bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Hepatocytes Uptake Clearance of Organic Anion Transporting Polypeptide (OATPs) Substrates in Human and Cynomolgus Cryopreserved Hepatocytes
Authors: Liang Xiaomin , Deforest Natalie , Park Yeojin , Hao Jia , Zhao Xiaofeng , Lai Yurong 
Published: April 2020

Transporter‐mediated clearance plays significant roles in drug bioavailability and elimination, and evaluation of potential transporter‐mediated drug‐drug interactions (DDI) is recommended by regulatory agencies during drug development. Currently, the direct extrapolation from in vitro to in vivo is a common approach for human clearance prediction for the compounds that are metabolized by hepatic cytochrome P450 enzymes; however, challenge remains when using the similar approaches in prediction of transporter‐mediated clearance. Recent publications attempted to establish the in‐vitro‐to‐ in vivo scaling based on hepatocyte uptake studies and in vivo pharmacokinetic (PK) data from the preclinical animals. However, the differences in homology of transporter proteins and the associated substrate specificity in transporters between human and preclinical animals complicate the prediction using the correcting factors obtained from the preclinical animals' in‐vitro‐to‐ in vivo scaling. In this study, we obtained the in vitro hepatic uptake clearance in cynomolgus monkey and human for 14 literature compounds that were known as OATPs substrates and had clinical DDI reported. Hepatic uptake was conducted using the oil‐spin method. For each selected substrate, the uptake assay was conducted in the time course from 0.25 minute to 15 minutes in KHB buffer with 10% serum. The uptake clearance was estimated from the linear uptake phase. For each selected substrate, the uptake clearance was characterized in three lots (including single donor and pooled donors lots) of human and monkey cryopreserved hepatocytes. In addition, we tested different lots of human (7 lots) and monkey (10 lots) cryopreserved hepatocytes using rosuvastatin as a control probe to examine the variability among lots and donors. In summary, rosuvastatin uptake clearance in cynomolgus monkey hepatocytes was about 2.5 folds higher on average compared to that in human hepatocytes. Generally, for other OATPs substrates studied, hepatic uptake clearance in cynomolgus monkey appeared to be higher than that in human. Furthermore, variability of uptake clearance was also observed among lots and donors. The results obtained in the current investigations are useful for human prediction through understanding the source of variations and establishing the correlations of in vitro‐in vivo scaling.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular characterization of eel Lutropin/Chorionic Gonadotropin Receptor mutants
Authors: Byambaragchaa Munkhzaya , Min Kwan‐Sik 
Published: April 2020

Whereas the ligand mutations that have been found to represent usually loss‐of‐function mutations, the G protein coupled receptor mutations can be both of the gain‐of function and loss‐of function type. There are a lot of number naturally occurring mutations of the lutropin/chorionic gonadotropin receptor (LH/CGR) gene associated with reproductive disorders. Although the hormone binding and activation properties of these mutants are generally well characterized in hLHR, little is known about their regulation and signal transduction if it occurs in eel LH/CGR. With this in mind we examined the effects of the several constitutive activation and inactivation mutations which are occurred in highly conserved residues of LH/CGR such as: activation mutations (M410T at TM2; L469R at TM3; D590Y at TM6) and inactivation mutations (D383N at TM2; Y546F at TM5) in eelLH/CGR consisting of 705 amino acid.To assess the functional effects of 5 receptor mutations directly, wild‐type (WT) and mutant receptors were transiently expressed in CHO‐K1 cells. We evaluated the basal and cAMP stimulation by recombinant eel Luteinizing hormone (rec‐eelLH). The constitutive activation mutants all had elevated levels of basal cAMP. But inactivation mutants were remarkably decreased cAMP production in response to rec‐eelLH. Two of activation mutations (L469R and D590Y) apparently activated eelLH/CGR with the 19.8 and 8 fold elevated basal cAMP level. However M410T mutation showed only 4 fold elevated basal cAMP level, it could be possible to activate eelLH/CGR in a weak manner. Maximal responses of activation mutants were 7.2–46.7% lower than that of WT. The EC50 in two inactivation mutations significantly retarded the cAMP accumulation to 42.3 ng/ml and 1181 ng/ml, respectively.Support or Funding InformationThis work was financially supported by the Korean Research Foundation Program (2018R1A2B6007794)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The myeloid lineage‐determining transcription factor PU.1 induces enhancer‐promoter looping that promotes IL‐1β enhancer and messenger RNA production in non‐myeloid melanoma cells
Authors: Kim Sung Ouk , Cho William , Ha Soon‐Duck 
Published: April 2020

