bluuebunny/crossref_metadata_2025_split huggingface dataset:
The characterization of SPX‐101, a new peptide promoter of ENaC internalization, revealed basic mechanistic understanding of ENaC internalization and downstream signaling events.
Authors: Walker Matthew Paul , Sesma Juliana , Wu Bryant , Stuhlmiller Timothy , Scott David 
Published: November 2023

Cystic Fibrosis (CF) is a life‐altering disease that affects over 70,000 people worldwide. CF is defined by mutations in Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) causing an improperly functioning protein or loss of protein expression. Loss of CFTR activity causes an imbalance in ion levels leading to defects in mucus clearance and acidification of the lung. A major effect of lung acidification is the loss SPUNC1 activity. SPLUNC1 is secreted to the airway surface liquid and regulates ENaC through direct binding to the extracellular region of the channels. This interaction is lost at low pH, resulting in ENaC accumulation on the membrane and dehydration of the lung. Restoring SPLUNC1 regulation of ENaC is an attractive target for CF therapy, as it will increase lung hydration allowing for increased mucus clearance and normalization of normal lung function. SPX‐101 is a SPLUNC1‐derived peptide that internalizes ENaC regardless of pH. SPX‐101 can be nebulized directly into the lung allowing for efficient drug delivery into the target organ while achieving low systemic circulation. SPX‐101 has shown significant efficacy in CF animal models and safety in pre‐clinical toxicology and phase 1 clinical trials.Mechanistically, SPX‐101 internalized ENaC in a dose dependent manner in CF‐derived human bronchial epithelial cell (HBEC), decreased amiloride sensitive current, and increased airway surface liquid. SPX‐101 induced ENaC ubiquitination causing a robust and prolonged decrease in the membrane levels of ENaC alpha, beta, and gamma. ENaC internalization was achieved through activation of a pro‐internalization kinase axis followed by a compensatory increase in signaling mediators responsible for regulating ENaC stability. This internalization and downstream signaling activation is markedly different than ENaC channel blockers such as amiloride or derivatives of the parent molecule. Amiloride and its derivatives do not induce ENaC internalization or regulate downstream kinases, suggesting that these downstream signaling events are caused by internalization and/or SPX‐101 induced changes in ENaC structure rather than changes in sodium secretion. Together this work provides the molecular basis for the activity of the first in class peptide promoter of ENaC internalization, SPX101, and provided valuable insight into the mechanisms by which ENaC proteins are regulated on the cell surface.
Antibody‐antibiotic conjugates: a new platform for treatment of serious bacterial infections
Authors: Brown Eric J 
Published: November 2023

Staphylococcus aureus is considered to be an extracellular pathogen. However, survival of S. aureus within host cells may provide a reservoir relatively protected from antibiotics, thus enabling long‐term colonization of the host and explaining clinical failures and relapses after antibiotic therapy. We have shown that intracellular S. aureus in mice can establish infection even in the presence of vancomycin, and we have developed a novel therapeutic that effectively kills intracellular S. aureus. This antibody‐antibiotic conjugate consists of an anti‐S. aureus antibody that recognizes a highly expressed epitope on wall teichoic acid, conjugated to a potent antibiotic that is activated only after its release from the antibody in the proteolytic environment of the phagolysosome. The antibody‐antibiotic conjugate is superior to vancomycin for treatment of bacteremia and provides direct evidence that intracellular S. aureus represents an important component of invasive infections. This paradigm may have implications for treatment of other bacterial infections.Support or Funding InformationThis work was funded by Genentech, Inc.
Engineered bacteriophage therapeutics against multidrug‐resistant pathogens
Authors: Barbu Magda , DelRosario Yvette , Hubby Bolyn 
Published: November 2023

