bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 419: Investigating the Role of Epidermal Growth Factor Receptors in Angiotensin II-induced Hypertrophic Signalling in Cardiomyocytes
Authors: Wang Zhen , Chan Hsiu-Wen , Chen Chen , Thomas Walter G , Paravicini Tamara M 
Published: March 2022

Cardiac hypertrophy is characterized by stimulation of hypertrophic signalling pathways representing the reactivation of the ‘foetal gene program’ (hypertrophic genes) and activation of the MAPK pathway. Angiotensin II (AngII) stimulates cardiac hypertrophy, a process which may involve the transactivation of epidermal growth factor receptors (ErbBs). Three subtypes of ErbBs are present in the postnatal heart (ErbB1, 2, 4), yet their precise roles are poorly understood. We have shown that the ErbB4-selective ligand neuregulin 1 beta-1 (NRG1-β1) potently stimulates cardiomyocyte hypertrophy. We now aim to determine if AngII activates hypertrophic signalling in cardiomyocytes by transactivation of ErbBs, particularly ErbB4.
Ventricular cardiomyocytes from neonatal rats were treated with AngII (100 nM) or NRG1-β1 (10 nM) to induce hypertrophic signalling. ErbB1, 2, 4 were downregulated using shRNA and siRNA. Reporter assays were used to measure promoter activity of the hypertrophic genes myosin light chain 2v (MLC-2V), atrial natriuretic peptide (ANP) and cyclin D. ERK1/2 activation was measured via Western blot.
NRG caused robust activation of the hypertrophic gene MLC-2V (11.3 fold increase versus untreated control) that was significantly reduced to 4.3-fold by silencing of ErbB4 (n = 5, P < 0.01), but not ErbB1 or ErbB2, confirming that ErbB4 activation contributes to hypertrophic signalling. However, knockdown of ErbB1, 2 or 4 did not affect AngII-induced activation of MLC-2V (5.8 fold increase versus untreated control). Similar results were seen for ANP and cyclin D. Similarly, ErbB4 knockdown reduced NRG1-β1-induced MAPK activation from 21.0-fold to 13.5 fold (n=4, p<0.05), whilst Ang II-induced MAPK activation was unaffected.

Cardiomyocyte hypertrophy can be promoted by activation of AT
1
and ErbB4 receptors. AngII-induced hypertrophic signalling (measured by promoter activity of MLC-2V, ANP and cyclin D and MAPK activation) may not involve transactivation of ErbB receptors. We are now extending these findings
in vivo;
we have established a model of cardiac-specific ErbB4 deletion using Cre/Lox recombination, and are undertaking echocardiographic analysis to measure cardiac hypertrophy and function.
Abstract 464: Molecular and Functional Interaction of the Angiotensin AT2-Receptor and the Receptor MAS
Authors: Villela Daniel , Leonhardt Julia , Muenter Lisa M , Lucht Kristin , Benz Verena , Namsolleck Pawel , Alenina Natalia , Unger Thomas , Bader Michael , Santos Robson A , Steckelings Ulrike M 
Published: March 2022

The angiotensin AT2-receptor (AT2R) and the receptor MAS both mediate very similar tissue-protective actions. Furthermore, AT2R-antagonists have been reported to block effects of the MAS-agonist Ang 1-7. Therefore we speculated that a molecular association between these two receptors exists resulting in functional interactions.
Fluorescence resonance energy transfer (FRET) with photobleaching was used for detection of a putative interaction between MAS and AT2R in HEK-293 cells transfected with vectors encoding MAS or AT2R fused in the C-terminus with CFP or YFP. In addition, functional interaction of AT2R and MAS was assessed in primary mouse astrocytes by determining the effect of respective agonists and antagonists on expression of mRNA encoding the chemokine receptor CX3CR1.

There was significant FRET efficiency of 10.8±0.8% when AT2-YFP and MAS-CFP were co-expressed indicating heterodimerisation. Both, MAS and AT2R also formed homodimers (FRET efficiencies: 7.4±0.8% or 9.2±0.8%, respectively). No FRET efficiency was observed with an unrelated transmembrane receptor, and expression of non-fluorescent MAS and AT2R competed with FRET efficiencies, which both supports specificity of receptor interactions. Regarding putative functional interactions, AT2R-agonist induced increases in CX3CR1 mRNA expression were absent in astrocytes concomitantly treated with the MAS-antagonists A-779 or in Mas-deficient cells. Likewise, effects of the Mas agonist Ang 1-7 were undetectable in cells concomitantly treated with the AT2-antagonist
PD123319
or in AT2-deficient cells.

