bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of MED28 and Vitamin D on Glycolytic Metabolism and Cell Growth in Human Colorectal Cancer Cells
Authors: Weng Yu‐Ting , Li Chun‐I , Liu Hsiao‐Sheng , Lee Ming‐Fen 
Published: June 2021

Mammalian Mediator Complex has been implicated in human diseases, including cancer and metabolic disorder. MED28, a Mediator subunit, was found highly expressed in several types of cancer. Vitamin D, a secosteroid with hormone‐like features, exhibits multiple biological roles, including calcium homeostasis and cell differentiation. Calcitriol, 1,25‐dihydroxyvitamin D3, and its analogues have been suggested to suppress cancer development through vitamin D receptor (VDR). Our laboratory has previously reported that MED28 mediates Wnt/β‐catenin signaling and glucose metabolism in human colorectal cancer (CRC) cells, and we have also reported that calcitriol inhibits MED28‐mediated cellular events; in this study, we further examine their underlying mechanisms. First, by immunohistochemistry, we observed higher expression levels of MED28 in cancer tissues than those of adjacent normal counterparts in a CRC tissue microarray containing 19 paired tissue samples. Secondly, we detected lower protein expression of MED28, glucose transporter 1 (GLUT1), lactate dehydrogenase (LDHA), and cyclin D1, and less lactate production and higher pH value of the medium when we cultured human CRC cells in low glucose medium as compared with high glucose (25 mM) medium. Thirdly, the employment of siMED28 decreased both glucose uptake and lactate production, accompanied by lower expression levels of GLUT1 and LDHA as well as cyclin D1. Calcitriol inhibited the expression of MED28, and the effects of calcitriol on glucose metabolism and cell growth resembled those of MED28 suppression. In contrast, ectopic expression of MED28 increased glucose uptake as well as the protein expression of GLUT1, LDHA, and cyclin D1. Moreover, MED28 appeared to co‐localize and physically interact with VDR. Taken together, our findings indicated that MED28 upregulated Warburg effect and cell growth, including glucose uptake, lactate production, and the expression levels of GLUT1, LDHA, and cyclin D1, and calcitriol inhibited these events, where the interaction of MED28 and VDR may be responsible for the suppressive effect of calcitriol on MED28.Support or Funding InformationThis work was supported by the grant MOST 106‐2320‐B‐039‐062‐MY3 to M‐F Lee.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Loss of the Retinoblastoma Protein Modulates Glucose Metabolism in Lung Cancer
Authors: Clem Brian , Dougherty Susan , Kruer Traci , He Liqing , Yin Xinmin , Zhang Xiang , Reynolds Lindsey 
Published: June 2021

Dysregulated metabolism is a hallmark of cancer cells and is driven in part by specific genetic alterations in various oncogenes or tumor suppressors. The retinoblastoma protein (Rb) is a tumor suppressor that was initially characterized in regulating cell cycle progression but recent studies have highlighted a functional role for Rb in controlling cellular metabolism. As such, we aim to characterize the metabolic phenotype caused by genetic loss of Rb in a mouse model of lung cancer. We have employed the Kras+/LSLG12D/Rb1lox/lox and Kras+/LSLG12D/Rb1+/+ transgenic model to study the metabolic effects of Rb1 loss in lung cancer. Cre‐exposed Kras+/LSLG12D/Rb1lox/lox mice exhibit increased tumor burden as well as decreased overall survival compared to Kras+/LSLG12D/Rb1+/+ mice. Our tracer studies using bolus dosing of 13C‐glucose revealed an increase in glucose carbon incorporation into select glycolytic intermediates. Consistent with this result, Rb1 depleted tumors expressed higher hexokinase 2 and glucose transporter 1. Additionally, compared to Rb1‐proficient tumors, Rb1 null tumors exhibit decreased one and three carbon labeled aspartate, which is indicative of pyruvate carboxylase (PC) activity. However, Rb1−/− tumors were found to express higher PC compared to Rb1+/+ tumors, suggesting these tumors may utilize an alternative nutrient source for TCA anaplerosis other than glucose. Taken together, these data have identified several metabolic genes under Rb control, and suggest Rb may regulate differential nutrient utilization in lung tumors.Support or Funding InformationNCI RO1CA166327, American Cancer Society RSG 13‐139‐01This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TNF ligand related molecule‐1A inhibits atherosclerosis in apoE‐deficient mice
Authors: Zhao Dan , Yang Xiaoxiao , Chen Yuanli , Duan Yajun , Han Jihong 
Published: June 2021

