bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolic Markers of Hyperhomocysteinemia in Healthy Adults Population
Authors: Waly Mostafa I. , Ali Amanat , Padmanabhan Smitha , Al‐Farsi Yahya 
Published: June 2021

BackgroundHyperhomocysteinemia is involved in the pathogenesis of oxidative stressmediated human chronic diseases including; cancer, cardiovascular diseases, hypertension, and type 2 diabetes. B vitamins (folate, vitamin B6, and vitamin B12) deficiencies induce hyperhomocysteinemia, and thereby contributes to the pathology of human chronic diseases.ObjectiveThis study aimed to assess the biochemical and nutritional status of B vitamins in relation to hyperhomocysteinemia among healthy adults population.MethodsThe study participants (n=400; 190 males and 210 females) were healthy adults subjects and free of any chronic diseases or endocrine disorders. They were supplied with B vitamins for 6 months and the homocysteine and B vitamins levels were measured twice in the sera of the study subjects (before starting the trial and at the end of the trial). The nutritional status was assessed using prospective semi‐quantitative food frequency questionnaire.ResultsLow B vitamins levels were observed in the sera of study participants with hyperhomocysteinemia (fasting serum homocysteine >15 μmol/l). Meanwhile, B vitamins supplementation was effective in combating hyperhomocysteinemia and its associated oxidative stress indices as evidenced by enzymatic and non‐enzymatic antioxidants markers.ConclusionTaking adults population with hyperhomocysteinemia as a high risk group; the results of this study indicate that B vitamins supplementation modulates hyperhomocysteinemia‐mediated oxidative stress. Serum B vitamins levels might be used as a novel biomarker for early diagnosis and screening of hyperhomocysteinemia,Support or Funding InformationThis research was supported by Internal Grant Funding (IG/AGR/FOOD/17/1), College of Agricultural and Marine Sciences, Sultan Qaboos UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigating the Role of FLO1 in the Lack of Flocculation of Yeast Brewing Strain Thirsty Pagan Local
Authors: Rieffer Amanda Elizabeth , Via Zachary 
Published: June 2021

Brewer's yeast use a fermentation pathway to convert sugars into ethanol in the beer making process. An interesting characteristic most yeast have is the ability to clump together, known as flocculation. Flocculation is useful for brewing companies because it makes it much simpler to separate and reuse it for later brews. Our research is focused on a locally found strain of Saccharomyces cerevisiae, Thirsty Pagan Local (TPL), that does not flocculate, and we want to know why. There are a series of genes associated with flocculation, and FLO1 codes for a protein on the surface of the cell that aids in flocculation. An ethanol sensitivity assay was conducted, and based on this know that FLO1 for TPL is not a null mutant, so it's possible that this gene is simply under expressed. When comparing TPL FLO1 with wild type DL1 FLO1 (a known nonflocculator), we noticed that our FLO1 is roughly 1,000 base pairs shorter than wild type. We are in the process of sequencing TPL FLO1 to find areas where there is a detrimental amino acid change that could give us a clue as to why our strain doesn't flocculate. We have successfully inserted FLO1 into an overexpression plasmid, and plan on conducting a flocculation assay with this plasmid along with FLO1 from a known flocculator, 1195. If the overexpression of TPL FLO1 results in flocculation, our next plan is to establish the overexpression vector permanently into the yeast genome.Support or Funding InformationThe College of St. ScholasticaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Impact of the Physical Micro‐Environment on Notch Signaling
Authors: Detweiler Michael Allen 
Published: June 2021

