bluuebunny/crossref_metadata_2025 huggingface dataset:
Assessment of Micronuclei Frequency in the Peripheral Blood of Adult and Pediatric Patients Receiving Fractionated Total Body Irradiation
Authors: Kanagaraj Karthik , Phillippi Michelle A. , Narayan Pratyush , Szolc Barbara , Perrier Jay R. , McLane Amanda , Wolden Suzanne L. , Barker Christopher A. , Wang Qi , Amundson Sally A. , Brenner David J. , Turner Helen C.
Published: October 2023

The cytokinesis-block micronucleus (CBMN) assay is an established method for assessing chromosome damage in human peripheral blood lymphocytes resulting from exposure to genotoxic agents such as ionizing radiation. The objective of this study was to measure cytogenetic DNA damage and hematology parameters in vivo based on MN frequency in peripheral blood lymphocytes (PBLs) from adult and pediatric leukemia patients undergoing hematopoietic stem cell transplantation preceded by total body irradiation (TBI) as part of the conditioning regimen. CBMN assay cultures were prepared from fresh blood samples collected before and at 4 and 24 h after the start of TBI, corresponding to doses of 1.25 Gy and 3.75 Gy, respectively. For both age groups, there was a significant increase in MN yields with increasing dose (p < 0.05) and dose-dependent decrease in the nuclear division index (NDI; p < 0.0001). In the pre-radiotherapy samples, there was a significantly higher NDI measured in the pediatric cohort compared to the adult due to an increase in the percentage of tri- and quadri-nucleated cells scored. Complete blood counts with differential recorded before and after TBI at the 24-h time point showed a rapid increase in neutrophil (p = 0.0001) and decrease in lymphocyte (p = 0.0006) counts, resulting in a highly elevated neutrophil-to-lymphocyte ratio (NLR) of 14.45 ± 1.85 after 3.75 Gy TBI (pre-exposure = 4.62 ± 0.49), indicating a strong systemic inflammatory response. Correlation of the hematological cell subset counts with cytogenetic damage, indicated that only the lymphocyte subset survival fraction (after TBI compared with before TBI) showed a negative correlation with increasing MN frequency from 0 to 1.25 Gy (r = −0.931; p = 0.007). Further, the data presented here indicate that the combination of CBMN assay endpoints (MN frequency and NDI values) and hematology parameters could be used to assess cytogenetic damage and early hematopoietic injury in the peripheral blood of leukemia patients, 24 h after TBI exposure.
Strategies to Mitigate the Drug-Drug Interaction between Nirmatrelvir/Ritonavir and Tacrolimus in Allogeneic Hematopoietic Stem Cell Transplant Recipients on Azole Antifungals: Results of a Case Series
Authors: Griffin Shawn P. , Lee Benjamin , Doh Jean , Paradyse Alexander R. , Jeyakumar Deepa , Arter Zhaohui , Nam Hannah , Blodget Emily , Smith Julie , Valek Anitram , Vittayawacharin Pongthep , Kongtim Piyanuch , Ciurea Stefan O.
Published: October 2023

Introduction: Nirmatrelvir/ritonavir (NIM/r) inhibits tacrolimus metabolism resulting in a profound drug-drug interaction that is further complicated by the use of azole antifungals. Case Presentations: We describe three strategies, in 4 patient cases, for the initiation of NIM/r in allogeneic hematopoietic stem cell transplant (alloHSCT) recipients on tacrolimus at the time of diagnosis. Patients 1 and 2 (strategy 1) experienced prolonged, elevated tacrolimus concentrations after an empiric 33% reduction in tacrolimus dose and adjustment of azole antifungal at NIM/r initiation (strategy 1) and with complete discontinuation of tacrolimus and azole antifungal at NIM/r initiation (strategy 2). Patients 3 and 4 (strategy 3) did not experience elevated tacrolimus concentrations on NIM/r treatment with complete discontinuation of tacrolimus and azole antifungal and a 12–24-h delay in NIM/r initiation. Reinitiation of tacrolimus after NIM/r completion resulted in variable tacrolimus concentrations. Conclusion: NIM/r-tacrolimus is a serious drug-drug interaction which can be mitigated by early discontinuation of tacrolimus and azole antifungals, close monitoring, and reinitiation of tacrolimus and antifungal 48–72 h after completion of therapy.
Regional Variation in the Use of Percutaneous Kidney Biopsy in Japan
Authors: Oda Yasuhiro , Nishi Hiroshi , Nangaku Masaomi
Published: October 2023

Introduction: The regional variation in the use of percutaneous kidney biopsy in Japan remains unknown. There are several large datasets of kidney biopsies in Japan, but an exhaustive survey of kidney biopsies is lacking. Methods: We analyzed insurance claims for percutaneous kidney biopsies registered in the National Database of Health Insurance Claims and Specific Health Checkups of Japan, which is the closest to a complete dataset of kidney biopsies performed in Japan. In combination with other nationwide survey results, the number of inpatient percutaneous kidney biopsies per population in each prefecture was calculated. Factors associated with the frequency of percutaneous kidney biopsies were also explored. Results: The database contained 22,419 health insurance claims for percutaneous kidney biopsy in the fiscal year 2020. The frequency of inpatient percutaneous kidney biopsies could be up to 4.8 times as frequent in one prefecture than in another, even after adjusting for age and sex. The frequency of inpatient percutaneous kidney biopsies showed a positive correlation with the number of annual kidney transplants and patients on peritoneal dialysis per population and a weak negative correlation with the prevalence of reduced kidney function in the population aged 40–74 years. Conclusion: We found a large regional variation in the frequency of inpatient percutaneous kidney biopsies. Kidney transplants and peritoneal dialysis might be offered more frequently in regions with a higher frequency of kidney biopsy. This is the first dataset that shows more than 20,000 kidney biopsies were performed per year in Japan, as of 2020.
Association between Low Vitamin D Status, Serotonin, and Clinico-Biobehavioral Parameters in Alzheimer’s Disease
Authors: Richter Anna-Lena , Diepeveen-de Bruin Marlies , Balvers Michiel G.J. , De Groot Lisette C.P.G.M. , De Deyn Peter Paul , Engelborghs Sebastiaan , Witkamp Renger F. , Vermeiren Yannick
Published: October 2023