This study provides evidence that MAS and AT2 receptors form homo- and heterodimers. Heterodimerisation may be of functional relevance, because in primary mouse astrocytes, Mas and AT2 seemed only functional in the presence of the respective other receptor.
Abstract 604: Renin Independent Ang-(1-12) Production in both the Systemic and Coronary Circulation
Authors: Moniwa Norihito , Varagic Jasmina , Nagata Sayaka , Ahmad Sarfaraz , Simington Steven W , VonCannon Jessica L , Shimamura Yoshinosuke , Hasegawa Koichi , Takizawa Hideki , Ura Nobuyuki , Tanaka Shigemichi , Furuhashi Masato , Yoshida Hideaki , Miura Tetsuji , Ferrario Carlos M 
Published: March 2022

Although
angiotensin-(1-12) [Ang-(1-12)] is recognized as an alternate precursor for Ang

II, the biochemical pathways by which the substrate is released from
angiotensinogen (Aogen) are not yet described.
We examined the role of renin on Ang-(1-12) formation from synthetic icosapeptide
[Ang-(1-20)] in the systemic and coronary circulation of Wistar Kyoto rats
(WKY). In experiment 1, rats underwent bilateral
nephrectomy (BNX) or sham operation.
After 48 hours, rats were assigned to sham+saline, sham+Ang-(1-20), or
BNX+Ang-(1-20). Ang-(1-20) (20 nmol/kg/min)
or saline was infused for 15 minutes. At
the end of the study blood was collected for measuring Ang-(1-12), Ang I and
Ang II by RIA. Ang-(1-20) infusion increased
arterial pressure in both sham and BNX compared to sham+saline (P<0.001). Plasma Ang-(1-12), Ang I, and Ang II were significantly
higher in sham+Ang-(1-20) compared to sham+saline. BNX+Ang-(1-20) showed significant increase in
Ang I and a tendency for augmented Ang-(1-12) and Ang II compared to sham+saline
(Figure). In experiment 2, the hearts extracted
from WKY were mounted on Langendorff apparatus.
After recovery, 10 nmol or 100 nmol of Ang-(1-20), or 100 nmol of
Ang-(1-20) with WFML-1 (a rat renin inhibitor) were perfused in the coronary
circulation. Buffer was recirculated
after addition of Ang-(1-20) and WFML-1.
Perfusate was collected at 0, 5, 15, 30, and 60 min for measurement of angiotensins
by RIA. The 100 nmol of Ang-(1-20)
increase Ang-(1-12), Ang I, and Ang II compared to 10 nmol both in the absence
and in the presence of WFML-1 (Figure).
We conclude that Ang-(1-12) is cleaved from Aogen by an enzyme that is
not renin in both systemic and coronary circulation.
Abstract 609: Potential Role Of Adipose Derived Stem Cells In Attenuating Pulmonary Hypertension
Authors: Rathinasabapathy Anandharajan , Shenoy Vinayak , Bruce Erin , Espejo Andrew , Raizada Mohan , Katovich Michael 
Published: March 2022

Pulmonary hypertension (PH) is a devastating cardiopulmonary
disorder, which often results in right heart failure. In spite of recent
advancements in the pharmacotherapy of pulmonary hypertension, the mortality
rate has not been significantly reduced. Recently stem cell therapy has gained
more attention for the treatment of cardiopulmonary diseases. Despite the
promising results shown by bone marrow derived stem cells in attenuating right heart
failure in PH, it remains a highly invasive and laborious procedure. On the contrary, adipose derived stem cell
(ADSC) presents a less complicated option. Therefore, we proposed to use ADSC in
attenuating the progression of PH. Monocrotaline
(MCT) at a 50 mg/kg dose was injected to eight-week old male Sprague-Dawley
rats to induce PH. Four weeks post MCT injection; animals developed PH with
elevated right ventricular systolic pressure (RVSP, Control: 28.65±0.40; MCT:
78.06±6.4 mmHg; n=6-8), and right ventricular hypertrophy (RVH, Control;
0.22±0.005; MCT: 0.48±0.054 [ratio, AU]; n=6-8). ADSC (isolated from inguinal fat of healthy male

rat) were characterized using flow cytometry (CD44
+
, CD90
+
,

CD34
-
and CD45
-
) and their differentiation to adipocyte

was confirmed by Oil Red O staining. ADSCs

(1x10
6
; passage 2-4) were injected through the jugular vein, 14 days

post MCT injection. After 2 weeks following
ADSC therapy, RVSP (MCT+ADSC: 55.67±8.99 mmHg; n=6-8) and RVH (MCT+ADSC:
0.32±0.062 [AU]; n=6-8) was significantly reduced. Moreover, the elevated right ventricular end
diastolic pressure (RVEDP) and ±dP/dT associated with PH were attenuated
significantly in ADSC treated MCT animals. An additional group of animals received only

the ADSC conditioned media (CM) from 1x10
6
cells. After 2 weeks of CM administration, RVSP