TNF ligand related molecule‐1A (TL1A) is one of vascular endothelial growth inhibitors. The regulation of neovascularization by TL1A indicates that it may play a vital role in the development of atherosclerosis. However, the exact effect of TL1A on the development of atherosclerosis and the underlying mechanisms, such as the functions of TL1A on foam cell formation and vascular smooth muscle cell (VSMC) phenotype, remain unknown. In this study, we induced atherosclerosis in apoE deficient (apoE−/−) mice by feeding the animals a high‐fat diet (HFD) while intraperitoneally injecting them recombinant TL1A twice a week, for 12 weeks. After treatment, we collected mouse aorta, serum and liver samples to determine the effect of TL1A on atherosclerotic lesions and fatty liver, and expression of the related molecules. The results of Oil red O, VVG and Alizarin red S staining show that administration of TL1A led to a significant reduction of lesions while enhancing plaque stability. The inhibition of lesions by TL1A was attributed to its effect on formation of foam cells derived from SMCs, but not from macrophages. Corresponding, TL1A activated expression of ABCA1 and ABCG1 in SMCs, but not in macrophages. Both the in vivo and in vitro results showed that TL1A inhibited the transformation of SMCs from contractile phenotype into synthetic one by inducing expression of contractile markers (α smooth muscle actin or αSMA) and inhibiting expression of synthetic marker (osteopontin, OPN). In addition, we determined that TL1A treatment decreased liver triglyceride contents by upregulation of HSL and ATGL expression. Taken together, our work shows that TL1A can inhibit the development of atherosclerosis by regulating SMC/foam cell formation and SMC phenotypes. Our study may also provide a new strategy for atherosclerosis treatment.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
para‐substituted methcathinones as selective and unselective inhibitors of human dopamine and serotonin transporters
Authors: Niello Marco , Cintulova Daniela , Hellsberg Eva , Jäntsch Kathrin , Holy Marion , Kudlacek Oliver , Cozzi Nicholas , Baumann Michael H , Mihovilovic Marko , Ecker Gerhard , Sitte Harald 
Published: June 2021

Methcathinone (MCAT) is a compound belonging to the class of cathinones which targets monoamine transporters including DAT and SERT. Despite the importance of DAT and SERT as drug targets in several neurological disorders, the key factors underlying the selectivity profiles of their inhibitors are still poorly understood. Recent findings from rat synaptosomes suggested that increasing the volume of the para‐substituent of MCAT results in a swap of the selectivity between human DAT and SERT (1). Docking studies hint towards Ser149 in DAT and Ala169 in SERT as key residues involved in the difference of activity between DAT and SERT (2). The aims of the present biochemical and pharmacological study are to understand (i) which chemical properties (e.g. volume, polarity, or lipophilicity) of the para‐substituent influence the selectivity profile of MCAT between DAT and SERT, and (ii) whether Ser149 in DAT and Ala169 in SERT can be experimentally verified as key residues. Hence, we combined in silico‐driven synthesis, mutagenesis, and radio‐tracer flux assays in HEK293 cells expressing the human DAT and SERT wild‐type and respective mutants. We found that only MCAT and the para‐CF3‐MCAT showed high selectivity, 200‐fold for DAT/SERT ‐ and 25‐fold for SERT/DAT, respectively. This suggests that the high selectivity achieved is determined rather by specific features of these compounds than by the volume of the para‐substituent. Accordingly, we were not able to find any correlation between the selectivity profile of the tested 4MCATs with either volume, polarity, or lipophilicity parameters. Next, based on our results, we tested the hypothesis that the two amino acids in SERT and DAT do not suffice to explain the selectivity between SERT and DAT. We have tested the para‐substituted methcathinones in the swapping mutations DAT Ser149Ala and SERT Ala169Ser and in line with our hypothesis, these mutations did not revert the selectivity profile found in the wild‐type transporters. Our findings rather suggest that the SERT selectivity achieved by the introduction of the CF3‐group in the para‐position of MCAT is not dependent on the volume of the para‐substituent but on specific chemical features of the fluorine atoms which may influence the on and off rate of the MCAT molecule on DAT and SERT.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Therapeutic effects of melatonergic networks on propionic acid‐mediated autism‐like rat model: Sex difference based study
Authors: Kim Joo‐Heon , Choi Jeonghyun , Hong Yonggeun 
Published: June 2021

