bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Novel Small Molecule Kinase Antagonists of the Prolactin Receptor In Breast Cancer
Authors: Bakshi Chinmay 
Published: June 2021

Breast cancer remains to be a prevalent malignancy, as an estimated 231,840 women were diagnosed in the US in 2015. Furthermore, cancer cells have developed chemoresistance, necessitating the pursuit of innovative targets. Prolactin (PRL) acting via the prolactin receptor (PRLR) is an anti‐apoptotic hormone in breast cancer. Extensive research indicates that PRLR activation in breast cancer is associated with enhanced tumor growth, invasiveness, metastasis, and resistance to chemotherapy.To identify novel antagonists of the PRLR, a 340,000 small molecule library was scanned using high throughput screening and computational modeling techniques (in‐silico docking) to observe binding potential to the PRLR. Kd and IC50 values were found using cell based assays for testing ligand binding efficiency in T47D and Ba/F3 cell lines. 2 molecules, SMI 1 and SMI 6, were identified as potential inhibitors of the PRLR. Inhibitor cytotoxicity was tested using an LDH assay, utilizing a 468 cancer cell line, to ensure drug viability as a potential treatment. An invasion assay was produced to test cell invasion when exposed to 1nM prolactin in a 468 cell line. Immunoprecipitation and western blotting were used to observe downregulation of the PRLR signaling pathway. Cancer cell proliferation after inhibitor exposure was calculated using flow cytometry.Results indicated SMI 1 and 6 had minimal toxicity, ensuring both molecules' capabilities to be therapeutic agents. Furthermore, both inhibitors reduced cell invasion, and cell proliferation when exposed to prolactin, compared to exposure to prolactin only. Additionally, the inhibitors also appear to downregulate the STAT‐5 signaling pathway. Thus, both these small molecule antagonists hold great promise in targeting the prolactin receptor as potential cancer therapeutic drugs of the future.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structure‐Activity Relationship of Novel Compounds Based on Tamoxifen with Proposed Sigma‐2 Receptor Activity
Authors: Stang Christopher R.T. , Inbody Laura R. , Orahood Cassandra B. , Dick Matt J. , Eichel Amanda M. , Mohamed Ahmed H. , Faler Ashton D. , Zaibek Ann E. , Kinder David H. , Schneider Ryan A. 
Published: June 2021

Sigma‐2 receptors are overexpressed in several different types of human cancer cells and are regarded as potential therapeutic targets. Ligands for sigma‐2 receptors have been shown to elicit anticancer properties including growth arrest and apoptosis. While much research has been devoted to designing sigma‐2 receptor ligands for therapeutic benefits, none have stood up to rigorous testing and clinical trials needed for approval in human patients. In an attempt to provide more literature on the potential usefulness of sigma‐2 receptor ligands for anticancer activity, we synthesized a series of analogs based upon the known sigma‐2 receptor ligand tamoxifen that lack estrogen receptor binding properties.We previously reported on the first molecule in this series, SP‐1, with IC50 activity ranging from 30–60 μM against several dissimilar cell lines. To further elucidate structurally important regions of the molecule and ascertain a structure‐activity relationship (SAR) for anticancer activity, six additional molecules were synthesized and tested. These molecules included modifications to both the terminal phenyl and nitrogen moieties of SP‐1 (N,N‐dimethyl‐2‐(4‐(1‐(p‐tolyl) vinyl)phenoxy)ethan‐1‐amine). The XTT cell viability assay was then used to screen these compounds for anticancer activity in four diverse human cancer cell lines including A‐172 human glioblastoma, A375 human malignant melanoma, HT‐29 human colorectal adenocarcinoma, and MCF7 human breast adenocarcinoma. Interestingly, hydration of the ethylene moiety attached to the carbon linker between the two phenyl rings markedly reduced anticancer activity of SP‐2, SP‐3, SP‐7 and SP‐8. Substantial anticancer activity was not observed at concentrations less than 90 μM regardless of structural modifications made to the periphery of these molecules. However, SP‐5 and SP‐6, which conserved the nonpolar central linker present in SP‐1, demonstrated moderately potent anticancer activity against the four cancer cell lines. SP‐5, which only differs from SP‐1 by the addition of one carbon to the peripheral phenyl ring, was found to have nearly the same potency as SP‐1. However, SP‐6, which includes the addition of a morpholine ring to the periphery of the molecule, demonstrated slightly less potent activity than that of SP‐1 and SP‐5. However IC50 values were still found to range from 45–90 μM in all four cells lines. While the changes made to these analogs failed to improve the efficacy of our parent molecule SP‐1, the information regarding the SAR will help guide the synthesis of further analogs. Additional studies investigating the mechanisms of death and putative targets are ongoing for these lead molecules.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Curcumin Attenuates Expression of Adhesion Molecules by IL‐1 β in HUVECs
Authors: Chang Weng‐Cheng , Yu Ya‐Mei , Cheng An‐Chin 
Published: June 2021