The DNA‐binding protein purine rich (PU.1) is a lineage determining transcription factor required for development of myeloid cells such as monocytes/macrophages. PU.1 is regarded as a pioneering transcription factor for macrophage differentiation due to its ability to bind “closed” genomic sites and initiate/maintain “open” chromatin state in macrophage‐restricted response genes. To date, however, precise mechanism of PU.1 in remodeling chromatin is yet to be elucidated. The non‐myeloid melanoma B16.BL6 cells response to the bacterial cell wall component lipopolysaccharide (LPS) and activate the transcription factor NF‐κB, but do not produce cytokines. Ectopic expression of PU.1 rendered these cells express the cytokine interleukin 1‐β (IL1b) in response to LPS. PU.1 induced chromatin remodeling that caused interaction between the IL1b promoter and enhancer (located ~10 kb upstream of the IL1b transcription start site) in non‐activated cells; upon activation, PU.1 recruited NF‐κB to these genomic regions. The N‐terminal domain (1–30) of PU.1 was important for both promoter‐enhancer looping and IL1b eRNA/mRNA production; whereas, the acidic and glutamine rich domains (33–100) were required for eRNA/mRNA production but dispensable for the DNA looping. eRNA production and histone acetylation were required for NF‐κB recruitment and IL1b mRNA expression, but not for the DNA looping. Altogether, this study indicates that PU.1 induces enhancer‐promoter looping that sequentially allows activation‐dependent histone acetylation, recruitment of NF‐κB and production of enhancer RNAs, all of which are required for an optimal IL1b production.Support or Funding InformationThe Natural Sciences and Engineering Research Council of Canada ‐ Discovery Grant (RGPIN‐2018‐05514)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis and the identification of DNA binding sites for the cI repressor of Bacteriophage ɛ34
Authors: McGee Lauren , Gildea Logan , Goodson Richardria , Villafane Robert , Jackson Doba , Brooks Ryan 
Published: April 2020

Gene 46 of ɛ34 encodes a protein that is similar to other phage repressors. It has a C‐terminal protease domain which is about 80% identical to the repressors of phage λ, but its DNA binding N‐terminal portion is only about 18% identical to its closest known relatives in phages λ, and Lex A. The N‐terminal DNA‐binding domain is homologous to the HTH_XRE family of DNA‐binding domains. In another study we have generated homology models of this proteins and its interaction with DNA [1].The main goal of this project was to initiate a structure‐function study on the cI repressor protein of phage ɛ34. The cI repressor of ɛ34 protein with a 6X polyhistidine tag was first isolated and purified to homogeneity. We have identified regions for cI binding by using bioinformatics and gel‐mobility shift assay using PCR fragments encompassing the intergenic region of the cI and cro gene. Our binding sites compare well with the known intergenic regions of other phages such as λ, Φ11, 434. This project was done as a collaborative venture between two laboratories, Huntingdon College (Dr. Doba Jackson) and Alabama State University (laboratory of Dr. Robert Villafane). This project is in line with Dr. Robert Villafane's research into the Genomic analysis of bacteriophage ɛ34 of Salmonella enteric serovar Anatum.Support or Funding InformationSupport Support for this work was provided by Alabama State University and Huntingdon College.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Novel Rigid Glycomimetics to Inhibit Influenza Infection
Authors: Woods Robert J. , Ji Ye , Hendel Jenifer , Smith Hannah , Glushka John , Wu Nicholas , Wilson Ian A. , Tompkins S. Mark , Murphy Paul 
Published: April 2020

Small‐molecule anti‐influenza agents, such as zanamivir and oseltamivir are neuraminidase (NA) inhibitors limit the release of progeny virions from infected cells through competitive binding to the viral NA. However, these drugs do not block infection. Vaccination is the most common approach for preventing influenza infection, but vaccine creation takes approximately 6 months and requires prior knowledge of the circulating virus strains. The impact of this limitation was evident in the 2009 swine flu pandemic, against which the seasonal vaccine was completely ineffective. The urgent need to develop complementary strategies to vaccination or NA inhibition is emphasized by the fact that between 2006 and 2009 seasonal flu developed almost complete resistance to oseltamivir, resulting from a single spontaneous mutation in the viral NA.As a potential alternative to current flu antivirals, we are developing glycomimetics that block the interaction between the viral hemagglutinin (HA) and the host glycans that serve as flu‐receptors. This interaction is essential for viral adhesion, and the universal receptor for flu A [15] and B [16] viruses is Sia‐Gal (sialic acid, Sia, a.k.a. neuraminic acid, linked to galactose, Gal). HAs from either human (H1N1) or avian‐infective (N5N1) flu strains bind to di‐ or trisaccharides terminating in Sia‐Gal regardless of whether the Sia‐Gal linkage is α2,6 or α2,3. This observation suggested that a small Sia‐Gal glycomimetic might be developed that could inhibit a broad range of flu A strains, including flu B, and was critical in the design of our glycomimetic framework (FB127).Here were report that FB127 shows dose‐dependent inhibition of the infection of MDCK cells by a range of flu viruses from group 1 (H1N1, H5N1), group 2 (H3N2), and flu B, and that the influenza neutralizing ability of FB127 is within the same range as reported for glycodendrimers. Because FB127 is a synthetic small molecule, it can be rationally modified, and manufactured with a high degree of quality control and reproducibility. In this regard, FB127 is amenable to a number of formulations, including aerosol, oral, and intravenous routes that provide great flexibility in drug delivery.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dysfunction of a microglia subtype produces sex‐linked anxiety
Authors: Traenkner Dimitri 
Published: April 2020

