bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 068: Ghrelin Restores Nitric Oxide Availability In The Forearm Microcirculation Of Essential Hypertensive Patients
Authors: Virdis Agostino , Duranti Emiliano , Dell'Agnello Umberto , Lorenzini Gianni , Taddei Stefano
Published: July 2021

Objective:
Essential hypertensive patients (EH) are characterized by endothelial dysfunction caused by a reduced nitric oxide (NO) availability due to reactive oxygen species excess and low-grade inflammatory condition. Ghrelin is a recently identified growth hormone-releasing peptide, with recognized cardiovascular actions. Possible effects on endothelial dysfunction have been never investigated in EH. In this study we evaluated whether exogenous ghrelin can improve endothelial dysfunction in the forearm microcirculation of untreated mild-moderate EH.

Methods:
In 9 EH (51.8±8.1 yrs) and 9 normotensive subjects (NS, 50.5±3.5 yrs), we studied the forearm blood flow (FBF, strain-gauge plethysmography) response to intrabrachial acetylcholine (ACh, 0.15-15 mg/100 ml/min) with and without NO synthase blockade by L-NMMA (100 μg/100 ml/min), or the antioxidant vitamin (Vit) C (8 mg/100 ml/min). The protocol was repeated under exogenous ghrelin intra-arterial infusion (200 ng/min, 30’ pre-infusion).

Results:
In NS, maximal vasodilation (VD) to ACh (480±20%) was inhibited by L-NMMA (292±22, -39±7%; P<0.001) and unchanged by Vit C (482±34%). Ghrelin failed to modify these vascular responses. In EH, VD to ACh was blunted vs NS (337±45%; P<0.001) and resistant to L-NMMA (313±32, -7±3%). Vit C increased the response to ACh (509±57%; P<0.01 vs ACh alone) and restored the inhibiting effect of L-NMMA (332±42, -34±8%; P<0.001). Ghrelin, while not modifying the basal FBF, it increased (P<0.001) the VD to ACh (448±55%) and restored the inhibitory effect of L-NMMA on ACh (355±43, -20±6%; P<0.001). Vit C only slightly improved VD to ACh under ghrelin infusion (486±45%). In EH ghrelin significantly (P<0.05) decreased plasma venous malodialdehyde (from 6.9±1.5 to 5.2±1.0 μmol/L), lipoperoxides (from 9.1±1.9 to 6.6±2.3 μmol/L) and IL-6 (from 11.1±0.6 to 9.3±1.0 pg/mL) and increased plasma antioxidant capacity (from 407±109 to 630±97 mmol/L). Response to sodium nitroprusside was similar between EH and NS and not affected by ghrelin.

Conclusions:
Exogenous ghrelin is able to ameliorate endothelial dysfunction by restoring NO availability in the forearm microcirculation of EH, an effect probably determined by antioxidant and/or anti-inflammatory activities.
Abstract 069: Isoketals in Monocyte-Derived Dendritic Cells Activate T Cells and Promote Hypertension
Authors: Loperena Roxana , Kirabo Annet , Davies Sean S , Roberts LJ , Harrison David G
Published: July 2021

Fatty acid oxidation leads to formation of highly reactive γ-ketoaldehydes termed isoketals that adduct to protein lysines. We have shown that hypertension causes isoketals to accumulate in dendritic cells (DCs) and activate T cells. Human monocytes traversing the endothelium differentiate into DCs expressing CD83, MHC-II, and CD11c. We hypothesized that oxidative stress catalyzes the formation of isoketals which alters monocyte and DC function to promote monocyte transformation to DCs and T cell activation. Thus, we co-cultured human monocytes with aortic endothelial cells exposed to either a hypertensive 10% stretch or a normotensive 5% stretch for 48 hours using the Uniflex® culture system. We used flow cytometry to detect human DC markers (CD14-/CD83+) in monocytes exposed to stretch. We detected conversion of CD14-/CD83+ from CD14+ human monocytes and found they contained increased levels of isoketal-ligated proteins in the 10% compared to 5% stretch (77.47 ± 7.3 vs. 12.14 ± 3.5). We also found a 1.8 fold increase of the CD86 activation marker compared to controls. Exposure of murine monocyte-derived DCs to tert-butyl hydroperoxide (t-BHP) led to a 3-fold increase in isoketals and a 2-fold increase in CD86 in the CD11b+/CD11c+ population compared to controls. Isoketal formation in DCs exposed to t-BHP was scavenged with pre-treatment of 2-hydroxybenzylamine (2-HOBA) in vitro. DCs treated with t-BHP were co-cultured with T cells for 7 days. This promoted T cell survival and proliferation of CD8+ and CD4+ T cells compared to untreated controls; this effect was attenuated with pre-treatment of 2-HOBA. Moreover, adoptive transfer of t-BHP treated DCs to normal mice elevated the hypertensive response to a generally subpressor dose of angiotensin-II (128 ± 0.80 vs. 114 ± 0.48 in controls). We conclude that oxidant stress and isoketal formation in murine DCs promote T cell activation and hypertension. We hypothesize that exposure to hypertensive stretch in human endothelial cells transfers an oxidant signal to monocytes and promotes their transformation to DCs, which mature and promote an immune response. These findings provide a mechanism as to how T cells are activated in hypertension and provide insight into the inflammatory nature of this disease.
Abstract 073: The Absence Of Renal Angiotensin-converting Enzyme Prevents Salt Sensitive Hypertension
Authors: Giani Jorge F , Janjulia Tea , Han Jiyang , Rodriguez-Iturbe Bernardo , Bernstein Kenneth E , McDonough Alicia A , Gonzalez-Villalobos Romer A
Published: July 2021