Atherosclerosis, a progressive pathological disorder inducing cardiovascular and cerebrovascular diseases, is a leading cause of human morbidity and mortality worldwide. The expression of cell adhesion molecules such as intercellular adhesion molecule (ICAM‐1), vascular cell adhesion molecule‐1 (VCAM‐1), and adhesion molecule‐1 (E‐selectin) by the endothelium and the attachment of monocytes to endothelium may play a pivotal role in the early atherogenic process. Curcumin is a polyphenol obtained from the root of the turmeric (Curcuma longa), and is the main component of curry powder for giving a specific flavor and yellow color. Previous studies indicated that curcumin exhibits a variety of pharmacological effects including anti‐inflammatory, antitumor, antioxidant, anti‐hypertensive, antiviral, anti‐obesity, anti‐infectious activities and wound‐healing properties. In this study, efforts were made to identify curcumin may aid in the prevention of monocyte‐endothelial cell adhesion induced atherosclerosis. Our results indicated that curcumin could inhibit the expression of adhesion molecules, including ICAM‐1, VCAM‐1, and E‐selectin, and futher decrease the adhesiveness to a human monocytic cell line (U937) in human umbilical vein endothelial cells (HUVECs). These results demonstrated that curcumin has inhibitory effect on proanthersclerotic mechanism in vitro.Support or Funding InformationThis study was supported by NSC 100‐2313‐B‐309‐003 to A‐C Cheng and NSC 95‐2815‐C‐039‐019‐B to Y‐M Yu.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of STAT3 phosphorylation by curcumin partly reduces adhesion molecule expression in vascular endothelial cell
Authors: Ahn Hee Yul , Cho Young Suk , Sankaralingam Sowndramalingam 
Published: June 2021

OBJECTIVESWe investigated whether curcumin, an ingredient of turmeric, can suppress the expression of intercellular adhesion molecule 1 (ICAM‐1) and vascular cell adhesion molecule 1 (VCAM‐1) in human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS), and the underlying mechanisms.MATERIALS AND METHODOLOGYThe levels of adhesion molecules, activities of nuclear factor kappaB (NF‐kB) and signal transducer and activator of transcription 3 (STAT3) were determined using Western blot analysis. The effect of curcumin on monocyte binding to endothelial cells was determined using THP‐1 monocyte (human acute monocytic leukemia cell line) adhesion assay.RESULTSCurcumin prevented the LPS‐mediated increase in ICAM‐1 and VCAM‐1 expression in HUVEC in a dose‐dependent manner. Curcumin did not affect LPS induced degradation of IkappaBα (IkBα). On the other hand, curcumin reduced the LPS‐induced STAT3 phosphorylation. Curcumin significantly reduced LPS‐mediated THP‐1 monocyte adhesion to HUVEC. Furthermore, in C57BL/6 mice, curcumin prevented the LPS induced increase in aortic ICAM‐1 and VCAM‐1 expression.CONCLUSIONSCurcumin prevented LPS‐induced ICAM–1 and VCAM–1 expression on vascular endothelial cells associated with inhibition of STAT3 phosphorylation.Support or Funding InformationThis work was supported by the research grants of the Chungbuk National University in 2014.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FAK maintenance of endothelial mechanotransduction controls epigenetic repression of KLF2 and S1PR1 transcription
Authors: Akhter Md Zahid , Yazbeck Pascal , Srivastava Nitya N. , Tauseef Mohammad , Tennes‐Schmidt Tracy , Lenzini Stephen B. , Shin Jae‐Won , Mehta Dolly 
Published: June 2021

