bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Features and Risk Factors for Outcome in Hemodialysis Patients with COVID-19 after Complete Liberalization of Epidemic Control in China
Authors: Zhang Shuang , Liu Shu-Xin , Wang Zhi-Hong , Xiao Ping , Liu Hong , Lu Yan , Dong Cui , You Lian-Lian
Published: October 2024

Introduction: Patients undergoing hemodialysis (HD) are highly vulnerable during the COVID-19 pandemic. We aimed to investigate the risk factors associated with the severity of COVID-19 and death after the complete liberalization of epidemic control in China. Methods: We followed the outcomes of the HD patients of Central Hospital of Dalian University of Technology, from December 6, 2022, to January 8, 2023. The non-contrast-enhanced chest computed tomography (CT) was performed on all COVID-19-infected hospitalized patients. We recorded the patient’s clinical characteristics, demographic features, vaccination history, treatments, and lung lesions. Odds ratios and 95% confidence intervals were calculated using logistic regression models to identify independent risk factors for COVID-19-related severity and mortality. Results: This study included a total of 858 HD patients, of which 660 were infected with COVID-19. The mean age was (55.61 ± 14.61) years, with a median (interquartile range) dialysis duration of 44.5 (69.5) months. Over half (60%) of the study participants were male, and the majority had hypertension as a comorbidity. Multivariable analysis revealed that age, pre-dialysis diastolic pressure, fever, white blood cell (WBC) count, potassium, β2-microglobulin level, and calcium were independent risk factors for disease severity, while platelets, urea nitrogen, serum chlorine and creatinine were identified as independent protective factors. Furthermore, total iron-binding capacity and vaccination were found to be independent protective factors against mortality, and WBC count was an independent risk factor for in-hospital mortality (p < 0.05). The most frequent CT finding among hospitalized patients with chest symptoms was patchy shadow or pleural effusion, observed in 64.8% of cases. More than half of the patients exhibited bilateral lung lesions, and over 60% involved two or more lobes. Conclusion: The majority of HD patients are susceptible to COVID-19. Demographic, clinical features, and laboratory indicators can be used to predict the severity and mortality associated with COVID-19. Our findings will assist clinicians in identifying markers for the early detection of high mortality risk in HD patients with COVID-19.
The Relationship between Early Exposure to General Anesthesia and Neurobehavioral Deficits
Authors: Xu Jinnan , Wang Fan , Gao Yi , Qi Chuanyu , Chen Tiannan , Yan Jia
Published: October 2024

Background: In contemporary medical practice, general anesthesia plays an essential role in pediatric surgical procedures. While modern anesthetic protocols have demonstrated safety and efficacy across various pathological conditions, concerns persist regarding the potential neurotoxic effects associated with early exposure to general anesthesia. Summary: Current research primarily examines the neurocognitive developmental impacts, with limited focus on neurobehavioral developmental disorders. This review presents a comprehensive analysis of clinical trial results related to five critical neurobehavioral developmental disorders: fine motor disability, attention-deficit hyperactivity disorder, impulse control disorders, autism spectrum disorder, and developmental coordination disorder. Furthermore, this review synthesizes insights from basic research on the potential toxicological mechanisms of general anesthetic agents that could influence clinical neurobehavioral changes. These findings provide valuable guidance for the prudent and safe utilization of anesthetic agents in pediatric patients. Key Messages: This review explores the potential connections between general anesthesia and five neurobehavioral disorders, highlighting the importance of cautious anesthetic use in children in light of current research findings.
Anti-inflamatórios não esteroidais (AINEs) x microbiota: uma simbiose ou disbiose intestinal?
Authors: Mello Danilo Saragiotto Ferreira de , Mendes Anderson Benegas , Tanimaru Aline Namie , Gonzalez Fabiana Gaspar , Carvalho Celine de Furtado
Published: October 2024

Background: Considering the widespread use of non-steroidal anti-inflammatory drugs (NSAIDs), the following study aimed to discuss their potential effects on the intestinal microbiota and the consequent repercussions in clinical practice. Materials and methods: The development of the bibliographic review was based on the “PubMed” research platform, through the use of the keywords: “NSAIDs”/“Non-steroidal anti-inflammatory drugs” and “Microbiota”/“Microbiome”/“Gut microbiota”, with the full reading of those articles published in the last 5 years in Portuguese, English or Spanish. Meta-analyses, systematic reviews and other bibliographic reviews were excluded from the study, resulting in a final sample of 14 articles. Results: The evidence of quantitative and qualitative changes in the intestinal microbiome attributed to the use of these drugs was significant, with an increase in Gram-negative microorganisms and a reduction in Gram-positive microorganisms. However, the altered microorganisms, the observed pathologies, and other factors involved, as well as the intervention strategies addressed to treat the dysbiosis, do not yet have their mechanisms completely defined. Conclusion: Despite the inconclusive results, a therapeutic alternative for the presented dysbiosis would be the use of probiotics from Gram-positive strains. In addition, room is opened for the benefit of new studies in the area
Abstract 01: Spatial Transcriptomics Reveals How Smooth Muscle Cells Transform to Renin-Expressing Cells During Homeostatic Challenge
Authors: Smith Jason , Yamaguchi Manako , Sequeira Maria Luisa , Gomez Roberto
Published: October 2024

Introduction:
Juxtaglomerular (JG) cells sense changes in blood pressure through communication with neighboring cells, and/or the sympathetic nervous system upon which they release renin to maintain homeostasis. To meet the demands for circulating renin when homeostasis is threatened, smooth muscle cells (SMCs) of the renal arterioles are transformed to renin-synthesizing cells. Discriminating transformed cells versus JG cells using sorting and scRNA-seq is cumbersome, limiting our ability to explore the events underlying such transformation. Therefore, there is a need to study these cells
in situ
to assign and determine which epigenomic and cell communication pathways regulate renin expression and SMC transformation.

Hypothesis:
Transformation of SMCs to renin-expressing cells during homeostatic stress requires specific signals from neighboring cells in addition to intrinsic transcriptomic changes.

Methods:
A pilot study was performed using two three-month-old C57BL/6 mice fed either a normal diet, or a low-salt diet with the RAS inhibitor, captopril, to induce homeostatic stress. After one week, the kidneys were harvested and prepared for the 10X Genomics Visium platform. Sequencing results were preprocessed using the Space Ranger pipeline (v. 2.0.1) with the mouse mm10 genome prior to downstream analysis using the R packages Seurat, Signac, and CellChat.