We previously showed that the renal ACE/angiotensin (Ang) II pathway is a master switch for sodium transport along the nephron. Further, we hypothesized that this pathway plays a key role in the development of salt sensitive (SS) hypertension. To test this, wild-type mice (WT) and mice lacking renal ACE (ACE 10/10) were exposed to post-L-NAME hypertension: L-NAME, 4 weeks → Washout, 1 week → high salt diet (HS, NaCl 4%), 3 weeks. In WT mice, L-NAME increased systolic blood pressure (SBP) from 110±2 to 135±3 mmHg (p<0.01; n=12). SBP then returned to baseline during the washout and increased again in response to the HS diet (131±3 mmHg; p<0.01). Thus, WT mice, previously salt resistant, become salt sensitive. Remarkably, ACE 10/10 mice, despite being equally hypertensive during the L-NAME phase, DID NOT develop salt sensitivity. At the end of HS phase, the ACE 10/10 SBP was 106±5 mmHg (NS). While baseline renal Ang II levels were similar (by IHC), HS diet induced a greater renal Ang II accumulation in WT vs. ACE 10/10 (26±2 vs. 13±2 % of positive staining/area; p<0.01). After 24 hr of sodium load, WT mice had a positive sodium balance (1841±360 μmol/day/100g b.w) that was later corrected at the expense of hypertension. In contrast, ACE 10/10 mice showed an enhanced natriuretic response that resulted in less sodium accumulation (604±300 μmol/day/100g b.w; p<0.01 vs. WT) and the absence of hypertension. In response to the salt load, glomerular filtration rate (GFR) was higher in ACE 10/10 compared to WT (1595±49 vs. 1451±49 μl/min/100g b.w after 24 h; p<0.01). Eventually, GFR returned to baseline values in both groups (WT: 1408±37; ACE10/10: 1393±42 μl/min/100g b.w.; NS). The analysis of key sodium transporters revealed that the natriuretic response of ACE 10/10 was associated with lower phosphorylated (p) NKCC2, pNCC and less ENaC α subunit abundance (33%, 31% and 25% reduction vs. WT, respectively; p<0.01). Furthermore, after 7 days of HS diet, ENaCα remained lower in ACE 10/10 vs. WT (31% reduction; p<0.01). These data indicate that renal ACE plays an important modulatory role in the GFR and sodium transporter responses that induce salt sensitivity.
Abstract 076: Urinary Exosomal Epithelial Sodium Channel And Sodium-chloride Cotransporter Measurement In Humans
Authors: MUSTAPHA TAOPHEEQ A , NWAZUE VICTOR , SCHEY KEVIN , SATISH RAJ , LUTHER JAMES M
Published: July 2021

Sodium reabsorption in the distal nephron is tightly regulated in part by epithelial sodium channel (ENaC) and sodium chloride cotransporter (NCC), although non-invasive measure of these proteins in humans has not previously been feasible. We recently analyzed the urinary exosomal proteome and identified candidate targets for quantification of ENaC and NCC using targeted mass spectrometry.

To test the hypothesis that urinary exosomal ENaC and NCC are altered during renin-angiotensin-aldosterone system activation, we activated the endogenous RAAS using a low sodium diet (LS) in two separate studies. We provided 8 subjects LS diet (10mmol/day for 7days) to assess urinary protein excretion at 7 days (study 1) and longitudinally over the course of 1 week (study 2). Daily 24-hour urine was collected to monitor sodium balance, and spot urine samples were obtained each morning on days 0, 2, 4, and 6 of LS diet. Urinary exosomal ENaC-α, ENaC-γ, and NCC peptides were analyzed using targeted multiple-reaction-monitoring analysis quantified with stable-isotope peptide standards, and results were normalized to urine creatinine concentration.

In study 1, urinary ENaCγ increased after 8 days of LS diet (Figure A). In study 2, urinary exosomal ENaCγ (Figure B) and NCC peptides (Figure C) increased in a time-dependent manner during LS diet. These measures of urinary sodium channel expression may provide further insight into distal sodium reabsorption in human hypertension.
Abstract 080: Selective Knockout of the Intracellular Isoform of Renin in the Brain Contributes to Metabolic and Cardiovascular Control
Authors: Shinohara Keisuke , Folchert Matthew D , Weidemann Benjamin J , Liu Xuebo , Morgan Donald A , Rahmouni Kamal , Grobe Justin L , Sigmund Curt D
Published: July 2021

Renin gene expression is regulated by two distinct promoter-first exon combinations that target renin for either secretion (exon 1a initiating secreted renin, sREN) or for cytoplasmic retention (exon 1b initiating intracellular renin, icREN). The icREN isoform is expressed predominantly in the brain and its expression is downregulated whereas sREN expression is upregulated by deoxycorticosterone (DOCA)-salt suggesting each isoform may be differentially regulated. We generated mice that lack icREN, but preserve sREN, by flanking exon-1b and its surrounding sequences (including the promoter) with loxP sites and breeding successively with mice expressing flipase and cre-recombinase. Real time quantitative RT-PCR analysis revealed a loss of icREN mRNA in the brain, but a preservation of sREN mRNA in the kidney. Total body weight was normal in male icREN-KO mice (26.3±0.7 g, n=8, 12 wk) compared to controls (25.3±0.7 g, n=12, 12 wk), but total (2.14±0.09 g vs 2.46±0.11 g) and relative (9.0±0.5 % vs 10.7±0.6%) fat mass as measured by NMR were reduced in 8-week icREN-KO mice (n=14-16, P<0.05). Whereas food intake was normal, there was an increase in resting metabolic rate as measured by respirometry in icREN-KO (0.1484±0.0036 kcal/hr, n=41, P=0.02 by ANCOVA) vs controls (0.1389±0.0038 kcal/hr, n=38). There was also a trend toward decreased digestive efficiency (81.1±1.0%, n=6, P=0.06) vs controls (84.9±1.2%, n=3) at 22 wk of age. Systolic blood pressure was decreased in icREN-KO mice (113±1 mmHg, n=12, 17.0±1.2 wk vs 120±3 mmHg, n=5, 15.9±1.9 wk, P<0.05). We previously reported a functional link between the synthesis and action of angiotensin-II and the action of leptin. We tested if this was impaired in icREN-KO mice by measuring the renal sympathetic nerve activity (RSNA) response to acute intracerebroventricular (ICV) administration of leptin. The decrease in arterial pressure was confirmed in this cohort of icREN-KO mice. Surprisingly, the RSNA response to leptin was greater in icREN-KO mice compared to controls (257.9±16.0%, n=3 vs 157.0±13.9%, n=4, 4 hr after leptin, P=0.01). Captopril attenuated the sympathetic response to ICV leptin. Together these data suggest that this novel icREN isoform contributes to metabolic and cardiovascular control.
Abstract 081: Reporter Mouse Strain Provides a Novel Look at Angiotensin Type-2 Receptor Distribution in Central Nervous System Cardiovascular Control Centers
Authors: de Kloet Annette D , Wang Lei , Ludin Jacob A , Hiller Helmut , Smith Justin A , Pioquinto David J , Scheuer Deborah A , Steckelings Ulrike , Krause Eric G , Sumners Colin
Published: July 2021

