bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Promiscuous Biotin Ligase as a Global Strategy to Interrogate the Protein Interactome of Human ABCC4
Authors: Looi‐Somoye Remi , Wijaya Juwina , Peng Junmin , Schuetz John 
Published: June 2021

ATP‐Binding Cassette transporter sub‐family C member 4, ABCC4, is a plasma membrane transporter with the ability to export prostaglandins and cancer chemotherapeutics. In acute myeloid leukemia (AML), ABCC4 interacts with a binding protein through its PDZ motif1. This interaction with a highly expressed PDZ protein, a protein which correlated with worse overall survival for AML patients, affects ABCC4 plasma membrane localization and retention, leading to profound effects on its function and drug resistance. The intense focus on ABCC4's PDZ motif as a locus for protein interaction has left other potential protein interactions that might regulate ABCC4 function largely unexplored. Little is actually known about how ABCC4 traffics to and from the plasma membrane. In the present study, we developed a global approach to not just identify potential interacting proteins, but also determine the proteins required for ABCC4 trafficking and internalization. We used a promiscuous biotin ligase, BioID2, which covalently attaches a biotin moiety to nearby proteins in the presence of biotin. Fusing BioID2 to the N‐terminus of ABCC4 did not alter its glycosylation, plasma membrane localization, or function. Upon addition of biotin, the BioID2‐ABCC4 fusion protein was capable of biotinylating itself as well as ABCC4 proximal and interacting proteins. We then developed an immunoprecipitation and elution protocol and strategy to specifically identify proteins biotinylated by the BioID2‐ABCC4 fusion protein. With successful immunoprecipitation of biotinylated proteins and identification by mass spectrometry, proteins within the vicinity of ABCC4, from its genesis at the endoplasmic reticulum to its final destination at the plasma membrane, will be identified. At the same time, proteins involved in ABCC4 recycling to degradation will also be discovered. Overall, this study will provide unique insights into the mechanisms of ABCC4 trafficking and identify proteins that will allow for modulation of ABCC4 level. This will be beneficial for malignancies associated with high ABCC4 expression, including neuroblastoma, prostate, non‐small cell lung, and endometrial cancers.Support or Funding InformationThis work was supported by NIH and ALSAC.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Polyamine pathway as a player in breast cancer cell proliferation in diabetic conditions
Authors: Capellen Caleb , Ortega Jose , Morwitzer M. Jane , Dunworth Matthew , Casero Robert A. , Chandra Surabhi 
Published: June 2021

Diabetes/hyperglycemia has been known to increase cancer cell proliferation, leading to metastasis and even mortality. High blood sugar is also known to reduce the effects of cancer therapeutics. Polyamines (spermine, spermidine, and putrescine) which are involved in several cellular processes such as cell growth, replication, and transcription, have been shown to be elevated in certain cancers including colon cancer and skin cancer. We hypothesized that polyamines are involved with proliferation of breast cancer cells in diabetic conditions. Cell lines used in this study included MCF‐7 (early stage breast cancer), MDA‐MB‐231 (late stage breast cancer), and MCF‐10A (normal breast epithelial cells). Cells were treated with normal glucose (NG, 5mM) or high glucose (HG, 25 mM) in the presence/absence of polyamine pathway inhibitors. Both MDA‐MB‐231 and MCF‐10A cells showed significant proliferation with HG treatment after 72 hr, which was not observed in MCF‐7 cells. Simultaneous treatment with an ornithine decarboxylase (polyamine synthesis enzyme) inhibitor, difluoromethylornithine (DFMO) could prevent this effect. Interestingly, while spermine levels were not altered with HG treatment in breast cancer cells (MCF‐7 and MDA‐MB‐231), they were significantly increased in MCF‐10A cells. This is possibly due to the fact that cancer cells have endogenously high levels of polyamines, such that further increase is prevented to avoid toxicity from polyamines. Nevertheless, DFMO was able to decrease spermine and spermidine levels induced with glucose treatments, suggesting a role of polyamine pathway in regulating growth. Putrescine levels were undetectable in all samples. In conclusion, polyamine pathway is involved with HG mediated normal breast epithelial cell and breast cancer cell proliferation. Future studies will involve further analysis of the mediators which regulate polyamine production.Support or Funding InformationUNK Faculty Start‐up Funds (SC); Undergraduate Research Fellowship Grant (CC and JO)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Down Regulation of Add3 in Astrocytes Disrupts the Actin Cytoskeleton in Association with Decreasing Small Molecule Uptake and May Contribute to Cognitive Deficits in FHH rats
Authors: Travis Olivia Kristell , Wang Shaoxun , He Xiaochen , Roman Richard J. , Fan Fan 
Published: June 2021

