bluuebunny/crossref_metadata_2025_split huggingface dataset:
DPO‐1‐sensitive K+ channels control the tone and reactivity of resistance‐sized arterioles from brain and skeletal muscle (678.5)
Authors: Butcher Joshua , Fancher Ibra , Frisbee Jefferson , Dick Gregory 
Published: June 2021

Arterial smooth muscle cells possess prominent delayed rectifier K+ (KDR) currents. KDR currents may be mediated by KV1.5 channels, which are sensitive to inhibition by DPO‐1. We tested the hypothesis that DPO‐1‐sensitive KDR channels control the tone and reactivity of resistance‐sized arterioles from brain and skeletal muscle. Rat (n = 4) middle cerebral and gracilis arterioles were dissected, cannulated, and pressurized to 80% of mean arterial pressure (86 ± 3 mmHg) for video microscopy measurements. With active tone, the diameters of gracilis and cerebral arterioles were 107 ± 5 and 103 ± 4 μm. DPO‐1 (10 μM) constricted gracilis and cerebral arterioles to 79 ± 4 and 73 ± 5 μm. DPO‐1 increased the sensitivity (logEC50) to phenylephrine (6.3 ± 0.1 vs 6.9 ± 0.1) and serotonin (6.5 ± 0.1 vs 7.2 ± 0.1) in gracilis and cerebral arterioles, respectively. Increasing internal pressure from 40 to 140 mmHg in 20 mmHg steps activated myogenic responses (diameter decreased 0.18 ± 0.05 and 0.17 ± 0.05 μm/mmHg in gracilis and cerebral arterioles). DPO‐1 eliminated myogenic responsiveness, as diameter increased 0.08 ± 0.07 and 0.06 ± 0.05 μm/mmHg. Dilation to 100 μM acetylcholine was 27 ± 3 and 27 ± 1 μm in gracilis and cerebral arterioles; DPO‐1 reduced these dilations to 4 ± 2 and 10 ± 2 μm. We suggest that DPO‐1‐sensitive KDR channels (KV1.5) play a major role in determining microvascular tone and responses to vasoconstrictors and vasodilators.Grant Funding Source: Supported by T32HL090610
Relief of endoplasmic reticulum stress in the brain rescues high fat diet‐induced hypertension but this is not due to the subfornical organ (682.3)
Authors: Young Colin , Butler Scott , Horwath Julie , Mark Allyn , Davisson Robin 
Published: June 2021

We have recently demonstrated robust ER stress in central neural cardioregulatory regions of HFD animals, with particularly prominent elevations in the SFO. In line with this, global relief of brain ER stress via intracerebroventricular delivery of the chemical ER chaperone TUDCA ameliorates HFD‐induced hypertension (Δ‐6±1 mmHg, p<0.05). However, the precise neural regions involved remain unknown. We hypothesized that ER stress in the SFO mediates obesity‐induced hypertension. HFD (n=13; 20 wks) and age‐matched normal chow (n=10) fed C57Bl/6 mice were instrumented with radiotelemeters for mean arterial pressure (MAP) measurements and underwent SFO‐targeted microinjections of an adenovirus encoding the ER chaperone GRP78 (AdGRP78) to reduce ER stress. Relative to normal chow, HFD control vector treated mice demonstrated a significant increase in MAP (102±3 vs 116±1 mmHg, normal chow vs HFD, p<0.05). SFO‐targeted AdGRP78 did not influence the HFD‐induced increase in MAP (118±1 mmHg, HFD AdGRP78, p>0.05). Analysis of ER stress biomarkers revealed that AdGRP78 selectively reduced ER stress within the SFO, but not other neural regions (e.g. p58IPK mRNA SFO: 1.0±0.1 vs 0.6±0.1, HFD AdLacZ vs HFD AdGRP78, p<0.05). These findings suggest that, while brain ER stress contributes to HFD‐induced hypertension, ER stress in the SFO does not appear to be involved in the maintenance of obesity‐mediated hypertension.Grant Funding Source: Supported by HL63887, HL96571, HL84207, AHA13POST14410020, K99HL166776
Dynamic regulation of microRNA networks in the brainstem underlie hypertension development (686.22)
Authors: DeCicco Danielle , Zhu Haisun , Brureau Anthony , Schwaber James , Vadigepalli Rajanikanth 
Published: June 2021

