bluuebunny/crossref_metadata_2025_split huggingface dataset:
Engineering of Enzymes to Improve Lignin Breakdown for Use in Fuel Ethanol Production
Authors: Greene Russell D. , Hall Bonnie L. 
Published: June 2021

Lignin is the second most abundant biopolymer on Earth, behind cellulose. Lignin contains aromatic rings similar to those in fossil fuels, and breaking down lignin into smaller polymers is the first step in using it as a source of fuel. Once lignin is broken down, additional enzymes can then hydrolyze the cellulose and hemicelluloses, so that sugar fermentation and production of ethanol can occur. Lignin is difficult to break down, although some fungi can slowly degrade it using lignin peroxidase and/or laccase enzymes. Laccases break down the complex structure of lignin, while lignin peroxidases oxidize the aromatic rings. These are slow processes and are not practical for mass‐producing fuels from lignin. We are engineering these enzymes with the goal of improving the efficiency of lignin degradation. We have begun by studying BglB, an enzyme involved in the hydrolysis of cellulose, as a proof of concept. We have successfully expressed, purified, and assayed the activity of the BglB protein. We have also used FoldIt software to identify mutations for modifying the activity of the BglB protein. One specific series of mutants characterized had the amino acid substitutions W402S, W412F, E409Q, and W410H. We have characterized the activity of these mutants, and found W412F had similar activity as the wild type, while the activities of W402S, E409Q, and W410H were lower than wild type. We are now ready to expand these engineering methods to lignin peroxidase and laccase enzymes, to generate more efficient enzymes for the production of fuel ethanol from lignin.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Computational Insights into the Structure and Epitope locations for Two Idiopathic Membranous Nephropathy Antigens: Phospholipase A2 Receptor and the Thrombospondin Type‐1 Containing Domain 7A and Design of Antigen Binding Proteins
Authors: Stoddard Shana 
Published: June 2021

The use of non‐specific treatment options in autoimmune therapy today unfortunately causes patients to be prone to other infections. This non‐specific therapy suppresses the entire patient's immunes system also making it harder to fight of basic infections. Thus therapies need to be developed which can accurately target only the aberrant portion of the immune system that contributes to the autoimmune disorder (AD) specific to the individual. In the AD idiopathic membranous nephropathy (IMN) two antigens have been determined to produce an immune response. The work here focuses was performed to aid in the design of antigen specific binding proteins (ABP) as a targeted means to treat AD, and specifically IMN. In this work a homology model of the thrombospondin type‐1 containing domain 7A (THSD7A), was successfully developed, and epitope regions were predicted using Epitopia. In silico mutagenesis was utilized to develop ABP for both THSD7A and phospholipase A2 receptor (PLA2R). Using Rosetta protein‐protein docking simulations evaluate for maximum binding efficiency against the ricin and the CTLD2 epitopes present on theTHSD7A and PLA2R antigens. Our results show that THSD7A has 22 domains and 12 domains have potential to contain epitope sites. It was determined that there were two possible epitope locations in CTLD1 domain and three epitope locations in CTLD7 domain of the PLA2R antigen. Our results show improved binding scores and selectivity of ABP against the PLA2R. Initial binding energy, measured in rosetta energy unit (REU), were −2.863 and −2.633 for the SH13 template and ySMB‐1 ABP templates respectively against the PLA2R antigen. The M005 mutation designed of the SH13 improved binding scores to −5.32 REU. Mutations to the ySMB‐1 template were shown to be able to enhance scores to −6.907 REU. Molecular interactions leading to enhanced potency have also being evaluated. ABP candidates for the THSD7A antigen have also been designed. A total of fifteen ABP candidates have been selected for expression which was performed in E. coli followed by subsequent purification using affinity chromatography for experimental verification of computational results is using ELISA. This work has implications for new antigen specific approach to AD therapy.Support or Funding InformationFaculty Development Endowment Grant, Rhodes CollegeThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cytotoxic Mechanism of Long‐chain Lipids Extracted from Mexican Native Avocado Seed (Persea americana var. drymifolia) on Colon Cancer Cells
Authors: Lara‐Márquez Mónica , Spagnuolo Paul A. , Salgado‐Garciglia Rafael , Ochoa‐Zarzosa Alejandra , López‐Meza Joel Edmundo 
Published: June 2021

