bluuebunny/crossref_metadata_2025 huggingface dataset:
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 Information

This 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 Information

Supported by CAPES, Fapesp, BNDES and União Química Farmacêutica

This 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 Information

This 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 Information

This study was supported by MBRS‐RISE R25GM061838‐13, 5R25GM061151‐16 and NIH grants G12MD007600 and 2P40OD012217

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 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 2008

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 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 Information

The 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 Information

The 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.
Understanding the structure and function of cone snail insulin as a substitute for human insulin
Authors: Lane Jim , Firminger Makayla , Fraser Emily , Geiger Mya , Morris Sydney , Ryan Marin
Published: June 2021

It is estimated that more than 415 million people currently have diabetes. Diabetes is an inability to properly regulate and/or produce insulin. Insulin dependent diabetics need to inject themselves with insulin in order to eat, drink, and function properly. Current insulin activation is around fifteen minutes but can take up to 90 minutes to fully activate. Cone snail insulin (CSI) appears to have the ability to fully activate within minutes. Rapid activation can prevent the body from spending long periods of time in a hyperglycemic state (too much sugar in the bloodstream). Long periods of time in a hyperglycemic state can cause damage to blood vessels and nerves, which in turn, leads to damage to the entire body. The goal of this research process was to discover why and how CSI binds to the receptors and how to mimic this behavior to decrease the activation time of insulin in the human body. The exploration phase of this research inquired how CSI would potentially bind with human insulin receptors and determine if it would be possible to mimic this to use in diabetic patients to decrease the activation time. During the modeling phase of this project, JMOL was used to compare the molecular structures and functions of CSI and human insulin in order to establish if CSI would be interchangeable with human insulin. Cone snail venom contains insulin which causes a hypoglycemic sedation, paralyzing its victim on contact. While researching this process, it was discovered that the B‐chain on the CSI protein is missing the amino acid residues from B23–B30. Human insulin, when stored in the pancreas, aggregates into dimers. These dimers group together to form hexamers. Injected insulin is also in dimer form, but it needs to disaggregate in order to become effective. One human insulin molecule binds to the other using hydrogen bonds on the amino acid residues from B23–B30. Because CSI is missing this portion, it can enter the bloodstream without aggregating. In order to mimic the function of the CSI in humans, human insulin needs to be modified to eliminate the aggregation of insulin cells in the human body. To do this the Mahtomedi High School team, Mahtomedase, MSOE Center for BioMolecular Modeling MAPS Team used 3D modeling and printing technology to examine structure‐function relationships between CSI and human insulin. Current insulin activation time can take, at minimum, fifteen minutes. CSI has the potential to increase reaction time by upwards of 200 percent by more readily binding to the human insulin receptor.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Insulin vs Genetically Engineered Fast Acting Insulins
Authors: Mukhtar Farzan , Veljkovic Petar , Melnyk Max , Smetanin Yakov , Gong David , Lawal Dayo , Drohomyrecky Michael , Mingolelli Gabrielle
Published: June 2021

Insulin is a protein produced in the pancreas. The primary function of insulin is to regulate the levels of glucose in the body from food that is consumed. When food is consumed, beta cells in the pancreas, the body is signalled to release insulin. The insulin that is released binds to insulin receptors, which then cause glucose transporters to be inserted in the membranes of different cells. The glucose transporters allow the cells to take in glucose from the bloodstream, and be used for energy for the body, or be stored for later use as glycogen.

Individuals with diabetes are not able to produce sufficient insulin in the pancreas to keep blood glucose levels low, leading to health problems. One treatment for diabetes is to use insulin medication to control blood glucose levels. Due to the need for insulin medication which quickly takes action and lowers the blood glucose levels of diabetes patients, fast‐acting insulin was developed as a medication option. The structural differences between fast‐acting insulin medication, and insulin that is produced naturally in the pancreas are what cause the two proteins to function differently.

Insulin lispro, which was the first genetically engineered rapid acting insulin, has a slight difference in the B chain than human insulin, where the proline at position 28 is switched with lysine at position 29, resulting in a lower capacity of self association in a solution, which results in a faster absorption, higher peak serum levels, and a shorter action duration than human insulin. Insulin aspart, another form of fast acting insulin, is structurally different from human insulin, in that the proline at position 29 is switched with a charged aspartic acid resulting in it being absorbed twice as fast as human insulin. Insulin glulisine, the most recently created fast acting insulin, is different in structure from human insulin because the asparagine at position 3 is replaced with lysine, and the lysine at position 29 is replaced with glutamic acid, which results in a reduction in hexamer formation and an enhanced absorption from subcutaneous depots. These differences in structure between the three different types of fast acting insulins and human insulin will be directly pointed out with the models of the different insulin types which will be created.