This study aims to elucidate the mechanisms by which microRNAs (miRNA) regulate autonomic control networks driving hypertension development. Using a high‐throughput assay for 419 miRNAs in the Spontaneously Hypertensive Rat (SHR) and Wistar‐Kyoto (WKY) control at three key stages of hypertension development, significant differences appear in 23 miRNAs of the nucleus tractus solitarius (NTS) and rostral ventrolateral medulla (RVLM). Alterations were stage‐ and region‐dependent, with significant differences occurring at mid‐stage in NTS vs early‐stage in RVLM. 96 gene targets corresponding to catecholaminergic processes and neuromodulation were predicted using the miRWALK database and tested using high‐throughput qPCR. Using correlational relationships between miRNAs and mRNAs, we observed a double‐negative regulatory motif, which appears prevalent throughout the data. This motif consists of a miRNA down‐regulating a negative regulator of a pro‐hypertensive signaling pathway like Angiontensin II signaling or leukotriene‐based inflammation. We demonstrated for the first time that the broad concordance of microRNA dynamics and target gene expression compose a regulatory network in the brainstem underlying hypertension.Grant Funding Source: Supported by NIGMS R01 GM083108, NHLBI R01 HL111621
Central projections of sodium‐sensitive afferent renal nerves in rats (687.3)
Authors: Goodwill Vanessa , Loewy Arthur , Knuepfer Mark 
Published: June 2021

Renal denervation is reported to treat resistant essential hypertension. Many suggest that afferent renal nerves (ARN) are responsible for the full expression of hypertension. We hypothesized that osmosensitive ARN activate sites in the CNS that contribute to hypertension by evoking excessive sympathetic nerve activity. To determine the central projections, a dual lumen cannula was implanted through the left ureter into the renal pelvis of the rat. Several days later, renal pelvic afferents were bathed with different concentrations of NaCl (20, 308 and 500 mEq/L) using a push‐pull perfusion for 2 hrs while recording arterial pressure in conscious rats. Activation of CNS sites was identified using immunohistochemical detection of c‐Fos protein and compared to controls (no infusion or denervation). Many sites were dose‐dependently stimulated in response to intrarenal infusion of NaCl. These included the nucleus tractus solitarius, area postrema, parabrachial nucleus, paraventricular hypothalamus, and other areas. The results indicate that stimulation of osmosensitive ARN leads to activation of central autonomic sites involved in the regulation of sympathetic outflow. Thus, this work identifies likely sites mediating ARN‐induced sympathoexcitation contributing to the pathogenesis of hypertension.Grant Funding Source: Supported by USPHS HL25449 (ADL) and DA017371 (MMK).
SMAD1 signaling mediates inhibition of store‐operated Ca2+ entry on collagen IV expression in glomerular mesangial cells (689.14)
Authors: Wu Peiwen , Wang Yanxia , Chaudhari Sarika , Zuckerman Jonathan , Davis Mark , Ma Rong 
Published: June 2021