Autism spectrum disorder (ASD) is characterized not only by altered neurobehavioral symptoms including repetitive and restricted behaviors, defects in social interactions, intestinal malfunctions such as constipation, and sleep disorders but also pathological brain damages. As the cause of this disorder is not clear, early diagnosis is limited and it may remain permanently disabled. Therefore, this study aims to explain the therapeutic potential of melatonergic network‐related materials in propionic acid‐mediated ASD animal models. Especially, to identify genderdependent changes, male and female 3‐week‐old Sprague‐Dawley rats were randomly divided and used. They were divided into control, propionic acid‐mediated ASD group, ASD with melatonin treatment group, and ASD with fermented milk treatment group. To verify the therapeutic effects of melatonergic‐related materials, intestine length, sleep quality and neurobehavioral tests were performed. ASD animals showed decreased intestine length compared to control. Interestingly, exogenous melatonin and fermented milk treatment to ASD animals restored intestine length in both sexes. However, no difference was found in the intestine length of both genders. Also, melatonin and fermented milk administration to ASD group showed restored sleep quality regardless of sexes. Additionally, ASD animals showed abnormal neurobehaviors compared to control animals, and this abnormality is thought to be recovered by exogenous melatonin and fermented milk in both male and female animals. Accordingly, melatonergic network may exert its potential as a therapeutic agent in ASD.Support or Funding InformationNRF‐2017R1A2A2A01067169, KGM4611821, 2016 Creative Research Program of Inje University, Republic of KoreaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biomarkers of Neurodegeneration in Goldfish Exposed to Nitrite
Authors: Valdez‐Oranday Diego Francisco , Martinez Melody , Huertas Mar 
Published: June 2021

Goldfish have been used in numerous studies involving behavior and toxicology, however, their viability as an animal model to study neuropathies remains to be elucidated. We have previously discovered that the nitrite ion (NO2−) acts as an olfactory disruptor in goldfish. We hypothesize that this disruption is due to neurotoxic effects of NO2− in olfactory receptors and the brain. In this study, goldfish were subjected to a subacute, sublethal 1.0 mM sodium nitrite treatment, and had their brains and olfactory tract dissected. The tissues were mounted in paraffin blocks for sectioning and visualized with hematoxylin and eosin (H&E), Alcian blue, terminal deoxynucleotidyl transferase dUTP nick‐end labeling (TUNEL), caspase immunohistochemistry, and Congo red β‐amyloid staining. We discovered that olfactory tissue from treatment fish displayed signs of hyperplasia and hypertrophy when compared to the olfactory tissue from control fish. Furthermore, signs of increased apoptotic processes, in the form of blebbing, were observed in treatment fish. Alcian blue staining for mucosal cells demonstrated that the microvilli lining the epithelia had lost their brush‐like morphology and were covered by a thick layer of mucus in treatment fish. TUNEL, caspase immunohistochemistry, and β‐amyloid stains are currently being analyzed. Our results indicate that NO2− acts as a neurotoxin in subacute, sublethal doses. Because of the similarities in olfactory processing between goldfish and other vertebrates, it is possible that NO2− may also play a similar role in other aquatic vertebrates. The general availability and easy handling of goldfish make it an ideal candidate to study neuropathies.Support or Funding InformationTexas State University start‐up funding for Dr. Mar HuertasThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Natural Rubber Latex as Stem Cell Scaffold for Muscle Regeneration
Authors: Floriano Juliana Ferreira , de Oliveira Graeff Carlos Frederico , Barbosa Angélica Mércia Pascon , Marcondes João Paulo , Pinto Taís Silva , Zambuzzi Willian Fernando , Rudge Marilza Vieira Cunha 
Published: June 2021