Results:
Homeostatic challenge significantly increased
Ren1
expression (
p
Holm-adj.
=4.63x10
-10
) and led to more and stronger cellular interactions. Interstitial cells and JG cells contributed the largest percent increase in the observed signaling changes (6.5% and 4.5% respectively). Increased signals for collagen, amyloid precursor protein, osteopontin, laminin, thrombospondin 1, vascular endothelial growth factor, and growth arrest specific pathways drove changes between native and transformed renin-expressing cells. In addition to the established JG cell markers
Ren1
and
Akr1b7
, transformed cells had significantly increased expression (
p
Bonferroni-adj
< 0.01) of
Hmgcs2
,
Cyp4a14
,
Suox
,
Acta2
,
Slc2a5
,
Ptp4a3
,
Sncg
, and
Rad21
.

Conclusions:
In conclusion, spatial transcriptomics revealed the evolution of SMC transformation into renin phenotypes by uncovering pathways and genes understudied in the context of renin regulation. Future work will leverage higher resolution spatial technology and further explore the contribution of identified pathways to renin regulation and SMC transformation.
Abstract 03: Glomerular Hyperfiltration Promotes Chronic Kidney Disease through Interleukin 17A-Mediated Inflammation
Authors: Liu Jing , Chen Luojing , Le Thu , Wu Jing
Published: October 2024

Impaired renal autoregulation in obesity, diabetes and hypertension (HTN) causes elevated glomerular capillary pressure and glomerular hyperfiltration, initiating progressive glomerulosclerosis (GS), nephron loss and chronic kidney disease (CKD). Increased renal perfusion pressure drives renal T cell infiltration. T helper (Th) 17 cells promote vascular stiffening in HTN and renal interstitial fibrosis post AKI, but their roles in GS and CKD are poorly understood. We hypothesized that glomerular hyperfiltration promotes CKD through Th17-mediated GS and tested the hypothesis in 129S6 mice using the reduced kidney mass model (RKM) induced by uninephrectomy and partial renal artery ligation in the remaining kidney (functional renal mass reduced by ~ 5/6). BP (radiotelemetry) and GFR (FITC-sinistrin) of RKM mice were identical to sham controls at baseline (BL, n=4-6). Immediately after the RKM procedure, GFR dropped to 22% of BL, consistent with the extent of renal mass ablation. Mean BP increased by 29 mmHg first week after 5/6 ablation (144 ± 8 vs BL 112 ± 2 mmHg, p<0.05, 2-way ANOVA) and remained elevated through week 12, exposing the remnant nephrons to increased renal perfusion pressure. Serum creatinine, BUN and urinary albumin were significantly elevated after 5/6 ablation. In RKM mice, GFR gradually increased to 38% of BL at day 3, 57% at week 1 and 68% at week 2, suggesting that systemic HTN and impaired renal autoregulation may have caused substantial single-nephron hyperfiltration in the remnant kidney. GFR plateaued at ~ 70% of BL in week 3-4 then declined to 48% at week 8 when glomerular, interstitial, and microvascular fibrosis developed as evidence by Periodic Acid Schiff and Picro Sirius Red staining. Importantly, the early adaptive increase of GFR in the first 2 weeks post 5/6 ablation was accompanied by a striking increase of renal interleukin (IL)-17A+ T cells, which preceded the late infiltration of CD11b+ myeloid cells, F4/80+ monocyte/macrophages, and T regulatory cells. Anti-IL17A antibodies (eBioMM17F3, 100 μg i.p. twice weekly) attenuated the early hyperfiltration, abolished aortic/renal inflammation, and prevented GFR decline and GS at week 8 in male but not female RKM mice, likely due to the anti-hypertensive and anti-inflammatory effects of IL-17A neutralization. We conclude that glomerular hyperfiltration causes progressive nephron loss and kidney fibrosis in CKD, at least in part, through IL-17A mediated renal inflammation.
Abstract 05: Renal Resident Tissue Macrophages Contribute to Salt-Dependent Increases in IL-6
Authors: Cowley Craig , Fairbanks Landon , Yoshigi Masaaki , Araos Patricio , Beaudin Anna , Wynne Brandi
Published: October 2024

Salt-Sensitive hypertension is associated with an increase in pro-inflammatory cytokines and immune cells. We previously showed an acute increase in systolic blood pressure following IL-6 + high salt (HS) after 3 days, coupled with an immediate (1 day) reduction in sodium excretion. Renal resident tissue macrophages (rRTMs) form an intricate network within the kidney and function to both maintain homeostasis and secrete pro-inflammatory cytokines. We hypothesize that rRTMs are primary contributors to the immediate increases in IL-6 following high salt.

To test this hypothesis, male wild-type mice lacking a functional CX
3
CR
1
chemokine receptor (CX
3
CR
1
-
EGFP+/+
) or wild-type (Wt, C57Bl6) were used. CX
3
CR
1
is required for RTM localization to the kidney; thus, these mice are rRTM-depleted. Mice were fed HS (1% saline or 4% HS chow) or normal chow (NC) for 6 days and kidneys harvested. Kidneys were digested and FACS analysis performed on total immune cell populations or used for RT-PCR (n=4-5 Wt & rRTM-depleted). Cells were gated on: Live + CD45
+
+ MHCII + Lyc6C
-
+ B220
-
+ CD64
+
+ F4/F80
hi
+ CD11b
lo.
Or Lyc6C
hi
. Lyc6C
hi
and rRTMs were pre-stimulated (LPS, 1mg/mL, 3hrs) before staining with intracellular IL-6 antibody. FACS data were analyzed as a percent increase (mean fluorescence intensity, MFI) of IL-6 over control. RT-PCR data were analyzed as DCt/housekeeping.

Our data show that rRTMs are the most abundant immune cell in the kidney (23.4% of total CD45
+
) and predominantly reside in the cortex and outer medulla (>99%). HS feeding (4%) significantly increased whole kidney IL-6 mRNA (12.0 ± 0.5
vs
. 15.3 ± 0.6 DCt Wt NC, **p<0.01). This increase was completely prevented in rRTM-depleted mice. While intracellular IL-6 levels were generally increased in LPS-stimulated Lyc6C
hi
inflammatory monocytes, the rRTMs showed a preferential increase in intracellular IL-6 with both hyperosmolar saline (1%, >13% increase) and HS chow (4%, >50%).