It is established that angiotensin-II acts at its type-1 receptor (AT1R) in the brain to increase sympathetic outflow and blood pressure, and modulate fluid balance. However, the role of the angiotensin type-2 receptor (AT2R) in the neural control of these processes has received far less attention, largely because of an inability to effectively localize these receptors at a cellular level in the brain. The present studies combine the use of a bacterial artificial chromosome transgenic AT2R-eGFP reporter mouse with recent advances in
in situ
hybridization (ISH) to circumvent this obstacle. Dual IHC/ ISH studies validated the AT2R-eGFP reporter mice by determining that eGFP and AT2R mRNA were highly co-localized within the nucleus of the solitary tract (NTS; 98.0 ± 0.18 %; 125 ± 3.6 of 127 ± 3.9 cells; n = 4). Analysis of eGFP immunoreactivity in the brain revealed localization to neurons within nuclei that regulate blood pressure and fluid balance (e.g., NTS and median preoptic nucleus [MnPO]). Additional IHC/ISH studies uncovered the phenotype of specific AT2R-eGFP cells. For example, within the NTS, AT2R-eGFP neurons primarily express glutamic acid decarboxylase-67 (GABAergic; 80 ± 2.8 %; 225 ± 12.5 of 280 ± 8.4 cells; n = 4), while only a subset express vesicular glutamate transporter-2 (glutamatergic; 18.2 ± 2.9 %; 50.8 ± 7.7 of 280 ± 8.4 cells) or AT1R (8.7 ± 1.0 %; 22 ± 2.2 of 256 ± 11.7 cells). No co-localization was observed with tyrosine hydroxylase in the NTS. Although AT2R-eGFP neurons were not observed within the paraventricular hypothalamic nucleus (PVN), eGFP was localized to efferents terminating in the PVN and to GABAergic neurons surrounding this nucleus. Retrograde neuronal tract tracing studies revealed that many eGFP-positive efferents to the PVN arise from neurons in the MnPO. Based on these neuroanatomical results, we hypothesized that activation of central AT2R would decrease blood pressure. Consistent with this hypothesis, chronic administration of the selective AT2R agonist, compound 21 (7.5 ng/h into the lateral cerebral ventricle) reduced baseline mean arterial blood pressure relative to control mice (103 ± 1.65 v. 110 ± 1.70 mmHg; n = 16; p = 0.02). These studies demonstrate that central AT2R are positioned to regulate blood pressure.
Abstract 083: Role of Endoplasmic Reticulum Stress in the Subfornical Organ (SFO) in Fluid Balance and Metabolic Effects of Brain Renin-Angiotensin System (RAS) Activation
Authors: Jo Fusakazu , Jo Hiromi , Hilzendeger Aline M , Cassell Martin D , Rutkowski D. T , Davisson Robin L , Grobe Justin L , Sigmund Curt D
Published: July 2021

Endoplasmic reticulum (ER) stress has been identified as an important contributor to neurological diseases and implicated in mediating hypothalamic inflammation and the hypertensive effects of angiotensin II (AngII). We examined the role of ER stress in the metabolic and fluid balance effects of brain AngII in two mouse models: 1) sRA transgenic mice (expressing human renin in neurons and human angiotensinogen in glia and neurons), and 2) DOCA-salt treated C57BL/6J mice. Both DOCA-salt and sRA mice exhibit hyperactivity of the brain RAS, suppression of circulating RAS, hypertension, polydipsia, and an elevated resting metabolic rate (RMR). CCAAT-enhancer-binding protein homologous protein (CHOP), a marker of chronic ER stress, was examined by immunocytochemistry in the brain of both models. CHOP immunoreactivity was evident in the SFO of sRA and DOCA-salt mice but was absent in control and CHOP-/- mice. We infused the ER stress-reducing chemical chaperone tauroursodeoxycholic acid (TUDCA, 5.28 ug/day, or aCSF vehicle) to assess if ER stress is mechanistically related to the hypertension, polydipsia, and elevated RMR observed in both models. In initial studies, ICV TUDCA significantly attenuated the polydipsia (aCSF 20.7±0.9 vs TUDCA 10.8±1.0 mL/day, n=6,2) and RMR (aCSF, 3.38±0.07 vs TUDCA 3.16±0.06 mL O2/100g/min, P<0.05 n=13,11) in the DOCA-salt model. ICV TUDCA had similar effects on the polydipsia in the sRA model (51±10% of aCSF control, P<0.05 n=3,4). In the DOCA-salt model, daily ICV injections of TUDCA (10 days, 5ug/ul) markedly reduced drinking, but polydipsia returned one day after the injections were terminated (n=14,12). Daily ICV injection of another ER stress reducer 4-phenylbutyrate (5ug/ul) also reduced drinking (P<0.05 n=5,4). To assess the functional role of CHOP, we measured RMR and water intake in CHOP-/- mice. Interestingly, CHOP-/- mice exhibited increased baseline RMR (CHOP-/- 0.161±0.010 vs C57 0.140±0.005 kcal/hr, P<0.05 n=10,9). The increase in water intake in response to DOCA-salt was blunted (32.7±0.5 vs 22.8±1.1 ml/day, P<0.05, n=4,5) in CHOP-/- mice. Together these data mechanistically implicate ER stress in the fluid and metabolic responses to increased brain RAS activity and suggest CHOP may play a functional role.
Abstract 086: Adenosine A3 Receptors Regulate Oxidative Stress And Inflammatory Responses In A Model Of Hypertension And Renal Disease
Authors: Yang Ting , Hezel Micheal , Peleli Maria , Zhang Xingmei , Terrando Niccolo , Fredholm Bertil , Carlström Mattias
Published: July 2021

BACKGROUND:
Adenosine A1 and A2 receptors are known to regulate renal autoregulation and blood pressure, but the role of A3 signaling is unknown. We previously demonstrated early life reduction in nephron number (UNX) combined with high salt diet (HS) induced renal oxidative stress and hypertension in rats. This study aimed at investigating the role of A3 receptor in modulating renal and cardiovascular function in this disease model.

METHODS:
Wild-type (WT) and A3 knockout (A3KO) mice underwent UNX or sham-operation at 3-week-age followed by HS treatment. Blood pressure and renal function were measured in conscious aged mice and renal oxidative stress as well as inflammatory properties was characterized.

