bluuebunny/crossref_metadata_2025_split huggingface dataset:
Calmodulin regulates GPER/GPR30‐mediated signaling
Authors: Matnishian Vahe , Jasurda Jake , Tran Phuong , VerMeer Mark , TRAN QUANG‐KIM 
Published: November 2023

The G protein‐coupled estrogen receptor 1 (GPER/GPR30) was recently identified as a novel receptor for estrogen, and has been demonstrated to be involved in numerous processes in various organ systems. However, its regulatory inputs are not entirely understood. Calmodulin (CaM) is the ubiquitous transducer of Ca2+ signals and is required for the functions of numerous cellular proteins. We recently developed FRET biosensors to identify and characterize four distinct CaM‐binding domains in GPER/GPR30. These domains are located separately in GPER/GPR30's submembrane domains (SMDs) 1–3 and the juxtamembranous (JM) segment of SMD4 (SMD4JM) (Tran et al., PLoS ONE 9(2): e89669). To investigate the impact of CaM binding at these domains on GPER/GPR30‐mediated signaling, we generated a series of mutations in SMD2, SMD3 and SMD4JM to alter hydrophobicity and charges of the original domains. Resultant biosensors developed on the mutant domains, BSGPER150–175mut, BSGPER242–259mut, and BSGPER330–351mut still display classic conformational changes upon CaM binding, yet with substantially reduced affinities. The Ca2+ sensitivities of the interactions between CaM and these domains also are decreased. Heterologous expression of wild‐type GPER/GPR30 increases phosphorylation of the extracellular related kinase (ERK1/2). CaM‐binding reducing mutations in SMD2, SMD3, or SMD4 do not significantly affect pre‐association between GPER/GPR30 and Gbg, yet substantially block GPER/GPR30‐mediated ERK1/2 phosphorylation. These data demonstrate that CaM binding to GPER/GPR30 plays an important role in GPER/GPR30‐mediated signaling.Support or Funding InformationNational Institutes of Health Grant HL112184
Lesphedeza bicolor Ameliorates Methylglyoxal‐induced Dysfunction in Human Aortic Endothelial Cells via Regulation of Oxidative Stress and Apoptotic Signaling Cascades
Authors: Do Moon Ho , Lee Jae Hyuk , Subedi Lalita , Kang Min Cheol , Kim Sun Yeou 
Published: November 2023

Reactive intermediates of glucose, especially methylglyoxal(MGO) can form advanced glycation products, which relate to the diabetic complications. Methylglyoxal is known to cause damage and induce apoptosis in endothelial cells. Endothelial cell apoptosis is implicated in the progression of diabetes complications and atherosclerosis. Lesphedeza bicolor (LB) has been used as a traditional folk medicine for the removal of various toxic materials, the replenishment of energy stores, and for the regulation of diabetes. The effects of LB on MGO‐induced apoptosis were confirmed by annexin V/FITC staining and western blot. Inhibition of LB on MGO‐induced ROS generation was measured using a JuLI live‐cell imaging system. The present study was conducted to investigate the antiglucotoxicity of the stalk of LB on MGO‐induced apoptosis in human aortic endothelial cells (HAoECs). Pretreatment with LB significantly inhibited MGO‐induced cellular apoptosis and generation of intracellular ROS. LB prevented MGO‐induced apoptosis in HAoECs by increasing anti‐apoptotic Bcl‐2 expression and decreasing pro‐apoptotic Bax expression. Pretreatment with LB strongly inhibited the activation of all MAPK signals. Our study suggests that LB prevents apoptosis induced by MGO in HAoECs via inhibition of the oxidative stress and MAPK signaling pathways. In conclusion, LB may prove beneficial for the treatment of diabetic complications.Support or Funding InformationThis research was supported by iPET (Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries), Ministry of Agriculture, Food and Rural Affairs (No.115045‐3)
Essential Opposite Roles of ERK and Akt Signaling in Cardiac Steroid‐Induced Increase in Heart Contractility
Authors: Lichtstein David , Rosen Haim , Ami Hagit Cohen‐Ben , Inbal Adi , Buzaglo Nahum 
Published: November 2023

