bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Piezo1 in cAMP‐Dependent Calcium Release From ER Stores in Endothelial Cells
Authors: Santana Dianicha , Malik Asrar B. , Mehta Dolly , Komarova Yulia A. 
Published: June 2021

Endothelial cells (ECs) lining the vessel wall express various mechanosensing and mechanotransducing molecules that are responsible for converting mechanical cues into chemical signals to elicit specific cellular responses. The recently discovered mechanosensitive Ca2+ channel Piezo1 plays an important role in sensing hydrostatic pressure and shear stress by ECs. However, specific mechanisms involved in Piezo1‐mediated mechanotransduction remains elusive. Using intracellular and endoplasmic reticulum (ER) calcium sensors to respectively monitor the changes in Ca2+I and Ca2+ ER concentrations, we have shown that pharmacological activation of Piezo1 induces cAMP‐evoked Ca2+ release from ER stores. Ca2+influx through Piezo1 leads to a transient activation of soluble adenylyl cyclase (sAC) which, in turn, activates cAMP‐dependent Ca2+ release via Inositol TrisphosphateReceptor 2 (IP3R2), a calcium channel located in ER. Depletion of either sAC or IP3R2 markedly reduces the Piezo1‐mediated increase in cytosolic Ca2+ I indicating that Ca2+ ER release represents a positive feedback loop amplifying Ca2+ signals in ECs. Our data, for the first time, demonstrate that mechanosensitive channels induces Ca2+ release from the ER. Further work will establish the role of sAC and IP3R2 in regulating the adaptive cellular responses of ECs to mechanical cues.Support or Funding InformationThis work is supported by the National Institutes of Health T32HL007829 to DS and 2R01HL045638‐30 to ABM and YK.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Caffeic Acid Phenethyl Ester Improves Diabetic Mice Cutaneous Wound Healing
Authors: Salles Jeanine , de Souza Bruna Romana , Costa Andréa Monte Alto 
Published: June 2021

Diabetes mellitus is a chronic condition that causes several complications. Studies indicate that around 1/3 of the diabetic patients in USA suffer cutaneous complications, resulting in a higher morbidity and mortality. In the last years, scientific studies have used the active principle of the natural products to treat acute and chronic wounds. As an example we have the caffeic acid phenethyl ester (CAPE), one of the propolis active principles, that presents potent antioxidant and anti‐inflammatory action. Due the great performance of CAPE to improve the burn healing in rats, as observed in our laboratory, we seek to investigate if this agent can also be beneficial in wound healing of chronic lesions. Diabetes was induced in Swiss mice (males) by multiple streptozotocin injections. After one week, hyperglycemia was confirmed and two excisional wounds (8 mm2 each) were made on the dorsum of these animals (day 0). Mice were divided in two groups, the diabetic mice group (DB group), that received only the vehicle and the diabetic mice CAPE treated‐ group (DB+CAPE group), that received 10μM of CAPE dissolved in the vehicle. The treatment started on the day of the wounds creation and extended until the day of euthanasia, 12 days after wounding, when the wounds were collected. CAPE treatment improved wound inflammatory and oxidative profile; in this group the tumor necrosis factor α (TNF‐α) and phosphorylated nuclear factor kappa B (NFkBp65) protein, nitric oxide synthase 2 (NOS2) and cyclooxygenase‐2 (COX‐2) expression was reduced. These results reflected in the inflammatory cell infiltrate, which was reduced. The reduction in inflammatory cell amount in DC+CAPE group influenced the reduction of the protein carbonylation and lipid peroxidation. In DB+CAPE group, the granulation tissue presented a greater collagen fiber density, with more collagen type I. The improvement in inflammatory parameters influenced the macroscopic parameters. The DB+CAPE group presented a strong tendency towards greater contraction and reepithelization when compared to DB group. These results lead us to believe that the treatment of diabetic lesions with CAPE may be a good option for a therapeutic treatment, with the benefit of being a product of natural origin, which could reduce side effects.Support or Funding InformationFundação de Amparo à Pesquisa do Estado do Rio de Janeiro ‐ FAPERJThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Oral Administration of a Vermiculite‐Nanocellulose Hybrid Boosts Sperm Parameters and Gonadotropin Levels in Rats
Authors: Otuechere Chiagoziem A. , Adewuyi Adewale , Oluwabayo Tanitoluwa , Afolayan Folashade , Avwioroko Oghenetega , Abazuh Uche 
Published: June 2021