We previously reported that the expression of Add3 is decreased in various tissues, including brain and cerebral vessels, and further decreases with age in FHH rats. More recently, we demonstrated that Add3 dysfunction contributes to cerebral vascular impairments in FHH rats, and they exhibit BBB leakage and neurodegeneration after the development of hypertension. In addition, Aβ protein expression in the brain in FHH rats is increased as early as 8 weeks of age. Reactive astrogliosis is a common finding in neurodegenerative diseases and activated astrocytes promote neurodegeneration. Moreover, astrocyte dysfunction affects Aβ clearance and Aβ accumulation is a characteristic biomarker of Alzheimer's disease. To further explore whether astrocytes contribute to neurodegeneration and cognitive impairments in FHH rats, we knocked down Add3 expression using Add3 Dicer‐substrate RNAi (DsiRNA) in human astrocytes and found that GFAP expression was markedly enhanced in comparison to scrambled siRNA treated cells. F‐actin distribution was significantly reduced in the Add3 DsiRNA treated cells (19.3 ± 6.7%, n = 6) when compared to the control cells (34.9 ± 3.0%, n = 6). In addition, the Add3 DsiRNA treated cells took up less fluorescence‐labeled dextran (10 kD) than the control cells suggesting the capability of uptake of small molecule, such as Aβ may be diminished. We found that DOCA/salt induced hypertensive FHH rats exhibit a reduction in neuronal density and size in the hippocampus. This was associated with loss of capillary density (22.7 ± 0.001% in FHH and 9.6 ± 0.014% in control rats, n = 7 and 9, respectively), and greater expression of GFAP positive astrocytes and loss of neurons in affected areas. Hypertensive FHH rats took 2.5 times longer to escape from an eight‐arm water maze as compared to control strains. These results suggest that down regulation of Add3 in astrocytes disrupts the actin cytoskeleton to induce reactive astrogliosis and may affect Aβ clearance, all of which may contribute to neurodegeneration and cognitive deficit associated with aging and hypertension in FHH rats.Support or Funding InformationThis study was supported by grants HL36279 (RJR) and DK104184 (RJR), AG050049 (FF), P20GM104357 (RJR, FF ) from the National Institutes of Health; 16GRNT31200036 (FF) from the American Heart Association. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ang II‐Mediated Regulation of AGT Protein Synthesis in Rat Brainstem Astrocytes is βarrestin1 Dependent
Authors: Negussie Shmuel , Lymperopoulos Anastasios , Clark Michelle Alexa 
Published: June 2021

