bluuebunny/crossref_metadata_2025_split huggingface dataset:
The MAL/SRF Pathway Regulates Desmosome Protein Expression and Localization in Cancer Cells
Authors: Dubash Adi D , Eldredge Luke 
Published: June 2021

The desmosome (DSM) is a cell‐cell adhesion complex required for the mechanical stability of tissues and regulation of other biological processes, such as proliferation and migration. Proteins from three families make up the complex: Transmembrane cadherins connect adjacent cells, plaque proteins stabilize attachment on the intracellular face, and the plakin protein desmoplakin (DSP) anchors the DSM complex to the intermediate filament cytoskeleton. Many studies have reported diverse mechanisms by which cancer progression alters DSM gene expression, but the signaling pathways that govern the expression and localization of DSM constituent proteins remain elusive. Recent work has identified the transcription factor serum response factor (SRF) as a regulator of mRNA levels and localization of DSM proteins such as the cadherin Desmoglein‐1. Here, we treated several cancer cell lines with siRNA and a pharmacological inhibitor (CCG‐1423) of SRF and its cofactor, MAL, to broadly investigate the role of these transcription factors in regulating DSM gene expression and protein localization. Our results demonstrate that abrogation of MAL/SRF signaling through both siRNA and pharmacological inhibition results in a decrease in mRNA levels of DSP. In addition, localization of DSP to borders is also reduced upon inhibition of MAL/SRF signaling. We observed similar changes for the plaque proteins Plakophilin‐2 and Plakoglobin, but not for desmosomal cadherins. As DSP is required to maintain the strength of DSM attachments, we also investigated the role of MAL/SRF signaling on the adhesion strength between cells. Our data highlight a novel link between MAL/SRF signaling and DSP expression, and contribute to a growing understanding of the variety of signaling pathways involved in mediating DSM gene expression.Support or Funding InformationThis project was funded by an NIH South Carolina IDeA Networks of Biomedical Research Excellence (SC INBRE) grant #5P20GM103499‐17 which provided a Developmental Research Project (DRP) sub‐award #22050‐ZA14 to AD.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Phylogenetics of Rhipicephalus sanguineus and its Role as a Vector of Rocky Mountain Spotted Fever
Authors: Schaefer Charles , Allen Jonathan , Yao Tony , Owen Haley , Lisowski Sasha , VandenBrooks John 
Published: June 2021

Rocky Mountain Spotted Fever (RMSF) is a tick‐borne disease caused by the intracellular bacteria Rickettsia rickettsii. The traditionally accepted vectors for this disease include ticks from the genus Dermacentor, however this genus is not endemic to the site of a recent outbreak in Arizona. Rhipicephalus sanguineus, also known as the Brown Dog Tick, is an ectoparasite that can be found throughout the United States. It has been hypothesized that this species exists as several distinct groups that form what can be more accurately described as a species complex. The Brown Dog Tick has been linked to this recent outbreak of RMSF in Arizona, despite no previous evidence of this species acting as a vector elsewhere in the US. Given the widespread habitat of the Brown Dog Tick, its potential as a vector for RMSF is a pressing public health risk. In this study, we sought to determine if differences associated with the genetic diversity of the Brown Dog Tick are responsible for its role as a vector. Ticks identified as R. sanguineous (n=241) were sorted into lineages based on phylogenetic analysis of a concatenated sequence of the 16S, 12S and COX1 genes. We used “Tropical” and “Temperate” marker sequences from identified samples on genbank (Table 1). Our results demonstrate the presence of multiple clades geographically distributed Arizona beyond just the “Tropical” and “Temperate” delineation. These tick samples were also tested for infection with R. rickettsii using PCR for the R. rickettsii OMPA gene. However, there was no significant difference between the phylogenetic tick clades (Chi‐squared, p=0.21). This suggests the genetic lineage of the vector tick is not the primary factor responsible for the outbreak of RMSF in Arizona and the surrounding regions.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Microbial Co‐occurrence Patterns and Keystone Species in the Gut Microbial Community of Mice in Response to Stress and Chondroitin Sulfate Disaccharide
Authors: Li Robert W. , Liu Fang , Li Xhaojie , Xue Changhu , Tang Qingjuan 
Published: June 2021

