and soluble H-2D
b
are from hypertensive (angiotensin II-treated) mice. Proliferation was blocked if donors received an isoLG-scavenger or by an isoLG specific antibody added during the proliferation assay (Fig 1B). Fluorescence resonance energy transfer (FRET) demonstrated that isoLG-adducts associate with MHC-1 H-2D
b
and not H-2K
b
. We transfected HLA-null K562 cells with 18 common human HLA subtypes, and induced isoLG formation with tert-butyl hydroperoxide. Using FRET we identified human HLAs that are high, intermediate and low presenters of isoLG adducts (Fig 1C). Unique modeling software identified candidate peptides and showed that structural characteristics of H-2D
b
cause lysines at positions 4-7 to project from the MHC groove and present isoLGs (Fig 1D-E).

Conclusion:
IsoLG adduct presentation is MHC-1 restricted. This may explain differences in the degree of inflammation and end-organ damage observed between populations with hypertension and provides a possible new approach to personalized care.
Abstract 030: Aldosterone Promotes Nanoscale Colocalization Of Epithelial Sodium Channel Subunit Alpha And Sodium Calcium Exchanger In Neuro2a Cells
Authors: Aliabadi Simindokht , Akin Elizabeth , Krishnan Vivek , Earley Scott , Feng Earley Yumei
Published: September 2022

We previously reported that hypertensive patients have a higher concentration of sodium ([Na
+
]) in their cerebrospinal fluid (CSF) compared with normotensive subjects but no difference in plasma [Na
+
] was observed, suggesting the potential importance of CSF [Na
+
] in hypertension. Epithelial sodium channels (ENaC) in the brain have been shown to be up regulated in salt-induced hypertension (SH). However, the molecular mechanisms of how brain ENaC contributes to SH development are not yet understood. We hypothesize that ENaC forms a nanoscale colocalization with the sodium-calcium exchanger (NCX) that mediates downstream signaling pathways leading to SH. To test our hypothesis, using self-labeling proteins we generated SNAP-ENaC-α and Halo-NCX1 constructs, enabling high-resolution imaging of surface expressed proteins. Expression of ENaC-α-SNAP and NCX1.5-Halo was confirmed by western blot showing an increase in expression (fold change) of ENaC-α (7.6 ± 1.7 vs. 1 ± 0.3, p<0.02) and NCX1.5 (10.1 ± 0.8 vs. 1 ± 0.4, p<0.0006) in transfected Neuro2A cells compared with controls. Transfected cells were treated with vehicle or aldosterone (100 nM) for 2 hours (n=15 cells from 3 repeat experiments/group) and imaged using ground-state depletion super-resolution microscopy. Data were analyzed using object-based analysis (OBA). Aldosterone treatment increased the density of individual ENaC-α clusters on the cell surface (12.4 ± 1.3 vs. 6.9 ± 1.08 clusters/μm
2
, p<0.002), supporting the functional role of aldosterone on increasing ENaC-α surface abundance. Importantly, the frequency (3.1 ± 0.34% vs 1.7 ± 0.22% colocalized /total clusters, p<0.001) and the density (0.5 ± 0.1 vs 0.2 ± 0.04 clusters/μm
2
of cell surface, p<0.009) of colocalized (< 40nm) ENaC-α and NCX 1.5 clusters were significantly increased in aldosterone treated cells. In sum, these data show aldosterone increases the number of ENaC-α channels on the cell surface and promotes its nanoscale colocalization with NCX, providing evidence for a functional role of ENaC-α and NCX1 coupling. Nanoscale colocalization of ENaC and NCX is potentially significant in SH, and future studies will focus on elucidating the functional importance and mechanisms.
Abstract 034: Absence Of Tachyphylaxis And Vasorelaxant Effect Induced By Nono2p, A New Nitric Oxide Donor
Authors: dos Anjos Moraes Raiana , Alexandra de Araujo Fênix , Barreto da Silva Liliane , Leonne Cruz de Jesus Rafael , Santana de Brito Daniele , Santos de Sá Denise , Daniel Silva da Silva Carlos , Novais da Rocha Zenis , Priviero Fernanda , Webb R C , Flavia Silva Darizy
Published: September 2022

Introduction:
Decreased bioavailability of nitric oxide (NO) plays a mechanistic role in hypertension and myocardial infarction. NO donors are potent vasodilators, but often exhibit toxicity or vascular tolerance. The aim of this study was to investigate the vascular activity possible tachyphylaxis of NONO2P, a novel NO donor.

Methods:
Male Wistar rats were euthanized, and the superior mesenteric artery was isolated for recordings of isometric force, and
in vivo
experiments were performed to evaluate blood pressure in non-anesthetized normotensive rats.

