bluuebunny/crossref_metadata_2025_split huggingface dataset:
Investigation of Some Angiogenic Factors Related to Aging in Brown and White Adipose Tissues of Rats
Authors: Yurekli Muhittin 
Published: June 2021

Aging is a biological phenomenon in which life force decreases, there is an increase in dysfunction, and activities at the level of tissue and cell with insufficient response to changes in internal and external environments decrease. In biological aging, there are deficiencies in the cell level, the cells and tissues are prevented from feeding with enough oxygen and hypoxic conditions can occur. Due to the loss of function in living organisms as a result of aging, the failure of the organelles to perform their functions may lead to the emergence of oxygen deficiency conditions in the tissues. The most common feature of aging is the change of physiological functions over time. Especially when oxygen deficiency appears in the tissues, the number of red blood cells increases, hormones and angiogenic factors releasing increase. Hypoxia may also occur in tissues due to environmental factors, obesity and various circulatory insufficiency, and the organism may activate angiogenic factors in response to this. Adequate oxygen must be provided for the normal functioning of the tissues. In the case of oxygen deficiency (hypoxia), matrix metallo proteinase, vascular endothelial growth factor, tumor necrosis factor, interleukin may play role such as angiogenic factors. In this study, the levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase‐II, hypoxia‐inducible factor‐1α, adrenomedullin were investigated in 6 months and 24 months old rats. The vascularity function of the tissues is very important. Vein formation is required in both normal functional tissues and in tissues that are in neoplastic formation. Therefore, in this study, we investigated the angiogenic factors in rats' brown and white adipose tissue in cases by aging. In the study, 24 rats were divided into two groups; 12, 6‐month old young and 12, 24‐month‐old rats were evaluated as elderly group. In the study, angiogenic factors increased in the white adipose tissues of the elderly animals, whereas it was found that there was a greater increase in the brown adipose tissue that plays a role in the thermogenesis event in young rats. Decreasing physical activity in elderly animals and increased activity in white adipose tissue resulted in a significant increase in the weight of aged rats. Angiogenic factors play an important role in maintaining the oxygen demand of the organism in elderly animals, while it may cause an increase in the expanding of the white adipose tissue, which is effective on weight gain, which may lead to obesity.Support or Funding InformationThis research was funded by Inonu University, Department of Scientific Research Projects Project No: FYL‐2017‐748).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Breast cancer cell invasiveness is stimulated by loss of membrane interaction of actinbinding protein profilin1 via altered phosphoinositide metabolism
Authors: Roy Partha , Gau David , Bae Yongho , Ohayon Lee 
Published: June 2021

Profilin1 (Pfn1) belongs to a class of actin‐binding protein cytoskeletal function of which is negatively regulated by its interaction with membrane phosphoinositides (PPI). In stark contrast to most physiological scenarios where loss of Pfn1 significantly impairs normal cell migration and invasion through inhibition of actin polymerization, breast cancer cells exhibit hyper‐migratory phenotype in vitro and enhanced hematogenous dissemination from primary tumor in vivo upon depletion of Pfn1, the underlying mechanism of which is not easily explained by Pfn1's cytoskeletal function. Through knockdown and overexpression studies in multiple cell lines, we show that Pfn1 plays a key role in determining the cellular profile of various PPIs including PI(4,5)P2, PIP3 and PI(3,4)P2 Specifically, Pfn1 promotes PI(4,5)P2 and inhibits membrane accumulation of PI3K‐generated PPIs including PIP3 and PI(3,4)P2, a feature related to Pfn1's PPI interaction. Kinetics of PI3K signaling output suggest that loss of Pfn1 enhances sustenance of PI3K signaling by inhibiting an antagonistic pathway, a finding that correlates to Pfn1‐dependent regulation of PTEN. We establish Pfn1‐dependent alteration of multiple pro‐metastatic cytokines and membrane recruitment of selective PI(3,4)P2‐dependent pro‐migratory protein complexes, the latter in particular linking to hyper‐migratory and hyper‐invasive phenotypes of Pfn1‐deficient breast cancer cells. In summary, these findings establish novel regulation of cell migration by lipid signaling via membrane interaction of actin‐binding protein.Support or Funding InformationNIH 2R01CA 108607This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fatty Acids Bind to Them1, a Negative Regulator of Thermogenesis in Brown Adipose Tissue
Authors: Tillman Matthew C , Khadka Manoj , Okafor Chiamaka Denise , Juneja Puneet C , Ortlund Eric A 
Published: June 2021

