bluuebunny/crossref_metadata_2025_split huggingface dataset:
Punica Granatum Rind Extract Promotes Mouse Liver Mitochondrial Function and Attenuates Paraquat Toxicity in a Caenorhabditis Elegans Model
Authors: Mowery Colin , Ramanathan Ranjith , Johny Anup 
Published: June 2021

Punica granatum (pomegranate) has drawn the interest of the scientific community due to its rich array of phenolic compounds and antioxidant capacity. Evidence supports the fruit's role in preventing free radical damage in humans. Although various studies have concluded the beneficial effects of pomegranate juice, limited studies have characterized the antioxidant effects of its rind (outer covering/peel; PRE). The goal of our study was to characterize the effects of PRE on oxidative stress. We utilized mouse liver mitochondria and paraquat (PQ) toxicity in a Caenorhabditis elegans (C. elegans) model to determine the beneficial effects of PRE. Mitochondria were isolated from C57 male mice liver tissue (6 weeks of age). Phosphatidylcholine/cholesterol liposomes (Lp) were used to induce oxidative stress. Samples were then incubated at 37□ for 2 hours with or without the addition of PRE at two levels (PRE1 and PRE2 represent 60 and 120 μM Trolox equivalent/mL, respectively). Oxygen consumption rate (OCR; nmol O2 consumed/min per mg mitochondria) was used to express the activity of NADH‐CoQ oxidoreductase, succinate dehydrogenase, and cytochrome c oxidase (complex I, II, and IV of electron transport chain, respectively). Antioxidant level in PRE was determined by DPPH‐, ABTS‐ and Folin‐Ciocalteu assays. The experiment was repeated four times. Pre‐incubation of mitochondria with Lp decreased (p < 0.05) OCR by 50% compared with control mitochondria without Lp. However, incubation of Lp with PRE2 increased (p < 0.05) OCR for complex II and IV. No significant changes (p > 0.05) in OCR were observed for complex I with incubation of Lp and PRE 1 or PRE2. Using C. elegans, an oxidative stress resistance assay was performed to explore PREs effects on paraquat toxicity. Wild‐type adult/L4 hermaphrodites were placed in 96‐well plates. PQ was dissolved to a final concentration of 50 μM and half concentration of PRE1 was added for experimental groups. Resistance to oxidative stress was measured through percentage survival at hour time intervals. After 5 hours, mean survival for PQ alone was 61.91±6.56%; PQ groups supplemented with PRE had a mean survival of 88.29±6.18%. The current study suggests that PRE may play a role in preserving mitochondrial function during oxidative stress and attenuates PQ toxicity.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Specific Profiles Of Circulating Mediators Characterize Older Persons With Physical Frailty And Sarcopenia
Authors: Calvani Riccardo , Picca Anna , Marini Federico , Biancolillo Alessandra , Gervasoni Jacopo , Persichilli Silvia , Primiano Aniello , Landi Francesco , Bernabei Roberto , Marzetti Emanuele 
Published: June 2021