ObjectiveStudy the role of βarrestins in Angiotensin (Ang) II‐mediated regulation of angiotensinogen (AGT) protein synthesis in astrocytes isolated from the brainstem of spontaneously hypertensive rats (SHRs) or normotensive rats.BackgroundThe βarrestins (βarrestin1 and‐2) terminate G protein signaling by G protein‐coupled receptors (GPCRs) and, at the same time, they initiate their own “second wave” of signaling from GPCRs, independently of G proteins. Ang II, through its cognate GPCR, Angiotensin II type1 receptor (AT1R), signals via both G proteins and βarrestins in various cell types. In our previous study, we reported that SII (an Ang II peptide analog which is a βarrestin‐biased agonist), similarly to Ang II, upregulates AGT synthesis in a time‐dependent manner. Here, we sought to delineate whether Ang II‐mediated regulation of AGT protein synthesis in brainstem‐derived astrocytes is G protein or βarrestin or both G protein and βarrestin dependent.MethodsPrimary cultures of brainstem astrocytes were isolated from the brains of 2–3 days old pups. Double‐stranded siRNAs (Dharmacon) targeting rat βarrestin1 or βarrestin2 were used to knockdown βarrestins. Non‐silencing siRNA (Dharmacon) was used as a control. RNiMAX (Invitrogen) was used as a transfection reagent. Ang II (100 nM) and 10 μM SII (the βarrestin‐biased agonist) were used to study AT1R‐mediated AGT protein expression. Protein analysis was done using Western blotting and mRNA measurement was done using quantitative real‐time PCR. The results in SHR astrocytes were compared to normotensive rat astrocytes.ResultsOur results indicate that βarrestin1 is the major βarrestin isoform in astrocytes isolated from both SHR and normotensive rat brainstem astrocytes. Ang II upregulated AGT protein synthesis in both SHR and normotensive rat astrocytes. This effect was higher in SHR as compared to normotensive rats. siRNA mediated knockdown of βarrestin1 abolished Ang II‐mediated AGT protein synthesis. However, βarrestin2 knockdown did not inhibit Ang II‐mediated AGT protein synthesis.ConclusionIn this study, we showed that βarrestin1 plays a major role in the Ang II‐AT1R‐mediated regulation of AGT protein synthesis in rat brainstem astrocytes in both SHR and normotensive rat models, which could be an important therapeutic target for the treatment of hypertension and other cardiovascular diseases.Support or Funding InformationFunding for this project was provided through a Nova Southeastern University President's Faculty Research and Development Grant (PFRDG)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Toll‐Like Receptor 9 (TLR9) Is Associated with Vascular Dysfunction in 2‐Kydney 1‐Clip (2K1C) Hipertensive Mice.
Authors: Rodrigues Fernanda Luciano , da Costa Rafael Menezes , Silva Carlos Alberto Aguiar , Bomfim Gisele Facholi , Carneiro Fernando Silva , Tostes Rita , Junior Rubens Fazan , Salgado Helio Cesar 
Published: June 2021

IntroductionRenovascular hypertension is the leading cause of secondary hypertension and commonly resistant to antihypertensive therapy. The involvement of toll‐like receptors 9 (TLR9) has already been demonstrated in spontaneously hypertensive rats (SHR) and their activation seems to contribute to the increase in arterial pressure (AP). Therefore, we hypothesized that TLR9 are activated in two‐kidney one‐ clip (2K1C) hypertension, eliciting inflammation and vascular dysfunction, playing a role in the increase of the AP.Methods and ResultsWild type C57BL (WT) and TLR9 knockout (TLR9_KO) mice were anesthetized with isoflurane and were implanted with a silver clip (0.12 mm) around the left renal artery (SHAM operated mice were not implanted with the clip). After 4 weeks, the mice were also anesthetized with isoflurane for direct AP measurement. Afterwards the third‐order mesenteric resistance arteries (MRA) were removed for vascular function evaluation. AP was higher in 2K1C WT mice (133 ± 2 vs 93 ± 4 mmHg in SHAM WT mice), while this increase was partially prevented by the absence of TLR9 (114 ± 5 mmHg in 2K1C TLR9_KO mice). Vascular dysfunction, characterized by increased contractile responses to phenylephrine (Emax: 167 ± 2.2 vs 126 ± 3.5 KCl % in SHAM WT mice) and reduced relaxation to acetylcholine (Emax: 69 ± 1.6 vs 88 ± 1.4 PE % in SHAM WT mice) and sodium nitroprusside (pD2: 7.31 ± 0.07 vs 7.85 ± 0.05 in SHAM WT mice) was found in 2K1C WT mice but not in 2K1C TLR9_KO mice (Emax: 128 ± 2.9 KCl %, 89 ± 2.0 PE % and pD2: 7.78 ± 0.06, when compared to SHAM WT mice, to phenylephrine, acetylcholine and sodium nitroprusside, respectively).ConclusionTLR9 play an important role in vascular dysfunction and AP increase in 2K1C hypertension.Support or Funding InformationFinancial support: FAPESP (Proc. 2015/21976‐1, 2013/20549‐7 and 2016/11988‐5).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulation of Gastrointestinal Smooth Muscle Motility by Losartan in Rats
Authors: Aigbe Flora Ruth , Tobechukwu Mercy Ezinwa , Akindere Olanrewaju Kingsley , Chijioke Micah Chimeremeze , Jaiyesimi Temitope Jonathan , Shonubi Fatima Ayodeji , Adeyemi Oluwafunmilayo Olaide 
Published: June 2021