Introduction: Studies suggest a role of vitamin D in the progression and symptomatology of Alzheimer’s disease (AD), with few in vitro studies pointing to effects on serotonergic and amyloidogenic turnover. However, limited data exist in AD patients on the potential association with cognition and behavioral and psychological signs and symptoms of dementia (BPSD). In this retrospective cross-sectional study, we, therefore, explored potential correlations of serum 25-hydroxyvitamin D3 (25(OH)D3) concentrations, indicative of vitamin D status, with serum serotonin (5-hydroxytryptamine, 5-HT) levels, cognitive/BPSD scorings, and cerebrospinal fluid (CSF) biomarker levels. Methods: Frozen serum samples of 25 well-characterized AD subjects as part of a previous BPSD cohort were analyzed, of which 15 had a neuropathologically confirmed diagnosis. Serum 25(OH)D3 levels were analyzed by means of LC-MS/MS, whereas 5-HT concentrations were quantified by competitive ELISA. Results: Among AD patients, vitamin D deficiency was highly prevalent, defined as levels below 50 nmol/L. Regression analyses, adjusted for age, gender, and psychotropic medications, revealed that serum 25(OH)D3 and 5-HT levels were positively associated (p = 0.012). Furthermore, serum 25(OH)D3 concentrations correlated inversely with CSF amyloid-beta (Aβ1-42) levels (p = 0.006), and serum 5-HT levels correlated positively with aggressiveness (p = 0.001), frontal behavior (p = 0.001), depression (p = 0.004), and partly with cognitive performance (p < 0.005). Lastly, AD patients on cholinesterase inhibitors had higher serum 25(OH)D3 (p = 0.030) and lower serum 5-HT (p = 0.012) levels. Conclusions: The molecular associations between low vitamin D status, serum 5-HT, and CSF Aβ1-42 levels are highly remarkable, warranting further mechanistic and intervention studies to disclose potential involvement in the clinico-biobehavioral pathophysiology of AD.
Admission Hypocalcemia and the Need for Endoscopic and Clinical Interventions among Patients with Upper Gastrointestinal Bleeding
Authors: Korytny Alex , Mazzawi Fares , Marcusohn Erez , Klein Amir , Epstein Danny
Published: October 2023

Introduction: Calcium is an essential co-factor in the coagulation cascade, and hypocalcemia is associated with adverse outcomes in bleeding patients, including trauma patients, women with postpartum hemorrhage, and patients with intracranial hemorrhage. In this retrospective, single-center, cohort study, we aimed to determine whether admission-ionized calcium (Ca++) is associated with higher rates of therapeutic interventions among patients presenting with acute nonvariceal upper gastrointestinal bleeding (NV-UGIB). Methods: Adult patients admitted due to NV-UGIB between January 2009 and April 2020 were identified. The primary outcome was defined as a need for clinical intervention (two or more packed cell transfusions, need for endoscopic, surgical, or angiographic intervention). Univariate and multivariable logistic regression analyses were performed to determine whether Ca++ was an independent predictor of the need for therapeutic interventions. Propensity score matching was performed to adjust the imbalances of covariates between the groups. Results: A total of 434 patients were included, of whom 148 (34.1%) had hypocalcemia (Ca++ <1.15 mmol/L). Patients with hypocalcemia were more likely to require therapeutic interventions than those without hypocalcemia (48.0% vs. 18.5%, p < 0.001). Specifically, patients with hypocalcemia were more likely to require endoscopic intervention for control of bleeding (25.0% vs. 15.7%, p = 0.03) and multiple packed cell transfusions (6.8% vs. 0.3%, p < 0.001). Additionally, they had significantly longer hospital stay (5.0 days [IQR 3.0–8.0] vs. 4.0 days [IQR 3.0–6.0], p = 0.01). After adjusting for multiple covariates, Ca++ was an independent predictor of the need for therapeutic intervention (aOR 1.62, 95% confidence interval [CI] 1.22–2.14, p < 0.001). The addition of Ca++ to the Modified Glasgow Blatchford score improved its accuracy in the prediction of therapeutic intervention from AUC of 0.68 (95% CI 0.63–0.72) to 0.72 (95% CI 0.67–0.76), p = 0.02. After incorporation of the propensity score, the results did not change significantly. Conclusion: These findings suggest that hypocalcemia is common and is associated with an adverse clinical course in patients with NV-UGIB. Measurement of Ca++ on admission may facilitate risk stratification in these patients. Trials are needed to assess whether the correction of hypocalcemia will lead to improved outcomes.
COVID-19 in Patients with Chronic Lymphocytic Leukemia: What Have We Learned?
Authors: Harel Reut , Itchaki Gilad
Published: October 2023

Background: Chronic lymphocytic leukemia (CLL) is a prevalent hematological malignancy (HM) characterized by inherent immunodeficiency, which is further pronounced by disease-directed therapy. The COVID-19 pandemic has had devastating outcomes, and although its impact has diminished over time, it continues to be a cause of significant morbidity and mortality, particularly among immunodeficient patients. Summary: In this review, we describe mechanisms of immune dysfunction in CLL in relation to COVID-19, provide an overview of the clinical outcomes of the disease in this patient population, and identify risk factors associated with severe morbidity and mortality. Additionally, we acknowledge the influence of the rapidly evolving landscape of new disease variants. The review further delineates the humoral and cellular responses to vaccination and their clinical efficacy in preventing COVID-19 in CLL patients. Moreover, we explore potential approaches to enhance these immune responses. Pre- and post-exposure prophylaxis strategies are discussed, along with description of common agents in the treatment of the disease in both outpatient and inpatient setting. Throughout the review, we emphasize the interplay between novel therapies for CLL and COVID-19 outcomes, prevention, and treatment and describe the impact of COVID-19 on the utilization of these novel agents. This information has the potential to guide clinical decision making in the management CLL patients. Key Messages: CLL patients are at risk for severe COVID-19 infection. Vaccinations and COVID-19 directed therapy have improved outcomes in patients with CLL, yet clinical challenges persist.
ОСВІТНЬО-НАУКОВА ПРОГРАМА ЗА СПЕЦІАЛЬНІСТЮ 224 «ТЕХНОЛОГІЇ МЕДИЧНОЇ ДІАГНОСТИКИ ТА ЛІКУВАННЯ»
Authors: Мінцер О.П. , Бабкіна Т. М. , Щербіна О. В. , Клименко С. В. , Краснов В. В. , Король П. О. , Сергієнко Л. І.
Published: October 2023

