bluuebunny/crossref_metadata_2025_split huggingface dataset:
ATP as an Allosteric Modulator and Chelator of Fe3+ from Fe3+‐transferrin. Is ATP a Major Fe3+ Carrier
Authors: Cowart Richard E. , Shaffer Tyler B. , O'Hara Ryan D. , Campbell Jacquelyn A. , Schneider‐Tugan Lori L. , Binz Sara K. , Pope Mitchell A. , Gregory Robert B. 
Published: June 2021

The objectives of this study were to examine two chelators in the removal of Fe3+ from Fe3+‐transferrin. One was pyrophosphate (PPi) as it is a reasonably effective chelator in this reaction, and the other was ATP in that it consists of PPi attached to adenosine monophosphate. We reasoned that ATP might bind to a nucleotide binding site if present and in close proximity to the metal center, to enhance Fe3+ removal and give insight into the mechanism of Fe3+ release. In this study we used uv/vis spectroscopy to examine the removal of Fe3+ from Fe3+‐transferrin by time‐course and Vi reactions. At acidic pH we determined association constants of Fe3+‐transferrin alone, and Fe3+‐transferrin in the presence of ATP. We also employed molecular modeling to identify and characterize a putative ATP binding site. Our results show that sigmoidal kinetics were obtained as a function of pH when ATP was the chelator. At pH 7.5 the kinetic data showed there was no removal of Fe3+ by ATP. ATP was found to exert positive heterotropic effects at acidic pH and resulted in the increase in the association constant of Fe3+‐transferrin. These effects were not seen with PPi. We concluded that there were two destabilizations of Fe3+‐transferrin necessary for Fe3+ release; the first due to a low pH induced conformational change in the protein and the second due to ATP, also at low pH, where ATP served as an allosteric modulator. Essentially 100% of Fe3+ was released from both the N‐ and C‐lobes at pH 5.0 in the presence of ATP in ~ 3 minutes as determined by time‐course reaction. Molecular modeling studies identified a putative 25 amino acid ATP binding channel in the N‐lobe of human serum transferrin. These conserved amino acids were identified on each open face of the NI/NII subdomains of apo‐transferrin with the channel forming upon NI/NII subdomain closure as Fe3+ is acquired. At neutral pH ATP bound approximately 9 Å from Fe3+ in the metal center. At acidic pH the protein conformational change positioned ATP immediately adjacent to Fe3+ for rapid chelation. In that ATP is an extracellular, allosteric signaling molecule a mechanism of Fe3+ release from Fe3+‐transferrin is proposed. Here, extracellular ATP is acquired by transferrin in the serum at neutral pH where chelation does not occur. Upon binding to the transferrin receptor and internalization into an acidic endosome, a pH induced conformational change along with positive heterotropic effects occur to allow ATP to efficiently chelate Fe3+. We conclude that transferrin is an allosteric protein, ATP is an allosteric modulator and effective chelator, and that ATP is likely a major Fe3+ carrier in higher organisms.Support or Funding InformationState of Iowa grant GIVFThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identifying Substandard Medications in Developing Nations
Authors: Omanovich Esad , Donels Jordan , Trier Justin , Thomsen Alli , Dollen Julia , Miller Austin , Memic Nejla , Zea Corbin 
Published: June 2021

Access to adequate and quality health care is a worldwide concern. Without proper access to medicine individuals will often suffer diseases that could be treated or prevented with the proper medicine. Proper dosage is also necessary so that patients can recover from the disease as well as stop the production of drug resistant bacteria and parasites. In collaboration with Norte Dame University, a protocol, using High Performance Liquid Chromatography (HPLC), was developed to quantify the amount of Albendazole, a deworming medication, in tablets from locations in Africa to ensure proper dosage. A Spike and Recovery, a secondary verification method, was developed to verify the efficiency of the column and protocol used. Optimization of the Spike and Recovery method was completed and showed that the efficiency of the column was 97 ± 1 %. Using this protocol, multiple samples have been quantified with one sample falling below the advertised dosage.Support or Funding InformationGrow Iowa Values FundThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Light Controlled Intracellular Protein Release: Tracking Ras Interactions With Superresolution Fluorescence Microscopy
Authors: Yun Jason , Phelps Carey , Morales Demosthenes , Nan Xiaolin , Reich Norbert 
Published: June 2021

