bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Cannabinoid Receptor 2 Prevents Retinoblastoma Cell Migration
Authors: Vecchini‐Rodriguez Clara M. , Oquendo Yolymar , Yudowski Guillermo , Flores‐Otero Jacqueline 
Published: June 2021

Retinoblastoma (RB) is a rare cancer of the retina and it is the most common intraocular malignancy present in children less than 5 years of age. Current treatments cure over 95% of RB when it is restricted to the internal part of the eye, however less than 10% of children with extraocular RB survive. The endocannabinoid system has been found to be effective anti‐cancer agents in different cancer types, yet in RB the role of cannabinoid receptors or their ligands has not been elucidated. Our main objective is to determine the role of the cannabinoid 1 receptor (CB1R) and the cannabinoid 2 receptor (CB2R) in regulating RB progression with an emphasis in tumor invasion, using human RB cells and human tumors. Data from our laboratory demonstrate that in contrast to CB1R, the RNA expression of the CB2R is upregulated in RB cells and tumors. We hypothesize that the migration and invasion of highly aggressive RB cells (Y79) is modulated specifically by the CB2R. Our results show a significant upregulation in the protein expression levels of CB2Rs in RB cells and tumors when compared to fetal retina. To elucidate the migration rate, we performed wound healing assays using Y79 cells that were exposed to a synthetic agonist that activates both, CB1Rs and CB2Rs, CP55,940. Results demonstrate a time‐dependent effect in the cell's migration upon activation of the receptors. Our data suggests a functional role for CBR2 in modulating RB cell behavior. Ongoing studies using flow cytometry assay will determine the effect of CP55,940 and 2AG, a different endogenous ligand, in promoting RB cell cycle and viability. Future experiments will test the effects of CBR2 specific ligands in halting optic nerve invasion in an in vivo orthotopic RB xenograft model, hence preventing brain metastasis.Support or Funding InformationGrants: RCMI‐NIMHD 8G12MD007600 to JFO; NIH NIH‐NIDA R01 DA037924 to GY; NIGMS‐RISE program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Platelets and their Pharmacological Regulation on Cancer Cell Immune Checkpoint PD‐L1 Expression
Authors: Lesyk Gabriela Maria , Poitras Erika , Jurasz Paul 
Published: June 2021

BackgroundCancer cells have the ability to activate platelets, and platelets play an important role facilitating hematogenous metastasis. One mechanism by which platelets promote metastasis is their ability to shield cancer cells from natural killer cell‐mediated death. Although much is known of how platelets affect the innate immune response, our understanding of platelet function in modulating the adaptive immune system in cancer is limited. A major negative regulator of the adaptive immune response in cancer is the immune checkpoint transmembrane protein Programmed Death Ligand 1 (PD‐L1). PD‐L1 interacts with its receptor PD on T‐cells resulting in T‐cell anergy and/or apoptosis. This cancer cell survival mechanism is exploited in various types of cancer, including lung and kidney. Interestingly, platelets upon activation secrete a number of factors that have the potential to increase cancer cell PD‐L1 expression. Therefore, we hypothesized that cancer cell‐activated platelets increase cancer cell PD‐L1 expression and that common anti‐platelet drugs inhibit this platelet‐induced up‐regulation of PD‐L1.MethodsPlatelets were isolated from the blood of healthy human volunteers. Human A549 lung and 786‐O renal cancer cells were incubated with and without platelets for 24 hours and cancer cell PD‐L1 surface expression was measured by flow cytometry and mRNA by qPCR. In other experiments, platelet‐cancer cell incubations were performed in the presence of anti‐platelet drugs acetylsalicylic acid (ASA – 100 mM), Prasugrel active metabolite (PAM – 10 mM) or Integrelin (10 mM).ResultsPlatelets caused a significant increase in PD‐L1 surface expression by A549 (6.7±3.1% of A549 vs. 16.3±3.9% of A549 + platelets, P < 0.05) and 786‐0 cells (11.2±3.1% of 786‐O vs. 21.8±4,8% of 786‐O + platelets, P < 0.05). This increase in surface expression occurred as a result of a 3‐fold increase in PD‐L1 mRNA in A549 incubated with platelets. Platelets did not express PD‐L1 mRNA. Importantly, anti‐platelet drugs Prasugrel active metabolite (PAM) and Integrelin, but not acetylsalicylic acid, prevented the platelet‐dependent increase in PD‐L1 expression by A549.ConclusionsPlatelets up‐regulate the transcriptional expression of PD‐L1 by cancer cells, and Prasugrel and Integrelin can inhibit this up‐regulation. Further experiments are needed to determine whether platelet‐induced expression of PD‐L1 on cancer cells protects them from T‐cell induced death, and whether investigation of anti‐platelet drugs as adjuvant therapy in addition to immune checkpoint inhibitors is warranted.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 Antibiotics on Gut Microbiome: A Metagenomics Analysis of Microbial Shift and Antibiotic Resistance in Antibiotic Treated Mice
Authors: Xu Lei , Chockalingam Ashok , Stewart Sharron , Li Zhihua , Rouse Rodney 
Published: June 2021