Cancer represents a worldwide problem, in 2012 about 8.2 million deaths were reported. Colorectal cancer (CRC) is the third most common type of cancer and the fourth leading cause of death in the world, accounting for about 700,000 deaths a year. Chemotherapy currently used in patients with CRC is associated with severe side effects. The use of natural compound combinations extends the possibilities of therapeutic targets. Long chain lipid molecules, such as long chain fatty acids, aliphatic acetogenins and polyhydroxylated fatty alcohols independently have been shown to have anticancer effects and have been proposed as chemo‐therapeutic adjuvants. The main objective of this work was to determine the cytotoxic effect of an extract rich in long chain lipids from Mexican native avocado seed (Persea americana var. drymifolia) on colon cancer cells. Colon adenocarcinoma cell line Caco‐2 was used and viability was measured by trypan blue exclusion assays. Changes in mitochondrial membrane potential, cytokine expression, intracellular calcium levels, cell cycle analysis and ROS production were determined by flow cytometry. The induction of apoptosis was analyzed by flow cytometry; spectrometry and fatty acid oxidation (FAO) rate determination was made using high resolution respirometry. The results showed that the long chain lipids from avocado seed induced apoptosis and the activation of the caspases 8 and 9, loss of the membrane mitochondrial potential (68%), the inhibition of FAO (45%) and the increase in ROS production (30%); however, the intracellular calcium levels were not modified. In addition, cell cycle arrest in the G1/G0 phase was induced. Also, a modulation in cytokine expression (IL‐6, IL‐8, IL‐10 and IL‐1β) was observed. In conclusion, long‐chain lipids from Mexican native avocado seed are cytotoxic towards Caco‐2 cells, inhibiting the FAO which increases ROS production and thus favoring the loss of membrane mitochondrial potential that allows the induction of caspase‐dependent apoptosis. In addition, these molecules induce arrest of the cell cycle and the modulation of cytokines related to the inflammatory response.Support or Funding InformationCIC‐UMSNH 14.5This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluating the Inhibition of Palmitoylation and NHE1 on Cell Proliferation and Migration
Authors: Hanowski Stephanie A. , Provost Joseph J. , Wallert Mark 
Published: June 2021

The Na+‐ H+ Exchanger Isoform I (NHE1) is a key regulator of cell proliferation and migration in a range of cell types from both healthy and diseased tissues. Activation of NHE1 is an initial step in the development of the transformed phenotypes of cancer cells and therefore a key driver for tumor development and cancer progression. NHE1 also contributes to two of the hallmarks of cancer, the activation of invasion and metastasis and sustained proliferative growth. We have identified for the first time, that NHE1 undergoes reversible palmitoylation, the addition of a palmitic acid residue to the cytoplasmic regulatory domain of NHE1. We hypothesize that this process regulates NHE1 activation and will play a role in the control of cell proliferation and migration. This suggests that targeting the potential palmitoylation sites on NHE1 may provide a therapeutic target for cancer treatment. Our work characterizes the role of the palmitoylation of NHE1 in cell growth and migration. Using the palmitoylation inhibitor, 2‐bromopalmitate (2BP), we evaluated cell growth and migration in the presence and absence of palmitoylation. In proliferation assays, the rate of cell growth increased approximately 5‐fold when the serum present in the growth media was increased from 0.5% to 10% and approximately 2‐fold when 50 μM lysophosphatidic acid was added to the growth media. Under both conditions, the addition of 15 μM 2BP or 10 μM cariporide dramatically reduced or eliminated the increased proliferation. Cell migration was evaluated using Electric Cell‐substrate Impedance Sensing (ECIS). Once again, the rate of cell migration was reduced by 50% or more in the presence of 15 μM 2BP or 10 μM cariporide. These data demonstrated that inhibiting the palmitoylation of NHE1 decreases both cell proliferation and the rate of cell migration.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 Effect of Resveratrol in the Proliferation and Autophagy of Breast Cancer Cell Lines
Authors: Casillas Maryangie Martinez , Forti Kevin Munoz , Rivera Jose Robles , Torres Geraline Trossi , Martinez Edu Suarez , Rivera Abigail Ruiz 
Published: June 2021