Although fusion proteins have been extremely helpful in the study of intracellular protein dynamics, this approach has significant drawbacks. We circumvent such problems by delivering dye‐conjugated proteins using our Nano‐optogenetic platform, which relies on hollow gold nanoshells to release proteins with NIR laser exposure. Herein, we report the delivery of a NS1 monobody, known to prevent dimerization of RAS GTPases that inhibits downstream signal transduction for cell division. Our modular nanoplatform is appended with thiol linked scaffolds containing a streptavidin‐biotin cell‐penetrating peptide for cell internalization and a nitriloacetic acid moiety to bind the polyhistidine dye‐labeled NS1 in the presence of copper. Near‐infrared laser activation allows for controlled spatio‐temporal endosomal release of the dye‐labeled NS1 to colocalize with RAS GTPase. Our approach can deliver any dye‐label His‐tagged protein of interest with controlled spatio‐temporal release inside cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Creation and Characterization of Gold‐Nanoparticle Containing Conductive Scaffolds for Culturing Cardiomyocytes
Authors: Maldonado Marcos , Castellanos Brisa Pena , Bonham Andrew J. , Park Dae Won 
Published: June 2021

Cardiomyopathies, diseases of the heart, are one of the major causes of death in the United States, and thus there is great interest in preventing and treating these complications. However, due to the nature of limited availability of donors, many techniques and solutions are inadequate to meet the needs of the field. In particular, heart tissue transplantation, culturing human tissues/tissue‐derived cells and tissue engineering, and finding a suitable extracellular environment that closely resembles the natural environment of cardiomyocytes in vivo are difficult to attain. As such, a great deal of work has gone into efforts to produce polymers which mimic the natural cell environment in properties such as binding sites, stiffness, reactivity, and hydration. In our research, we are investigating the development and characterization of conductive polymer scaffolds providing cardiac tissue support, which will ideally aid in culturing cardiomyocytes for academic, research, and medical use. The core of our model is the incorporation of conductive gold nanorods into reverse thermal gel polymers. Synthesized gold nanorods utilized in this procedure are made with a high aspect ratio, at high purity and with defined surface functionalization. These conducting scaffolds should properly accommodate cardiac cells in building functional cardiac tissue constructs and further improving cell retention, spreading, cardiomyocytes long term survival and maturation, cell‐cell coupling and synchronized beating behavior at the tissue level. The nanoparticles have been synthesized, then covalently coupled into these scaffolds, resulting in order‐of‐magnitude increases in conductivity. The goal of this work aims to improve cardiac tissue engineering, so that it can be directed to ultimately repairing damaged heart muscle and improve overall cardiac function in cardiovascular diseases whether they have been acquired from past medical complications or developed through hereditary traits.Support or Funding InformationThis work is supported by internal grant funding from the Metropolitan State University of Denver.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
iTraq‐based Proteomics analysis of colon mucosal proteins in a dextran sulfate sodium (DSS)‐induced colitis mouse model and the effects of dietary treatments with edible mushroom Pleurotus eryngii
Authors: YUAN BIAO , XU XIAO , HAN YANHUI , CAO XIAOQIONG , HU QIUHUI , HANG XIAO 
Published: June 2021

