bluuebunny/crossref_metadata_2025_split huggingface dataset:
NLRP3 Inflammasome regulation of lipolysis in aged adipose tissue
Authors: Camell Christina , Dixit Vishwa Deep 
Published: June 2021

Visceral adiposity is increased in the elderly and this increase is associated with increased risk for metabolic diseases. During aging, there are also increases in inflammation, senescence and leukocyte alterations in white adipose tissue. Furthermore, catecholamine‐induced lipolysis, the first step in generation of energy substrates through hydrolysis of triglycerides (TGs), declines with age. The defect in mobilization of free fatty acids (FFA) in elderly is accompanied with increased visceral adiposity, lower exercise capacity, failure to maintain core body temperature during cold stress, and reduced ability to survive starvation. While catecholamine signaling in adipocytes is normal in elderly, how lipolysis is impaired in aging remains unknown. Here we show that unlike diet‐induced obesity, aging is associated with reduction in adipose tissue macrophages (ATMs). We found ATMs regulate age‐related reduction in adipocyte lipolysis by lowering the bioavailability of norepinephrine (NE). Unbiased whole transcriptome analyses of ATMs revealed that aging upregulates genes controlling catecholamine degradation in an NLRP3 inflammasome‐dependent manner. Deletion of NLRP3 inflammasome in aging restored catecholamine‐induced lipolysis through downregulation of monoamine oxidase‐a (MAOA) that is known to degrade NE. Furthermore, inhibition of MAOA reversed age‐related reduction in adipose tissue NE concentration and restored lipolysis with increased levels of key lipolytic enzymes, adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL). Overall our findings suggest that NLRP3 inflammasome‐dependent immune‐metabolic interactions within AT contribute to age‐related inflammation and metabolic dysfunction. Our study also reveals that targeting neuro‐innate signaling between sympathetic nervous system and macrophages may offer new approaches to mitigate chronic inflammation‐induced metabolic impairment and functional decline.Support or Funding InformationThis work was funded in part by a supplemental award from the National Institute of Aging (grant number: 3R01AG043608‐03S1 FAIN:R01AG043608), the Irene Diamond AFAR Postdoctoral Transition Award, and K99AG058800This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression and purification of N‐terminally acetylated Ssa1 chaperone from Saccharomyces cerevisiae
Authors: Griffith Alijah A. , Holmes William 
Published: June 2021

N‐terminal acetylation leads to diversity of a given organism's proteome through constituting a cell‐wide acetylome wherein polypeptides exhibit an altered function and dynamic, acting as a basis for cell signaling pathways and allowing a given cell to react specifically to environmental stressors or cellular perturbations. The reaction of N‐terminal acetylation is catalyzed by a collection of N‐terminal acetyltransferases (NATs), wherein NAT complexes transfer acetyl groups from acetyl‐coenzyme A to the a‐amino group at amino‐terminal residues, effectively neutralizing the positive charge of the amino residue. One potential target of N‐terminal acetylation is thought to be molecular chaperone Ssa1. Endogenous to the eukaryotic Saccharomyces cerevisiae, Ssa1 is HSP70 class molecular chaperone that works in tandem with its co‐chaperone Sis1 to maintain cellular proteostasis through restoring native conformations of misfolded proteins through a transient mechanism dictated by ATP hydrolysis. Overall, the consequences of N‐terminal acetylation on the Ssa1 function and ability to maintain cellular proteostasis remains to be fully characterized. To further probe the functional and mechanistic changes associated with the N‐terminal acetylation of Ssa1, we developed an efficient, user‐friendly, one‐step purification of Ssa1 utilizing a previously constructed Protein‐A transformant strain. Our results indicate that our method is able to natively isolate high yields of pure Ssa1 from small amounts of starting material in a short period of time while allowing for the native elution of Ssa1 without compromising enzymatic activity, allowing the eluted protein to be utilized in downstream analyses to better understand interactions between Ssa1 and its co‐chaperones as well as the effects of post‐translational modifications on chaperone function.Support or Funding InformationResearch supported by RI‐INBRE (#P20GM103430) from NIGMS, NIH.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Methionine sulfoxide reductase reverses oxidation of the nucleotide exchange factor Fes1 to regulate cytoplasmic Hsp70 chaperone cycle
Authors: Nicklow Erin E , Kim Jinnie , Sevier Carolyn S 
Published: June 2021