RESULTS:
WT but not A3KO with UNX+HS developed hypertension and cardiac hypertrophy (WT: 4.5±0.1 vs 4.0±0.1; A3KO: 4.7±0.2 vs 4.6±0.1 mg/gBW), characterized by impaired renal plasma flow (RPF), glomerular hyperfiltration and increased renal NADPH oxidase activity (Fig. 1A-C). UNX+HS increased plasma IL-6 (36.1±5.7 vs 16.3±2.4 pg/ml) and IL-10 (37.8±3.9 vs 25.4±2.4 pg/ml) levels in WT, but not in A3KO (39.7±3.9 vs 38.5±5.9 pg/ml and 46.5±3.7 vs 43.9±6.0 pg/ml respectively). However, A3KO displayed higher baseline cytokine levels. Furthermore, bone marrow-derived macrophages from A3KO mice expressed higher M1 (PDL-1 and CD86) and M2 (PDL2 and CD206) markers under LPS stimulation compared with WT. These suggested an enhanced innate immune response in A3KO.

CONCLUSION:
UNX followed by HS intake lead to renal and cardiovascular dysfunction, which importantly depends on A3 receptor-mediated regulation of oxidative stress and inflammation.
Abstract 088: Angiotensin Converting Enzyme 2 (ACE2) in the Kidney: Soluble ACE2 Restores Blood Pressure Regulation
Authors: Gurley Susan , Ortiz Melo David , Ortiz Natalie M , Prescott Jessica , Xu Katherine , Griffiths Robert , Burns Kevin
Published: July 2021

Angiotensin converting enzyme 2 (ACE2) is a carboxypeptidase that metabolizes angiotensin II (AngII) to Ang(1-7). ACE2 can be cleaved by metalloproteinases to generate soluble ACE2 (sACE2), which remains catalytically active. The objective of this study is to determine if sACE2, acting in the kidney, could modulate the BP response to AngII infusion.Data from our kidney cross-transplant study using male (129/SvEv x C57BL/6)F
1
ACE2 KO and WT mice suggest that sACE2 can reach the urinary compartment even when not expressed in kidney cells. For these experiments, we generated 4 experimental groups: 1) WT, 2) Kidney ACE2KO, 3) Systemic ACE2KO, and 4) Total ACE2KO. Mean arterial pressures (MAP) were measured with radiotelemetry. AngII was infused chronically at 1ug/kg/min. Enzymatic activity was determined using an

ACE2-specific quenched fluorescent substrate assay. ACE2 protein in the urine was confirmed by western blots using a specific N-terminus antibody.

Total ACE2KO mice had an enhanced hypertensive response to AngII compared to WT (152±6 vs.144±7 mmHg;
p
=0.03). The presence of ACE2 in either kidney or systemic tissues restored MAPs to levels similar to WT (kidney ACE2KO 140±7mmHg, systemic ACE2 KO 140±5 mmHg, p<0.01 vs. total ACE2KO). However, Kidney ACE2KO mice had similar urinary ACE2 activity as systemic ACE2KO (76±32 and 108±52 RFU/ul/hr, respectively, p=NS). We confirmed this was ACE2 with immunoblotting of urine samples. As expected, we detected a 92kDa band in WT and systemic KO mice, which represents full length ACE2. A smaller band (70kDa) was present in the WT, systemic KO,
and
kidney KO groups, and absent in total KO mice. This likely represents sACE2. Moreover, the presence of the smaller band correlated with urine ACE2 activity and was associated with attenuated BPs.

Our studies demonstrate that the exaggerated hypertensive

phenotype seen in total ACE2 deficiency can be rescued when ACE2 is restored either in the kidney or in systemic tissues. Since the Kidney KO group demonstrated ACE2 activity and protein in the urine, we suggest that soluble ACE2 can be filtered in hypertensive renal injury and may provide an additional source of balance against upregulation of the RAS within the kidney, with the overall goal of regulating BP.
Abstract 097: EGF Receptor And ER Stress Mediate End-organ Damage But Not Hypertension Induced By Angiotensin II In Mice
Authors: Elliott Katherine J , Tsuji Toshiyuki , Obama Takashi , Takayanagi Takehiko , Forrester Steven , Park Joon , Eguchi Satoru
Published: July 2021

In cultured vascular smooth muscle cells (VSMC), a metalloprotease ADAM17-dependent EGF receptor (EGFR) activation mediates ERK1/2 activation and subsequent hypertrophy induced by angiotensin II (Ang II). Both ER and oxidative stress are implicated in hypertensive end-organ damage. We hypothesized that pharmacological inhibition of EGFR may prevent end-organ damage but not hypertension in mice infused with Ang II via suppression of ER/oxidative stress. To test this hypothesis, we have evaluated Ang II-induced end-organ damage as well as hypertension in C57Bl/6 mice with or without an EGFR inhibitor erlotinib (20mg/kg/day ip) or ER stress inhibitor 4-phenyl butyric acid (PBA: 1g/kg/day in drinking water). Upon Ang II infusion (1000 ng/kg/min) for 2 weeks, control mice showed phenotypes of cardiac hypertrophy including increased HW/BW ratio (mg/g: 7.9±0.7 vs 5.7±0.6 p<0.01 n=8) and increased LVPWd assessed by cardiac echo (mm: 0.98±0.14 vs 0.69±0.05, p<0.05 n=8) compared with saline infusion. Histological assessments demonstrated medial hypertrophy and perivascular fibrosis of coronary arteries with Ang II infusion. In contrast, cardiac hypertrophy and vascular remodeling were attenuated in mice with Ang II plus erlotinib or PBA; HW/BW ratio (6.8±0.6 or 6.2±0.6 n=8), and cardiac echo (LVPWd: 0.65±0.07 or 0.80±0.07 n=8) compared with saline infusion. Renal fibrosis observed with Ang II infusion was also attenuated in mice with Ang II plus erotinib or PBA. However, Ang II induced similar levels of hypertension in non-treated, erlotinib-treated or PBA-treated mice assessed by telemetry (MAP mmHg: 144±9 vs 149±20 or 139±4). Ang II infusion in mice enhanced ADAM17 and phospho-Tyr EGFR staining in vasculatures of heart and kidney, whereas mice with Ang II plus erlotinib or PBA had diminished phospho-Tyr EGFR staining and no ADAM17 staining in the vasculatures. In addition, IHC analyses revealed less oxidative stress and less ER stress in heart and kidney of Ang II-infused mice with erlotinib or PBA. These data suggest that EGFR activation and subsequent ER stress enhancement are critical for end organ damage via induction of ADAM17 and oxidative stress and is independent from blood pressure regulation.
Abstract 101: Chemerin, A Novel Adipokine, Regulates Human Vascular Cell Function Through Redox-sensitive Processes Involving NADPH Oxidase 1/4 And Endothelial Nitric Oxide Sinthase.
Authors: Neves Karla B , Lopes Rheure A , Nguyen Aurelie D , Lobato Nubia S , Oliveira Ana Maria , Montezano Augusto C , Tostes Rita C , Touyz Rhian M
Published: July 2021