A drug‐induced gastrointestinal disturbance is an important adverse drug reaction. It has been reported to occasionally present with inflammatory bowel disease‐like manifestations. The mechanism by which this occurs for several drugs is yet to be fully understood. Time and money are sometimes wasted in the diagnosis and treatment (owing to inappropriate strategies) of gastroenterological cases that are simply drug‐induced. Proper assessment of the gastrointestinal actions of drugs would contribute to the knowledge required to significantly address this situation. Losartan is an antihypertensive implicated in diarrhoea, constipation and abdominal cramps, mechanisms of which are yet to be well understood. This preclinical study was carried out to examine the action of losartan on gastrointestinal smooth muscles using rat tissues. Albino rats' caecal and duodenal tissues (4–5 cm) were excised, suspended in tissue baths containing aerated Tyrode physiological solution and connected to Powerlab data acquisition system via isometric force transducers. The direct effect of losartan (0.332–990 μg/ml) and its influence on contractile responses to acetylcholine (1.83×10−5–5.45×10−3 μM), carbachol (1.82×10−4–5.42×10−2 μM) and potassium chloride (2.28×10 – 6.64×10 μM) were determined. Concentration‐dependent contraction of the rats' caecum and duodenum tissues by losartan was observed; with median effective concentration (EC50) values of 8.30 μg/ml and 8.41 μg/ml for both tissues respectively. Losartan significantly (p<0.01) reduced acetylcholine‐induced contraction of the rats' caecum. A significant (p<0.0001) reversal of carbachol‐induced contraction by losartan at 990 μg/ml was observed in the caecum and duodenum tissues. Significant (p<0.0001) reduction in potassium chloride‐induced contraction of the duodenum was also noted, and a similar trend was observed with the caecum. These findings demonstrate that losartan modulates intestinal smooth muscle tone by interfering with one or more of the several pathways involved in gastrointestinal motility. One pathway that seems likely from the observations made in this study is the cholinomimetic pathway. Additional studies to further elucidate the involvement of this pathway and other mechanisms through which losartan mediates its action would enhance understanding of its pharmacology in this regard. The knowledge thus gained would allow for possible optimization of its therapeutic benefit to mankind, as a more rational approach to its use would become possible, due to the availability of additional relevant information.Support or Funding InformationThis work was carried out without external funding.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Snake Venom‐Derived Hemocoagulase Modified with Factor Xa Restores Hemostasis in Heparin‐Treated, Hypocoagulant Mice
Authors: Jackson Madeline , Adili Reheman , Dai Xiangrong , Li Benjamin Xiaoyi , Holinstat Michael 
Published: June 2021

BackgroundExcessive, post‐traumatic bleeding is the leading cause of potentially preventable death in the United States. Nearly one‐third of trauma patients present with coagulopathy upon hospital admission, which significantly increases the incidence of multiple organ failure and death when compared with healthy patients with similar injuries. A hemocoagulase, such as batroxobin, is a thrombin‐like serine protease that is derived from the venom of either the Bothrops Jararaca or Bothops atrox snake and has the ability to convert fibrinogen to fibrin, thus enhancing clot formation and ultimately reducing blood loss in hemorrhagic injuries. Slounase is a modified hemocoagulase combined with Factor Xa (FXa) which may have better hemostatic efficacy in hypocoagulant conditions compared to batroxobin alone. However, the effects of this modified hemocoagulase in the presence of a coagulopathy has not been well characterized in vivo due to lack of reliable hemostasis models.AimTo determine the hemostatic efficacy of slounase compared to both batroxobin and FXa in hypocoagulant murine models using real‐time intravital microscopy.MethodsThe effects of slounase, batroxobin, and Fxa, at doses of both 1 U and 0.1 U, were assessed in vivo by laser ablation of arterioles in the cremaster muscle under real‐time microscopy. Prior to injury, mice were intravenously injected with 25 U heparin to induce hypocoagulant conditions.ResultsIntravenous injection of 25 U heparin inhibited clot formation, significantly increasing bleeding time in untreated, hypocoagulant mice. Injection of slounase, FXa, and Batroxobin did not cause platelet activation in absence of injury. After arteriole injury, FXa did not have a significant effect on clot formation, as FXa alone is inhibited by heparin. Batroxobin‐treated mice had some fibrin formation, however, there was no noticeable change in platelet recruitment. Slounase, at both 0.1 U and 1 U, resulted in a significant enhancement of platelet‐fibrin content, restored clot formation, and significantly shortened bleeding time.ConclusionsThis in vivo study clearly demonstrated that slounase restores hemostasis in hypocoagulant mice by enhancing both platelet activation and fibrin formation, while batroxobin alone only enhances formation of fibrin. Our data strongly suggests that slounase has a better hemostatic efficacy in heparin‐induced hypocoagulant conditions than hemocoagulase alone.Support or Funding InformationThis work was funded by LEE'S PHARM and in part through the National Institutes of Health grants: R01 GM105671 (MH), R01 HL114405 (MH), and R01 MD007880 (MH)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Marine Pharmacology and Pharmaceuticals Pipeline in 2017
Authors: Trieu Jefferson , Newman David J. , Glaser Keith B. , Mayer Alejandro M. 
Published: June 2021