The safety of vermiculite‐nanocellulose (VEN) hybrid on the male reproductive system is not known. Hence we investigated the effects of VEN on the biochemical and hormonal indices of rats. Animals were orally exposed to VEN at 0, 5, 10 and 20 mg/kg body weight for 14 consecutive days. Administration of VEN had no effect on organo‐somatic index of the testes. Although Gamma –glutamyl transferase, catalase and superoxide dismutase activities and hydrogen peroxide level remained unaffected, exposure of rats to VEN significantly depleted myeloperoxidase, glutathione peroxidase and malondialdehyde levels in the testes of treated rats, when compared to control. The spermiogram of VEN‐exposed rats showed a significantly higher sperm count and lower sperm morphology rates than control group. Treatment with VEN also significantly increased circulatory concentrations of follicle‐stimulating hormone and luteinizing hormone, and decreased testosterone levels at the 20 mg/kg dose. Histoarchitecture of the testes, in the treatment groups, except at the 20 mg/kg dose, revealed normal looking Leydig and Sertoli cells. The epididymis presented with tubules completely filled with spermatids across all treatment groups. Taken together, our study show that VEN could play an important role in male reproduction in rats, especially in redox regulation and stimulation of the gonadotropins.Support or Funding InformationThis research was done without specific grant from any funding agency in the public, commercial, or not‐for‐profit sectors.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Cataractogenesis in Cultured Bovine Lenses by ATB 343
Authors: Maffofou Leonce N , SegewkalHeruye Segewkal H , Njie‐Mbye Ya Fatou , Ohia Sunny E , Opere Catherine A. 
Published: June 2021

PurposeThere is evidence that substrates for the endogenous production of hydrogen sulfide (H2S) such as L‐cysteine and N‐acetylcysteine (Zhang et al., Mol Vis, 14:862–870, 2008) and NSAIDs such as ibuprofen (Robert and Harding, Exp Eye Res, 54:509–518) can mitigate cataractogenesis in vitro and in vivo. However, the role of the hybrid compound consisting of the NSAID, indomethacin and a H2S‐donating moiety, ATB 343 on cataractogenesis has not been elucidated. In this study, we investigated the effect of the ATB 343 on cataractogenesis in cultured bovine lenses.MethodsFreshly isolated bovine lenses were cultured in a DMEM buffer solution as follows: Group I: Control (DMEM); Group II–III: H2O2 (10 mM or 50 mM); Group IV–VI: ATB 343 (10−8M to 10−6M; Group VII: ascorbic acid (AA; 10 mM) in presence and absence of H2O2 (10 mM or 50 mM). Lenses were incubated in a CO2 chamber and assessed at 3, 6, 24, 48 and 72 hour‐time points. Qualitative assessments were conducted by visual inspection of lenses and photographic images captured against a black grid, while quantitative assessment was conducted by measurement of transmittance using a plate reader (Synergy H1 hybrid reader; Bio Tek Instruments, Inc) at every time point.ResultsDMEM‐cultured lenses exhibited a time‐dependent decrease in transmittance (420nm) and a corresponding loss of lens optical clarity up to 72 hours. Similar to the endogenous antioxidant, AA (10 mM), ATB 343 (10−7M) attenuated time‐dependent lens degradation up to 24 hours. H2O2 (10 mM and 50 mM) decreased light transmittance in a time‐dependent manner, achieving an inhibition of 41.98% (n=12; p<0.0001) and 46.37% (n=12; p<0.0001), respectively after 72 hours. Interestingly, ATB 343 (10−8 M to 10−6 M) partially reversed H2O2 (10 mM)‐induced degradation up to 72 hours. After 48 hours, the rank order of protection was: ATB (10−7M)>ATB (10−6M)>AA (10 mM)>ATB (10−8M). Similarly, ATB 343 (10−7 M and 10−6M) partially reversed H2O2 (50 mM)‐induced lens degradation up to 48 hours. After 24 hours, the rank order of protection was: ATB (10−7M)>ATB (10−8 M)>ATB (10−6M >AA (10 mM).ConclusionATB 343 protected cultured lenses from time‐dependent degradation up to 24 hours and partially protected cultured bovine lenses from H2O2‐induced cataractogenesis.Support or Funding InformationCreighton University‐SPAHP, Pharmaceutical Sciences Graduate ProgramThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Nox4 NADPH Oxidase Splice Variants Generate Hydrogen Peroxide and Modify the Cell Cycle
Authors: Scott Danielle A , Schickling Brandon M , Miller Francis J 
Published: June 2021