Since the introduction of penicillin over 70 years ago, the number of patient deaths caused by infectious diseases has dramatically decreased and antibiotics have become among the most commonly prescribed drugs. However, the selective pressure imposed by antibiotic overuse has led to the emergence of bacterial strains resistant to virtually all antimicrobials and has renewed interest in the development of lytic bacteriophage (or phage) therapeutic and prophylactic agents. The advantages of using phage to combat infectious diseases include minimal disruption of resident flora, lack of cross‐resistance with antibiotics, low toxicity and self‐limiting dosing. Despite these attractive features, translational development of natural phage has been hindered mainly by the difficulty of accessing bacterial hosts within biofilms, the rapid emergence of resistant bacteria to a single phage, and above all, by the narrow host specificity of phage compared to antibiotics. The need for customized and complex combinations of natural phage to achieve adequate host range activity has made their development as licensed therapeutics very difficult. Genetic engineering of phage genomes can overcome these hurdles; however, broadly applicable methods for efficient construction of defined mutations in virulent phage genomes are still in their infancy. Thus, we developed a completely cell‐free phage engineering method that allows rapid and iterative editing of viral genomic DNA. In parallel, we sequenced and annotated the genomes of ~300 MDR P. aeruginosa clinical isolates, and subsequently determined the susceptibility of each isolate to specific phages. Through bioinformatics analysis in combination with our engineering platform, we have successfully collapsed the host range of a family into a representative phage. We further engineered wide host range phage to express secondary payloads, such as biofilm degrading enzymes and antimicrobial moieties, and demonstrated that these phages have improved activity compared to natural phage in P. aeruginosa models of infection.Support or Funding InformationSynthetic Genomics Vaccines, Inc.NIAID R21AI121531
L‐Sulforaphane Decreased Contractile Response in Mesenteric Arteries in a Rat Model of Gestational Hypertension
Authors: Cushen Spencer C , Osikoya Oluwatobiloba , Goulopoulou Styliani 
Published: November 2023

BackgroundMaternal hypertension is a state of inflammation characterized by oxidative stress. Exposure of pregnant rats to the Toll‐like Receptor 9 activator, ODN2395, induces hypertension and upregulates vascular oxidative stress. The transcription factor, Nuclear Factor, Erythroid 2 Like 2 (Nrf2), is a regulator of antioxidant response, which is overexpressed in placentas from patients with preeclampsia. However, the role of Nrf2 in maternal vascular dysfunction is unknown.HypothesisL‐sulforaphane, an Nrf2 activator, will have anti‐contractile effects on arteries from pregnant rats treated with CpG oligonucleotides (a model of gestational hypertension).MethodsPregnant Sprague‐Dawley rats were treated with synthetic unmethylated CpG oligonucleotides (ODN2395, 100μg/intraperitoneal injection) or saline (Control) on gestational day 14, 16, and 18 (term=21–22 days). Blood pressure was measured before pregnancy and on gestational day 19 using the tail cuff method. The contractile responses of mesenteric resistance arteries to a thromboxane A2 (TxA2) mimetic, U46619, in the presence or absence of Nrf2 activator, L‐sulforaphane (L‐S, 40 μM), were assessed by wire myography on gestational day 21.ResultsDams treated with ODN2395 had greater systolic blood pressure on gestational day 19 compared to control rats (Control, n=9: 100±4 mmHg vs. ODN2395, n=7: 119±4 mmHg, p=0.007). One‐hour incubation with L‐sulforaphane did not affect contractile responses to U46619 in arteries from control or ODN2395‐treated rats. Incubation with L‐sulforaphane for 3 hours reduced the contractile response to U46619 in mesenteric arteries from both ODN2395 and control rats (Peak contraction as %Max KCl (120mM), Control Veh: 120.5%±4.85; Control L‐S: 39.1%±7.16; ODN2395 Veh: 111.1%±4.19; ODN2395 L‐S: 42.6%±2.68).ConclusionPregnancy is a state of oxidative stress and this may explain the anti‐contractile effects of L‐sulforaphane in arteries from normal, healthy rats. In preeclampsia, levels of oxidative stress are greater compared to normotensive pregnancies and thus, systemic treatment with L‐sulforaphane or other Nrf2 activators may improve poor cardiovascular outcomes in pregnancies with preeclampsia.Support or Funding InformationUniversity of North Texas Health Science Center; American Heart Association
Arterial Pulse Wave Velocity is Increased in Prematurely‐Born Adults and Rats Given Neonatal Supplemental Oxygen
Authors: Peters Matthew , Hoover Michael , Sturgeon Madison , Vellookunnel Shilpa , Chandrasekar Shreya , Schaeffer Gilbert V. , Sindler Amy L. , Pierce Gary L. , Stauss Harald M. , Farrell Emily T. , Eldridge Marlowe W. , Bates Melissa 
Published: November 2023