Here, we show that rRTM-depletion prevents the HS-induced increase in kidney IL-6. We additionally show that rRTMs are preferentially increasing intracellular levels of IL-6 following an acute HS load of hyperosmolar saline (1%) or HS chow (4%). We previously demonstrated that IL-6 contributes to increased blood pressure and sodium retention, and these data suggest that rRTMs preferentially contribute to the increased IL-6.
Abstract 06: Role of the Atrial Natriuretic Peptide in Mitochondria-mediated Proximal Tubule Epithelial Oxidative Stress in Type 1 Diabetic Kidney Disease
Authors: Spires Denisha , Cherezova Alena , Palygin Oleg , Ilatovskaya Daria
Published: October 2024

Introduction:
Diabetic kidney disease (DKD) is the leading cause of end-stage renal disease. Most studies of DKD are glomerulo-centric. The recent progress in the control of renal glucose handling by SGLT2i makes the proximal tubule (PT) a prime candidate. Mitochondrial (mito) dysfunction and mito-induced oxidative stress in DKD is well-studied and is known to accelerate tubular injury. Some studies have suggested the role of the Atrial Natriuretic Peptide (ANP) in DKD. ANP is a hormone that regulates renal salt/water homeostasis and renal hemodynamics. We hypothesize that in DKD, ANP is a beneficial factor that improves proximal tubule mitochondrial function via protection from oxidative stress-induced ferroptosis.

Methods:
Using male ANP knock-out (SS
NPPA-/-
) and wild-type (SS
WT
) rats on the Dahl SS background, T1DKD was induced by streptozotocin (STZ; 75mg/kg) at 9-10 weeks of age. Baseline, midpoint, and endpoint blood glucose, urine, and body weights were collected. Western blotting was done on isolated cortical tissue and tissue fibrosis was measured in PicroSirius Red staining. ANOVA through Origin 2019b was used for comparisons.

Results:
STZ- SS
WT
and STZ-SS
NPPA-/-
groups exhibited similar body weight (231.7 ± 14.8 vs. 230.9 ± 8.4 g, respectively), two-kidney weight to body weight ratio (31.8 ± 0.9 vs. 28.4 ± 1.5 mg/g), and endpoint hyperglycemia (with blood glucose levels >650 mg/dL). Both, STZ- SS
WT
and STZ-SS
NPPA-/-
groups had high diuresis (59.9 ± 3.1 vs. 52.7 ± 3.2 mL/24hr/100gTBW). Histological analysis revealed elevated cortical fibrosis in the STZ-SS
NPPA-/-
compared to STZ- SS
WT
(4.6 ± 0.2 vs. 3.1 ± 0.2 % area, respectively, p<0.05). Moreover, in the kidney cortex, STZ-SS
NPPA-/-
rats compared to STZ- SS
WT
rats exhibit a significant increase in OXPHOS complexes IV and V (p<0.05) and a decrease in glutathione peroxidase 4 (GPX4; a core regulator of ferroptosis) abundance (1.5 ± 0.2 vs. 1.9 ± 0.1 a.u., respectively, p<0.05).

Conclusion:
Data showed that lack of ANP may promote DKD-related tubulointerstitial fibrosis and suggested a relationship with renal cortical mitochondrial function. Future studies will provide mechanistic insight into the role of ANP in mito-related oxidative stress and further assess the role of ANP in PT ferroptosis.
Abstract 08: Genome-wide Maps of Human Kidney Proximal Tubules and Thick Ascending Limbs of Henle reveal an Enrichment for Human Blood Pressure associated Polymorphisms
Authors: Ray Atrayee , Yang Chun , Stelloh Cary , Tutaj Monika , Liu Pengyuan , Liu Yong , Lin Chien-Wei , Widlansky Michael , Kwitek Anne , Geurts Aron , Cowley Allen , Liang Mingyu , Greene Andrew , Rao Sridhar
Published: October 2024

GWAS has identified >1,000 blood pressure (BP)-associated sentinel SNPs, most of which (>90%) are intronic and intergenic. How they regulate BP-relevant genes remains an open question. We hypothesized many SNPs would reside within
cis
-regulatory elements specific to nephron segments critical to sodium resorption such as proximal tubule (PT) and thick ascending limb (TAL). To address this, we developed chromatin state (ATAC-seq) and transcriptome (RNA-seq) maps from manually dissected human PT and medullary TAL (mTAL) segments. We identified 216,168 and 179,467 open regions in PT and mTAL, respectively, with the majority (70%) being unique to one tissue. Interestingly, peaks shared between the two segments resided more frequently in promoters, whereas unique peaks were more often found in distal introns or intergenic regions, suggesting they lie within enhancers (
Figure 1A
). Both chromatin state and transcriptomic data demonstrated a correlation between open chromatin peaks within gene promoters and mRNA levels (r=0.11; p<4.6e-16). This correlation was weaker at open regions distal to the promoter, illustrating the difficulty of mapping distal open regions to the gene(s) they regulate.

We hypothesized these open regions would be enriched for BP-relevant SNPs. We curated 1,071 bp-associated SNPs and found 87 and 67 in mTAL and PT, open regions respectively, many of which are near BP-relevant genes. This enrichment was significant based on 10
4
randomizations (pValue 10
-4
) (
Figure 1B
). We conclude that these novel tissue-specific chromatin maps can be used to identify potential mechanisms by which SNPs influence gene expression and BP. Additional studies will be needed to demonstrate both causality and molecular mechanisms.
Abstract 12: Deficiency of Progranulin Decreases Mitochondrial Function and Induces a Whitening Phenotype in Perivascular Adipose Tissue, Suppressing Its Anti-Contractile Effects
Authors: Costa Rafael , Bruder Nascimento Ariane , Richert Adam , Goetzman Eric , Bruder Thiago
Published: October 2024

