bluuebunny/crossref_metadata_2025_split huggingface dataset:
DELETION OF MICRORNA‐22 ENHANCES THERMOGENIC GENE EXPRESSION IN WHITE ADIPOSE TISSUE OF OBESE MICE
Authors: Lima Vanessa Morais , Fonseca Renata Inzinna , Wang Da‐Zhi , Diniz Gabriela Placona 
Published: June 2021

BackgroundObesity is a major risk factor for several cardiovascular and metabolic disorders. In the last years, diverse studies have suggested that browning of white adipose tissue (WAT) may be used as a novel strategy for treatment of diseases associated to obesity. We recently reported that loss of microRNA‐22 attenuated the fat mass gain induced by high fat (HF) diet. In addition, loss of microRNA‐22 enhanced energy expenditure in mice fed a HF diet, suggesting that microRNA‐22 may play a critical role in the control of metabolic disorders observed in obesity. However, the possible mechanisms by which microRNA‐22 mediates the gain of WAT in response to HF feeding are unclear. In the present study, we investigated the contribution of microRNA‐22 in browning of WAT.MethodsMale microRNA‐22 knockout (miR22KO) and wild type (WT) mice were fed a chow diet or HF diet for 12 weeks. Body weight gain was monitored weekly. Epididymal fat was collected and expression of genes involved in browning of WAT was evaluated by qPCR.ResultsmiR22KO and WT mice fed HF diet displayed increased body weight gain when compared to their respective controls fed a chow diet. Loss of microRNA‐22 did not attenuate the body weight gain mediated by HF diet. Both miR22KO and WT mice fed HF diet exhibited increased epididymal fat mass compared to their respective controls. However, deletion of microRNA‐22 attenuated the gain of fat mass induced by HF diet. Analysis of gene expression demonstrated that miR22KO mice fed either a chow or HF diet displayed higher expression levels of UCP1 in WAT. In addition, miR22KO mice fed HF diet exhibited increased expression levels of Cidea and Prdm16 in WAT when compared to WT mice fed HF diet.ConclusionsThese findings suggest that loss of microRNA‐22 induces thermogenic gene expression in WAT of obese mice.Support or Funding InformationFunding: Sao Paulo Research Foundation (FAPESP #2015/21859‐5; #2017/01558‐6).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Melatonin or Dark Therapy Reduce Biliary Damage, Inflammation and Liver Fibrosis in a Murine Model of Early Stage Primary Biliary Cholangitis
Authors: Kennedy Lindsey , Wu Nan , Francis Heather , Venter Julie , Meng Fanyin , Kyritsi Konstantina , Zhou Tianhao , Gaudio Eugenio , Glaser Shannon , Alpini Gianfranco 
Published: June 2021

BackgroundPrimary biliary cholangitis (PBC) is characterized by increased biliary damage, inflammation and liver fibrosis. Early stage PBC is marked by biliary proliferation, whereas late stage PBC shows ductopenia. We have shown that dominant‐negative transforming growth factor b receptor II (dnTGFβRII) mice at 12 wk of age mimic early stage PBC. We have found that melatonin therapy or prolonged exposure to complete darkness reduces biliary hyperplasia and liver fibrosis in models of cholestasis. However, the impact of melatonin or dark therapy on PBC‐related injury is unknown. The aim of our study was to evaluate the effects of melatonin or dark therapy in a mouse model of early stage PBC.MethodsWe used background‐matched, male wild‐type (WT) and dnTGFβRII at 12 wk of age that were given drinking water containing melatonin (0.03%) or placed in complete darkness for 1 wk along with the related controls. Liver damage was evaluated by H&E. Intrahepatic bile duct mass (IBDM) was measured by CK‐19 staining. Biliary senescence was evaluated by staining for p16 and p21, and SA‐β‐galactosidase activity. Biliary and liver inflammation were determined by IL‐6 and F4/80 (Kupffer cell marker) staining. Liver fibrosis was determined by Sirius Red staining and immunofluorescence for collagen type‐1a. Hepatic stellate cell (HSC) activation was shown by SYP‐9 and α‐SMA staining. Human control and early stage PBC serum samples were obtained, and serum melatonin levels were measured by EIA.ResultsdnTGFβRII at 12 wk of age (early stage PBC mouse model) treated with melatonin or complete darkness had decreased (i) IBDM, (ii) biliary senescence, (iii) liver fibrosis, (iv) biliary and liver inflammation, and (v) HSC activation/liver fibrosis. Human early stage PBC samples had decreased serum melatonin levels.ConclusionInhibition of melatonin signaling perpetuates biliary damage associated with PBC. Restoration of melatonin‐dependent signaling via melatonin treatment or dark therapy may be therapeutic for patients with early stage PBC.Support or Funding InformationNIH NIDDK R01, VA MeritThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Calreticulin stabilizes vascular endothelial growth factor‐A mRNA via interaction with AU‐rich element at 3′‐UTR
Authors: Chen Chih‐Yu , Chien Yin‐Chieh , Lee Hsinyu 
Published: June 2021

