bluuebunny/crossref_metadata_2025_split huggingface dataset:
Angiontensinergic Receptors in the Medial Amygdaloid Nucleus Is Involved in Anxiogenic‐Like Effect Evoked by Emotional Stress in Rats
Authors: Crestani Carlos Cesar , Costa‐Ferreira Willian , Gomes‐de‐Souza Lucas 
Published: June 2021

The medial amygdaloid nucleus (MeA) is part of limbic circuit implicated in responses to stress. Besides, angiotensinergic neurotransmission has been described as a prominent neurochemical mechanism involved in stress‐evoked physiological and behavioral responses. However, the involvement of angiotensinergic neurotransmission within the MeA in behavioral responses evoked by exposure to aversive threats has never been investigated. Therefore, the aim of this study was to investigate the involvement of angiotensinergic neurotransmission within the MeA in anxiogenic‐like effect evoked by an acute session of restraint stress in rats. For this, male Wistar rats (240g–260g) had cannula‐guide bilaterally implanted into the MeA. The restraint stress was performed by placing the animals in a plastic cylindrical tube for 60 minutes. Independent group of animals received bilateral microinjections into the MeA of angiotensin II (0.05nmol/100nL), the selective AT1receptor antagonist losartan (1nmol/100nL), the selective AT2receptor antagonist PD123319 (0.05nmol/100nL), angiotensin 1–7 (0.05nmol/100nL), the selective Mas receptor antagonist A‐779 (0.1nmol/100nL) or vehicle (saline, 100nl)10 min before the onset of the restraint stress session. Immediately after the end of the restraint session, the anxiety‐like behavior was assessed in the elevated plus maze (EPM). Animals of naïve group (non‐stressed) were tested in the EPM together with stressed animals. We identified that restraint stress decreased the percentage of time spent (F(6,63)=7.98; P<0.0001) and number of entries (F(6,63)=7.40; P<0.0001) in the open arms of the EPM in the vehicle‐treated animals, when compared with naïve animals. Restraint did not affect the number of entries in the enclosed arms (F(6,63)=0.30; P>0.05) and the time spent in the center (F(6,63)=0.85; P>0.05). MeA treatment with either losartan, angiotensin II or angiotensin 1–7 inhibited the restraint‐evoked decrease in the percentage of time spent (P<0.05) and number of entries (P<0.05) in the open arms. Microinjection of either PD123319 (time: P<0.001; entries: P<0.0007) or A‐779 (time: P<0.008; entries: P<0.01) did not affect the decrease in the exploration of the EPM open arms evoked by restraint stress. In conclusion, our results indicate that AT1receptors within the MeA is involved in anxiogenic‐like effect of emotional stress. Besides, present data suggest that activation of the Mas receptor within the MeA inhibits this behavioral response.Support or Funding InformationFAPESP, CNPq and PADC‐FCF/UNESPThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Design, Synthesis, In Vitro, and In Silico Evaluation of a Novel Series of Serotonin 5‐HT2C Receptor (5‐HT2CR) Positive Allosteric Modulators (PAMs)
Authors: Wold Eric A. , Wild Christopher T. , Chen Jianping , Pazdrak Konrad , Garcia Erik J. , Anastasio Noelle C. , White Mark A. , Chen Haiying , Cunningham Kathryn A. , Zhou Jia 
Published: June 2021