Methionine residues in proteins are targets of oxidation by reactive oxygen species (ROS). Selective oxidation of methionine to methionine sulfoxide (MetO) can allow for reversible, beneficial modification of proteins to limit cell damage during conditions of excess ROS. Methionine sulfoxide reductases (Msrs) are conserved enzymes that reduce MetO residues in proteins, serving to both limit the over‐oxidation of protein methionines, and to remove MetO adducts when stress subsides. Although loss of Msr function has been implicated in neurodegenerative disorders and aging, it is unclear how Msr activities influence these disease pathologies because the vast majority of their physiological substrates remain unknown.We have identified the cytoplasmic Hsp70 nucleotide exchange factor Fes1 as an Msr substrate in yeast. Under oxidative stress, Fes1 is oxidized at a cluster of N‐terminal methionine residues. Current data show that this Msr‐controlled modification reversibly regulates interaction between Fes1 and Hsp70, thereby modulating chaperone nucleotide exchange, ATP hydrolysis, and peptide binding and release. We suggest that these altered Hsp70 activities, as a consequence of Fes1 modification, may help cells cope with elevated ROS by limiting peptide aggregation and/or activating cellular stress response pathways, whereas sustained modification of Fes1 in the absence of Msr activity is detrimental to proteostasis under non‐stress conditions. Moreover, our studies with Fes1 uncovered that the two yeast Msr proteins (Mxr1 and Mxr2) are expressed in the cytoplasm, in contrast to prior studies of Mxr2 suggesting sole mitochondrial localization. We show that Mxr1 and Mxr2 co‐operate to regulate the redox state of Fes1 both in vivo and in vitro. Together, these studies provide a new perspective on how activities of the well‐studied Hsp70 family can be modulated by ROS, and further define how methionine modification is used as a means for cells to sense and respond to oxidative stress.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Auto‐proteolysis of Type 1 Metacaspases in Schizophyllum commune
Authors: Cummings Brianna Paige , Fox Kristin M. 
Published: June 2021

Apoptosis is a specialized form of programmed cell that occurs under conditions such as stress, aging, and development. A key marker of apoptosis is the presence of caspases that cleave peptide bonds after aspartate residues via a catalytic histidine/cystine dyad. A group of related enzymes called metacaspases are found in plants, protozoa and fungi. Like caspases, they contain an active site histidine/cystine dyad and have a similar protein fold, yet they show preference for lysine and arginine residues and many require calcium for cleavage. Furthermore, Type I metacaspases contain a proline‐rich N‐terminal prodomain that is typically cleaved to achieve the fully active form. In some Type I metacaspases, continued auto‐proteolysis is observed after the prodomain is removed.Within the fungus Schizophyllum commune genome five type I metacaspases (ScMC1‐5) have been characterized and expressed with and without the prodomain. Additionally, optimal pH and calcium concentrations have been determined for each of these metacaspases. To further understand the auto‐proteolysis of these metacaspases, attention was focused on ScMC1. A time course has been established over which ScMC1, in the presence of calcium, auto‐processes to form a 40kDa fragment. Fluorescent activity assays over the same time course have shown a decrease in overall activity on small peptide substrates of the incubated enzyme. Additionally, the H211Q/C267A active site double mutant form of ScMC1 showed decreased activity and no auto‐proteolysis upon incubation with calcium. Lastly, incubation of wild‐type ScMC2‐4 with the double mutant ScMC1 showed no proteolysis of ScMC1, indicating cleavage of ScMC1 by other Type I metacaspases does not occur. These results have furthered our understanding of metacaspase activation and function, and ultimately their role in fungal apoptosis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Integration of signaling pathways and physical forces during vascular growth
Authors: Iruela‐Arispe Luisa , Aragon Raquel , McDonald Austin 
Published: June 2021