Our previous studies have demonstrated the protective effects of an edible mushroom Pleurotus eryngii against the colitis in mice. Herein, an itraq‐based proteomics analysis has been utilized to establish the potential mechanism of actions of the mushroom. Three groups of mice (10 mice per group) were treated with DSS in drinking water (the colitic positive control group), DSS in drinking water plus Pleurotus eryngii (1.5% in diet, the treatment group), or regular drinking water (the negative control group), respectively, for 40 days as we described previously. The colon mucosa of mice were collected and subjected to an itraq‐based proteomic analysis. In total, 5794 proteins were identified by LC‐ESI‐MS/MS analysis in the colon mucosal of the mice. Among them, the expression levels of 626 proteins were different (392 up‐regulated proteins and 234 down‐regulated proteins) in the colitic positive control mice in comparison with the noncolitic negative control mice. There were 303 differentially expressed proteins (107 up‐regulated and 196 down‐regulated) in the treatment group compared to that of the positive control t group. The KEGG pathways that these differentially expressed proteins involved in were mainly: Steroid hormone, Arachidonic acid, Basal cell carcinoma, Linoleic acid metabolism, Phospholipase D signaling, viral myocarditis, chemical carcinogenesis, natural killer cell mediated, thyroid hormone signaling pathways. Twenty‐five proteins that were highly upregulated in the colitic positive control group (vs. the negative control group) were found significantly down‐regulated in the treatment group (vs. the colitic positive control group), whereas, twenty‐one proteins that were highly down‐regulated in the colitic positive control group (vs. the negative control group) were found significantly up‐regulated in the treatment group (vs. the colitic positive control group). Importantly, western blot analysis validated the changes in the protein expression described above. These results demonstrated that dietary mushroom treatment reversed, at least partially, the abnormal expression of proteins in the colon mucosa of colitic mice, which may be accounted for the protective effects of Pleurotus eryngii mushroom against pathogenesis of colitis in mice. Overall, our results provided preliminary but critical information on the potential mechanism of Pleurotus eryngii mushroom in inhibiting colitis.Support or Funding InformationThis research work was funded by the National Natural Science Foundation of China under Grant No. 31471927.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Integrated Therapeutic Dynamics by Applications of Multi‐Mathematical Functions in Multiple Degenerative Chronic Physiological Processes. A Case study
Authors: Einstein George P. , Tulp Orien L. , Konyk Carla M. , Kealoha Nicholas P. 
Published: June 2021

BackgroundMost chronic diseases and conditions affect one or more organs. The degeneration process overlap in chronic diseases and is very difficult to treat. Nonetheless, 7 of every 10 Americans who die each year—more than 1.7 million people—succumb to a chronic disease. Health‐damaging behaviors, such as tobacco use, lack of physical activity, poor eating habits, and excessive alcohol use contribute to many chronic diseases, whereas others may represent the long‐term effect of early exposure to toxins and/or other environmental factors, especially in individuals with a higher genetic risk of disease. A shared aspect of many chronic diseases is chronic pain and other disease‐associated disability that interferes with quality of life. Both psychological factors and a general physiological medical condition are judged to have important roles in the onset, severity, exacerbation, or maintenance of the pain.Materials and MethodsPatient suffering forty‐three years in chronic pain and countless medical diagnosis including COPD, Diabetes, MRSA, Irritable Bowel Syndrome, Fibromyalgia, Cronic Renal Failure, Ulcerative Cortices and Major Depressive Disorder. Patient was barely able to walk, with Multiple physio‐pathological processes of chronic diseases, and required a complex design to bring about a reversal therapeutic process. The Multi‐Mathematical Functions in Multiple Degenerative Chronic Physiological Processes (MMFPP) are represented by FN Specific Coherencies Therapeutic Functions and Specific Therapeutic (M} MatricesResultsBrain activity was essential for Diagnostic Multilevel Physio Pathological processes. Integrative therapy by Specific Dynamic Matrixes of Sigma Quanta Energy by Biophysical‐Biochemical and Psycho‐neural Stimulation was effective in amelioration of symptoms.Support or Funding InformationSupported by Institutional Resources of USAT Montserrat and the Einstein Medical Institute, NPB, FLThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Low pH Treatment on Cell Cycle and Cell Growth
Authors: Hu Yuli , Li Yang 
Published: June 2021