Objective: The Smad signaling pathway plays a critical role in extracellular matrix protein production in glomeruli in diabetic nephropathy. Store‐operated Ca2+ entry (SOCE) regulates protein synthesis. The aim of the present study was to determine if SOCE in mesangial cells (MCs) suppressed production of collagen type IV (Col IV) by inhibiting Smad1.Methods: Western blot and immunohistochemistry were used to detect target protein expression in cultured MCs and mouse glomeruli, respectively. Systemic administration of nanoparticles containing siRNA against Orai1 was employed to deliver the siRNA to MCs in mice.Results: In cultured human MCs, activation of SOCE by cyclopiazonic acid or thapsigargin (TG) significantly suppressed Ang II‐induced Smad1 phosphorylation. Overexpression of STIM1 dramatically decreased abundance of Smad1 protein. Furthermore, high glucose‐induced Col IV protein expression was significantly inhibited by TG. Inhibition of SOCE by 2‐aminoethoxydiphenyl borate or La3+ increased Col IV expression which was accompanied by an increase in Smad1 protein expression. However, the Col IV response to SOCE inhibition was not observed in MCs with knockdown of Smad1. Moreover, in vivo knockdown of Orai1 in mouse MCs increased expressions of both Smad1 and Col IV in the nanoparticle‐positive glomeruli.Conclusion: SOCE inhibits Col IV expression by inhibiting Smad1 pathway in MCs.Grant Funding Source: NIH/NIDDK
Multilevel synchronization of tubuloglomerular feedback is modulated by nitric oxide (692.10)
Authors: Mitrou Nick , Scully Christopher , Braam Branko , Chon Ki , Cupples William 
Published: June 2021

Previously we showed that NO modulates synchronization of both mechanisms of renal autoregulation. We did not, however, provide a mechanism to explain TGF synchronization after L‐NAME. L‐NAME changed TGF synchronization in a way we did not understand. Here we show that TGF synchronization may become stronger within cortical lobules after L‐NAME, resulting in two mechanisms of synchronization within a synchronized cluster. Male Long‐Evans rats (N=8) were anaesthetized (isoflurane) and their left kidneys were exposed. Cortical perfusion during control and NOS inhibition (L‐NAME, 10mg/kg i.v.) was monitored by laser speckle perfusion imaging. The TGF frequency range (0.01‐0.06 Hz) was isolated with a band pass filter. Clusters of synchronized nephrons were identified based on phase coherence (PC) between all pairs of imaged pixels. During control, TGF was synchronized moderately over large areas (~5x7 mm). After L‐NAME, each cluster had areas with high PC (strong synchronization) in the center and lower, but significant, PC in the surround. When the PC reference pixel was moved systematically there was strong synchronization within smaller ~800 x 800 um areas. The position of the high PC areas was consistent with an anatomical basis for synchronization. We conclude that TGF synchronization becomes stronger within cortical lobules while remaining weakly synchronized among lobules within the larger clusters.Grant Funding Source: Supported by CIHR MOP‐102694
Biotin ligase tagging identifies E‐cadherin protein neighbors, including lipoma preferred partner, which modulates epithelial cell‐cell and cell‐substrate adhesion (694.4)
Authors: Van Itallie Christina , Tietgens Amber , Aponte Angel , Fredriksson Karin , Fanning Alan , Gucek Marjan , Anderson James 
Published: June 2021

Proteins known to be associated with cellular adherens junctions (AJs) include E‐cadherin, catenins and other proteins involved in actin organization, vesicular trafficking and various signaling pathways. However, it is likely that some proteins with important roles at the AJ have not yet been identified. To detect additional AJ proteins, we expressed E‐cadherin fused to an engineered biotin ligase in MDCK epithelial cells and purified proximal, biotinylated proteins; these were identified by mass spectrometry. Catenins were among the most abundant of the 300+ proteins identified, along with nearly 40 other proteins previously reported to influence the AJ functions. Other proteins could be easily understood as novel candidates for regulating the structure or function of the AJ. One protein, lipoma preferred partner (LPP) was chosen for further characterization. Knockdown of LPP in MDCK cells demonstrated it was required for normal E‐cadherin‐dependent adhesion and was likely involved in coordination of cell‐cell and cell‐substrate adhesion. The demonstration of a role for LPP in E‐cadherin‐dependent adhesion was a proof of principle that the analysis of E‐cadherin protein neighbors could generate new tools to aid in AJ investigation.Grant Funding Source: Supported by the Intramural Research Program of the NIH.
N‐cadherin adhesion mediated signaling stabilizes VE‐cadherin‐mediated adhesion and strengthens endothelial barrier function (695.3)
Authors: Kruse Kevin , Sieracki Nathan , Komarova Yulia 
Published: June 2021