The NADPH oxidase (Nox) family of proteins generate reactive oxygen species (ROS), which regulate redox‐mediated cell signaling and may lead to oxidative stress. Unlike the other catalytic subunits, Nox4 is mostly constitutively active and regulated by its expression levels. It is reported that Nox4 has splice isoforms, including variant D which retains all FADH and NADPH binding sites of the full length Nox4A, but lacks transmembrane domains. The goal of this study was to characterize expression and function of splice variants Nox4A and Nox4D. We first validated the specificity of qPCR primers used for detection of the splice variants by overexpression of Nox4A and Nox4D in Cos22 cells. Next, we examined expression of these isoforms in cancer cell lines. The relative expression of Nox4A and Nox4D was not significantly different in human epithelial colorectal adenocarcinoma cells (CaCo), human Asian endometrial adenocarcinoma (Ishikawa) glioblastoma (LN229), adenocarcinomic human alveolar basal epithelial (A549) and human umbilical vein endothelial (EA.hy926) cells. We next examined the function of Nox4 splice variants and found that expression of Nox4A and Nox4D both increased ROS levels as compared to control transfected cells. It has been suggested that the subcellular targeting of Nox4A and Nox4D may be different. To test this, we measured hydrogen peroxide levels in the cytosol, mitochondria, and nucleus using the genetically targeted HyPer probes. Whereas Nox4A expression increased ROS in each subcellular compartment of Cos22 cells, Nox4D primarily increased the HyPer signal in the nucleus. We next examined the effect of Nox4 on protein thiol oxidation and found that expression of Nox4A caused oxidation of several protein thiols. Finally, Cos22 cells were transfected with Nox4A, Nox4D, or an empty vector and treated with cytokines. The expression of Nox4 isoforms had a differential effect on the percentage of cells in the G0/G1, S, and G2/M cell cycle phase. In summary, Nox4A and Nox4D splice isoforms are detectable in different cell lines, overexpression of Nox4A and Nox4D increase ROS levels in subcellular organelles and the oxidation of multiple protein thiols, and expression of Nox4A and Nox4D alters the cell cycle. The expression of Nox4 splice isoforms may provide specificity for regulation of subcellular redox signaling.Support or Funding InformationDAS was supported by the American Society for Pharmacology and Experimental Therapeutics (ASPET) sponsored Duke SURPH program. FJM is supported by the Office of Research and Development, Department of Veterans Affairs [2I01BX001729] and the National Institutes of Health [HL130039].This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sorafenib induces autophagic cell death and apoptosis in hepatoma HepG2 cells via PERK/EIF2α signaling pathway
Authors: Liu Ze‐feng , Liu Qi‐Lin , Li Fu‐Jian , Yang Zhou‐Fan , Chen Li , Zhang Guo 
Published: June 2021