Cancer growth affects the pH of microenvironment that becomes acidified and supports local invasion. Studies have reported that pH is a factor in cancer growth, division, and spread. Although cancer cells produce low extracellular pH (6.5–6.9), they are able to maintain their intracellular pH at favorable ranges (7.2–7.4). Our preliminary results showed that growth of cancer cells changes after low pH treatment. The objective of our research is to study how low pH affects cell morphology and behavior. In my preliminary experiments, HeLa cells were incubated with low pH solution for 10 min and harvested using trypsin into medium with normal pH. After 24 hours in culture, cell number was assessed using Trypan Blue Exclusion Assay. As a result, low pH treated group doubled 1.4 times; whereas control group doubled 2.3 times, suggesting a division latency in low pH treated group on Day 1. Cell morphology on Day 1 was recorded as images and was categorized into round, spindle, triangle, and irregular groups, among which spindle cells were likely to be at G1 phase. Percentage of the spindle cells gradually increased during 16–24 hours in experimental group; however, in control group, percentage of spindle cells decreased, suggesting that low pH had induced cell cycle arrest at G1 phase. In conclusion, low pH treatment affects cell growth potentially through G1 cell cycle arrest. Future work is to confirm cell cycle arrest at G1 phase after low pH treatment and to study the mechanism of low pH effect on cancer cell growth.Support or Funding InformationFunding is provided partially by NIH (#NS081629) to YVL. Thanks to the Osteopathic Heritage Foundations, Graduate Assistantship Program at Ohio University Heritage College of Osteopathic Medicine for supporting graduate student YH.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
DMD‐10 and its Potential Effect on Regulation of the Glutamate Receptor GLR‐1 in C. elegans
Authors: Johnsky Lily Renee , McGehee Annette 
Published: June 2021

An organism's nervous system processes decisions on how to respond to their environment. Studying how proteins of the nervous system are regulated enables us to identify how an organism responds to its environment and alters its behavior. In our lab, we study glutamate receptors, specifically GLR‐1. Glutamate is used for communication between neurons, and changes in glutamate receptors are believed to play an important role in learning and memory. The DAF‐7/TGF‐β signaling pathway regulates GLR‐1 in C. elegans and mutants in the DAF‐7/TGF‐β signaling pathway have increased GLR‐1. However, the mechanism through which GLR‐1 levels are increased in the daf‐7 mutant worms is unknown. My research addressed how GLR‐1 levels were increased in the daf‐7 worms. I explored the possibility that the transcription factor DMD‐10 is affected in the daf‐7 mutants, and then in turn affects GLR‐1. We tested this by measuring spontaneous reversals (a GLR‐1 dependent behavior) in both single mutant and double mutant worms. If DMD‐10 is affected in the daf‐7 mutants, we predict that the dmd‐10; daf‐7 double mutants would have fewer reversals than the daf‐7 mutant. The dmd‐10; daf‐7 double mutants did not have fewer spontaneous reversals than the single daf‐7 mutant, which suggests that the DAF‐7/TGF‐β signaling pathway does not use DMD‐10 to alter levels of GLR‐1. However, the dmd‐10 single mutant had fewer reversals than the wildtype, suggesting that DMD‐10 is required for normal reversals. The results of this experiment suggested that although DMD‐10 is not involved in DAF‐7/TGF‐β regulation of GLR‐1, it may have an independent effect on GLR‐1. My current research is aimed at determining if DMD‐10 is involved in the regulation of GLR‐1. I have generated a strain of dmd‐10 mutant worms that expresses GFP‐tagged GLR‐1 (GLR‐1::GFP). Imaging of GLR‐1::GFP in wild type and dmd‐10 mutant worms will allow us to test if DMD‐10 is involved in GLR‐1 regulation. This strain is being analyzed for total levels of GLR‐1::GFP using western blotting and will indicate if DMD‐10 is reducing total levels of GLR‐1. Synaptic levels of GLR‐1::GFP will be analyzed using fluorescence imaging that provides information on the location of GLR‐1 and whether it is localized in the cell body or synapses. Together, these results will indicate if DMD‐10 is affecting GLR‐1 and will give us insight into how regulation is occurring.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Expression of Piccolo Splice Isoforms During Cerebellar Development
Authors: Fenster Steven Daniel 
Published: June 2021