(MCT+CM: 42.11±4.66 mmHg; n=5) and RVH (MCT+CM: 0.29±0.020 [AU]; n=5) was
significantly reduced. Further, the RVEDP
and ± dP/dT were also significantly reduced in CM treated MCT animals. Collectively, either ADSC or conditioned media
alone can attenuate the progression of PH, which suggest that paracrine-like substances
may be responsible for the beneficial effects observed and may be considered as
a potential therapeutic alternative for the treatment of right heart failure.
Abstract 615: Progression Of Atherosclerosis Plaque Area In Postmenopausal Women During Controlled Traditional Risk Factors
Authors: Perez Hernan A , Garcia Nestor H , Spence J. David , Armando Luis J 
Published: March 2022

The onset of symptoms of coronary heart disease in women are displaced a decade compared with men after menopause, but currently there are reports in which 1 in 4 women is less than 59 years, suggesting that in the pre-menopausal period there is marked progression of vascular disease. Therefore we evaluated the presence of atherosclerotic disease in primary care menopausal women. Additionally, we hypothesized that the strict control of the traditional risk factors is not enough to prevent vascular disease progression.
Methods, post-menopausal women without cardiovascular events from Argentina were evaluated. Framingham risk score with body mass index (FRSbmi) and carotid ultrasound were employed for the evaluation. Carotid total plaque area (TPA) by ultrasound was determined as previously described by Spence et al (Stroke 2002;33;2916-2922). Additionally, LDL cholesterol (LDL), triglycerides (Trig) and HbA1c were measured. All determinations were done on day 1 and after 8 months of treatment. Only patients who achieved all the therapeutic goals (blood pressure (BP)<130/80mmHg, LDL<100mg/dl, Trig<140mg/dl, HbA1c5mm2, stable group, TPA changed 5mm2. In the presence of controlled of traditional risk factors, 50% of the population continued to progressive atherosclerotic disease (Table).

Conclusion, these results indicate that in the presence of controlled traditional risk factors, atherosclerosis may progress in some patients, including in menopausal women. Additional promoters of atherosclerosis should be evaluated in these progressive patients or possibly more intensive treatment should be used in such patients.
Abstract 640: Role Of Endothelial No Synthase Expression On Endothelial Dysfunction In Hypertensive Patients
Authors: Danesh Shahab M , Solow Elizabeth B , Arbique Debbie , Wang Zhongyun , Onwuegbuchu Erica , Mineo Chieko , Vongpatanasin Wanpen 
Published: March 2022

Previous studies have implicated impairment
in the expression of endothelial NO synthase (eNOS) and the S1177 phosphorylated
form of eNOS in mediating endothelial dysfunction in the spontaneously
hypertensive rats. However, the relevance of this observation in human
hypertension remains unknown. Accordingly, we assessed endothelial function in 19
patients with untreated essential hypertension (HTN) and 18 age-matched
normotensive controls (NT), using noninvasive flow mediated vasodilation (FMD)
in the brachial artery. We also assessed endothelial cell (EC) protein
expression of eNOS and S1177 phosphorylated eNOS using immunofluorescence staining of the ECs extracted via J wire
insertion into antecubital veins. We found that FMD was significantly reduced in
the HTN versus the NT group (6.67±0.53 % vs. 9.16±0.98
%, p < 0.05). Despite impairment in endothelial function, we found that
protein expression of total eNOS and S1177 phosphorylated eNOS were not
significantly different between the HTN and NT groups (0.46±0.07 vs.
0.38±0.05 and ±0.05 vs. 0.36±0.08, p = NS,
respectively). In conclusion, our study suggests that impairment in the endothelial
vasodilator function is independent of
eNOS or S1177 phosphorylation of eNOS protein
expression in human hypertension. Further studies are needed to clarify the
molecular basis of endothelial dysfunction in hypertensive patients.
Abstract 68: Mineralocorticoid Receptor Activation Leads to Ocular Choroidal Vessels Dilation and Leakage in Rats and Human Central Serous Chorioretinopathy
Authors: Jaisser Frederic , Zhao Min , Farman Nicolette , Behar-Cohen Francine 
Published: March 2022