Results:
Cumulative administration of the NONO2P (10
-13
to 3x10
-6
M) induced endothelium-independent relaxation (Emax:111.51 ± 2.31%; pD2: 8.51 ± 0.08, n=9) in arterial rings pre-contracted with phenylephrine (Phe,1μM). However, the relaxation was reduced in pre-contracted rings with Phe exposed to Tyrode's solution containing 20 mM of K
+
(E
max
: 102.83 ± 2.10%; pD2: 7.73 ± 0.04, n=6), suggesting the participation of K
+
channels in the relaxation. The presence of the specific soluble guanylyl cyclase (sGC) inhibitor, ODQ (10μM), abolished the vasorelaxant effect (E
max
: 15.38 ± 11.85%, n=7). Pre-incubation with cyclopiazonic acid (CPA) (10μM), inhibitor of sarcoendoplasmic reticulum calcium ATPase (SERCA), shifted the relaxation concentration-response to the right (E
max
: 106.17 ± 3.06%; pD2: 7.69 ± 0.04, n=6). Interestingly, repeated NONO2P administration did not induce tachyphylaxis, and NONO2P presented similar maximum efficacy to sodium nitroprusside (SNP) (E
max
: 114.24 ± 3.47%; pD2: 9.40 ± 0.04, n=6). Moreover, NONO2P lowered blood pressure in normotensive rats.

Conclusion:
The endothelium-independent vasorelaxant effect induced by NONO2P involves both sGC, SERCA and K
+
channel activation, and NONO2P does not appear to cause tachyphylaxis. NONO2P is able to promote vasorelaxation with the same magnitude as SNP. Finally, NONO2P reduces blood pressure, becoming a promising molecule as a novel therapeutic alternative for the treatment of cardiovascular diseases.
Abstract 036: Failure To Achieve Optimal Blood Pressure Control Among Stroke Patients In The Outpatient Setting
Authors: Sreekrishnan Anirudh , Lansberg Maarten G , Low Yen , Manickam Raj , Kraler Lironn
Published: September 2022

Introduction:
Despite being an AHA Class 1 recommendation to achieve a blood pressure (BP) < 130/80 for secondary stroke prevention, only an estimated 20% of the general population can achieve this benchmark. Furthermore, it has been suggested that stroke patients may be at higher risk for an inability to achieve this goal. We aimed to examine real world clinic data to evaluate BP control among stroke patients.

Methods:
We conducted a population-based retrospective study using electronic health records collected during routine care at our institution for all adult patients 18 years or older with a diagnosis for stroke between January 2019 to November 2021. BP measurements were taken in an outpatient setting between 90 and 180 days from the index stroke. Descriptive statistics using R (R Core Team, 2022, r-project.org) were reported as the mean and standard deviation for continuous variables and frequencies and proportions for categorical variables.

Results:
Our review identified a total of 1,583 patients with a new diagnosis of acute ischemic stroke (AIS: 1,252; 79.1%) or intracerebral hemorrhage (ICH: 331; 20.9%). AIS patients were 45.7% female with a mean age of 68 years old, compared to 47.7% female and 59 years old for ICH patients. AIS patients had a higher proportion of secondary medical issues, including 34.7% with diabetes, 33.0% congestive heart failure, 19.2% cardiac disease, and 33.9% renal disease (compared to 22.2%, 16.1%, 8.8%, 19.2% respectively for ICH patients). In follow-up, only 11.7% of AIS patients were at goal (defined as a BP < 130/80) at 3 months, 17.3% at 6 months, 11.6% at 9 months, and 8.9% at 12 month follow-up. Conversely, ICH patients were slightly better controlled with 21.6%, 32.8%, 15.8%, and 12.2% controlled at 3, 6, 9, and 12 month follow-up respectively. Patients with stroke after March 2020 (post-COVID19 pandemic) had lower rates of blood pressure control compared to those diagnosed one year prior to the pandemic.

Conclusion:
Patients with AIS have lower rates of BP control compared to the general population, which was further amplified by the COVID19 pandemic. These results may suggest that stroke patients face unique barriers in BP management and highlight the need to perform targeted treatment for this especially vulnerable group.
Abstract 042: A Novel (pro)renin Receptor Variant Prr24 As A Specific Renin Receptor And Key Regulator Of Nkcc2 And Urine Concentrating Capability
Authors: Yang Tianxin , Chen Yanting , Feng Ye , Wang Fei , Zheng Huaqing , Fu Ziwei , Zou Chang-Jiang
Published: September 2022

(Pro)renin receptor (PRR) has emerged as a key regulator of renal handling of Na
+
and water, and blood pressure (BP). Renal action of PRR is at least in part mediated through its ability to activate tissue prorenin and renin. In preliminary studies, investigation of a “non-specific” protein band recognized by an anti-PRR antibody against the extracellular domain led to identification of a novel 24-kDa isoform of PRR, termed as PRR24. PRR24 was encoded by a novel cDNA with part of 5’UTR arising from retention of intron 3 and an ATG in exon 5 as an alternative start codon so that this isoform overlaps completely with PRR except that N-terminal 136 amino acids are missing. PRR24 selectively bound and activated renin but not prorenin whereas PRR non-selectively interacted both prorenin and renin. PRR24 protein and mRNA expression was exclusively detected in the renal medulla but not renal cortex or extrarenal organs. Within the kidney, PRR24 mRNA was restricted to medullary thick ascending limb (mTAL) as assessed by RT-PCR on microdissected nephron segments. Using CRISP/Cas9 strategy, we generated a mouse model with mutagenesis of the start codon in exon 5 (termed PRR24
mut
) by replacing methionine with glutamine in PRR. Renal medullary protein expression of PRR24 was barely detectable contrasting unchanged PRR abundance in the null mice. PRR24
mut
mice was born at Mendelian ratio and didn’t exhibit developmental abnormalities or mortality as seen in systemic PRR KO mice. PRR24
mut
mice developed polyuria, hypoosmotic urine, polydipsia, hypotension (radiotelemetry MAP: WT 120
+
0.8 vs. Mutant 112 + 1.4 mmHg; p<0.05). Plasma volume assessed indirectly by hematocrit and directly assessed by using a fluorescent tracer was consistently reduced by 5-10%. Renal expression of NKCC2 was downregulated by 70% in parallel with reduced diuretic response to furosemide. Following 24-h water deprivation, the upregulation of NKCC2 expression was blunted in PRR24
mut
mice associated with impaired urine concentrating capability as assessed by urine volume and osmolality. Overall, PRR24 is a novel PRR variant and serves as a specific renin receptor to regulate renal NKCC2 expression to ultimately determine urine concentrating capability.
Abstract 044: Stimulation Of Intrarenal Angiotensinogen Expression And The Development Of Hypertension And Kidney Injury In Angiotensin II-infused Hepatic Angiotensinogen Knockout Mice
Authors: Satou Ryousuke , Ye Dien , Lu Hong S , Katsurada Akemi , Dugas Courtney M , Daugherty Alan , Motazedian Shahriar , Navar L G
Published: September 2022