Detecting microbial interactions is essential to the understanding of the structure and function of the gut microbiome. In this study, microbial co‐occurrence patterns were inferred using a random matrix theory based approach in the gut microbiome of mice in response to chondroitin sulfate disaccharide (CSD) under healthy and stressed conditions. While the overall size and structure of the global networks were similar, the network composition was distinctly different among various experimental conditions. The exercise stress disrupted the network composition and microbial co‐occurrence patterns. 34 Operational Taxonomic Units (OTU) were identified as module hubs and connectors, likely acting as generalists in the microbial community. Mucispirillum schaedleri acted as a connector in the stressed network in response to CSD supplement and may play a key role for bridging intimate interactions between the host and its microbiome. Several modules strongly correlated with physiological parameters were detected. For example, Modules M02 (under stress) and S05 (stress + CSD) were strongly correlated with blood urea nitrogen levels (r = 0.90 and − 0.75, respectively). A positive correlation between node connectivity of the OTUs assigned to Proteobacteria with superoxide dismutase activities under stress (r = 0.57, P <0.05) provided further evidence that Proteobacteria can be developed as a potential pathological marker. Moreover, the results suggest that altered network topologies may have important functional implications. Our findings provided novel insights into microbial interactions in gut microbial communities of the mice in response to stress and CSD supplement and may facilitate future endeavor in microbial community engineering.Support or Funding InformationNational Natural Science Foundation of China (No. U1606403 to CX)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Preliminary Results of Assessing the Impact of a High School Science Outreach Program on the Confidence, Attitudes, and Skills of the Participants
Authors: Calderon Irene , Mina Rina 
Published: June 2021

Science outreach has been shown to enhance science education and cultivate students' interest in science. The Summit Country Day School, an independent Cincinnati‐based co‐educational K‐12 school, established a Milwaukee School of Engineering (MSOE) Center for Biomolecular Modeling Modeling a Protein Story (MAPS) team this school year, with the overall goal of increasing the scientific and technical skills of its participants. Through this 22 week program, the participants studied the structure‐function relationship of an assigned protein (Aquaporin) by researching relevant scientific articles and using software (Jmol) for molecular modeling to 3D print the protein structure. Seven underclassmen participated; two science faculty members facilitated and mentored. A Likert survey was developed to assess and compare the confidence, attitudes, and skills of the participants. Specifically, the tool evaluated these learning goals: 1) selecting relevant scientific articles, 2) assessing important points in the selected articles, 3) presenting the summary of the articles and their ideas to the group, 4) writing a scientific abstract, and 5) managing computer software for molecular modeling for 3D printing. The tool was administered at the beginning and halfway through the program, with planned administration at its conclusion. Wilcoxon signed‐rank test was performed to assess statistically significant differences between the survey responses at the two time periods. The survey findings demonstrated no statistically significant differences between the responses at the beginning of the program and at the halfway through the program. However, there were overall trends for improvement in the confidence, attitudes, and skills of the participants, especially regarding understanding scientific articles (Fig. 1). In conclusion, participating in a MAPS team generally increases the confidence, attitudes, and skills of the participants in regard to scientific activities. The lack of statistical significance may be because of the short duration of the program and small sample size.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TmAFP Interaction with an Ice Lattice
Authors: MacGillivray Brendan , Greig James , Bai Kathleen , Fu Celine , Fujikawa Ayaka , Hopkins Graeme , Illianu Richard , Kim Anna , Lalapet Tarun , Miao Crystal , Ramirez Victoria , Vo Minh Thu 
Published: June 2021