We have shown that several cell types of the rat retina express the mineralocorticoid receptor (MR) and the MR-protecting enzyme HSD2 (Zhao et al, FASEB J 2010, 24:3405), raising the question of the role of the MR in normal and diseased retina. We now questionned the role of MR in the choroid vessels lying in the outermost posterior part of the eye. Intravitreous injection of the MR ligand aldosterone in rat eye induced, 24 hrs later, dilation of choroid vessels and fluid leakage, through specific MR activation. Corticosterone (at high doses, not low doses) elicited the same effect, presumably via MR occupancy. We identified an underlying mechanism: aldosterone up-regulates the endothelial vasodilatory potassium channel KCa2.3; its blockade(by intravitreous apamine) prevents aldosterone-induced thickening.
We seached whether human retinal diseases may recapitulate some of the features of MR overactivation. Central serous chorioretinopathy (CSCR) is a vision-threatening eye disease affecting young stressed men, with unknown pathogeny and no validated treatment. Subretinal fluid accumulation originates from dilation and leakage of choroid vessels underneath the retina. Recurrence or chronic evolution may occur, leading eventually to vision loss. Based on aggravation of CSCR by glucocorticoids , on identification of systemic and/or locoregional glucocorticoids as known risk factors, and on the recognized MR affinity for glucocorticoids, we hypothesized that inappropriate/excessive occupancy of the MR by glucocorticoids may occur in the choroid of CSCR patients. Four patients with chronic non-resolved CSCR (and reduction of visual acuity) received low doses of oral eplerenone for 5 weeks. Retinal detachment and choroidal vasodilation were rapidly (1- 2 weeks) resolved; this impressive recovery was associated with improved visual acuity. The benefit was maintained after eplerenone withdrawal. Controlled clinical trials are now required to assess this novel therapeutic approach.
In conclusion, our results identify MR signaling as a novel pathway controlling choroidal vascular bed relaxation and provide a pathogenic link with human CSCR, leading to proposal of short-term MR blockade to reverse the choroid vasculopathy and improve vision.
Abstract 017: Spironolactone Improves Insulin Secretion In Metabolic Syndrome Subjects
Authors: Luther James M , Byrne Loretta M , Yu Chang 
Published: March 2022

Renin-angiotensin-aldosterone system blockade reduces the incidence of type 2 diabetes in clinical trials. We previously demonstrated that aldosterone impairs insulin secretion via a direct beta-cell effect in mice. To test the hypothesis that aldosterone and the mineralocorticoid receptor contribute to impaired insulin secretion in humans, we enrolled subjects with metabolic syndrome treated with hydrochlorothiazide (HCTZ) 12.5 mg daily for 4 weeks (baseline) followed by either Spironolactone 25 mg daily (SPL, n=16) versus Aliskiren 150 mg daily (ALI, n=19) for 4 weeks. We assessed insulin secretion and sensitivity using hyperglycemic clamps during sodium controlled diet (160 mmol Na).
During HCTZ alone, plasma aldosterone correlated inversely with the initial insulin response and disposition index (Figure A), but not with insulin sensitivity index. Plasma aldosterone decreased during aliskiren (-1.5±0.78 ng/dL) and increased during spironolactone treatment (+4.0±0.8 ng/dL; p<0.001 between treatment). Blood pressure, serum potassium, and insulin sensitivity index were unchanged. The acute insulin response increased compared to baseline during SPL compared to ALI (+11.6±7.9 vs -10.7±7.7 μU/mL; p=0.048), whereas insulin sensitivity index was unchanged.
We conclude that aldosterone is inversely associated with the initial phase insulin secretory response, the earliest detectable change in impaired glucose tolerance. Treatment with spironolactone could improve glucose homeostasis by restoring insulin secretion in subjects with metabolic syndrome.
Abstract 027: T cell-Dependent B cell Activation Plays a Role in Mediating Hypertension and Pathophysiology in Response to CD4+ T cells from Reduced Uterine Perfusion Pregnant Rats
Authors: Cornelius Denise C , Castillo Javier , Porter Justin , Campbell Nathan , Thomas Alexia , Amaral Lorena M , Wallace Kedra , Moseley Janae , LaMarca Babbette 
Published: March 2022