Interaction of cardiac steroids (CS) with the Na+, K+‐ATPase elicits, in addition to inhibition of the enzyme's activity, the activation of intracellular signaling such as ERK and Akt. We hypothesized that the activities of these pathways are involved in CS‐induced increase in heart contractility. This hypothesis was tested using in‐vivo and ex‐vivo wt and SERCA1a deficient zebrafish (accordion, acc mutant) experimental model. Heart contractility was measured in‐vivo, and in primary cardiomyocytes, in wt zebrafish larvae and acc mutant. Ca2+ transients were determined ex‐vivo, in and adult zebrafish hearts. CS dose‐dependently, augmented the force of contraction of larvae heart muscle and cardiomyocytes and increased Ca2+ transients, in wt, but not in acc mutant. CS in‐vivo, increased the phosphorylation rate of ERK and Akt in the adult zebrafish heart of the two strains. Pretreatment of wt zebrafish larvae or cardiomyocytes with specific MAPK inhibitors completely abolished the CS‐induced increase in contractility. On the contrary, pretreatment with Akt inhibitor significantly enhanced the CS‐induced increase in heart contractility both, in‐vivo and ex‐vivo without affecting CS‐induced Ca2+ transients. Furthermore, pretreatment of the acc mutant larvae or cardiomyocytes with Akt inhibitor restored the CS‐induced increase in heart contractility also without affecting Ca2+ transients. These results support the notion that the activity of MAPK pathway is obligatory for CS‐induced increases in heart muscle contractility. Akt activity, on the other hand, plays a negative role, via Ca2+ independent mechanisms, in CS action. These findings point to novel potential pharmacological intervention to increase CS efficacy.Support or Funding InformationThis work was supported by grants from the Ministry of Trade and Industry (NOFAR), Israel and the Szolts Foundation, The Hebrew University of Jerusalem, and the Walter and Greta Chair in Heart Studies (to D.L).
Suvorexant, a Clinically Used Dual Orexin Receptor Antagonist, Induces Human Pregnane X Receptor‐Mediated Expression of Cytochrome P450 3A4
Authors: Abbott Kodye , Pondugula Satyanarayana 
Published: November 2023

Activation of human pregnane X receptor (hPXR)‐regulated expression of cytochrome p450 3A4 (CYP3A4) plays an important role in mediating adverse drug interactions. Here, we studied whether suvorexant, a dual orexin receptor antagonist used to treat insomnia, could induce hPXR‐mediated regulation of CYP3A4. hPXR transactivation assays were performed to determine CYP3A4 promoter activity and in HepG2 human liver carcinoma cells as well as in LS174T and LS180 human colon cancer cells. Quantitative RT‐PCR assays were conducted to study gene expression of CYP3A4 in human primary hepatocytes and hepatocells. Suvorexant, at its therapeutically‐relevant concentrations, induced hPXR transactivation of CYP3A4 promoter activity. Similar to a known PXR agonist rifampicin, suvorexant also induced CYP3A4 gene expression in human primary hepatocytes and hepatocells. These results suggest that suvorexant induces hPXR‐regulated CYP3A4 gene expression. As insomnia often coexists with other medical and psychiatric conditions, it is treated concurrently with the coexisting conditions. Therefore, the inductive effects of suvorexant on CYP3A4 expression caution the use of suvorexant in conjunction with other medications metabolized via CYP3A4. Future studies will be directed to investigate the mechanisms of suvorexant induction of hPXR‐mediated CYP3A4 expression.Support or Funding InformationThis work was supported by the Auburn University Research Initiative in Cancer Grant, Animal Health and Disease Research Grant, and Auburn University Startup Funds to Pondugula SR. We thank Drs. Tao, Coleman, and Schwartz for sharing their facilities.
Rutin ameliorates high cholesterol diet induced alteration in lipid profile Interleukin‐3 (IL3) and Interleukin6 (IL6) and liver inflammation
Authors: Alhosaini Khaled , AlSharari Shakir , Hafez Mohammed M. 
Published: November 2023