There are long standing concerns about the use of oral antibiotics and their impact on the normal gut microbiota/microbiome and the emergence of antibiotic resistance. More recently, drug‐microbiota/microbiome interactions have been identified that have impacted drug efficacy and/or safety through altered drug metabolism re‐enforcing these concerns. This project characterizes the effect of different antibiotic treatment on the composition of the gut microbiota / microbiome and the prevalence of antibacterial resistance genes in the gut of mice treated for UTI. This knowledge provides a foundation for stratifying the potential of different mono‐ and potentially combination antibiotic therapies to negatively impact health through the microbiota/microbiome. Mice were experimentally infected by inoculation of E.coli into the urinary bladder and subsequently treated with one of three antibiotics (ampicillin, ciprofloxacin, fosfomycin) commonly used for UTI. For each antibiotic, fecal samples were collected after 1, 2, or 3 days of treatment to compare and contrast influence on the gut microbiota and microbiome composition. Standard NGS procedures were used with bio‐informatics to conduct metagenome analysis on fecal samples. Although individual animal variation was observed, control animals generally had similar findings for each experiment. For each antibiotic, analysis of aligned sequences revealed a post‐treatment reduction in number of bacterial species and a shift in the relative prevalence of the remaining species. By 24 hours after treatment initiation, fosfomycin demonstrated a larger and more profound reduction in species than either ciprofloxacin or ampicillin. Within the first 24 hours, ampicillin has minimal reduction in species while ciprofloxacin was intermediate to ampicillin and fosfomycin. Ciprofloxacin and ampicillin treatment led to detection of several species not identified in controls that in some cases became one of the most prominent species detected. Fosfomycin treatment resulted in a significant increase in prevalence of a known antibiotic resistance gene in the gut microbiome. These preliminary studies indicate that the spectrum and degree of impact on the microbiota species will vary by individual antibiotic but that in all antibiotic treatments as much as 70 to 80% of detectable species are greatly reduced or eliminated. The data also show that antibiotic treatment can increase the reservoir of antibiotic resistance in the microbiome.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
RNF20 regulates adipocyte differentiation through modulating PPARγ
Authors: JEON YONGGEUN , LEE JAEHO , KIM JAEBUM 
Published: June 2021

