bluuebunny/crossref_metadata_2025_split huggingface dataset:
Degradation of Proteins Extracted from Teeth by Hydrogen Peroxide
Authors: Tadros Jonathan , Patel Feny , Keenan Kelly 
Published: June 2021

The active ingredient in whitening strips and professionally done procedures is hydrogen peroxide (H2O2) which damages proteins. Teeth were either untreated or treated, either one or three rounds, with over‐the‐counter whitening strips in artificial saliva, and dialyzed against 10% EDTA pH 9.0 for two weeks. The dialysis fluid contained the non‐collagen protein from the enamel and dentin layers of the teeth. The amount of protein was measured using a Lowry assay and the individual proteins were separated using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS PAGE) and stained with silver. The amount of protein extracted decreased with H2O2 treatments. The molecular weights (MW) for the proteins on the gel were correlated with the predicted MW for proteins known to be in the tooth and the impact of H2O2 was assessed. In order to demonstrate the effect of H2O2, pure proteins were pre incubated with either it or water for an hour and trypsin was added. Samples were collected, run on SDS PAGE and the impact on MW was assessed. The results show a loss of protein with a decrease in size when the proteins were treated with H2O2. These results suggest that if the active ingredient in whitening treatments can penetrate the enamel, it can damage the non‐collagen proteins in the teeth. This work was supported by Research and Professional Development Grant from Stockton University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neural Stem Cell Fatty Acid Beta‐Oxidation and Autism Spectrum Disease
Authors: Bankaitis Vytas A. , Xie Zhigang 
Published: June 2021

Inborn errors of metabolism occur with high incidence in human populations, and inborn deficiencies in fatty acid b‐oxidation (FAO) are clinically associated with neurological deficits such as speech weakness, motor delay, and autism. Using in vivo assays for analyzing neural stem cells (NSCs) in the developing mammalian forebrain, we find that neural stem cell (NSC)‐autonomous deficiencies in long‐chain FAO deplete neural stem cell (NSC) pools from mouse embryonic neocortex. Acute interference with activity of TMLHE (catalyzes the first step in carnitine biosynthesis), or with CPT1A (the rate‐limiting enzyme for carnitine‐dependent transport of long‐chain fatty acids into mitochondria), results in impaired NSC self‐renewal. Moreover, dorsal forebrain‐specific eviction of the Cpt1a gene resulted in reduced thickness of the murine neocortex at birth. Importantly, NSC deficits induced by TMLHE insufficiencies are rescued by exogenous carnitine in both cultured explants and by supplementation of the maternal diet. Taken together, the data argue for a direct role for FAO in controlling NSC self‐renewal vs differentiation in mammalian embryonic brain. The data further argue that deranged stem cell homeostasis is a significant mechanism for linking IEMs with human cognitive disorders such as autism. The implications of these findings for new strategies for management of autism risk will be discussed.Support or Funding InformationThis work was supported by the Robert A. Welch Foundation Award BE0017 and grant RO1GM112591 from the National Institute of HealthThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiolipin Modulation in the Mammalian Heart
Authors: Bick Diane , Holmes Michael , Sparagna Genevieve 
Published: June 2021

Cardiolipin (CL) is a phospholipid found in the inner mitochondrial membrane. It is essential for the optimal function of many of the protein components of the mitochondrial electron transport chain, metabolism and programmed cell death. CL has four fatty acyl chains. In the normal adult mammalian heart these fatty acyl chains are predominantly linoleate (18:2), making the dominant species tetralinoleoyl CL ((18:2)4CL). Research indicates that the composition of the CL acyl chains is altered in disease states associated with mitochondrial dysfunction like heart disease, ischemia and oxidative stress. The specificity of the CL side chain composition is not a result of specificity in CL biosynthesis. Specificity is determined by a remodeling process by which is either deacylation/reacylation or transacylated by enzymes using other phospholipids as acyl donors.We hypothesized that observed changes in cardiac cardiolipin levels may be the result of altered expression of enzymes in involved in the synthesis and/or remodeling of cardiolipin. Quantitative RT‐PCR analysis of several of these genes indicates a significant change in expression as a result of changes in diet and induced heart failure, conditions where changes in CL composition are documented. Supported by NIH HL072770 to DB
Akt mediates mitochondrial protection in cardiomyocytes through phosphorylation of mitochondrial hexokinase‐II
Authors: Miyamoto Shigeki , Murphy Anne N. , Brown Joan Heller 
Published: June 2021