Support or Funding Information

Funded generously by Chicago Public Schools, the Cless Family Foundation, Friends of LIncoln Park High School, and the Center of Biomolecular Modeling at Milwaukee School of Engineering.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Insulin, You're Confusing the Systems
Authors: Bartolomei Barrett A , Mergens Steven Ryland , Love Ryan Ann , Shipley Ashley Willow , Martinez Sandra Ann
Published: June 2021

Insulin is a protein (hormone) that changes conformation to engage its receptor. In 2015 it affected an estimated 1.6 million deaths due to high blood glucose, from diabetes. Inability for insulin to bind to its receptor ultimately leads to the overabundance of glucose. This overabundance then leads to mitochondrial stress in pancreatic beta cells causing unwanted beta cell apoptosis and dysfunction. One component of the insulin change is the decrease in insulin sensitivity in the bloodstream due to glucose overexposure. Insulin becomes resistant to hormonal stimulation suggesting a change in the C‐terminal B‐chain segment. This conformational change insulin undergoes exposes the side chains IleA2, ValA3, ValB12, PheB24, and PheB25 to allow insulin to bind to its receptor. A further understanding of how the native folding is preserved and the binding sites that are blocked are opened, can open opportunities for additional clinical mutations in the insulin family and provide a basis for new research and therapeutic drugs, possibly leading to a more regulated interaction between insulin and glucose.

Support or Funding Information

PDB: 4OGA Primary Citation: Protective hinge in insulin opens to enable its receptor engagement. Menting, J.G., Yang, Y., Chan, S.J., Phillips, N.B., Smith, B.J., Whittaker, J., Wickramasinghe, N.P., Whittaker, L.J., Pandyarajan, V., Wan, Z.L., Yadav, S.P., Carroll, J.M., Strokes, N., Roberts, C.T., Ismail‐Beigi, F., Milewski, W., Steiner, D.F., Chauhan, V.S., Ward, C.W., Weiss, M.A., Lawrence, M.C. (2014) Proc.Natl.Acad.Sci.USA 111: E3395–E3404 The El Capitan High School CBM MAPS Team used 3‐D modeling and printing technology to examine structure‐function relationships of insulin.

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 Analysis and Phenotype of the PTF1‐J/Ptf1a P191T Mutant Heterotrimeric Complex
Authors: Coats Ward , Ali Nor , Ali Arham , Davies Latham , Sterling Jewel , De Leon Luis , Sandoval Ernesto , Pacheco Ariadne , Montes Kristen , Guerrero Ingrid , Jordan Emily , Mulley Alexander
Published: June 2021

The Hillcrest High School Biomedical Research Program in Dallas, Texas, studies the PTF1‐J and PTF1‐JL heterotrimeric complexes. The complexes are composed of a common E‐box binding protein, Ptf1a and RbpJ or RbpJL. These complexes are transcriptional master regulators of pancreas development. PTF1‐J is required for early pancreas development. During the secondary transition of pancreas development RbpJ is replaced by RpbJL. The PTF1‐JL complex is required for the mature acinar cell phenotype. In the current study we have performed structural modeling using Pymol and Jmol of the PTF1‐J and PTF1‐J/Ptf1a P191T complexes. Children born without a functional Ptf1a protein lack a pancreas, are severely diabetic and require immediate medical intervention by administering insulin to maintain blood glucose homeostasis and or enzyme therapy to provide digestive enzyme activity in the small intestine for nutrient uptake into the bloodstream. In the current study we have performed structural modeling of the PTF1‐JL/Ptf1a P191T complex and used 3‐D printing technology to generate a model that may help explain the loss of function resulting in pancreatic aplasia in children born with this mutation.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Augmented Reality as an Educational Tool for Teaching Macromolecular Structure and Function
Authors: Sung Rou‐Jia , Wilson Andrew , Crowl Logan , Nardi Joseph , St. Clair Katie
Published: June 2021