Perivascular adipose tissue (PVAT) is recognized as an endocrine organ that secretes a variety of adipokines, which negatively regulate vascular contraction. Progranulin (PGRN), a glycoprotein encoded by the GRN gene, is highly expressed in adipocytes, and is implicated in inflammation, cellular proliferation, and repair mechanisms. The relationship between PGRN and PVAT remains unclear. We hypothesized that the lack of PGRN negatively affects PVAT quality. We used male C57BL/6J wild-type (WT) and global PGRN knockout (KO) mice, aged 10-12 weeks. Energy expenditure was analyzed via the Comprehensive Lab Animal Monitoring System (CLAMS). Vascular function was studied in endothelium-intact aortae with PVAT [PVAT(+)] and without PVAT [PVAT(-)]. Additionally, 3T3-L1 cells were used to evaluate the molecular mechanisms. The KO mice showed a decreased respiratory exchange ratio (RER) and a whitening phenotype of PVAT, which was accompanied by a loss of anti-contractile effects [(mN) WT_PVAT(-): 5.2±0.3 vs PVAT(+): 3.4±0.5*, *P<0.05 and KO_PVAT(-): 6.1±0.7 vs KO_PVAT(+): 5.7±0.4]. Additionally, PVAT from KO displayed a significant reduction in nitric oxide production [(RFU) WT_PVAT: 223.1± 12.3 vs KO_PVAT: 67.3±9.1*, *P<0.05] and endothelial NO synthase phosphorylation. Additionally, the lack of PGRN attenuated the expression of mitochondrial complexes [(AU) WT_PVAT: 1.3± 0.03 vs KO_PVAT: 0.6±0.06*, *P<0.05] measured by the OXPHOS cocktail antibody. Via high-resolution respirometry, we found that isolated mitochondria from KO_PVAT showed impaired succinate-driven respiration or complex II activity. To understand if the whitening phenotype of PVAT was leading to its dysfunction, we induced a browning phenotype of PVAT by treating the mice with a β3-agonist for 7 days (CL 316243, 1mg/kg/day). This treatment increased RER restored the PVAT phenotype, and improved PVAT anti-contractile effect and mitochondrial respiration. Using 3T3-L1 cells, we engineered the overexpression and knockdown of PGRN via lentivirus (LV) infection and siRNA, respectively. Suppression of PGRN increased mRNA of whitening markers (1.5-fold increase vs scramble siRNA), while overexpression elevated mRNA of browning and mitochondrial markers (1.3-fold increase vs scramble LV). In summary, our findings reveal that PGRN is crucial for maintaining the phenotype and anti-contractile function of PVAT and highlight PGRN as a pharmacological target for vascular outcomes in cardiometabolic conditions.
Abstract 14: Arterial Stiffness in Obese Normotensive Children: The First Cardiovascular Risk Biomarker?
Authors: Duarte Mariano , Aquieri Analia , Rivas Silvia , Donth Leger Gimena , Pardo Valeria , Coyle Javier , Curia Andrea , Gonzalez Ballerga Esteban
Published: October 2024

Introduction:
Obesity is an emerging pandemic driven by consumption of a diet rich in fat, highly refined carbohydrates and a sedentary lifestyle in both children and adults. In obesity arterial stiffness (AS) often associated with endotelial dysfunction (ED) is an independent and strong predictor of cardiovascular disease (CVD) and is a precursor to atherosclerosis, systolic hypertension, cardiac diastolic dysfunction, and impairment of coronary and cerebral flow. In obese children, AS also promotes fatty liver disease whitch eventually leads to fibrosis.

Aim of Study:
In obese children (OC), central blood pressuere, arterial stiffness and endothelial dysfunction, liver steatosis/fibrosis were evaluated and compared with control group (CG), non-obese, non-hypertensive children.

Methods:
Carotid-femoral pulse wave velocity (PWV) is the current non-invasive method of choice to asses AS and PWV carotid-brachial variation pre- and post-induced transient ischemia of brachial artery to evaluate ED. Fibroscan (FS) and Control Attenuation Parameter (CAP) are a non-invasive reliable examination that allows to know the degree of liver steatosis and liver stiffness (fibrosis). Carotid-femoral PWV (AS) and PWV variation of brachial artery (ED) were assessed using Complior System® (Artech-Medical France). We evaluated 31 obese children between 10 and 13 years old (BMI 28.70±3.72) and compared to CG (between 11 and 13 years old, n=16, BMI 19.89±1.75)

Results:
In obese children (BMI 28.70±3.72 and Z score 2.02±0.42) vs CG (BMI 19.89±0.75 and Z core 0.57±0.24), PWV-CF (arterial stiffness) was significantly increased: 5.8±1.3 m/s vs. 3.4±0.9 m/s, p<0.01.

PWV-CB variation were paradogical (endothelial dysfunction) after transient ischemia in obese children (△+9% compared to baseline) and it was as epected in the CG (△-12.2% compared to baseline) (P<0.001). Fibroscan and CAP revealed that the degree of steatosis was significantly higher in OC but no differences were found in the degree of liver fibrosis.

Conclusions:
We found a significatly increase in arterial stiffness in obese children compared to CG. Arterial stiffness in obese children often precedes the development of hypertension and liver fibrosis (even in the presence of liver steatosis) suggesting that arterial stiffness is one of the earliest biomarkers for increased cardiovascular risk. Interventions aimed at attenuating obesity in chidhood are emerging as important target to improve public health outcomes.
Abstract 18: Persistent beneficial effects of angiotensinogen small interfering RNA (siRNA) and valsartan in spontaneously hypertensive rats after drug removal
Authors: Cruz-Lopez Edwyn , Ye Dien , Van veghel Richard , Garrelds Ingrid , Kasper Anne , Wassarman Kelly , Tu Ho-Chou , Zlatev Ivan , Danser Jan
Published: October 2024

Objective:
Dual blockade of the renin-angiotensin system (RAS) with siRNA targeting hepatic angiotensinogen (AGT) plus the angiotensin receptor blocker valsartan synergistically lowers blood pressure and diminishes cardiac hypertrophy in spontaneously hypertensive rats (SHR). Since the effects of one siRNA injection may last for up to six months, here we evaluated whether the REVERSIR (reverse siRNA silencing, RVR) technology and/or stopping valsartan might acutely reverse the effects of dual RAS blockade in SHR.