Obesity and other metabolic diseases have become a global epidemic, greatly increasing in prevalence over the past 30 years. In mammals, brown adipose tissue counteracts obesity through thermogenesis: the burning of fat to produce heat. Thioesterase superfamily member 1 (Them1) is a multi‐domain enzyme in brown adipose tissue that slows down thermogenesis by converting the substrate acyl‐CoA into fatty acids. Additionally, Them1 contains a lipid‐binding StAR‐related lipid transfer (START) domain with an unknown function. We hypothesized that the lipid‐binding START domain controls thermogenesis by functioning as a lipid sensor for the enzymatic domains of Them1. We used mass spectrometry, X‐ray crystallography, and microscale thermophoresis to demonstrate that long‐chained fatty acids (14–20 carbon tail length), products of Them1's enzymatic reaction, bind to the START domain. Additionally, we determined a low‐resolution structure of full‐length Them1 using negative stain electron microscopy and found that Them1 forms a trimer, making contacts between the enzymatic domains. Using this structure, we ran molecular dynamic simulations to compare protein dynamics in the presence and absence of fatty acid. We determined that the largest differences in protein motions were localized to the START domain. In conclusion, we have used a robust, multidisciplinary approach to demonstrate that fatty acids, the product of Them1 enzymatic activity, bind the START domain, making Them1 the first protein of the START domain family identified to bind fatty acids. Additionally, we provide the first preliminary structure of full‐length Them1, which enhances our limited knowledge of multi‐domain thioesterases.Support or Funding InformationThis study was supported in part by the Emory Integrated Lipidomics Core (EILC), which is subsidized by the Emory University School of Medicine and is one of the Emory Integrated Core Facilities. Additional support was provided by the Georgia Clinical & Translational Science Alliance of the National Institutes of Health under Award Number UL1TR002378. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the 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.
MEK1/2 inhibitor reduces vascular calcification by regulating both canonical and non‐canonical Wnt signaling pathways
Authors: Zeng Peng , Yang Jie , Liu Lipei , Chen Yuanli , Han Jihong 
Published: June 2021

We previously reported that MEK1/2 inhibitors, such as PD98059 or U0126, synergized liver X receptor (LXR) ligand, such as T0901317, activated macrophage ATP‐binding cassette transporter A1 (ABCA1) expression and free cholesterol efflux. Therefore, the combined U0126 and T0901317 can regress the established lesions. Additionally, U0126 increased lesion stability with unknown mechanisms. Vascular calcification is a major risk factor for plaque rupture. In this study, we determined if MEK1/2 inhibitors can affect atherosclerotic calcification and the underlying mechanisms. In vivo, we determined that treatment of apoE−/− mice with U0126 reduced HFD‐induced atherosclerotic calcification. It also reduced calcification medium‐induced calcium deposition in aortic rings ex vivo or in human aortic smooth muscle cells (HASMCs). Associated with reduction of vascular calcification, expression of osteogenic genes, such as bone morphogenetic protein 2 (BMP2), runt‐related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), β‐catenin and osteopontin (OPN), were reduced by U0126, either in vivo or in vitro. Mechanistically, MEK1/2 inhibitors increased dickkopf 1 (DKK1) secretion and LDL receptor‐related protein 6 (LRP6) expression, thereby inhibiting canonical Wnt/β‐catenin signaling pathway while activating LRP6‐induced non‐canonical Wnt signaling pathway. In contrast, inactivation of either DKK1 or LRP6 attenuated the anti‐vascular calcification effects of MEK1/2 inhibitors. Based on the findings above, we conclude that MEK1/2 inhibitor can rescue vascular calcification through regulation of both canonical and non‐canonical Wnt signaling pathways. Our study also suggests inactivation of MER1/2 by chemicals can be a potential therapeutic strategy for treatment of atherosclerosis and atherosclerotic calcification.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Targeting Molecular Chaperone HSP90 To Treat Niemann‐Pick Type C1 Disease
Authors: Pipalia Nina H , Subramanian Kangaraj , Cross Abigail , Garg Kunal , Al‐Motawa Aisha , Maxfield Frederick R 
Published: June 2021