Relating macromolecular “structure to function” is considered a foundational skill in biochemistry and molecular biology course curriculums. The molecular visualization skills necessary to interpret information in 3D models of biological macromolecules, however, are often lacking. Moreover, current technologies aimed at spanning this skills gap often require varying degrees of prior knowledge, training, and/or hardware requirements that can make implementation difficult. To address this we have developed a simple classroom activity using augmented reality technology that allows students to quickly and easily interact with high‐resolution images of a 3D macromolecule. The activity was implemented in a junior/senior level biochemistry course as part of a class discussion on the structure and function of the potassium channel. The 3D images were integrated as an iPad application in which a 3D virtual image of the molecule would appear superimposed on the real world on the iPad screen when the iPad camera was directed towards a printed QR code. Students were able to rotate, translate, and zoom in on the molecule by physically moving either the iPad or the QR code sheet. The activity was also accompanied with worksheet questions aimed at developing an interpretation of the 3D structure. Student attitudes were assessed using a pre/post survey analysis. Results indicate increases in student confidence towards visualizing the 3D structures of biological macromolecules, as well as an increase in positive student perceptions of the helpfulness of looking at 3D structures for learning protein biochemistry.

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 Survey Course for STEM Students to Understand Diversity and Inclusion Through Readings, Discussion and Reflection.
Authors: Benore Marilee Ann , Smith Sheila R
Published: June 2021

Scientific discovery, innovation and success are enhanced when diverse groups participate.

Promoting diversity and inclusion requires understanding both obstacles and best practices. One way to achieve success is to ensure that students in STEM fields recognize bias, understand why a diverse workforce is critical, and know how to support diversity and inclusion.

This hybrid class is an overview of gender in science in engineering. Weekly lectures and in‐class discussion coupled with online discussion with peers are critical components of student learning. The topics include STEM statistics, best practices, research, implicit bias, a brief history of women in STEM, feminist theory, and the science of sex and gender. An emphasis that opinions and statements must be supported by factual references. Primary literature, videos, guest lecturers, interviews, books and white papers are used as sources. The final student project is a proposal to create change via outreach, education, policy or research.

Support or Funding Information

UM Dearborn

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Conversion of Invasive Species, Rhamnus carthartica, into Combustible Biofuel: A Novel Approach to Conservation
Authors: McCollough Shannon , Eickhoff Isaac , Soneral Paula
Published: June 2021

Common buckthorn, Rhamnus carthartica, is an invasive species that outcompetes native species throughout North America. Introduced from Europe for horticultural purposes, this decorative shrub is fast‐growing, shade tolerant, and depletes soil nutrients. Given its exquisite fitness and negative consequences to biodiversity, an approach to mitigate the detrimental effects of buckthorn is essential. Can buckthorn biomass be turned into energy? The goal of this project was to evaluate the efficacy and feasibility of transforming buckthorn into combustible ethanol as a long‐term conservation strategy. We asked two central questions in the context of a course‐based undergraduate research experience: 1) Which tissue ‐ leaves, stems, or berries ‐ is the optimal source of biomass? 2) Does pre‐freezing enhance ethanol yield? Tissue harvested from 20 locations in the upper‐midwest were subjected to cellulase pretreatment followed by fermentation. Crude samples were then filtered and subjected to simple and fractional distillation to 95% purity. The ethanol from all sources was combustible. Although berries contained the highest amount of pre‐cellulase glucose, all biomass produced comparable amounts of final ethanol yields (ANOVA p=.93). Freeze‐thaw pretreatment of these tissues produced significantly lower ethanol yields (T‐test p=.00063). Based on this data, we propose a scalable, sustainable, and economically viable buckthorn‐to‐biofuel conversion program that can be implemented in a variety of community contexts.

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 relationship between student's learning motivation and life style in a Japanese medical school
Authors: Sato Maki , Miyata Yasushi , Waseda Katsuhisa , Aoki Ruri , Kwahara Chikako , Ban Nobutaro
Published: June 2021

Motivation research has studied in educational psychology in past decade (Pintrich, 1993; 2000) in primary, secondary and college education (Vansteenkiste et al., 2005). The motivated strategies for learning questionnaire (MSLQ) is a self‐report instrument designed to assess college students' motivational orientations (Pintrichet al., 1993). MSLQ is based on a general cognitive view of motivation. The studies of learning motivation in medical students were limited.