Signal transduction and the modulation thereof characterize G protein‐coupled receptor (GPCR) function in the central nervous system (CNS) while CNS GPCRs are the therapeutic targets for most neuropsychiatric disorders. Endogenous ligands (e.g., neurotransmitters) and GPCRs are embedded into a dynamic landscape of temporally and spatially controlled signalling events that contribute to physiological brain function, and dysregulation of GPCR systems is implicated in pathophysiological states. Utilizing studies of the serotonin (5‐HT) 5‐HT2C receptor (5‐HT2CR), which has been implicated in CNS disorders such as drug use disorders and depression, we sought to discover novel modulators of 5‐HT2CR function that maintain the temporal and spatial characteristics of CNS signaling. Decreased 5‐HT2CR signaling has been implicated in dysregulated impulse control and drug‐seeking in rodent preclinical models, with selective 5‐HT2CR agonists exhibiting affinity to dampen these behavioral outcomes. The utilization of positive allosteric modulators (PAMs) is an attractive strategy to increase 5‐HT2CR functional response while limiting disruption of its dynamic signaling landscape. Our PAM discovery efforts have focused on two unique molecular scaffolds (i.e., CYD‐1‐79 and oleamide) and the derivatization of each. Synthesized, novel small molecules were screened via a cell‐based, 5‐HT‐induced, intracellular calcium (Ca2+) release assay with physiologically relevant 5‐HT2CR expression levels. Our studies have resulted in the discovery of multiple 5‐HT2CR PAMs, including CYD‐1‐79 and oleamide‐based molecular scaffolds, that increase 5‐HT2CR‐dependent function by 20% or greater. In vitro distribution, metabolism, and pharmacokinetic (DMPK) analyses and physicochemical properties indicate that our 5‐HT2CR PAMs display a range of promising lead‐ and drug‐like attributes. In silico molecular docking with the recently published 5‐HT2CR X‐ray crystal structure has identified a potential PAM binding site common to a diverse collection of small molecule 5‐HT2CR PAM derivatives. In conclusion, we have discovered and further optimized novel series of 5‐HT2CR PAMs with the potential for translation towards neurotherapeutics for drug use disorders and other CNS disorders in which 5‐HT2CR dysfunction is implicated as a mediator.Support or Funding InformationSupported by grants R01 DA038446 (JZ/KAC), K05 DA020087 (KAC), P30 DA028821 (KAC), T32 DA07287 (CTW, EAW), F31 DA038922 (CTW), F31 DA045511 (EAW)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Influence of Beta‐2 Adrenergic Receptor Genotype on Age of Nocturnal Ventilation in Patients with Duchenne Muscular Dystrophy
Authors: Kelley Eli , Cross Troy , Snyder Eric 
Published: June 2021

Duchenne muscular dystrophy (DMD) is an inheritable disease that presents itself in diminished respiratory function and is the main contributor to mortality in these patients. The main respiratory concerns in DMD are reduced diaphragmatic function, reduced mucocilliary clearance, increased muscle proteolysis, and increased inflammation. The Gly16 variant of the β2‐adrenergic receptor (ADRB2) has been identified as providing a protective effect for respiratory weakness. The aim of this study was to identify the influence of ADRB2 genotype on the age of use for nocturnal ventilatory assistance in DMD. Data from CINRG‐DNHS including 175 DMD patients (ages 3–25 years) with up to 9.7 years follow‐up were analyzed focusing on ADRB2 functional variants. We performed Cox proportional hazard and Kaplan‐Meier time to event analyses for the age of nocturnal ventilation use. There were no differences between genotype groups in age, height, weight, corticosteroid use, number of ambulatory patients, or age of loss of ambulation. The genotype groups differed significantly in the age at which nocturnal ventilation use was prescribed with the Gly16 demonstrating a 2.77‐fold increased risk of using nocturnal ventilation at any given age. These data suggest that genetic variations in the ADRB2 gene may influence the age of nocturnal ventilation use in DMD. Specifically, the Gly16 polymorphism of the ADRB2 may have a deleterious effect on age of nocturnal ventilation use in DMD.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel Compound Inhibits Pancreatic Cancer Cell Invasion, Tumor Sphere Formation, and In Vivo Tumor Growth in Mice by Suppressing EMT
Authors: Wang Tao , Dong Ruochen , Chen Ping , Baltezor Michael , Weir Scott , Chen Qi 
Published: June 2021