Natural rubber latex (NRL) is a product extracted from rubber trees. The most common source of NRL is Hevea brasiliensis, NRL is a mixing colloidal rubber particle stabilized by a thin layer of phospholipids and proteins. The NRL has several features that make it an excellent scaffold and biomaterial to promote the growth and repair of tissues, due to their characteristics, like biocompatibility, angiogenic potential, flexibility, mechanical stability, surface porosity, permeability and low cost. The present study aimed at assessing the potential NRL use as stem cell scaffold, for the future use in muscle regeneration and cell therapy. The NRL membranes were produced with latex collected from RRIM 600 clone, the NRL was stored at a low temperature to prevent spontaneous coagulation and was centrifuged for 5 min at 1,500 rpm to remove impurities gathered in the collection process. The membranes were prepared by depositing a thin layer of NRL covering the bottom of a polystyrene petri dish measuring 60 mm in diameter and carried to a oven, where it was dried at 50 °C. The study was approved by Botucatu Medical School ethics committee on the use of animals (CEUA number 1234/2017). The mesenchymal stem cells were isolated from the bone marrow of the femur of young rats and cultured until the fourth passage. After which the cells were characterized by immunocytochemistry and immunophenotyping techniques, cell adhesion on the surface of the NRL membrane after 7 days of sowing the cells was assessed by scanning electron microscopy (SEM), confocal microscopy, zymography techniques for the evaluation of metalloproteinases 2 and 9, electrophoresis and protein expression for integrins. Our partial results showed that 98% of the cells expressed CD90, besides adhesion to plastic and the ability to differentiate and formation of fibroblast colonies, according to the methods indicated by the International Society for Stem Cell. After being seeding the surface of the NRL membranes it was verified high cell viability, excellent cell adhesion high cell proliferation rate, demonstrating the great potential NRL as stem cell scaffold.Support or Funding InformationThis project is funded by São Paulo Research Foundation – FAPESP (number grant 2016/01743‐5)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 effects of hyperglycemia on functionality of Rectus abdominis muscle of pregnant women
Authors: REYES DAVID RAFAEL ABREU , Barbosa Angélica Mércia Pascon , Floriano Juliana , Quiroz Sofia Mércia Vega , Enriquez Eusebio Mércia , Marcondes Joao Paulo , Okoshi Marina Mércia Politi , Rudge Marilza Mércia Vieira Cunha , Calderon Iracema Mércia , Raghavendra Hallur Mércia 
Published: June 2021

IntroductionGestational diabetes mellitus (GDM) is an important risk factor for urinary incontinence (UI) two‐year post caesarean section (CS). Previous studies using rectus abdominis muscle (RAM) of pregnant rats with severe and moderate diabetes confirmed diabetic myopathy as a risk factor to UI. But the pathophysiological mechanisms of UI are not completely understood. Indeed, hyperglycemia can damage skeletal muscle with atrophy due to neuropathy or direct effect on muscle cell metabolism. In this context, this study evaluated for the first time the effects of hyperglycemia on the biomechanics of the RAM of pregnant women using Miography − an innovative technique, ex‐vivo, sensitive, that evaluates the function of the myocardial fiber through a controlled electrical stimulation.ObjectiveTo identify the possible deleterious effects of hyperglycemia on RAM functionality (ex vivo contractility) in pregnant women with and without GDM.MethodologyRAM of four primipara women with and without IU (three normoglycemic and one hyperglycemic) were collected during CS. The surgeon collected 1 cm in length of RAM and placed it in Krebs solution at 4°C. The changes in isometric tensile strength were recorded using LabChart software (LabChart 7 for Windows, AD Instruments) coupled to PowerLab (The PowerLab Data Acquisition System, AD Instruments) software for capturing and storing muscular responses. Muscle mechanical responses were expressed as a percentage of maximal contraction induced by electrical stimulation (Grass Model S48). We performed a multiple regression analysis adjusted to confounders using SPSS v21.o software. p < 0.05 was considered as statistically significant.Initial resultsWe obtained results of the RAM function after electrical stimulation during one hour of the four women. We noticed that the RAM of pregnant women does not sustain the electrical response during one hour of the experiment. We have not enough data to compare both groups, but I confirmed that the myography is effective to evaluate the RAM functionality.ConclusionThese initial results confirmed myography as a viable technique to analyze the functionally of the RAM in pregnant women, in a controlled way. The effects of hyperglycemia on the functionality of the RAM were identified.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Topical Application of Recombinant Human MG53 Protein Facilitates Healing of Chronic Wounds in Diabetic Mice
Authors: Zhou Xin , Li Haichang , Zhou Xinyu , Tan Tao , Xu Xuehong , Ma Jianjie 
Published: June 2021