Methods:
Ten-week old SHR were subjected to a 3-week treatment with vehicle or AGT-siRNA (10 mg/kg) + valsartan (4 mg/kg per day), followed by administration of vehicle or AGT-RVR at doses of 1, 10 and 20 mg/kg, both with and without continuation of valsartan, for 1 week. Mean arterial blood pressure (MAP) was monitored using radiotelemetry, and circulating AGT and renin were measured by enzyme-kinetic assay.

Results:
Baseline MAP was 143±2 mm Hg. Dual treatment lowered MAP by ≈70 mm Hg, cardiac hypertrophy (heart weight/tibia length) by ≈30%, and circulating AGT by >99%, while renin increased >100-fold versus vehicle. Discontinuing valsartan or applying AGT-RVR (1, 10 or 20 mg/kg) increased MAP similarly (by ≈20 mm Hg), while discontinuing valsartan + AGT-RVR (10 mg/kg) increased MAP by ≈40 mm Hg. Changes in cardiac hypertrophy mimicked those in MAP. Only valsartan discontinuation + AGT-RVR fully restored circulating AGT and renin to normal. When discontinuing valsartan only, AGT remained suppressed by >95%, and renin was up 10-fold, while with AGT-RVR 1, 10 and 20 mg/kg only, AGT suppression was 25, 50 and 50%, while renin was up 25-, 15- and 15-fold, respectively.

Conclusions:
Discontinuing valsartan or applying AGT-RVR in SHR treated with valsartan + AGT siRNA partially restored MAP, AGT, and renin, likely reflecting the fact that this approach replaces dual RAS blockade by single RAS blockade. However, although discontinuing valsartan plus AGT-RVR fully restored RAS activity, MAP and cardiac hypertrophy remained suppressed under this condition. This agrees with observations that early RAS inhibition resets genetic pathways and networks, allowing persistent blood pressure normalization even when treatment has stopped.
Abstract 26: An Effective mHealth Intervention to Close the Guideline-to-Practice Gap in Hypertension Treatment: mGlide RCT
Authors: Lakshminarayan Kamakshi , Murray THOMAS , McCarthy Teresa , Luepker Russell , Drawz Paul , Streib Christopher , Everson-Rose Susan , Connett John , Westberg Sarah
Published: October 2024

Background:
Suboptimal HTN control is a widespread problem particularly among minorities and socioeconomically disadvantaged groups.

Objective:
To determine if a mobile health (mHealth) intervention facilitating an active partnership between inter-professional care teams (including pharmacists) and patients improves HTN control in patients with uncontrolled HTN.

Methods:
mGlide is an NIH-funded, 2-arm RCT evaluating HTN control between a mHealth intervention (mGlide) vs. state-of-clinical-care (comparison) groups using a PROBE (Prospective Randomized Open Blinded End-point) design. Table 1 shows intervention components. High CVD risk patients with uncontrolled HTN identified by EHR query were randomized 1:1 to either mGlide or to comparison for a 6-month intervention followed by a 6-month observation period. Recruitment sites include primary care clinics and specialty stroke service in an integrated health system, community clinics (CC) and federally qualified health centers (FQHC). The FQHC and CC serve low income and minority (Hmong and Latino) patients. The primary aim tested clinical effectiveness of mGlide in reducing SBP at 6 months. Secondary outcomes include resource utilization, adverse events and medication side-effects.

Results:
Recruitment started in 2019. N=395 participants enrolled by August 2023. The pandemic impacted enrollment; however enrollment recovered after vaccine availability (Figure). Primary outcome was available in March 2024 on everyone except in those lost to follow-up (n=38; 9.6%). We used multiple imputation for those missing 6 month SBP. Baseline mean SBP was 143.2 vs. 143.6 mm Hg in mGlide vs. comparison. The intervention led to better HTN control (Table 2). Mean 6-month SBP was 128.1 (mGlide) vs. 134 mm Hg (comparison). Adjusting for baseline SBP and study strata, 6-month mean SBP was 5.7 mm Hg (95% CI 2.9-8.4; p = 0.0001) lower in mGlide vs. comparison. The odds of HTN control defined as “alive and SBP < 130 mm Hg”, was 60% higher in the mGlide arm (OR 1.6; 95% CI 1.03-2.48; p=0.035). There was one death in each study arm. The mGlide arm had fewer ER visits (16% vs. 25%; p=0.024), fewer hospitalizations (7% vs. 13%; p=0.044) and more medication side effects (39% vs. 26%, p=0.008).

Conclusions:
An inexpensive mHealth intervention that used the patient’s own phone (Table 1) was implemented in diverse health systems and significantly reduced SBP and was associated with less resource utilization at 6 months (Table 2).
Abstract 32: Vγ6
+
γδ T cells protect against angiotensin II-induced hypertension and vascular injury in male mice
Authors: Paradis Pierre , Mahmoud Ahmad , Caillon Antoine , Shokoples Brandon , Ferreira Nathanne , Comeau Kevin , Hatano Shinya , Yoshikai Yasunobu , Lewis Julia , Tigelaar Robert , Schiffrin Ernesto
Published: October 2024

Background:
γδ T cells mediate angiotensin II (AngII)-induced blood pressure (BP) elevation, vascular injury and activated T cell infiltration in mesenteric vessels with adherent perivascular adipose tissue (MV/PVAT). γδ T cells expressing specific T cell receptor (TCR) variable (V) chain γ develop in several waves in the thymus and migrate to diverse tissues. We hypothesized that γδ T cells expressing specific Vγ subtypes in perivascular tissue are implicated in AngII detrimental hypertensive effects.

Methods:
C57BL/6J male mice were infused or not with AngII (490 ng/kg/min, SC) for 14 days. Vγ1 and Vγ2 (Vγ1/2)
+
, Vγ4
+
, Vγ5
+
, Vγ6
+
, and Vγ7
+
γδ T cell frequencies and T cell phenotypes were determined by flow cytometry in the spleen, descending thoracic aorta (DTAo)/PVAT, and MV/PVAT (n=7-13). Other sets of AngII-infused mice were injected with control or anti-Vγ6 (n=7-12) or Vγ4 antibodies (n=5-8). BP was determined by telemetry, mesenteric artery (MA) function and remodeling by pressurized myography, and T cell phenotypes by flow cytometry.

