bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age‐Related Left Ventricular Systolic Dysfunction in a Mouse Model of Hypertrophic Cardiomyopathy
Authors: Larson Robert A. , Purnell Benton S. , Davis Melissa K. , Weiss Robert M. , Cicha Michael Z. , Sabharwal Rasna , Chapleau Mark W. 
Published: November 2023

Familial hypertrophic cardiomyopathy (HCM) is a devastating disease caused by mutations in cardiac sarcomeric proteins that result in myocyte disarray, cardiac fibrosis and hypertrophy. A hallmark of HCM is cardiac diastolic dysfunction. Systolic function is generally preserved or even enhanced; however, some patients develop systolic dysfunction over time. In this study we tested the hypothesis that left ventricular (LV) systolic dysfunction increases with age in transgenic mice with cardiac‐specific expression of mutated alpha tropomyosin (Glu180Gly), an established model of human HCM. Cardiac function was assessed by echocardiography (Vevo 2100, Visualsonics) in HCM and control wild type (WT) mice over a wide range of age (3–47 weeks). Mice were lightly sedated with midazolam (0.15 mg, SC) to avoid anesthesia‐induced decreases in heart rate, myocardial contractility and sympathetic tone. LV end diastolic volume (EDV) was not significantly different in HCM vs. WT mice at any age (see Table). LV end systolic volume (ESV) and ejection fraction (EF) did not differ between very young, 3‐week old WT and HCM mice. In contrast, at 7 weeks of age, ESV was higher and EF was lower in HCM vs. WT mice (P<0.05). Systolic dysfunction was most pronounced in middle‐aged HCM mice between 36 and 47 weeks of age (Table, P<0.05). We conclude that alpha tropomyosin mutant mice with HCM exhibit age‐dependent LV systolic dysfunction.Support or Funding InformationAAS‐Lundbeck Fellowship & HL14388

3 Weeks
7 Weeks
43±1 Weeks

WT (n=8)
HCM (n=9)
WT (n=8)
HCM (n=9)
WT (n=5)
HCM (n=12)

EDV (μL)
24.2±2.4
28.8±2.0
37.4±4.8
45.4±4.3
41.6±7.3
54.0±3.7

ESV (μL)
6.7±1.1
9.5±0.9
9.1+1.5
18.0±2.8 *
10.1±2.0
21.3±1.7*

EF (%)
73±2
67±1
76±2
62±3*
76±3
61±2*

p<0.05 WT vs. HCM
Depressed State of Cardiac Energetics Following Cardiopulmonary Bypass in Pigs
Authors: Jaimes Rafael , Sarkislali Kamil , Sherman Meredith , Swift Luther , Ishibashi Nobuyuki , Posnack Nikki 
Published: November 2023

ObjectiveThe objective of this work is to determine the state of cardiac energetics after hypoxia due to cross‐clamping of the aorta during cardiopulmonary bypass in an open‐heart, pig model.MethodsBlood samples and apical, left‐ventricular biopsies (~1g) were taken after 2 hours of cardiopulmonary bypass. Blood samples were measured for lactate dehydrogenase (LDH) activity. The key modulating enzyme for glucose oxidation, pyruvate dehydrogenase (PDH), was assessed for activity using isolated mitochondria from the myocardium. Biopsies were incubated with sodium fluoride and phenylmethylsulfonyl fluoride, then homogenized and underwent a series of centrifugation to isolate mitochondria. Mitochondrial membranes were permeated using Triton‐X and then mixed with PDH substrates (thiamine tri‐phosphate, MgCl2, NAD+, CoA, and pyruvate). PDH kinetics were assessed by monitoring NADH absorbance at 340nm with a spectrophotometer. The sub‐maximal PDH activity was determined by using 2mM pyruvate and maximal activity was determined with 20mM pyruvate. The ratio of sub‐maximal to maximal activity provided the activated fraction.Summary of ResultsPDH was found to be 42.3% ± 7.8 activated in bypass pigs (n=3), compared to 71% ± 12.4 activated in control pigs (n=2). Lactate dehydrogenase level was 444±123 IU/L in bypass pigs compared to the reference range of 105–333 IU/L.ConclusionPDH is inactivated by the buildup of the electron carrier NADH and ATP, which occurs in hypoxic conditions. Lack of post‐ischemic, re‐activation of PDH results in an accumulation of pyruvate and activation of LDH, which is associated with decreased functional recovery (previously shown in isolated rodent hearts). This work shows that aortic‐cross clamping causes inactivation of PDH due to hypoxia. Reduced PDH activity can inhibit electrical and mechanical cardiac performance during the vulnerable post‐operative period.Support or Funding InformationThis study was supported by the National Institutes of Health (R00ES023477 to NGP, UL1TR000075, KL2TR000076), Children's Research Institute: Divisions of Cardiology and Cardiac Surgery.
A novel cross talk between mechanosensitive ion channel TRPV4 and VEGFR2 in human endothelial cells
Authors: Kanugula Anantha Koteswararao , Adapala Ravi Kumar , Cappelli Holly C , Paruchrui Sailaja , Thodeti Charles K 
Published: November 2023