During adipogenesis, numerous adipogenic transcription factors need to be sequentially expressed and tightly modulated. Among these transcription factors, peroxisome proliferator‐activated receptor gamma (PPARγ) is the most important one during adipogenesis. PPARγ is a nuclear receptor and its activity is sophisticatedly regulated through the binding of coactivators and corepressors. However, the activation mechanisms of PPARγ during adipogenesis are poorly understood. In this study, we demonstrate that the E3 ubiquitin ligase ring finger protein 20 (RNF20) would be involved in adipocyte differentiation by regulating PPARγ. In RNF20 heterozygous‐null mice, the size of adipose tissue and adipocytes were reduced. Knockdown of RNF20 suppressed adipocyte differentiation of 3T3‐L1 cells. Conversely, overexpression of RNF20 promoted adipogenesis. We observed that RNF20 bound to nuclear corepressor (NCoR1) and increased the activity of PPARγ by repressing NCoR1. Taken together, these results suggest that RNF20 would act as a positive regulator for adipocyte differentiation by enhancing the activity of PPARγ.Support or Funding InformationThis work was supported by NRF of Korea Grant MSIP 2011‐0018312 and BK21This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Method validation of analytical method for determination of extracts of rosemary in various food matrix
Authors: Choi Seung‐Hyun , Jang Gill‐Woong , Choi Sun‐Il , Jung Tae‐Dong , Cho Bong‐Yeon , Han Xionggao , Sim Wan‐Sup , Kim Young‐Cheul , Lee Ok‐Hwan 
Published: June 2021

Extracts of rosemary is a food additive used as antioxidant for edible oil, dressing and sausage. In the EU and China, extracts of rosemary is authorized as food additive with maximum amounts for each food but unauthorized in Korea. Therefore, analytical method to detect extracts of rosemary in food is need. A sensitive simultaneous analytical method to identify rosmarinic acid, carnosol and carnosic acid residues in edible oil, dressing, and sausage was developed and validated using high‐performance liquid chromatography with photo diode array detector (HPLC‐PDA). Validation parameters; specifity, linearity, LOD, LOQ precision and accuracy were attained. The validated method has an analytical range from 1.56 μg/mL to 400 μg/mL in edible oil, dressing and sausage with an LOD and LOQ range from 0.22 to 1.73 μg/mL and 0.66 to 5.23 μg/mL, respectively, depending on the analytes. The recoveries of extracts of rosemary ranged from 70.6 to 116.5% with relative standard deviations between 0.2 and 3.8%. These results showed that the analytical method developed in this study in appropriate for determination of extracts of rosemary identification and can be used to maintain the safety of edible oil, dressing, sausage products containing extracts of rosemary residues.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using Conformational Penalties to Assess the Structure Dependence of RNA Cellular Function
Authors: Al‐Hashimi Hashim M. 
Published: June 2021

RNA is increasingly recognized as a key player in biology, a novel drug target, and a molecular tool for bioengineering applications and synthetic biology. The degree to which the cellular activity of regulatory RNAs depends on their three dimensional structure can vary from RNA to RNA and can be difficult to determine experimentally. Here, we describe a new approach for assessing the structure dependence of RNA cellular activity that relies on using mutations to introduce conformational equilibria that push the RNA toward non‐native conformations. By comparing the conformational equilibria of these mutants with their impact on cellular activity, we determine the structure‐dependence of cellular function for two highly conserved regulatory elements from HIV‐1.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protein Palmitoylation Contributes to Extracellular Vesicle Production and Effect on Vascular Barrier Dysfunction during Systemic Inflammation
Authors: Yang Xiaoyuan (Sherry) , Chatterjee Victor , Creasy Alexandria , Zheng Ethan , Yuan Sarah Y. 
Published: June 2021