Results:
In DTAo/PVAT and MV/PVAT, respectively, 43±5% and 51%±6% of γδ T cells were Vγ6
+
, and 17±2% and 23±4% Vγ4
+
, whereas in the spleen, 36±2% of the γδ T cells were Vγ1/2
+
, 32±1% Vγ4
+
and 24±1% Vγ6
+
. AngII increased the frequency of Vγ6
+
γδ T cells in the spleen (0.8±0.1 vs 0.5±0.0% of CD3,
P
<0.001) and DTAo/PVAT (5.9±0.8 vs 3.6±0.5% of CD3,
P
<0.05), and tended to increase their frequency in MV/PVAT. The majority of Vγ6
+
γδ T cells were activated (CD69
+
) in DTAo/PVAT (75±5%) and MV/PVAT (84±2%). Vγ6
+
γδ T cell depletion caused a steeper BP elevation (
P
<0.05) and worsened MA endothelial dysfunction (maximal acetylcholine relaxation response: 28±3 vs 55±8%,
P
<0.05) in mice infused with AngII. This was associated with increased CD69
+
non-Vγ6
+
γδ T cells in DTAo/PVAT (3.0±0.7 vs 0.8±0.4% of CD3,
P
<0.01) and MV/PVAT (15.2±2.6 vs 4.9±1.0% of CD3,
P
<0.01). Depletion of Vγ4
+
γδ T cells in perivascular tissues, did not alter the AngII detrimental effects.

Conclusions:
Vγ6
+
γδ T cells are the most abundant γδ T cell Vγ subtype in perivascular tissues. AngII increased Vγ6
+
γδ T cells in the spleen and perivascular tissue. Vγ6
+
γδ T cell depletion in AngII-infused mice induced a steeper BP elevation, worsened MA endothelial dysfunction, and activated non-Vγ6
+
γδ T cells. Vg6
+
γδ T cells may play a protective role in AngII-induced hypertension and vascular injury.
Abstract 36: Ablation of T-Lymphocyte β-Arrestins Reduces Inflammation and Increases Salt Retention in the Absence of Blood Pressure Changes Following Angiotensin II Infusion
Authors: Ghobrial Mina , Fekete Eva , Muskus Veitia Patricia , Brozoski Daniel , Lu Ko-Ting , Hantke Guixa Ana , Wackman Kelsey , Sigmund Curt , Nakagawa Pablo
Published: October 2024

Introduction:
T-cells contribute to hypertension (HT) development. Among several G-protein coupled receptors regulating T-cell function, it has been hypothesized that some subpopulations use the angiotensin II (ANG II) type 1 receptor (AT
1
R) to modulate immune responses in HT. Selective ablation of AT
1
Rs in T-cells paradoxically increases hypertensive end-organ damage (EOD). While AT
1
Rs typically induces classical G-protein signaling, recent studies have uncovered a protective signaling cascade
via
β-arrestin (
ARRB
) proteins. We hypothesize that T-cell
ARRB
s mediate an anti-hypertensive and anti-inflammatory role in response to ANG II, decreasing EOD in HT.

Methods:
Breeding CD4-Cre mice with
Arrb1
-Flox or
Arrb2
-Flox mice generated lines with T-cell-specific deficiency of
Arrb1
or
Arrb2
genes. Hypertensive conditions modeling kidney damage consisted of unilateral nephrectomy, 4% salt diet, and continuous infusion of 1 mg/kg/min ANG II (UNX/HSD/ANG). Age/sex-matched Cre
-
littermates were used as controls (WT). Blood pressure (BP) was measured at baseline and for the following three weeks using radiotelemetry.

Results:
At baseline, there were no significant differences in systolic BP between groups (WT:124.1±8.0 mmHg; CD4
Arrb1-null
:125.4±8.1; CD4
Arrb2-null
:128.7±3.1;
p
=0.37; n=12/10/8). Three weeks of UNX/HSD/ANG equally increased systolic BP in all groups (WT:178.0±14.0 mmHg; CD4
Arrb1-null
:174.8±26.3; CD4
Arrb2-null
:179.9±11.1;
p
=0.86; n=7/9/7). Despite similar BPs, CD4
Arrb2-null
mice demonstrated less renal inflammation than WT after four weeks of UNX/HSD/ANG, with a reduction in interleukin-1α protein (3.8±3.2
vs.
8.3±2.8
p
<0.05; n=4/6) and a trend towards decreased tumor necrosis factor-α transcript (0.63-fold change
vs.
1.9,
p
=0.051; n=10/8). Furthermore, CD4
Arrb1-null
mice increased 24-hour voluntary water intake compared to WT (15.7±3.4 mL
vs.
12.2±1.6;
p
<0.05; n=12/7), and 24-hour urine output (10.0±3.7 mL/d
vs.
6.8±1.5;
p
<0.05; n=11/8). However, there was no difference in sodium excretion during this period (1.9±0.5 eq/d
vs.
1.7±0.1;
p
=0.14; n=11/7). Interestingly, 4 hours after intraperitoneal saline injection, CD4
Arrb1-null
mice excreted significantly less sodium compared to WT (5.6±3.5%
vs
. 31.4±10.6%;
p
<0.05; n=4/5).

Conclusion:
Contrary to our hypothesis, these data suggest that T-cell
Arrb
s do not protect against the hypertensive response to ANG II, but instead play a complicated role in the modulation of hydromineral balance and inflammation.
Abstract 37: Targeted Deletion of NKCC1 in Vascular Smooth Muscle Cells Lowers Blood Pressure in Normotensive and Hypertensive Mice
Authors: Seniuk Anika , Steffen Claudius , Vitzthum Helga , Hubner Christian , Ehmke Heimo
Published: October 2024

Background:
Salt plays a major role in blood pressure homeostasis and the pathogenesis of arterial hypertension. While numerous mechanisms have been described by which Na
+
can influence arterial blood pressure, the role of Cl
-
in blood pressure regulation is still largely unexplored. The Na
+
-K
+
-2Cl
-
cotransporter NKCC1 is an important Cl
-
import pathway in vascular smooth muscle cells (VSMC). To investigate the role of NKCC1 in blood pressure regulation, we generated mice with an inducible VSMC-specific disruption of
Slc12a2
.

Methods:
Mean arterial blood pressure and heart rate were measured by radiotelemetry in unrestrained mice lacking vascular NKCC1 (KO) and wildtype littermates (WT). Cardiac function was assessed by echocardiography. Plasma renin activity and aldosterone concentrations were quantified by radioimmunoassay. Protein expression of renal sodium transporters and channels was analyzed by Western blotting.