Piccolo is a multi‐domain protein with restricted expression to the presynaptic active zone, the site of synaptic vesicle fusion and neurotransmitter release. Alternative splicing of the Piccolo gene (PCLO) results in two primary splice isoforms: isoform‐1 (I‐1) with two C2 domains (C2A and C2B) and isoform‐2 (I‐2) with only a single C2A motif. In addition, an exon‐skipping event generates Piccolo isoforms that alter the C2A domain resulting in exclusion of a 27‐nucleotide exon located in the C2A domain (e.g. short variant). The result is a lack of 9 amino acids in the domain resulting in increased calcium and phospholipid binding affinity versus the longer C2A domain (e.g. long variant) that includes the 9‐peptide region. Consistent with the importance of the Ca2+‐binding property of C2 domain‐containing proteins in synaptic function, genome‐wide association studies have identified variations in or near the C2A domain of human PCLO that predispose individuals to affective disorders. The most compelling evidence for the importance of the C2A domain comes from recent identification of the genetic cause of pontocerebellar hypoplasia type III (PCH3): a neurological disorder that results in severe underdevelopment of the cerebellum. Analysis of genomic DNA from two affected individuals with PCH3 identified a homozygous nonsense variant in PCLO that introduces a STOP codon in the 3′ coding region including the C2A domain. The purpose of this study was to examine the temporal expression of Piccolo splice variants during critical periods of mouse cerebellar and cortical development. We performed real‐time quantitative RT‐PCR gene expression analysis of Piccolo splice isoforms (e.g. I‐1, I‐2, long and short) from RNA isolated from cerebellum at key stages of cerebellar granule cell differentiation (P0, P5, P15, adult) and also from the cortex at the same ages. Our analysis demonstrated that expression levels of the I‐1 and I‐2 splice variants were relatively consistent between cerebellum and cortex at all examined developmental time points. In contrast, we showed that during initial stages of cerebellar development (P0 and P5), the more calcium‐sensitive short variant had significantly increased expression levels relative to the long variant. Our results indicate that increased expression of the short variant may play a role in neuronal proliferation during initial cerebellar development when considering the augmented calcium permeability of immature neurons during this time period.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Suppression and inhibition of Acetylcholinesterase (AChE) Gene Expression and Adenosine Deaminase (ADA) respectively in Cadmium Treated Rats by curcumin administration
Authors: Fadaka Adewale Oluwaseun , Akinyemi Ayodele J. , Adewale Olusola Bolaji , Olayide Israel Robinson , Onikanni Amos , Olaoye Oyindamola Adeniyi , Okoh Patrick Osemeyekeh 
Published: June 2021

Curcumin, the main polyphenolic component of turmeric (Curcuma longa) rhizomes have been reported to exert anticholinesterase and cognitive enhancing potentials with limited information on their scientific basis and how they regulate gene expression.This study sought to evaluate the in vitro effect of curcumin on cerebral cortex acetylcholinesterase (AChE) activity, their mRNA gene expression level and adenosine deaminase (ADA) in cadmium (Cd)‐treated rats. Animals were divided into six groups (n=6): group 1 [(control) received saline/vehicle], group 2 [saline plus curcumin (25 mg/kg)], group 3 [saline plus curcumin (50 mg/kg)], group 4 [Cd (2.5 mg/kg) plus vehicle], group 5 [Cd (2.5 mg/kg) plus curcumin (25 mg/kg)] and group 6 [Cd (2.5 mg/kg) plus curcumin (50 mg/kg)] by oral gavage for 7 days. Acetylcholinesterase (AChE) and adenosine deaminase (ADA) activities were determined and AChE expression was analyzed out using real time polymerase chain reaction (RT‐qPCR).The results of this study revealed that ADA activity was increased in Cd‐poisoned rats, and curcumin co‐treatment reversed this effect comparable to the control. Also, it was observed that acute administration of Cd increased AChE activity and caused a significant (P<0.05) increase in AChE mRNA levels in whole cerebral cortex when compared to control group. However, co‐treatment with curcumin inhibited AChE activity and alters AChE mRNA levels when compared to Cd‐treated rats. Furthermore, Cd intoxication increased the level of lipid peroxidation in cerebral cortex with a concomitant decreased in functional sulfhydryl (−SH) group and nitric oxide (NO). However, the co‐treatment with curcumin at 25 and 50 mg/kg, respectively increased the non‐enzymatic antioxidant status and NO in cerebral cortex with a decreased in malondialdehyde (MDA) level.In conclusion, inhibition of AChE and ADA activities, suppression of AChE mRNA gene expression level in Cd exposed rats, as well as increased antioxidant status by curcumin in Cd‐induced memory dysfunction could suggest some possible mechanism of action for their cognitive enhancing properties, thus providing some biochemical and molecular evidence on the therapeutic effect of this turmeric‐derived compound in treating neurological disorders including Alzheimer's disease.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.