Regulation of systemic and local angiotensinogen (AGT) levels is a key determinant of tissue angiotensin II (Ang II) levels and inappropriate AGT augmentation promotes the development of hypertension and tissue injury. Kidney and urinary AGT levels are increased in Ang II-mediated hypertension. Recent studies have demonstrated that circulating hepatocyte-derived AGT (hAGT) enters kidneys sustaining kidney and urinary AGT levels. However, roles of hAGT in blood pressure elevation and kidney injury in Ang II-mediated hypertension have not been delineated. This study tested if hAGT contributes to the development of the pathophysiological events in Ang II-infused mice. A low dose of Ang II (400 ng/kg/min) was infused to male wild type (WT) and hAGT gene knockout (KO) mice (N=9 and 13) for 4 weeks. The control group in each genotype received vehicle (Veh) infusion (N=5 and 6). Western blot confirmed non-detectable levels of hAGT in KO mice. hAGT KO markedly decreased plasma AGT levels (WT+Veh:12.2±0.6 vs. hAGT KO+Veh: 0.8±0.1 μg/ml). Ang II infusion did not elevate plasma AGT levels in either WT and hAGT KO mice. Although hAGT KO mice exhibited a lower baseline of systolic blood pressure (SBP) than WT mice, Ang II-mediated increases in SBP was not attenuated in hAGT KO mice (ΔSBP in WT+Ang II: 30.1±4.4 vs. hAGT KO+Ang II: 26.0±4.2 mmHg). Kidney AGT mRNA levels were increased by Ang II infusion to the same extent in both WT and hAGT mice (WT+Ang II: 1.30±0.04 vs. hAGT KO+Ang II: 1.34±0.06, ratio to control). Likewise, Ang II infusion increased IL-6 mRNA to the same magnitude in both WT and hAGT KO mice. Urinary AGT was sustained in hAGT KO+Veh mice (66±9%) compared to WT+Veh mice. Ang II infusion did not alter urinary AGT levels in both groups. Glomerular mesangial expansion and fibrosis by Ang II infusion were not observed. Ang II infusion developed tubulointerstitial fibrosis in renal cortex and medulla. hAGT KO prevented the fibrosis only in the medulla. These outcomes demonstrate that elevation of SBP, augmentation of intrarenal AGT and IL-6 expression, and the development of renal cortical fibrosis in Ang II-mediated hypertension do not require hAGT. In contrast, hAGT contributes to renal medullary fibrosis which may be due to the lower absolute levels of blood pressure.
Abstract 058: ACE2 Ubiquitination Regulates Neuronal Activity In Hypertension
Authors: Mohammed Mazher , Elgazzaz Mona , Yue Xinping , Filipeanu Catalin , Lazartigues Eric D
Published: September 2022