Akt activation supports survival of cardiomyocytes against cell death through opening of the mitochondrial permeability transition pore (PT‐pore). Mitochondrial depolarization induced by H202 is prevented by activation of Akt with leukemia inhibitory factor (LIF). This protective effect is observed even when cardiomyocytes treated with LIF are permeabilized and mitochondrial depolarization is elicited by elevating Ca2+. Cell fractionation studies demonstrate that LIF treatment increases both total and phosphorylated Akt in the mitochondrial fraction and that mitochondrial Akt associates with hexokinase‐II (HK‐II). HK‐II contains consensus sequences for phosphorylation by Akt and LIF treatment induces PI3K‐ and Akt‐dependent HK‐II phosphorylation. Addition of recombinant active Akt to adult mouse heart mitochondria directly stimulates phosphorylation of HK‐II and concomitantly inhibits the ability of Ca2+ to induce cytochrome‐c release; this effect of Akt is prevented when HK‐II is dissociated from mitochondria by incubation with glucose 6‐phosphate or HK‐II dissociating peptide. Finally LIF stabilizes HK‐II association with mitochondria and requires mitochondrial HK‐II to mediate cardiomyocyte protection. We conclude that Akt protects against oxidant or Ca2+‐stimulated PT‐pore opening through a direct effect at the mitochondria mediated via phosphorylation of mitochondrial HK‐II.AHA Beginning Grant in Aide (Shigeki Miyamoto); NIH PPG 82521
Innate and adaptive immunity through autophagy
Authors: Munz Christian , Schmid Dorothee , Lee Monica , Gannage Monique , Paludan Casper 
Published: June 2021

Major histocompatibility complex (MHC) class II molecules present products of lysosomal proteolysis to CD4+ T cells. Although extracellular antigen uptake is considered to be the main source of MHC class II ligands, a few intracellular antigens, including the nuclear antigen 1 of the Epstein Barr Virus, have been described to gain access to MHC class II loading after macroautophagy. However, the general relevance and efficacy of this pathway is still debated. In addressing these questions, we could demonstrate constitutive autophagosome formation in MHC class II positive cells, including dendritic, B and epithelial cells. The autophagosomes continuously fused with multivesicular MHC class II‐loading compartments (MIICs). This pathway was of functional relevance, because targeting of the Influenza Matrix Protein 1 (MP1) to autophagosomes via fusion to the autophagosome‐associated protein Atg8/LC3 led to strongly enhanced MHC class II presentation to CD4+ T cell clones. The importance of this pathway is demonstrated by the fact that both DNA and RNA viruses inhibit the pathway, and might thereby escape immune control. We suggest that macroautophagy constitutively and efficiently delivers cytosolic proteins, like viral antigens, for MHC class II presentation and can be harnessed for improved helper T cell stimulation.This research is supported by the National Cancer Institute (R01 CA101741), the National Institute of Allergy and Infectious Disease (RFP‐NIH‐NIAID‐DAIDS‐BAA‐06‐19), the Beckman Foundation, the Sinsheimer Scholars Program and the Burroughs Wellcome Fund.
Neutrophil extracellular trap formation by nitric oxide donors: Involvement of nitric oxide synthase and myeloperoxidase derived free radicals
Authors: Patel Satyananda , Kumar Sachin , Saluja Rohit , Barthwal Manoj Kumar , Mitra Kalyan , Verma Anupam , Bajpai Virendra K , Dikshit Madhu 
Published: June 2021

Presence of neutrophil extracellular traps (NET) at the site of inflammation and infection is demonstrated in various pathological conditions. Neutrophils migration and augmentation in NO availability are mostly associated with inflammation and pathogen infections. So far role of NO has not been explored in NET formation. Present study reports for the first time that incubation of human neutrophils with NO donors, sodium nitroprusside (SNP) or S‐nitroso‐N‐acetyl‐penicillamine (SNAP) led to the NET release and free radical generation, which was blocked in the presence of free radical scavengers (ebselen, NAC) or pretreatment of PMNs with inhibitors of NOS reductase domain (7‐NI, DPI, imidazole) and myeloperoxidase (ABAH), while NOS oxygenase domain inhibitors had no effect. We thus propose that NO mediated NET release was dependent on the NO synthase (NOS) uncoupling and MPO derived free radicals, but independent of NADPH‐oxidase. Moreover, NO dependent free radical generation, NET release and phosphorylation of MEK/ERK, p38MAPK was blocked by inhibitors of MEK/ERK (U0126), p38MAPK (SB202190), PKC (Rottlerin) suggesting for the first time role of ERK and p38MAPK in NET formation. The results of the present study seem to have important bearing on the role of NO in inflammatory pathological conditions.The work was supported by CSIR and DBT India
Protective effects of polyclonal iTreg induced by IL‐2 and TGF‐β on a lupus‐like syndrome
Authors: Zheng Song Guo , Wang Juhua , Horwitz David A. 
Published: June 2021