Breast cancer (BC) is one of the most universal malignant tumors of worldwide in women and it is a significant public health problem in terms of both morbidity and mortality. BC is the second leading cause of death among American women. It is estimated 250,000 new cases of invasive BC will be diagnosed in women in 2017. For instance, there still existing the need for developing or identify new effective compounds to fight this disease. For example resveratrol, which is a plant derived polyphenol with anti‐estrogenic and antioxidant properties. In‐vitro studies indicated resveratrol has anticancer properties such as cell cycle arrest, apoptosis induction, prevention of DNA damage, and angiogenesis inhibition. However, scientific literature studying the effect of resveratrol on DNA synthesis and in autophagy is limited. For this reason we studied the effect of resveratrol in the proliferation of three breast cell lines CCD‐1074 (non‐malignant cells), CCD‐1097 (transitional cells), and MCF‐7 (malignant non‐invasive cells); and autophagy induction in the MCF‐7 cell line. We hypothesized that resveratrol will decrease cell proliferation and increase autophagy in BC cells when compared to control treatments. We studied the proliferation effect in cells treated with resveratrol at 25, 50, 100, 200 μM during 24 hours post treatment. We used Bromodeoxyuridine (BrdU) and the detection of translocation of LC3 via flow cytometry to study proliferation and autophagy, respectively. Our results showed a significant decrease in cell proliferation at resveratrol concentrations of 100 and 200 μM across all cell lines when compared to the vehicle control (p<0.05). Importantly, at 50 μM resveratrol affected only transitional and malignant cell lines without affecting our non‐malignant cell line. Induction of autophagy was significantly higher when MCF‐7 cells were treated with 100 μM and 200 μM compared to the vehicle control (p <0.05). However, we are currently in the process of evaluating the effects of resveratrol in other malignant and non‐malignant cell lines. We consider that the 50 μM concentration as a possible therapeutic window selectively to inhibit cell proliferation and induce autophagy.Support or Funding InformationDo not applyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Calcitriol increases leptin expression in neuronal cells ‐ Implications for Alzheimer's Disease
Authors: Marwarha Gurdeep , Ghribi Othman 
Published: June 2021