Розвиток галузі охорони здоров'я неможливий без застосування сучасних технологій, що останнім часом стають одним із вирішальних факторів для забезпечення якісної та ефективної медичної діагностики та лікування пацієнтів. Завдяки залученню інформаційно-комунікаційних і цифрових технологій, мі-кроелектроніки виникли такі напрями як телемедицина, цифрова охорона здоров'я, мобільна медицина, 3D-друк (адитивні технології) та інші. В той же час без лабораторних та інструментальних досліджень неможливою стає не тільки постановка клінічного діагнозу, але й контроль над безпекою лікування. Від рівня достовірності досліджень залежить прийняття лікарем рішень про діагностування, профілактику та лікування пацієнта. Представлено сучасне уявлення про програму навчання майбутніх спеціалістів ступеня доктора філософії за спеціальністю 224 «Технології медичної діагностики та лікування» галузі знань 22 «Охорона здоров'я». Методами експертного оцінювання проаналізовано розвиток теорії і практики застосування технологій діагностики та лікування захворювань внутрішніх органів, у тому числі інформаційних. Уніфіковано сучасні технології клінічних лабораторних і променевих методів дослідження, що сприяють підвищенню якості надання медичної допомоги; методи променевої терапії, що сприяють підвищенню ефективності лікування онкологічних і неонкологічних захворювань, а також методи прийняття рішень у діагностиці, лікуванні та прогнозуванні станів пацієнта.
По завершенню навчання, здобувачі вищої освіти третього рівня мають оволодіти загальнонаукови-ми компетентностями, здобути універсальні навики дослідника та мовні компетентності. Тематика курсів професійної підготовки розроблена з метою сприяння здатності інтегрувати знання та розв'язувати складні завдання в мультидисциплінарних і трансдисциплінарних контекстах. Отже, зміст запропонованої освітньо-науково'! програми за спеціальністю 224 «Технології медичної діагностики та лікування» направлено на підготовку висококваліфікованих і конкурентоспроможних спеціалістів ступеня доктора філософії та здобуття ними в процесі навчання необхідних теоретичних знань, що носять мультидисциплінарний характер, та комплексу практичних умінь, навиків і компетентностей для виконання інноваційних наукових досліджень та впровадження отриманих результатів у практичну охорону здоров'я.
MRP1-Dependent Extracellular Release of Glutathione Induces Cardiomyocyte Ferroptosis After Ischemia-Reperfusion
Authors: Ichihara Genki , Katsumata Yoshinori , Sugiura Yuki , Matsuoka Yuta , Maeda Rae , Endo Jin , Anzai Atsushi , Shirakawa Kohsuke , Moriyama Hidenori , Kitakata Hiroki , Hiraide Takahiro , Goto Shinichi , Ko Seien , Iwasawa Yuji , Sugai Kazuhisa , Daigo Kyohei , Goto Shinya , Sato Kazuki , Yamada Ken-ichi , Suematsu Makoto , Ieda Masaki , Sano Motoaki
Published: October 2023

BACKGROUND:

The membrane components of cardiomyocytes are rich in polyunsaturated fatty acids, which are easily oxidized. Thus, an efficient glutathione-based lipid redox system is essential for maintaining cellular functions. However, the relationship between disruption of the redox system during ischemia-reperfusion (IR), oxidized lipid production, and consequent cell death (ferroptosis) remains unclear. We investigated the mechanisms underlying the disruption of the glutathione-mediated reduction system related to ferroptosis during IR and developed intervention strategies to suppress ferroptosis.

METHODS:

In vivo fluctuations of both intra- and extracellular metabolite levels during IR were explored via microdialysis and tissue metabolome analysis. Oxidized phosphatidylcholines were assessed using liquid chromatography high-resolution mass spectrometry. The areas at risk following IR were assessed using triphenyl-tetrazolium chloride/Evans blue stain.

RESULTS:

Metabolomic analysis combined with microdialysis revealed a significant release of glutathione from the ischemic region into extracellular spaces during ischemia and after reperfusion. The release of glutathione into extracellular spaces and a concomitant decrease in intracellular glutathione concentrations were also observed during anoxia-reperfusion in an in vitro cardiomyocyte model. This extracellular glutathione release was prevented by chemical inhibition or genetic suppression of glutathione transporters, mainly MRP1 (multidrug resistance protein 1). Treatment with MRP1 inhibitor reduced the intracellular reactive oxygen species levels and lipid peroxidation, thereby inhibiting cell death.

Subsequent in vivo evaluation of endogenously oxidized phospholipids following IR demonstrated the involvement of ferroptosis, as levels of multiple oxidized phosphatidylcholines were significantly elevated in the ischemic region 12 hours after reperfusion. Inhibition of the MRP1 transporter also alleviated intracellular glutathione depletion in vivo and significantly reduced the generation of oxidized phosphatidylcholines. Administration of MRP1 inhibitors significantly attenuated infarct size after IR injury.

CONCLUSIONS:

Glutathione was released continuously during IR, primarily in an MRP1-dependent manner, and induced ferroptosis. Suppression of glutathione release attenuated ferroptosis and reduced myocardial infarct size following IR.
Abstract 017: Differential Regulation Of Piezo1 And Major Histocompatibility Complex I And II In Male And Female Renal Endothelial Cells During Ang II-mediated Hypertension
Authors: Colvert Alex , McCrorey Marice , Hawkins Kennedy , Brooks Heddwen L , Deleon-Pennell Kristine Y , Van Beusecum Justin P
Published: October 2023