ACE2 expression on GABAergic neurons could maintain the inhibitory tone over pre-sympathetic neurons in the paraventricular nucleus (PVN) and thus contribute to the regulation of blood pressure (BP). Here we investigated the role of ACE2 ubiquitination in the bed nucleus of the stria terminalis (BNST), a region containing GABAergic neurons and involved in cardiovascular responses to stress. C57BL6/J male mice were injected in the BNST with a ubiquitination-resistant human ACE2 mutant (AAV-hACE2-5R). A subset of mice was also injected with AAV-GCaMP6s-eYFP in the PVN and BNST to record intracellular Ca
2+
activity via fiber photometry. After 6 weeks, the mice were chronically infused with Ang-II (600 ng/kg/min) for 2 weeks while BP was monitored by telemetry. Ang-II-infusion resulted in a significant increase in mean BP compared to controls (152±7
vs.
123±4 mmHg, respectively; n=6, p<0.05) that was blunted in mice expressing hACE2-5R (133±5 mmHg, n=6, p<0.05). Injection of bicuculline (GABA
A
antagonist, 1 mmol/L in 100nL) in the PVN produced a blunted BP rise in Ang-II-infused mice compared to controls (AUC: 261±36
vs.
392±45, respectively; n=4, p<0.05) and an enhanced pressor response in the hACE2-5R+Ang-II group (AUC: 690±81, n=5, p<0.05). Acute injection of Ang-II (250 ng iv) caused an immediate increase in Ca
2+
mediated fluorescence in the PVN and a parallel reduction of neuronal activity in the BNST (ΔF/F
0
: +0.9±0.02 and -0.1±0.03; respectively). Surprisingly, hACE2-5R expression in the BNST produced a rapid and persistent inhibition of neuronal activity. Capillary Western analysis showed a 3-fold increase in NEDD4-2 (n=4, p<0.05) with a parallel reduction of ACE2 levels (-50%, n=4, p<0.05) in the BNST of Ang-II-infused mice, while hACE2-5R prevented NEDD4-2 upregulation and ACE2 downregulation (n=4, p<0.05). Our data show that increases in peripheral Ang-II levels result in an immediate regulation of neuronal activity in the PVN and BNST. Hypertension is associated with NEDD4-2 mediated ubiquitination of ACE2 blunting GABAergic input to the PVN and potentially increasing the activity of pre-sympathetic neurons. ACE2 ubiquitination by NEDD4-2 in key brain regions is a new mechanism contributing to the development of neurogenic hypertension.
Abstract 065: Decreasing Blood Pressure In Mice With Salt-Sensitive Hypertension Changes Renal Immune Cell Populations
Authors: Smith Hannah , Navaneethabalakrishnan Shobana , Goodlett Bethany , Mitchell Brett M
Published: September 2022

Salt-induced hypertension is associated with chronic interstitial inflammation and increases in activated inflammatory immune cells in the kidney. It is currently unknown if these renal immune cell changes result from high blood pressure or directly from high salt concentrations. Using a salt-sensitive hypertension (SSH) mouse model and hydralazine (HDZ), we hypothesized that some renal immune cells are affected by high blood pressure, and some will be affected by the high salt content without increased systemic pressure. Three groups of mice (n=5 in each) were generated and given L-NAME via drinking water for 2 weeks, with two groups receiving a high salt (4%) diet for 3 weeks after a 2-week washout, and one of these receiving HDZ (250 mg/L) via drinking water simultaneously during the high salt diet. Systolic blood pressure was significantly decreased in the SSH group treated with HDZ, dropping to 119±1 mmHg from 141±1 mmHg (p<0.001) in the untreated SSH group. Kidneys were immunophenotyped via flow cytometry, and had decreased M2 macrophages (SSH: -55%, p=0.012; SSH+HDZ: -58%, p=0.0096), increased Th17 cells (SSH: 60%, p=0.0015; SSH+HDZ: 47%, p=0.0086), and increased natural killer cells (SSH: 223%, p=0.013; SSH+HDZ: 276%, p=0.0031). In the SSH group only, Th1 cells increased 50% (p=0.047), Th2 cells increased 67% (p=0.035), dendritic cells increased 31% (p=0.025), and CD3+ T cells decreased -46% (p=0.0092). qPCR of collected kidney tissue revealed a three-fold increase in the macrophage/lymphatic marker Lyve1 (p=0.014) only in the SSH group. These results imply that SSH plays a significant role in shaping renal immune cell populations and renal inflammation. Specific renal immune cell subsets undergo changes due to both the high salt and high systemic pressure while others undergo changes only due to the high salt. Future studies will evaluate the therapeutic potential for targeting these immune cell populations and the effect it has on salt-induced hypertension.
Abstract 076: Sex And Stress In Fructose-induced Salt-sensitive Hypertension
Authors: Brostek Autumn , Hong Nancy J , Zhang Ronghao , Kaydo Lindsey , Kluge Nicholas , Smith Corey , Garvin Jeffrey , Gonzalez-Vicente Agustin
Published: September 2022

Proximal tubule fructose metabolism is key to fructose-induced hypertension, but the roles of sex and stress are unclear. We hypothesized that females are resistant to the salt-sensitive hypertension caused by low amounts of dietary fructose compared to males and that the magnitude of the increase in blood pressure (BP) depends, in part, on amplification of the stress response of renal sympathetic nerves. We measured systolic BP (SBP) in rats fed high salt with either no sugar (HS), 20% glucose (GHS) or 20% fructose (FHS) in the drinking water for 7-8 days. FHS increased SBP (p<0.03 vs basal) but neither GHS nor high salt alone raised SBP. FHS increased SBP significantly and similarly in both (male: Δ25±8 mm Hg; female: Δ19±2 mm Hg). FHS increased SBP by 24±5 mm Hg but only by 8±2 mm Hg when measured by plethysmography and telemetry, respectively (p<0.004). When SBP was measured by telemetry under low stress, FHS increased SBP by 8±1 mm Hg; on the contrary, when measured by telemetry under moderate stress conditions (simulating stress of plethysmography), FHS increased SBP by 15±3 mm Hg, a significantly greater increase (p<0.008). Norepinephrine excretion in rats subjected to moderate stress was 63±17 nmole/Kg/day for animals fed FHS but only 19±40 nmole/Kg/day for controls fed HS (p<0.02). We conclude that fructose-induced salt-sensitive hypertension is similar in males and females unlike other forms of hypertension, and the increase in blood pressure depends in part on an augmented response of the sympathetic nervous system to stress.
Abstract 077: 17b-estradiol Implants With An Antiandrogen Or Castration Attenuate Body Weight And Total Lean Mass But Differentially Modulate Total Cholesterol And The Pressor Response To Chronic Angiotensin II In The Male Rat
Authors: Alexander Barbara T , Rawls Adam Z , Cockrell Kathy , Davenport Kacey , Crudup Breland F , McNeal Miranda J , Rafique Jasmine F , Ojeda Norma B , Shawky Noha M
Published: September 2022