The ocean's organisms continue to deliver a chemical biodiversity that contributes to both the global preclinical and clinical pharmaceutical pipelines. Thus, in November 2017, the clinical marine pharmaceutical pipeline consisted of six marine‐derived drugs approved by the United States Food and Drug Administration (FDA): for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004); for cancer, eribulin mesylate (Halaven®, FDA‐approved 2010), brentuximab vedotin (Adcetris®, FDA‐approved 2011), and trabectedin (Yondelis®, FDA‐approved 2015). Furthermore, there were 23 marine‐derived compounds in clinical trials: 6 marine‐derived compounds in Phase III, 8 compounds in Phase II, and at least 9 compounds in Phase I, many of them auristatin‐containing antibody drug conjugates. Regular updates on the clinical marine pharmaceutical pipeline are posted at http://marinepharmacology.midwestern.edu/clinPipeline.htm. Furthermore, the global preclinical marine pharmacology pipeline continues to generate considerable data on multiple pharmacological classes, as highlighted in a recent review: A. M. S. Mayer, A. D. Rodríguez, O. Taglialatela‐Scafati and N. Fusetani. Marine Pharmacology in 2012–2013: Marine Compounds with Antibacterial, Antidiabetic, Antifungal, Anti‐Inflammatory, Antiprotozoal, Antituberculosis, and Antiviral Activities; Affecting the Immune and Nervous Systems, and other Miscellaneous Mechanisms of Action. Marine Drugs 15 (9): 273, 2017. We thus conclude that in November 2017, both the marine pharmacology preclinical and clinical pharmaceutical pipelines continued to remain very active.Support or Funding InformationSupported by Midwestern University.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐allergic effect of elecampane camphor from Saussurea lappa in ovalbumin‐induced asthma model
Authors: Nam So‐Yeon , Lee Bo‐Kyung , Im Dong‐Soon , Park Soo‐Jin , Kang Saeromi 
Published: June 2021

BackgroundIn Oriental countries, the root of Saussurea lappa is used to treat asthma, rheumatism, and other conditions. Extracts of Saussurea lappa were reported to alleviate house dust mite‐induced atopic‐like dermatitis in Nc/Nga mice, and sesquiterpene lactones were isolated.METHODSTo elucidate whether elecampane camphor, a constuent of Saussurea lappa extract has beneficial effects on allergic asthma, anti‐asthma effects were studied using female Balb/c mice and rat RBL‐2H3 mast cells. Antigen‐induced degranulation was measured in vitro by measuring β‐hexosaminidase activity. A murine ovalbumin‐induced allergic asthma model was used to test the in vivo efficacy of the elecampane camphor.RESULTSAntigen‐induced degranulation was inhibited by the elecampane camphor. Sensitization and challenge of ovalbumin induced allergic asthma responses. Administration of elecampane camphor decreased numbers of inflammatory cells, especially eosinophils, and reduced the expression and secretion of Th2 cytokines, including IL‐4 and IL‐13, in lung tissues and bronchoalveolar lavage fluid. Histologic studies showed administration of elecampane camphor reduced inflammatory signs and mucin production in lungs.CONCLUSIONThe findings provide evidence that elecampane camphor has a potential use as anti‐allergic therapeutics.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.