Extracellular vesicles (EVs) are membrane‐encapsulated particles released from various types of cells, ranging from 30–1000 nm in diameter. EVs facilitate intercellular communication by transferring proteins and lipid among cells not in direct contact. Recent studies suggest the involvement of EVs in the progression of systemic inflammatory response syndrome (SIRS) associated with trauma or infection. However, there is limited information about EV production, cargo content, or pathological function in systemic inflammation. Protein palmitoylation, the covalent attachment of palmitic acid to cysteine residues, catalyzed by a family of palmitoyl acyltransferases containing a zinc finger aspartate‐histidine‐histidine‐cysteine (DHHC) motif. Palmitoylation has recently been recognized as a novel signaling mechanism in regulating endothelial barrier function and microvascular permeability. In this study, we sought to examine whether protein palmitoylation affects EV production or function in microvascular barrier in response to systemic inflammation. EVs were isolated from the blood of wild‐type (WT) and DHHC21 deficient (Zdhhc21dep/dep) mice with SIRS. Nanoparticle tracking analysis showed an increased plasma level of EVs in WT mice subjected to SIRS compared to sham‐treated, but this response was diminished in Zdhhc21dep/dep mice. Next, isolated EVs from SIRS mice were injected into healthy mice intravenously to assess their effects on microvascular permeability. Intravital microscopic imaging analysis revealed that SIRS EVs from WT mice caused a massive leakage of plasma albumin into extravascular space, whereas SIRS EVs from Zdhhc21dep/dep mice produced less leakage. Furthermore, we examined the cargo contents of EVs produced during SIRS and identified an array of pro‐inflammatory molecules, including C‐reactive protein, interleukins and tumor necrosis factor‐a. Of particular note, SIRS EVs contained a high level of histones, which has been implicated in endothelial barrier injury. Such an EV profile was not observed in EVs from Zdhhc21dep/dep mice. Taken together, these data suggest systemic inflammation stimulates the production of pro‐inflammatory EVs that are capable of inducing microvascular leakage. This response is greatly attenuated in Zdhhc21 functional deficiency, indicating the involvement of palmitoylation in EV production or function.Support or Funding InformationThis work was supported by the National Institutes of Health Grants GM097270, HL070752, and HL126646.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lysosomal Metabolic Cues in Orchestrating Cellular and Organism Homeostasis
Authors: Wang Meng 
Published: June 2021

Lysosomes are highly specialized cellular organelles with crucial functions in metabolic digestion and recycling. They carry a variety of hydrolases that function at low pH and generate specific metabolic products. Work in my laboratory is interested in understanding how these metabolic activities of lysosomes are linked to their signaling communication with other cellular organelles, such as mitochondria, endoplasmic reticulum (ER) and the nucleus. We discovered a lysosome‐to‐nucleus retrograde lipid signaling pathway in promoting healthy aging and metabolic fitness, and revealed its critical roles in the metabolic and signaling interaction between lysosomes and mitochondria. Most recently, we discovered a lysosome‐specific nucleotide hydrolase and its key functions in the control of ER stress responses through interacting with mTOR signaling. These studies highlight the signaling role of lysosomes in the active coordination of cellular homeostasis and in the improvement of the healthy survival of an organism.Support or Funding InformationThis work was supported by HHMI and National Institutes of Health.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Designing new protein structures and functions with the molecular modeling program Rosetta
Authors: Kuhlman Brian 
Published: June 2021

Proteins perform an amazingly diverse set of functions in all aspects of life. Critical to the function of many proteins are the highly specific three‐dimensional structures they adopt. For this reason, there is strong interest in learning how to rationally design proteins that adopt user‐defined structures. Over the last 20‐years there has been significant progress by us and others in the field of computational protein design as rotamer‐based sequence optimization protocols have been used to accurately design protein tertiary and quaternary structure. I will summarize how the molecular modeling program Rosetta is used to design new protein structures and describe how we have used this capability to create proteins that have important applications in research and medicine. I will highlight three protein design stories: (1) the engineering of light‐inducible proteins that can be used to recruit proteins to specific locations in the cell, (2) the use of protein interface design to create therapeutic bispecific antibodies, and (3) the de novo design of new protein structures from pieces of naturally occurring proteins.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lactobacillus rhamnosus GG‐mediated expression of leptin in the intestine induces cell proliferation
Authors: Darby Trevor M , Luo Liping , Naudin Crystal , Jones Rheinallt M 
Published: June 2021