Messenger RNA (mRNA) stability is a post‐transcriptional regulation that determines mRNA fate and influences multiple cellular functions. Previous studies have demonstrated that AU‐rich element (ARE) in 3′‐untranslated region (UTR) of mRNA is critical for mRNA stability via interaction with ARE‐binding proteins (ARE‐BPs). Chaperone protein calreticulin (CRT) has been reported to bind the 3′‐UTR, suggesting that it may act as a novel ARE‐BP. The expression level of CRT positively is correlated with the expression of angiogenic factor vascular endothelial growth factor‐A (VEGF‐A). However, the detail regulatory mechanism of CRT on VEGF‐A expression remains elusive. Therefore, we aimed to investigate whether CRT regulates mRNA stability of VEGF‐A through interaction with ARE. Our results showed that depletion of CRT significantly destabilized VEGF‐A mRNA and subsequently decreased the expression levels of both VEGF‐A RNA and protein in PC3 human prostate cancer cell line. Since several AREs were identified in the mRNA of VEGF‐A, we further explore the relationship between CRT and these AREs. Pull‐down analysis demonstrated that CRT directly interacts with the ARE region of VEGF‐A mRNA. In addition, we constructed luciferase vectors contained the ARE of VEGF‐A and co‐transfected with siRNA of CRT or CRT overexpression vector. Our results demonstrated that CRT interacts with ARE in 3′‐UTR of VEGF‐A mRNA. In summary, this study suggests that CRT stabilizes VEGF‐A mRNA through interacting with ARE at the 3′‐UTR of VEGF‐A mRNA.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Human Neural Progenitor Cells and Neurons Differentiated from Induced Pluripotent Stem Cells
Authors: Apáti Ágota , Berecz Tünde , Szabó Eszter , Orbán Tamás István , Homolya László , Sarkadi Balázs , Réthelyi János 
Published: June 2021