Background:
Transfemales are biological males at birth, but identify as female by gender. Gender-Affirming Hormone Therapy (
GAHT
) is used to alter hormone levels to match gender identity. GAHT in transfemales involves administration of estradiol (
E2
) with an androgen antagonist (
AA
) or orchiectomy. Sex is an independent predictor of cardiovascular (
CV
) risk, but the role of sex hormones on CV risk in transfemales is unknown. Thus, this study tested the hypothesis that GAHT in male rats increases CV risk.

Methods:
Male Sprague Dawley (
SD
) rats were randomly assigned at 14 weeks of age to three groups that underwent either mean arterial pressure (
MAP
, N=3-5/group) or metabolic (N=8/group) monitoring. Control (
C
): empty silastic capsule.
E2+AA:
silastic capsule with 17-beta E2 benzoate (5 mg/21 days) plus AA, spironolactone (10 mg/kg/day, incorporated in diet).
E2+CTX:
silastic capsule with E2 plus castration (
CTX
). Silastic capsules (+/- E2) were replaced every 21 days starting at 14 weeks. AA, started at 14 weeks of age, was continued in E2+AA but discontinued after CTX at 16 weeks of age in E2+CTX.

Results:
(Table 1)
:
The pressor response to angiotensin II (
ANG II
) was similar in C and E2+CTX but was blunted in E2+AA despite similar baseline MAP, increase in E2 and decrease in testosterone. Cholesterol was only increased in E2+AA.

Conclusion:
These results suggest that feminizing therapy in male rats is associated with similar changes in sex steroids but the pressor response to ANG II and lipid profile differ based on the method of androgen reduction. The use of a transfemale rodent model will allow study of the pathophysiology of increased CV risk associated with GAHT across the lifespan.
Abstract 078: Human Gain-of-function Mutation W521r In The Enac α-subunit Contributes To Salt-sensitive Hypertension In A Sex-specific Manner
Authors: Ishimwe Jeanne , Saleem Mohammad , Kleyman Thomas R , Ray Evan C , Kirabo Annet
Published: September 2022

The epithelial Na
+
channel (ENaC) plays a key role in regulating the extracellular fluid volume and blood pressure. Gain-of-function mutations in ENaC subunits are associated with hypertensive disorders including Liddle syndrome. We previously demonstrated that in male mice, dendritic cell ENaC mediates NADPH oxidase activation and increased accumulation of Isolevuglandins (IsoLGs), which act as neoantigens, promoting T cell activation and salt-induced hypertension. The mechanism by which dendritic cell ENaC is regulated, and the role of sex as a biological variable is still yet to be defined. One of the mechanisms by which ENaC is regulated is through sodium self-inhibition. We hypothesized that sodium self-inhibition plays a sex-specific role in regulating ENaC leading to salt-sensitive hypertension. To test this hypothesis, we generated mice with a gain-of-function mutation of a single nucleotide variant (W521R) in ENaC α-subunit using CRISPR/Cas9 in the wild-type (WT) salt-sensitive129/sv background. These mutant mice lack the sodium self-inhibition of ENaC. WT littermate male and female mice were used as controls. Using telemetry blood pressure monitoring, we found that WT males developed salt-induced hypertension, but WT females were protected (124.3 ± 5.15 vs. 95.37 ± 4.882 mmHg, p=0.0114). Moreover, W521R male mutants had elevated mean arterial pressure in response to 3 weeks of high salt-feeding (4% NaCl) compared to the salt-sensitive WT littermate males (169.3 ± 27.69 vs. 124.3 ± 5.15 mmHg, p= 0.0024). Similar to WT females, female W521R mice were protected from salt-induced blood pressure elevation (102.3± 5.23 mmHg). Using flow cytometry, we found that high Na
+
increased dendritic cell IsoLGs production in male W521R mice compared to littermate controls (3.97 ± 1.68% vs. 1.01 ± 0.08 % of dendritic cells, p=0.0386). Our findings suggest that W521R gain-of-function mutation contributes to inflammation and hypertension in male but not female mice.
Abstract 080: SEX AND GENDER ASPECTS IN HYPERTENSION PREVALENCE OF CANADIAN AND EUROPEAN POPULATIONS
Authors: Azizi Zahra , Lindner Simon , Macías Ruiz María del Carmen , Alipour Pouria , Raparelli Valeria , Norris Colleen M , Kublickiene Karolina , Kautzky Willer Alexandra , Klimek Peter , El Emam Khaled , Fernández Villalba Emiliano , Herrero Maria-Trinidad , Pilote Louise , Investigators The GOING-FWD
Published: September 2022

Introduction:
Gendered-psycho-socio-cultural factors have been shown to play a significant role in disease manifestation, control and management of hypertension (HTN), and their relationship varies in males and females. We investigated the role of sex and gender in HTN prevalence and country-level differences in Canadian and European populations.