Adipose tissue produces many adipokines, including chemerin, a chemoattractant for inflammatory cells that mediates its effects through chemokine-like receptor 1 (CMKLR1). Chemerin has been implicated in endothelial dysfunction and vascular inflammation, hallmarks of vascular injury in hypertension. These pathological processes are induced by reactive oxygen species (ROS). How chemerin regulates ROS is unknown. We postulated that chemerin, through redox-sensitive mechanisms, influences signalling in vascular cells. Human vascular smooth muscle cells (HVSMCs) and microvascular endothelial cells (HMECs) were studied. HVSMCs and HMECs were stimulated with recombinant chemerin (50ng/ml). eNOS and MAPK activation were assessed by immmunoblotting, ROS generation by chemiluminescence and nitric oxide (NO) levels by DAF-FM fluorescence. mRNA expression of Nox 1, Nox 4 and inflammatory markers was assessed by real time PCR. Chemerin stimulated ROS generation in HVSMCs (1 h - 21% increase; 8 h - 41% increase) and HMECs(1 h - 54% increase; 24 h - 76% increase, p<0.05 vs vehicle), effects blocked by Nox1/4 inhibitor (GKT137831) and Nox1 inhibitor (ML171). Chemerin also increased phosphorylation of ERK1/2 (43% and 62%), p38MAPK (18% and 25%) and JNK (59% and 73%) in HVSMCs and HMECs respectively. eNOS phosphorylation (Ser1177, activation site) was decreased by chemerin in HMECs (45% decrease), while phosphorylation at nhibitory site, Thr495, was increased (72%). In parallel, chemerin decreased NO levels in HMECs by 33%. Finally, chemerin increased IL-6 (290%), MCP-1 (64%), VCAM-1 (210%) and ICAM-1 (130%) mRNA levels in HVSMCs. Nox 1 (49%) and Nox 4 (67%) mRNA levels were increased in HVSMCs stimulated with chemerin. In conclusion in human vascular cells, chemerin stimulates ROS generation and reduces NO formation, through Nox1/4 activation and eNOS inhibition respectively. These processes were associated with increased redox-sensitive MAPK signaling and inflammation. Our study identifies chemerin as a new vasoactive factor, that plays an important role in vascular injury and dysfunction, which may be particularly important in the context of increased adipocyte-derived chemerin, such as in obesity, metabolic syndrome and hypertension.
Abstract 200:

Low Dose Spironolactone Treatment Prevents Cardiac Diastolic Dysfunction, Oxidant Stress and Fibrosis in a Female Model of Obesity in Concert with a Modulation of Inflammation
Authors: Bostick Brian , Habibi Javad , McKarns Susan C , Ma Lixin , Rehmer Nathan T , Bender Shawn , Garro Mona , Meuth Alex , Nistala Ravi , Jia Guanghong , Sowers James R
Published: July 2021

Obesity is a global epidemic with 1.5 billion overweight or obese people worldwide. The association of obesity and a high-fat/high-fructose Western diet (WD) markedly increases cardiovascular disease (CVD) particularly diastolic dysfunction in women. Few treatments exist for diastolic heart disease but, recent work implicates the mineralocorticoid receptor (MR) in inflammation, fibrosis and oxidant stress. We thus hypothesized that low-dose spironolactone (LDSp) could prevent diastolic dysfunction by reducing fibrosis, oxidant stress and inflammation. To test this hypothesis, we developed a female model of obesity induced diastolic dysfunction and examined preventative treatment with LDSp.

Four week-old C57BL6/J female mice were fed a WD with/without 1.0mg/kg/day of Sp (n=7 for each group). This dose of LDSp exerts no effect on blood pressure. After 16 weeks, we conducted detailed phenotypic analysis and assessed diastolic function by cardiac MRI. Immunohistochemistry was also done for cardiac oxidant stress, fibrosis, collagen content and insulin receptor (IRS-1) expression. Flow cytometry of heart tissue was utilized to examine inflammatory mechanisms.

We found WD feeding caused diastolic dysfunction that was prevented by LDSp (LV diastolic relaxation time 33.4 ± 1.2ms for WD, 20.6 ± 1.5ms for control and 24.3 ± 0.9ms for WD+LDSp, p <0.01). 3-nitrotyrosine staining showed significant cardiac oxidant stress with WD feeding that was ameliorated by LDSp. Cardiac fibrosis and the collagen 1:3 ratio were markedly increased with WD but prevented by LDSp. IRS-1 expression and phosphorylation were markedly reduced by WD feeding but not improved by LDSp. Flow cytometry showed evidence towards reduced M2 macrophage polarization with lower CD11b/CD301 double positive cells in WD fed hearts with trend toward improvement with LDSp.

These findings suggest WD induces cardiac diastolic dysfunction in females by increasing oxidant stress and fibrosis potentially mediated by deficiency in anti-inflammatory M2 macrophages. MR antagonism with spironolactone may prevent the decrease in M2 macrophage polarization reducing oxidant stress and fibrosis. This work supports a novel mechanism for spironolactone in treatment of WD induced heart disease.
Abstract 205: Potential Cardiovascular Risk Reduction in High-Risk Uninsured Patients: An Analysis of the Heart Improvement Project
Authors: Thomas Joshua R , Chandrasekharan Gopal , Zhao Daniel , Fox Mark D
Published: July 2021

Background:
The Heart Improvement Project (HIP) is a cardiovascular (CV) risk reduction clinic targeting high-risk, uninsured patients. Using coronary heart disease prediction models from the Framingham Heart Study, it is possible to project an individual’s 10-year CV disease risk based on status of the following risk factors: sex, age, LDL-Cholesterol (LDL-C), HDL-Cholesterol (HDL-C), blood pressure (BP), diabetes, and smoking status.