Identifying the functional elements that mediate efficient gut epithelial development and homeostasis is essential for understanding intestinal health and disease. Some of the molecular events that function in optimal gut epithelial health are induced after the colonization of the intestine by symbiotic bacteria. Some lactobacilli, when in contact with the gut epithelium, induce the activation of signaling pathways that function in cell proliferation, cell migration, epithelial wound sealing and cytoprotection. These processes involve the generation of non‐toxic physiological levels of ROS by NADPH oxidase (Nox1) and ensuing activation of the Nrf2 pathway signaling. However, the downstream functional elements that respond to Nox1‐generated ROS and mediate these events are yet to be fully described. Here, we show that ingestion of Lactobacillus rhamnosus GG induces elevated levels of Leptin in the gut epithelia, which as well as functioning in the context of metabolism, also has pleiotropic activity as a chemokine that induces cell proliferation. Using gut epithelial specific knockout mice, we show that L. rhamnosus GG –induced elevated levels of Leptin is dependent on a functional Nox1 in the colonic epithelium, and that L. rhamnosus GG –induced cell proliferation is dependent, on both Nox1 and on Leptin. We also show that L. rhamnosus GG induces JAK‐STAT signaling pathway in the gut in a Nox1 and Leptin dependent manner. Our data suggest that L. rhamnosus GG elevates Leptin levels in the gut epithelium, and that Leptin then acts as a chemokine promoting cell proliferation through the upregulation of JAK‐STAT pathway signaling. These results demonstrate a novel role for Leptin in the response to colonization by lactobacilli, where Leptin is involved in the transduction of signals from symbiotic bacteria to sub epithelial compartments, where it modulates intestinal development and homeostasis.Support or Funding InformationNIH Grants R01DK098391This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Mechanisms of Cue‐Induced Translation Regulation
Authors: Filbin‐Wong Megan E. , Gonen Tamir , Kieft Jeffrey S. 
Published: June 2021

Precise tuning of local gene expression is essential for proper neural development. In particular, environmental cues spatially and temporally direct neuron growth by stimulating or repressing protein synthesis. The integral membrane protein, deleted in colorectal cancer, or DCC, detects the environmental growth cue netrin‐1, and transduces the growth signal to a variety of intracellular responses, including activating protein synthesis. DCC is primed to function in this way; in the absence of netrin‐1, translation machinery is tethered to its C‐terminal tail (C‐tail) and silenced. Once netrin‐1 binds DCC, the receptor dimerizes, translation machinery is released, and protein synthesis commences. How DCC's C‐tail binds to the protein synthesis machinery and prevents translation in the absence of netrin‐1 is unknown. The C‐tail is a 35 kDa protein that is composed of three domains (P1–3) and is predicted 91% unstructured. Dimerization is facilitated by the P3 domain, while the P1 domain binds to the 60S subunit protein, ribosomal protein L5 (rpL5). Titrating the full‐length C‐tail into translation‐competent lysates, we found that the tail specifically inhibits eukaryotic translation, and inhibits protein synthesis of messages that use both canonical and non‐canonical translation initiation mechanisms. Hence, we propose that the C‐tail functions primarily through binding the ribosome, not eukaryotic initiation factors. We have evidence from cosedimentation and in vitro binding studies to show that the full‐length tail binds to the small and large subunits, as well as the 80S ribosome in the absence of mRNA. Binding and luciferase reporter assays show that the P3 dimerization domain is dispensable for translation inhibition, and the P1 domain is necessary but not sufficient to bind the ribosome and decrease translation. Thus the P2 domain and/or linker sequence(s) are also important for function. Collectively, our results suggest that DCC's C‐tail inhibits translation by effectively preventing ribosomes from binding to messages. We hypothesize the unstructured C‐tail precludes mRNA binding by interacting with the decoding groove of the ribosome until receptor dimerization releases 80S ribosomes. Thus chemoattraction via netrin‐induced DCC receptor dimerization might lead to a global (not message‐specific) increase in localized protein synthesis at axonal growth cones.Support or Funding InformationMSU Denver Provost Seed GrantMSU Denver College of Letters, Arts and Sciences Dean's Seed GrantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.