Notch is a transmembrane ligand receptor that is involved in cell‐cell communication via canonical binding to a mounted ligand on an adjacent cell. Upon activation of ligand binding, Notch's intracellular portion is cleaved and translocates to the nucleus to function as a transcriptional regulator of many downstream genes. Extracellular micro‐environmental cues including shear stress, ECM stiffness, pH, etc. have been shown to impact Notch activation. Integrins are dimerizing proteins that aid in attachment of the cell to the extracellular matrix, which can have impacts on cell physiology from the micro‐environment. Recently published data has also revealed that integrins have an effect on Notch activation, but the mechanism(s) inducing this response have yet to be discovered. Because the extracellular micro‐environments can impact cell physiology through integrin functions, we hypothesize that these physical micro‐environmental conditions may also impact Notch signaling. To investigate this hypothesis, we monitored Notch activation during shear stress or increasing extracellular matrix stiffness, two conditions shown to influence integrin downstream signaling. Our preliminary results show that applying shear stress on endothelial cells provides an increase in downstream Notch signaling activity, which is effected by inhibiting integrin‐linked molecules. Our preliminary results also suggest that Notch activity is controlled by matrix stiffness and that inhibiting Notch activation on stiff extracellular matrix impacts the production of smooth muscle actin (indicator of fibrosis), suggesting a pathway to fibrosis through an integrin‐Notch connection. Together, these experiments support our hypothesis of the impact of the physical microenvironment on Notch signaling through integrin activation. By coming to these conclusions, we hope to gain a better understanding of the impacts integrins and the microenvironment have on Notch signaling and to identify the mechanisms involved.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison Between the Structure‐Function Relationships in Oncogenic and Wild‐Type Gα Subunits
Authors: Goossens Jesse , Leverson Brian , Freitas Duarte 
Published: June 2021

Guanine nucleotide binding proteins (G‐proteins), in conjunction with their receptors (GPCR), are one of the most prevalent signaling systems and are involved in the regulation of an array of cellular processes. Upon binding of an extracellular stimulus to the GPCR, the activated α‐subunit (Gα) exchanges GDP for GTP, releases both the βγ‐complex and the GPCR, and associates with an effector that further relays the signal. The signal terminates with the hydrolysis of GTP to GDP, causing the α‐subunit to reunite with the βγ‐subunits and the GPCR. G‐protein signaling must be tightly regulated to ensure the appropriate responses to extracellular stimuli. Any misregulation caused by mutations can be detrimental. Gsα and Giα1 are G‐protein subunits that, respectively, stimulate or inhibit adenylyl cyclase and regulate the production of the secondary messenger, cAMP. We studied two mutations observed in cancers of the intestine (R231H mutation in Gsα) and the colon (R208Q mutation in Giα1). Spectroscopic and crystallographic approaches were used to further understand these disease states.Fluorescence spectroscopy, circular dichroism, and UV/Vis spectroscopy showed lower melting temperature in the active conformation of the oncogenic mutants, indicating disruptions in the noncovalent interactions. Using malachite green and fluorescence GTP assays, we found that the kcat values for oncogenic Giα1 were lower than for the WT protein; the trend was the opposite for the WT and oncogenic Gsα proteins. When comparing the x‐ray structure of the R208Q mutant with that of the WT Giα1, the oncogenic point mutations do not show significant differences in the structure of the protein as a whole. However, molecular dynamics simulations modeling the interactions of T181 in Giα1 and T204 in Gsα (key residues in the hydrolysis of GTP) show drastically different interaction energies between mutants and WT proteins. Taken together, these findings suggest that the altered rates of GTP hydrolysis lead to an imbalance of cAMP in tumor cells.Support or Funding InformationNIH R15GM112025This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elucidating structure‐function relationships of the human protein kinase MEK1
Authors: Lee Rebecca T. , Ravatt Leandre M. , Runco Caroline E. , Oza Javin P. 
Published: June 2021