Infiltration of immune cells to the kidneys contributes to development of hypertension (HTN) and renal injury. One facet in immune cell diapedesis and transmigration is endothelial activation, possibly through a mechanosensitive ion channel, Piezo1, which is expressed on endothelial and immune cells. It has been demonstrated that both male and female renal endothelial cells possess the ability to regulate major histocompatibility complex (MHC) I and II in response to HTN-induced biomechanical stimuli. The
in vivo
mechanisms by which endothelial MHCs are regulated in response to HTN remains unknown. We hypothesized that HTN-induced mechanical stress on the renal endothelium stimulates upregulation of MHCs in a Piezo1-dependent manner. Male and female C57Bl/6J mice 10 to 12 weeks of age (n=4-5/group) were treated with angiotensin II (ang II; 490ng/kg/min) or vehicle by subcutaneous osmotic minipump for 28-days. Then kidneys were excised for immunofluorescence staining of MHC I, II and Piezo1. Primary mouse renal endothelial cells (MRECs) were isolated via magnetic sorting using CD31
+
microbeads to measure
H2-db
and
H2-ab
and
Piezo1
by real-time PCR (RT-PCR) and immunoblotting. Immunofluorescence microscopy demonstrated that MHC I, MHC II, and Piezo1 were present in the renal endothelium of male and females irrespective of treatment. Ang II significantly reduced
H2-ab
expression (p<0.001) in endothelial cells in male mice, with no significant difference in
H2-db
expression. Additionally, there were trends for downregulation and upregulation of
Piezo1
, (p=0.11) and
Vcam1
(p=0.15), respectively. Ang II infusion in males showed significant reduction in protein expression of renal endothelial Piezo1 (p<0.05), a trend for increased expression of MHC II, and no change in MHC I. In comparison, Ang II induced no significant differences in mRNA expression of
Piezo1
,
Vcam1
,
H2-Db
, or
H2-Ab
in isolated endothelial cells, or in the protein expression of Piezo1, MHC I and MHC II in female ang II treated mice. Our data identify a sex difference in the regulation of endothelial Piezo1 in the kidney during Ang II-mediated HTN and may provide a novel avenue to better understand the role of this mechanosensory pathway in the development of endothelial inflammation in HTN.
Abstract 018: Overexpression Of ERα In Adipose Tissue Contributes To The Anti-contractile Effect Of Mesenteric PVAT By NOX4-induced H
2
O
2
Authors: Costa Tiago J , Fontes Milene T , Webb R Clinton , Wenceslau Camilla F , Enos Reilly T , McCarthy Cam
Published: October 2023

Perivascular adipose tissue (PVAT) induces a significant anti-contractile effect on vasculature, secreting relaxing and constricting factors. It is well established that the PVAT anti-contractile function is lost in cardiometabolic diseases, including obesity. Estrogen receptor alpha (ERα) is known to play a crucial role in the function of adipose tissues and its activation has been associated with adipogenesis by redox balance. In addition, NADPH oxidase 4 (NOX4) is an enzyme that contribute to adipose tissue differentiation and intracellular H
2
O
2
generation. While ERα are expressed in adipose tissue, but their potential role in PVAT function is unknown. To test the hypothesis, adipose tissue specific ERα overexpressing female mice at 30 weeks old were used. We aimed to determine whether the overexpression of ERα would enhance the anti-contractile effect of PVAT by NOX4-induced H
2
O
2
. The high-fat diet (40% fat) was administered for 20 weeks and mesenteric resistance arteries, with and without PVAT, were isolated and used for measurements of vascular function, protein levels (Western blot) and redox signaling (Amplex Red). The higher-fat diet increases the weigh in ~ 80% in Wild Type (WT) and ERα (p>0.05). The anti-contractile effect of PVAT was exacerbated in arteries of ERα mice than WT [Area under the curve (AUC): WT 369±2
vs.
ERα 230±1 p<0.05). NOX1/4 inhibitor (GSK136901) increased the vasoconstriction only in arteries from ERα mice (WT 348 ±4
vs.
ERα 580±5 p<0.05). NOX4 (0.19±0.01 vs 2.3±0.6) and H
2
O
2
(> 70%) levels were higher in PVAT from ERα mice than WT. In addition, 6 uM of H
2
O
2
, concentration corresponding to the observed in PVAT from ERα mice, induced vasodilatation in arteries from WT. On the other hand, a high-fat diet abrogates the anti-contractile effect of PVAT (WT with PVAT 233±1
vs.
without PVAT 241±4) and has partially restored in the ERα, analyzing LogEC
50
(with PVAT 5±0.7 and without PVAT 6±0.8 M). In addition, H
2
O
2
levels (>50%) were higher in PVAT from the high fat diet fed ERα mice, but NOX4 levels did not change. In summary, overexpression of ERα increases H
2
O
2
levels derived from NOX4, contributing to the anti-contractile effect of PVAT in healthy animals. This effect is partially restored in mice on a high-fat diet with overexpressed ERα.
Abstract 028: Sex Differences In Hypertension And Cerebral Oxidative Stress Of Preeclamptic Rat Offspring
Authors: Smith Jonna B , Smith Savanna , Jones Kylie , Castillo Angie , Cunningham Mark W
Published: October 2023

Intrauterine growth restriction (IUGR) occurs when the fetus doesn’t receive adequate nutrients. The leading cause of IUGR is placental ischemia, often seen in preeclampsia (PE). Adult IUGR offspring have an increased risk of HTN and oxidative stress (OS), with sex differences. Studies have shown that IUGR males have HTN, cerebrovascular dysfunction (CVD), and cerebral OS at 17 wks; IUGR females are NT with limited CVD and no significant OS. Our study aims to characterize the sex differences in timing and changes of cerebral OS in IUGR offspring. HTN and OS are both present in adult IUGR males, however, the order and timing is unknown. We hypothesize that OS leads to HTN. To test this, we examined the changes in brain OS in 12 wk IUGR and control (CON) males (before HTN), and 17 wk IUGR males (which have HTN) vs CON males. We investigated the same time points in NT females.

We divided pups from normal pregnant and PE Sprague-Dawley rats into groups based on pregnancy status and sex. Pups were aged to 12 and 17 wks. We collected brains to measure OS via western blot (Manganese superoxide dismutase) and colorimetric assays: hydrogen peroxide (H
2
O
2
) and total antioxidant capacity (TAC).