Most studies focusing on neural development and the mechanism of neurodevelopmental disorders are based on animal (rodent or avian) models or post mortem human samples. Thus, the observations derived from these studies cannot be directly translated into mechanistic insights of human neural development and regeneration. Human pluripotent stem cells, which can potentially be differentiated into any cell types of the body, offer a new perspective to overcome this limitation. In the present study, we differentiated induced pluripotent stem cell lines toward the neural lineage. Using a multi‐step, directed differentiation protocol, specific human neural progenitor cells (NPCs) were generated which are committed to produce PROX‐1‐positive granule cells of dentate gyrus of the hippocampus. These NPCs were characterized in terms of morphology, expression of specific cellular markers, growth capacity, and capability of producing mature neurons. Next we investigated the calcium signals in NPCs and PROX1‐positive hippocampal neurons, expressing GCaMP6fast calcium indicator protein or loaded with Fluo4 calcium sensitive dye, in parallel experiments. We demonstrated the functional presence of several receptors, and also the lack of functioning sodium channels in NPCs, and spontaneous Ca2+ gradients in neurons by both methods. We found that spontaneous calcium gradients and specific, ligand induced calcium signals were not significantly altered in the GCaMP6f expressing cells, suggesting the applicability of GCaMP6f for the investigation of fast calcium signals in neural cultures. We conclude that the characteristics of both the spontaneous and ligand‐induced Ca2+‐signals, as well as their pharmacological modulation can be successfully examined in these human model cells by fluorescence imaging.Support or Funding InformationThis study is funded by the National Brain Research Program (NAP) of Hungary (grant numbers: KTIA_NAP_13‐1‐2013‐0001 to LH and KTIA_NAP_13‐2014‐0011 to JR), and the Momentum Program of the Hungarian Academy of Sciences (LP2012‐025 to LH).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Immunomodulatory Effects of Rhesus Monkey Bone Marrow‐derived Mesenchymal Stem Cells in Serum‐free Conditions
Authors: Liu Fei , Li Yuanmin , Zhang Jie , Lu Yanrong , Cheng Jingqiu 
Published: June 2021

BackgroundMesenchymal stem cells (MSCs) has generated tremendous interest for treating various diseases due to the self‐renewal and differentiation. The great majority studies have indicated the immunoregulatory capability of MSCs, however, most researches are conducted based on the fetal bovine serum (FBS), meanwhile, commercial serum free media are not the promising candidates for cell therapy because of the confidentially uncertain composition. We aimed to develop a new serum free medium (SFM) suited for MSCs and evaluate the biological characteristics and the immunomodulatory potential of MSCs.MethodsWe established a complete chemical serum free media to support the proliferation and expansion of rhesus monkey bone marrow‐derived mesenchymal stem cells (rBMSCs), cells harvested through bone marrow aspiration from nine healthy rhesus monkeys. Passage 0 rBMSCs were cultured in serum containing media (SCM), following cell culture performed with serum free media and serum containing media respectively until P3. Growth kinetics, characterization, flow cytometry analysis, and immunosupressive abilities were evaluated compared with SCM. Moreover, RNA sequencing was used to evaluate the differentiated expressed genes (DEGs) related to immunization in two culture conditions.ResultsCompared to SCM, cells cultured in SFM exhibited better biological characteristics in the proliferation and immunnosupressive abilities. Both of cells showed similar immunophenotype for CD29, CD105, HLA‐DR, CD34, CD45. For RNA‐seq, five high correlated DEGs were enriched in immune system, only CXCL8 was down‐regulated (8 fold changes) acting as a pro‐inflammatory chemokine, which indicated that rBMSCs cultured in SFM had lower immunogenicity.ConclusionsrBMSCs are feasible to be expanded safely in SFM and maintain the biological characteristics and immunosuppression functions in vitro. Further, this provides a possible cell culture methods for the clinical cell therapy requiring non‐animal derived demands, and improves the immunoregulatory effects of rBMSCs simultaneously, which could contribute to clinical therapies.Support or Funding InformationThis research was supported by Chengdu Giant Panda Breeding Research Foundation (2016cdspt001). Project name: Rhesus Monkey Mesenchymal Stem Cells Biological Study and Resource Pool EstablishmentThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protective Role of UCP2 against Oxidative Stress through Survival Signaling Pathway in Acute Epilepsy induced by Pilocarpine in Rats
Authors: Dutra Marina RH. , Feliciano Regiane Santos , Santana Eduardo T. , Jacinto Kalil R. , Gouveia Telma R. , Serra Andrey J. , Naffah‐Mazzacoratti Maria G. , Silva Jose A. 
Published: June 2021