MEK1 is a dual specificity protein kinase in the MAPK phosphorylation cascade. As one of the prevalent signal transduction pathways, the MAPK pathway regulates a diverse variety of cellular processes including proliferation, differentiation, and apoptosis. The phosphorylation event between MEK1 and its only known substrate ERK2 acts as regulatory bottleneck for signal transduction, and maintains the fidelity of signals coming in and going out. As such, MEK1 remains a target for therapeutic intervention of a variety of cancers. While much is known about MEK1's role in cell signaling, enzymatic characterization of MEK1 has been limited due to difficulty in obtaining the active, doubly‐phosphorylated MEK1 in a homogenous population. Our study overcomes this barrier by utilizing an expanded genetic code for the co‐translational incorporation of phosphoserine into MEK1. Specifically, we are leveraging this capacity for on‐demand production of site‐specifically phosphorylated MEK1 to characterize the human MEK1 and elucidate the structure‐function relationships that governs MEK1 activity. Using a hypothesis‐driven approach based on conserved kinase motifs, we selected residues within the activation segment and catalytic loop for site‐specific mutagenesis. Here we report our efforts to express and characterize the phosphorylated MEK1 and its mutants through immunoassays for a comparative analysis of kinase activity. Our work aims to establish kinetic parameters for the human MEK1 for the first time and provide insights into mechanisms of substrate binding and catalysis.Support or Funding InformationOur research is funded by the Center for Applications in Biotechnology @ Cal Poly and the Bill and Linda Frost Fund.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein Kinase Cα (PKCα) Gain‐of‐Function Variant in Alzheimer's Disease Displays Enhanced Catalysis by a Mechanism that Evades Down‐Regulation
Authors: Callender Julia , Yang Yimin , Stephenson Natalie , Jones Alexander , Brognard John , Newton Alexandra 
Published: June 2021

The signaling output of protein kinase C (PKC) is precisely controlled through multiple mechanisms that balance its activity in order to maintain cellular homeostasis, with disruption of this balance resulting in pathophysiologies. In this vein, the cell has evolved mechanisms to maintain this balance by degrading constitutively active PKCα. We have previously shown that loss‐of‐function mutations in multiple PKC isozymes contribute to cancer, a disease of aberrant proliferation, and gain‐of‐function mutations in the conventional isozyme PKCα contribute to Alzheimer's Disease (AD), a disease of aberrant cell death. Here, we biochemically characterize one of the AD‐associated variants, PKCα‐M489V, in order to investigate the mechanism through which it confers enhanced signaling output without causing PKCα degradation. We use a combination of in vitro and in silico methods to demonstrate that this mutation effectively increases the specific activity of purified PKCα under activating conditions by increasing the molecular dynamics of the catalytic domain. Importantly, it does so without affecting the normal autoinhibition of basal signaling, thereby allowing for an enhanced signaling output that simultaneously evades the cell's homeostatic degradation of PKCα protein levels. Taken together, our results provide an in‐depth characterization of a disease‐associated gain‐of‐function PKC mutation, and for the first time report a mutation that enhances signaling output of PKCα while maintaining its stability in the cell.Support or Funding InformationThis work was supported by NIH R35 GM122523 and NIH GM43154 (ACN), the Cure Alzheimer's Fund (ACN), and Cancer Research UK (NS, JB). JC was supported by the UCSD Graduate Training Program in Cellular and Molecular Pharmacology (T32 GM007752).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enhancing Scientific Communication About and Ethical Reasoning Surrounding Gene Modifications Through a Collaborative Learning Experience
Authors: Danowitz Amy M. , Ehret Verna M. 
Published: June 2021