12 wk IUGR and CON offspring were NT with no changes between groups or sex. There were no changes in cerebral H
2
O
2
, TAC, and MnSOD for IUGR offspring compared to their CON. At 17 wks, IUGR males had elevated mean arterial pressure (MAP; 143.0 ± 1.7 vs 133.0 ± 2.7 mmHg; p < 0.01). IUGR females were NT. Cerebral H2O2 levels in IUGR males were elevated (20.5 ± 8.0 vs 3.7 ± 1.0 uM/ mg Protein; p < 0.05) and unchanged in IUGR females. TAC in IUGR males was decreased (157.7 ±21.9 vs 234.9 ± 22.0 mM Trolox/mg Protein; p < 0.05) and unchanged in IUGR females. MnSOD was elevated in females with no differences between groups (150.4 ± 3.8 vs 110.9 ±7.1 IU/Protein/CON%; p < 0.002).

All 12 wk rats were NT with no changes in cerebral OS. At 17 wks, IUGR males were HTN with elevated cerebral OS, with no changes in females. The lack of increase in OS at 12 wks, suggest that it’s not a precursor to HTN in males, but rather OS is a downstream effect from HTN. Future studies will look at factors that contribute to HTN and CVD in in IUGR offspring. Studies will also examine why 17 wk female IUGR are protected from HTN and cerebral OS.
Abstract 032: Downregulation Of Trpm7 Is Associated With Inflammation And Fibrosis In Aldosterone-induced Hypertension
Authors: Rios Francisco J , Camargo Livia L , Neves Karla B , Alves-Lopes Rheure , Montezano Augusto C , Touyz Rhian M
Published: October 2023

Transient Receptor Potential Melastatin 7 (TRPM7) cation channel is a chanzyme (channel + kinase) that influences cellular Mg
2+
with an important role in cell homeostasis. Here, we investigated the importance of TRPM7-Mg
2+
in inflammatory response and oxidative stress in cardiac fibroblasts (CF) in the context of hypertension and organ damage induced by aldosterone and salt. Wild-type (WT) and TRPM7-deficient (M7+/Δ) mice were treated with aldosterone (600 μg/Kg/day) plus NaCl (1% drinking water), 4 weeks. Blood pressure (BP) was evaluated by tail cuff. Molecular mechanisms were investigated in primary culture of CF. Ca
2+
influx was assessed by fluorescence microscopy. Gene and protein expression was assessed by real-time PCR and immunoblotting respectively. M7+/Δ mice exhibited reduced TRPM7 expression (30%), phospho-TRPM7 (62%) and total tissue [Mg
2+
] (28%). This was recapitulated in WT-aldo-salt. Aldo, salt and aldo-salt increased BP in M7+/Δ mice. In WT, only aldo-salt increased BP (134 mmHg). Cardiac collagen deposition (68%), expression of IL-6, TGFβ, p-Smad3 and p-ERK1/2 were increased (1.5-1.8 fold) in aldo-salt M7+/Δ versus WT. M7+/Δ CF exhibit increased mineralocorticoid target gene SGK1 (3-fold vs WT), which was increased by aldosterone (10-fold vs WT-aldo 5.4-fold). CF M7+/Δ had reduced aldosterone-induced calcium influx (peak-response 105±0.7 vs WT:149±10). CF M7+/Δ exhibited reduced proliferation (30%), and increases protein expression of TGFβ, IL-6, p-Smad3 and p-ERK1/2 (1.4-2.0-fold) vs WT. Nox2 (70%) and Nox4 (120%) expression was increased in CF M7+/Δ vs WT. Aldosterone increased Nox4 (54%) only in CF WT. These effects were associated with increased expression of oxidized peroxiredoxin-3 (76%). Increased gene expression for the antioxidant catalase (1.7-fold), NQO1 (2.6-fold) and Prdx1 (1.7-fold) was increased in M7+/Δ CF. Mg
2+
supplementation normalized intracellular [Mg
2+
], cell proliferation, protein phosphorylation and Nox4 expression in CF TRPM7+/Δ (p<0.05). We identify a novel anti-inflammatory and anti-fibrotic role for TRPM7/Mg
2+
in cardiac fibroblasts. These processes are downregulated in aldo-induced hypertension.
Abstract 034: ROCK2 Specific Inhibition Decreases Th17/Treg Balance And Reduces DOCA-salt Hypertension-associated End-Organ Damage
Authors: Fehrenbach Daniel J , Smart Charles D , Wassenaar Jean W , Madhur Meena S
Published: October 2023

Hypertension is characterized by immune activation and inflammatory end-organ damage, yet there are no therapies for hypertension that specifically target the immune system. We previously demonstrated a critical role for pro-inflammatory CD4+ T cell derived cytokines, interleukin 17A (IL-17A) and IL-21, in the full development of hypertension and hypertension-associated renal and vascular damage. Rho-associated coiled-coil containing protein kinase 2 (ROCK2) has been shown to function as a molecular switch upregulating Th17 and inhibiting anti-inflammatory regulatory T cell (Treg) differentiation in autoimmune diseases. We found that ROCK2 expression is upregulated in splenic and renal CD4+ T cells in a DOCA (deoxycorticosterone acetate)-salt model of hypertension. We hypothesized that hypertensive stimuli increase T cell ROCK2 activity, leading to increased Th17/Treg ratios, and that selective ROCK2 inhibition is a potential novel therapeutic target for hypertension-associated end-organ damage. We showed in vitro that KD025, an orally bioavailable ROCK2 inhibitor, inhibits Th17 proliferation and IL-17A/IL-21 production. We then tested ROCK2 inhibition in vivo where DOCA salt-induced hypertension (uninephrectomy + 100mg DOCA pellet + 1% NaCl drinking water) was initiated in all mice allowing for hypertension to develop for 10 days, followed by treatment with KD025 (50mg/kg daily, n=11) or vehicle (n=8) starting on day 11 through the remainder of the protocol. KD025 treatment significantly attenuated cardiac hypertrophy (8.7±0.3 vs 7.6±0.2 mg/grams body weight, p=0.005) and left ventricular (LV) fibrosis (4.9±1.7 vs 2.4±1.3% LV area, p=0.008). Flow cytometric analysis revealed that KD025 decreased F4/80+ infiltrating cardiac macrophages (24,000±578 vs 18,918±1,430 cells, p=0.011) and decreased the Th17/Treg ratio (0.251±0.041 vs 0.117±0.009, p=0.003) in the heart. Furthermore, KD025 attenuated renal infiltration of CD3+ T cells (59.0±5.6 vs 43.5±3.8 cells/mg tissue, p=0.019) and albuminuria (163.4±24.1 vs 103.1±21.9 ug albumin/mg creatinine, p=0.011). These data indicate that the ROCK2-specific inhibitor KD025 may be a novel therapeutic for the treatment of hypertension and the associated end-organ damage.
Abstract 039: B-arrestin Defficiency In The Subfornical Organ Alters Blood Pressure Regulation
Authors: Mathieu Natalia , Nakagawa Pablo , Muskus Veitia Patricia , Brozoski Daniel , Grobe Justin L , Sigmund Curt D
Published: October 2023

