bluuebunny/crossref_metadata_2025_split huggingface dataset:
The FOXM1 Inhibitor RCM‐1 Decreases Carcinogenesis and Nuclear β‐catenin
Authors: Rama Nihar , Shukla Samriddhi , Milewski David , Kalin Tanya 
Published: June 2021

FOXM1, a member of the Forkhead box (FOX) family of transcription factors, has previously been shown to play a critical role in carcinogenesis by inducing cellular proliferation in multiple types of cancer when abnormally upregulated. Our lab previously identified a small molecule compound, Robert Costa Memorial drug‐1 (RCM‐1), through high‐throughput screening as a FOXM1 antagonist in vitro and in an in vivo mouse model of allergen‐mediated lung inflammation. Here, we examined antitumor activities of RCM‐1 using tumor models. Confocal imaging of RCM‐1‐treated tumor cells indicated that reduced cellular proliferation was concordant with inhibition of FOXM1 nuclear localization in these cells. RCM‐1 reduced the incidence and growth of tumor cell colonies in the colony formation assay. Migration assays in vitro demonstrated the ability of RCM‐1 to inhibit migration of tumor cells. In animal models, RCM‐1 treatment inhibited growth of mouse rhabdomyosarcoma Rd76‐9 and human lung adenocarcinoma H2122. RCM‐1 decreased FOXM1 protein levels in treated tumors, reduced tumor cell proliferation and increased tumor cell apoptosis in vivo as detected by immunohistochemical staining. Western blot analysis of cultured tumor cells showed that RCM‐1 decreased protein levels and nuclear localization of β‐catenin, an important constituent of the canonical Wnt signaling pathway, and inhibited protein‐protein interaction between β‐catenin and FOXM1. Altogether, our study provides important evidence of antitumor activity of the small molecule compound RCM‐1, suggesting that RCM‐1 has promising potential as a cancer therapy in the future.Support or Funding InformationPeter K. Lauf Award, Ohio Physiological Society; NIH funding awarded to TK.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Histological Analysis of Lysophosphatidic Acid Receptor 3 Deficient Zebrafish
Authors: Chen Wei‐Min , Chang Ya‐Chi , Lin Yu‐Nung , Chiang Jui‐Chung , Lee Hsinyu 
Published: June 2021

Lysophosphatidic acid (LPA) is a small bioactive phospholipid enriched in serum and exerts diverse cellular functions through activation of multiple G Protein‐coupled receptors, LPA1‐LPA6. Our previous studies demonstrated that lysophosphatidic acid receptor 3 (LPA3) is critical for megakaryopoiesis and erythropoiesis in zebrafish. However, the roles of LPA3 during the development of other organs of zebrafish remain unclear. Here, we established a zebrafish strain with targeted disruption of lysophosphatidic acid receptor 3 (Lpar3) gene by Transcription Activator‐like Effector Nuclease (TALEN). Paraffin‐section and hematoxylin‐eosin staining were performed for histological analysis to clarify the effects of LPA3 in different organs of adult zebrafish. Histological analysis results showed a shrinking of hematopoietic tissue in kidney marrow of LPA3 deficient fish, implies essential roles of LPA3 in hematopoiesis. In addition, bile duct hyperplasia and mature spermatids loss were also observed in LPA3 deficient fish. Nevertheless, no obvious morphological differences in eyes, thymus, heart, and spleen were observed. Our observation provides tissues profiles of LPA3−/− animal and possible application of LPA3 deficient zebrafish as a disease model in the future.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 Function in Dopamine Receptor‐3 (D3R) Attenuates Left Ventricular Cardiac Fibroblast Migration and Proliferation In Vitro
Authors: Kisling Andrew , Zakari Madaniah O , Alsahly Musaad B , Melit‐Thomas Deepthy C , Clemens Stefan , Lust Robert M , Katwa Laxmansa C 
Published: June 2021