Scientific communication and the ability to think about the ethical impact if one's work are necessary skills for all scientists to obtain. These are especially important skills for scientists working in cutting edge fields that have the potential for major societal implications, including scientists working with genetic modifications of humans and animals. Unfortunately, the traditional undergraduate curriculum generally does not provide many opportunities for students to practice communicating their science to the general public, nor does it offer much ethical training beyond standard responsible conduct of research requirements. Herein, we describe a novel collaborative learning experience that allows students who are studying modern genetic modification techniques (including traditional methods of gene modification, as well as newer advances such as RNAi and CRISPR/cas9) to work with students who are studying ethical reasoning. As part of this collaboration, the science students are required to hone their understanding of a technique used in genetic modification and to explain this technique to a largely non‐scientific audience (the ethics students). The ethics students, in turn, teach the science students about some of the ethical concerns that can arise around these techniques. By participating in this experience, students gain a better understanding of how to best communicate complex, technical information with non‐scientists and a fuller idea of how ethical decision making can impact their lives as laboratory scientists and as citizens of the world. Both the faculty and the students involved in this collaboration found this to be a worthwhile and enjoyable experience. The faculty were particularly impressed by the high levels of student work and thought put into this project. This poster will address the assignments that were given, the grading standards for these assignments, general student opinions on this project, and ideas for where and when to fit such collaborative projects into the undergraduate curriculum.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 Toxic Collaboration: Using SPIONS and Shrimp to Bridge Courses in Chemistry and Biology for Undergraduate Engineering Students.
Authors: Rogers Ryan Patricia , Grove Laurie Eileen 
Published: June 2021

Chemistry and biology are naturally symbiotic disciplines, which provide foundation for other related areas such as engineering. At Wentworth Institute of Technology, the majority of students enroll in our Engineering Chemistry course and many others will also take Cellular and Molecular Biology, either to fulfill a major requirement or science elective. It is between these two courses that we have designed a collaborative project investigating the synthesis and biological relevance of superparamagnetic iron oxide nanoparticles (SPIONs). SPIONs are at the forefront of investigation across several different fields, which makes them an exceptional interdisciplinary educational tool. This project specifically focuses on the role of SPIONs as contrast agents for MRI, which directly relates to future coursework for our biomedical and biological engineering majors; however, the construction of the NMR and MRI instruments, including the superconducting coil, provides interest for our electrical and mechanical engineering students. Students in chemistry are paired with cohorts of biology students and directed to compile their experimental data to generate a complete narrative related to the synthesis and uptake of SPIONs by brine shrimp. Numerous synthetic variations of SPIONs are available, thus allowing students to develop and test a hypothesis relating to either the magnetic properties or toxicity of the nanoparticles, which can only be explained and supported at the conclusion of both experiments.The collaboration culminates with a video lab report, developed and presented by both cohorts of students. Scores are generated based on presentation style and creativity, scientific accuracy, ability to develop and explain their hypothesis, data analysis and interpretation of findings on a larger scale. The collaboration takes place over the course of 3 2‐hour lab periods, with additional check points during the biology component. Two of these periods are designated for synthesis and characterization of SPIONs by the chemistry students and one lab period is designated for induction of toxicity assays and cytotoxicity analysis by the biology students.Support or Funding InformationThis work was supported by the Department of Sciences at Wentworth Institute of Technology.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Paclitaxel on Glucocorticoid‐induced apoptosis of Human Leukemic CEM Cells
Authors: Aranas Ivymarie , Medh Rheem 
Published: June 2021