Angiogenesis is critical for cardiovascular function, however, insufficient or excessive angiogenesis has been implicated in various diseases. Although both soluble (VEGF) and mechanical factors regulate angiogenesis, most of the studies on angiogenesis are focused on targeting soluble signaling. We have recently shown that mechanosensitive ion channel TRPV4 (transient receptor potential vanilloid 4) negatively regulates angiogenesis via modulation of Rho/Rho kinase‐dependent endothelial mechanosensing. We also found that pharmacological activation of TRPV4 normalized abnormal tumor vasculature and improved cancer therapy. However, it is not known if there is a cross‐talk exists between TRPV4 and VEGF signaling. To explore this cross‐talk, we knocked down TRPV4 in human endothelial cells and measured VEGFR2 expression, localization and phosphorylation. We found a significant increase in VEGFR2 phosphorylation at Y951 (kinase insert domain) and Y1175 (c‐terminal domain) in TRPV4 siRNA knocked down cells. Further, we found that knockdown of TRPV4 increased VEGFR2 expression with concomitant increase in angiogenic transcriptional factor GATA2. Importantly, immunofluorescence analysis revealed reduced accumulation of VEGFR2 in perinuclear Golgi compartment suggesting increased trafficking of VEGFR2. Taken together, our results suggest that TRPV4 mechanotransduction regulates VEGFR2 expression and trafficking and identifies a novel cross‐talk between mechanical (TRPV4) and soluble (VEGF) signaling in human endothelial cells.Support or Funding InformationNIH‐NHLBI (R01), NIH‐NCI (R15), AHA (Grant‐in‐Aid), NIH‐NHLBI (R15)
SMAD3 contributes to lung vascular remodeling in pulmonary arterial hypertension via MRTF disinhibition
Authors: Zabini Diana , Granton Elise , Bonnet Sandra Breuils , Bonnet Sebastién , Morrell Nicholas , Connelly Kim , Provencher Steeve , Ghanim Bahil , Klepetko Walter , Olschewski Andrea , Kapus Andras , Kuebler Wolfgang 
Published: November 2023