Results:
Mean arterial blood pressure was markedly decreased in
Nkcc1
-deficient mice 6 weeks after induction of gene disruption (from 116±2 mm Hg to 103±4 mm Hg, n=18, p<0.001, 2-way ANOVA and Bonferroni post-test), while it did not change in WT control animals (n=17). Heart rate, stroke volume, fractional shortening, and ejection fraction remained unchanged compared to non-induced controls. We did not observe a compensatory increase in plasma renin activity or aldosterone levels, and the blood pressure response to ANG II receptor 1 antagonism by losartan treatment for 7 days was similar in both groups (ΔMAP KO -8.8±3.0 mm Hg, n=8, ΔMAP WT -12.4±1.9 mm Hg, n=7; p>0.05, 2-way ANOVA). In the kidney, the relative expression of total NCC (1.5-
fold
, n=7-8; p=<0.05, Student’s t-test) and pNCCThr53 (1.9-
fold
, n=7-8; p=<0.05, Student’s t-test) were modestly increased. The hypotensive effect of
Nkcc1
disruption was maintained after induction of hypertension by either infusion of ANG II for 10 days (ΔMAP KO 43.8±6.5 mm Hg, n=6, ΔMAP WT 50.2±7.2 mm Hg, n=4; p>0.05, 2-way ANOVA) or a high salt diet for 10 days (ΔMAP KO 21.7±7.0 mm Hg, n=8, ΔMAP WT 34.0±7.3 mm Hg, n=5; p>0.05, 2-way ANOVA).

Conclusions:
Our findings indicate that NKCC1 in vascular smooth muscle contributes significantly to the maintenance of arterial blood pressure. Inhibition of NKCC1 in VSMC may provide additional blood pressure reduction in resistant hypertension.
Abstract 40: Sodium-mediated Blood Pressure Increase in Chronic Kidney Disease and Diabetic Kidney Disease Patients Coincides With Vasodysfunction.
Authors: van Duin Robert , Oppelaar Jetta , Van den Born Bert-Jan , Olde Engberink Rik , Vogt Liffert
Published: October 2024

Background:
Chronic kidney disease (CKD) and diabetic kidney disease (DKD) patients are advised to limit salt intake, because of its detrimental effects on blood pressure (BP) and albuminuria. The salt-sensitive BP response in these patients may be attributed to impaired renal ability to excrete sodium (Na+) resulting in fluid overload and/or vasodysfunction. To define which factor relates to BP sensitivity in both CKD and DKD, we studied the Na+-mediated BP response in these patients as compared to healthy volunteers (HV).

Methods:
In this randomized crossover study, CKD patients (KDIGO stage G2/3A, proteinuria >0.5 g/d), Type 2 DKD patients (KDIGO stage G2/3A, proteinuria 0.2 - 0.5 g/d) and HV followed a 7-day low sodium diet (LSD, <50mmol/d) and high sodium diet (HSD, >200mmol/d), respectively. After each diet, assessment of the hemodynamic profile included daytime BP (Mobil-O-Graph) and cardiac output (CO) measurements (using NexfinTM), by which SVR was calculated accordingly. Extracellular fluid volume (ECV) change was assessed with multi-frequency body impedance spectroscopy (Fresenius).

Results:
We included 20 CKD (mean age 48 yrs, median (IQR) proteinuria 0.7 (0.6-1.6) g/d, mean (SD) eGFR 60.7 (16.3) ml/min/1.73m2) and 8 DKD patients (age 68 yrs, proteinuria 0.3 (0.2-0.4) g/d, eGFR 59.0 (15.9) ml/min/1.73m2), and 16 HV (age 41 yrs). Dietary salt intervention was successful with mean difference in urine Na+ excretion of 185, 168, and 209 mmol/day in CKD, DKD and HV respectively. After HSD, mean (SD) systolic BP change was 9.3 mmHg (8.9; p<0.001) in CKD and 13.8 mmHg (10.4; p<0.01) in DKD, while systolic BP change (3.1 mmHg (6)) in HV was not significant. After HSD, we observed significant ECV expansion in all groups without differences between groups (CKD 1.6 L, DKD 1.6 L, HV 1.4 L, p=0.27), while no effects on CO were observed. After HSD, the mean (SD) SVR in CKD increased with 125.7 dyn*sec*cm-5 (247.9, p=0.04) and in DKD with 125.5 dyn*sec*cm-5 (142.6, p=0.04), while in HC a non-significant decline of -109.2 dyn*sec*cm-5 was observed (358.2, p=0.24).

Conclusion:
The Na+-mediated BP rise in both CKD and DKD coincides with an increase in SVR. The absence of differences in ECV and CO between the three groups indicates that salt sensitivity in both CKD and DKD is not completely based on fluid overload but might be due to an inability to induce vasodilation.
Abstract 41: SerpinA3N in Leptin Receptor Neurons Regulates Metabolic Homeostasis and Cardiovascular Function
Authors: Guo Deng-Fu , Morgan Donald , Olson Alexis , Rahmouni Kamal
Published: October 2024