Microglia are immune cells of the central nervous system. Different types of microglia exist, yet we incompletely understand to what extend subtypes serve different functions. To test this, we used a selective genetic ablation strategy in mice. Intriguingly, loss of one microglia subset causes severe anxiety exclusively in females. Moreover, this behavior develops at the time of sexual maturity, suggesting an important role of female sex hormones in this anxiety model. Our findings suggest a mechanistic link between sex and genetics, two major risk factors for anxiety disorders in people.Support or Funding InformationNational Institute of Health grant R01MH093595This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Intralipid Infusion in Pregnant Rats Induces Plasma Angiogenic Imbalance, Inflammation, and Intrauterine Growth Restriction
Authors: Duncan Jeremy W. , Bergeron Matthew , Bradshaw Jessica L. , Spradley Frank T. , Granger Joey P. 
Published: April 2020

Preeclampsia is a pregnancy specific disorder characterized by maternal hypertension and proteinuria and is associated with low birth weight, vascular dysfunction, and an increase in inflammatory and anti‐angiogenic factors. Recent studies have shown that obese women are 4 times more likely to develop preeclampsia. In addition, preeclamptic patients have higher levels of circulating free fatty acids (FFA) and other lipids compared to normal pregnant women. While a high fat diet can lead to reduced uteroplacental perfusion, vascular and placental dysfunction, the mechanism whereby lipids contribute to these outcomes in preeclampsia is unknown. To test the hypothesis that lipid administration induces placental and vascular dysfunction, pregnant rats were infused intravenously with 20% Intralipids (50–100 μL/kg/min) over a 2 h period on gestational days (GD) 11–18 or ‐19. On GD 19, mean arterial pressure (MAP), pup weight, placental weight, blood and tissues were collected. Third order mesentery arteries and aortas were used for vascular reactivity studies, and placental villi were cultured for 48 h to measure the production of angiogenic factors ex vivo. Infusion of intralipids resulted in no change in MAP, vascular reactivity, or placental villous angiogenic factors. However, infusion of intralipids in pregnant rats caused a significant increase in placental weight (0.66 ± 0.01 g vs 0.61 ± 0.01 g; p<0.01) and fetal absorption (13.8 ± 5.9% vs 0.85 ± 0.85%; p<0.05), and a decrease in pup weight (2.37 ± 0.02 g vs 2.57 ± 0.02 g; p<0.01) compared to control rats. Furthermore, plasma tumor necrosis factor‐alpha (TNF‐α; 24.82 ± 6.21 pg/mL vs 12.24 ± 1.52 pg/mL; p<0.05) and soluble fms‐like tyrosine kinase (sFLT‐1; 20.08 ± 5.76 pg/mL vs 4.42 ± 2.04 pg/mL; p<0.05) were increased, while plasma placental growth factor (PlGF; 28.57 ± 5.65 pg/mL vs 51.19 ± 8.82 pg/mL; p<0.05) was decreased in intralipid‐infused pregnant rats compared to control rats. These data indicate that increasing lipid intake during pregnancy induces intrauterine growth restriction coupled with a pro‐inflammatory and anti‐angiogenic state. Further studies to determine the factors associated with intralipid‐induced fetal outcomes are warranted and could provide a better understanding of the role of elevated lipids/FFA as a risk factor for preeclampsia.Support or Funding InformationP01HL051971, P20GM104357, and T32HL105324.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effects of Non‐Small Cell Lung Carcinoma Conditioned Media on Skeletal Muscle Cell Proliferation and Metabolism
Authors: Stott Nicole L , Beach Cassandra , Marino Joseph S 
Published: April 2020