Glucocorticoids (GCs) are steroid hormones widely used in anti‐leukemia therapy as they can induce apoptosis through certain gene‐regulatory pathways that affect pro‐ and anti‐apoptotic genes. In cell culture models of human acute lymphoblastic leukemia (ALL), the synthetic GC, Dexamethasone (Dex), has been shown to induce expression of pro‐apoptotic genes including E4BP4 (a transcription factor that upregulates pro‐apoptotic BIM), and regulators of apoptosis such as FOXO3 and BIRC3, in correlation with apoptosis. Paclitaxel (Ptx), a taxol derived anti‐proliferative alkaloid, induces apoptosis via a distinct but overlapping transcriptional response, and is often used in combination with Dex as an anti‐leukemic agent. In this study we are investigating the effect of Ptx alone, and in combination with Dex, on apoptosis and regulation of key pro‐ and anti‐apoptotic genes, in an effort to understand the molecular implications of combined treatment with Dex and Ptx. The CCRF‐CEM cell lines CEM‐C7‐14 and CEM‐C1‐15 were previously found to be sensitive and resistant, respectively, to Dex. Sensitivity to Dex was restored in CEM‐C1‐15 cells upon ectopic expression of mouse E4BP4 (line CEM‐C1‐15mE#3). We are testing the hypothesis that E4BP4 and BIM play a central role in regulating apoptosis triggered by Ptx as well, and that combined treatment synergistically triggers apoptosis via a greater magnitude of upregulation, in addition to activating a broader spectrum of pro‐apoptotic pathways. All three CEM cell lines were treated with 1 mM Dex and 1mM Ptx either alone or in combination for 24–96h. All three cell lines were sensitive to Ptx, but CEM‐C1‐15mE#3 was least susceptible. Apoptosis was confirmed by staining cells with Annexin V‐FITC followed by epifluorescence microscopy. Reverse transcription and end‐point PCR followed by ImageJ analysis was employed to evaluate relative gene expression in response to each treatment. Our data suggest that Ptx upregulated FOXO3, BIRC3, and BIM, but down‐regulated E4BP4 in the CEM‐C7‐14 cells, suggesting that Dex and Ptx have distinct effects on E4BP4 regulation but may share common downstream pathways for apoptosis via BIM upregulation.Support or Funding InformationNIH SC3 grant, #GM081099, and a Building Infrastructure Leading to Diversity (BUILD) grant from the NIH, #5RL5MD009603This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Primary Acute Lymphoblastic Leukemia Cells Strictly Require Interphase Microtubules for Survival during G1 Phase Advance
Authors: Delgado Magdalena , Chambers Timothy C. 
Published: June 2021

Microtubule targeting agents (MTAs), such as vincristine (VCR), are important anticancer agents and are thought to act primarily in mitosis. However, there is increasing evidence that MTAs can also cause death by interfering with interphase microtubules. We have recently discovered that when primary acute lymphoblastic leukemia (ALL) cells are treated with vincristine, cells in G1 phase die directly without advancing to other phases of the cell cycle. This system represents a powerful model for understanding interphase mechanisms of MTA action, and provides an opportunity to test whether MTAs can be combined with agents that target interphase processes in order to create novel treatments for ALL. One such candidate is palbociclib (PCB), a highly selective inhibitor of cyclin dependent kinases 4/6 (CDK4/6), whose activity is necessary for the transition from G1 to S phase, providing a rationale for the combination of palbociclib and vincristine to target ALL cells in G1 phase. In this study, we tested the effect of PCB alone or in combination with vincristine in primary ALL cells. PCB caused arrest of primary ALL cells in G1 phase, with 97–99% of cells exhibiting 2N DNA content, while having no significant effect on cell viability. In contrast, HeLa cells were unaffected by PCB, consistent with their lack of dependence on the CDK4/6‐retinoblastoma pathway. When ALL cells were pretreated with PCB, they became refractory to VCR treatment. Examination of the microtubule network by immunofluorescence microscopy in ALL cells treated with PCB alone or PCB followed by VCR treatment showed that PCB did not disrupt the microtubule network, nor prevented VCR from doing so. Furthermore, ALL cells treated with ABT‐263, a Bcl‐2/Bcl‐xl inhibitor, after PCB pretreatment still underwent cell death, indicating that PCB did not block downstream apoptotic pathways. Experiments assessing entry and passage through S phase using 5‐ethynyl‐2′‐deoxyuridine incorporation showed that G1 arrest in primary ALL cells by PCB was reversible, and that cells released from arrest regained susceptibility to VCR. Taken together, these results indicate that ALL cells must be cycling in order for interphase death induced by vincristine to occur, and in turn indicate that interphase microtubules play a fundamental role in survival of actively cycling ALL cells. In addition, these findings strongly caution against combining vincristine and CDK4/6 inhibition as a potential therapy for ALL.Support or Funding InformationChancellor's Circle AwardThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.