ObjectiveEpithelial to Mesenchymal Transition (EMT) has been proposed to contribute importantly to metastasis, cancer stem cell (CSC) generation and drug resistance in many cancers. Targeting EMT may be promising to benefit cancer treatment. Our previous high throughput screening study has identified a potential EMT inhibitor (namely, C150) in pancreatic cancer cells. Here, we sought to investigate the activities of C150 in inhibiting pancreatic cancer cell invasion, CSCs and tumor growth in mice, and its mechanisms of EMT inhibition.MethodsCell viability was detected by MTT assay. Cell migration/invasion was detected by Boyden chamber trans‐well migration‐invasion assay, wound scratch assay and a 3‐dimension (3D) cell invasion assay. Tumor sphere formation in stem cell medium was used to indicate CSC inhibition. In vivo tumor growth was evaluated in an orthotopic mouse model of pancreatic cancer. EMT marker proteins were examined by western blot.ResultsC150 exhibited a well‐separated cytotoxicity between pancreatic cancer cells and non‐cancerous cells, with IC50 values of 1~2.5 μM in multiple pancreatic cancer cell lines and >25 μM in a non‐cancerous pancreatic epithelial cell line (hTERT‐HPNE). C150 was found to significantly inhibit PANC‐1 cell invasion in the 3‐dimension (3D) cell invasion model, the wound scratching assay and the Boyden Chamber trans‐well migration‐invasion assay. Pancreatic tumor sphere formation was strongly inhibited by C150 at concentrations as low as 0.4 μM in PANC‐1 cells and 0.05 μM in MiaPaCa‐2 cells, indicating preferable inhibition of pancreatic cancer stem cells. In the orthotopic pancreatic cancer mouse model, C150 significantly reduced tumor growth at the dose of 15 mg/kg, 3x weekly IP injection. C150 decreased the mesenchymal marker N‐cadherin while increased epithelial markers of ZO‐1 and Claudin‐1. Further studies revealed that C150 significantly reduced total and nuclear levels of Snail and β‐Catenin, two important transcription factors that contribute to EMT and cancer stem cells in pancreatic cancer.ConclusionThe novel compound C150 inhibited pancreatic cancer cell migration, invasion, tumor sphere formation and in vivo tumor growth in mice, by inhibiting EMT through Snail and β‐Catenin down‐regulation. The study of detailed mechanisms on how C150 reduced Snail and β‐Catenin protein levels is currently ongoing.Support or Funding InformationInstitutional Development Award (IDeA) of NIH and State of Kansas Grant # 8P30GM103495, KU Cancer Center Lied Basic Science grant 2018This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hypersensitivity to Cisplatin and Gentamycin Induced Nephrotoxicity in Mice with Decreased Expression of NADPH‐Cytochrome P450 Reductase
Authors: Ding Liang , Zhang Qingyu , Ding Xinxin 
Published: June 2021

Cytochrome P450 reductase (CPR or POR) is an obligatory redox partner for all microsomal P450 enzymes. A large, global decrease in POR expression, in a Cpr‐low mouse model, was previously found to lead to altered sex steroid hormone homeostasis, female infertility, and decreased capacity for xenobiotic metabolism. The aim of this study was to further characterize the Cpr‐low mouse model, in order to identify additional phenotypes that may reveal vulnerability in people with low POR expression. Here, we report differences between wild‐type and Cpr‐low mice in their sensitivities to drug‐induced toxicity in the renal proximal tubules. We studied two drugs, the anticancer drug cisplatin and the antibiotic drug gentamycin, which are known as renal toxicants, but are not P450 substrates. Toxicity was assessed by blood urea nitrogen (BUN) levels and histopathology of the kidney. At drug doses that do not cause renal toxicity in wild‐type mice, cisplatin caused significant increases in BUN levels over vehicle control group in both male and female Cpr‐low mice, whereas gentamycin caused significant increases in BUN levels over vehicle control group in male but not female Cpr‐low mice. Corresponding histopathological changes, including appearance of extensive proximal tubule vacuolization, were found in the kidneys of drug‐treated Cpr‐low mice. These findings suggest that POR expression level could be a risk factor for drug‐induced nephrotoxicity in patients.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Pannexin 1 Channels Reduces Tumorigenic Properties of Melanoma
Authors: Penuela Silvia , Freeman Taylor Jessica , Sayedyahossein Samar , Johnston Danielle , Sanchez‐Pupo Rafael Enrique , O'Donnell Brooke Leone , Huang Kenneth , Lakhani Zameena , Nouri‐Nejad Daniel , Barr Kevin J , Harland Luke , Latosinsky Steven , Grant Aaron 
Published: June 2021

