bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Post‐Translational Modifications Related to Cardiovascular Disease
Authors: McComb Mark E , Whelan Stephen A , Spencer Jean L , Heckendorf Christian F , Bachschmid Markus M , Siwik Deborah A , Colucci Wilson , Cohen Richard A , Costello Catherine E 
Published: June 2021

Unfavorable metabolic conditions (metabolic disorders) associated with obesity and diabetes are major causes for cardiovascular disease. Early detection of the adverse effects of metabolic disease remain elusive. We believe that nonspecific changes which occur in plasma proteins, indicators of inflammation and oxidants, may act as evidence of systemic metabolic disease. Here we elucidate potential biomarkers of CVD including both protein changes and changes in post‐translational modifications (PTMs). Label‐free mass spectrometry‐based proteomics was used to interrogate changes in differential protein and PTM expression in plasma samples from mouse and human models. Label‐free LCMS/MS typically yielded >;1,000 features ( p<0.05, >;2fold). A number of cardiovascular disease related proteins were observed. Up regulated proteins were: haptoglobin, a known biomarker related to inflammation, low mannose binding protein associated with inflammation and CVD in type 2 diabetes, superoxide dismutase and extracellular matrix protein both implicated in type 2 diabetes. We also observed different PTMs associated with oxidative stress including lipid peroxidation products such as hydroxynonenal and multiple forms of oxidation such as cysteine sulfonic acid. Development of a metabolic disorder/CVD‐specific protein panel will afford the first step in biomarker panel development such that disease diagnosis and progression may be performed directly at the molecular level.This project was funded by NIH‐NCRR grants P41 RR010888/ GM104603, S10 RR015942, S10 RR020946, S10 RR025082 and NIH‐NHLBI contract N01 HV00239.
Cadherin peptide‐induced enhancement of blood brain barrier (BBB) permeability
Authors: On Ngoc Hong , Laksitorini Marlyn , Kiptoo Paul K , Siahaan Teruna J , Miller Donald W 
Published: June 2021

ObjectiveTo examine the effects of two cadherin peptides, a short linear peptide containing His‐Ala‐Val (HAV) sequence and a cyclic peptide containing the Ala‐Asp‐Thr (cADT) sequence, on BBB integrity.MethodsThe effects of the HAV and cADT peptides on BBB permeability were examined in Balb/c mice. Blood‐brain barrier permeability was assessed with gadolinium contrast agent (Gad; a small hydrophilic permeability marker), IRdye 800cw PEG (a large hydrophilic permeability marker), and Rhodamine 800 (a P‐gp substrate) under control conditions and following exposure to HAV (1.0–32 mM/kg) or cADT peptides (1mM/kg) using magnetic resonance imaging (MRI) and near infrared fluorescence imaging.ResultsMice treated with HAV displayed a dose‐dependent increase in BBB permeability as assessed with Gad enhanced MRI with doses of 1.0 mM/kg having no effect and maximal increases (18‐fold) observed with 32 mM/kg. Increases in BBB permeability were also observed with IRdye 800cw PEG (3‐fold) and Rhodamine 800 (60% increase). The increase in permeability was transient (return to normal within 1 hr). Increases in BBB permeability were also observed with cADT, albeit at lower dose (1.0 mM/kg).ConclusionsCadherin peptides produced a rapid and reversible increase in BBB permeability and the combination of these peptides with therapeutic agents can enhance drug delivery to the brain.Funding:Supported by NSERC and NIH.
Resolving inflammation in stroke through FPR2/ALX
Authors: Gavins Felicity N. E. , Smith Helen K. 
Published: June 2021