Our understanding of vascular morphogenesis and angiogenic growth has progressed considerably during the last few decades. Today the field has clarified the framework of interacting signaling pathways, intracellular regulatory genes and participating physical forces involved in the process of endothelial cell sprouting, lumen formation, and vascular remodeling. Nonetheless, once formed, vascular tubes also expand in width and length within the context of shear stress and tensional forces. Currently little information is available on how blood vessels perform this task or even what is the average half‐life of endothelial cells in the vascular wall. Using a combination of EdU incorporation, genetic tracing, rigorous quantification, and SILAM proteomics, our laboratory has gathered substantial information that sheds light into the cellular mechanisms and molecular players involved. The questions that will be addressed in this talk include: How does a vessel like the aorta expand in length and width while exposed to multiple physical forces? Do endothelial cells in an adult vessel divide to renew the endothelial lining or do [some] endothelial cells last for the lifetime of an individual? If endothelial cells proliferate: Is there a particular site/niche or do all cells all have equivalent capacity? Do endothelial cells have a finite number of cell divisions? What is the half‐life of endothelial cells in adult, normal blood vessels? What are the regulatory mechanisms involved in promoting vascular expansion? Taken together, we view this information as fundamental to our understanding the mechanisms controlling endothelial regeneration and their deregulations in settings like chronic/acute inflammation, aging, chronic diseases and physical trauma.Support or Funding InformationNIH/NHLBI R35HL140014This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Discovering novel enzymes and metabolic pathways
Authors: Gerlt John 
Published: June 2021

The number of proteins in the UniProt database (>127M sequences in Release 2018_08) is increasing with a doubling time of 2.5 yrs; however, ≥50% of the proteins have uncertain, unknown, or incorrect functions. The lecture will describe publicly accessible (“democratized”) web tools for protein sequence similarity networks (SNNs; EFI‐EST; efi.igb.illinois.edu/efi‐est/), 2) genome neighborhood networks (GNNs; EFI‐GNT; efi.igb.illinois.edu/efi‐gnt/), and 3) chemically guided functional profiling (EFI‐CGFP). These tools are used to 1) analyze sequence‐function space in protein families (EFI‐EST), 2) identify functionally linked enzymes in metabolic pathways (EFI‐GNT), and 3) prioritize protein families for functional assignment based on metagenome abundance (EFI‐CGFP). Examples will be described of the use of SSNs, GNNs, and CGFP to discover novel metabolic pathways in diverse microbial species, including the human gut microbiome.Support or Funding InformationNIH/NIGMS P01GM118303This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EFFECT OF PHOTOBIOMODULATION ON MUSCLE REPAIR AND OXIDATIVE STRESS CAUSED BY BOTHROPS JARARACUSSU VENOM
Authors: Silva Luciana Miato Gonçalves , Freitas Sarah Cristina Ferreira , Zamuner Luis Fernando , Cogo José Carlos , d'Angelis Katia , Zamuner Stella Regina 
Published: June 2021

INTRODUCTIONEnvenoming caused by snakes of the genus Bothrops is often associated with severe and complex local pathological manifestations, with dermonecrosis and myonecrosis being the most severe damage. The specific treatment for Bothrops accident is the use of the antivenom, with the function to neutralize the circulating venom, but its action does not extend the local manifestations. Previous studies in our group have shown that photobiomodulation using low intensity laser (LBI) increases the viability of C2C12 muscle cells when incubated with B. jararacussu venom (BjV) and further promotes the proliferation of these cells.AIMto analyze the effect of LBI on muscle repair and the oxidative stress caused by B. jararacussu venom on C2C12 myoblast cells.METHODOLOGYMyoblasts cells were plated at 1×104 cells/well and incubated for 24 hours for cell adhesion. BjV (12.5ug/ml) was applied and the cells were immediately irradiated with LBI (660 and 780 nm, power of 100 mW, irradiated time 10 sec, energy density of 4 J/cm2) and incubated for 1 h. Then, a wound was created by scratching cells with a sterile pipette tip and the cell migration into the wound surface was analyzed 24 h after the venom incubation. Oxidative stress (NO, H2O2, SOD, Gpx and TBARS) were analyzed 1 h after BjV incubation. The NO concentration was evaluated by the GRIESS and the H2O2 by the phenol red method. The concentration of SOS and Gpx was made by spectrometry. The analysis of TBARS by the Elisa method.RESULTSResults showed that the LBI was able to almost completely close the risk of the wound, similar to the control group. C2C12 cells were able to produce both NO and H2O2 one hour after the addition of the venom being statistically different from the control. LBI irradiation significantly reduced NO release, but did not alter the release of H2O2. No significant difference was observed of anti‐oxidant enzymes in C2C12 cells incubated to BjV. LBI Irradiation induced a significant increase of SOD in comparison to the venom group, but not modify Gpx release. BjV did not cause lipid peroxidation and LBI treatment did not modify this effect.CONCLUSIONPhotobiomodulation caused muscle cell protection and this protection seems to be related to decrease of reactive species of oxygen and nitrogen and increase of anti‐oxidants.Support or Funding InformationComissão de Aperfeiçoamento de Pessoal do Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Sirt3 in Lipid Generation
Authors: Le Truc P , Ma Oanh , Talbott George , Ho Linh 
Published: June 2021