Melanoma is the deadliest skin cancer and its incidence is still on the rise. Despite new advances with immunotherapy drugs, nearly 70% of patients still do not respond to treatments, underscoring the importance of finding new treatment targets and combination therapies. Pannexin 1 (PANX1 human; Panx1 mouse) is a channel‐forming glycoprotein expressed in many mammalian organs and tissues including the skin at early stages of development. Pannexin 1 channels allow the passage of ions and important signaling molecules up to 1 kDa, including ATP and other metabolites. We previously found that a knockdown of Panx1 resulted in reduced tumorigenic properties of mouse melanoma cell lines in vitro. In this study, we found that PANX1 is highly expressed in human melanoma tumors compared to normal skin. Using Western blot and immunohistochemistry, we showed that high PANX1 levels are present in melanoma tumors at all disease stages, as well as in patient‐derived cells and established melanoma cell lines. We demonstrated that inhibiting PANX1 function using shRNA or channel blockers, such as Carbenoxolone (CBX) and Probenecid (PBN), significantly reduced cell growth and cell migration, and substantially increased the production of melanin (used as a differentiation marker) in A375‐P and A375‐MA2 human melanoma cells. In addition, treatment with CBX or PBN in A375‐MA2 cells xenografted onto the chorioallantoic membrane of a chicken embryo model, significantly reduced primary melanoma tumor weight as well as tumor invasion. Blocking PANX1 channels using PBN reduced ATP release in A375‐P melanoma cells, suggesting a role for PANX1 in regulating the tumor microenvironment through purinergic signaling at the cell surface. In addition, our findings from cell‐surface biotinylation assays indicate that there is also a significant intracellular pool of PANX1 in melanoma cells that likely modulates signaling pathways such as Wnt/β‐catenin, since β‐catenin levels are significantly decreased upon shRNA knockdown of PANX1 in melanoma cells. Collectively, our findings suggest that PANX1 is a novel modulator of tumorigenic properties in human melanoma cells, contributing to signaling pathways that regulate melanoma progression and highlights PANX1 as a potential target for therapeutic intervention.Support or Funding InformationFunded by a Canadian Institutes of Health Research (CIHR) Project Grant to Silvia PenuelaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A small, synthetic glycosaminoglycan mimetic as an inhibitor of human heparanase
Authors: Sistla Jyothi C. , Afosah Daniel K , Morla Shravan , Patel Nirmita J. , Patel Bhaumik B. , Desai Umesh R. 
Published: June 2021

Human heparanase (HPSE) is an endo‐b‐D‐glucuronidase that cleaves glycosaminoglycan (GAG) chains of heparan sulfate proteoglycans (HSPGs) present on cell surfaces and in the extracellular matrix (ECM). HPSE is upregulated in many types of cancers and promotes an aggressive tumor growth leading to metastasis. HPSE is also involved in various other pathologies such as inflammation, thrombosis, atherosclerosis and kidney dysfunction. Because of these important roles, HPSE is a promising target for the development of a novel therapeutic. In fact, several HS mimetics including PI‐88, SST000, PG545, and M402 are being pursued in clinical trials. Yet, each of these is saccharide mimetics of HS, which are either polymeric, and therefore heterogeneous, or are very difficult to prepare. We have pursued the idea that small, synthetic agents called Non‐Saccharide Glycosaminoglycan Mimetics (NSGMs) would modulate pathological responses such cancer, coagulation, and inflammation in which GAGs play important roles. We hypothesized that an optimal small, synthetic NSGM that potently binds to HPSE would inhibit its enzymatic activity and lead to a novel, anti‐angiogenesis agent. Therefore, we screened an in‐house library 70 NSGMs and identified 35 agents that displayed reasonable HPSE inhibition. Detailed studies using a one‐step FRET assay as well as UPLC‐MS/MS assay showed that an NSGM labeled as G2.2 exhibited high anti‐HPSE potency of 3.2±0.3 μM. G2.2 is synthesized in five simple steps in high overall yields (>30%) from readily available natural product quercetin. G2.2 significantly blocked the migration and invasion of HT‐29 spheroid cells compared to vehicle‐treated control cells. The highly promising anti‐HPSE agent is currently being investigated in animal models of angiogenesis.Support or Funding InformationP‐01 HL107152This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antioxidant α‐mangostin downregulates ATP‐binding cassette transporters in HuT78 cells
Authors: Jungsuwadee Paiboon 
Published: June 2021