Members of the formyl peptide receptor family (in particular FPR2/ALX; Fpr2/3 in mouse) act as endogenous anti‐inflammatory/ pro‐resolving agents. We investigated the pharmacological potential of FPR ligands in modulating inflammation in stroke.Mice underwent bilateral common carotid artery occlusion (5min) and reperfusion (40min or 2h). FPR agonists (Annexin A1 peptide Ac2–26 (AnxA1Ac2–26) and 15‐epi‐lipoxin A4 (15‐epi‐LXA4)) were administered at the start of reperfusion, and resulting cerebral leukocyte‐endothelial (L‐E) interactions were viewed using intravital microscopy.At 40 min reperfusion, 15‐epi‐LXA4 (0.5μg and 4μg/mouse) reduced inflammation vs. vehicle treatment, although this reduction was not sustained at 2h with 0.5μg/mouse. AnxA1Ac2–26 treatment decreased leukocyte adhesion at 40min and 2h vs. vehicle. Combined treatment of AnxA1Ac2–26 and FPR antagonists Boc2 or WRW4 (FPR2/ALX‐selective) abrogated the effects of AnxA1Ac2–26 at 40min, suggesting an Fpr2/3 mechanism. Abrogation was not sustained at to the same extent at 2h.To conclude, 15‐epi‐LXA4 and AnxA1Ac2–26 reduced L‐E interactions in stroke, in a dose and time‐dependent manner, and likely via FPR2/ALX. These novel data indicate that that modulation of FPR2/ALX could produce significant protective effects in clinical stroke.Funded by the BBSRC.
BK channels are not involved in the ryanodine‐induced vasoconstriction of the spiral modiolar artery
Authors: Krishnamoorthy Gayathri , Reimann Katrin , Wangemann Philine 
Published: June 2021

We investigated whether ryanodine‐sensitive Ca2+‐sparks activate BK channels to lower smooth muscle global Ca2+ and cause vasodilation in the spiral modiolar artery (SMA). Ca2+‐sparks in the smooth muscle cells of the SMA were abolished by ryanodine at pressures of 40 and 60 cmH2O. At 40 cmH2O, ryanodine, but not tetracaine, abolished Ca2+‐waves, increased global Ca2+ and constricted SMA. Iberiotoxin, a BK channel inhibitor, had no effect on either global Ca2+ or vascular diameter. However, iberiotoxin increased global Ca2+ and caused a vasoconstriction in the presence of apamin, an inhibitor of SK channels. In contrast to effects at 40 cmH2O, ryanodine did not constrict the SMA at 60 cmH2O. The pressure‐dependence of the ryanodine‐induced vasoconstriction was consistent with the pressure‐dependence of the Ca2+‐sensitivity, which was greater at 40 than at 60 cmH2O. Enhancement of the Ca2+‐sensitivity with 1 nM endothelin‐1 enabled ryanodine‐induced vasoconstriction at 60 cmH2O. These results suggest that BK channels are present but not activated by Ca2+‐sparks in the SMA. Instead, regulation of global Ca2+ and vascular diameter by BK channels is modulated by SK channels. Ryanodine‐induced vasoconstriction is regulated by the Ca2+ sensitivity and may be caused by store‐dependent mechanisms.Supported by NIH‐R01‐DC04280.
Excessive vascular smooth muscle and macrophage proliferation underlies impaired coronary collateral growth in the metabolic syndrome
Authors: Hutcheson Rebecca Lynn , Chaplin Jennifer , Smith Erika , Jadhav Rashmi , Russell James C , Rocic Petra 
Published: June 2021

RationaleCoronary collateral growth (CCG) is impaired in the metabolic syndrome. It is thought that inadequate cell proliferation is a causative factor. Pro‐angiogenic growth factors (GFs) stimulate cell proliferation. Metabolic syndrome patients and animal models exhibit elevated GF levels, but administration of GFs does not promote CCG in the metabolic syndrome.ObjectiveTo determine whether temporally inappropriate excessive cell proliferation underlies impaired CCG in the metabolic syndrome.Methods and ResultsNormal (SD) and metabolic syndrome (JCR) rats underwent transient, repetitive coronary artery occlusion (RI). We have previously shown that CCG was maximal at day 9 of RI in SD rats but did not occur in JCR rats. Cell proliferation was evaluated by immunohistochemistry (PCNA, Ki‐67) and Western blotting (PCNA). The increase in cell proliferation was transient in SD but greater and sustained in JCR rats. Assessment of cell cycle progression confirmed early and transient cell proliferation in SD vs. sustained proliferation in JCR rats. This was associated with accumulation of proliferating cells in the lumen of small arterioles in JCR rats, which failed to undergo outward expansion.ConclusionsExcessive cell proliferation in the later stages of collateral remodeling underlies impaired CCG in the metabolic syndrome. R01 HL093052
Exercise training and hypocaloric diet improves sympathetic arterial baroreflex control in patients with metabolic syndrome and obstructive sleep apnea
Authors: Toschi‐Dias Edgar , Trombetta Ivani C , Silva Valdo J D , Maki‐Nunes Cristiane , Cepeda Felipe X , Alves Maria Janieire N N , Carvalho Glauce , Drager Luciano , Lorenzi‐Filho Geraldo , Negrao Carlos E , Rondon Maria Urbana P B 
Published: June 2021