Methods:
Data from the Canadian Community Health Survey (CCHS, 2015-16, N=109,659, Females:56.6%) and the European Health Interview Survey (E-HIS, 2013-2015, N=316,333, females: 51.3%) were analyzed. Primary endpoint was defined as having a diagnosed HTN made by a health professional in the past 12 months. Relationship between gender variables and HTN prevalence and interaction with sex was assessed in a multivariable model. Federated analysis was conducted using the R package and DataShield which allows international data pooling by only exposing aggregated results.

Results:
The prevalence of HTN was greater in Canada compared with Europe (CCHS: 30.1% vs EHIS: 22.4%, P<0.001). Amongst European countries, Southern (SEU) and Central East (CEU) region had greater prevalence of HTN and more significant sex-differences (greater prevalence in females) compared to Northern (NEU) and Western (WEU) regions. In the multivariable model for assessing the role of gender variables in prevalence of HTN, female sex, older age, greater BMI, married or divorced/widowed status, and lower income were associated with higher risk of HTN, while greater household size, higher level of education, and living in European countries compared to Canada were associated with lower risk. There was a significant interaction between socioeconomic status (income, education) and sex in country stratified analysis. Within European countries, this was more evident in CEU, and SEU compared to NEU and WEU, where women with lower socioeconomic status had greater risk of HTN. There were significant country-level differences with being an immigrant and risk of having HTN. While living in NEU and SEU was associated with lower risk of having HTN, living in CEU was associated with a greater risk.

Conclusion:
The findings of the study demonstrate the importance of gender related factors and particularly the differences amongst various countries.
Abstract 081: Pulse Wave Velocity And Blood Pressure Variability As Prognostic Indicators In Very Elderly Patients With Advanced Chronic Conditions
Authors: De La Sierra Alejandro , Sierra Cristina , Murillo Marcos , Aiello Tommaso-Francesco , Mateu Aina , Almagro Pedro
Published: September 2022

The prognostic value of BP elevation in very old patients with other chronic comorbidities is uncertain. We aimed to assess the prognostic impact of 24-hour BP (both brachial and central), BP variability, and pulse wave velocity (PWV) in very elderly patients hospitalized due to a chronic disease decompensation. We included 249 patients older than 80 years, admitted to the hospital due to decompensation of congestive heart failure (149), chronic obstructive pulmonary disease (60), chronic kidney disease with acute kidney injury (26), or other decompensated chronic conditions (14). During hospital stay, 24-h BP monitoring was performed (Mobil-O-Graph PWV). Mean values of brachial and central BP, aortic PWV, and BP variability (24-hour SD) were obtained in all participants. After discharge, patients were followed-up for one year. The primary outcome was total mortality. Hazard ratio (HR) of BP estimates (for 1 SD increase) were obtained through Cox models, adjusted for clinical confounders. During follow-up 72 patients (29%) died. Progression of heart failure (58%) was the most frequent cause of death. No differences were observed in 24-h BP between those who died or remained alive. Systolic BP-SD, and PWV were higher in patients who subsequently died, while 24-hour HR- SD was lower (Table). In fully adjusted models, PWV (HR: 3.54; 95%CI: 2.37-5.28), SD of 24-h brachial systolic BP (1.29; 1.00-1.67) and SD of 24-h heart rate (0.65; 0.46-0.91) were associated with the risk of mortality. We conclude that PWV, SBP and heart rate variabilities, but not the level of BP are associated with mortality in very old patients with advanced chronic conditions
Abstract 083: State Of Ambulatory Hypertension Education In U.S. Internal Medicine Residency Programs: A Needs Assessment Survey
Authors: Gallagher Benjamin D , Windish Donna
Published: September 2022

Background:
Hypertension is one of the most common chronic conditions encountered by internal medicine residents during their ambulatory training. However, little is known about the current state of hypertension education for residents.

Objective:
To determine internal medicine residents’ confidence and training in key hypertension topics, desire for future training, and preferred teaching methods for learning about hypertension.

Methods:
We developed our survey by reviewing the literature, seeking input from two content experts, and piloting questions with chief residents at our institution. From November 2021 to March 2022 we administered the survey to internal medicine residents attending scheduled didactic sessions at 14 programs in different U.S. regions. Surveys were completed online or on paper, participation was voluntary, and responses were anonymous. The survey asked residents to rate their confidence, on a 4-point Likert scale, in 19 hypertension topics, and inquired about the type of training they had received in these topics. Residents were also asked to list the top five hypertension topics they wanted to learn more about and to list their top three preferred teaching methods for future training.

Results:
Two-hundred eleven of 324 residents (65%) responded to the survey. Many reported lacking training (25-40%) and confidence (52-81%) in key topics (Table).