Objective:
To project the reduction in CV risk associated with two levels of intervention: first-line therapy (designed to decrease the systolic BP by 15 mm Hg and LDL-C by 30%) and maximal therapy to optimally control all modifiable risk factors.

Methods:
Data on 251 patients (52% male, mean age 51 yrs; 48% with Diabetes, 45% smokers) from HIP were analyzed. For each patient, their Framingham Risk Scores (FRS) and 10-year Coronary Heart Disease Risk (CHDR) were calculated in three different conditions: their actual risk assessment, and that projected under first-line, and maximal therapy. The three sets of FRS and CHDR were summarized using descriptive statistics. Pairwise differences between the three sets of FRS and CHDR were tested using two-sample t-test and 95% confidence intervals for the differences were computed.

Results:
Table 1 shows that the mean FRSs were 7.33, 4.90, and 3.51 (under initial, first-line, and maximal conditions, respectively). The corresponding CHDRs were 13%, 8%, and 6% respectively. All comparisons were statistically significant at the p<0.001 level.

Conclusions:
First-line interventions for BP and lipid management can yield significant improvement in CHD risk in a population of high-risk uninsured patients.
Abstract 215: Angiotensin-II-induced Cardiac Remodeling is Reduced in TNFR1-deficient Mice Despite Increased Blood Pressure
Authors: Duerrschmid Clemens , Aguirre-Amezquite Fernando , Taffet George E , Entman Mark L , Haudek Sandra B
Published: July 2021

Background:
Infusion of angiotensin-II (Ang-II) to wild-type (WT) mice results in hypertension, development of interstitial cardiac fibrosis and hypertrophy, and deterioration of myocardial function. We previously showed that after 1 week of Ang-II infusion, these effects were absent in mice deficient in tumor necrosis factor receptor 1 (TNFR1). We now investigated long-term effects of Ang-II infusion.

Methods:
WT and TNFR1-KO mice were infused with Ang-II for 6 weeks. Systolic blood pressure (SBP) was measured by tail-cuff plethysmography; cardiac function by 2D-echocardiography and Doppler ultrasound. Hearts were analyzed for collagen deposition (histology) and expression of fibrosis- and hypertrophy- related genes (quantitative PCR).

Results:
In WT mice, SBP increased within 7 days and remained elevated at 6 weeks (152±4 mmHg); cardiac fibrosis developed after 1 week and persisted at 6 weeks (6.2±1.1% collagen area). By contrast, in TNFR1-KO mice, SBP at 7 days was low, but increased by 6 weeks (144±4 mmHg), whereas cardiac fibrosis was absent at 1 week and did not significantly increase by 6 weeks (2.5±0.5%). In support of these data, collagen I and collagen III mRNA expression at 6 weeks were upregulated in WT (2.9±0.6 and 4.1±0.8 -fold over sham), but not in TNFR1-KO hearts (1.3±0.1 and 1.8±0.2). In both mouse groups, cardiac hypertrophy and cardiac dysfunction developed over time, however, these changes were less prominent in TNFR1-KO mice: at 6 weeks, the heart-weight to body-weight ratio in WT was 6.7±0.4, in TNFR1-KO mice 5.5±0.2; the changes in anterior and posterior wall thicknesses in WT were 44±12% and 32±15%, in TNFR1-KO mice 19±8% and 17±10%; the change in ejection fraction in WT was -67±12%, in TNFR1-KO mice -39±5%; and the change in Tei-index (myocardial performance) in WT was 18±9%, in TNFR1-KO -1±7%. Also, hypertrophy-related atrial natriuretic peptide (ANP) and beta-myosin heavy chain (b-MHC) mRNA were upregulated in WT (4.3±0.9 and 4.3±0.6 -fold over sham), but less in TNFR1-KO hearts (2.6±0.5 and 2.4±0.5).

Conclusion:
Despite a significant increase in blood pressure over 6 weeks of Ang-II infusion, TNFR1-KO mice developed less cardiac fibrosis and hypertrophy and had better cardiac function compared to WT mice.
Abstract 222: Upregulation Of Cyp4a1 Gene Expression Restores The Impaired Myogenic Response And Autoregulation Of Cerebral Blood Flow In Dahl Salt Sensitive Rats
Authors: Fan Fan , Pabbidi Mallikarjuna R , Geurts Aron M , Jacob Howard , Roman Richard J
Published: July 2021

We have reported that a reduction in the expression of CYP4A and the production of 20-HETE in the renal outer medulla contributes to development of hypertension in Dahl salt sensitive (SS) rats. The present study examined whether 20-HETE production is also reduced in the vasculature and if a deficiency in the formation of 20-HETE in the vasculature alters vascular tone and promotes end organ damage. The production of 20-HETE, the myogenic response of middle cerebral arteries (MCA) and autoregulation of cerebral blow flow (CBF) was compared in SS, CYP4A1 transgenic SS rats and SS.5BN consomic rats in which chromosome 5 from Brown Norway (BN) was transferred into the SS genetic background. 20-HETE production was 6-fold higher in cerebral arteries obtained from CYP4A1 transgenic (n=25) and SS.5BN (n=4)rats than in SS (n=17) rats 0.77 ± 0.08 versus 0.12 ± 0.03 pmol/mg/min). The luminal diameter of MCA decreased to 70 ± 3 % in CYP4A1 transgenic rats and to 65 ± 6 in SS.5BN when perfusion pressure was increased from 40 to 140 mmHg, whereas it remained unaltered in SS rats. Administration of the inhibitor of the synthesis of 20-HETE, HET0016 (10 uM), abolished the myogenic response in MCA of CYP4A1 transgenic and SS.5BN rats but had no effect in SS rats. CBF was poorly autoregulated and increased by 49 ± 6% in SS rats as MAP was increased by 60% from 100 to 160 mmHg. In contrast, CBF was only increased by 26 ± 5% and in SS.5BN rats and by 19 ± 2% in CYP4A1 transgenic rats when MAP was increased in the same range. These results indicate that a genetic deficiency of 20-HETE contributes to an impaired myogenic response in autoregulation of CBF in SS rats which may contribute to vascular remodeling, stroke and dementia following the onset of hypertension.
Abstract 228: Apolipoprotein E (ApoE) Genetic Polymorphism Modulation Of Endothelial Dysfunction In Hypertensive Patients Is Determined By Renal Function.
Authors: Marrocos Mauro S , Teixeira Andrei A , Quinto Beata M , Rodrigues Cassio Jose de Oliveira Rodrigues J , Dalboni Maria A , Manfredi Sylvia , Canziani Maria Eugênia Canziani E , Batista Marcelo C
Published: July 2021

INTRODUCTION:
ADMA, an endogenous inhibitor of nitric oxide synthase, is an independent marker of progression of renal dysfunction, vascular complications and death. Overexpression of the ADMA degrading enzyme dimethylarginine dimethylaminohydrolase 1 (DDAH1) ameliorates atherosclerosis in genetic ApoE deficient mice. Once ApoE polymorphism is a key player on lipid metabolism, it is possible that it brings on individual susceptibility to endothelial dysfunction.