IntroductionVascular remodeling in pulmonary arterial hypertension (PAH) results from smooth muscle cell hypertrophy and proliferation of vascular cells. Loss of bone morphogenetic protein receptor 2 (BMPR‐II) signaling and increased signaling via transforming growth factor β (TGF‐β) and its downstream mediators SMAD2/3 has been proposed to drive lung vascular remodeling; yet, proteomic analyses indicate a loss of SMAD3 in PAH.ObjectiveWe thus proposed that SMAD3 may be dysregulated in PAH, and that loss of SMAD3 may present a master switch in the disease process. Specifically we hypothesized that SMAD3 loss may promote lung vascular remodeling via disinhibition of its interaction partner myocardin‐related transcription factor (MRTF) which drives muscle protein expression.Methods and ResultsSMAD3 was downregulated in lungs of PAH patients, and in pulmonary arteries of three PAH animal models (monocrotaline or sugen/hypoxia rats, and BMPR2+/R899X mice with dysfunctional BMPR‐II). TGF‐β treatment replicated the loss of SMAD3 in pulmonary artery smooth muscle (huPASMCs) and endothelial (huPAECs) cells. SMAD3 silencing increased proliferation and migration in huPASMCs and huPAECs. Co‐immunoprecipitation revealed reduced interaction of MRTF with SMAD3 in TGF‐β treated huPASMCs and pulmonary arteries of PAH animal models. In huPASMC, loss of SMAD3 or BMPR‐II increased smooth muscle actin expression, which was attenuated by MRTF inhibition. Conversely, SMAD3 overexpression prevented actin stress fiber formation in BMPR2 silenced huPASMC. MRTF inhibition attenuated PAH and lung vascular remodeling in sugen/hypoxia rats.ConclusionLoss of SMAD3 presents a novel central pathomechanism in PAH that promotes both vascular cell proliferation and ‐ via MRTF disinhibition ‐ hypertrophy of huPASMC, thereby reconciling the parallel induction of a synthetic and contractile huPASMC phenotype. While basal MRTF signaling has been shown to maintain BMP‐regulated vascular homeostasis in healthy lungs, disinhibition of MRTF following functional loss of BMPR‐II and subsequent downregulation of SMAD3 drives lung vascular remodeling in PAH.Support or Funding InformationThis work was supported by the Erwin‐Schroedinger Fellowship of the Austrian FWF Foundation to DZ, and grants‐in‐aid from the Canadian Institutes of Health Research (CIHR) and the Heart & Stroke Foundation of Canada (HSFC) to WMK
Rapid High‐Fat Diet‐Induced Glucose Intolerance is Not Associated with Hepatic Interleukin‐6 Resistance
Authors: Townsend Logan K. , Trottier Sarah K. , Peppler Will T. , Wright David C. 
Published: November 2023

BackgroundHigh‐fat diet (HFD)‐induced hepatic insulin resistance is associated with attenuated hepatic interleukin‐6 (IL‐6) action. This is important as ablation of hepatic IL‐6 signalling worsens systemic glucose tolerance. Hepatic IL‐6 resistance may result from increased content or activity of Src homology phosphatase 2 (SHP2) in the liver, a phosphatase which dephosphorylates STAT3, a downstream IL‐6 signaling intermediate.ObjectiveDetermine if the initial development of glucose intolerance with a high fat diet is associated with the onset of liver IL‐6 resistance.HypothesisHFD will rapidly induce hepatic IL‐6 resistance, in association with impairments in whole body glucose homeostasis and increased liver SHP2 content and activity.MethodsC57bl/6 mice (10 per group) had ad libitum access to standard chow or HFD (60% kcal from fat) for 3 d. On the 4th d, mice were fasted for 6 h before an intraperitoneal (IP) glucose tolerance test. The following day half of the mice in each group were injected IP with IL‐6 (3 ηg kg−1 body weight) or sterile saline. Tissues were harvested 15 min post‐injection. STAT3 phosphorylation and SHP2 activity were assessed via Western blotting.ResultsMice fed a HFD were glucose intolerant (P<0.001). This was not associated with attenuated hepatic STAT‐3 phosphorylation following IL‐6 injection. Nor was hepatic SHP2 content or activity increased in HFD mice.ConclusionsHFD rapidly causes systemic glucose intolerance but this is not the result of blunted hepatic IL‐6 signalling.Support or Funding InformationOntario Graduate Scholarship and Dairy Farmers of Ontario Doctoral Research Assistantship (LKT) and Natural Sciences and Engineering Research Council and Canada Research Chair (DCW)
Lipopolysaccharide Induced Activation of VRAC (LRRC8) in Nodose Neurons Is Dependent on TLR4 Receptor Mediated ROS Generation
Authors: Wang Runping , Lu Yongjun , Cicha Michael Z , Chapleau Mark W , Abboud Francois M 
Published: November 2023