NPC1 is a fatal, neurodegenerative lysosomal storage disease caused by mutations in the NPC1 gene. More than 300 different disease‐associated mutations are reported in patients, resulting in abnormal accumulation of unesterified cholesterol, glycosphingolipids and other lipids in late endosomes/lysosomes (LE/Ly) of many cell types. There is no FDA‐approved therapy to treat NPC1 disease. A limited initial human trial of hydroxylpropyl beta cyclodextrin (HPBCD) is currently in progress. Our previous studies suggested that treatment with histone deacetylase inhibitors (HDACis) on skin fibroblasts results in increased expression of the NPC1 protein in mutant cells. Using an engineered human cell line, we have observed that HDACis can correct the NPC1 cholesterol storage defect in 60 of the 81 NPC1 mutants tested. However, when HDACis', were tested on non‐dividing cells like bone marrow derived macrophages, we did not observe any correction.Although, significant progress has been made in understanding the pathology of NPC1 disease, the mechanisms governing the proteostatic control of NPC1 protein is not well understood.. Disruption of molecular chaperone HSP90 (via hyperacetylation) is one of the many consequences of HDACi treatment. Thus, we evaluated the effect of direct inhibition of HSP90 family of proteins on NPC1 skin fibroblasts in terms of lowering lysosomal cholesterol accumulation.The data on the effect of several different HSP90 inhibitors on human patient‐derived skin fibroblast in lowering lysosomal cholesterol burden using microscopy based filipin assay will be presented. Our results suggests that cytosolic HSP90 and associated chaperones may be a target and that protein folding quality control plays a critical role in NPC1 diseases. We observed that out of the three HSP90 family proteins, inhibiting HSP90A and not HSP90B or TRAP resulted in clearance of cholesterol from lysosomes as well as aided in mobilizing unfolded mutant NPC1 protein from ER to lysosomes. We will also present the results elucidating the role of cytosolic HSP90 and its associated co‐chaperones in the regulation of NPC1 mutant protein stability.Support or Funding Information1. MDBR‐Orphan Disease UPenn Grant to Dr. Pipalia 2. APMRF Grant to Dr. Maxfield3. DART Summer Fellowship to Abigail CrossThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Vitamin E Reduces Hepatic VLDL Secretion by Disrupting its ER‐to‐Golgi Transport
Authors: Siddiqi Shadab A , Clay Ryan , Siddiqi Shaila 
Published: June 2021

Increased secretion of triglycerides rich‐very low‐density lipoproteins (VLDL) by the liver causes hyperlipidemia, which is associated with a variety of metabolic disorders. The rate‐limiting step in the secretion of VLDL particles from the liver is their transport from the endoplasmic reticulum (ER) to the Golgi and represents a potential therapeutic target in controlling VLDL secretion. Our laboratory has demonstrated that the targeted delivery of VLDL from the ER to the Golgi is facilitated by a distinct ER‐derived vesicle, VLDL transport vesicle (VTV). In this study, we investigated the role of vitamin E (alpha‐tocopherol) in the intracellular VLDL trafficking and secretion. To determine the role of vitamin E, we treated primary hepatocytes with different concentrations of vitamin E complexed with bovine serum albumin (BSA), carried out VTV‐biogenesis assays to examine ER‐to‐Golgi transport of VLDL and determined VLDL secretion. Our data demonstrate that vitamin E‐treatment of hepatocytes significantly reduces VTV‐biogenesis from the ER membranes and VLDL secretion. Interestingly, we found that vitamin E‐treatment significantly reduces the expression of proteins, Sar1b and SVIP, that control ER‐to‐Golgi transport of VLDL. We conclude that vitamin E controls VTV biogenesis by regulating the expression of Sar1b and SVIP proteins and thus plays an important role in VLDL secretion by the liver. This study was supported by NIDDK, NIH R01 DK081413 to SAS.Support or Funding InformationThis study was supported by NIDDK, NIH R01 DK081413 to SAS.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lysosome‐Associated Membrane Protein 1 Rescues Cholesterol Accumulation in NPC1 Mutant Cell by Enhancing Lysosome‐ER Contacts
Authors: Singhal Ashutosh , Song Byeongwoon 
Published: June 2021