Thirteen percent of infants are born prematurely and this population is at increased risk of cardiovascular disease (CVD) as they age. We hypothesized that compared with term‐born controls (CON), individuals born prematurely would have premature aortic stiffening, indicating greater CVD risk.To test this hypothesis in humans, aortic pulse wave velocity (aPWV) was used to determine aortic stiffness in young adults born preterm between 1989 and 1991 (PT, 21–23 years) and age‐matched controls (CON). Intra‐arterial brachial artery pressure waveforms from 9 PT and 11 CON subjects were converted to central aortic pressure waveforms using a published transfer function. aPWV was then derived from the time delay between the forward and reflected waves and the effective reflecting distance, estimated based on individual anthropometric data. To test the hypothesis in a rodent model that perinatal supplemental oxygen (O2) promotes aortic stiffening, newborn rat pups were exposed to 80% O2 for 14 days, or 21% O2 as a control.aPWV was higher in PT (10.1±2.6 m/s) compared with CON (7.2±0.8 m/s, p=0.025). aPWV did not correlate with systolic blood pressure in either group (PT R2=0.02, p=0.73; CON R2=0.17, p=0.20). At age 12 months, aortic stiffness, determined by aPWV, was higher in 80% O2‐exposed rats (487.1±27.8 cm/s) compared with controls (371.1±13.25 cm/s, p=0.02).These data suggest that individuals born prematurely experience accelerated vascular aging of the aorta and that exposure to supplemental O2 as part of clinical care in premature infants may be a potential mechanism mediating this novel observation.Support or Funding InformationUniversity of Iowa Start‐Up Fund
Smooth Muscle Cells Are the Source of Heart Failure‐Causing Methylglyoxal in Diabetes Mellitus
Authors: Bidasee Keshore R , Alomar Fadhel , Fang Rui , Cox Jesse L , Rozanski George J , Singh Jaipaul 
Published: November 2023

Heart failure/diabetic cardiomyopathy (DC) is an established case of morbidity and mortality in individuals with diabetes mellitus (DM). Yet, its molecular triggers remain undefined. Supra‐physiologic levels of the dicarbonyl species methylglyoxal (MG) has emerged as a leading candidate since it triggers several of the pathobiologies reported in DC. Why/how MG increases in the heart in DM remains undefined. Here left ventricular tissues from DM patients (Type 1 and Type 2 DM) and Type 1 DM rats were used to assess protein levels of the MG synthesizing and degradation enzymes, vascular adhesion protein‐1 (VAP‐1) and glyoxalase‐I (Glo‐I), respectively. Type 1 DM rats were also used to determine if targeted increase of Glo‐I in the heart would be cardio‐protective. In left ventricular tissues from patients and rats with DM, VAP‐1 was upregulated in smooth muscle cells (SMCs) and Glo‐I was reduced in SMCs and ventricular myocytes. MG and its metabolite argpyrimidine were also upregulated in left ventricular tissues. Increasing Glo‐I protein in SMCs and myocytes of Type 1 DM rats using a custom‐designed adeno‐associated virus blunted myocyte dysregulation and DC progression. Glo‐I restoration also attenuated VAP‐1 upregulation, lowered MG production, prevented microvascular leakage, and reduced mitochondria ROS generation, NF‐κB activation and TNF‐α production. It also blunted argpyrimidine modification and dysregulation of the sarcoplasmic reticulum Ca2+ cycling proteins, RyR2 and SERCA2a. From these new data we conclude that the myocyte‐dysregulating and heart failure‐causing increase in MG during DM arises from increased expression of VAP‐1 in ventricular SMCs and from decreased expression of Glo‐I in SMCs and ventricular myocytes. They also suggest that lowering SMCs‐generated MG could also be a new therapeutic strategy to blunt heart failure/DC development in DM.Support or Funding InformationGTRP 1053, UNMC, UNeMED
Cavin‐3 deficiency reduces caveolae abundance in vascular and urinary bladder smooth muscle cells
Authors: Zhu Baoyi , Swärd Karl , Ekman Mari , Uvelius Bengt , Rippe Catarina 
Published: November 2023