Wound care is a major challenge to public health and the burden for wound treatment is growing due to the aging population and the sharp rise in the incidence of diabetes and obesity, diseases that contribute to poor wound healing. While multiple factors may contribute to complications associated with wound healing, compromised tissue repair represents an underlying cause for non‐healing wounds in diabetes. We previously identified a novel TRIM family protein, named MG53, as an essential component of the cell membrane repair machinery. Here we tested the therapeutic value of using recombinant human MG53 (rhMG53) protein to treat chronic wounds. We developed a prototype formulation of rhMG53 in a topical cream that remains stable for long‐term storage at 4°C and is suitable for wound healing applications. We developed a mouse model of chronic excisional dermal wound using the db/db mice. Under control conditions, dermal wounds in the db/db mice would not heal until day 21. Treatment with topical cream containing 1.5 and 10 mg/oz of rhMG53 accelerated wound healing. Application of the topical cream without rhMG53 was ineffective. Wounds treated with 10 mg/oz rhMG53 cream healed nearly completely by day 12. Histological analyses revealed remarkable improvement of hair follicle structure at the dermal wound site in db/db mice that received treatment with rhMG53; the healing site that received cream alone showed defective hair follicles. We have previously demonstrated that genetic ablation of MG53 led to defective hair follicle structure in the mg53−/− mice. Our findings suggest that MG53 could improve chronic wound healing in diabetes by enhancing the hair follicle structure and the regenerative capacity of the skin.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Blockade of the AT1 receptor and silencing of CD44 gene expression are associated with inhibition of angiotensin II‐activated cardiac fibroblasts
Authors: Bai Feng , Yang Guang‐Zhao , Wang Ning‐Ping , Zhao Zhi‐Qing 
Published: June 2021

ObjectiveWe have previously demonstrated that the stimulation of fibroblasts by angiotensin II (Ang II) contributes to cardiac fibrosis via transforming growth factor beta (TGF‐β1)‐mediated collagen production. The cellular/molecular mechanism responsible for fibroblast activation, however, is not well understood. CD44, a cell‐surface glycoprotein, is associated with cell‐cell interactions, cell adhesion and migration. This study examines the roles of Ang II AT1 receptor and CD44 gene expression in the activation of cardiac fibroblasts.MethodsCardiac fibroblasts were isolated from the neonatal rat hearts, the assays of cell viability and migration were selected to demonstrate the activation of fibroblasts, and silencing of CD44 gene expression on fibroblasts was conducted with small interfering RNA (siRNA).ResultsImmunofluorescence staining with vimentin and discoid in domain receptor‐2 revealed the purity of isolated fibroblasts. Ang □ significantly reduced the cell viability and increased the cell migration in a dose‐dependent manner with a maximal stimulation as seen at 50 nM. In coincidence with fibroblast activation, mRNA and protein level of CD44 were up‐regulated and the protein levels of TGF‐β1, Smad2/4 and collagen I were significantly increased (all p<0.05 vs. unstimulated cells). Addition of Ang II AT1 receptor blocker, telmisartan into the cells abolished Ang II‐elicited stimulating effects on these parameters measured (all p<0.05 vs. Ang II activated cells). Consistent with data showing that there is a linear correlation between CD44 and TGF‐β1 as demonstrated by Pearson correlation analysis (r=0.955, p<0.01), gene transfection of fibroblasts with Ad‐CD44 siRNA as identified by mRNA and protein levels of CD44 demonstrated a significant attenuation in the production of collagen I, suggesting a role of CD44 in the activation of fibroblasts.ConclusionsThese results indicate that cardiac fibroblasts, in response to Ang II stimulation, express high levels of the AT1 receptor and CD44, which reduce the cell viability and facilitate cell migration. Such a signaling mechanism could be crucial for leading to the deposition of collagen and the production of cardiac fibrosis.Support or Funding InformationThis study was supported in part by grants from the Mercer University School of Medicine, the Medcen Community Health Foundation, Georgia and the National Natural Science Foundation of China (81470436).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.