Dopamine is a neurotransmitter heavily involved in neural pathways regulating the reward response, body movement, mood, and cognitive function. As such, much of the research on dopamine and its five receptor subtypes has been conducted to observe the in vivo function of each receptor subtype in neurological disorders. However, recent studies by colleagues from our institution have suggested that D3Rs may play a role in cardiac‐related aging, as 2‐month old D3 receptor knock‐out (D3KO) mice show age‐related changes in cardiac function similar to 2‐year old wild‐type (WT) mice. Thus, it would be interesting to understand the role of D3R expression in LV cardiac fibroblast and its function. We isolated, cultured and characterized cardiac fibroblasts from the left ventricles (LV) of WT (23‐weeks old) and D3KO (5 and 31‐week‐old) mice. Immunohistochemistry was performed to determine the expression of D3R in LV cardiac fibroblasts, and cell culture experiments were performed to examine possible changes in WT and D3KO cell migration and proliferation. Proliferation was measured via total cell count over time points of 6, 12, 24, and 36 hours, while migration in response to scratch wound was examined as percent change in area at 3, 6, 12, 24 hours, and every 12 hours thereafter up to 86 hours. Staining with antibodies for D3R successfully showed expression of this receptor in WT fibroblasts, while cell culture experiments showed reduced proliferation and migration of D3KO fibroblasts compared to WT. Migration of D3KO fibroblasts from 5‐week old mice were significantly different (*p<0.05) in response to an artificial wound when compared to 23‐week‐old mice. Additionally, upon isolation of LV cardiac fibroblasts from 5‐week‐old and 31‐week‐old D3KO and 23‐week‐old WT mice, cell volume from D3KO fibroblast isolation was markedly reduced compared to WT isolation under identical conditions (5‐fold and 10‐fold decrease in 5‐week and 31‐week D3KO, respectively). This data suggests that D3Rs play a role in LV cardiac fibroblast migration and proliferation and support previous observations of age‐related changes in cardiac function of D3KO mice.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Circulating muscle‐enriched microRNA expression correlates with myogenic gene expression in chronic alcohol administered SIV‐infected macaques
Authors: Simon Liz , Molina Patricia 
Published: June 2021

BackgroundSkeletal muscle enriched microRNAs, myomiRs, regulate gene expression critical for myogenesis, metabolism and functional muscle mass. Previously we demonstrated that chronic alcohol and simian immunodeficiency virus infection decreased myogenic differentiation capacity of myoblasts, and this was associated with a decrease in myomiR expression. Evidence suggest that exosomes carrying myomiRs target myocytes and modulate their physiology and function. The aim of this study was to determine whether total and exosomal circulating‐myomiR expression correlates to myogenic gene expression and myoblast differentiation potential.MethodsGene and microRNA expression was determined in skeletal muscle tissue from simian immunodeficiency virus (SIV) infected and chronic binge alcohol (CBA) administered rhesus macaques with and without antiretroviral therapy (ART) (VEH/SIV/ART−, VEH/SIV/ART+, CBA/SIV/ART, CBA/SIV/ART+). Plasma exosomes were isolated using the total exosome isolation kit based on the principle that by sequestering water molecules, less‐soluble vesicles are forced out and is isolated by low‐speed centrifugation. Total circulating myomiR expression was determined in plasma using taqman miRNA primers and cel‐238 used as the spike‐in control.ResultsCBA decreased circulating total and exosomal miR‐206 expression. Expression of exosomal miR‐206 was positively correlated with miR‐206 expression in the muscle. Circulating and total exosomal miR‐206 also correlated with the myotube density of myoblasts and was negatively correlated with histone deacetylase 4 expression in the different treatment groups.ConclusionsChronic alcohol decreases circulating myomiR expression in myoblasts isolated from SIV‐infected animals. Whether circulating myomiR expression can be used as a noninvasive biomarker to predict dysfunctional muscle mass will be determined in a cohort of people living with HIV with alcohol use disorder has to be determined.Support or Funding InformationK01AA024494 and AA09803 NIH/NIAAAThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SOX18 Variant E137K Reduces Transcriptional Activity of SOX Responsive Promoters in Cotransfected Epithelia
Authors: Sirpilla Olivia , Prokop Jeremy W. , Qutob Dinah , Freeland Thomas , Underwood Adam C. 
Published: June 2021