In this study, we examined the relation between learning motivation and lifestyle such as food intake and sleep in medical students. 421 medical students participated in the present study. The questionnaire was all anonymous. The survey included that the learning motivation, daily food intake and sleep pattern. We compared those items and learning motivation.

In the results, the frequency of breakfast and consumption of vegetable and milk production were correlated with learning motivation, though the sleep duration was not significantly correlated with learning motivation.

In conclusion, the balanced food intake including vegetable and milk production could be important factor for improving learning motivation in students.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Studying Stimulus‐Induced Changes in RNA Dynamics by Mutational Mapping through TimeLapse‐seq
Authors: Kiefer Lea , Schofield Jeremy A , Simon Matthew D
Published: June 2021

The eukaryotic transcriptome is dynamically regulated through RNA transcription and decay allowing cells to respond to environmental stimuli in a rapid and specific manner. The bromodomain inhibitor JQ1, has been extensively studied in the context of cancer cells, as it blocks the activation of specific sets of oncogenes perturbing RNA population dynamics.1 Recently, JQ1 has additionally been found to repress pathogenic response genes in models of heart failure, however the mechanism of this action remains unclear.2 JQ1 is known to block the BET bromodomain reader proteins involved in chromatin remodeling, affecting the transcription of the target genes. We, therefore, aim to study the effect of JQ1 in heart failure on the transcriptional level. While RNA‐sequencing (RNA‐seq) provides a snapshot of the cellular transcriptome, temporal information is lost as rapid changes in the RNA population remain undetected among the excess of preexisting RNA. To gain insight into the role of JQ1‐induced pathways in cardiomyocytes, we are studying changes in RNA dynamics using TimeLapse‐seq3, recently developed nucleotide recoding chemistry that utilizes metabolic labeling in an enrichment‐free RNA‐seq approach. Through recoding of the hydrogen bonding pattern of the metabolic label, TimeLapse‐seq identifies new transcripts through apparent mutations in the RNA‐seq reads. We will describe progress using TimeLapse‐seq as a strategy to study differently regulated transcripts in disease and to deduce the mechanism of JQ1 in heart failure.

Support or Funding Information

This work was supported by the NIH NIGMS T32GM007223 (J.A.S. and E.E.D); NSF Graduate Research Fellowship (E.E.D); NIH New Innovator Award DP2 HD083992‐01 (M.D.S.), and a Searle scholarship (M.D.S.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ventricular activation times in rainbow trout and common carp
Authors: Vaikshnoraite Marina Alvirasovna , Azarov Jan
Published: June 2021

The specific objective of the present study was to find out the presence or absence of a transmural gradient of ventricular activation times in fish having pyramidal and sac‐like hearts, which in turn have relatively pronounced and negligible amount of the compact type of myocardium, respectively.

Experiments were done in rainbow trout (Onchorhynchus mykiss, n=6; body weight 320–510 g) and common carp (Cyprinus carpio, n=6; body weight 900–1300 g) presenting fish species with the pyramidal (pronounced compact layer) and sac‐like (negligible compact layer) heart types. The investigation was carried out in accordance with the Guide for the Care and Use of Laboratory Animals, 8th Edition published by the National Academies Press (US) 2011. Body surface ECG was recorded in the longitudinal body axis lead. Ventricular unipolar electrograms were recorded from the ventricular walls as described earlier (Vaykshnorayte et al., 2011). In each ventricular lead, activation time were determined as instants of dV/dt minimum during QRS complex (Coronel et al., 2006). Data are expressed as medians and interquartile intervals. Statistical analysis was performed with SPSS package (IBM SPSS Statistics 23, SPSS, Inc., Chicago, Illinois, USA).

Common carps demonstrated significantly longer QRS complex in comparison with rainbow trout (79 (IQR 72–85); 56 (IQR 48–61), P<0.05), whereas a difference in RR interval was statistically insignificant (1892 (IQR 1609–2074); 1470 (IQR 970–2175), P>0.05). The gross activation sequence in both species was similar and demonstrated a direction from atrioventricular junction to apex and anterior base (apicobasal activation gradient, P <0.005). The transmural activation pattern was also similar in the studied animals with the activation spread direction from endocardium to epicardium in all parts of the myocardium (transmural activation gradient, P <0.005). In this way, It implies that a transmural activation pattern do not have differences in hearts with different spongy/compact percentage of the ventricular wall in these fish species.