Chronic inflammation, changes in body composition, muscle loss (sarcopenia) and decreasing homeostatic reserve (frailty) are hallmarks of aging. Several circulating inflammatory markers have been associated with these age‐related conditions. However, a gold standard biomarker able to predict functional impairment in older adults is currently unavailable. Muscle is crucial for several metabolic processes, including protein/aminoacid metabolism. Perturbations in protein/aminoacid metabolism may play also a role in the development of physical frailty and sarcopenia (PF&S). The simultaneous analysis of an array of circulating aminoacid/metabolites and their relationship with inflammatory mediators may help gain insights in the pathophysiology of PF&S.To this aim, we analyzed the profile of circulating inflammatory mediators and amino acids in older people with and without PF&S.More than five hundred persons aged 70+ years were screened. Of these, sixty (20 men and 40 women; mean age 76.9±4.8 years) were diagnosed with PF&S. Thirty (14 men and 16 women) non‐sarcopenic, non‐frail persons were enrolled in the control group. A panel of 27 cytokines, chemokines and growth factors was analyzed via a multiplex, magnetic bead‐based immunoassay on a Bio‐Plex® System with Luminex xMap Technology. A panel of 37 serum amino acids and derivatives was assessed by UPLC‐MS. Multi‐block partial least squares discriminant analysis (PLS‐DA) was employed to explore the relationship among inflammatory and amino acid profiles of people with PF&S. Double cross‐validation procedures were used to validate the predictive ability of the PLS‐DA model.The optimal complexity of the PLS‐DA model was found to be three latent variables. The proportion of correct classification was 85.5 ± 4.4 for persons with PF&S and 88.3 ± 3.6 for controls. As for the amino acid profile, people with PF&S showed higher levels of aspartic acid, asparagine, taurine, citrulline, alpha‐aminobutyric acid, methionine, and glutamic acid. Increased levels of interleukin‐8, myeloperoxidase, and platelet derived growth factor‐BB were also found in people with PF&S.The dissection of these patterns may provide novel insights into the role played by inflammatory mediators and protein/amino acid perturbations in the disabling cascade associated with PF&S and possible new targets for interventions.Support or Funding InformationThis study was supported by the “Centro Studi Achille e Linda Lorenzon”, intramural research grants from the Catholic University of the Sacred Heart (D3.2 2013 and D3.2 2015), Innovative Medicines Initiative Joint Undertaking (IMI‐JU 115621), and Fondazione Roma (NCDs Call for Proposal 2013)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Internalization and intracellular trafficking of an antitumor molecule
Authors: Morais Katia , Goldfeder Maurício Barbugiani , Sciani Juliana Mozer , Chudzinski‐Tavassi Ana Marisa 
Published: June 2021

Amblyomin‐X is a proteasome inhibitor that was identified through the transcriptome analysis of the salivary gland from Amblyomma sculptum tick. This recombinant protein triggers apoptosis in different tumor cell lines, and promotes regression of tumor growth and reduction of metastasis. Recent studies have indicated endocytosis mechanism could be associated with pro‐apoptotic activity of this protein. Here, by using confocal microscopy and high content analysis (ImageXpress Micro Confocal High‐Content, Molecular Devices), we found that Amblyomin‐X (Alexa Fluor 488 conjuaged) preferentially co‐localize with Alexa Fluor 555 Cholera Toxin Subunit B, caveolin‐1 and 19S proteasome subunit in human tumor cells (SK‐MEL‐28 and MIA PaCa‐2). Remarkably, we identified Amblyomin‐X co‐immunoprecipitates with caveolin‐1 and Rab‐11. Also, we observed an alteration in Ambyomin‐X trafficking in human tumor cells transfected with GFP‐Rab5 mutants (Rab5:Q79L and Rab5:N133I). Taken all results together, we can suggest that Amblyomin‐X internalization occurs via lipid raft pathway and intracellular trafficking involved of caveosomes and early and recycling endosomes. Then, Amblyomin‐X reaches proteasome system, a possible intracellular destination.Support or Funding InformationSupported by CAPES, Fapesp, BNDES and União Química FarmacêuticaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemical X promotes the osteogenesis and inhibits the adipogenesis through inhibition of lipid metabolic eyzyme
Authors: Jang Hyun‐Jun , Suh Pann‐Ghill 
Published: June 2021

When mesenchymal stem cells differentiate into adipocytes or osteoblasts, the lipid profile changes dynamically and the expression of lipid metabolic enzymes also varies. We found that certain lipid metabolite increased during adipogenesis and decreased during osteogenesis. Consistent with the metabolite change, mRNA and protein of its synthase X also respectively up‐regulated or down‐regulated during differentiation. To examine the roles of the synthase X on differentiation, we examined the effect of the synthase X‐deficiency on adipocyte and osteoblast differentiation by using RNA interference. Knockdown of the synthase X inhibited the adipogenesis and enhanced the osteogenesis. Interestingly, we found that ectopic expression of the synthase X increased PPAR‐γ activity without the expression change of the PPAR‐γ. This increased PPAR‐γ activity was blocked by chemical X, enzyme activity inhibitor of the synthase X, and the product of the synthase X induced the activation of PPAR‐γ. The product of the synthase X also rescued the inhibited adipogenesis by knockdown of the synthase X and it also inhibited the osteogenesis. These effects of the synthase X on fat and bone formation was confirmed in vivo mouse model by using chemical X. Chemical X reduced the body weight and fat mas and accelerated bone repair in a calvarial defect model. Taken together, these results suggest that the synthase X is involved in adipocytes and osteoblasts differentiation through activation of PPAR‐γ by its product and the inhibition of synthase X, by treatment of chemical X, could be the therapeutic options for bone repair.Support or Funding InformationThis work was supported by a grant from the National Research Foundation of Korea (NRF) funded by the Korean government (NRF‐2014M3A9D8034459 and NRF‐2017R1E1A1A01074510) and by Korea Mouse Phenotyping Project (2016M3A9D5A01952411) of the Ministry of Science and ICT through the National Research Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fasciola hepatica Glutathione S‐Transferase (FhGST) conformational studies and its effect on the suppression of toll‐like receptors stimulation
Authors: Valdes Fernandez Bianca N , Aguayo Vasti , Jimenez Caleb Ruiz , Ramos Marcos , Espino Ana 
Published: June 2021