SOX (SRY‐related box) proteins are important regulators of cellular differentiation during both embryonic development and throughout life affecting overall systemic health. Mutations within SOX genes have been found to cause many forms of cancer, cardiovascular disease, neurological abnormalities and sex reversal. Each SOX gene contains a highly conserved High‐Mobility Group‐box (HMG box) binding domain that changes transcriptional activity of target promoters by bringing distant regulatory components closer to the initiation complex. After deep sequence analysis of all 20 SOX genes, we identified a novel mutation within the SOX18 HMG‐box that exchanges glutamic acid (E) for lysine (K) at amino acid 137 (rs201931544). This mutation is present in the genome of ~0.82% of the Latino population, yet the biological implications of the E137K variant are not known. To address this, we used a luciferase‐based reporter system to assess the ability of the wildtype verses mutant form of SOX18 to activate gene transcription. First, primers were designed to amplify and clone SOX18 into the pHTCHaloTag® CMV‐neo vector (Promega). Following sequence validation, PCR was used to introduce the mutation followed by cloning of the pHTCHalo/SOX18‐E137K effector construct. Constructs were transfected into two epithelial cell lines (HeLa and A375) and activity of a luciferase reporter containing an SOX inducible synthetic promoter was measured (pGL3/AR600). Cells were transiently cotransfected with effector constructs (either Halo/SOX18, Halo/SOX18‐E137K, or empty Halo vector), the pGL3 firefly luciferase reporter as well as a control vector. Transcriptional activity facilitated by each effector was measured for luciferase activity. Results show markedly reduced luminescence (1.3 fold) in cells co‐transfected with E137K relative to native SOX18. This is a significant finding as it validates the functional implication of this mutation. Future studies focusing on additional variants found within the third alpha helix of SOX18 are currently underway.Support or Funding InformationSupport for this study was from NIH‐K01ES025435 (to JWP) and Walsh UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Role for ABCC4 in Regulating Pigment Granule Aggregation in Mice
Authors: Mireles Thomas , Ibarra Dora Evelyn , Wiebe Emely Reimer , Pattillo Amanda M , Rodriguez Cody J , Schuetz John D , García Dana M 
Published: June 2021

ABCC4 is member of the family of ATP‐binding cassette proteins that specialize in transporting a variety of molecules out of the cell. ABCC4 exports signaling molecules present in the retina, including cyclic adenosine monophosphate (cyclic AMP). In fish, cyclic AMP regulates processes in the retinal pigment epithelium (RPE), such as dark‐adaptive pigment granule aggregation. In mammals, cyclic AMP accumulates in photoreceptors in the dark. If cyclic AMP were exported from the photoreceptors into the sub‐retinal space, it could then be available for uptake by the RPE, in which it could activate processes that drive pigment granule aggregation. We hypothesize that ABCC4 in photoreceptors mediates cAMP‐export and is needed for pigment granule migration in mice. To investigate ABCC4's involvement in pigment granule aggregation in mice, pigment granule density was measured along the basal microvilli in the RPE in both wildtype (N=3) and Abcc4 knockout mice (N=3). Pigment granule densities measured in the RPE of wild type, dark‐adapted mice differed significantly from those measured in wildtype, light‐adapted mice (p‐value = 0.02). In contrast, pigment granule density was not statistically significantly different between Abcc4 knockout mice that had been dark‐adapted or light‐adapted (p‐value = 0.13). These results support the hypothesis that ABCC4 is involved in the process of pigment granule migration in mice and are consistent with a novel role for cyclic AMP as a cell‐to cell signaling molecule in the retina.Support or Funding InformationThis project was supported by Texas State University in the form of an undergraduate research fellowship to AMP and funds from the Office of the Provost awarded to DMG in recognition of her service as Presidential Fellow. DEI received salary support from HSI STEM Program (84.031c) Award #P031C160035.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
NR4A2 Inhibits ERK‐Mediated Protein Synthesis and Cell Growth in Response to Beta‐Adrenergic Stimulation in Cardiomyocytes
Authors: Ashraf Sadia , Hegazy Yassmin , Harmancey Romain 
Published: June 2021