The activation of the angiotensin type 1 receptor (AT
1
R) in the brain through the canonical Gαq-mediated signaling results in dipsogenic and pressor responses to angiotensin (AngII). Non-canonical or β-arrestin-mediated signaling of AT
1
R activation is hypothesized to counterbalance maladaptive G protein signaling during disease. Recently, we found that global genetic deletion of β-arrestin2 (ARRB2) resulted in an exacerbated increases in blood pressure (BP) in response to deoxycorticosterone acetate-salt treatment, suggesting a protective role for ARRB2 in hypertension. We hypothesized that ARRB2 activation specifically within the brain contributes to BP regulation. First, global male and female ARRB2-deficient (ARRB2-KO) mice were used to evaluate acute BP responses to intracerebroventricular (ICV) infusion of AngII (1μg). Although no differences at baseline BP were observed, ARRB2-KO mice exhibited greater maximal BP after AngII (144±17 vs. 177±24 mmHg; n=14 and 19, p<0.05). Next, we examined the role of ARRB2 within the brain via ICV injection of adeno-associated virus (AAV)-CRE into ARRB2
FLOX
mice. Infection with ICV AAV-CRE primarily targeted the subfornical organ with minimal off-targets. In preliminary studies, these mice exhibited higher BP in response to AngII (179±5 vs. 187±6 mmHg; n=4 p=0.09). Thus, deletion of ARRB2 within the subfornical organ exacerbates the pressor response to Ang II. Finally, we evaluated BP responses to ICV infusion of a β-arrestin biased agonist specific for the AT
1
R (TRV120027 0.5 ug/h) in spontaneously hypertensive BPH/2J and normotensive BPN/3J mice. BPH/2J mice exhibited significantly higher baseline BP compared to BPN/3J (124±5 vs. 153±3 mmHg; n=4 p<0.05), and greater reductions in BP with ICV infusion of TRV120027 (-4±3 vs. -9±1 mmHg; n=4 p<0.05), suggesting that the beneficial BP-lowering effects of β-arrestin activation are more robust during hypertension. Overall, these data indicate that ARRB2 in the subfornical organ is protective against increases in BP. Stimulation of the AT
1
R β-arrestin axis in the brain may represent a novel strategy to treat hypertension.
Abstract 044: Knocking Out Sodium Glucose-Linked Transporter 5 Prevents Fructose-Induced Salt-Sensitive Hypertension
Authors: Forester Beau R , Brostek Autumn , Gonzalez-Vicente Agustin , Garvin Jeffrey
Published: October 2023

We showed: 1) a 20% fructose high-salt (3.7% NaCl; FHS) but not a 20% glucose high-salt diet increases systolic blood pressure (SBP) in a sex independent manner; 2) on normal salt, 20% fructose augments the ability of angiotensin II (Ang-II) to increase proximal tubule superoxide (O
2
-
); and 3) rat proximal tubule brush borders express sodium glucose-linked transporters (SGLT) 4 and SGLT5, which transport fructose. However, whether FHS enhances Ang II-stimulated proximal tubule O
2
-
, its role in FISSH and the roles of SGLT4 and 5 are unknown. We hypothesized that fructose transport mediated by SGLT5 but not SGLT4 is required for FHS to enhance Ang II-stimulated proximal tubule O
2
-
in males and females, and this is required for fructose-induced salt-sensitive hypertension (FISSH). We measured SBP in wild-type, SGLT4 knockout, and SGLT5 knockout male and female rats on a Sprague Dawley background fed FHS for 7 days. We also measured basal and Ang II-stimulated (37 nM) O
2
-
in proximal tubule suspensions from these rats using the lucigenin assay. FHS increased SBP in wild type male rats 11 ± 2 mmHg (from 131 ± 4 to 142 ± 5 mmHg; n = 5, p < 0.01) and in female rats by 21 ± 5 mmHg (from 114 ± 4 to 130 ± 3 mmHg; n = 8; p < 0.01), not significantly different from males. FHS increased SBP in SGLT4 males by 22 ± 3 mmHg (from 126 ± 5 to 149 ± 6 mmHg; n = 8, p < 0.01) and in females 21 ± 5 mmHg (from 126 ± 3 to 138 ± 2 mmHg; n = 8; p < 0.01), not significantly different from males. In SGLT5 males SBP was 128 ± 6 on day 0 and 129 ± 8 mmHg on day 7 (n = 7). In SGLT5 females SBP was 114 ± 4 on day 0 and 117 ± 5 mmHg on day 7 (n = 7). Ang-II increased O
2
-
by 11 ± 3 relative light units (RLU)/μg/s from 53 ± 9 to 64 ± 8 RLU/μg/s (n = 11, p < 0.01) in tubules from wild type males fed FHS for 7 days and by 10±3 RLU (n = 5; p < 0.02) in females fed FHS for 7 days. Ang-II increased O
2
-
by 8 ± 2 RLU/μg/s in proximal tubules from FHS-fed SGLT4 males (n = 6, p < 0.02) and by 8 ± 2 in FHS-fed females (n = 6, p < 0.01). Ang-II did not significantly stimulate O
2
-
in proximal tubules from FHS-fed SGTL5 male or female rats (Δ 3±7 and 0±3 RLU/μg/s, n = 5 and 3, respectively). We conclude that transport by SGLT 5 but not SGLT4 is required for FHS to augment Ang II-stimulated proximal tubule O
2
-
in males and females, and that the increase in proximal tubule oxidative stress is required for FISSH.
Abstract 057: The Positive Effect Of Renal Denervation Persists Even Through Regrowth Of Afferent Axons Improving Renal Afferent Nerve Activity During High Sodium Intake In Rats
Authors: Rodionova Kristina , Ditting Tilmann , Lindner Sophia , Linz Peter R , Cordasic Nada , Hilgers Karl F , Schiffer Mario , Schmieder Roland E , Amann Kerstin , Veelken Roland
Published: October 2023

Introduction:
Previous work of ours suggests that pathologically decreased sensitivity of renal afferent neurons (reduced number of highly active tonic neurons) due to high salt diet is normalized 1 wk after renal denervation (DNX). Now we tested the hypothesis that normalized sensitivity of renal neurons persists after DNX even through regrowth of afferent axons after 12 wk. This morphological regrowth of afferent nerves 12wk after renal DNX (postDNX) was shown by us previously.