OBJECTIVE:
The aim of this study was to assess the effect of the ApoE genetic polymorphism on ADMA levels in a cohort of hypertensive patients stratified by renal function.

PATIENTS AND METHODS:
Patients were stratified into 3 groups according to GFR using the CKD-EPI: I > 60ml/min, II ≤ 60 ml/min and > 15 ml/min, and III ≤ 15 ml/min. HPLC was used for the measurement of ADMA. Polymorphic ApoE analysis performed by PCR amplification and allele frequency compared in each group between first and third tertile of ADMA as epidemiological, clinical and laboratorial characteristics. Statistical analysis performed using the software IBM SPSS Statistics 20.0.

RESULTS:
Mean age of the patients was 60.1± 14.1 years, 318 (50.7%) were male, mean BMI was 28.0 ± 5.3, 396 (63.2%) were Caucasians, 244 (38.9%) had DM, 316 (50.4%) had dyslipidemia, 223 (35.6%) had cardiovascular disease (CVD). Groups were composed respectively of 189, 229 and 202 patients. E2 allele was present in 67 (10.7%) patients, E3 allele in 588 (93.8%), E4 allele in 116 (18.5%). In group III, the frequency of E4 allele of ApoE was lower in the third tertile of ADMA (p = .000, Pearson Chi-Square). This link remained significant even after control for confounders like age, sex, BMI, DM, CVD, PCR and LDL (OR 0.127, CI: 0.35 - 0.468, p = .002).

CONCLUSION:
ApoE E4 allele independent association with lower levels of ADMA is pronounced among those patients with advanced stage of renal disease. This association may represent an unknown genetic pathway that modulates ADMA levels on this population.
Abstract 240: Hypertension Prevalence and Management Among Adult Congenital Heart Disease Patients
Authors: Thomas Joshua R , Munoz Ricky T , Fox Mark D , Yetman Angela T
Published: July 2021

Background:
Cardiovascular risk factors (CRFs) are poorly defined among the population of adults with congenital heart disease. In particular, the prevalence of pre-hypertension (pre-HTN) and hypertension (HTN) are currently unknown.

Objective:
To determine the prevalence of pre-HTN and HTN among adults with congenital heart disease; and to assess HTN control rates among different clinic types.

Methods:
A retrospective analysis of a cohort of adults with congenital heart disease (CHD) was conducted. Data regarding CHD patients' blood pressure (BP), medications, and provider specialty were analyzed.

Results:
The cohort consisted of 971 patients. The mean age was 30.4 years (SD = 10.4), with 51% male. Thirty-two percent had HTN (n = 304). There was no statistically significant difference in the prevalence of Pre-HTN and HTN by clinic type (i.e. specialty vs. non-specialty)(
p
= .225, p= .633 respectively). However, a statistically significant association exists between clinic type and HTN control rates
χ
2
(1) = 3.185,
p
= .07 (Table 1). Those receiving care from a specialty clinic are 1.6 times more likely to have controlled HTN.

Conclusions:
Pre-HTN and HTN are common among adults with congenital heart disease. These CRFs are better managed in specialty clinics devoted to this population.
Abstract 246: Role Of Cytochrome P450 4a2 In Mediating The Elevated Blood Pressure In A Rat Model Of Polycystic Ovary Syndrome
Authors: Maranon Rodrigo O , Patil Chetan N , Dalmasso Carolina , Roman Richard , Reckelhoff Jane F
Published: July 2021

Women with polycystic ovary syndrome (PCOS) often have elevated blood pressure (BP). PCOS is characterized in part by increases in androgens, and androgens can increase cytochrome P450 (CYP) 4A isoforms and 20-HETE synthesis. We have found that CYP4A2 expression is increased in renal vasculature of hyperandrogenemic female rats, a model of PCOS. In the present study we tested the hypothesis that androgen increase would not cause elevated BP in CYP4A2
-/-
rats compared with wild type SS.Bn5 rats. CYP4A2
-/-
and SS.Bn5 rats (n=6-8/grp) were treated from 4 wks of age with dihydrotestosterone pellets (DHT 7.5 mg/90 d) or placebo pellets until 14 wks, and then telemetry transmitters were implanted. After 2 wks, mean arterial pressure (MAP) was measured for 10 days. DHT increased MAP and decreased HR in SS.Bn5 compared with placebo controls (placebo: 104±2 vs. DHT: 126±6 mmHg, p<0.001). In contrast, while placebo-treated CYP4A2
-/-
rats had higher MAP than WT, DHT did not increase BP in CYP4A2
-/-
rats (Placebo: 120±1 vs. DHT: 118±1 mmHg, p=NS). These data suggest that CYP4A2 may be necessary for DHT to increase BP in our model of PCOS. However, by what mechanism(s) CYP4A2
-/-
rats have higher MAP than SS.Bn5 WT remains to be determined. Supported by NIH R01HL66072, P01HL05971 and AHA 14POST18640015.
Abstract 249: Male Mice with XX Sex Chromosome Complement Exhibit Reduced Angiotensin II-Induced Hypertension and Atherosclerosis
Authors: Alsiraj Yasir , Thatcher Sean , Cassis Lisa A
Published: July 2021

Objective:
Hypertension and atherosclerosis exhibit sexual dimorphism, partially ascribed to effects of sex hormones. However, recent studies suggest that sex chromosome complement may also contribute to sexual dimorphism of these diseases. The renin-angiotensin system has been suggested to contribute to sexual dimorphism of hypertension and atherosclerosis. However, the relative contribution of sex hormones, versus chromosomes, in the regulation of angiotensin II (AngII)-mediated responses is unknown. We hypothesized that the presence of an XX chromosome complement in male hyperlipidemic mice would reduce AngII-induced hypertension and atherosclerosis.