Alzheimer's disease (AD) is a devastating progressive neurodegenerative disorder that is neuropathologically characterized by the intracellular accumulation of hyperphosphorylated protein tau as neurofibrillary tangles and the extracellular deposition of amyloid‐beta (Aβ) peptide as amyloid plaques. However, the most accurate pathological hallmark and clinical correlate of cognitive impairment as well as deficits in learning and memory is synaptic loss. Leptin is a pleiotropic adipocytokine known to be expressed endogenously in the mammalian brain that plays an indispensable role in learning and memory. Leptin is a bona‐fide neurotrophic cytokine and a growth factor that is integral in the regulation of a multitude of physiological and biochemical neuronal processes. Emerging evidence has implicated that the dysregulation and attenuation of leptin expression in the brain may lead to the initiation and fostering of deleterious neurodegenerative cascades that could culminate in cognitive dysfunction, learning and memory deficits, as well as AD‐like pathological hallmarks. More importantly, leptin decreases Aβ plaques and hyperphosphorylation of protein tau, two pathological hallmarks of AD, and rescues cognitive impairment in several rodent models of AD. A multitude of epidemiological studies, prospective and cross‐sectional, have implicated that lower plasma vitamin D3 levels are associated with a higher risk for developing AD. Contemporary studies have shown that vitamin D3 can regulate leptin expression in the adipose tissue and modulate peripheral circulating plasma levels of leptin. However, no studies have yet examined the effects of Vitamin D3 on leptin expression in the brain and neuronal cells. In this study, we determined the effects of calcitriol (1,25 dihydroxyvitamin D3) on leptin expression in cultured SH‐SY5Y human neuroblastoma cells. Herein, we show that calcitriol (1,25 dihydroxyvitamin D3) increases leptin expression levels by Vitamin D Receptor (VDR) – dependent and – independent mechanisms. Further elucidation and delineation of the underlying mechanisms unveiled the indispensable involvement of the CCAAT/Enhancer Binding Protein alpha (C/EBPα) in the 1,25 dihydroxyvitamin D3 (calcitriol) – induced increase in leptin expression. Our study is the first to demonstrate that 1,25 dihydroxyvitamin D3 (calcitriol) modulates leptin expression in neuronal cells and delineate the signaling cascade. Our study highlights and characterizes a novel salutary role of Vitamin D3 in neuronal cells as it increases the expression of leptin, a pleiotropic cytokine in the brain known to thwart neurodegenerative cascades and augment neuronal survival.Support or Funding InformationThis work was supported by a Grant from the NIH (R01AG045264) to Dr. Othman Ghribi.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
17β‐Hydroxysteroid dehydrogenases and neurosteroid metabolism in the central nervous system
Authors: yang song‐yu , He Xue‐Ying , Dobkin Carl 
Published: June 2021

17β‐Hydroxysteroid dehydrogenases are indispensable for neurosteroidogenesis. Neurosteroids are synthesized de novo in neurons and glia from cholesterol transported into mitochondria, or by conversion from proneurosteroids, e. g. dehydroepiandrosterone (DHEA) and pregnenolone, through the same metabolic pathway as de novo neurosteroidogenesis. Proneurosteroids are generated from endocrine glands—the adrenal cortex, ovary, and testis—and then transported into neurons and glia via the blood circulation. Non‐hormonal neurosteroids bind to cell surface receptors, e.g., GABAA or NMDA receptors and elicit antidepressant, anxiolytic, anticonvulsant and anesthetic effects by regulating neuroexcitabilty. Both non‐hormonal and hormonal neurosteroids play important roles in brain development including neuroprotection, neurogenesis and neural plasticity. They are also a critical element in cognitive and memory functions. Mitochondrial 17β‐HSD10, encoded by the HSD17B10 gene mapping to Xp11.2, is found in various brain regions, and is essential for the maintenance of neurosteroid homeostasis. Mutations identified in this gene result in two distinct brain diseases, namely HSD10 deficiency and MRXS10. Their clinical presentations and pathogenetic mechanisms are quite distinct, thus, it is not appropriate to combine them as a single “HSD10 disease”. Since elevated levels of 17β‐HSD10 is found in brains of Alzheimer's disease patients and AD mouse model, and this fascinating protein is responsible for the mitochondrial metabolism of neurosteroids such as 5α‐androstane‐3α,17β‐diol and 17β‐estradiol, analysis of the impact of 17β‐HSD10 and its inhibitor, “β‐amyloid peptide (Aβ),” on the metabolism of neuroactive steroids offers a new approach to AD pathogenesis.Support or Funding InformationNYS Office for People With Developmental DisabilitiesThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mast Cell Proteases Activate Glia‐Neurons and Release Interleukin‐33 by Activating MAPKs
Authors: Duraisamy Kempuraj , Selvakumar Govindhasamy Pushpavathi , Thangavel Ramasamy , Ahmed Mohammad Ejaz , Raikwar Sudhanshu P. , Zaheer Smita , Iyer Shankar S. , Zaheer Asgar 
Published: June 2021