Fasciola hepatica is a parasite well known as a master of immunomodulation because of its ability to suppress the Th1 immune responses and elicit a strong Th2/Treg response. This occurs when the parasite releases excretory‐secretory products (ESPs), which are comprised of a myriad of glycoproteins that are believed to play an important role in the immune response. F. hepatica Glutathione S‐Transferase (FhGST) is one of the ESP components recognized for its anti‐oxidant properties and its capacity to provide part of the parasite's defense by detoxifying the secondary products of lipid peroxidation that are produced due to the immune free‐radical attack on the host or the parasite membrane. Our preliminary studies demonstrate that FhGST exerts a suppressive effect on the activation of TLR4 in macrophages which is evidenced by significant suppression of NF‐kB activation and down‐regulation of TNFα and IL1‐β in response to LPS‐stimuli. We speculated that the integrity of the tertiary structure, as well as the glycans that decorate the protein moiety of FhGST, could be essential for its capacity to suppress the NF‐kB activation. To test our hypothesis, we first subjected FhGST to a thermal denaturing process by boiling it at 90°C for 10 min followed by a treatment with PNGase F. Circular dichroism analysis (CD) comparing the FhGST spectra before and after heat denaturing served to confirm that FhGST was full and irreversibly denatured. Next, denatured or native FhGST were added alone or in the presence of the whole extract of 1 x 108 cells Klebsiella pneumonia (Kp) to a culture of human monocytes (THP1‐CD14) that express most TLRs as well as an NF‐kb inducible secreted embryonic alkaline phosphatase reporter gene. Cells stimulated with an attenuated whole extract of 1 × 108 cells Klebsiella pneumonia (Kp) were used as activation control. Our results demonstrated that native FhGST suppresses NF‐kB activation by 83%, whereas the heat‐denatured protein suppresses NF‐kB activation by 84%. These suppressions suggest that in contrast to expected, the capacity of FhGST to suppress the NF‐κB activation in THP1‐CD14 cells is independent of the integrity of its secondary/tertiary structure and is not associated to the glycans. Docking analysis in which the FhGST‐protein moiety was docked with the CD14 and MD2 co‐receptor structures suggested that FhGST could target these co‐receptors as part of its mechanisms of action.Support or Funding InformationThis study was supported by MBRS‐RISE R25GM061838‐13, 5R25GM061151‐16 and NIH grants G12MD007600 and 2P40OD012217This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Design of SCI‐57: An Ultrastable, Rapid‐acting, Single‐chain Insulin Analog
Authors: Park Madeline 
Published: June 2021