Material and methods:
6 male Sprague Dawley (SD) rats were put on high salt diet (HS; 8% NaCl) for 10 days. In another group of 12 rats on high salt diet (HS) left kidneys were denervated (postDNX) 12 weeks prior to examination. Rats on standard diet were used as controls. Harvested dorsal root ganglion neurons (DRG Th11-L2) with renal afferents were investigated in primary neuronal cell culture using current clamp mode to assess action potential generation during current injection and to characterize neurons as tonic highly active and phasic less active neurons.

Results:
In renal neurons from rats on HS the relation of tonic to phasic neurons shifted towards less active phasic units (62% tonic neurons in controls [n=72 out of 114] vs. 42% [n=37 out of 86] on HS, (p<0.05, z-test)). Denervation of the left kidney in rats on high salt diet (HS-DNX) led to a recovery of afferent renal DRG neurons after 1 wk; (40% tonic neurons [n=32 out of 80] on HS vs. 72% tonic neurons [n=58 out of 81] on HS+DNX)(p<0.05, z-test)). Even 12 weeks after renal denervation this regained electrophysiological property of tonic firing persisted (43% tonic neurons [n=35 out of 82] on HS vs. 69% tonic neurons [n=46 of 67]on HS-postDNX, p<0.05, z-test)).

Conclusion:
The reduced proportion of highly active tonic neurons increased 1 wk after renal denervation to control levels. Even 12 weeks after renal denervation, this effect of renal denervation persisted, and the sensitivity of renal neurons to electrical stimuli did not change despite high-salt diet. Hence, the positive effects of renal denervation persisted even through the regrowth of renal afferent axons to the kidney.
Abstract 076: The Blood Pressure-associated Non-coding Single Nucleotide Polymorphism Rs28451064 Influences Gene Expression In Human Endothelial Cells
Authors: Pandey Rajan , Liu Yong , Liu Pengyuan , Therani Bhavika , Geurts Aron M , Liang Mingyu
Published: October 2023

Most Blood Pressure (BP)-associated single nucleotide polymorphisms (SNPs) are in intronic or intergenic regions. For the noncoding SNP rs28451064, allele A is associated with elevated diastolic BP as well as increased risk of myocardial infarction and angina pectoris. rs28451064 is mapped to the nearby protein coding gene
KCNE2
and is an expression quantitative trait locus for
RCAN1
. We performed a series of studies to experimentally identify genes influenced by rs28451064 and to begin to understand the mechanism. We differentiated a human induced pluripotent stem cell (iPSC) line to endothelial cells (iECs). The iECs were purified by sorting with VE-cadherin. Chromatin conformation analysis using Micro-C identified several genes including
MRPS6
and
SLC5A3
as sharing a chromatin loop with rs28451064. We used CRISPR-Cas9 to generate isogenic iPSC lines with either homozygous low BP (G) or high BP allele (A) of rs28451064. Three clones of iPSCs with each genotype were established by screening several hundred clones. The edited iPSCs were differentiated into iECs for RNA-seq analysis, which showed that several genes that are genomically close to rs28451064 tended to be differentially expressed, including
KCNE2
,
SMIM34
, and
RCAN1
. The edited iPSC lines were subjected to six rounds of differentiation to iECs for targeted real-time PCR analysis of genes near rs28451064, including the genes mentioned above. The results indicated significantly increased expression of
SMIMI11
(1.93±0.25 fold) and
SMIM34
(1.45±0.13 fold) (n=34) and decreased expression of
RCAN1
(0.90 ± 0.03 fold)
and SLC5A3
(0.90 ± 0.03 fold) (n=36) in iECs with the high BP allele compared to the low BP allele of rs28451064 (p<0.05).
SMIM11
and
SMIM34
encode small integral membrane proteins with unknown functions.
RCAN1
encodes regulator of calcineurin 1, and
SLC5A3
encodes sodium/myo-inositol cotransporter 1. Interestingly, the closest genes,
MRPS6
and
KCNE2
, along with
MRPL23
, a control gene on a different chromosome, were not significantly differentially expressed. In summary, we have shown definitively that rs28451064 influences the expression of several local genes in an allele-specific manner. The results provide new insights into the function of a BP-associated noncoding SNP.
Abstract 082: Dysregulated Microbiota-indole Axis In Male, Not Female Ace2 Deficient Salt-sensitive Hypertension
Authors: Bardhan Pritam , Mei Xue , Lai Khanh , Mell Blair , Tummala Ramakumar , Atluri Rohit , Kyoung Jun , Joe Bina , Yang Tao
Published: October 2023

Introduction:
Aberrant tryptophan (Trp) absorption may promote hypertension (HTN) due to gut microbiota converting Trp to pro-hypertensive indoles. Intestinal
Ace2
regulates Trp homeostasis, but its role in salt-sensitive HTN is unknown. Further, lack of
Ace2
can affect males and females differently because
Ace2
is transcribed from the X-chromosome in a genomic region which escapes X-inactivation. Therefore, we hypothesized that sex-specific intestinal ACE2 dependent transport of Trp contributes to salt-sensitive HTN via the microbiota-indole axis.

Methodology:
8-10-week-old Dahl salt-sensitive (S) rats and newly generated CRISPR/Cas9
Ace2
deficient S rats of both sexes (N= 7 rats/group) were maintained on either a low (0.3 % NaCl, LS) or high (2 % NaCl, HS) salt diet for 21 days. BP was measured by radiotelemetry for 24 hours on day 21. Serum and cecal content were collected for indole measurement and microbiota analysis.