The mealworm beetle, Tenebrio molitor, has an antifreeze protein (TmAFP), which keeps it from freezing solid in sub‐zero temperatures. TmAFP prevents the growth of ice crystals by using repeating coils of a highly conserved 12‐amino acid loop sequence to bind to the ice surface. This structure creates a regular array of threonine‐residues which form specific hydrogen bonds with the ice lattice. As a small protein (8.4 kDa) with this repeating structure, TmAFP is among the simplest AFP structures to be crystallized. This structural simplicity may make it the perfect candidate to use as a tool in prolonging organs that have been harvested for transplant. A current problem is that organs are only viable for transplant for a day at the most. The best way to keep organs viable for prolonged periods of time would be to store them at freezing temperatures. However, if the organ freezes, ice crystals will kill cells and destroy the tissue. By introducing antifreeze proteins into organs before harvesting, the hope is to extend the lifespan of the organ for transplantation. The Ashbury College MSOE Center for BioMolecular Modeling SMART Team used 3D modeling and printing technology to examine structure‐function relationships between TmAFP and ice formation.Support or Funding InformationMSOE ‐ Center for Biomolecular Modelling ‐ co‐sponsor Diane MunzenmaierThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Flow Cytometry Analysis of Anastasis in Cultured Mammalian Lymphocytes
Authors: Moran Colleen Joy , Lauzon Robert 
Published: June 2021

Apoptosis is a physiological form of programmed cell death characterized by phosphatidylserine externalization, caspase activation, and genomic DNA fragmentation. Recent studies have shown that apoptosis may be reversed even in its late stages, calling the previous notion of a ‘point of no return’ into question. This recovery process has been termed anastasis (‘rising to life’), but its underlying mechanisms are not well understood. Our lab has developed a model of studying anastasis using cultured mouse HT‐2 lymphocytes dependent on the cytokine interleukin 2 (IL2). Our previous findings have shown that HT‐2 cells accumulate in the G1 stage of the cell cycle following 24 hours of IL2 deprivation and can be rescued by adding IL2 back into the culture medium. In this study, we have used quantitative flow cytometry to investigate which apoptotic events are congruent with anastasis. HT2 cells were deprived of IL2 for 24 hours and then rescued for periods of 0, 24, 48, and 72 hours. CD25 surface expression (high‐affinity IL2 receptor) and phosphatidylserine externalization (annexin V staining) were subsequently monitored prior to and following recovery, as markers of T cell activation and early stage apoptosis, respectively. Control HT2 lymphocytes continuously maintained in IL2 comprised a single viable population of CD25HIGHAnnexin−cells. Cells deprived of IL2 for 24 hours were observed to fall into two populations: CD25LOWAnnexin+early apoptotic and CD25LOWAnnexinnon‐apoptotic cells. In both populations, CD25 expression was reduced approximately 10‐fold when compared to IL2‐maintained controls. During anastasis, three distinct viable cell populations emerged: 1) CD25LOWAnnexin+cells that did not progress into the later stages of apoptosis, 2) CD25LOWAnnexin−non‐apoptotic cells, and 3) CD25HIGHAnnexin−non‐apoptotic cells. With advancing recovery time, viable CD25LOWpopulations decreased as CD25HIGHcells increased. By 72 hours post‐recovery, the majority of cells were actively cycling, annexin negative cells. These findings suggest that IL2‐dependent mouse HT‐2 lymphocytes may be a promising model to study the mechanisms of anastasis.Support or Funding InformationUnion College Faculty and Student Research GrantsThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Fibroblast Growth Factor 1 (FGF‐1) on CCN2 Gene Expression in Chondrocytic Cells
Authors: Elseoudi Abdellatif , ElKader Tarek Abd , Nishida Takashi , Eguchi Takanori , Aoyama Eriko , Hattori Takako , Takigawa Masaharu , Kubota Satoshi 
Published: June 2021