The Volume‐Regulated Anion Channel and its essential subunit Leucine Rich Repeats Containing protein 8a (VRAC/LRRC8) is a widely expressed Cl− channel critical for cell volume regulation. This channel is activated by osmotic gradient and ionic strength (Voss et al., 2014, Qiu et al., 2014, Syeda et al., 2016). Using patch‐clamp techniques, we reported that channel activation is also induced in the majority of vagal afferent neurons isolated from mouse (C57BL/6) nodose ganglia by hypoosmolarity and by extracellular low pH (Wang et al., 2017). We have also observed that this current is activated by lipopolysaccharide (LPS) at 10μg/ml. Here we tested the hypothesis that the response to LPS is dependent on toll‐like receptor 4 (TLR4) and is mediated by intracellular reactive oxygen species (ROS). The current was reduced from 26.7±5.4 pA/pF (n=5) in control peptide‐treated neurons to 6.4±2.1 pA/pF (n=6, p<0.05) in neurons treated with the TLR4 inhibitor peptide VIPER. Application of LPS at a lower dose (100ng/ml) increased intracellular neuronal ROS measured by dihydroethidium within several minutes. The cell permeable hydrogen peroxide scavenger PEG‐catalase decreased the LPS‐induced current from 10.2±2.5 pA/pF (n=7) to 2.3±1.2 pA/pF (n=5, p<0.05). We conclude that LPS induces activation of the VRAC/LRRC8 channel in nodose neurons via TLR4 receptor‐mediated ROS generation. The results indicate a diverse functional role of this channel beyond volume homeostasis with potential implications in high fat diet‐ or LPS‐induced decrease in satiety signaling in intestinal vagal afferents.Support or Funding InformationHL14388
Chemical and Histological Analysis of Leptomeningeal Plaques
Authors: Coovert Daniel , Millspaugh‐Storms Brittany , Fitch Richard 
Published: November 2023

Study ObjectiveThis study was carried out to investigate the composition of arachnoidal plaques discovered during routine cadaver dissection during gross anatomy courses at Indiana State University.BackgroundLeptomeningeal calcified plaques are uncommon formations that have been periodically described in dissection or in autopsy for over 100 years (1,2). These typically benign leptomeningeal plaques are usually asymptomatic, and are distinct from those associated with the chronic proliferative disorder Arachnoiditis ossificans (AO). The etiological cause of arachnoidal plaques and AO are unknown, but iatrogenic, traumatic, metastatic, and infective factors have been associated with these disorders (2).MethodsLeptomeningeal plaques identified during routine cadaver dissection were excised and examined by SDS‐PAGE, histological stains, and electron microscopy. Proteins present in the inclusion were resolved on a 10% gel, and were detected using silver staining. Presence of surface protein was detected using amidoblack. Ultrastructral analysis and elemental composition was carried out using a JOEL 7008F Field Emission Scanning Electron Microscope at the Integrated Nanosystems Development Institute (INDI) at Purdue University Indianapolis.ResultsAmidoblack staining indicated the plaques are covered by a thin protein layer derived from the arachnoid. Electron microscopy confirmed the surface is composed of a fibrous network, presumably collagen fibers derived from the arachnoid. These results indicate the plaque formation process initiates within the arachnoid rather than aggregating on the luminal surface of the arachnoid and growing into the subarachnoid space. Elemental analysis indicates the plaque is highly biased toward carbon with little nitrogen and lacks calcium or phosphorus. This suggests the plaque is highly enriched with lipid. Protein analysis indicated the protein composition was similar to the composition of the arachnoid.ConclusionsLeptomeningeal plaques are lipid rich inclusions that form within the arachnoid layer.
Hypothalamic vasopressinergic and orexinergic pathways converge with midbrain aminergic inputs on LHb intrinsic GABAergic system regulated by neurosteroid signaling: implications for contingency‐dependent motivated behavior
Authors: Zhang Limei , Hernandez Vito S. , Eiden Lee E. 
Published: November 2023