Backgound & AimsPrevious studies have documented cellular apoptosis, autophagy and endoplasmic reticulum (ER) stress participated in the hepatocellular carcinoma (HCC). Sorafenib has been reported to regulate autophagy and apoptosis in hepatoma cells associated with different unfolded protein response (UPR). However, resistance problem dramatically limited the usage of sorafenib in HCC with uncertain mechanism. Therefore, this research is focused on whether and how sorafenib induces autophagy and apoptosis via specific PERK/EIF2a signaling pathway of ER stress in HepG2 cells.MethodsFirstly, stable infected HepG2 cell lines of PERK knockdown and vehicle control cells were established with lentivirus‐mediated RNA interference, RT‐PCR and western blot were employed to identify the efficiency of PERK silencing. Trypen blue assay was then applied to investigate the effect of PERK knockdown, and sorafenib (10 μM) induced‐lethality in these established HepG2 cell lines in presence or absence of thapsigargin (Tg, 500 nM). Western blot was further performed to analyze the protein expression of PERK, p‐PERK, EIF2α, p‐EIF2α, CHOP, caspase3 and P62. Finally, the numbers of apoptotic cells in these cells were measured by flow cytometry.ResultsAfter PERK silencing, the expression of PERK mRNA and protein were significantly lower (P<0.05) in the established HepG2 cell lines v.s. control. In parallel, the expression of p‐PERK, EIF2α, p‐EIF2α, CHOP and cleaved caspase3 protein were significantly decreased (P<0.05) and apoptotic numbers were dramatically inhibited in PERK silencing cells (P<0.05). In basal condition, sorafenib was not only found to induce autophagic flux but also drive up‐regulation of PERK, p‐PERK, EIF2α, p‐EIF2α, CHOP in vehicle HepG2 cells. Interestingly, PERK overexpression further induced autophagy with decrement in P62 in HepG2 cells in response to sorafenib or sorafenib plus Tg (P<0.05). Meanwhile, trypan blue assay found that the sorafenib alone and sorafenib combined with Tg induced‐cell death was substantially decreased with about 10% and 11% in PERK silencing cells but increased with 30% in PERK overexpression cells, respectively (P<0.05).ConclusionPERK, one of the hallmarks of ER stress signaling, might cause procelldeath autophagy and thereby contribute to enhance sorafenib derived apoptosis in hepatoma HepG2 cells, which may provide a potential critical target for overcoming the resistance of sorafenib in HCC in the future.Support or Funding InformationSupported by NSFC Grants 81360077, 81860654‐30630145, 81172260 and GXNSFC Grants 2015GXNSFDA139022 to Guo ZhangThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Antioxidant and Cytotoxic Properties of Tridax Procumbens Leaf Extract on Liver and Pancreatic Cancer Cell Lines
Authors: Samuel Ekundayo Stephen , Akinola Tolulope Joseph , Tosh David , Odunola Oyeronke Adunni 
Published: June 2021

Human hepatocellular carcinoma and pancreatic ductal adenocarcinoma are two of the deadliest emerging forms of cancer worldwide. Hence, the increased search for phytochemicals with promising anticancer properties. In the present study, we assessed the phytochemistry, antioxidant activities and the role of Tridax procumbens leaf extract and fractions on HepG2 and Panc‐1 cells (models of hepatocellular and pancreatic ductal adenocarcinoma). HepG2 and Panc‐1 cell lines were maintained in supplemented Dulbecco's modified eagle's medium at 37°C in a humidified incubator with 5% CO2. The HepG2 and Panc‐1 cells were treated with 20, 50, 100 and 250 μg/ml of dimethyl sulfoxide (control), extract and fractions (n‐hexane, dichloromethane, ethyl acetate, aqueous) for 24 and 48hours. Phytochemistry (total flavonoid, total phenol) and antioxidant (2,2‐diphenyl‐1‐picrylhydrazyl hydrate, ferric reducing antioxidant power, nitric oxide radical, total antioxidant capacity) assays were performed on the test samples. The expression levels of catalase, alkaline phosphatase, glutathione S transferase‐pi and caspase 3 were monitored with immunofluorescence. Live/Dead and MTT assay were used to determine cytotoxicity of cultured cells. The dichloromethane and ethyl acetate fractions showed significant higher antioxidant activities as compared to other test samples. The dichloromethane fraction caused a significant level of cytotoxicity and induction in the activity of ALP, catalase and caspase 3 as compared to the control. Conversely, there was a significant cellular reduction and nuclear translocation of GST‐pi in dichloromethane fraction treated cell as compared to the control. The extract and fractions of Tridax procumbens have antioxidant activities. However, the dichloromethane fraction's activities depict a promising anticancer agent.Support or Funding InformationCommonwealth Scholarship Commission, Department for International Development, United Kingdom and University of Ibadan, Ibadan, NigeriaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Study the Effects of Capsaicin on Triple Negative Breast Cancer Cells
Authors: Alasmari Moudi M. , Alshaeri Heba K. , Alashari Rawabi A. , Böhlke Mark , Maher Timothy , Pino‐Figueroa Alejandro 
Published: June 2021