Sustained elevation of sympathetic activity is an important contributor to pathological cardiac hypertrophy, ventricular arrhythmias, and left ventricular contractile dysfunction in chronic heart failure. The orphan nuclear receptor NR4A2 is an immediate early response gene activated in the heart under beta‐adrenergic stimulation. The goal of this study was to identify the transcriptional remodeling events induced by NR4A2 expression in cardiomyocytes, and their impact on the physiological response of those cells to sustained beta‐adrenergic stimulation. Treatment of adult rat ventricular myocytes (ARVMs) with isoproterenol induced a rapid (< 4 hours) but transient (< 24 hours) increase in NR4A2 expression levels that was accompanied by increased nuclear localization of the transcription factor. Adenovirus‐mediated overexpression of NR4A2 modulated the expression of genes linked to adrenoceptor signaling, calcium signaling, cell growth and proliferation, and counteracted the increase in protein synthesis rate and cell surface area mediated by chronic isoproterenol stimulation. In consistence with those findings, NR4A2 overexpression also blocked the phosphorylative activation of ERK1/2, Akt, and of their downstream effector in protein synthesis p70S6K. Prominent among the transcriptional changes induced by NR4A2 was the > 7‐fold up‐regulation of the dual‐specificity phosphatases DUSP2 and DUSP14, two known inhibitors of ERK1/2. Pre‐treatment of NR4A2‐overexpressing cardiomyocytes with the DUSPs inhibitor BCI prevented the inhibition of ERK1/2 and p70S6K following isoproterenol stimulation. In conclusion, our results suggest that NR4A2 acts as a novel negative feedback regulator of the beta‐adrenergic receptor‐mediated growth response in cardiomyocytes, and this at least partly through DUSP‐mediated inhibition of ERK1/2 signaling.Support or Funding InformationThis work was supported by NIH Grants R01HL136438, R00HL112952, P01HL051971, and P20GM104357.This 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 Maternal High Fructose and Offspring High Fat Intake on Programmed Hypertension in Young Adult: the Role of Nutrient Sensing Signaling in the Brain
Authors: Chao Yung‐Mei , Chan Julie Y.H. 
Published: June 2021

There is now substantial epidemiological evidence suggesting the origin of susceptibility for metabolic syndromes (MetS) and associated cardiovascular dysfunctions in adulthood can be traced back to the early life. We have previously demonstrated that maternal high fructose diet (HFD) induces programmed hypertension in adult offspring; however, the cellular and molecular signaling contributing to the developmental programming of adult hypertension is not fully understood. There are a number of nutrient sensing pathways, including silent information regulator transcript (SIRT), AMP‐activated protein kinase (AMPK), and phosphatidylinositol 3‐kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) located in the central nervous system and are responsible for the detection of sugars, amino acids, lipids and the surrogate metabolite level for the maintenance of metabolic homeostasis. Perturbation of these signaling pathways could lead to MetS and the associated cardiovascular complications. This study was designed to investigate the role of these nutrient sensing pathways in the brain on developmental programming of hypertension in offspring to maternal HFD, and the influence of high fat diet (HFatD) in offspring to these nutrient sensing pathways. Female Sprague‐Dawley rats were fed with 60% HFD or normal diet (ND) during gestation and lactation periods. After weaning both HFD and ND offspring were subjected to HFatD or ND from the age of 3 to 12 weeks. Blood pressure was monitored under conscious condition via tail‐cuff method from week 6 to 12. Expression of nutrient sensing signals in the forebrain hypothalamus was examined by Western blot analysis. Tissue oxidative stress was quantified by electron spin resonance spectroscopy. Maternal HFD induced programmed MetS and hypertension that became significant at age of 9 weeks and maintained in young adult at age of 12 weeks. Changes in both metabolic and blood pressure phenotypes were appreciably enhanced in HFD offspring with additional HFatD intake after weaning. Protein expressions of SIRT1, phosphorylated AMPK (p‐AMPK), and p‐Akt in the hypothalamus were downregulated in the HFD offspring, which were further suppressed in offspring fed with HFatD. These molecular changes were associated with tissue oxidative stress in the hypothalamus.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 fermented milks on physiological and neurobehavioral changes in male Sprague Dawley rats
Authors: Choi Jeonghyun , Kim Joo‐Heon , Ham Jun‐Sang , Hong Yonggeun 
Published: June 2021