Results:
Mean arterial BP of S
Ace2
deficient rats on LS was elevated in females by 38 mmHg (S

Ace2
-/-

170 ± 10 vs. S 132 ± 3;
P
=0.0028) and in males by 18 mmHg (S

Ace2
-/Y

140 ± 6 vs. S 122 ± 1.5;
P
=0.01), and further elevated on HS by 68 mmHg in females (S

Ace2-/-

211 ± 14 vs. S 143 ± 4.5;
P
=0.0003) and by 40 mmHg in males (S

Ace2-/Y

182 ± 9 vs. S 142 ± 3;
P
=0.0009). These were associated with elevated serum indole levels mainly in male S

Ace2
-/Y

rats, compared to S rats on both LS (1.2 ± 0.2 vs. 0.3 ± 0.2 μM;
P
=0.02) and HS (1.5 ± 0.25 vs. 0.2 ± 0.03 μM;
P
=0.0001). Microbiota analysis revealed (1) significant shifts in β-diversity in
Ace2
deficient rats of both sexes with a higher R value and a lower P value in HS males; (2)
Ace2
deficiency resulted in higher abundance of Trp-utilizing, indole-producing bacteria
Escherichia
-
Shigella
in males on both LS (
P
<0.05) and HS (
P
<0.0001). Strikingly, none of these differences were found in females.

Conclusion:
Lack of
Ace2
led to increased mean BP regardless of sex and salt intake. In males,
Ace2
deficiency led to increased indole levels with higher abundance of
Escherichia
-
Shigella
. Overall microbial changes by salt and ACE2 deficiency were found mainly in males, suggesting a more resilient microbiota in females. Thus, the microbiota-indole axis may be a potential therapeutic target for BP control in males with altered Trp metabolism.
Abstract 105: Potential Effect Modifiers In The Relationship Between Late-life Systolic Blood Pressure And Alzheimer’s Neuropathology
Authors: Cai Wenjie , Ma Yuan , Blacker Deborah , Neitzel Julia
Published: October 2023

Mixed evidence exists on the relationship between late-life blood pressure (BP) and Alzheimer’s disease (AD) pathology such as amyloid and tau deposition in the brain, possibly partly due to heterogeneous effects across population subgroups. Utilizing data from the National Alzheimer’s Coordinating Center longitudinal dataset, we investigated to what extent age (<65 yrs, 65-74 yrs, >74 yrs),
APOE
genotype, stroke history, and baseline cognitive status modified the association between systolic BP (SBP) at baseline and amyloid and tau neuropathology in 3165 older adults (mean [SD] age=74 [9.8] yrs, women 47.4% [1501 of 3165]). Amyloid pathology was assessed by Thal staging, diffuse plaque, neuritic plaque and tau pathology was assessed by Braak staging at brain autopsy. Effect modification was assessed by stratified analyses and interaction analyses in multinomial logistic regression with adjustment for age at first visit, sex,
APOE
genotype, education years and antihypertensive medications use. We observed that both
APOE e4
carriership (interaction p-value = 0.03) and stroke history (interaction p-value = 0.03) statistically modified the association of SBP with amyloid pathology, whereas baseline cognitive status modified the association with both amyloid and tau pathology (interaction p-value <0.001). The SBP-amyloid association appears to be pronounced in older age group (65-74 group: OR=1.19 [CI:1.05-1.35], >74 group: OR=1.12 [CI:1.02-1.23]),
APOE e4
carriers (OR=1.17[CI:1.02-1.34]), and those with stroke history (OR=1.69 [CI:1.20-2.38]) and participants with dementia at baseline (OR=1.16 [CI:1.05-1.27]), whereas SBP-tauopathy association was only found in
APOE e4
carriers (OR=1.25[CI:1.05-1.48]), participants without stroke history (OR=1.12[CI:1.01-1.24]), and participants with normal cognitive function at baseline (OR=1.67[CI:1.24-2.24]). Our study suggests that the relationship between SBP and amyloid and tau pathology may vary across subgroups based on age,
APOE
genotype, cerebrovascular burden, and stage of cognitive impairment. The underlying mechanisms warrant further investigation to inform precision blood pressure control for the prevention of dementia.
Abstract 114: Effect Of Dose Of Chlorthalidone And Hydrochlorothiazide Dosing On Cardiovascular Outcomes In The Diuretic Comparison Project (DCP)
Authors: Hau Cynthia , Ishani AREEF , Glassman Peter , Taylor Addison A , Ferguson Ryan , Leatherman Sarah , Cushman William C
Published: October 2023

Background:
The Diuretic Comparison Project (DCP) showed chlorthalidone (CTD) to be comparable with hydrochlorothiazide (HCTZ) in cardiovascular prevention among older hypertensive patients. Previous hypertension outcome trials showing benefit for either, higher doses of each were predominant. DCP randomized 13,523 participants to stay on existing HCTZ (25/50mg) or switch to a dose equivalent CTD (12.5/25mg). Such trial data was used to test if CTD was superior to HCTZ when prescribed at a higher dose.

Methods:
This analysis examined treatment effects with respect to low (25mg HCTZ/12.5mg CTD) and high (50mg HCTZ/25mg CTD) dose regimens. Outcomes including non-fatal MI, stroke, acute heart failure, revascularization for unstable angina and non-cancer death were tested individually and as a composite measure.

Results:
There were no differences for the composite and all individual outcomes in both low dose (12,781/13,523; 94.5%) and high dose (742/13,523; 5.5%) groups. Mean age (72 y), baseline blood pressure (139 mm Hg), and follow-up time (2.4 y) were similar across doses. High dose participants had a greater prevalence of diabetes and received more antihypertensive drugs. Consistent with DCP results, there was a significant interaction between treatment group and history of MI/stroke in the low dose group (p=0.002); this was not found in the high dose group (p=0.8).

Conclusion:
As in the main DCP trial results, there were no differences in outcomes between CTD and HCTZ within higher or lower dose subgroups. Treatment interaction with prior mi/stroke was found with lower doses. However, the high dose group was small and might not reflect a true treatment effect at higher doses.