Metabolic syndrome (MetS) decreases arterial baroreflex control of muscle sympathetic nerve activity (ABRMSNA). And, the obstructive sleep apnea (OSA) exacerbates this autonomic dysfunction. We tested the hypothesis that exercise training and hypocaloric diet (ET+D) would restore ABRMSNA in patients with MetS and OSA. Forty‐four MetS patients were allocated into four groups: Sedentary with OSA (MetS+OSA Sed, n=10) or without OSA (MetS‐OSA Sed, n=10), and ET+D with OSA (MetS+OSA ET+D, n=11) or without OSA (MetS‐OSA ET+D, n=13). The ET+D groups were submitted to −500 kcal/day and 40 min bicycle exercise for 4‐months. OSA was deferred as apnea‐hypopnea index>;15 events/hour. MSNA (microneurography), blood pressure (beat‐to‐beat, noninvasive) and spontaneous ABRMSNA (gain, sensitivity and time delay, latency) were evaluated. ET+D decreased MSNA (P<0.05) and increased ABRMSNA gain in both MetS+OSA (13±1 vs. 24±2 ms/mmHg, P=0.01) and MetS‐OSA (27±3 vs. 37±3ms/mmHg, P=0.03) groups. The time delay of ABRMSNA was reduced by ET+D only in MetS+OSA group (4.1±0.2 vs. 2.8±0.3 s, P=0.04). No changes were observed in the sedentary groups. In conclusion, the ET+D improve ABRMSNA sensitivity in patients with MetS regardless of OSA. However, this effect of ET+D is more pronounced in patients with MetS+OSA.Support: CNPq and FAPESP#2011/17533–6
RAS Blockade Inhibits Bcl‐2 Modifying Factor (Bmf) Expression and Prevents Hypertension and Tubular Apoptosis in Diabetic Akita Mouse Kidneys
Authors: Maachi Hasna , Lo Chao‐Sheng , Abdo Shaaban , Chenier Isabelle , Zhang Shao‐Ling , Chan John S D 
Published: June 2021

BackgroundWe reported that pro‐apoptotic protein Bmf is highly expressed in diabetic mouse renal proximal tubular cells (RPTCs) and induces RPTC apoptosis in vitro (Diabetes 2012). The regulation of Bmf expression in RPTCs, however, is not well understood. We investigated whether renin‐angiotensin system (RAS) blockade could prevent Bmf expression, hypertension and RPTC apoptosis in Akita mice (a model of type 1 diabetes) in vivo and in vitro.MethodsAdult male Akita mice (11 weeks) were treated ± RAS blockers (losartan, 30 mg.Kg−1.day−1 and perindopril, 4 mg.Kg− 1.day−1 in drinking water) until 16 weeks. Controls were untreated non‐Akita littermates. Plasma glucose, systolic blood pressure (SBP) and albuminuria were monitored weekly. Kidneys were processed for histology and apoptosis studies (TUNEL assay). RPTC gene and protein expression were quantified by real time‐qPCR and Western blotting, respectively. We also examined the effect of angiotensin II (Ang II) on Bmf gene expression in cultured rat immortalized RPTCs.ResultsAkita mice developed hyperglycemia and significantly higher SBPs, kidney/body weight ratios and albuminuria, cf. to non‐Akita littermates. Kidneys from Akita mice displayed renal hypertrophy and tubular apoptosis cf. to non‐Akita littermates. Increased expression of Agt, Bmf and active caspase‐3 were evident in RPTCs of Akita mice. Treatment with RAS blockers inhibited Agt and Bmf expression, prevented hypertension and albuminuria and attenuated tubular apoptosis in Akita mice. Finally, Ang II stimulated Bmf mRNA expression in RPTCs and its effect was abolished in the presence of losartan.ConclusionAng II up‐regulates Bmf expression and RAS blockade effectively inhibits Bmf expression and prevents hypertension and tubular apoptosis in diabetic kidneys.Supported by Canadian Institutes of Health Research.
Do exercise and estrogens interact to alter stress responses in rats?
Authors: Jones Alexis Brooke , Gupton Rebecca , Curtis Kathleen S 
Published: June 2021

