bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhanced Chemotactic Response of MDA‐MB‐231 Breast Cancer Cells Towards Stable Gradients of EGF
Authors: Schwarz Jan , Kuttenberger Verena , Baumann Nina , Horn Elias , Zantl Roman 
Published: June 2021

Chemotactic cell migration is a basic mechanism underlying cancer cell invasion and hence metastasis. In this regard, growth factors such as epidermal growth factor (EGF) have been identified as guidance cues. Using ibidi's μ‐Slide Chemotaxis we confirm the growth factor EGF as potent guidance cue for directional MDA‐MB‐231 migration in a physiological 3D environment. Moreover, we find enhanced sensitivity towards gradients of EGF in serum free media supplemented with defined protein composition (UltraCULTURETM). Combining defined, physiological migration conditions and enhanced sensitivity allowed us to quantify EGF dependent MDA‐MB‐231 chemotaxis in a detailed manor. MDA‐MB‐231 cells showed increased directional and kinetic response towards temporal and stable spatial gradients of EGF. While EGF guided directional migration was highly concentration dependent with a concentration optimum of 10 ng/mL, we found the chemokinetic effect induced by the presence of EGF being concentration independent. Those results suggest differential response of the migration and signal interpretation machinery of MDA‐MB‐231 cells. Both, blocking the ligand binding domain of the EGF‐Receptor (EGFR) by a blocking antibody and inhibition of the kinase domain of EGFR by a small molecule inhibitor (AG1478) led to a reduction in EGF induced directional sensing and migration velocity. Additionally, low, non‐effective single doses of antibody and small molecule inhibitor showed synergistically effective inhibition if combined. Those results support EGF being a potent guidance cue for MDA‐MB‐231 cell migration and help to understand the mechanisms behind chemotactic migration driven cancer metastasisThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SOCS3 down‐regulates ROS signaling and inhibits M1 and M2 human macrophage polarization
Authors: Lee Choong‐Eun , Yoon Hye‐young , Jeong Hana 
Published: June 2021

Initially found as feed‐back regulators of Jak/STAT pathways during cytokine signal transduction, the suppressors of cytokine signaling (SOCS) proteins have been recognized as modulators of inflammation through the regulation of macrophage activity. Macrophages are critical role players in immune‐inflammatory reactions to combat pathogen and to restore tissue homeostasis through the balanced action of distinct phenotypes: pro‐inflammatory M1 vs antiinflammatory M2. While anti‐inflammatory effect of SOCS1 through the inhibition TLR and NF‐kB signaling is well established, the role of SOCS3 in macrophage function has been controversial, being pro‐inflammatory or anti‐inflammatory. In the present work, we have studied regulatory functions of SOCS3 in inflammation employing M1 vs M2 differentiation systems and investigated the regulatory mechanisms involved. Human promonocytic THP1 cells were differentiated by PMA treatment and subject to polarization to M1 vs M2 phenotypes with LPS vs Dex stimulation, respectively for 24 to 48 h. Under such M1 or M2 differentiation condition, intracellular ROS generation and MAPK activation, albeit with different kinetics, were induced which preceded the pro‐inflammatory or anti‐inflammatory markers, respectively. Over‐expression of SOCS3 attenuated the LPS‐ or Dex‐induced ROS generation and suppressed both M1 and M2 differentiation as measured by the expression of proinflammatory (IL‐1beta, TNF‐alpha, and IL‐6) and anti‐inflammatory (TGF‐beta and IL‐10) cytokines. On the contrary, shRNA‐mediated SOCS3 knock‐down has shown the opposite effects. As a mechanism of ROS down‐regulation by SOCS3, we have observed the upregulation of anti‐oxidant factors such as Nrf‐2, Trx, SOD, and Prx in SOCS3 over‐expressing cells. Among MAPK members regulated by ROS signal, SOCS3 inhibited p38 activity critical for IL‐10 production during M2 differentiation with a prominent upregulation of MKP1. As SOCS3 suppressed STAT activation and GILZ expression important for M1 and M2 differentiation, the role of ROS signal in the regulation of these downstream transcription factors is also under investigation.Support or Funding Information< Supported by NRF grants 2015‐003291 and 2016‐911262 >This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The role of yeast RNA polymerase I initiation factor Core Factor subunit Rrn7 in promoter open complex formation
Authors: Jackobel Ashleigh J. , Han Yan , He Yuan , Knutson Bruce A. 
Published: June 2021

A central component in S. cerevisiae RNA polymerase I (Pol I) pre‐initiation complex (PIC) formation and transcriptional regulation is the evolutionarily conserved initiation factor termed Core Factor (CF). CF is a heterotrimer composed of Rrn6, Rrn11, and the TFIIB‐related factor Rrn7. Recent Cryo‐EM structures of the Pol I initiation complex reveal that CF resembles a right hand grasping promoter DNA in which key contacts are mediated by Rrn11 and Rrn7 (Han et al., eLife, 2017). Similar to its Pol II counterpart TFIIB, CF subunit Rrn7 consists of an N‐terminal zinc ribbon (ZR) domain followed by a linker region and two cyclin fold repeats. We systematically mutagenized the Rrn7 ZR and linker domains to further understand their role in Pol I transcription initiation. We found that ZR and linker mutants are still able to form the CF complex and assemble a PIC, yet are defective in promoter opening. These findings, paired with the Cryo‐EM structure, suggest that CF acts as a ratchet to drive promoter DNA into the active site of Pol I and that the Rrn7 ZR and linker domains play a dynamic role in ATP‐independent promoter opening.Support or Funding InformationThis work was supported by grants awarded to BAK: National Cancer Institute (5K22CA184235), Alexandrine and Alexander L. Sinsheimer Scholar award, and SUNY Research Foundation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The novel cardio‐oncology drug, NP‐6A4, regulates the growth of multiple breast cancer cell lines through distinct cell‐specific mechanisms
Authors: Pulakat Lakshmi , Belenchia Anthony M , Toedebusch Ryan G 
Published: June 2021