Niemann‐Pick type C (NPC) disease is a neurodegenerative disorder caused by mutations in NPC1 or NPC2 gene, encoding a lysosomal membrane and luminal protein, respectively. NPC disease is linked to the disruption of intracellular cholesterol trafficking and is characterized by excessive accumulation of free cholesterol in lysosomes. The functions of NPC1 and NPC2 proteins are involved in the exit of lysosomal cholesterol by poorly understood mechanisms. Understanding the molecular mechanisms involved in lysosomal cholesterol export is critical for NPC therapy.We hypothesized that the lysosome‐associated membrane protein 1 (LAMP‐1) can mediate lysosomal cholesterol export in the absence of NPC1 function. By filipin staining, we demonstrated that 2‐hydroxypropyl‐β‐cyclodextrin (HPβCD) or HPγCD treatment rescues lysosomal cholesterol accumulation in NPC1 mutant cells derived from NPC patient. By proteomic analysis, immunoblotting, and confocal microscopy, we showed that HPβCD or HPγCD treatment in NPC1 mutant cells increases the expression of the LAMP‐1 protein and redistributes lysosomes toward the cell periphery. Overexpression and knockdown of LAMP‐1 in HeLa cells suggested that LAMP‐1 plays an important role in lysosomal cholesterol export. By confocal microscopy, we further demonstrated that LAMP‐1 is involved in establishing membrane contacts between lysosomes and the endoplasmic reticulum (ER). By analyzing the cholesterol content and proteome profile of isolated lysosomes, we provide new insights on the effect of HPβCD and HPγCD in modulating lysosomal functions and in reducing lysosomal cholesterol accumulation in NPC1 mutant cells.In conclusion, our studies suggest that LAMP‐1 can restore cellular cholesterol homeostasis by modulating lysosome positioning, potentially enhancing lysosome‐to‐ER cholesterol transport for cholesterol esterification and metabolism. Further studies on LAMP‐1 functions and lysosome dynamics are warranted to better understand NPC disease mechanisms and to design effective therapies for NPC.Support or Funding InformationThis study was supported by the Startup Fund, Pilot Grant, and Seed Grant to B.S. from Meharry Medical College.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluation of Dietary Protein Isolated from Different Sources on Hepatic and Adipose Lipid Metabolism
Authors: Warren Derek Cole , Skinner Roy Chris , Laub David , Barnes Kimbery , Matak Kristen , Tou Janet C 
Published: June 2021

Dietary protein is a significant contributor to the process of lipotoxicity through the modulation of insulin secretion and the regulation of adipocyte metabolic function. However, the contribution of different types of dietary protein is less understood. Previous studies have reported the effects of proteins isolated from multiple animal sources on liver lipid metabolism did not differ from casein; therefore, the objective of this study was to evaluate lipid metabolism in the liver and adipose in rats fed protein isolated from aquatic sources. Young (age 28 days) female Sprague‐Dawley rats were randomly assigned (n=8 rats/group) to standard purified diet with 10% w/w protein as casein, whey, Silver carp isolate or a protein‐free diet. There were no significant differences in liver weight or lipid content among rats fed these different protein sources. However, the protein‐free group had the highest hepatic lipid content (19.96% ± 4.13) among the diet groups confirmed by histological evaluation liver showing greater fat infiltration. Liver fatty analysis showed that rats on a protein‐free diet had the lowest (p<0.05) stearic acid content (6.77% ± 0.43), and the highest (p<0.05) oleic acid, linoleic acid, and α‐linolenic acid content at 20.81% ± 1.29, 23.21% ± 1.67, 1.56% ± 0.09, respectively. There were no differences in fatty acid composition in the liver of rats fed casein, whey, or carp isolate. However, rats fed whey isolate had greater (p<0.05) body weight and gonadal fat pad mass but no significant differences gonadal adipose fatty acid composition or serum LDL, HDL, and cholesterol compared to rats fed casein or carp. Collectively, these results indicated the amount rather than dietary protein source influences adipose tissue and hepatic lipid metabolism.Support or Funding InformationThis project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2015‐67017‐23084 from the USDA National Institute of Food and AgricultureThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Targeted Interleukin‐27 Gene Therapy Development for Disrupting Key Malignant Crosstalk between Metastatic Prostate Cancer and Bone
Authors: Salameh Janelle W. , Figueiredo Marxa L. 
Published: June 2021