N‐cadherin adhesion mediated signaling stabilizes VE‐cadherin‐mediated adhesion and strengthens endothelial barrier function Kevin Kruse, Nathan Sieracki, Yulia Komarova.Neuronal (N)‐cadherin has been shown to be involved in maintaining endothelial barrier function by forming homophilic interaction between endothelial cells (ECs) and pericytes, although it is unknown whether this is solely due to recruitment of pericytes to the endothelium or whether there are signaling events downstream of N‐cadherin homophilic adhesions in ECs that may play a role in strengthening the endothelial barrier. Our research objective therefore is to understand the role of N‐cadherin adhesion mediated signaling in regulating endothelial barrier function. Because N‐cadherin does not form homotypic interactions in an in vitro cell culture monolayer, we have utilized surface chemistry techniques to mimic the N‐cadherin interactions found between ECs and pericytes in vivo. The extracellular domain of N‐cadherin was covalently attached to an Ni‐NTA cover slip, allowing us to study N‐cadherin‐mediated adhesion in ECs. Our data show that ECs grown on N‐cadherin coated surfaces have increased VE‐cadherin accumulation as well as decreased t½ (half‐time to full fluorescence recovery) of VE‐cadherin turnover at cell‐cell junctions as determined by immunofluorescent staining and fluorescent recovery after photobleaching (FRAP). Our data indicate that N‐cadherin outside‐in signaling promotes formation of VE‐cadherin‐mediated adhesion by possibly stabilizing VE‐cadherin trans‐interactions. We will further use VE‐ or N‐cadherin biomimetic surfaces to isolate either VE‐cadherin or N‐cadherin complexes in order to understand the common and distinct binding partners and signaling pathways downstream of VE‐ and N‐cadherin.Grant Funding Source: Supported by NIH T32
Experience for using platelet‐rich plasma in the treatment of androgenic alopecia (695.8)
Authors: Batpenova Gulnar , Sadykova Gulnaz , Kotlyarova Tatiyana 
Published: June 2021

One of new physiological and correspondingly safe treatment in dermatology is PRP (PRP). Literature data suggest a positive PRP role in hair growth.Objective: to evaluate the efficacy and safety of monotherapy PRP in the treatment of androgenetic alopecia.Material and Methods: 15 patients with androgenetic alopecia , including male ‐9, 6 ‐ women aged 21 to 58 years , average age ‐ 40.2 years. Duration of disease, on the average is 6.5 years. Monotherapy was carried out by intradermal injection of platelet rich plasma at intervals of 2 months. The number of procedures is 1‐4, on the average ‐ 2. Evaluation of the results was made by macrofotografic way. The observation period is 12 months.Results: as a result of platelet‐rich plasma therapy, there is a positive trend established as the form of hair loss reduction and enhancing their growth. Patients receiving more than 3 treatments achieved the significant effect. During and after treatment the side effects for a year have not been noted.Conclusions: The results of the treatment are definitely confirmed by the macrophotografic data. However, the present study requires a careful study the influence of platelet rich plasma, not only to the growth of hair, but also on the structure, which suggests to conduct more extensive controlled studies using methodics such as dermoscoy, video dermoscopy, trihoscopy, determination Ki‐67 markers in a large cohort of patients.
Elevated carboxyhemoglobin in a marine mammal (698.1)
Authors: Tift Michael , Ponganis Paul , Crocker Daniel 
Published: June 2021

