bluuebunny/crossref_metadata_2025_split huggingface dataset:
NOCICEPTIVE STIMULUS IN THE NEONATAL PERIOD AFFECTS SATELLITE GLIAL CELLS MORPHOLOGY IN ADULT LIFE, WITH DIFFERENCES BETWEEN MALE AND FEMALE RATS
Authors: Simoes Ana Leda Bertoncini , Silva Greice Anne Rodrigues , Giorgetto Carolina , Carmo Elisabete Cassia , Fazan Valeria Paula Sassoli 
Published: June 2021

Satellite glial cells (SGCs) are found in the dorsal root ganglia (DRG) surrounding completely and individually sensory neurons, forming a thin sheath cells with regulatory function of the neuronal microenvironment. During the nociceptive stimulus the SGCs are activated and express glial fibrillary acidic protein (GFAP). Sustained nociceptive stimulus may cause morphological changes in SGCs influencing the mechanisms of pain chronification that may differ between genders. Understanding the plasticity in peripheral pain pathways considering gender differences is crucial, particularly when noxious stimuli are applied to neonates, when the nervous system is still under rapid and intense remodeling. We investigated the density (number/area) of neurons in the DRG that were enveloped by GFAP immunoreactive SGCs. Male (N=6 per group) and female (N=6 per group) Wistar rats, 15 and 180 days old after neonatal nociceptive stimulation [1] were used. Right and left L5 DRG were removed, frozen and 8 μm thick criossections were incubated in rabbit polyclonal anti‐GFAP antibody (1:2.000; 24hr). Immunostaining protocol was used as described [2]. The DRG area was measured with the aid of computer software. Neurons enveloped by a ring of GFAP‐labeled SGCs were counted, their density (number/area) was calculated and data were compared between right and left sides, 15 and 180 days of age and genders. Differences were considered significant if p<0.05. There was an increased density of neurons enveloped by a ring of GFAP‐labeled SGCs on both genders, 15 days after the nociceptive stimulus, compared to controls, with females presenting a larger density of these neurons compared to males at this age, on the stimulated side. At age of 180 days, the difference between pain and control groups persisted only in females. The comparison between genders showed that males present a smaller density of neurons enveloped by a ring of GFAP‐labeled SGCs compared to females on both ages with statistical significance only at 15 days old groups. There is an extensive literature clearly pointing to the fact that men and women are different in their responses to pain. Women present increased sensitivity to pain and differ in pain intervention responsivity. Despite that sociocultural and psychological factors certainly play a role in these gender differences, evidence from animals studies reveal important, qualitative and quantitative differences at low levels of the neuroaxis. Our results add extra evidence that multiple biological factors and mechanisms underlie gender differences in sensitivity and responses to pain.Support or Funding InformationFinancial support: FAPESP, CNPq, CAPES and FAEPAThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Student Perceptions and Experiences with Team‐Based Learning (TBL) in a Graduate Histology Course
Authors: Azim Homaira M. , Condon Keith W. , Brokaw James J. 
Published: June 2021

Contrary to the traditional didactic ways of science teaching, Team‐Based Learning (TBL) emphasizes active learning, application of knowledge, and team collaboration in the classroom. All students enrolled in a fall semester 2017 graduate histology course were invited to participate in this study. Twelve volunteer participants were asked about their opinions on TBL as well as their perceptual and experiential definition of human learning. Data collection methods included questionnaire, focus group, and direct observation. Student perceptions of and experiences with TBL were examined and linked to the literature on Tuckman's stages of group formation in order to explain some of the group dynamics in the field. Also, student perceptions on the concept of how learning occurs in general and how TBL may help in the process were examined in the context of critical constructivist theory of learning. Qualitative data analyses were performed on data across the three data sets. In general, the students perceived TBL to be a valuable learning experience. The five overarching themes identified as the requirements for a successful TBL performance on the student part were: time management, teamwork, self‐directed learning, intrinsic motivation, and learning not to resist active learning. Key significance of the TBL model were found to be in: active learning, teamwork, higher order application of knowledge, and providing formative assessment opportunities. Students' suggestions on what a good science teacher should look like and how they may avoid authoritarian exercises of power in day‐to‐day classroom activities are also documented and discussed.Support or Funding InformationNone.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Single‐cell Resolution of Developing Tissues and Cell Identity Programming
Authors: Wold Barbara J. 
Published: June 2021