Conclusions:
U.S. internal medicine residents in our study report insufficient training on key aspects of hypertension management. They describe low confidence in these areas and desire to learn more about them. Further curriculum development is needed to improve residents’ hypertension knowledge.
Abstract 084: Growth Stress And Cardiorenal Syndrome: Effect Of Renal Insufficiency On Blood Pressure And Cardiac Remodeling In Juvenile Rats
Authors: Kriegel Alison J , Patil Chetan N , Schwasinger Emma
Published: September 2022

Pediatric patients living with chronic kidney disease (CKD) can experience many of the comorbidities associated with CKD in adults including hypertension and cardiovascular (CV) pathology, as well as impaired somatic growth. The goal of this study was to understand how CKD impacts blood pressure and cardiac remodeling during a period of rapid somatic growth. We hypothesized that renal insufficiency in juvenile rats would lead to both impaired growth and cardiac hypertrophy, demonstrating that it is an appropriate model for studying mechanisms of cardiorenal syndrome (CRS) in pediatric patients. Young male Sprague Dawley rats (4-5 weeks of age) were weighed and underwent nuclear magnetic resonance imaging to assess body mass and composition and were then randomized to naïve (n=6), Sham (n=9) or 5/6 nephrectomy (5/6Nx; n=11) groups. Rats underwent echocardiography before surgery and at the end of the 4 week study. Anesthetized blood pressure was measured via the carotid artery. Tissues were collected weighed and stored for histological and molecular analysis. Results presented as mean±SEM, one-way ANOVA and p<0.05. There were no differences in body weights or composition between groups at the start of the study. No differences were measured between naïve and sham groups throughout the study. The 5/6Nx rats experienced less somatic growth than sham rats. Final body weight was lower in 5/6Nx rats than sham rats (229.2±4.0 vs. 229.6±17g). Similarly, tibialis length (TL) was smaller in 5/6Nx rats than sham rats (35.97±0.64 vs. 38.32±0.22 mm). The 5/6Nx group experienced a large increase in SBP when compared to sham animals (195.2±6.54 vs. 123.8±4.53 mmHg). Heart weight was higher in 5/6Nx vs. sham rats, despite the smaller body size. This difference was even more apparent when heart weight was normalized to TL (5/6Nx, 0.34± 0.10 vs. sham, 0.27±0.01 g/cm). These studies indicate that 5/6Nx surgery in juvenile SD rats can recapitulate characteristics of pediatric CRS and CKD-related growth impairment. Adult male 5/6Nx SD rats in our prior studies developed cardiac hypertrophy in the absence of hypertension. These results suggest that the added stress of growth during renal insufficiency can drive both hypertension and cardiac hypertrophy in juvenile rats.
Abstract 085: Pediatric Dyslipidemia Is Associated With Higher Urinary Angiotensin Converting Enzyme Activity And Blood Pressure Values In A Brazilian Cohort
Authors: Azinheira Nobrega Cruz Nayara , Oliveira Lilian Caroline G , MILL JOSE GERALDO , Casarini Dulce E
Published: September 2022

Angiotensin converting enzyme (ACE) upregulation in adults contributes to cardiometabolic diseases including dyslipidemia and atherosclerosis.

Objective:
Investigate whether ACE activity is modulated in pediatric dyslipidemia.

Methods:
We evaluated anthropometric measurements, blood pressure and fasting lipid concentrations of 360 individuals (190 boys and 170 girls) aged from 6 to 19 year (mean 11.6). Categorization was done according to the levels of each lipoprotein (Total cholesterol (TC), triglycerides (TG), LDL-C, HDL-C, and non-HDL) into three groups: normolipidemic, borderline and dyslipidemic; reference values were according to American Academy of Pediatrics and Brazilian Pediatric Society. ACE activity in urine was measured with the substrates Z-FHL-OH and hippuryl-HL-OH. ACE activity ratio to infer N-domain activity was calculated (Z-FHL/h-HL).

Results:
Dyslipidemic levels of HDL-C, TG and LDL-C were observed in 23%, 9% and 3% of the participants, respectively. These clinical alterations were more frequent in obese children (Chi-square, p <0.001). ACE activity (Z-FHL) was higher in the groups with borderline and dyslipidemic values of HDL-C than in normolipidemic group (0.077 vs 0.070 vs 0.037 nmol/min/mg of creatinine, p = 0.01). The ratio of ACE activity was augmented in HDL-C borderline group when compared to HDL-C normolipidemic group (5.06 vs 2.39, p <0.01) and in LDL-C dyslipidemic group than in LDL-C borderline and LDL-C normolipidemic groups (8.66 vs 1.84 vs 2.88, p = 0.02). Volunteers with normal levels of TG presented lower diastolic blood pressure (DBP) mmHg (p = 0.02), percentile of DBP (p < 0.01), and percentile of systolic blood pressure (SBP) (p <0.01) than volunteers with borderline and dyslipidemic levels of TG. Also, increased DBP mmHg was observed in borderline and dyslipidemic HDL-C groups when compared to normolipidic HDL-C group.

Conclusion:
Pediatric hypertriglyceridemia, HDL-C deficiency and high LDL-C were related to higher urinary ACE activity (mainly N-domain). ACE modulation can contribute to higher risk of hypertension and other cardiometabolic alterations still in childhood and adolescence. Also, impairment in lipoprotein levels was associated with higher blood pressure values.
Abstract 104: Kisspeptin Signaling And Pregnancy Outcomes Are Ameliorated In The Preeclamptic-like BPH/5 Mouse After Synchronization Of Sex Steroid Hormones
Authors: Gomes Viviane C , Woods Ashley K , Landry Camille A , Crissman Kassandra R , Sones Jennifer L
Published: September 2022