There are approximately 415 million people with Diabetes Mellitus worldwide and there exists a growing need for an ultrastable, heat resistant formulation of rapid‐acting insulin to treat the millions of people who live in developing countries where temperatures routinely exceed 45ºC. The heat resistance and stability of insulin is especially crucial due to the fact that insulin degrades ten times faster for every 10°C increase in temperature above 25°C. Several rapid‐acting insulin analogs have been created, though most do not last longer than 30 days and are reliant on the expensive cold chain delivery system of insulin analogs. This is due to chemical and physical degradation by way of deamidation, transamidation and cross‐beta assembly of linear polymers, or fibrillation. Rapid‐acting analogs must also combat the formation of the inactive hexameric formulation of insulin due to the presence of zinc and an excess of hydrogen ions. A single‐chain insulin analog has been created called SCI‐57 which consists of a glycine rich peptide (6 residues in length) that connects the A and B chains of the insulin analog. Adding a peptide tether will restrict splaying of the A and B chains, thus decreasing the risk of fibrillation even under high temperatures. To ensure that the analog is rapid‐acting and will not form storage hexamers, SCI‐57 has replaced ThrA8 with His to omit an unfavorable beta‐branched amino acid and HisB10 with Asp to create a more favorable relationship with the electrostatic dipole of the B‐chain alpha helix. Replacing HisB10 with Asp also blocks zinc binding, furthermore ensuring that the analog in its active, monomeric state. Both of these actions triple the activity of two‐chain analogs which will also benefit SCI‐57 by allowing it to bind quickly to the insulin receptor. Furthermore, SCI‐57 is much more resistant to fibrillation under high temperatures than previous analogs and maintains insulin receptor affinity as well as efficiency. The Nueva School MSOE Center for Biomolecular Modeling MAPS Team used 3‐D modeling and printing technology to examine structure‐function relationships of SCI‐57.Primary Citation:Hua, Qing‐xin, et al. “Design of an Active Ultrastable Single‐Chain Insulin Analog: SYNTHESIS, STRUCTURE, AND THERAPEUTIC IMPLICATIONS.” The Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 23 May 2008This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Use of Single‐Domain Antibodies from Camelidae for the Inhibition of Clostridium difficile Toxins A and B
Authors: Wall Susan , Akman Alexandra , Chafe Zachary , Chow Alyssa , Deng Mirella , Dumitrascu Flavia , Erisoglu‐Akyildiz Kayra , Hernandez‐Oviedo Isabela , Hu Kevin , Muscant Rebecca , Nguyen Tanya , Niu Catherina , Rulko Theo , Samad Hana , Sullivan Nicholas , Wa Ewan , Wu Yuxin , Xiao Mason 
Published: June 2021

Clostridium difficile is an anaerobic, gram‐positive bacterium and a common gastrointestinal pathogen. Two virulence factors of C. difficile known to mediate disease are Toxin A (TcdA) and Toxin B (TcdB). These toxins are glucosyltransferase‐containing multi‐domain proteins that enter the host's epithelial cells by binding to cell–surface carbohydrate antigens. Once inside the cell, the toxins undergo a pH‐induced conformational change and release their glucosyltransferase domains (GTD), leading to disruption of cell cytoskeleton integrity.Existing treatments for C. difficile such as fecal transplants and antibiotics have not proven highly effective. Passive immunotherapy with inhibitory antibodies recognizing TcdA and TcdB is a promising alternative. Recombinant Camelidae antibodies (VHHs) that neutralize the toxins' function have been identified. TcdA's receptor‐binding domain has seven repetitive elements known to interact with cell‐surface carbohydrates. The VHH antibodies A20.1 and A26.8, which are known to neutralize TcdA, do not block the carbohydrate‐binding site on TcdA directly, suggesting a novel mode of TcdA inhibition.The Ashbury College MSOE Center for BioMolecular Modeling SMART Team used 3‐D modelling and printing technology to examine the structure‐function relationships of a TcdA fragment (TcdA‐A2) simultaneously bound to A20.1 and A26.8. The structure shows that the VHHs target distinct epitopes on the toxin and neutralize TcdA by a novel mechanism independent of the carbohydrate binding sites, providing a rationale for designing highly potent biparatopic TcdA‐neutralizing antibodies.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Yo GABAA GABAA: How the Structure of Human GABAA Receptor Affects the Action of Anesthetics
Authors: Strandberg Stacey , Fleischmann Scott , Bauer E. , Franczak S. , Fricker E. , Griffith‐Topps A. , Jessick M. , Kuehn C. , Lois C. , Sargent C. , Schraufnagel L. , Strandberg S. , Strother C. , Strother J. , Thota L. , Urban S. , Peoples Robert 
Published: June 2021