Sphingosine‐1‐phosphate (S1P) activation of sphingosine‐1‐phosphate receptor 1 (S1PR1) in endothelial cells (ECs) strengthens the endothelial barrier and prevents lung injury. However mechanisms regulating EC‐S1PR1 expression remains unclear. Recently, we showed that endothelial‐specific deletion of focal adhesion kinase (FAK) in mice severely disrupted lung vascular barrier function. Microarray analysis of FAK depleted cells showed a marked reduction in S1PR1 expression. Thus, we tested the hypothesis that impaired S1PR1 synthesis is responsible for defective endothelial barrier repair in EC‐FAK−/− mice. We confirmed that FAK deletion markedly reduced S1PR1 expression both at the mRNA and protein level in lungs and ECs whereas the expression of S1PR2 and S1PR3 and S1P generating enzymes, sphingosine kinases (SPHK1 and SPHK2) was not reduced. Consistently, FAK depleted ECs and mice conditionally lacking FAK in pulmonary ECs failed to anneal adherens junctions and showed impairment of endothelial barrier function under basal condition and even after S1P application. Rescuing S1PR1 expression in FAK null ECs restored barrier function in the pulmonary vessels of EC‐FAK null mice. Next, we screened for potential transcription factors binding to the S1PR1 promoter which included KLF2, Sox2, FoxP1, GATA1, GATA4 and ERG1. Intriguingly, we observed that rescuing KLF2 expression in FAK depleted ECs restored S1PR1 expression and endothelial barrier function to the levels observed in control cells. In other studies, we observed that KLF2 increased S1PR1 promoter activity while the S1PR1 cDNA lacking KLF2 binding sites failed to increase S1PR1 promoter activity, demonstrating that KLF2 can promote endothelial barrier function in the absence of EC‐FAK by transducing S1PR1. Because FAK maintains intracellular tension by balancing the activities of RhoA and Rac1 GTPases and KLF2 mRNA expression is regulated by methylation of CpG islands, we next address the possible role of increased cellular tension and DNA methyltransferases (DNMT) in suppressing KLF2 gene transcription and thereby S1PR1 expression in EC‐FAK null mice. We found that DNMT activity was elevated by 3‐fold in FAK depleted ECs in a RhoA‐dependent manner. Intriguingly, we found that depletion of FAK increased EC‐stiffness, which was associated with hypermethylation of KLF2 by DNMT3a. Interestingly, inhibition of RhoA in FAK depleted cells restored KLF2 and S1PR1 expression. Thus, our data describes the novel role of FAK in suppressing cellular tension driven‐epigenetic modification of KLF2 and S1PR1 and thereby vascular integrity.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiac‐specific Overexpression of MiR‐133a Decreases Pyroptosis in Ins2+/− T1DM Mice Heart
Authors: Kambis Tyler N. , Yadav Santosh K. , Mishra Paras K. 
Published: June 2021

Increased pyroptosis (a caspase‐1‐dependent cell death) and decreased miR‐133a (the most abundant cardioprotective miRNA in the human heart) are documented in the diabetic mellitus (DM) heart. However, it is unclear whether the reduced miR‐133a in DM hearts induces cardiac pyroptosis. We hypothesized that lack of miR‐133a in the diabetic heart induces pyroptosis, which is suppressed by the overexpression of miR‐133a. To test the hypothesis, we created a novel diabetic mice strain where miR‐133a is overexpressed only in the heart (Akita/miR‐133aTg), by crossbreeding Ins2+/− Akita (spontaneous, genetic T1DM) male with cardiac‐specific miR‐133a transgenic (miR‐133aTg) female mice. We have used sibling Ins2+/+ mice as WT for Ins2+/− Akita mice. In the four groups (WT, Akita, Akita/miR‐133aTg and miR‐133aTg) of fourteen‐week male mice, we measured the levels of pyroptosis markers NLRP3 (involved in the inflammasome complex formation that is required for pyroptosis initiation), caspase‐1 (the key enzyme for pyroptosis), and interleukine‐1β (IL‐1β) (a pro‐inflammatory cytokine that is involved in pyroptosis) in the heart. The densitometric analyses of Western blots revealed that protein levels of pyroptosis markers are upregulated in Akita hearts, whereas the overexpression of miR‐133a blunts this upregulation. The protein expression of different pyroptosis markers in the above‐mentioned four groups of hearts were; NLRP3: WT 0.04 ± 0.0, Akita 0.09 ± 0.1, Akita/miR‐133aTg 0.03 ± 0.0, miR‐133aTg 0.01 ± 0.0; caspase 1: WT 0.7 ± 0.1, Akita 1.0 ± 0.1, Akita/miR‐133aTg 0.2 ± 0.0, miR‐133aTg 0.1 ± 0.0, and IL‐1β: WT 1.9 ± 0.6, Akita 2.1 ± 0.3, Akita/miR‐133aTg 1.1 ± 0.0, miR‐133aTg 0.8 ± 0.1 (the standard deviation values are rounded to one decimal point). In conclusion, this is the first report showing that cardiac‐specific overexpression of miR‐133a mitigates DM‐induced pyroptosis in the heart.Support or Funding InformationDepartment of Cellular and Integrative Physiology, University of Nebraska Medical CenterThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Rbfox1 regulates proteasomal activity and K48‐linked ubiquitination
Authors: Umar Sadiq , vanBerlo Jop 
Published: June 2021