The beneficial effects of exercise in coping with stress/anxiety may be particularly important for women, as anxiety and depression increase after menopause. The present experiments examined anxiety‐like behaviors and corticosterone (CORT) levels induced by acute stress in female rats with or without estradiol benzoate (EB) after a single bout of exercise vs. 3 days of exercise before and after stress.Adult female Sprague Dawley rats were ovariectomized, recovered for 7 days, given EB or OIL for 2 consecutive days, and tested during the next 2 days. In Expt. 1, rats were stressed by being placed in a plastic restrainer for 40 min, then were returned to a novel cage with a running wheel attached. Software recorded distance, speed, and time spent on wheels for 14 hrs. The next morning, anxiety‐like behaviors were assessed using the Elevated Plus Maze (EPM). In Expt. 2, exercising rats had access to running wheels beginning on day 1 of EB/OIL treatment and ran throughout the experiment, including the 14 hrs after restraint stress.Exercise prior to restraint stress reduced CORT levels in EB‐treated rats compared to rats that ran after stress. However, 3 days of exercise increased anxiety‐like behaviors in both groups compared to rats exercised for only 1 day, with or without restraint stress.Thus, exercise duration affects stress hormones in EB‐treated rats, but CORT levels may not accurately predict anxiety‐like behaviors.
Intermittent respiratory stimulation with doxapram induces phrenic motor plasticity
Authors: Sandhu Milap Singh , Lee Kun‐Ze , Reier Paul J , Fuller David D 
Published: June 2021

Intermittent hypoxia can induce a persistent increase in phrenic motor output via a central neural mechanism. Here we explored a pharmacological alterative to hypoxia as a tool for triggering phrenic motor plasticity. Specifically, we hypothesized that persistent increases in efferent phrenic burst activity can be evoked via systemic delivery of a low‐dose of doxapram – a drug which can robustly stimulate phrenic activity due to activation of carotid chemoafferents. Anesthetized, vagotomized, paralyzed and mechanically‐ventilated rats were given intravenous doxapram either intermittently (3 × 2mg/kg) or as a single dose (2mg/kg or 6mg/kg). A persistent increase in phrenic burst amplitude was evoked by intermittent doxapram (161 ± 27% baseline at 60 min post‐injection, p<0.05 vs. sham injection and time control), but not by the single dose (104 ± 13% baseline). The acute phrenic response was substantially reduced in carotid‐denervated rats (120 ± 3% baseline compared to 165 ± 9% baseline in animals with intact carotid nerves, p<0.05) confirming that doxapram acts primarily via the carotid chemoreceptors. Since doxapram is an FDA approved drug and commonly used as a respiratory stimulant, it may be a viable alternative or supplement to hypoxia for targeted induction of respiratory neuroplasticity in neurological conditions such as spinal cord injury.Support: Craig H. Neilsen Foundation #220521 (MSS), 1R01NS080180–01A1 (DDF), NIH1R21HL104294–01 (DDF)
Ouabain increases association between Na‐K ATPase (Na‐K) and NHE1 through N‐terminal domain of Na‐K
Authors: Khundmiri Syed J , Xie Zijian , Lederer Eleanor D 
Published: June 2021