We have recently reported the existence of a vasopressinergic input from the hypothalamic paraventricular nucleus (PVN) to the medial division of the lateral habenula (LHbM), reporting to a subpopulation of putative GABAergic interneurons there. Stimulation of this pathway influences escape behaviors in the rat. Here, we identify additional hunger‐related orexinergic and reward‐related midbrain aminergic (dopaminergic and serotoninergic) inputs converging on this habenular cell group, and uniquely expressing the estrogen receptor ERalpha. The origins of each of the projection types were determined by fluorogold retrograde tracing. Orexinergic, dopaminergic and serotonergic projections originate in lateral hypothalamus (LH), ventral tegmental area (VTA) and substantia nigra (SN), and dorsal raphe (DR), respectively. They share, with the vasopressinergic inputs from the PVN, sex‐steroid responsivity, i.e. they express androgen receptors (ARs) and aromatase, the latter visualizable in axon terminals within the habenula. The distinct LHbM cell population onto which these inputs converge are likely to be functionally GABAergic, as determined by in situ hybridization histochemistry identifying the expression of gad1, gad2, Slc6A1, Slc6A11, Slc32A1 within them. This cell group also expresses mRNA encoding receptors for serotonin, dopamine, vasopressin, and orexin. Of physiological importance, AR and aromatase expression in the input neurons from PVN, LH, VTA/SN and DR are dependent on both estrogen levels, in female rats, and history of sexual activity, in male rats. Castration‐induced behavioral changes in passive vs active coping strategies before a predator were also observed. We postulate that all four pathways studied here contribute to the hormonal/homeostatic control of behavioral motivation in a coordinated or convergent fashion, through the use of estrogen as a co‐transmitter impingent on LHbM neurons. These findings are relevant within a broader physiological context, reflecting the influence of hunger and thirst circuitry on contingency‐dependent motivated behavior.Support or Funding InformationPartially supported by: DGAPA‐UNAM‐PAPIIT‐IN216214, CONACYT‐CB‐176919 & CB‐238744 to LZ and NIMH‐IRP‐1ZIAMH002386 to LEE. LZ is a Fulbright visiting scholar, also supported by PASPA‐DGAPA‐UNAM fellowships for her sabbatical research stay hosted by LEE of SMN‐NIMH‐NIH‐USA.
Quantum Dynamics Neuronal Stimulation by Sigma Photonics Quantum Energy
Authors: Einstein George P , Anaushvili Avtavandil P , Tulp Orien L 
Published: November 2023

It is well known that the human brain generates waves (oscillation). For example, it is known that brain generates electromagnetic waves, and this phenomenon is used in electro encephalography. In a given model of a human brain, the two hemispheres are considered as two independent generators of oscillation processes. The mathematical description of the specified wave model of a brain has been determined (Anuashvili). The mathematical description of the specified wave model of a brainwill be presented. For the oscillatory processes occurring in the hemispheres, we shall present the mathematical formulae we have developed. The domination of one of hemispheres can be determined by the size of oscillitory displacement of the source of the oscillations during an exposition or proportional value ‐ difference of amplitudes of oscillatory processes occurring in hemispheres. Thus, in wave model of a brain two parameters are allocated:

Domination of one of hemispheres,
Coordination (coherence) of oscillatory processes in hemispheres.
Thus, this research determned the mathematical formulae to describe hemispheric electromagnetic wave functions and may contribute to the clinical diagnostic profile of the individuals under study under a variety of clinical conditions.Support or Funding InformationInstitutional Support of the Einstein Medical Institute and the College of Medicine, University of Science Arts and Technology, Montserrat, British SWest Indies.
NMDA receptor 2B Subunit and BK Channel May Affect Mechanical Allodynia by Regulating the Activity of Enkephalinergic Neurons in the Spinal Dorsal Horn
Authors: Tanaka Shiho , Kato Eiko , Kofuji Takefumi , Fukushima Teruyuki , Hori Yuuichi 
Published: November 2023