Low concentrations of endogenous carbon monoxide (CO), generated through catabolism of heme‐proteins, have been shown to protect tissues against inflammatory diseases, and ischemia‐reperfusion injuries. However, elevated carboxyhemoglobin (COHb) formed by CO binding to hemoglobin, prevents adequate O2 delivery to tissues by lowering arterial O2 content. Therefore there must be a balance in the level of endogenous CO which maximizes cytoprotective effects and minimizes inhibition of O2 delivery. The elevated heme‐protein concentrations as found in marine mammals, are likely associated with increased heme catabolism, and subsequently increased endogenous CO production and COHb values. We measured COHb in elephant seals, a species with exceptional blood volumes, hemoglobin concentrations and myoglobin content. The mean value in adult seals was 8.7 ± 0.3%, while juveniles and pups (with lower heme‐protein stores) had COHb values of 7.6 ± 0.2% and 7.1 ± 0.3%, respectively. Serial samples over several hours revealed little to no fluctuation in COHb values. This consistent elevation in COHb suggests the magnitude and/or rate of heme‐protein turnover, is higher than terrestrial mammals. While these COHb values decrease the O2 stores needed for diving, the constant presence of elevated CO in blood may provide protection against ischemia‐reperfusion events associated with breath‐holds in elephant seals.Grant Funding Source: Supported by National Institute of Health NHLBI RO1‐HL091767
What is the role of S‐nitrosation of soluble guanylyl cyclase in the heart? (698.9)
Authors: Huang Can , Crassous Pierre‐Antoine , Beuve Annie 
Published: June 2021

Nitric oxide (NO) is involved in cardioprotection against ischemia/reperfusion (I/R) injury. Soluble guanylyl cyclase (sGC) produces cGMP when it is stimulated by NO. Association of the α and β subunit of sGC is required for cGMP‐forming activity. NO protects the heart from I/R via a cGMP‐dependent and a S‐nitrosation (SNO)‐dependent pathway. Preliminary evidence showed that sGC‐overexpression increases S‐nitrosation of specific proteins (SNO‐proteins) in rat neonatal cardiomyocytes. These sGC‐dependent SNO‐proteins were identified by Mass Spectrometry and are mostly associated with intercalated disc and cytoskeleton, including integrin‐linked kinase (ILK) and desmin. sGC is heavily S‐nitrosated in the presence of nitrosating agent. Thus, we hypothesize that sGC acts as a transnitrosylase in the heart, e.g. transfers its SNO to specific proteins. Intriguingly, sGC‐α and β subunits localize differently in cardiomyocytes. β is at the nuclear envelope and sarcolemma, while α is in cytosol, suggesting that each subunit might have a separate function, independent of cGMP‐forming activity. In cardiomyocytes treated with a nitrosating agent, overexpressing a single sGC subunit, either α or β, was sufficient to increase SNO level of ILK and desmin. Hence, we propose that sGC could modulate cardioprotection by increasing protein‐SNO, in addition to the NO‐cGMP dependent pathway.Grant Funding Source: Supported by 6R01 GM067640
PPAR‐gamma is required in the myocardium for outflow tract development (699.2)
Authors: Keith Benjamin , Zhou Lun , Xie Linglin 
Published: June 2021

One‐third of congenital heart defects involve abnormalities in the development of the outflow tract (OFT). In our lab’s previous study, 100% of PPAR‐gamma heterozygote mouse embryos at embryonic day 14.5 (E14.5) showed OFT defects including ventricular septal defects (VSD), overriding aorta (OA), double outlet right ventricle (DORV), and common arterial trunk (CAT). These observations suggested that PPAR‐gamma plays a vital role in the development of the OFT. To identify the destined cell lineage for PPAR‐gamma in a specific cell lineage, we performed cre‐lox recombination technology based genetic study to knockdown PPAR‐gamma in specific cell lineages including the myocardium (Tnt‐cre), endocardium (Tie2‐cre), and the second heart field (Mef2c‐cre). We found that 33% (2/6) PPAR‐gammaTnt‐cre/+ embryos at E13.5 had OA, but no OFT defects were observed in either PPAR‐gammaTie2‐cre/+ or PPAR‐gammaMef2c‐cre/+. This data revealed that PPAR‐gamma is required in the myocardium, but not endocardium nor second heart field, for the development of the OFT.Grant Funding Source: Supported by the APS STRIDE summer undergraduate fellowship and the UND faculty seed money
Time in culture alters the biophysical responses of rat carotid body cells (709.1)
Authors: Kim Insook , Donnelly Dave , Fite Lasha , Carroll John 
Published: June 2021