Low dose ouabain increases Na‐K activity in kidney basolateral membranes in an NHE1 dependent manner. TIRF and FRET microscopy suggested that 10 pM ouabain increases association between Na‐K and NHE1. The aim of the study was to identify the molecular domains on Na‐K and NHE1 required for association. Using mouse (MRPT) and human (HK11) kidney proximal tubule cells co‐transfected with full‐length GFP or RFP‐NHE1, and full length mCherry‐Na‐K a‐subunit or N‐terminal deletion mutant of YFP‐Na‐K, FRET and TIRFM were performed before and after ouabain treatment. We also measured the effect of ouabain on Na‐K activity in HK2 cells transfected with WT or scaffold domain mutated NHE1. Ouabain increased association between Na‐K and NHE1. NHE1 and Na‐K association was abolished in cells expressing N‐terminal deletion mutant Na‐K. Ouabain increased Na‐K activity in HK2 cells expressing full length NHE1 but not in cells expressing scaffold domain mutated NHE1. These data suggest that low concentrations of ouabain increase Na‐K association with NHE1 and this association involves N‐terminal domain of Na‐K and the scaffolding domain of NHE1.Supported by AHA and VA
Quantification of tendon cell nuclei in rat Achilles tendon after 12 weeks of running
Authors: Couppé Christian , Thomsen Emilie Wøjdemann , Svensson René , Heinemeier Katja , Kongsgaard Mads , Kjaer Michael , Magnusson S. Peter , Scherling Peter 
Published: June 2021

IntroductionIt remains unknown why tendons improve mechanical properties with loading. However, tendon cell nuclei (TCN) have been claimed to induce these mechanical adaptations, although this has not been previously investigated.PurposeTo quantify the number of TCN in rat Achilles tendon after 12 wk of running.Methods8 adult male Sprague‐Dawley rats ran (RR) on a treadmill with 10° incline, 1 h/day, 5 days/wk (17–20 m/min) for 12 wk (which led to improved tendon mechanical properties) and were compared with 11 control rats (CR). Tissue‐Tek embedded serial sections (10 μm) from the mid region of the Achilles tendon were cut longitudinally on a cryostat. Sections were stained with PicroSirius Red and hematoxylin staining. One investigator counted the number of TCN 2–3 times in three separate parts of the mid longitudinal tendon section with fields of 390 μm ×280 μm in a blinded fashion. Unpaired t‐test was used for the statistical analysis (Mean±SE).ResultsTypical Error % for the three parts was 5.3%: There was no difference between running rats vs. control rats (TCN per image (≈ 105μm): RR, 152±9; CR, 146±8, P=0.642).ConclusionThis new method provided reproducible determination of tendon cell nuclei density. However, even though the tendon mechanical properties were improved, no difference in tendon cell nuclei density between running rats vs. control rats could be detected. Research support: Danish Association of Rheumatism.
Function and molecular regulation of WNT‐5A expression by TGF‐β
Authors: Kumawat Kuldeep , Menzen Mark H , Halayko Andrew J , Schmidt Martina , Gosens Reinoud 
Published: June 2021

Disruption of WNT‐5A homeostasis has been linked to various disorders like fibrosis, cancer and inflammation with recent evidence implicating it in asthma. Here, we have investigated the function and molecular regulation of WNT‐5A expression in airway smooth muscle (ASM) cells by TGF‐β.Immortalized human ASM cells were used to measure mRNA by qRT‐PCR and protein abundance by immunoblotting.ASM cells expressed a variety of WNT ligands and Frizzled receptors which were differentially modulated by TGF‐β. WNT‐5A was the most abundant ligand expressed and showed ~4–16 fold upregulation in response to TGF‐β (2 ng/mL). Knockdown using siRNA demonstrated WNT‐5A requirement for TGF‐β‐induced collagen and fibronectin expression. TGF‐β induction of WNT‐5A appeared to require TGF‐β activated kinase (TAK)‐1 activity, as pharmacological inhibition of TAK‐1 with LL‐Z1640–2 (500 nM) prevented WNT5A expression. Interestingly, inhibition of ERK1/2 (U0126, 3μM), Smad3 (SIS3, 3μM) and β‐catenin/TCF (PKF115–584, 100 nM) augmented WNT‐5A expression suggesting a negative regulatory role.In conclusion, TGF‐β‐induced WNT‐5A expression mediates extracellular matrix expression by human ASM cells. WNT5A induction occurs via complex signaling cascades with TAK‐1 acting as a positive regulator, whereas ERK1/2, Smad3 and β‐catenin/TCF mediate a negative constraint.This study is funded by University of Groningen.
Small litter size at birth is associated with insulin resistance and low energy expenditure in adulthood
Authors: Tavares Neuziane Kloos Amorim , de Jesus Aguiar Fernanda , Furukawa Luzia Naôko , Heimann Joel Claudio 
Published: June 2021