Enkephalin (ENK)‐containing neurons in the spinal superficial dorsal horn (SDH) play an important role in regulating the transmission of nociceptive information. Using BAC transgenic mice, in which ENK‐containing neurons can be identified by green fluorescent protein expressed under the control of preproenkephalin gene promoter, we have observed that ENK‐containing neurons in the SDH express the NMDA receptor 2B (NR2B) subunit and large conductance calcium‐activated potassium (BK) channels. In experiments, we investigated the effects on nociceptive behavior of intrathecally administrating NR2B subunit antagonist and BK channel antagonist in peripheral nerve‐injured mice.The experiments were performed in 6‐ to 8‐week‐old male ICR mice. Partial sciatic nerve ligation was performed as described previously (PSL, Seltzer model). To assess the development of mechanical allodynia, we measured the frequency of withdrawal responses to 10 repetitive stimuli with von Frey filaments. Behavioral assessments were performed beginning 3 days prior to the ligation surgery. On post‐surgical day 7, effects on nociceptive behavior of the BK channel antagonist, charybdotoxin (CTX), and the NR2B antagonist, ifenprodil (IFN), were analyzed. Additionally, in order to clarify whether the actions of CTX and IFN are mediated by an alteration in the release of enkephalin, we investigated the effect of the selective δ‐opioid receptor antagonist naltrindole (NTI).CTX (1 pmol/10 μL per mouse), IFN (50 nmol/10 μL per mouse) and saline as a control (10 μL per mouse) was intrathecally injected according to the modified method originally described by Hylden. All behavioral experiments were performed in double‐blind fashion.PSL significantly increased the occurrence of withdrawal reflex to von Frey stimuli being indicative of the development of mechanical allodynia. Intrathecal CTX significantly reduced the occurrence of withdrawal reflex as compared to saline group. Additionally, intraperitoneal administration of NTI at the dose of 5 mg/kg body weight attenuated the effect on nociceptive behavior of CTX. Intrathecal IFN significantly reduced the occurrence of withdrawal reflex. This effect was also blocked by intraperitoneal NTI.These behavioral observations might suggest that NMDA receptors and BK channels inhibit the activity of enkephalin‐containing neurons in the spinal dorsal horn, which is speculated to be involved in peripheral nerve injury‐induced chronic pain state.Support or Funding InformationThis work was supported by Dokkyo Medical University Endowment Fund.
Investigating the Role of the DYW Deaminase in RNA Processing
Authors: Aldana‐Mendoza Jonathan A , Hayes Michael lloyd 
Published: November 2023

RNA editing is the process by which ribonucleic acid (RNA) molecules are enzymatically modified post synthesis on specific nucleosides. In land plants it is mostly comprised of cytidine (C) to uridine (U) transformations linked to proteins with the DYW deaminase domain within the pentatricopeptide repeat (PPR) protein family. The DWY deaminase domain is likely the catalytic RNA editing domain because of its conserved glutamate residue of the HXE motif, required in other known catalyzed cytidine deaminases. PPR proteins have been shown to play important roles in transcript maturation. The molecular specificity of RNA‐binding proteins is essential for RNA regulation. Understanding RNA recognition also allows re‐coding of RNA‐binding proteins that may be employed to modulate RNA regulation in vivo. We plan on investigating the binding of PPRs to natural RNA transcripts by using transition state analogs (TSA's) which could inhibit RNA editing in vitro. Using different synthetic RNA's we can identify the requirements for RNA editing activity. Using chloroplast extracts in vitro RNA editing efficiency can be compared to determine the necessary cis‐‐elements. In order to discover the associated PPR specificity factor. This would also provide insight into the degenerate recognition code of RNA editing.Support or Funding InformationNIH Grant # GM 49001