Melatonin is a neurohormone secreted by pineal gland and shows pluripotent functions such as anti‐inflammation, anti‐oxidation, and neuronal cell survival. The major function of melatonin is to control circadian rhythm, from awakening to sleep. Recently, many useful items emitting bright light at night time including light emitting diode (LED) and smartphone, have been emerged. They give convenience to human life, however, can cause various diseases including sleep deprivation, cancer, and obesity. Sleep disturbance by light exposure at night can lead to anxiety‐like behavior, depression, and decreased cognitive function. Therefore, this study aims to identify the effects of three types of fermented milks (FM); C, T1, T2, on sleep and neurobehaviors. Experimental animals were randomly divided into four groups; Cont (control croup), C (treatment with type C FM), T1 (treatment with type T1 FM), and T2 (treatment with type T2 FM). To determine the intestinal activation, the length from cecum to colon was measured. In T2 group, cecum/colon length was significantly increased compared to the length of other groups. To evaluate the sleep quality, electroencephalogram (EEG) was measured through intracranial electrode implantation surgery. C, T1, and T2 FM treated groups showed increased activation of δ‐wave than Cont group. Also, neurobehavioral changes were positively improved in T2 group compared to other groups. In addition, melatonin concentration containing in C, T1, and T2 was 56.0 pg/g, 77.6 pg/g, and 504.4 pg/g, respectively. Therefore, melatonin in fermented milk might have positive effects on gut‐brain bidirectional communication.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.
The human primordial microbiome hosts complex bacterial and fungal interkingdom ecologies that vary with gestational age
Authors: Willis Kent A , Purvis John H , Gomes Charles K , Talati Ajay J , Pierre Joseph F 
Published: June 2021

Commensal microbial communities play important roles in human development. While microorganisms appear to be predominately transmitted from mother to child, the timing of transmission remains controversial. Meconium, the first stool passed after birth, was traditionally considered sterile because it originated from a sterile uterine environment. Following advances in next‐generation sequencing (NGS) techniques, more recent evidence suggests bacterial communities may exist in the gut prior to birth, however, the presence of primordial fungal communities has not been described. Here, we demonstrate the presence of multi‐kingdom microbial communities in meconium immediately following birth based on NGS. To describe the ecology of the primordial microbiome, we prospectively enrolled premature (gestational age < 32 weeks') very low birthweight (VLBW < 1500 g) and full term (FT, gestational age 37–41 weeks') newborns and collected the initial meconium. We examined the interkingdom community structure using 16S and ITS rDNA‐based amplicon sequencing followed by culture‐based techniques. We prospectively enrolled 57 VLBW and 35 FT newborns and collected a stool sample at < 48h of life from 34 infants in each group. Bacteria were found in meconium of 90% of VLBW and 82% of FT samples (χ2P=0.325) and fungi in 93% of VLBW and 62% of FT samples (χ2P=0.003). The bacterial and fungal communities differed significantly between VLBW and FT (bacteria: PERMANOVA P=0.006, fungi: PERMANOVA P=0.013). Fungal communities of VLBWs were dominated by Ascomycota, including Candida, while FT meconium contained greater Basidiomycota and taxonomic diversity. Bacterial communities in VLBW meconium were also less diverse and contained on average a third as many taxa compared with FT. These results demonstrate human meconium contains complex interkingdom microbial communities that develop with gestational age, suggesting host‐microbe interkingdom relationships may exist in the uterine environment. Intriguingly, fungi are 50% more commonly observed in premature meconium than in full term neonates which could suggest a pathological association with preterm birth.Support or Funding InformationThe Marshall Klaus Perinatal Research Award from the American Academy of PediatricsThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.