Cartilage and bone are developed by chondrocytes and osteoblasts differentiated from mesenchymal stem cells under the multiple interactions of extracellular signaling molecules. Among such molecules, CCN family protein 2 (CCN2) plays a critical role in the development and regeneration of the cartilage and bone to promote proliferation, differentiation and chondrocyte maturation as well as matrix remodeling during chondrogenesis. CCN2 interacts with several growth factors involved in endochondral ossification, which include fibroblast growth factor 1 (FGF‐1) showing a specific binding with this CCN2. However, the role of FGF‐1 in chondrocyte metabolism has not been investigated well. Therefore, in this study, we evaluated the effect of FGF‐1 on chondrocytes and its role on CCN2 regulation. The effect of FGF‐1 on CCN2 in human chondrocytic HCS‐2/8 cells and the cellular phenotype was estimated by the gene expression of chondrocytic markers. Addition of FGF‐1 to chondrocytic cell culture drastically repressed the mRNA levels of CCN2, with significant repression of other chondrocytic markers in those cells. The ELISA experiments revealed consistent results, in which the protein level of CCN2 was drastically down‐regulated by the addition of FGF‐1. Next, reporter gene assay was used to examine whether the CCN2 regulation by FGF‐1 is mediated by genetic elements involved in the proximal promoter, or not. However, the negative effect of FGF‐1 on CCN2 promoter activity appears to have been too weak to account for the observed decrease at mRNA and protein levels. Another reporter gene assay was performed with a plasmid containing multiple post‐transcriptional regulatory elements in the 3′‐untranslated region of the CCN2 gene. Surprisingly, FGF‐1 has no effect on these elements. Degradation fate of CCN2 mRNA after the addition of actinomycin D was tested, in the presence or absence of FGF‐1, showing unexpected results. Suspecting a regulation at a chromatin level, the effect of a histone deacetylase inhibitor, valproic acid (VPA), on the FGF‐1 function was evaluated, but VPA did not prevent the repressive effect of FGF‐1 on CCN2 gene expression. Further investigation is underway, suspecting a transcriptional regulation through a silencer located outside of the proximal promoter region.Support or Funding InformationJapan Society for the Promotion of Science (JSPS)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Genetic Variation of αENaC on Clinical Presentation in Cystic Fibrosis
Authors: Zavala Hanan , Dunitz Jordan , Sprissler Ryan , Snyder Eric 
Published: June 2021

ObjectiveIndividuals with the same disease causing variant of cystic fibrosis (CF) can have very different clinical outcomes, suggesting that yet‐to‐be identified modifier genes may play an important role in clinical presentation and disease progression. The epithelial sodium channel (ENaC) plays an important role in the pathophysiology of CF. Previous research has shown that substitution of alanine for threonine at position 663 of the SCNNA1 gene, which encodes the alpha subunit of ENaC, results in channel gain‐of‐function. In this study we sought to examine how genetic variation in the SCNNA1 gene influences clinical outcomes in CF.MethodsBuccal swabs were collected from thirty‐five CF patients that had at least one copy of the del.F508 mutation (19 males, 16 females). Samples were analyzed for allelic status at position 663 of SCNNA1 (AA, AT/TT). Medical charts were reviewed for resting vitals, pulmonary function, and indicators of disease progression.ResultsTwenty‐one subjects were homozygous for the alanine allele (AA, A663), 12 were heterozygous (AT) and 2 were homozygous for the threonine allele (TT). Due to low numbers in the TT group the AT and TT groups were combined for analysis (T663). Individuals in the T663 group had significantly higher resting heart rate (88±9 v 77±9, p=0.004) and tended to have lower body weight (133.7±56.5lbs v 143.6±31.9lbs), and higher systolic blood pressure (121±16.9mmHg v 115±16.5mmHg) compared to the A663 group. Forced vital capacity was significantly lower in the T663 group (3.64±1.12L v 4.29±0.94L, p=0.038). However FEV1/FVC was higher in the T663 group (75.0±12.1% v 71.8±11.0%, p=0.043), suggesting smaller lung volumes but not greater obstructive lung disease compared to the A663 group. There was no statistically significant difference in the frequency of pancreatic insufficiency or cystic fibrosis related type‐I diabetes between the two groups.ConclusionWe found that CF patients with the T663 allele tended to have smaller lungs as well as higher resting heart rate and blood pressure when compared to the A663 group. This finding suggests that the T663 variant results in cardiovascular compensation for altered pulmonary function.Support or Funding InformationNoneThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PM2.5 Activates Sirtuin‐SREBP1‐PIR Axis to Induce Pulmonary Fibroblast Inflammation: A Molecular Mechanism Model of Environmental Factor Induced Lung Fibrosis and COPD
Authors: Tien Chia‐Ping , Chang Yu‐Chan , Hung Shih‐Chieh , Hsiao Michael 
Published: June 2021