We are using multiple single‐cell methods and platforms to deconvolve embryo limb histogenesis, tracing cell type specification and differentiation of major resident and immigrating cell lineages. We also experimentally reprogram the identity networks of closely and distantly related starting cell types, combining and comparing results from RNA‐seq and seq‐FISH data. From the resulting data, we assemble cell comprehensive identity networks. We find substantial differences in expression‐signature saliencies in developing embryo systems that have implications for future single‐cell data uses and for the design of larger scale sc‐RNA atlases. Our embryo data are part of the ENCODE consortium project, and integration of the single cell RNA‐seq data with the companion ENCODE whole‐tissue atlas, including epigenetic data, are discussed.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multimodal single mouse and human cell ‘Omics: Is variability distinct across cellular modalities?
Authors: Eberwine James H. 
Published: June 2021

With the advent of single cell analyses many unanticipated cell biological phenomena have been discovered including, new classes of RNA, identification of RNA targeting sequences and perhaps most surprisingly the discovery of cell‐to‐cell variation in transcriptome abundances even in presumptively identical cells. There is large‐scale single cell RNA variability for different cell types that can't be explained as simple molecular or technical noise. These data have led to the hypothesis that there is a many‐to‐one relationship between transcriptome states and a cell's phenotype. In this relationship the functional molecular ratios of the RNA are determined by the cell systems' stoichiometric constraints, which underdetermine the transcriptome state. Because a broad set of multi‐genic combinations support a particular phenotype, changes in the transcriptome state do not necessarily lead to changes in the phenotype. By analogy cellular phenotype should not be defined based upon the expression of individual RNAs but rather as dynamical changes in subsets of RNAs comprising selected RNA systems where the system‐associated RNAs are balanced with each other to produce the associated cellular function. Data to support these observations will be presented from live mouse and human cells, fixed tissue analysis and subcellular regional analysis. This idea provides a framework for understanding cellular heterogeneity in phenotypic response to various stimuli and provides a means for rethinking how to manipulate cellular responses so that desired outcomes are obtained including identification of molecules showing therapeutic efficacy.Support or Funding InformationNIH U01MH098953, NIH GM110005, NIH MH 110180, The Brain Research Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploring the biosynthesis of hypermodified bases one step at a time
Authors: Bandarian Vahe 
Published: June 2021

Transfer RNA (tRNA) is among the most highly modified biological molecules. To date, over 100 modifications have been documented in tRNA. Many are simple modifications, but a subset are hypermodifications that are introduced by complex, and in some cases, unknown enzymatic transformations. Our lab has been involved in deciphering the biosynthesis of queuosine and wybutosine, which are two broadly conserved hypermodified bases. This presentation will focus on recent biochemical and structural studies of the enzymes that catalyze key transformations in the biogenesis of these bases.Support or Funding InformationNIH GM72623This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
New technologies to interrogate G protein‐coupled receptor signaling
Authors: Kruse Andrew 
Published: June 2021

G protein‐coupled receptors (GPCRs) are the largest family of cell‐surface receptors in humans, and they play critical roles in processes ranging from neurotransmission to the maintenance of metabolic homeostasis. In order to better understand the molecular basis for GPCR responses to endogenous activators and synthetic drugs, we have developed a new method to track GPCR signaling in live cells using time‐resolved proximity labeling (GPCR‐APEX). This method reveals the collection of proteins a GPCR encounters following activation, and enables quantitative measurement of receptor fate following agonist stimulation. In addition to tracking GPCR signaling, we have developed new methods for the discovery of synthetic antibody fragments as modulators of receptor conformational change and signal transduction. Together with GPCR‐APEX, synthetic antibody fragment modulators allow investigation and control of GPCR signaling in new ways.Support or Funding InformationThis work was supported by grants from the Smith Family Foundation, The Vallee Foundation, and an NIH Early Independence Award (5DP5OD021345).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 Functional Role of Arginase 1 and Neutrophil Proteomics in Predicting Ischemic Stroke Outcome
Authors: Petrone Ashley Brooke , Steele Kelsey , Eisenman Rhae 
Published: June 2021