Kisspeptins (KP), encoded by
Kiss1
, are reported to play a role in early-onset preeclampsia (ePE), a life-threatening hypertensive disorder of pregnancy. KP is higher in term ePE placentae than in normal pregnancies. However, the role of KP in PE has not been confirmed. The Blood Pressure High Subline 5 (BPH/5) mouse model spontaneously develops the main features of ePE, including impaired placental expansion and maternal hypertension. We have shown that
Kiss1
is higher at the BPH/5 maternal-fetal interface on embryonic days (e) 4.5 and 7.5. BPH/5 females have abnormal circulating sex steroid hormones (SSH) during early gestation (e2.5), with lower estradiol-17ß (E2) and higher progesterone (P4). Since E2 and P4 may modulate uterine
Kiss1
, we aimed to investigate the effects of artificial synchronization of SSH (AS-SSH) in BPH/5 uteroplacental KP signaling and placental development. We hypothesized that AS-SSH would prevent BPH/5 uteroplacental
Kiss1
upregulation and normalize KP downstream expression of tissue inhibitor of metalloproteinase (TIMP) 2 and placental defects. AS-SSH was performed after ovariectomy of pregnant BPH/5 and C57 females (n = 8/strain) at e2.5 with one dose of E2 (25 ng/mouse, subcutaneously), and daily subcutaneous injections of P4 (1ng/mouse) until embryonic implantation sites (eIS) were collected 3 days post-surgery (n = 8/group). Contrary to natural (Nat) BPH/5 pregnancies, eIS
Kiss1
and
TIMP2
expression was not different between AS-SSH BPH/5 and C57 (p > 0.05). Nat and AS-SSH pregnant mice were anesthetized at e12.5 to assess umbilical cord blood flow via ultrasound and uteroplacental units were harvested for placental morphometry (n = 4-6/group). Placental expansion into the maternal decidua was measured and expressed as the ratio of the placenta depth in relation to the placenta + decidua. Nat BPH/5 had lower placental expansion and umbilical blood flow than Nat C57, whereas AS-SSH BPH/5 had higher placental expansion and umbilical blood flow than Nat BPH/5 (p < 0.05). In conclusion, synchronization of the E2 and P4 profile in the BPH/5 mouse early gestation mitigated
Kiss1
and
TIMP2
upregulation at the maternal-fetal interface and ameliorated the placental defects previously reported in this model.
Abstract 110: Cullin3 Deficiency In T Lymphocytes Causes A Lean Phenotype With Increased Carbohydrate Catabolism But Promotes Hypertension And Renal Inflammation
Authors: Wu Jing , Lu Ko-Ting , Reho John J , Wackman Kelsey , Liu Jing , Grobe Justin L , Singer Jeffrey , Sigmund Curt D
Published: September 2022

Low body mass is associated with increased risk of death and stroke, but the mechanism is unclear. Cullin3 (Cul3) ubiquitin ligase regulates T lymphocyte polarization, insulin signaling, and blood pressure (BP) and may provide a mechanistic link for the cardiometabolic phenotypes. Deletion of Cul3 in T cells (

Cul3
fl/fl

x
CD4-Cre
, termed

CD4-Cul3
-/-

) resulted in 17% lower body weight (BW:

CD4-Cul3
-/-
vs CD4-Cul3
+/+

littermates, 23.9 ± 0.9 vs 28.7 ± 0.7 g, 12-16 wks old, n = 6, 12, p<0.01) and blunted growth curve (ΔBW over 7 wks: 3.2 ± 0.9 vs 5.0 ± 0.6 g) in male mice. No difference in BW or growth curve was observed in females. Nuclear magnetic resonance analysis found decreased fat mass (6.4 ± 0.4 vs 9.2 ± 0.8 g, n = 10, 13, p < 0.01) in male null mice with no change in lean mass or fluid content. This was confirmed by post-mortem quantitation of inguinal fat and interscapular brown adipose tissue. There was no difference in postprandial blood glucose, but the male null mice exhibited lower area under curve (AUC mg/dL x hr, 543 ± 104 vs 765 ± 40, p < 0.05) in glucose tolerance test (2 g/kg, 5 hr fasting) and developed hypoglycemia and seizure during fasting insulin tolerance test (0.5 U/kg).

CD4-Cul3
-/-

mice had elevated respiratory exchange ratio (ratio between CO
2
produced by the body and O
2
consumed) (0.83 ± 0.01 vs 0.78 ± 0.01, n= 4, p < 0.05), suggesting a larger reliance on carbohydrates rather than lipids as fuel. Baseline BP was comparable in null and control mice. In response to chronic pressor dose of angiotensin (Ang) II (490 ng/kg/min, s.c. 14 days),

CD4-Cul3
-/-

mice displayed exaggerated hypertension (systolic BP in mmHg: 164 ± 8.4 vs 150.3 ± 4.9; ΔSBP: 35.3 ± 7.5 vs 15.8 ± 5.3, p < 0.05). Mean and diastolic BP were also greater in the null mice, but there was no difference in heart rate or activity. Ang II-infused

CD4-Cul3
-/-

mice showed a trend towards increased renal CD3
+
T cells and significantly increased CD4
+
T helper cells (p < 0.05). Thus, T cell Cul3 deficiency caused a lean phenotype characterized by decreased adiposity, increased carbohydrate catabolism, and augmented Ang II-induced hypertension and renal inflammation. The

CD4-Cul3
-/-

mice may be a useful model to study the association between low body mass and increased cardiovascular risk in humans.