Methods and Results:
Male Ldlr-/- mice with an Sry mutation but with an Sry transgene on autosomes were bred to Ldlr-/- females to generate XY or XX males. Male XY or XX mice were segregated to sham-operated or orchiectomized (ORC) groups and fed a high fat diet (42% kcal from fat). Two weeks later, mice were infused with AngII (1,000 ng/kg/min) for 4 weeks. In the presence of sex hormones (sham groups), systolic blood pressure (SBP) was significantly lower in AngII-infused XX than XY males (XY, 136 ± 4; XX, 119 ± 4 mmHg, p=0.024). In XY males, ORC had no significant effect on SBP compared to sham (ORC XY, 129 ± 6 mmHg; p=0.48). However, following castration of XX males, SBP significantly increased (ORC XX 135 ± 3 mmHg; p =0.002). In XY males, ORC resulted in a significant increase in atherosclerotic lesion surface area in the aortic arch (sham, 19 ± 4; ORC, 32 ± 7%; p=0.04). In contrast, ORC had no significant effect on lesion surface area in XX males (sham, 17 ± 3; ORC, 15 ± 2%; p=0.53). Moreover, in the absence of sex hormones, lesion surface area was markedly increased in XY compared to XX males (XY, 32 ± 7; XX, 15 ± 2%; p=0.002).

Conclusions:
These results demonstrate that effects of testosterone to regulate AngII-induced hypertension and atherosclerosis are dependent on sex chromosome complement. In XY males, testosterone protects against AngII-induced atherosclerosis. In XX males, testosterone protects against AngII-induced hypertension. Moreover, there is pronounced effect of XY chromosome complement to promote AngII-induced atherosclerosis in males.
Abstract 255: Lymphocyte-Specific Adaptor Protein, LNK (SH2B3), Regulates Angiotensin-II Induced Hypertension and Renal/Vascular Inflammation
Authors: Saleh Mohamed A , McMaster William G , Funt Samuel A , Thabet Salim R , Wu Jing , Xiao Liang , Norlander Allison E , Chen Wei , Itani Hana A , Huan Tianxiao , Levy Daniel , Harrison David G , Madhur Meena S
Published: July 2021

LNK, an adaptor protein primarily expressed in hematopoietic cells and endothelial cells, is a negative regulator of cytokine signaling and cell proliferation. A single nucleotide polymorphism in LNK is associated with hypertension, but the mechanism is unknown. Our objective was to determine the effect of LNK on hypertension and inflammation using LNK
-/-
mice. In response to angiotensin II (Ang II) infusion, LNK
-/-
mice exhibit elevated systolic BP (SBP) compared to wild type C57Bl/6J mice (WT) as measured by telemetry. At baseline, kidneys from LNK
-/-
mice exhibit greater levels of total leukocyte and T lymphocyte infiltration, superoxide production, and albuminuria compared to WT mice, and these parameters are further exacerbated by AngII infusion. Aortas from LNK
-/-
mice exhibit enhanced inflammation, reduced nitric oxide production, and impaired endothelial-dependent relaxation. Bone marrow transplantation studies showed that loss of LNK in hematopoietic cells (not somatic cells) reproduced the phenotype of whole body deletion of LNK with SBP reaching 180 mmHg in response to a subpressor dose of Ang II. Splenic T cells from LNK
-/-
mice produce elevated levels of interferon-gamma (IFNγ) compared to WT mice. Furthermore, IFNγ
-/-
mice exhibit blunted hypertension in response to AngII infusion, suggesting that enhanced IFNγ production is at least partly responsible for the aggravated hypertension in LNK
-/-
mice. Data are summarized in the table (expressed as mean±SEM, n=5-10,*
P
<0.05 vs WT/Sham,
#
P
<0.05 vs WT/AngII and
†
P
<0.05 vs LNK
-/-
/Sham). In conclusion, LNK may serve as a novel therapeutic target for hypertension and its associated renovascular disorders.
Abstract 262: Mitochondrial N-formyl Peptides Lead to Sepsis-like Symptoms via Basophil Activation
Authors: Wenceslau Camilla F , McCarthy Cameron G , Goulopoulou Styliani , Szasz Theodora , Webb R. Clinton
Published: July 2021

Traumatic injury may lead to bacteria-induced systemic inflammation (sepsis). However, 15% of trauma patients present sepsis-like symptoms in the absence of infection. In these patients, it is possible that fragments from mitochondria (damage-associated molecular patterns) from traumatized tissue act to initiate an inflammatory response. Previously, we have demonstrated that infusion of mitochondrial N-formyl peptides (F-MIT) leads to exacerbated vasodilatation and hypotension via formly-preptide receptor (FPR) activation and inducible nitric oxide release. However, the cell type mediating this effect was not determined. We hypothesized that F-MIT induces sepsis-like symptoms, include hypotension, via basophil activation. Male Wistar rats (12-weeks old, n=5-8) were cannulated (femoral artery and vein) for blood pressure recordings and drug infusion. To analyze organ damage, the lungs were collected and stained with H&E. F-MIT infusion (0.02 mg/rat/
i.v.
) induced severe lung injury (edema, neuthrophil infiltration and alveolar thickening) (4-fold vs. control) and vascular leakage in aorta (+34%) and mesenteric resistance arteries (+66%). Moreover, F-MIT infusion increased body temperature (Control: 36±0.09 vs. F-MIT: 37±0.28 °C, *p<0.05) and reduced bleeding time (Control: 89±8 vs. F-MIT: 33±11 seconds, *p<0.05). Since F-MIT induced vascular leakage, rats were treated with histamine 2 receptor antagonist, cimetidine (50 mg/kg/
i.v.
) prior F-MIT infusion. As expected, F-MIT (0.02 mg/rat) induced hypotension [Control: 82±3 vs. F-MIT: 60±4 mmHg, *p<0.05]. However, cimetidine treatment abolished this response (Control: 75±1 vs. F-MIT infusion: 70±4 mmHg). Interesting, F-MIT had no effect on blood pressure in animals treated with anti-asialo GM1 antiserum (0.2 ml/
i.p
.) for depletion of basophils. Neither neutrophil nor mast cell depletion blocked F-MIT-induced hypotension (p>0.05). These data suggest that F-MIT, via basophil activation, leads to histamine release, vascular leakage and hypotension, associated to organ damage, fever, blood clotting. Thus, F-MIT may be the link between SIRS and cardiovascular collapse in sterile trauma patients.