Non alcoholic fatty liver disease (NAFLD) has become a very common metabolic disorder. It refers to a group of conditions where excess fats are deposited in hepatic cells. High cholesterol diet (HCD) causes dyslipidemic syndrome and hyperlipidemia. Furthermore, hypercholesterolemia induceshepatic oxidative stress and inflammation and activation of inflammatory mediators and suppression of the anti‐inflammatory genes. The natural flavonoid (Rutin) possesses an antioxidant activity with protective potential. Therefore, we designed this study with the aim to investigate the potential ameliorative potential of rutin against HCD‐induced alteration of lipid profile and hepatic interleukins; Interleukin‐3 (IL3) and Interleukin6 (IL6). Male Wistar rats were allocated into four groups as follow (1) control (Rat chow), (2) Rutin (0.2% in rat chow), (3) HCD (1% cholesterol and 0.5% cholic acid in rat chow) and (4) rutin (0.2%) + HCD. Animals received freshly prepared experimental diets for 6 consecutive weeks. In plasma, levels triglyceride (TG), total cholesterol (TC), low‐density lipoprotein (LDL) and high‐density lipoprotein (HDL)were estimated. In hepatic tissue, gene expressions of Interleukin‐3 (IL3) and Interleukin6 (IL6) were determined using real time PCR. HCD significantly increased plasma levels of TG, TC and LDL by 48%, 89% and 67% respectively and significantly decreased the HDL levels by 17% compared to control group. Rutin supplementation in combination with HCD, significantly decreased TG and LDL levels and insignificant decrease plasma levels of TC and HDL compared to control group. Results also showed a significant increase by 5.8 fold in the expression of IL‐6 and significant decrease by 3 folds in IL‐3 expression in rats fed with HCD compared to control group. Interestingly, administration of rutin in combination with HCD corrected the alteration in the expression of these genes to their values in the control group. In conclusion: Combination of the natural occurring antioxidant, Rutin, with HCD protects against HCD associated hepatic inflammation and the altered lipid profile.Support or Funding Informationking saud university
TRPM8 Mediates the Anti‐inflammatory Effects of Eucalyptol in a Mouse Model of LPS‐Induced Pulmonary Inflammation
Authors: Caceres Ana Isabel , Liu Boyi , Jabba Sairam V. , Morris John B. , Jordt Sven‐Eric 
Published: November 2023

Eucalyptol (1,8‐cineol), the major ingredient in the essential oil of the eucalyptus leaves and other medicinal plants, has long been known for its anti‐inflammatory properties. Eucalyptol interacts with Transient Receptor Potential (TRP) ion channels among other targets, but it is unclear which of these mediates its anti‐inflammatory effects. In this work, the effects of eucalyptol were compared in WT and TRPM8‐deficient mice in a model of pulmonary inflammation following intranasal administration of lipopolysaccharide (LPS). LPS, a component of Gram‐negative bacterial cell walls, plays a major role in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). 24 hours after LPS intranasal instillation lungs and bronchoalvelolar lavage (BAL) were collected and analyzed for leukocyte infiltration, enzyme activities and cytokine/chemokine levels by qPCR, Luminex and cell counting among other techniques.Our results show that eucalyptol treatment diminished leukocyte infiltration, myeloperoxidase activity and production of TNF‐α, IL‐1β, IFN‐γ and IL‐6. Genetic deletion of TRPM8 abolishes the anti‐inflammatory effects of eucalyptol in the LPS model. Eucalyptol is at least 30‐fold more potent on human TRPM8 than on mouse TRPM8, and a metabolite of eucalyptol, 2‐hydroxy‐1,8‐cineol, also activates human TRPM8. This shows that among the pharmacological targets of eucalyptol, TRPM8 is essential for its anti‐inflammatory effects in mice, likely mediated by peripheral sensory neurons. The much higher sensitivity of human TRPM8 to eucalyptol compared to the rodent orthologues explains the higher potency of eucalyptol in humans. Metabolites of eucalyptol likely contribute to the anti‐inflammatory effects. The development of more potent and selective TRPM8 agonists may yield novel anti‐inflammatory therapeutics.Support or Funding Information‐ NIH grants R01HL105635 and R01HL105635‐S1 (to S. E. J. and J. B. M.)‐ P50DA036151 project #1 (to S.E.J.).
PHOSPHODIESTERASE (PDE) INHIBITORS AND THEIR INTERACTION WITH NITRIC OXIDE (NO) MODULATORS IN ADJUVANT‐INDUCED RHEUMATOID ARTHRITIS
Authors: Pal Rishi , Tiwari Prafulla Chandra , Nath Rajendra , Babu Suresh , Pant Kamlesh Kumar 
Published: November 2023