The carotid body (CB) senses blood O2, CO2, and pH. Fresh CB glomus cells depolarize and increase intracellular Ca2+ ([Ca2+]i) in response to K+, and hypoxia but not to ATP, while non‐glomus cells increase [Ca2+]i in response to extracellular ATP, alone. We determined whether these response characteristics changed in culture. Cells dissociated from postnatal day 15 rat CB were plated and cultured for 7‐8 days. Cultured CB cells were tested for [Ca2+]i responses to 20mM K+, hypoxia, anoxia, TEA(10mM)+4‐AP(5mM), and ATP(100µM) when fresh (0 day culture, C0) and at days in culture C1, C3, C5 and C7. Glomus cells were identified by PNA binding (peanut agglutinin) and absence of response to extracellular ATP. Compared to C0: 1) [Ca2+]i responses to 20mM K+ were not reduced at C1, but were significantly reduced at C3 and C7. 2) [Ca2+]i responses to hypoxia and anoxia in glomus cells were significantly reduced at C1, reduced further at C3 and remained low at C7. 3) Non‐glomus cells (negative PNA labeling, positive ATP response), which initially showed no response to anoxia on C0, developed a [Ca2+]i response to anoxia by C1. 4) [Ca2+]i responses to TEA+4‐AP in glomus cells were not changed during whole 7 day culture period, while the [Ca2+]i response to TEA+4‐AP in non‐glomus cells was significantly increased at C3 and remained so until C7. We conclude that both glomus and non‐glomus cells change their responsiveness in short‐term culture. In particular there is a dramatic decrease in the [Ca2+]i response to hypoxia of glomus cells and development of a [Ca2+]i response in non‐glomus cells to both anoxia and K+ channel blockers.Grant Funding Source: NIH RO1 HL54621
Implementation of rapid prototyping to produce a 3‐dimensional physical model of the anatomical distribution of functionally identified cat pre‐Bötzinger complex microinjection sites (714.4)
Authors: Geritano Mariah , Lam Sharon , O'Neal Marvin , Cordovez Juan , Solomon Irene 
Published: June 2021

Previous work from our laboratory has used histological analyses to confirm functionally identified sites in the cat pre‐Botzinger complex (pre‐BotC). These sites have been represented in a 2‐D fashion based on our histological sections in conjunction with plates from Berman's stereotactic atlas (1968). To gain further insight into the anatomical distribution of these sites, we have used rapid prototyping technology to generate a 3‐D physical model representing the relevant regions/nuclei of the cat brainstem (based on Berman’s atlas) and the histological/stereotaxic coordinates associated with our microinjection sites. To accomplish this, plates from Berman’s atlas were scanned and compiled into a 3‐D configuration slide box, the stereotaxic coordinates of 348 pre‐BötC microinjection sites were entered into an x/y/z matrix based on histologically confirmed coordinates with the origin (0, 0, 0) referenced to the caudal pole of the retrofacial nucleus, midline, and dorsal surface of the medulla, and a 3‐D digital representation was generated and printed. We believe that this approach compliments both the 2‐D representations (previously used) and the digital 3‐D representation, and provides additional insight into the anatomical specificity of our functionally identified and histologically confirmed microinjection sites.Grant Funding Source: Supported by HL 63175