IntroductionPeripheral neutrophil (PMN) count is elevated within the first few hours of acute ischemic stroke (AIS), and neutrophilia is associated with greater AIS severity, larger infarct volume, and poor functional outcome. In animal models, PMN inhibition results in decreased infarct volume and improved functional outcome; however, this strategy has not been successfully translated in clinical trials. We hypothesize that PMNs are a heterogeneous population, consisting of both “beneficial” and “detrimental” subtypes, depending on their functional protein expression, and one PMN protein, arginase 1 (ARG1), may be a key contributor to detrimental PMN function post‐AIS. We have previously described a distinct temporal pattern of PMN expression following AIS between favorable and poor outcome AIS patients. Thus, the purpose of this study was to expand upon this examine the function relationship between ARG1, as well as other PMN proteins, and outcome following AIS.HypothesisWe hypothesized that PMNs isolated from AIS patients with poor outcome will have higher ARG1 expression compared to neutrophils in the favorable outcome group. Further, we hypothesize that PMNs isolated from AIS patients with poor outcome will have a distinct functional proteome, giving rise to altered or more aggressive function that leads to a poor outcome following AIS.MethodsWe quantified PMN ARG1 expression in a subset of 24 AIS patients – n=12 favorable and n=12 poor outcome. PMN ARG1 protein expression was determined by flow cytometry by co‐staining for the surface marker for PMNs, CD66b, and intracellular ARG1. Further, in a subset of these patients – n=3 favorable and n=3 poor outcome, we performed a proteomic analysis on isolated PMNs.ResultsARG1 protein expression was significantly higher in PMNs isolated from AIS patients with a poor outcome (mean ARG1 = 665 + 117) compared to AIS patients with a favorable outcome (mean ARG1 = 453+108) (p=0.008). Further, in the samples sent for proteomics, we observed several differences in the PMN proteome between the AIS outcome groups. Specifically, the PMN proteins ‐ ARG1, defensins 1 and 4, and protein s100a9 – were increased in the poor outcome compared to the favorable outcome group.ConclusionIn conclusion, targeting specific PMN proteins, such as ARG1, rather inhibiting PMNs as a whole, may represent a more targeted and successful approach to modulating PMNs post‐AIS to improve outcome.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pancreatic Neuroendocrine Tumors Require Homeostatic Signaling from the Unfolded Protein Response.
Authors: Oakes Scott A. , Moore Paul C. , Qi Jenny Y. , Warren Rachel , Thamsen Maike , Ghosh Rajarshi , Gliedt Micah J. , Hiniker Anne , Kim Grace E. , Maly Dustin J. , Backes Bradley J. , Papa Feroz R. 
Published: June 2021

The unfolded protein response (UPR) is an intracellular signaling pathway largely controlled by two ER transmembrane kinases—IRE1alpha and PERK—that communicate the protein folding status of the endoplasmic reticulum (ER) to the nucleus to maintain ER homeostasis. Hypoxia, nutrient deprivation, proteasome dysfunction, sustained demands on the secretory pathway or somatic mutations in its client proteins‐‐conditions often encountered by cancer cells—can lead to the accumulation of misfolded proteins in the ER and cause “ER stress.” Under remediable levels of ER stress, the UPR activates transcriptional and translational changes that promote adaptation (Homeostatic UPR). But when confronted with irremediable levels of ER stress, these adaptive measures fail, and the UPR instead switches strategies to trigger cell death (Terminal UPR).For the reasons mentioned above, ER stress is documented in many solid cancers, but whether ongoing UPR signaling is beneficial or detrimental to tumor growth remains hotly debated. Given their high secretory activity, we predicted that pancreatic neuroendocrine tumors (PanNETs) would be one neoplasm that is particularly sensitive to protein folding stress. Not only do PanNETs universally hypersecrete one more peptide hormone(s), but the endocrine cells of the pancreas from which these tumors derive are the cells in the body most impacted by genetic loss of the UPR in mice and humans. For the over 1,500 Americans diagnosed with a PanNET each year, surgery is the only potentially curative treatment. Unfortunately, the five year survival is extremely low for the ~25% of patients who develop metastatic disease. Hence, new targets are desperately needed for patients with advanced PanNETs.We find that the UPR is strongly unregulated in human PanNETs, and that these neoplasms are heavily reliant on elevated levels of Homeostatic UPR signaling to avoid the toxic effects of protein folding stress in vivo. Moreover, we discovered that targeted interventions to either reduce Homeostatic UPR outputs or alternatively trigger the Terminal UPR have potent antitumor effects in at least 2 distinct preclinical PanNET models. Inhibiting IRE1alpha or PERK leads to hyperactivation of the opposing arm and pushes it into a Terminal UPR, resulting in the observed anti‐PanNET effects in vivo. Together, these findings suggest that the UPR is a promising target in PanNETs and related neoplasms.Support or Funding InformationThis work was supported by the National Institutes of Health: R01DK095306 (S.A.O., F.R.P), R01CA219815 (S.A.O.), R01EY027810 (S.A.O., F.R.P.), and The Neuroendocrine Tumor Research Foundation (S.A.O.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of TGF‐beta induced Long Noncoding RNAs in Lung Cancer
Authors: Lai Tsung‐Ching , Hsiao Michael 
Published: June 2021