Sirt3 is a major mitochondrial deacetylase which has important roles in regulation of metabolic homeostasis and metabolic diseases. ST2 cells are mesenchymal stem cells taken from bone marrow. The purpose of this research is to identify if Sirt3 plays a role in marrow adipogenesis to regulate insulin sensitivity through adipokines, especially adiponectin using ST2 cell models.Particularly, we used the gain‐function of Sirt3 in ST2 cells to assess the role of Sirt3 in adipogenesis. We were able to successfully overexpress Sirt3 and differentiate ST2 cells into adipocytes. We found that the induction of Sirt3 leads to increased adipogenesis compared to controls. The increase in adipogenesis was in line with elevated triglyceride levels in Sirt3 overexpression compared to controls (P < 0.01 in day 5 and P < 0.05 in day 7 after differentiation). This data is consistent with the increase in percentage area of adipocytes, which was highly significant in Sirt3 overexpression than that in the controls using Oil Red O staining (P< 0.01). The induction of adipogenesis by Sirt3 is associated with increased gene expression of adipocyte markers as well as adiponectin/adipokines.In addition, we inhibit Sirt3 function in ST2 cells using an inhibitor of Sirt3 called 3‐TYP. We treated cells with 50μM and 100M of 3‐TYP, versus control. Our results show that ST2 cells treated with Sirt3 inhibitor decreased adipogenesis by reduced area of adipocytes compared to controls especially at 100μM of 3‐TYP (P<0.05). Triglyceride measurements of 3‐TYP treated cells versus controls showed consistent results with reduced adipogenesis, especially at 100 μM on day 5 (p‐value <0.001). The expression levels of adipocyte markers and adiponectin/adipokines were statistically significantly reduced compared to the controls.These statistically significant consistent data suggest that Sirt3 gene plays an important role in modulating adipogenesis and adiponectin/adipokines expression. We hope our study contributes to the efforts of revealing Sirt3 functions in metabolic homeostasis and diseases. The ultimate goal is to use Sirt3 as a potential target in treating insulin resistance and other metabolic abnormalities.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Functional analysis of p53 response elements in the human Polycystic Kidney Disease Gene 1 (PKD‐1) promoter
Authors: van Bodegom Diederik , Roessingh Wijnand , El‐Dahr Samir 
Published: June 2021

We have previously shown that p53 represses human PKD1 expression in vivo and in vitro, and that this effect is dependent on histone deacetylase activity (JBC, 281, p.31234, 2006). The 5′‐regulatory region of PKD1 contains 4 putative p53 response elements (p53RE4‐1) at positions: −2.7kb, −1.2kb, −0.8kb and −0.2kb respectively. In this study, we delineate the differential roles of the p53RE's in transcriptional repression of the PKD1 gene. We performed site‐directed mutagenesis of distal p53RE4, proximal p53RE1. The wild‐type and mutant PKD1 promoter‐reporter constructs were co‐transfected with a p53 expression plasmid in IMCD3, HCT116(p53+/+) and HCT116(p53−/−) cells. p53RE4 deletion decreased PKD1 baseline promoter activity (2.4, 1.8 and 1.4‐fold) in IMCD3, HCT116(p53+/+) and HCT116(p53−/−) cells, respectively, and significantly enhanced p53‐mediated repression in all cell lines, indicating that p53RE4 is a positive cis‐response element. In contrast, deletion of the p53RE1 induced a 1.6‐fold increase in baseline PKD1 promoter activity; this was observed in p53+/+ but not p53−/ − cells. p53RE1 failed to mediate repression of a known p53‐target gene ‐the bradykinin B2 receptor‐ indicating that p53RE1 functions in a context‐specific manner. Chromatin Immunoprecipitation assays in IMCD3 cells revealed the DNA regions surrounding p53RE1/2 are occupied by HDAC 1 and 2. We conclude that the balance between p53RE2 and p53RE1 occupancy by p53 and HDACs determines the final response of PKD1 to p53 within a given cell.Supported by NIH grant DK‐062250
Regulation of heme degradation in association with multiple sclerosis
Authors: Kralik Lubomir , Krasulova Eva , Hrdinka Matous , Pospisilova Lenka , Tyblova Michaela , Havrdova Eva , Martasek Pavel 
Published: June 2021