We have previously reported that the adoptive transfer of antigen‐specific Tregs generated by allogeneic antigen stimulation with TGF‐β prevents the appearance of lupus syndrome induced by injection of DBA/2 mouse spleen cells into (DBA/2 × C57BL/6) F1 mice. Here we show that CD4+CD25‐ cells polyclonally activated ex‐vivo with IL‐2 and TGF‐β (iTreg) have similar beneficial effects in vivo. A single injection of 5 million iTregs prevented increased IgG production, decreased anti‐dsDNA autoantibodies, suppressed proteinuria and prolonged survival of these mice. The treatment with anti‐TGF‐β or ALK5 almost completely blocked the suppressive capacity of iTreg following injection in vivo. Moreover, iTreg suppress CD80 and CD86 expression of dendritic cells (DC). Thus, the protective effect of iTreg is dependent upon TGF‐β and these cells have inhibitory effects on antigen‐presenting DC. The possibility the transferred iTregs can induce T cells in the recipient mice to become iTreg is being explored. Since SLE, a systemic, generalized autoimmune disease, is characterized by impaired production of IL‐2 and TGF‐β, the use of the cytokines ex‐vivo to repair defects in the numbers and functional activity of CD4+CD25+Foxp3+ Treg cells in this disease may have therapeutic potential.Supported by grants from the Arthritis National Research Foundation, Clinical Research Feasibility Fund, James H. Zumberge Faculty Research Innovation Fund, the Arthritis Foundation, ExCell Therapeutics, Nora Eccles Treadwell Foundation, and BD Biosciences.
Altered signaling and split anergy in lupus T cells
Authors: Wadhone‐Gangalum Pallavi , Dubey Shweta , Wang James , Singh Ram Raj 
Published: June 2021

MRL‐Faslpr/lpr (MRL‐lpr) and MRL‐Fas+/+ (MRL+/+) mice develop autoimmune disease that mimics human SLE. We have used these mice to address the role of T cells in lupus. We found that freshly isolated CD4+ T cells from the spleen of MRL‐lpr display features of in vivo antigen priming and activation as compared to MHC‐matched healthy strains (B10.BR and C3H). Autoimmune T cells also produce more IFN‐γ than control T cells. Surprisingly, however, MRL T cells exhibit reduced calcium flux, IL‐2 production, and proliferation. Such anergic phenotype is seen upon stimulation with anti‐CD3, but not when stimulated with PMA/ionomycin. Such split responsiveness (activated, but anergic) is particularly more profound in a subset of T cells, i.e., B220+ T cells that expand in MRL mice. B220 is an isoform of CD45, a transmembrane phosphatase that plays critical role in cell signaling. As compared to control strains, MRL T cells exhibit reduced expression of CD45RB and CD45RC isoforms, which correlates with altered expression of TCR signaling molecules, with enhanced constitutive phosphorylation of ZAP‐70 and ERK1/2. However, no further increase or a highly accelerated kinetics in p‐ZAP70 and p‐ERK1/2 is seen in MRL T cells as compared to controls. There was also a decrease in phosphorylation of a key target of CD45, Lck (Y505). Using p‐ERK1/2 as readout for T cell activation, we found that activation in MRL‐lpr T cells was inhibited at a relatively lower concentration of CD45 inhibitor as compared to control T cells. Inhibition of CD45 normalized the basal level of activation in MRL‐lpr T cells. These data lead us to hypothesize that CD45‐mediated regulation of TCR signaling is impaired in lupus T cells.Research Support: Supported in part by NIH.
Intracerebral BCG infection impairs Th17 response and autoimmunity in the central nervous system
Authors: Lee JangEun , Reinke Emily K. , Sandor Matyas , Fabry Zsuzsa 
Published: June 2021