Sézary syndrome (SS) is an aggressive form of cancer called cutaneous T‐cell lymphoma (CTCL). Patients with SS carry poor prognosis, with a median survival of approximately 40 months after diagnosis and a 5‐year survival rate below 40%. The offending neoplastic T cells for SS known as Sézary cells are aggressive and rapid growing. They are refractory to chemotherapy, and have been posting the biggest challenge for the treatment of SS. The principal mechanism by which the Sézary cells develop resistance to chemotherapeutic agents is through the overexpression of ATP‐binding cassette (ABC) transporter proteins such as P‐glycoprotein (P‐gp), multidrug‐resistance associated protein 1 (MRP1), and breast cancer resistance protein (BCRP). The expressions of P‐gp, MRP1 and BCRP genes appear to be regulated, in part, by nuclear factor E2‐related factor 2 (Nrf2), a transcriptional regulator of cellular resistance to oxidants. α‐Mangostin (α‐MGT) is a xanthone derivative that possesses antioxidant and cytotoxic activities in various types of cancers. This study is undertaken to support the hypothesis that through its antioxidant activity, α‐MGT mediates downregulation of ABC transporters expression in HuT78, the cell line isolated from a Sézary patient. The results revealed that HuT78 cells expressed P‐gp, MRP1 and BCRP at various frequencies, ranging from 31% to 78% where the expression level of MRP1 was the highest followed by that of BCRP and P‐gp. Upon exposure of HuT78 cells to 12 μM α‐MGT for 24 hours, the expression of MRP1 was decreased by 50%, BCRP was slightly downregulated, and the P‐gp was undetectable. Our preliminary data suggested that α‐MGT may be beneficial in reversing the resistant phenotype of Sézary cells for chemotherapy.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A novel superoxide dismutase mimetic sensitizes lung cancer cells to radiation and protects normal cells
Authors: Alzarie Yasmin , Badary Osama , Mansoor Fatma , El‐Mahdy Mohamed , Zweier Jay 
Published: June 2021

Lung cancer remains the leading cause of cancer mortality worldwide, and ionizing radiation (IR) is considered one of the most successful strategies to treat such an aggressive disease. However, the large doses of IR required to effectively kill cancer cells induce significant damage to normal cells, leading to treatment failure. In the present study, we investigated the effects of GC4419, a newly developed superoxide dismutase mimetic, alone and in combination with different doses of IR on cell proliferation of normal and non‐small cell lung cancer; the molecular mechanisms of such effects were also studied. Our aim was to investigate the ability of GC4419 to prevent or minimize radiotherapy‐induced adverse effects to normal cells and to enhance the efficacy of low doses of IR to kill cancer cells. Our results revealed that treatment with GC4419 alone inhibited cancer cells proliferation without affecting the viability of normal cells. Notably, combination treatment of GC4419 and IR inhibited cell proliferation of cancer cells more significantly, when compared to IR mono therapy. DNA fragmentation and caspase cleavage were significantly increased following combined GC4419 and IR treatment, indicating activation of apoptosis pathways. Furthermore, GC4419 or IR alone increased the ratio of Bax/Bcl‐2 protein expression, indicating induction of apoptosis. Such an effect was significantly increased following the combined therapy. In conclusion, our study demonstrates that GC4419 can be used as an effective adjuvant to radiation therapy to enhance the efficacy of low dose IR and to protect normal cells against radiation‐induced damage.Support or Funding InformationNIH grants # HL 135648, HL 131941, and EB 016096 to Jay L. ZweierThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.