Heme oxygenase‐1 (HO‐1) is a stress protein that degrades heme to biliverdin, iron, and carbon monoxide (CO). A (GT)n dinucleotide repeat polymorphism in the HO‐1 promoter has been shown to modulate HO‐1 gene expression. Short (<25) GT repeats are associated with HO‐1 up‐regulation. Multiple sclerosis (MS) is an autoimmune disorder resulting in CNS demyelination and neurodegeneration. HO‐1 or exposure to its end product CO counters autoimmune neuroinflammation and might have therapeutic potential for the treatment of MS. In this study, we investigated the association of the HO‐1 promoter (GT)n repeat polymorphism in 291 patients with MS. We used data from previous studies as a control group. We compared the frequencies of short (<25) repeat (class S) and long (≥25) repeat (class L) alleles. No significant difference between these two groups was found. Thus, the (GT)n dinucleotide repeat polymorphism in the HO‐1 promoter is not associated with MS. Comparison of the course of MS disease in patient groups homoallelic for either S/S or L/L may add yield differences in the expression of the diseases symptoms, suggesting that therapeutic intervention with HO‐1 inducers may be possible.Supported by Grants: IGA MZ 1A8713‐5, GAUK 109/2006, MSMT 0021620806.
Effect of fucoidan on mRNA and promoter activity of matrix metalloproteinases(MMPs) in skin fibroblasts
Authors: Moon Hee Jung , Shim Sun ‐ Nyu , Ku Mi‐Jeong , Lee Yong‐Hwan 
Published: June 2021

In the present study, to find molecular mechanism of UVB induced matrix metalloproteinase (MMPs) in skin fibroblasts, we investigated the effect of fucoidan on the MMP‐1 mRNA level and promoter activity. Fucoidan treatment of 0, 1, 10, 100 μg/mL concentration. The cells were incubated for 24 hr and after UVB irradiation, they were then further harvested. mRNA expression of MMPs was estimated by using RT‐PCR. The cells pretreated with fucoidan were inhibited in the expression of MMP‐1 at mRNA levels in the following order; 100 μg/mL> 10 μg/mL > 1 μg/mL compared to the control. In order to study the inhibitory effects of fucoidan on UVB‐induced MMP‐1 expression at transcription level, luciferase assay was performed using pGL3 MMP‐1 construct. To examine the roles of UVB in MMP‐1 promoter activity, a luciferase assay was performed at 0, 6hr, 12hr and 24hr after UVB (100 mJ/cm2) exposure. MMP‐1 promoter activity was maximally induced at 6hr after UVB irradiation and subsequently declined to basal levels in cells.In conclusion, fucoidan may down‐regulated MMP‐1 promoter and induce inhibition of MMP‐1 mRNA. Through this mechanism, fucoidan may have anti‐photoaging effect.*This work was supported by the grant of KICOS
Conditional knock‐down of the transcriptional coactivator PGC‐1α in mice using Cre‐LoxP induced RNA interference
Authors: Bagattin Alessia , Hugendubler Lynne , Hayes Schantel , Mueller Elisabetta 
Published: June 2021

PGC‐1α (peroxisome proliferator‐activated receptor gamma coactivator 1α) is a transcriptional coactivator involved in mitochondrial biogenesis and energy metabolism. The function of PGC‐1α has been studied in vivo through the generation of tissue‐specific transgenic and knock‐out models. We have used RNAi technology to generate transgenic mice with a conditional knock‐down of PGC‐1α in order to study the contribution of PGC‐1α in the response to various metabolic challenges. Mice carrying the U6‐ploxPneo‐PGC‐1α‐RNAi transgene were crossed with EIIa‐Cre transgenic mice to ubiquitously knock‐down PGC‐1α expression in vivo. U6‐PGC‐1α‐EIIa‐Cre mice were born at the expected Mendelian ratio. Upon cold exposure, PGC‐1α knock‐down transgenic mice exhibit reduced PGC‐1α mRNA induction in brown fat compared to controls and, as previously shown in knock‐out mice, they have decreased UCP1 (uncoupling protein 1) mRNA levels. Furthermore, microscopic analysis of H&E stained brown fat tissue sections revealed accumulation of large lipid droplets, as previously observed in PGC‐1α knock‐out animals. In conclusion, we have developed an animal model that mirrors the total PGC‐1α knock‐out mouse and that allows investigation of PGC‐1α function in vivo through a system that does not require the lengthy process of gene‐targeting nor ES cell manipulation.Supported by the Intramural Research Program of NIDDK, NIH.