Infectious agents have been demonstrated to contribute to the development of helper T cell‐mediated autoimmune diseases including multiple sclerosis (MS). We induced an autoimmune disease that affects the central nervous system (CNS), experimental autoimmune encephalomyelitis (EAE), in mice intracerebrally infected with Mycobacterium bovis strain bacille‐Calmette‐Guérin (BCG) to study the role of in situ infection induced inflammation on the clinical outcome of disease. We show that clinical and pathological signs of EAE did not develop in mice with intracerebral BCG infection. In parallel with this, we found intact myelin sheaths and decreased cellular infiltration in the spinal cord of BCG‐infected animals as compared to non‐infected EAE controls. Intracellular cytokine analysis of IFN‐γ and IL‐17 demonstrated a substantial reduction in MOG‐specific responses in both cytokines in BCG‐infected mice. However, the IFN‐γ response of non‐MOG specific, activated T cells in the CNS of BCG infected mice was equivalent, while these cells showed substantially reduced IL‐17 responses. In addition, BCG‐induced suppression of Th17 responses was maintained in IFN‐γ deficient animals. These data suggest that BCG infection‐mediated modulation of autoimmunity in the CNS is independent of IFN‐γ and is regulated via the Th17 pathway. Identification of the mechanisms that block CNS autoimmunity in situ in the nervous tissue will add to our understanding of the role of bacterial infections in CNS autoimmunity and could lead to the development of new strategies in manipulating autoimmune disease.[This research is supported by the National Institute of Health, USA R01‐NS 37570 01 awarded to Dr. Z. Fabry.]
Transduced B‐cells mediate tolerance by activating regulatory T‐cells
Authors: Skupsky Jonathan , Scott David 
Published: June 2021

FoxP3 has emerged as an intracellular marker for mouse T cells that have a regulatory phenotype. To study the kinetics of regulatory T cell induction in response to treatment from tolerogenic B cells, we have crossed the FoxP3‐GFP knock‐in mouse to our mouse model for hemophilia; we have also crossed the FoxP3‐GFP mouse to a transgenic mouse with an ovalbumin specific TCR. We use the established gene‐therapy technique of transducing LPS activated B cells with an antigen‐IgG fusion to prevent or abrogate an ongoing immune response. Although it has already been shown that tolerance is dependent on CD4+CD25+ regulatory cells, herein, we demonstrate an increase in levels of antigen specific FoxP3+ cells that peaks within a week of treatment and wanes near baseline levels by two weeks. Additionally, we see an increased response of regulatory cells following challenge with antigen. We anticipate that a deeper understanding of the kinetics of FoxP3 induction will likely lead to insight into treatment modalities to maintain high levels of regulatory cells during the management of autoimmune diseases, transplant, and protein based therapies.(Supported by NIH grants R01 HL061883 and T32 HL07698)
C5 complement inhibition contributes to increased proliferative activity and antigen specific recall response in experimentally acquired myasthenia gravis (EAMG)
Authors: Soltys Jindrich , Kusner Linda L , Zhou Yuefang , Gong Bendi , Kaminski Henry J 
Published: June 2021

Background: Myasthenia gravis (MG) is an autoimmune disorder caused by the production of autoantibodies against the acethylcholine receptor (AChR). Antibodies bind to the AChR and activate the complement system leading to the destruction of neuromuscular junction (NMJ). Experimentally acquired MG (EAMG) rats were used to investigate the therapeutic role of C5 complement inhibitor, rEV576.Methods: Severity of disease was measured by grip strength and weight loss of the animal; serum cytokines and toxicity; cells proliferative activity and antigen specific recall responses were analyzed in vitro.Results: Treatment with C5 complement inhibitor for 10 days attenuated the outcome of EAMG. Administration of rEV576 resulted in reduction of serum toxicity and increased IFN‐γ production. Splenic cell suspensions isolated from EAMG + rEV576 had higher proliferation ability in the presence of specific antigen (AChR) or mitogen (Con A). ELISPOT recall responses to the specific AChR showed that the frequency of splenic T‐cells secreting IFN‐γ is increased in rEV576 treated EAMG rats.Conclusions: Inhibition of complement activity attenuated the outcome of EAMG. EAMG rats treated with rEV576 showed increased cell activity suggesting an effect of complement deficiency on cellular immunity. The data demonstrate that complement inhibition may have significant therapeutic value in human MG.
Increased activity of the mTOR pathway contributes to deficiencies of CD3ζ and Lck in T cells isolated from patients with systemic lupus erythematosus
Authors: Fernandez David Rafael , Bonilla Eduardo , Perl Andras 
Published: June 2021