Rbfox1 is important to maintain cardiac function through regulation of alternative splicing and expression of calcium handling proteins. The heart recycles proteins and degrades them either via the ubiquitin proteasome system or through autophagy. The role of Rbfox1 in post‐translational modification (PTM) and proteasomal degradation is not known. To assess a potential role for Rbfox1 in regulating PTM and proteasomal degradation, we deleted Rbfox1 in cardiomyocytes, either by Rbfox1 gene‐deletion mice (cRbfox1−/−) or by transfecting cultured neonatal rat cardiomyocytes with Rbfox1siRNA. Cardiomyocyte specific deletion of Rbfox1 was confirmed by Western blotting and immunohistochemistry. cRbfox1−/− mice showed an overall decrease in expression of SUMO‐1 and K63‐ubiquitin conjugated proteins while K48‐Ubiquitin expression was increased. These results were confirmed in cardiomyocytes transfected with Rbfox1 siRNA, suggesting a direct role for Rbfox1 in regulating sumoylation and ubiquitination. Moreover, we measured the 20S proteasome activity and found an increase in proteasomal activity both in cRbfox1−/− mouse as well as in Rbfox1siRNA transfected cardiomyocytes. Since protein abundance is a balance between protein production and degradation, we further investigated protein synthesis. We transfected cardiomyocytes with siRNA against Rbfox1 and treated them with puromycin. Cell lysates were probed with a puromycin antibody to measure overall protein synthesis. Our results showed significant deficiency in total protein synthesis in Rbfox1 siRNA as compared with control. In conclusion, Rbfox1 is important in maintaining protein homeostasis, by controlling their production and degradation. These alterations in critical cellular processes likely predispose the heart to cardiac dysfunction.Support or Funding InformationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Functional anatomy of the masticatory muscles in a caviomorph rodent, Cavia porcellus
Authors: Druzinsky Robert E. , Ulm Ashley , Vinyard Christopher J. , Iriarte‐Diaz Jose 
Published: June 2021

Guinea pigs (Cavia porcellus) and other caviomorph rodents are hystricognaths. In hystricognaths, there is a groove for the pars reflexa of the superficial masseter in the ventral border of the mandible (Druzinsky, 2007). In the guinea pig (C. porcellus) the tendon of the pars reflexa has a dense connective tissue sesamoid that permits the tendon to glide through the groove. These South American hystricognaths are also hystricomorphs. The zygomatico‐mandibularis muscle originates on the snout and converges on a tendon that runs posteriorly through the infraorbital foramen. The tendon runs over the bony root of the zygomatic arch and turns inferiorly, to insert at the anterior end of the masseteric groove. A second sesamoid is found in this tendon, which allows the tendon to glide over the root of the zygomatic arch. Each of these elaborations of the masticatory muscles is a pinnate muscle that curves around a bony structure, acting as a pulley, and permitting the muscle to be longer than similar muscles in other mammals.To analyze the mechanics of the caviomorph masticatory apparatus we are creating a 3D model of the skull, jaw, and masticatory muscles through contrast enhanced micro‐CT. The head of an adult guinea pig was soaked in a solution of called iodine potassium iodide (I2KI) for several weeks and then scanned at Northeast Ohio College of Medicine using the Scanco vivaCT micro‐CT. The Materialize Mimics® Innovation Suite will be used to segment the bones, teeth, and muscles to capture the detailed morphology. Modeling of length‐tension relationships and mechanical advantage suggests that the pulley mechanisms of the masseter complex permit these muscles to be long, which allows them to have long excursions and produce significant forces at large gapes.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enoxaparins Derived from Ovine (Sheep) Tissues are Biosimilar to Branded Enoxaparin of Porcine (Pig) Origin
Authors: Kouta Ahmed , Hoppensteadt Debra , Abro Schuharazad , Clarizio Destini , Jeske Walter , Yao Yiming , Fareed Jawed 
Published: June 2021