We investigated the effect of cavin‐3 (protein kinase C delta binding protein, Prkcdbp) deficiency on vascular and urinary bladder function. Small mesenteric arteries and the urinary bladder were isolated from mice lacking cavin‐3 (Prkcdbp−/−, knockout: KO) and from control littermates (Prkcdbp+/+, wild type: WT). One group of mice was used for blood pressure measurements, whereas another set of mice underwent bladder outlet obstruction followed by analysis of protein expression using western blotting. Our studies revealed that loss of cavin‐3 resulted in a 40% reduction of the caveola protein cavin‐1 (polymerase I and transcript release factor, Ptrf) in vascular and bladder smooth muscle. Electron microscopy demonstrated that loss of cavin‐3 was accompanied by a reduction of caveolae by 40% in smooth muscle, while the abundance of caveolae in endothelial cells was unchanged. Vascular contraction in response to cirazoline (α1‐adrenergic agonist) was normal, but nitric‐oxide dependent relaxation was enhanced. The latter effect was associated with elevated expression of soluble guanylyl cyclase, and yet blood pressure was similar in WT and KO mice. Contraction of the urinary bladder was not affected by loss of cavin‐3, and the proteomic response to outlet obstruction, including STAT3 (signal transducer and activator of transcription 3) phosphorylation, induction of synthetic markers, and repression of contractile markers were identical, the only exception being a curtailed induction of the Golgi protein GM130 (Golgi matrix protein 130 kD). In conclusion, loss of cavin‐3 thus reduces the number of caveolae in smooth muscle and partly destabilizes cavin‐1, but the functional consequences are modest and include an elevated vascular sensitivity to nitric oxide and a somewhat disturbed Golgi homeostasis in a situation of severe cellular stress.Support or Funding InformationThis study was funded by The Swedish Research Council (Vetenskapsrådet, http://www.vr.se/, K2015‐65X‐22662‐01‐3/KS), the Crafoord Foundation (http://en.crafoord.se/, CR), and the Royal Physiographic Society (Kungliga Fysiografiska Sällskapet, http://www.fysiografen.se/, CR).
Transient Receptor Potential Vanilloid 4 Channel Mediates Endothelium‐Dependent Vasodilation in Parenchymal Arterioles
Authors: Diaz‐Otero Janice M , Jackson William F , Dorrance Anne M 
Published: November 2023