We previously showed that the immunosuppressant macrolide rapamycin (Sirolimus) effectively controlled SLE disease activity and normalized the high baseline calcium as well as the increased T cell receptor (TCR) stimulation‐induced calcium fluxing in SLE T cells. Here, we document that mTOR activity is increased in SLE T cells and it is normalized by in vivo treatment with rapamycin, a specific inhibitor of mTOR kinase activity.We show increased phosphorylation of two targets of mTOR, ribosomal S6 kinase (S6K) and eukaryotic initiation factor 4E binding protein (4E‐BP), in T cells from SLE patients. Healthy controls and patients treated with rapamycin in vivo did not show phosphorylation of these proteins. Levels of mTOR and of its binding partners, raptor and rictor, were similar in control and SLE T cells.SLE T cells are deficient in the TCR‐associated proteins CD3ζ and Lck. It has been shown that correction of the CD3ζ deficiency by expression of exogenous CD3ζ in SLE T cells normalizes calcium homeostasis. Rapamycin treatment in vivo also normalizes calcium fluxing in SLE T cells, so we measured the levels of CD3ζ and Lck in T cells isolated from rapamycin‐treated patients. T cells obtained from patients treated with rapamycin in vivo show significant recovery of these signaling molecules versus SLE patients not receiving rapamycin.Rapamycin is an efficacious treatment for SLE and normalizes calcium levels in SLE T cells, likely by restoring normal levels of T cell receptor‐associated signaling molecules.This research was supported by the Alliance for Lupus Research and NIH grants AI‐061066, AI‐48079.
Development of Inducible Pattern Recognition Receptors and A Unified Dendritic Cell Enhancement Vector
Authors: Narayanan Priyadharshini , Spencer David M 
Published: June 2021

Recognition of pathogen‐associated molecular patterns (PAMPS) via pattern recognition receptors (PRRs) on the surface of dendritic cells (DCs), results in their activation and maturation. PRRs include cell surface Toll‐like receptors (TLRs) and intracellular receptors including RNA helicases and NOD‐like receptors. Stimulation with a TLR4 ligand, in conjunction with CD40 co‐stimulation, leads to potent activation, maturation and migration of human monocyte‐derived DCs. To circumvent drawbacks associated with the use and clinical availability of synthetic TLR4 ligands, we set out to develop a simplified, unified vector consisting of an adjuvant‐responsive domain, namely, a TIR domain from a TLR and a CD40 costimulatory receptor domain. This chimeric receptor will enable optimal DC activation in vitro and in vivo using the chemically induced dimerization (CID) system, thus replacing the panel of activation reagents, currently in use to activate DCs in vitro. Toward our goal, we have developed several inducible PRRs. We are now comparing the efficacy of iPRRs using reporter assays that monitor downstream activation of transcription factors such as NF‐κB, IRF3 and IRF7 in various cell lines. We observe that iRIG‐I and iNOD2 activate NF‐κB several fold, while iTLRs show weak induction and require additional optimization. In conclusion, iRIG‐I and iNOD2 are potent activators of NF‐κB making them ideal candidates for fusion with the CD40 domain to make the chimeric receptor. The chimeric receptor will be tested in mouse bone marrow‐derived dendritic cells and used to generate “enhanced DCs” tested for their efficacy in mouse tumor models.Funding: R01‐CA120411
Provision of tumor‐specific CD4+ T cells to enhance tumor immunity in a murine model of prostate cancer
Authors: Shafer‐Weaver Kimberly Ann , Malyguine Anatoli , Hurwitz Arthur A 
Published: June 2021

T cell tolerance to tumor antigens is a considerable challenge to cancer immunotherapy. Our laboratory studies T cell tolerance to tumor antigens using the TRansgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model in combination with adoptive transfer of TcR transgenic T cells with specificity for a TRAMP tumor antigen. We previously reported that adoptive transfer of CD8+ (TcR‐I) cells into TRAMP mice resulted in rapid tolerization of the cells. The objective of the current study was to test the ability CD4+ TcR‐II cells to prevent or reverse TcR‐I cell tolerance. Naïve TcR‐II cells transferred into TRAMP mice proliferate in the lymph nodes, are transiently activated, and subsequently traffic to the prostate where they become tolerant by 10 days after transfer. We next tested whether the transient activation of TcR‐II cells was sufficient to prevent TcR‐I cell tolerization. Co‐transfer of naïve TcR‐II cells with TcR‐I cells initially enhanced the frequency, activation, survival and function of TcR‐I cells but this effect was not durable, as TcR‐I cells were eventually tolerized by 3 weeks after co‐transfer with TcR‐II cells. Furthermore, our preliminary data suggest that TcR‐II cells transferred 10 days after TcR‐I cells were unable to rescue TcR‐I cells from tolerization. Taken together, these data demonstrate that the powerful immunosuppressive environment of the developing TRAMP tumor can exert its effect on T cells, irrespective of provision of T cell help. On‐going studies will test whether conditioning TcR‐II cells affects their ability to prevent or reverse TcR‐I tolerance. By studying the ability of CD4+ T cells to enhance anti‐tumor CTL activity, we hope to elucidate novel approaches to stimulate a more potent anti‐tumor immune response.Supported in part by NCI IRP and DOD PCRP.