Background and PurposeLow molecular weight heparins (LMWH) are widely used for the management of thrombosis. Enoxaparin represents one of the most widely used LMWH, obtained by depolymerizing porcine (pig) mucosal tissues. Recently, heparin from sheep mucosal tissue has also become available which exhibits a comparable anticoagulant profile to porcine heparin. The purpose of this study is to compare enoxaparin from ovine mucosa with branded enoxaparin.Materials and MethodsFour batches of enoxaparin were commercially obtained (SanofiAventis, Bridgewater, NJ, USA). Four ovine enoxaparins were provided by Ronnsi Pharmaceutical Co., China. These enoxaparins were compared for the anticoagulant (ACT, aPTT, TT), antiprotease (Anti‐Xa, Anti‐IIa), and their potency was cross‐referenced using NIBSC LMWH standard in an amidolytic Anti‐Xa method (Hyphen Biomedical, Paris, France). Molecular profile studies were carried out as specified by U.S. FDA. Protamine titration studies were carried out for both preparations using various tests. The relative interactions of these enoxaparins with HIT antibodies were also investigated using HIT antibodies obtained from patients employing platelet aggregation studies. Thrombin generation inhibition studies were also carried out using a fluorogenic substarte method (Technoclone, Austria)ResultsThe molecular profile of ovine and porcine enoxaparins were comparable and ranged from 4.2–4.5 kDA (p<0.05). The anticoagulant activities in all the assays were also comparable for the ovine and porcine enoxaparins. In the anti‐Xa assays, the NIBSC cross‐referenced potency of several batches of porcine enoxaparin ranged from 98–110 U/mg (mean=102+/−4) whereas the ovine enoxaparin exhibited 102–112U/mg (mean=106+/−6) potency (p<0.05). In the anti‐II assays the potency of the ovine enoxaparins ranged from 25–30 u/ml (mean 28 ± 3) and porcine enoxaparin were 28–35 (mean 31 ± 3). In the protamine titration curves, both enoxaparin preparations provided superimposable results. Similarly, in the HIT antibody assays, the behavior of ovine and porcine enoxaparin was identical.ConclusionsThese studies demonstrated that ovine mucosal tissue‐derived enoxaparin exhibits similar molecular and anticoagulant profiles. In the NIBSC cross‐referenced potency assay, the potency of these preparations were similar. Thus, ovine enoxaparins exhibit comparable profiles when compared to their porcine counterparts and are expected to provide similar results in various clinical indications.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alkaloid Fraction of Alstonia Boonei Stem Bark Demonstrate Antiobesity Potential on High Fat Diet Induced Obesity in Male Sprague‐Dawley Rats
Authors: Anyanwu Gabriel Oluwabunmi , Onyeneke Chukwu Eusebius , Iqbal Jamshed , Qamar Misbah ud Din , Khan Shafi Ullah , Rehman Sabi ur , Rauf Khalid , Rehman Nisar ur 
Published: June 2021

The aim of this study was to investigate the anti‐obesity potential of alkaloid fraction of Alstonia boonei stem bark on high fat diet (HFD) induced obesity in male Sprague‐dawley rats.Preparation of neutral alkaloid fraction (NAF) was done through solvent partitioning of the methanol extract of Alstonia boonei stem bark, and NAF was subjected to LCMS‐MS analysis. To induce obesity, rats were fed with HFD (60% fat content) for 12 weeks preceding oral treatment with 125 and 250 mg/kg body weight of NAF for 4 weeks. Experimental procedures involving rats followed the European Community guidelines (EEC Directive of 1986; 86/609/EEC) for animal use.The major compounds found in NAF includes: Ajmalicine, 16‐Methoxy‐2,3‐dihydro‐3‐hydroxytabersonine, Vincamine, Aspidofractine, 3‐Hydroxyquinidine, 16‐Methoxytabersonine, Mitraphylline, 18‐Hydroxyyohimbine and Catharanthine. NAF significantly (p < 0.05) decreased body weight, white adipose tissue, triglycerides, total cholesterol, low density lipoprotein cholesterol, very low density lipoprotein cholesterol, leptin, insulin and dopamine levels, but increased significantly (p < 0.05) the high density lipoprotein cholesterol, cortisol, serotonin and noradrenaline levels in the treated groups compared to HFD control. However, there were no significant (p > 0.05) changes in food intake, blood glucose, thyroxine, triiodothyronine and thyroid stimulating hormone levels in animals given NAF when compared to HFD control. The photomicrograph of liver of the rats treated with NAF showed slight fat changes unlike that of the HFD obese controls which showed presence of numerous prominent fat deposits on the livers.The findings in this study suggests that the alkaloid fraction of A. boonei possess antiobesity properties as indicated in improved lipid profile, leptin sensitivity, and decreased body weight of animals.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.