Chronic hypertension increases the risk of vascular dementia development and damage caused by cerebral ischemia. This may be partially the result of impaired endothelium‐dependent dilation of parenchymal arterioles (PAs) that regulate perfusion of the white matter. Transient receptor potential (TRP) channels such as the TRP vanilloid 4 (TRPV4) regulate endothelium‐dependent vasodilation and calcium homeostasis in peripheral arteries and large cerebral arteries. However, the role of TRPV4 in PA vasodilation has not been studied. We hypothesized that TRPV4 activation is a key regulator of endothelium‐dependent dilation in the PAs from normotensive and hypertensive rats. PAs vasodilation from 14–15 week old male normotensive Sprague Dawley (SD) rats and stroke‐prone spontaneously hypertensive rats (SHRSP) were studied by pressure myography. Data are presented as mean ± SEM. The presence of TRPV4 in the PAs was confirmed by immunostaining. The PAs from SHRSP had increased myogenic tone (30±5 vs 51±6 %tone, n=8; SD vs SHRSP; p = 0.02) and impaired carbachol (CbCh)‐induced dilation (35±7 vs 25±7 %dilation, n=4; SD vs SHRSP; p = 0.001) compared to the SD rats. After the development of myogenic tone (60 mmHg) PAs were incubated with the TRPV4 antagonist, GSK2193874 (GSK2, 10−7M) for 10 minutes, then CbCh‐mediated dilation (10−9–10−5M) was assessed. In normotensive rats, TRPV4 antagonism prevented the CbCh‐induced vasodilation (54±7 vs −7±14 %dilation at 10−5M, n = 4; CbCh vs CbCh+GSK2; p =0.008). In the SHRSP, inhibition of TRPV4 caused a loss of myogenic tone in the PAs (46±9 vs 23±12 %tone, n = 4; before vs. after GSK2; p = 0.1) that was not observed in SD rats, but this was not significantly different. TRPV4 antagonism also prevented endothelium‐dependent vasodilation in the PAs from SHRSP (46±1 vs −30±20 %dilation at 10−5M, n = 4; CbCh vs CbCh+GSK2; p = 0.02). These data show that TRPV4 plays an important role in mediating endothelium‐dependent dilation in cerebral arterioles from normotensive and hypertensive rats. TRPV4 may also contribute to augmented myogenic tone in SHRSP PAs, a role not observed in PAs from SD rats; this could be due to changes in the expression or activity of TRPV4 in the smooth muscle cells. Our data suggest that TRPV4 may have a critical role in mediating the hypertension‐associated changes in the vascular function of cerebral arterioles. The impaired vasodilation and changes in myogenic tone may compromise the regulation of cerebral blood flow during hypertension. TRPV4 could be a potential therapeutic target to improve vascular function of cerebral arterioles during hypertension and to decrease the risk of stroke and vascular dementia development that accompany hypertension.Support or Funding InformationSupported by PO1‐HL070687
Effects of High Glucose and Major Metabolic Substrates on Alpha‐adrenergic Contractions in Rat Mesenteric and Deep Femoral Arteries
Authors: Yoo Hae Young , Vorn Rany 
Published: November 2023

Hyperglycemia causes vascular damage and increased risk of cardiovascular diseases. It has been demonstrated that chronic exposure to high glucose impaired endothelial functions. However, specific effects of short‐term exposure to high glucose on vascular reactivity are controversial. Moreover, the combined effects of other metabolic substrates such as free fatty acids (FFA) on vascular reactivity remain poorly understood. Here we investigate the effects of short‐term exposure to high glucose with or without major metabolic substrates including FFAs termed “nutrition with fatty acid” (NF) solution, on mesenteric (MA) and deep femoral arteries (DFA) of rats. Arterial ring segments were mounted in a double‐wire myograph. Contraction in response to phenylephrine (PhE) was determined in control (5 mM) and high glucose (23 mM, HG) environments over a 30 min period. Our results showed that PhE‐induced vasocontraction was enhanced by pre‐incubation of HG solution in both arteries. A combined incubation with HG and palmitic acid (100 mM) induced similar sensitization of PhE‐contractions in both arteries. In contrast, high K+‐induced contractions were not affected by HG. Interestingly, pre‐incubation with NF solution decreased PhE‐induced contraction in DFA but increased the contraction in MA. In NF solution, the HG‐induced facilitation of PhE‐contraction was not observed in MA. Furthermore, the PhE‐induced contraction of DFA was attenuated by HG in NF solution. Our results demonstrate that the sensitization of PhE‐induced arterial contraction by HG is differentially affected by other metabolic substrates. The conversation of skeletal arterial contractility by HG in NF solution requires careful interpretation of the previous in vitro studies where only glucose is included in physiological salt solutions. Further studies are required to elucidate the mechanism underlying the inconsistent effect of NF solution on MA and DFA.Support or Funding InformationThis study was supported by a National Research Foundation of Korea grant funded by the Ministry of Science, ICT, & Future Planning (2015R1C1A1A01054038).
Impaired autophagy contributes to the vascular dysfunction in type 2 diabetes
Authors: Choi Soo‐Kyoung , Byeon Seonhee , Kwon Youngin , Lee Young‐Ho 
Published: November 2023