Obesity and associated conditions such as type 2 diabetes are among the most concerning health issues in the U.S. Leptin receptor (LepR) signaling in the brain plays a pivotal role in the regulation of food intake and energy expenditure. SerpinA3N is a serine protease inhibitor which is highly expressed in the arcuate nucleus of the hypothalamus, upregulated in diet-induced obesity and by leptin stimulation. However, the role of SerpinA3N in the LepR expressing cells in the control of body weight and glucose homeostasis remain unknown. RNAseq analysis confirmed that SerpinA3N mRNA levels were significantly upregulated in the hypothalamus of mice fed high fat high sucrose diet (HFHSD) for 12 weeks. Using immunofluorescence staining, we confirmed that SerpinA3N is expressed the LepR positive cells of the hypothalamus. Next, we investigated whether loss of SerpinA3N in LepR expressing neurons affects metabolic and glucose homeostasis. For this, we crossed mice harboring floxed alleles of the
SerpinA3N
gene (SerpinA3N
fl/fl
) with mice expressing Cre recombinase driven by the LepR (LepRb
Cre
). We found that female, but not male, conditional knockout (CKO, LepRb
Cre
/SerpinA3N
fl/fl
) mice have significantly lower body weight when fed HFHSD. This lower body weight was due to decreased fat mass, measured by NMR. However, no alteration in food intake was observed between female CKO mice and controls, indicating that increased energy expenditure may account for the lower body weight in female CKO mice. Interestingly, CKO mice displayed exaggerated weight loss and anorectic response to leptin treatment (1 mg/g, twice daily, IP), indicating that loss of SerpinA3N enhances leptin sensitivity. Female CKO mice fed normal chow diet also exhibited enhanced insulin sensitivity compared to control animals. On the other hand, male CKO mice displayed significantly improved glucose handling, but no change in insulin sensitivity. Insulin-induced phosphorylation of AKT was elevated only in skeletal muscleof CKO mice relative to controls. Finally, systolic arterial blood pressure of CKO mice fed HFHSD trend to be lower during dark time compared to controls (142±6.4 vs 127.9±7.1 mmHg, p=0.06), although heart rate was not changed. These results demonstrate that SerpinA3N in LepRb-expressing neurons regulate energy and glucose homeostasis in a sex-dependent manner,and may protect from HFHSD induced hypertension.
Abstract 46: Reduced Angiotensin II Type 2 Receptor-Mediated Responses Contribute to Increased Angiotensin II Vasoconstrictor Sensitivity in Otherwise Healthy Women With a History Of Preeclampsia
Authors: Schwartz Kelsey , Jalal Diana , Stanhewicz Anna
Published: October 2024

Women with a history of preeclampsia (hxPE) have a ≥4-fold risk for developing cardiovascular disease compared to matched women who had a healthy pregnancy (hxHC). Angiotensin (ang) II-mediated vasoconstriction contributes to the persistently reduced microvascular function in the years after preeclampsia. Excessive activation of ang II type 1 receptors (AT
1
R) is one putative mechanism underlying increased ang II constrictor sensitivity. This may be countered by ang II type 2 receptor (AT
2
R) activation; however, the extent that reductions in AT
2
R-mediated responses contribute to increased ang II sensitivity after preeclampsia is unknown. Using the cutaneous microcirculation as a model, we hypothesized that 1) AT
2
R-mediated dilation is attenuated in hxPE compared with hxHC, 2) AT
1
R inhibition improves reduced AT
2
R-mediated dilation in hxPE, and 3) AT
2
R inhibition would increase ang II-mediated constriction in hxHC but have no effect in hxPE. Nine hxPE (37±5 years) and 9 matched hxHC (34±5 years) within 5 years postpartum had 4 intradermal microdialysis fibers placed in the ventral forearm for the local delivery of pharmacological agents. Two fibers received graded infusions of compound 21 (AT
2
R agonist; 10
-14
-10
-8
mol/L) alone or co-perfused with losartan (AT
1
R antagonist; 43µmol/L) for the assessment of AT
2
R-mediated dilation. The remaining 2 sites received graded infusions of ang II (10
−20
–10
−4
mol/L) alone or co-perfused with PD-123319 (AT
2
R antagonist; 1µmol/L) to assess the role of AT
2
R in vasoconstrictor sensitivity to ang II. Red blood cell flux was measured directly over each fiber and cutaneous vascular conductance was calculated (CVC=flux/MAP) and expressed as a % maximum (43°C + 28mM SNP) or % baseline. Those with hxPE had attenuated AT
2
R-mediated dilation (hxPE: 15±5 vs hxHC: 27±9%max; P=0.01) that was improved with AT
1
R inhibition (losartan: 22±7 vs control: 15±5%max; P=0.003). Vasoconstrictor sensitivity to ang II was greater in hxPE compared with hxHC (hxPE: 39±14 vs hxHC: 55±6%baseline; P<0.001). AT
2
R inhibition increased the vasoconstrictor response to ang II in hxHC (PD-123319: 39±12 vs control: 55±65%baseline; P<0.001) but had no effect in hxPE (PD-123319: 39±13 vs control: 39±14%baseline; P=0.29). These data suggest that women with a history of preeclampsia have reductions in AT
2
R-mediated dilation that contributes to increased ang II vasoconstrictor sensitivity and sustained microvascular dysfunction after preeclampsia.
Abstract 50: The Comparison of Office Blood Pressure with Morning Home Blood Pressure: Self-Automated Measurement
Authors: Suntornlekha Nutnicha , Kuanprasert Srun , Chai-Adisaksopha Chatree , Jesadaporn Panas , Mekara Warangkana , LEEMASAWAT KRIT
Published: October 2024

Background:
Morning hypertension is associated with poor cardiovascular outcomes. However, office blood pressure (OBP) remains a gold standard in hypertension monitoring and management. Unfortunately, the accuracy in OBP measurement is problematic.

Objectives:
This study aimed to determine whether routine or unattended OBP could accurately represent home blood pressure (HBP) in patients with hypertension.

Methods:
We conducted a single-center, cross-sectional study. Hypertensive patients aged between 18 and 80 years, capable of performing self-automated blood pressure measurement, were included in the study. Three self-automated methods of blood pressure measurement were performed: Routine OBP, unattended OBP, and HBP. All participants were instructed to rest for at least 5 minutes prior to measuring blood pressure, and HBP was recorded twice daily, in the morning and at night, for a week after the enrolled visit without antihypertensive medication adjustment.

Results:
Overall 190 consecutive patients were enrolled, with a mean age of 65.2 ±7.5 years old. Half of the patients had coronary artery disease or type 2 diabetes as comorbidities. Routine OBP was significantly higher than unattended OBP and HBP. Interestingly, Systolic values of unattended OBP were similar to those of morning HBP (mean difference -0.6 ±16.0 mmHg, p=0.641) with moderate correlation (r=0.516, p<0.001). Although diastolic values of unattended OBP were lower than those of morning HBP (mean difference -3.3 ±9.0 mmHg, p<0.001), the values were moderately correlated (r=0.581, p<0.001). Bland-Alman plots between unattended OBP and morning HBP were shown in Figure 1. Both systolic and diastolic values of unattended OBP and average HBP were significantly different (mean difference -3.5 ±14.9 mmHg, p=0.002, and 1.6 ±8.6 mmHg, p=0.013, respectively).

Conclusion:
Unattended OBP method could effectively represent morning HBP, thus advocating to use in clinical practice.