Lung cancer is the most lethal cancer worldwide due to high ratio of recurrence and high probability to undergo metastasis. As Epithelial‐mesenchymal transition (EMT) plays a major role during metastasis, how EMT is regulated by periphery in the lung cancer is particularly important. Although initiation of EMT can be triggered by TGF‐beta found to be overexpressed in lung cancer, how EMT can be affected by TGF‐beta signaling in different disease stages and its contribution to the heterogenecity of tumor cells are still unclear. This hampers our ability to effectively treat and block metastasis in lung cancer. Recently, long noncoding RNAs have emerged as a new player for tumor metastasis. We therefore hypothesize that lncRNAs mediated TGF‐beta signaling might be involved during EMT in lung cancer. Using RNA‐seq, we identified changes in lncRNAs profiles during different time points after TGF‐beta treatment in A549 cells. Early responded lncRNAs are major regulators in signaling and will be selected as primary candidates for further study. Expression patterns will be re‐validated by other lung cancer cell line with TGF‐beta treatment. The lncRNA candidates that have altered expression levels in response to TGF‐beta will be chosen to determine their effects on migration and gene regulation using wound healing and expression microarray. In situ hybridization will eventually be carried out to correlate the expression level of lncRNA in lung cancer clinical samples and re‐probe with EMT markers. The regulation of lncRNA in EMT would be validated with lncRNA knockout with TGF‐beta stimulation in A549 cells. The abilities of migration and invasion in knockout cells would be determined in vivo. In the end, we found the candidate lncRNA play a critical role in EMT with TGF‐beta stimulation in lung cancer.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular Mechanisms of Beta Cell Adaptation to Hyperlipidemia
Authors: Poffenbarger Mason C. , Lathen Daniel R. , Ricks H. Austin , Barlow Andrew M. , Roth Zoey F. , Bikman Benjamin T. , Tessem Jeffery Sivert 
Published: June 2021

Type 2 diabetes is characterized by hyperglycemia, hyperlipidemia and beta cell failure. Studies have shown that beta cell are extremely sensitive to hyperlipidemia, showing that short term cultures with elevated concentration of palmitate results in high levels of cell death. Though these studies demonstrate the toxic nature of palmitate on the beta cell, they do not accurately model the environment in which beta cells are exposed to elevated free fatty acids. We have, therefore, cultured the 832/13 beta cell line in progressively increasing levels of palmitate to determine the physiological and molecular changes that occur as beta cells adapt to this toxic environment. While 832/13 beta cells cultured immediately in 0.5mM palmitate result in sever cell death and failure to grow, 832/13 beta cells that are cultured in progressively increasing levels of palmitate (at 0.15mM, 0.3mM and 0.5mM palmitate) shows improved, although still impeded, proliferation rates. Culture in the progressively increasing concentrations causes downregulation of the beta cell transcription factors Nkx6.1 and Sox9, the glucose transporter Glut2, and the orphan nuclear receptors Nr4a1 and Nr4a3. Furthermore, as 832/13 beta cells are cultured for prolonged periods in elevated palmitate we observe downregulation of Nr4a target genes involved in mitochondrial respiration such as Idh3a, Idh3b, Idh3g, Dld and Dst. These changes correspond with modulation of mitochondrial respiration, and progressive deterioration of glucose stimulated insulin secretion, as well as decreased insulin content. These data begin to define the molecular changes that occur to the pancreatic beta cell as it is exposed to progressively increasing concentrations of palmitate, as is observed in type 2 diabetes disease progression.Support or Funding InformationThis study was funded by a BYU ORCA Grants to DRL, MCP, HAR and AMB. This study was also funded by a BYU MEG Grant to JST.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Epicatechin and its Gut Metabolites on Beta Cell Function, Survival and Proliferation
Authors: Tessem Jeffery Sivert , Rowley Thomas J. , Ray Jason D. , Herring Jacob A. , Kener Kyle B. , Bitner Benjamin F. , Ross Mimi , Lopez Moroni , Bikman Benjamin T. , Neilson Andrew P. 
Published: June 2021

An initiating factor of Type 2 diabetes (T2D) is beta cell dysfunction and the eventual loss of functional beta cell mass. Mechanisms that improve beta cell function could be used as a treatment for the millions of individuals with T2D. We have recently shown that the monomeric cocoa epicatechin is sufficient to enhance insulin secretion from the 832/13 INS‐1 derived beta cell line and primary rat islets through upregulating the Nrf2 pathway, enhancing mitochondrial protein expression, improving mitochondrial function and ultimately enhancing beta cell ATP production. Here we present data pertaining to the larger epicatechin forms (oligomeric and polymeric epicatechin) as well as gut microbe metabolites of epicatechin (Homovanillic Acid, Hippuric Acid, 5‐phenylvaleric Acid) on beta cell proliferation, survival and insulin secretion. While the monomeric form enhances insulin secretion, the oligomeric and polymeric forms inhibit insulin secretion. Interestingly, the gut metabolites of epicatechin all significantly enhance insulin secretion. These results suggest that cocoa derived monomeric epicatechin and its gut‐derived metabolites could be leveraged to enhance insulin secretion as a treatment modality for patients with T2D.Support or Funding InformationThis research has been funded by a grant to JST from the Diabetes Action Research and Education Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.