Parkinson's disease (PD) is characterized by inflammation‐dependent dopaminergic neurodegeneration in the substantia nigra in the brain. Inflammatory mediators released from activated microglia, astrocytes, neurons, T‐cells and mast cells mediate neuroinflammation and neurodegeneration in the brain. 1‐methyl‐4‐phenylpyridinium (MPP+), a toxic metabolite of the parkinsonian neurotoxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP)‐induces the release of inflammatory mediators from glial cells, neurons and inflammatory cells including mast cells. Mast cells play an important role in neuroinflammation and neurodegeneration. Mast cell proteases are implicated in neuroinflammation. However, the exact mechanism involved in this process is not yet clearly understood. In the present study, mouse bone marrow‐derived mast cells (BMMCs) and mouse fetal brain‐derived glia/neurons and astrocytes were incubated with MPP+, mouse mast cell protease‐6 (MMCP‐6) and MMCP‐7 in vitro. Inflammatory cytokine interleukin‐33 (IL‐33) released from these cells was quantitated by enzyme‐linked immunosorbent assay. MMCP‐6 or MMCP‐7 stimulates astrocyte and glia/neurons to release significantly more IL‐33 as compared with untreated control cells. MPP+ also induced IL‐33 release from mouse mast cells and glia‐neurons. Our results show that mast cell proteases and MPP+ activate mitogen‐activated protein kinases (MAPKs) p38 and extracellular signal–regulated kinases1/2 (ERK1/2) and release IL‐33 from these cells. We suggest that mast cell interaction with glial cells and neurons during neuroinflammation can be explored as a new therapeutic target for PD and other neuroinflammatory conditions.Support or Funding InformationThis work was supported by Veteran Affairs Merit Award I01BX002477 and National Institutes of Health Grants AG048205.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 β‐cell Sensitivity to Kisspeptin via Linkage to GLP‐1
Authors: Aragaki Adam K. , Weber Craig , Vagner Josef , Lynch Ronald 
Published: June 2021

Glucagon Like Peptide‐1 (GLP‐1) and Kisspeptin (KP) receptors are co‐expressed on pancreatic β‐cells, and activation of both receptors potentiate glucose stimulated insulin secretion (GSIS), although they appear to work via different downstream signaling pathways. Unlike GLP‐1, KP has a low affinity for receptor activation (EC50 20 nM vs >1 uM) making it difficult to use as a drug without significant side effects. We developed a bivalent ligand composed of GLP‐17–36 and KP‐10 binding domains linked together, to evaluate if linking KP to a high affinity ligand would promote its activity at low concentrations by presumably increasing its local membrane concentration after GLP‐1 binding. This feature may also provide some specificity for targeting to β‐cells which express both receptors. The effects of KP‐10, GLP‐1 and the KP‐10/GLP‐1 bivalent analog on insulin secretion from the INS 832/3 cell line were investigated. KP‐10 by itself had no effect on insulin secretion in the absence of glucose, but more than doubled GSIS (basal, 0.23 +/− 0.07; GSIS 0.60 +/− 0.02 ng/min) with an EC50 of ~25 μM. At saturating doses, KP‐10 (100 μm) and GLP‐1 (100 nM) had similar effects on GSIS, and together as monomers had an additive effect (0.79 +/− 0.07 (ng/min) consistent with activation of independent downstream signaling pathways. The dimer also did not activate secretion in the absence of glucose, but doubled GSIS much like the individual monomers (0.64 +/− 0.01 ng/min); that is, an additive response was not obvious. This finding suggests that either the KP‐10 moiety is not binding or is not active in the bivalent ligand, or that GLP‐1 activity is diminished in the bivalent construct such that the combined effect of GLP‐1/KP‐10 is diminished when compared to the additive effect or the monomers. We have evidence from previous studies that binding of the GLP‐1 moiety within a bivalent construct is decreased relative to monomeric GLP‐11, suggesting an additive response may be present. Currently, we are determining the ability of GLP‐1/KP‐10 to initiate downstream signaling from both ligands to resolve the question: is KP active in the GLP‐1/KP at concentrations where monomeric KP‐10 is not.Support or Funding InformationFunded by the American Diabetes Association and Juvenile Diabetes Research FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.