Neuroprotection is a desirable process in many human neurological disorders, yet complexes mechanisms involved in this field are not completely understood. The pilocarpine epilepsy model causes potent, seizure‐induced excitotoxicity cell death, and mitochondria impairment is also observed in this model. The present study was designed to verify neuroprotection of UCP2 against hippocampal epilepsy, using Real‐time PCR, immunohistochemistry, Western blot and RNA silencing. Herein, we demonstrated that rats subjected to an animal model of epilepsy induced by pilocarpine had increased levels of UCP2 mRNA expression in the acute phase. Additionally, the brains of rats injected with pilocarpine showed increased pro‐inflammatory makers mRNA expression. Moreover, these rats presented augmented p‐AKT expression only in the acute phase of the protocol, concomitantly with UCP2 mRNA augmentation. Rats treated with pilocarpine also presented an increase in oxidative stress, marked by damage caused by lipid peroxidation, higher levels of protein carbonyl, and a decrease in the levels of antioxidant enzymes, SOD and catalase. On the other hand, when pilocarpine‐treated rats received ASO therapy for UCP2, the number of seizures observed in the acute phase was increased compared to pilocarpine only injected rats. UCP2 silencing were able to worsening the oxidative stress parameters and diminishing p‐AKT expression. In conclusion, our study demonstrated that UCP2 might have neuroprotection after pilocarpine insult in hippocampi of rats.Support or Funding InformationThe authors wish to thank the CNPQ (308067/2014‐2) and FAPESP, PRONEX, Instituto Nacional de Neurociência Translacional, (INNT‐MCT), and CAPES.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Different Effects of Photobiomodulation Therapy on Cultured Human Chondrocytes
Authors: Zhang Junlin , Guffey Stephen , Motts Susan D. , Stroup Kyle J. 
Published: June 2021

IntroductionThe purpose of this study was to investigate the effects of photobiomodulation therapy (PBMT) with different wavelengths and fluences on the cell viability of cultured human chondrocytes. The effects of PBMT on chondrocytes have been studied both in vitro and in vivo. However, outcomes vary among reports due to the application of different light parameters.MethodsNormal human chondrocytes growing in 96 well microplates received either blue (464nm), red (625nm), or infrared (850nm) PBMT at different fluences (1 J/cm2, 3 J/cm2, 24 J/cm2) using Dynatronics Solaris 705 Plus through the probe. A cell proliferation assay kit was used to quantify the number of chondrocytes in PBMT treated and control groups. One way ANOVA and Post Hoc Bonferroni were used to compare the differences among multiple groups using SPSS 21. p<0.05 was considered to be significantly different.ResultsWe found that fluences 3 J/cm2 and 24 J/cm2 of blue PBMT significantly decreased the cell viability while 1 J/cm2 of blue PBMT did not produce significant change. Fluences of 1 J/cm2 and 24 J/cm2 of red PBMT significantly increased the cell viability but 3 J/cm2 of red PBMT did not show such an effect. Surprisingly, none of the fluences tested using infrared PBMT showed any obvious change in cell viability.ConclusionThese findings suggest that different parameters (fluences and wavelengths) of PBMT produce different outcomes with regard to chondrocyte proliferation. These findings contribute to our understanding of the biological responses of chondrocytes when exposed to PBMT and indicate that optimal parameters of PBMT need to be determined to reach desired outcomes.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic Inflammatory Exposure Changes the Phenotype of Human Skeletal Muscle Cells
Authors: McManus Christina , Richardson Chante 
Published: June 2021