Prostate cancer (PCa) bone metastases have been reported in up to 90% of patients with advanced disease. Prostate cancer cells ‘hijack’ bone homeostasis by disrupting the crosstalk between critical cells within the tumor/bone microenvironment (osteoblasts, osteoclasts, and immune cells), and utilizing this effector‐rich environment for cancer cell survival and growth. Crosstalk pathways primarily involve Interleukins (IL‐6 and IL‐11) and transforming growth factor β (TGFβ). Therefore, a key therapeutic objective in malignant skeletal disease management is to eliminate tumors while restoring bone homeostasis. Current treatments are limited to palliative radiotherapy, chemotherapy, or anti‐RANK treatments, all of which have considerable side effects such as osteonecrosis of the jaw or enhanced tumor invasion. An emerging approach to treating skeletal malignancies, osteoimmunology, investigates new multifunctional immune‐stimulatory agents that can simultaneously combat tumor growth, and promote bone regeneration. IL‐27 is an immunomodulatory cytokine with a great potential as a multifunctional cancer therapeutic. Its regulatory roles include antitumor immune responses (promoting Th1 differentiation), inhibition of tumor growth, and direct modification of tumor/bone crosstalk to support bone regeneration. Thus, we hypothesize that a tumor‐targeted IL‐27 gene therapy approach would facilitate the accumulation of IL‐27 in the bone/tumor microenvironment and enhance its therapeutic efficacy. To test this hypothesis, we created a tumor‐targeted IL‐27 gene by incorporating a novel peptide (LSLITRL); an IL‐6 receptor antagonist known to inhibit angiogenesis and tumor growth; or a scrambled control peptide at the C‐terminus. To detect the expression, secretion, and tumor recognition specificity, IL‐27 constructs were fused to a NanoLuciferase expressing vector. We have also assessed the potential for tumor‐targeted IL‐27 expression in disrupting malignant characteristics of PCa in vitro in terms of viability and invasiveness following transfection. Future studies will examine the expression and therapeutic efficacy of tumor‐targeted IL‐27 in vivo in a TRAMP‐C2 tibial implantation model.Support or Funding InformationNIH‐NCI R01CA196947‐S1This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Powerhouse of Depression: The Impact of MTCH‐1 Deficiency on Reward Signaling in C. Elegans
Authors: Penchina Kaitlyn , Zuklie Angus , Adams Clay , Dittmar Caleb , Park Madeline , Fong Megan , Langner Grace , Cole Austin , Madejski Sophia , Sweeney Robert , De Luke 
Published: June 2021

Major Depressive Disorder (MDD) is the leading cause of disability worldwide. One of MDD's most troubling symptoms is the loss of reward firing, or anhedonia. Current publications indicate that MTCH‐2, a gene associated with mitochondrial transport, is related to a decrease in firing rates in the hippocampus of mice. Additionally, a GWAS reported a link between MTCH‐2 and neuroticism in humans. Consequently, we hypothesize that a MTCH‐2 related decrease in firing rates in the nucleus accumbens could potentially play a role in the development of depressive symptoms.The C. Elegans MTCH‐1 protein shows a 87% homology to the human MTCH‐2 protein. Here, we examine the relationship between the MTCH‐1 gene and C. Elegans reward firing. Because C. Elegans reward firing levels have been associated with changes in speed change fluidity in past studies, we measure C. Elegans motility as a proxy for reward firing. In this study, we compare motility and gene expression in MTCH‐1 deficient worms and wt worms and further examine MTCH‐1 knockdown to control motility. Together, these data points may indicate a role of MTCH‐2 in depression.Support or Funding InformationThe Nueva SchoolThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.