Triple negative breast cancer (TNBC) is one of the most aggressive types of breast cancer accounting for 12% of breast cancer cases. It is characterized by the lack of the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER‐2), which limits treatment options and enhances its ability to metastasize with the risk of recurrence. Patients with TNBC are not responsive to conventional therapies. Capsaicin (CAP) is the most abundant capsaicinoid produced in chili pepper fruits and has been utilized for its analgesic and anti‐inflammatory effects. While several studies have demonstrated that capsaicin has anti‐carcinogenic properties in various types of human cancers, the underlying molecular mechanisms remain to be explored. The aim of this study was to investigate the effects of capsaicin in human TNBC, utilizing a BT‐20 cell model. Capsaicin demonstrated concentration‐ and time‐dependent in the viability of BT‐20 cells, as determined by the MTS assay. Capsaicin caused significant increases in cytochrome C release, caspase 3/7 activity and expression of cleaved poly‐(ADP‐ribose) polymerase (PARP), all of which are markers of apoptotic activation. These effects were accompanied by down‐regulation of cyclin D1 production, an indicator of cell cycle arrest at the G0/G1 phase. Further analyses of signaling mechanisms revealed that capsaicin significantly inhibited EGFR phosphorylation and the phosphorylation of its downstream signaling proteins AKT and MAPK, which may provide some explanation for its mechanism of action in TNBC. In conclusion, capsaicin has demonstrated inhibitory effects on cell growth and cell cycle progression on TNBC cells by enhancing apoptosis. Our data demonstrated a novel mechanism for capsaicin, modulating EGFR signaling by AKT/MAPK pathways in BT‐20 cells. These results provide useful information relevant to the development of a potential new therapy to treat TNBC with capsaicin.Support or Funding InformationThis work was funded by the College of Medicine, King Saud bin Abdulaziz University for Health Sciences (KSAU‐HS), Jeddah, Saudi Arabia.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Prevention of Cardiac Dysfunction Development by a Methanocarba Adenosine Monophosphonate Derivative
Authors: Shen Jian‐Bing , Toti Kiran S , Chakraborty Saibal , Kumar T. Santhosh , Cronin Chunxia , Liang Bruce T , Jacobson Kenneth A 
Published: June 2021

Accumulating evidence supports a therapeutic role of purinergic signaling in cardiac diseases. Previous studies showed that MRS2339, a charged methanocarba derivative of 2‐Cl‐adenosine monophosphate, is efficacious in animal models of heart failure when infused systemically. However, MRS2339 lacks oral bioavailability as a nucleoside 5’‐phosphate derivative and is subject to hydrolysis in vivo leading to the inactive nucleoside MRS1873. We now test the hypothesis that uncharged adenine nucleoside phosphonates, suitable as oral agents with a hydrolysis‐resistant phospho moiety, can prevent the development of cardiac dysfunction in a post‐infarction ischemic or pressure overloaded heart failure model. Of the diester‐masked uncharged phosphonates evaluated, MRS2978 but not MRS4074, was efficacious in preventing cardiac dysfunction with improved left ventricular (LV) fractional shortening (FS increased from 21±0.79% with vehicle (N=13) to 25.1±0.63% with MRS2978 (N=11), P<0.05) and with preservation of LV wall thickness when administered orally at onset of ischemic heart failure (within 24h after left anterior descending artery (LAD) ligation). MRS2925, the charged, unmasked analog of MRS2978, was able to prevent heart dysfunction only when it was infused subcutaneously; its oral gavage was not efficacious. In the established heart failure models caused either by LAD ligation or transverse aortic constriction (TAC), orally administrated MRS2978 (4 weeks after LAD ligation or 3 weeks after TAC), but not MRS2925, could partially restore cardiac dysfunction by increasing the FS in both ischemic (from 19.7±0.95% with vehicle (N=18) to 26.1±1.37% with MRS2978 (N=11), P<0.05) and pressure overload (FS from 17.5±0.76% with vehicle(N=9) to 23.7±1.25% with MRS2978 (N=10), P<0.05) heart failure models. MRS2978 was orally bioavailable (57–75%F) and highly stable in human, mouse and rat plasma, with 96–100% remaining after 2 h. MRS2925 could be formed from hydrolysis of MRS2978 in vivo, but as a minor metabolite (only 10%), suggesting that the effects of MRS2978 are not dependent on the MRS2925. Either MRS2978 or MRS2925, combining with 2me‐SATP, was capable of activating cardiac myocyte eNOS, consistent with myocyte eNOS activation as a mechanism for the salutary effect of these two phosphonates nucleotides. These data demonstrated the ability of oral therapy with MRS2978 to treat heart failure, both at the inception of heart failure for prevention and after the development of heart failure as a rescue.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.