Vascular dysfunction is a major complication in type 2 diabetes. It has been suggested that dysregulation of autophagy is associated with various cardiovascular diseases. However, the relationship between autophagy and type 2 diabetic vascular dysfunction remains unclear. Thus, in this study, we examined whether impaired autophagy is involved in vascular dysfunction of type diabetic mice. Ten to 12‐week old male type 2 diabetic (db−/db−) mice and their control (db−/db+) mice were used. The expressions of LC3□ and beclin‐1 (markers of autophagy) were decreased in mesenteric artery, aorta and heart from type 2 diabetic mice compared to control mice.Support or Funding InformationThis work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Science, ICT & Future Planning) (No. 2016R1C1B1006504).
Comparisons of Proximal Aortic Pulse Wave Velocity Measurements: Carotid‐Femoral vs. Heart‐Brachial PWV
Authors: Sugawara Jun , Tomoto Tsubasa , Tanaka Hirofumi 
Published: November 2023

The proximal aorta (e.g., the ascending aorta and aortic arch) is the most elastic segment of the arterial tree directly connected with the left ventricle and plays the dominant role in buffering the cyclic mechanical forces of cardiac pulsations. Carotid‐femoral pulse wave velocity (cfPWV) is considered as the reference standard measurement for large elastic artery stiffness but the most proximal segments of the aorta are omitted in its measurement. The primary aim of the present study was to compare two indices of arterial stiffness that differ in the contribution of proximal aortic segments. To do so, using the cross‐sectional study design, we compared cfPWV (mainly reflecting stiffness of the abdominal aorta) and heart‐brachial PWV (hbPWV; including the proximal aortic stiffness) in relation to aging‐related change and to central hemodynamic indices. In a total of 190 adults (92 men and 98 women) aged 53±15 yrs were studied. Pulse transition times were obtained between carotid and femoral pressure waveforms (via applanation tonometry) for cfPWV and between the second heart sound and the dicrotic notch of brachial pressure waveforms (via air‐plethysmography) for hbPWV, respectively. Arterial path lengths were measured directly by MRI. Central aortic hemodynamic indices were estimated from carotid arterial pressure waveform by general transfer function‐based pulse wave analysis. cfPWV and hbPWV were significantly correlated with age (r=0.68 and r=0.79, both P<0.0001). Aging‐related increases (calculated from the slope of regression lines) were 9.9%/decade in cfPWV and 14.7%/decade in hbPWV. Aortic augmentation index (an index of left ventricular after load) was significantly correlated with cfPWV (r=0.45) and more strongly with hbPWV (r=0.50). Likewise, aortic systolic tension‐time integral and diastolic pressure‐time integral (indices of myocardial oxygen demand and supply) were correlated with hcPWV (r=0.49 and r=0.53, P<0.0001 for both) and more strongly with hbPWV (r=0.50 and r=0.57, P<0.0001 for both). Our findings suggest that a measure of PWV including the ascending aorta exhibits a greater age‐related increase and stronger associations with central hemodynamic indices than cfPWV. Prospective data linking hbPWV to clinical endpoints are needed.Support or Funding InformationThis study was supported by JSPS KAKENHI Grant Number JP25702045 and JP26670116.