Phosphodiesterase (PDE) inhibitors are known to increase cAMP/cGMP in the cellular compartment and leads to signal transduction process in the various cell system. Here we focused on the anti‐inflammatory & immunomodulatory protective role of some PDE inhibitors theophylline, pentoxifylline & rolipram and their interactions with nitric oxide (NO) modulators in complete Freund's adjuvant induced rheumatoid arthritis in rats. Wistar rats (200–300 gm, n=6/group) of either sex were used in the study. On day ‘0’ rats were injected with 0.2 ml of complete Freund's adjuvant (CFA) in sub‐planter region of right hind paw along with 0.1 ml of squalene to develop RA while controls received only vehicle. PDE inhibitors drug treatment alone and in combination with NO modulators was given from day ‘14’ to ‘28’. Arthritic parameters a) arthritis index, b) ankle diameter c) paw volume and their body weight were noted to evaluate progression of RA on day 0, 7, 14, 21 and 28. On day ‘28’ rats were sacrificed and their blood and paws were collected for TNF‐α and IL‐10 cytokine estimation, pathological examination and NF‐kB expression. Data obtained was analysed using two‐way ANOVA followed by Newman‐Keul's posthoc test and p<0.05 was considered for significance. It was found that CFA significantly increased arthritis‐index, paw volume, ankle diameter and serum TNF‐α and NF‐kB levels while body weight and serum IL‐10 levels was significantly decreased (P<0.05). These CFA‐induced changes were significantly reversed by theophylline (10 & 20 mg/kg), P<0.0001); pentoxifylline (5 & 10 mg/kg), P<0.002); rolipram (1 & 2 mg/kg), P<0.05) alone and in combination with L‐arginine (100 mg/kg) and or L‐NAME (10 mg/kg) in dose dependent manner (p<0.005) in all parameters). The maximum protective effects was observed in theophylline > pentoxifylline > rolipram with L‐NAME 10 mg/kg combination treatment group (p<0.001). The data obtained was substantiated histopathologically. Results of this study are suggestive of the involvement & interaction of NO with different PDE inhibitors may have protective role in adjuvant induced RA with may be mediated through cytokine signalling pathways.Support or Funding InformationThe research work financially supported by research cell, King George's Medical University, UP, Lucknow in the form of Intramural research grant awarded to First author Dr. Rishi Pal.
Neutrophil‐derived 20‐HETE Is a Major Determinant of Coronary Collateral Growth
Authors: Joseph Gregory , Hutcheson Brenda , Hunter Ian , Soler Amanda , Gotlinger Katie , Falck John , Schwartzman Michal L , Rocic Petra 
Published: November 2023

Coronary collateral growth (CCG) is an adaptive response to transient, repetitive coronary artery occlusion in normal animals but is markedly impaired in metabolic syndrome. Recent studies in our laboratory have shown that in metabolic syndrome rats (JCR:LA‐cp, JCR) in addition to endothelial cell (EC) dysfunction, neutrophil survival is enhanced, leading to neutrophil accumulation which contributes to compromised CCG in JCR rats. 20‐hydroxyeicosatetraenoic acid (20‐HETE) plays an important role in vascular function. However, the role of 20‐HETE in the regulation of CCG is unknown. We investigated the hypothesis that elevated neutrophil‐derived 20‐HETE promotes EC dysfunction and apoptosis and impairs CCG in metabolic syndrome. In our study, coronary blood flow in Sprague‐Dawley (SD) or JCR rats was measured in the collateral (CZ) and normal zone (NZ) of the heart after 9 days of transient, repetitive LAD occlusion (repetitive ischemia, RI) using microspheres. Collateral growth in the JCR rats was impaired compared to SD rats (CZ/NZ flow ratio was 0.11±0.02 in JCR vs. 0.84±0.02 in SD). Administration of 20‐HETE antagonists, either 20‐SOLA or 20‐HEDGE, fully restored collateral‐dependent blood flow (0.85±0.01 in JCR+20‐SOLA, 0.84±0.01 in JCR+20‐HEDGE), while neutrophil depletion, via neutrophil‐blocking antibodies, partially restored collateral‐dependent blood flow (0.57±0.02) in JCR rats. Neutrophil accumulation was ~3 times higher and persisted for the duration of the RI protocol in the CZ of JCR vs. SD rats, where their infiltration was low and transient. This RI‐induced, elevated neutrophil infiltration in the CZ of JCR rats correlated with a >4‐fold increase in CYP4F (the neutrophil 20‐HETE CYP isoform) expression and a 4‐fold increase in 20‐HETE levels vs. SD rats, which was abolished by administration of neutrophil‐blocking antibodies to JCR rats. Furthermore, coronary microvessels of JCR rats displayed reduced endothelium‐dependent vasodilation after 9 days of RI (20%±1.2% JCR vs. 40%±2.3% SD), which was reversed with neutrophil‐blocking antibodies (37%±2.5% JCR+anti‐CD11b/CD18/CD44). eNOS phosphorylation (p1179) and NO production were likewise decreased (~50%) in the CZ of JCR rays after 9 days of RI, which was reversed after administration of neutrophil‐blocking antibodies. Lastly, RI‐induced EC apoptosis in JCR but not in SD rats (TUNEL). EC viability in JCR rats was almost completely restored by treatment with neutrophil‐blocking antibodies. Taken together, these results indicate that neutrophil‐derived 20‐HETE is a major contributor to impaired EC viability and function, and thus to impaired CCG in metabolic syndrome.Support or Funding InformationSupport: NIH R01 HL093052.
How Important Is SERCA In Neuronal Calcium Homeostasis?
Authors: Shanks Julia , Paterson David 
Published: November 2023