ObjectiveTo evaluate the effect of litter size standardization at birth on adult metabolic parameters.MethodsFemale Wistar rats became pregnant at 12 weeks of age. Offspring were divided into control(C), G8(8) and G4(4 newborns/litter) groups. At 12 weeks of age, food intake (FI,g/day), glucose (G,mg/dL), insulin (I,pM), cholesterol (Chol,mg/dL), triacylglycerols (TAG,mg/dL) levels, white and brown adipose tissue (BAT) masses (g/100g) adiposity index (AI,% body weight) were determined.Results(mean±SEM p<0.05, n=6 to 32):

Male
C
G8
G4

AI
4±0.1
4±0.1
5±0.51

FI
26±0.7
24±0.9
17±21

BAT
0.13±0.01
0.11±0.003
0.09±0.011

G
109±3
100±3
120±41

I
129±13
126±10
203±361

Chol
81±32
66±3
69±5

TAG
50±4
67±53
67±43

BP
127±44
101±6
121±124

p<0.05 vs C and G8;
p<0.05 vs G8 and G4;
p<0.05 vs C;
p<0.05 vs G8

Female
C
G8
G4

AI
5±0.3
5±0.3
7±0.71

FI
17±0.82
15±0.8
16.6±0.92

BAT
0.16±0.022
0.13±0.007
0.11±0.0051

G
92±42
89±2
104±41

I
50±62
71±9
106±161

Chol
57±3
62±4
66±53

TAG
46±5
63±53
65±73

BP
125±74
99±6
106±82

p<0.05 vs C and G8;
p<0.05 vs G8;
p<0.05 vs C;
p<0.05 vs G8 and SG4

ConclusionResults suggest that changes in litter size is associated with energy balance, carbohydrate and lipid disorders, insulin resistance and alterations in blood pressure in adult male and female.Supported by FAPESP
Targeted expression of SLC26A4 rescues hearing and balance in Slc26a4Δ/Δ mice
Authors: Wangemann Philine , Li Xiangming , Sanneman Joel , Zhou Fei , Harbidge Donald G. , Nelson Raoul D. , Chambrey Régine , Eladari Dominique , Miesner Tracy , Marcus Daniel C. 
Published: June 2021

Mutations of SLC26A4 are a common cause for deafness associated with enlarged vestibular aqueduct. SLC26A4 encodes pendrin, an anion exchanger expressed in a variety of epithelial cells in the cochlea and the vestibular labyrinth. Slc26a4Δ/Δ mice develop a severe enlargement of the membranous labyrinth, fail to acquire hearing and balance, and thereby provide a stark model for the human phenotype. Here we generated a transgenic mouse line that expresses SLC26A4 under the promoter of ATP6V1B1. Crossing this transgene into the Slc26a4Δ/Δ line restored the temporal and spatial expression pattern of pendrin in the endolymphatic sac without inducing expression in the cochlea or the vestibular sensory organs. The transgene prevented enlargement and restored normal hearing and balance. Our study demonstrates that a limited restoration of pendrin is sufficient to restore normal inner ear function. These data in conjunction with notion that pendrin is required for the development but not the maintenance of hearing (Choi et al., 2011) opens the prospect that a spatially and temporally limited therapy may restore a life‐time of normal hearing in Slc26a4Δ/Δ mice, and possibly humans with SLC26A4 mutations.Supported by NIH‐R01‐DC012151, NIH‐P20‐RR 17686, KINBRE, Kansas State University, ANR BLANC 2010‐R10164DD