Support or Funding Information

This work was supported by the Ural Branch of the Russian Academy of Sciences (AAAA‐A18‐ 118012290365‐2) and the Program for Fundamental Research of RAS (AAAA‐A17‐ 117012310154‐6).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Precision Medicine in Psychiatry by Predicting Lithium Treatment Response using Gene Expression Biomarkers and Machine Learning
Authors: Eugene Andy Roger , Eugene Beata , Masiak Jolanta
Published: June 2021

Background

We tested the hypothesis that transcriptional‐level genes signatures are differentially expressed between male and female bipolar patients, prior to lithium treatment, in a patient cohort who were later were clinically classified as lithium treatment responders using the primary outcome measure of the Clinical Global Impression Scale for Bipolar Disorder‐Severity (CGI‐BP‐S).

Methods

Gene expression study data was obtained from the Lithium Treatment‐Moderate dose Use Study data accessed from the National Center for Biotechnology Information's Gene Expression Omnibus via accession number GSE4548. Differential gene expression analysis using the CGI‐BP‐S responder status and Gender served as control variables for analysis using the Linear Models for Microarray and RNA‐Seq (limma) package and the Random Forests machine learning algorithm in R for Statistical Computing programming language software.

Results

In pre‐treatment lithium responders, the following genes were found having a greater than 0.5 fold‐change and differentially expressed indicating a male bias: RBPMS2, SIDT2, CDH23, LILRA5, and KIR2DS5 while the female‐biased genes were identified as: HLA‐H, RPS23, FHL3, RPL10A, NBPF14, PSTPIP2, FAM117B, CHST7, and ABRACL.

Conclusions

Applying machine learning methods to gene expression data, we developed a pre‐treatment gender‐specific gene‐expression‐based predictive model selective for lithium treatment responders according to the Clinical Global Impression Scale for Bipolar Disorder‐Severity with an ROC AUC of 0.92 for men and an ROC AUC of 1 for women.

Support or Funding Information

None.
Low‐Flow‐Mediated Constriction is Reduced in Healthy Postmenopausal Women
Authors: Kuczmarski Andrew , Sebzda Kelly , Wenner Megan
Published: June 2021

Vascular function declines in women after menopause, and has been characterized by a reduction in brachial artery flow‐mediated dilation (FMD). Low‐flow‐mediated constriction (L‐FMC) is an assessment of resting arterial tone that serves as a complementary measure of vascular function. Although L‐FMC has been shown to be reduced in certain diseased populations such as coronary artery disease, to our knowledge it is unknown if L‐FMC is altered with age/menopause. Accordingly, the purpose of this study was to test the hypothesis that postmenopausal women (PMW) have reduced L‐FMC values compared to young women (YW). We assessed vascular function in 16 PMW (59±1 years, 24±1 kg/m2) and 19 YW (21±1 years, 23±1 kg/m2). Longitudinal images of the brachial artery were acquired using ultrasound (GE Logic P5). A blood pressure cuff was placed just distal to the olecranon process and inflated to 200mmHg for five minutes. Images were recorded continuously during baseline (1‐minute), cuff occlusion (5‐ minutes), and cuff release (2‐minutes). L‐FMC was calculated as a percent change from baseline diameter to the average diameter during the last 30 seconds of cuff occlusion. FMD was calculated as a percent change from baseline diameter to peak diameter following cuff release. Total vessel reactivity was calculated as the sum of the absolute value of L‐FMC and FMD. All women were normotensive, although PMW had higher resting mean arterial pressure (PMW: 90±2 mmHg, YW: 82±2 mmHg; P<0.01). L‐FMC was reduced in PMW compared to YW (PMW: −1.67±0.36% vs YW: −2.84±0.35%; P=0.03). FMD was lower in PMW compared to YW (PMW: 5.47±0.54% vs YW: 7.33±0.53%; P=0.02). Total vessel reactivity was also lower in PMW (PMW: 7.32±0.67% vs YW: 10.17±0.57%; P<0.01). Consistent with previous findings, PMW have lower FMD values compared to YW. In addition, PMW also have reduced L‐FMC compared to YW. These findings suggest that L‐FMC may be a useful measure of vascular function with healthy aging.

Support or Funding Information

Supported by: NIH Grant P20 GM 103446, U54 GM 104941, P20 GM113125, and University of Delaware 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.