Preeclampsia, (PE) newly developed hypertension during pregnancy, is associated with altered immune activation. We have previously shown that adoptive transfer of CD4+T cells from a rat model of PE, the RUPP Rat, increases blood pressure, endothelin (ET) expression, oxidative stress (ROS), inflammation, and agonist autoantibodies to the AT1 receptor in NP recipients, all of which have been shown to play a role in hypertension during pregnancy. The objective of this study was to determine if blockade of lymphocyte crosstalk via the CD40-CD40 ligand interaction between RUPP CD4+ T cells with endogenous B cells in NP recipient rats would improve pathophysiology observed previously with RUPP CD4+ T cells. To do so splenic CD4+ T lymphocytes were magnetically separated, incubated with 2.5 μg/mL anti-CD40 ligand (αCD40L) overnight and transferred into Normal Pregnant (NP) rats on day 12 of gestation (NP+ αCD40L RUPP T cells). On day 19 of gestation, blood pressure (MAP), blood, and tissues were collected from all groups of rats.
MAP was 99+/-1.2 in NP (n=18), 120+/-2.3 mmHg in control RUPP rats (n=19;P<0.001); 121+/-1.2 in NP+RUPP T cells(n=5;P<0.001); but only increased to 103 +/-1.7 in NP + αCD40L RUPP T cells (n=21) (P<0.001 vs NP+RUPP T cells). Placental ROS increased from 160.4 +/- 31 in NP to 380.5 +/- 60.7 in RUPP (p<0.05) and 318.6 +/-89 in NP+RUPP T cells. CD40 ligand binding reduced placental ROS to 118.7 +/-24 in NP + αCD40L RUPP T cells (p<0.05). Plasma IL-6 increased from 38.71+/-8 in NP to 92.94+/-15 in RUPP (p<0.05) and 107.4+/-26 in NP+RUPP T cells and only increased to 50.1+/-3.6 pg/ml in NP + αCD40L RUPP T cells. TNF-alpha increased from 10.9+/-5 in NP to 79+/-15 in RUPP (p<0.05) and 23+/-17 in NP+RUPP T cells and was only 8+/-3 pg/ml in NP + αCD40L RUPP T cells. Plasma sFLT-1 increased from 79+/-14 in NP to 172+/-34 in RUPP (p<0.05) and 107+/-9 in NP+RUPP T cells and was only 74+/-21 pg/ml in NP + αCD40L RUPP T cells.
These studies indicate that placental ischemic induced T cell communication with B cells is one important mechanism of PE that leads to much of the pathophysiology of the disease.
Abstract 048: The Sphingosine-1-phosphate Receptor S1P1 Regulates Kidney Vascular Development and Heart Development
Authors: Hu Yan , Sequeira Lopez Maria Luisa 
Published: March 2022

Sphingosine 1-phosphate receptor 1 (S1P1) is one of the five G-protein coupled receptors activated by Sphingosine 1-phosphate, which is a bioactive sphingolipid metabolite crucial in many biological processes, including vascular development. It has been reported that deletion of S1P1 in mice results in embryonic lethality at E12.5-14.5 due to failure of migration and/or differentiation of vascular smooth muscle cells (vSMCs) and pericytes and that S1P1 functions autonomously in endothelial cells (ECs) as an inhibitor of angiogenesis. However, due to the early lethality of the aforementioned S1P1-/- mice, the role of S1P1 in kidney vascular development has not been determined. This study was designed to test the hypothesis that the S1P1 is crucial for the differentiation, assembly and maturation of the renal vasculature.
Using EC-SCL-CreERT mice that specifically express tamoxifen inducible Cre in EC progenitors crossed with S1P1fl/fl and Rosa-LacZ reporter mice, we generated mice with timed deletion of S1P1 in ECs during kidney development (EC-S1P1KO). The knockout embryos with tamoxifen treatment develop severe edema, hemorrhages (36%, 8/22) or die (50%, 11/22) at E14.5 to E16.5, which may be caused by heart failure. Using histological methods including X-gal staining and immunohistochemistry for markers of ECs and mural cells, we found that the EC-S1P1KO embryos develop abnormal glomerular capillaries, dilated renal arteries and veins, disrupted vSMC coating of renal arteries and arterioles and absence of lymphatic endothelium. Further, thinner inter-ventricular septum, thinner ventricular myocardial compact layer and dilated myocardial capillaries are found in EC-S1P1KO embryos. Cross-transplantation studies of E12.5 kidneys from EC-S1P1KO and control mice under the kidney capsule of WT adult mice revealed that the kidney vascular abnormalities are intrinsic, due to the lack of S1P1 in renal EC precursors, and not secondary to the extra-renal anomalies.
These studies suggest that endothelial S1P1 is an essential regulator of both kidney vascular development and heart development.