Lung cancer is the second most common cancer and one of the largest causes of cancer‐related mortality worldwide. 80–85% of all new lung cancer diagnoses are Non‐Small Cell Lung Carcinoma (NSCLC), suggesting a great need to continue elucidating the metabolic mechanisms surrounding this prevalent disease. Due to the oncogenic environment surrounding NSCLC growth, factors released into the environment could have a direct impact on other tissues. In particular, skeletal muscle, a leading insulin‐sensitive and metabolically active tissue, is subject to wasting in many cancer patients. The purpose of this study was to determine how NSCLC conditioned media affects C2C12 skeletal muscle cells proliferation and metabolism. C2C12 mouse skeletal muscle cells were grown for 24 hours and serum‐starved overnight with serum‐free media to induce quiescence. Cells were treated with equal volumes of either fresh growth media or conditioned media from proliferating NSCLC cells. C2C12 myoblast proliferation was assessed every 24 hours for 3 days via an MTT assay using 3‐(4,5‐Dimethyltiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide. Proliferating C2C12 myoblasts treated with fresh growth media or conditioned media from NSCLC cells showed significant proliferation (p=0.0016) over 72 hours. Treatment with NSCLC conditioned media did not induce any significant increases in myoblast proliferation when compared to control C2C12 cells. In conclusion, these data suggest that C2C12 myoblasts did not respond adversely to NSCLC conditioned media. Experiments are ongoing to investigate alterations in metabolic and myogenic signaling pathways, as well as extended time courses.Support or Funding InformationThis work was supported by the Targeted Research Internal Seed Program, JSM.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Efficiency vs. Competency Development: What Interprofessional Health Professions Students Learn From, With, and About Each Other During Anatomy Dissection
Authors: Nelsen Sylvia , Tucker Constance 
Published: April 2020

The gold standard for anatomy instruction has traditionally involved a combination of lecture‐based learning and hands‐on discovery through cadaveric dissection. However, a full dissection is time‐intensive, and recent curricular changes are challenging educators to deliver content via more time‐efficient modalities. As a result, laboratory hours are often a place where reductions in instructional time are made. This tension between time and meaningful activity is heightened as curricular innovations such as simulation, 3D visualization, and prosection are shown to reduced teaching time and increase the student: cadaver ratios. However, the dissection experience is not only important for helping students understand the structure and function of the human body. It is also instrumental in helping students develop skills and attitudes related to professionalism such as teamwork, communication, and respect for each other and their future patients, learning opportunities that are often referred to as the “hidden” curriculum (Escobar‐Poni & Poni, 2006; Warner & Rizzolo, 2006). More recently, the ability of healthcare providers to work effectively in interprofessional teams has been added to this list of skills, and dissection experiences have been designed to explicitly incorporate interprofessional experiences into anatomy lab curricula (e.g., Fernandes et al, 2015). To date, the impact of an interprofessional learning environment within the anatomy lab on learners' perceptions of teamwork has not been thoroughly examined. To explore the impact of the interprofessional learning experience, students were asked to write a short essay reflecting on what they learned from their teammates and the donors, and how that will affect the way they approach clinical care and working in professional teams. Constant comparative analysis was used to compare themes from essays written by dental students working in interprofessional teams comprised of dental, physician assistant, and radiation therapy students to those written by dental students working in teams segregated by profession. Results of the study highlight the unique contributions of an interprofessional dissection environment on the learning experience of health professions students.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mediation of Vesicular Fusion by SNARE Proteins during Macroautophagy
Authors: Shannon Daniel , Glass Jen , Bartlett Matt , Bissmeyer Teddy , Bradley Mariah , Buehler Sam , Campman Scotty , Glass Jake , Gutzwiller Leah , Hoffman Trevor , Kellogg Nick , Mahan Erin , Peters Kyle , Poetker Garrett , Schaller Stephanie , Diao JaiJie 
Published: April 2020

The Moeller SMART Team in conjunction with Dr. JiaJie Diao at the University of Cincinnati Medical College and MSOE Center for Biomolecular Modeling used 3‐D modeling and printing technology to study the role of Soluble NSF Attachment Protein Receptor (SNARE) proteins in macroautophagy, a process that recycles damaged organelles, proteins, and microbes in the cell. It begins with the formation and maturation of an autophagosome. The process ends with the fusion of a fully matured autophagosome to a lysosome membrane, which is mediated by SNARE proteins. Vesicular fusion is initiated by the formation of a tetrameric alpha‐helix complex that is formed through the binding of three helical SNARE proteins. One of these SNARE proteins, VAMP8, is located on lysosomes and the other two, syntaxin17 and SNAP‐29, are located on the membrane of autophagosomes. The helices of these proteins coil tightly in the alpha‐helix complex which provides the energy for fusion. To reset the conformation of SNARE proteins, N‐Ethylmaleimide‐Sensitive Factor (NSF) proteins use ATP and water to recycle SNAREs. Alpha‐SNAP, an ATPase, which helps to facilitate this process, returns SNARE to its original high energy state. If fusion is unable to occur in autophagy, the cell accumulates autophagosomes, causing lysosome storage diseases such as Parkinson's and Alzheimer's disease. By 3‐D printing the coiled alpha‐helix complex, our SMART team intends to describe this crucial element of membrane fusion and the disorders associated with macroautophagy.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.