PM2.5 has been shown to have strong link with increased risk in human mortality and morbidity, including bronchitis, asthma, chronic obstructive pulmonary disease (COPD), and lung cancer. However, the underlying molecular mechanisms induced by PM2.5 remain unclear. To identify the molecular targets of PM2.5, we analyzed transcriptomics data by microarray chips in human pulmonary fibroblasts (HPF) after exposure to different doses of PM2.5. We have identified candidate genes that may involve in the induction of lung inflammation. Our results showed that the SREBF2 and SREBF1 are the top 2 transcription factors activated in response to PM2.5 exposure in HPF. Our analysis showed that IL‐6, IL24, LIF, PIR and HMOX1 were the most significantly downregulated transcriptional targets of SREBF1. Additionally, SREBP1, was found to be directly deacetylated and inhibited by SIRT1, as well as increase the expression of inflammasome markers, ex. NLPR3, ASC, and IL‐1β. Our results further showed that SREBF1, PIR, and NLRP3 inhibitors were found to reverse PM2.5 induced inflammation. In conclusion, our results identified that SIRT1‐SREBP1‐PIR/NLRP3 inflammasome axis may play a key role in PM2.5‐induced fibroblast‐mediated pulmonary disease and may serve as a new target for therapy to prevent fibrosis and COPD.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Detecting a Tumor Suppressor Protein (p16INK4a) in Zebrafish (Danio rerio)
Authors: Rodriguez Cody James , García Dana Michelle 
Published: June 2021

Zebrafish (Danio rerio) have been put forth as models for human disease since the 1960s. Zebrafish have also been identified as one of the most promising model organisms used to study aging. In this study, we are exploring a specific tumor suppressor protein p16INK4a which is encoded by the CDKN2A gene in humans. This tumor suppressor has been identified as a marker for cellular senescence in humans. Dysfunction of the protein has been associated with many cancers in humans, including leukemia, melanoma, squamous cell carcinoma, pancreatic, cervical, bladder, and even lung cancer. Studies locating this protein in adult zebrafish have not yet been accomplished in spite of the fact that the zebrafish version of the gene has been identified as cdkn2a/b. We hypothesize (1) that zebrafish synthesize the p16INK4a protein and (2) that the abundance of this protein correlates with aging. We used a rabbit monoclonal anti‐CDKN2A/p16INK4a antibody to label retinal sections taken from adult zebrafish eyes; Hoechst was used to label nuclei. Preliminary results suggest labeling of p16INK4a in the retina. Further experiments are underway to corroborate this initial finding. Localization of p16INK4a in zebrafish would support arguments for using zebrafish as a model organism in which to study phenomena associated with aging and cancer in humans.Support or Funding InformationCJR was funded by the US Department of Education HSI STEM program (84.031c), Award #P021C160036. CJR was also funded by Texas State University's undergraduate research fellowship fund (URF). Confocal images were obtained using Olympus FV‐1000 confocal microscope which is funded by NSF grant DBI‐0821252 to Dr. Joseph R. Koke and Dr. Dana M. García.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.