Maintaining intracellular calcium homeostasis is critical for excitation‐contraction coupling in the heart, and is essentially dependent on primary and secondary active transport systems. However, upregulating sarcoendoplasmic reticulum calcium ATPase (SERCA) in cardiomyocytes from patients with heart failure has recently proved inconclusive (CUPID2 trial). Neuronal calcium impairment is also seen in cardiovascular diseases resulting in heightened sympathetic responsiveness that further exacerbate the disease phenotype. We tested whether overexpression of SERCA in cardiac sympathetic neurons could modulate intracellular calcium transients. Isolated neurons were incubated with either Ad‐MCherry‐SERCA2A or Ad‐MCherry‐Empty (control) for 24 hours immediately post isolation. Calcium imaging using fura‐2AM (2.5μM) was carried out three days post transfection. SERCA2A transgene neurons showed greater depolarisation induced intracellular calcium transients, compared to control (Δ 380/340 ratio (au): SERCA 0.75±0.03, n=68; Empty 0.64±0.03, n=57. P<0.01). SERCA2A expressing neurons also showed greater SR calcium storage compared to control, as demonstrated by significantly higher calcium responses to caffeine (10mM: Δ 380/340 ratio (au): SERCA 0.15±0.01, n=45; Empty 0.03±0.01, n=35. P<0.0001) and thapsigargin stimulation (1μM: Δ 380/340 ratio (au): SERCA 0.12±0.02, n=42; Empty 0.03±0.01, n=33. P<0.0001). Additionally mitochondrial calcium uncoupling with FCCP (1μM) resulted in a greater intracellular calcium change in the SERCA2A transgenic neurons (Δ 380/340 ratio (au): SERCA 0.13±0.01, n=22; Empty 0.05±0.01, n=22. P<0.0001), indicating that altering neuronal calcium handling with upregulation of SERCA has also effected mitochondrial calcium handling and potentially metabolic function in these neurons. Within this study we demonstrated that a SERCA2A gene construct can be transfected in isolated sympathetic neurons from the stellate ganglia. Moreover, this transduction caused an increase in intracellular calcium transients that would be expected to facilitate neurotransmission and be potentially deleterious for cardiac function.Support or Funding InformationBritish Heart Foundation
Combining Mass Spectrometry Cross‐Linking, Molecular Docking and NMR Titration to Dissect ZapA‐FtsZ Protein Interaction
Authors: Nogueira Maria Luiza Caldas , Sforça Mauricio Luis , Leme Adriana Franco Paes , Pauletti Bianca Alves , de Oliveira Paulo Sergio Lopes , Honorato Rodrigo Vargas , King Glenn F , Filho Frederico José Gueiros , de Mattos Zeri Ana Carolina 
Published: November 2023

The cell division is a key event in a bacteria cycle of life and in most rod shaped bacteria. This cell divion involves a septum formation. FtsZ and other proteins forms a macromolecular complex, named Divisome, which redirect the cell wall and provide a constrictive force required to divide the bacterial cell in two daughter cells. FtsZ is a tubulin homologue that polymerizes as a ring structure shaped, the Z ring. The Divisome is composed by: the Z ring, modulators proteins, proteins that anchor the Z ring to the cell membrane and synthesize cell wall (peptidoglycan). Modulators proteins are essential for the cell division occurs at the right place and time, because they act stimulating or preventing the Z ring formation. Defects on modular proteins could cause defects in bacterial cell division such as filamentation and mini‐cell formation. The Z ring associated protein A (ZapA) is a positive modulator that assists the bundling of FtsZ polymers by promote lateral association of FtsZ filaments. Despite the interaction between FtsZ‐ZapA was described many years ago, the molecular mechanisms details involved into this process is barely understood. Here we combined the information obtained from FtsZ‐ZapA NMR titration and Mass Spectrometry cross‐linking to modelling the FtsZ and ZapA protein complex. The docking model may give us a clue of the main amino acids residues involved on that interaction and how thus it occurs.Support or Funding InformationSão Paulo Research Foundation (FAPESP); State University of Campinas (UNICAMP) and Brazilian Center for Research in Energy and Materials/Brazilian Biosciences National Laboratory ( CNPEM/LNBio )