Study ObjectivesOsteopathic Manipulative Therapy (OMT) is well documented in treating myofascial pain. OMT, when used appropriately, can reduce acute and chronic pain and improve myofascial function. Chronic pain, skeletal muscle wasting, and skeletal muscle weakness have been associated with inflammatory diseases. Skeletal muscle is a target for OMT and studies have shown that skeletal muscle weakness and wasting has been associated with inflammatory diseases. However, the molecular mechanisms driving these benefits with OMT are not well understood. Our objective is to look at how chronic inflammation, utilizing TNF‐alpha, changes the phenotype of human skeletal muscle cells (hSkM) and what affects stretching, which mimics OMT, improves these phenotypical changes. We hypothesize that chronic inflammation changes the phenotype of the myofascial tissue that can be reversed by OMT.MethodsHuman skeletal muscle cells were grown in skeletal muscle cell growth medium in a 150 mm × 25 mm dish. After reaching confluence, the hSkM cells were split and grown in five T‐75 flasks. Two of these flasks served as controls. The remaining three flasks were exposed to TNF‐alpha in varying concentrations; 0.1, 0.05, and 0.025 microgram/microliters of TNF‐alpha in distilled H2O. hSkM cells were phenotyped by looking at cell shape, cell count, and organization of fluorescent biomarkers.ResultshSkM cells changed shape in response to chronic exposure to pro‐inflammatory factor TNF‐alpha in a dose dependent manner. Compared to control, hSkM cells chronically exposed to TNF‐alpha had cell‐cell contact and organizational changes. TNF‐alpha exposure also decreased cell count in a dose dependent manner.ConclusionsChronic exposure to pro‐inflammatory factor TNF‐alpha changes the hSkM cell phenotype. We predict that cell stretching will reverse these phenotypical changes.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hydrogen Peroxide Causes Iron Dysregulation in C2C12 Skeletal Muscle Cells
Authors: Anderson Jacob G. , Hans Rebekah C. , Symkins Dylan D. , Hancock Chad R. 
Published: June 2021

Diabetes is becoming an increasingly prevalent issue in the United States. Diabetes and insulin resistance is associated with increased levels of circulating iron and signs of iron dysregulation. Unbound iron in cells leads to increased amounts of free radicals causing damage, including insulin resistance. Ongoing research in our lab has shown that insulin resistant rats displayed iron dysregulation in liver tissue, marked by hepatic iron accumulation, with a decreased ability to store the excess iron in ferritin storage proteins. The purpose of this study is to develop a cell model of iron dysregulation to replicate findings in previous studies in order to identify the mechanism by which this iron dysregulation occurs. Using C2C12 mouse skeletal muscle cells, we investigated the effects of hydrogen peroxide on iron regulation, and its role in the development of iron dysregulation. C2C12 cells were treated with 200 μM hydrogen peroxide for 12 hours. Following treatment with hydrogen peroxide, cells were harvested for protein and iron analysis. Protein levels of transferrin receptor (TfR) and ferritin heavy chain (FHC) were measured using western blot analysis. Total iron was measured using a total iron colorimetric assay. Hydrogen peroxide treatment decreased TfR protein levels by 24±16% (p=.03). We then developed a cellular model to reflect the stress caused by elevated circulating iron levels by treating C2C12 cells with varying concentrations (0–100 μM) of FeCl3 for 24 hours. Iron treatment resulted in an increase in total cellular iron in a dose‐dependent manner. As expected, exposure to elevated iron caused a reduction in TfR expression and an increase in FHC in a dose‐dependent manner. Using this high iron model with a 24‐hour treatment of 10 μM FeCl3, we added 200 μM hydrogen peroxide during the last 12 hours of iron treatment to impose conditions of elevated oxidative stress. The combination of iron and hydrogen peroxide resulted in an increase in TfR by 46±9% (p=.05) and a decrease in FHC by 49±13% (p=0.018), showing a complete reversal compared to hydrogen peroxide treatment alone. Total iron increased by 69±6.7% (p=0.0005). Our results show that treatment with hydrogen peroxide in the presence of iron causes an accumulation of iron and a decrease in ferritin expression, potentially leading to an increase of free iron and the production of reactive oxygen species. This represents a condition of iron dysregulation and increased oxidative stress that can contribute to insulin resistance as well as other cellular pathologies.Support or Funding InformationSupported by BYU Mentoring Enhancement GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.