Propofol, a powerful anesthetic, can be used safely in medical settings, but proves deadly when used recreationally. Propofol binds to the GABAA receptor, which consists of an integral ion channel protein embedded in the membrane of neurons in the brain that is activated by the neurotransmitter molecule gamma‐aminobutyric acid, or GABA. When propofol binds to the GABAA receptor, a conformational change occurs, holding the neurotransmitter GABA in its binding site and keeping the ion channel open. This allows more chloride ions to diffuse into the cell. Resting potential becomes more negative, so even with the diffusion of Na+ ions during nerve stimulation, the threshold cannot be reached and the action potential is not generated. Since the neurons cannot communicate normally, a person given propofol remains unconscious. The lower part of the GABAA receptor, located in the cytoplasm of a neuron, has an unidentified molecular structure. Phe 393 variants in the A and D chains prevent propofol from acting on the GABAA receptor. The DSHA SMART (Students Modeling A Research Topic) Team modeled the similar nicotinic receptor using 3D printing technology to better understand the structure of the GABAA receptor. A greater understanding of the structure of the GABAA receptor and the role of the Phe 393 variants in the action of propofol can lead to the development of more effective anesthetics.Support or Funding InformationThe MSOE Center for BioMolecular Modeling would like to acknowledge and thank the National Institutes of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973) for their support in funding the 2017–2018 SMART Team Team program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evolution of structure‐function relationships in modern estrogen receptors and the reconstructed Ancestral Steroid Receptor 1
Authors: Peterson Alison , Bunday Luke , Bussmann Erika , Chadha Saahil , Ridler Jenna , Romashkova Elena 
Published: June 2021

Ancestral Steroid Receptor 1 (AncSR1) is a reconstructed nuclear receptor protein that gave rise to the steroid hormone receptor (SR) family and which is believed to have predated the divergence of vertebrates from chordates 525 million years ago. AncSR1 is a transcription factor that regulates the expression of proteins implicated in a broad array of biological functions, including reproduction and immunity. AncSR1's ligand binding domain has high specificity for steroid ligands with aromatized A rings and can recognize such molecules in nanomolar quantities. The evolution of specificity by the ligand binding domain led to the divergence of other subclasses of the steroid receptor family. However, AncSR1 (in addition to all proteins within the SR family) follows the minimal specificity model, evolving greater specificity only when it becomes advantageous to discriminate between the various ligands to which it is exposed. The Minnetonka MSOE CBM SMART team used 3D modeling and printing technology to examine structure‐function relationships and AncSR1's place in the evolution of SR proteins. AncSR1 was overlaid with modern estrogen receptors to explore structural evolution in the family and compared with previous findings to determine relation of structural evolution to function. Active study of structural evolution in AncSR1 and the broader SR family is leading to the development of improved pharmaceutical ligands.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In with the Good and out with the Bad – The Role of SR‐B1 in Lowering Blood Cholesterol Levels
Authors: Tiffany Karen , Arnholt A. , Arzumanyan D. , Arzumanyan L. , Barber M. , Copes E. , Crow T. , Esser E. , Kuborn A. , Reinhardt K. , Tiffany B. , Woods A. , Sahoo Daisy 
Published: June 2021

According to the CDC, 1 in 3 adults in the US has high cholesterol, which is linked to cardiovascular disease. Cholesterol, a hydrophobic molecule, is transported in the blood by binding a lipoprotein. Scavenger receptor class B type 1 (SR‐B1), a transmembrane protein that binds high density lipoprotein (HDL) carrying cholesterol, helps transfer cholesterol from HDL into liver cells where it can be excreted in bile. This is important for lowering blood cholesterol levels and reducing the risk of heart disease and stroke. SR‐B1 belongs to a larger family of scavenger receptor proteins that includes lysosome membrane protein 2 (LIMP‐2), and SR‐B1 and LIMP‐2 share structural homology. The structure of the C‐terminal transmembrane domain of SR‐B1, determined through NMR, is composed of three helices with a leucine zipper motif. Mutagenesis studies implicate this leucine zipper motif in dimerization of SR‐B1 and proper receptor function. A crystal structure of LIMP‐2 revealed the existence of a large, primarily hydrophobic cavity that runs the entire length of the protein that suggests a role in the selective transfer of cholesterol into the liver cell. Using 3D printing technology, the Cedarburg SMART (Students Modeling A Research Topic) Team utilized the known structure of LIMP‐2 to investigate structure‐function relationships in SR‐B1. Determining important HDL/SR‐B1 interactions will increase knowledge of how SR‐B1 functions in transferring cholesterol from plasma HDL to the liver for excretion and can lead to the development of medical treatments to increase cholesterol excretion by the liver and prevent heart disease and stroke.Support or Funding InformationThe MSOE Center for BioMolecular Modeling would like to acknowledge and thank the National Institutes of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973) for their support in funding the 2017–2018 SMART Team Team program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.