While chemotherapeutic agents (anthracyclins, kinase‐inhibitors) have improved 5‐year breast cancer survival, their cardiovascular toxicity cause alarming increase in life‐threatening heart disease for millions of cancer survivors. Therefore, developing new cardio‐oncology drugs to protect heart during and after chemotherapy is an immediate critical need. Although literature indicate that Angiotensin II type 2 receptor AT2R attenuates cardiac damage, to date there are no AT2R agonists that are used in clinic to treat heart disease. NP‐6A4 is an AT2R peptide agonist that has recently received orphan drug designation for pediatric cardiomyopathy from the FDA. Our previous studies indicated that NP‐6A4 treatment (1.8mg/kg/day) could improve cardiac function and attenuate cardiomyopathy in diabetic rats and protect human cardiovascular cells from acute nutrient stress. We hypothesized that since AT2R is a regulator of cell growth, NP‐ 6A4‐AT2R signaling will attenuate breast cancer cell growth. Here we report NP‐6A4 treatment of triple negative breast cancer cells (MDA‐MB‐231) and estrogen positive breast cancer cells (MCF‐7) resulted in significant (p<0.05) suppression of cell index (as determined by real time cell analyzer), and survival (as determined by MTS assay), albeit the underlying mechanisms were different. NP‐6A4 treatment of MCF‐7 cells augmented Tamoxifen‐induced inhibition of cell growth (p<0.05), and suppressed proliferation marker Ki67 (20%) and phosphokinases CREB (32%), Jun N‐terminal kinase (JNK) (51%) and Erk1/2 (41%). In MDA‐MB‐231 cells, NP‐6A4 treatment caused increase in apoptotic marker Caspase 3 (2.3‐fold increase), but suppressed its inhibitor phosphokinase p38 (37%) (p<0.05), and also suppressed phosphokinases CREB (30%), JNK (51%) and Erk1/2 (33%). However, doxorubicin (anthracycline)‐induced cell death was not reversed by NP‐6A4. NP‐6A4 treatment increased different tumor suppressors: ATIP 1 mRNA in MCF‐7 and ATIP‐3 mRNA in MDA‐MB‐231 (p<0.05). These data suggest that NP‐6A4 differentially regulates human breast cancer cell growth and has high potential as a synergistic cardio‐oncology drug for breast cancer treatment.Support or Funding InformationThis work was supported by NIH grants: 1R01HL118376‐01 and 1R01HL138988‐01A1This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cooperative Inhibition of Wnt/β‐Catenin Signaling by Klotho and Vitamin D: Implications for Chemoprevention
Authors: Khan Sameera , Khan Zainab , Whitfield Graham , Haussler Mark , Jurutka Peter 
Published: June 2021

The Klotho gene has been closely associated with delayed onset of aging. Inhibition of the klotho protein promotes aging‐like phenotypes, while its overexpression has been found to extend lifespan in mice. The single‐pass transmembrane isoform of klotho (m‐klotho) forms a receptor complex with the fibroblast growth factor (FGF) receptor to create a high‐affinity binding site for FGF23, a hormone involved in phosphate homeostasis and suppressing vitamin D activation. In contrast, secreted klotho (s‐klotho) forms via ectodomain shedding of the membrane‐bound isoform or through alternative splicing of the Klotho gene. It exhibits pleiotropic functions presumably via its weak glycosidase activity, including the inhibition of the insulin/insulin‐like growth factor 1. The proposed anti‐aging properties of s‐klotho have already been demonstrated by its endocrine role in reducing oxidative stress and promoting cardiovascular protection. Due to its potential role as a tumor‐suppressor, we have focused on how s‐klotho interacts with a known chemopreventative agent and steroid hormone, 1,25‐dihydroxyvitamin D3 (1,25 D), and its nuclear receptor (VDR) to inhibit the activity of target proto‐oncogenes. One such target for 1,25D/klotho interaction is β‐catenin. Mutations in the Wnt/β‐catenin signaling pathway have been implicated in numerous cancers, particularly colorectal neoplasia. In this study, the activity of β‐catenin was analyzed in vitro using transcriptional luciferase assays and western blotting as a response to modulated levels of exogenous klotho, 1,25D, and VDR. In addition, quantitative polymerase chain reaction (qPCR) was utilized to evaluate the ability of s‐klotho and 1,25D to inhibit the expression of known β‐catenin target genes including cell cycle regulators such as cyclin D1 and c‐myc, as well as the apoptosis inhibitor survivin. Our results revealed that s‐klotho stimulated VDR transactivation while inhibiting β‐catenin activity independently as well as synergistically with VDR and 1,25D. These findings were further supported by the 1,25D/klotho‐mediated down‐regulation of cyclin D1, c‐myc, and survivin expression. The dual isoforms of klotho exhibited functional differences, as lower concentrations of m‐klotho were found to boost β‐catenin activity, implicating the secreted isoform of klotho in directing its hypothesized anti‐aging properties. Our findings indicate that the crosstalk between vitamin D and klotho may promote healthful aging in part due to their cooperative tumor‐suppressive role and inhibition of factors implicated in oncogenic signaling.Support or Funding InformationThis project was funded by the New College of Interdisciplinary Arts and Sciences at Arizona State University and the National Institute of Health.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sestrin2 Supports Lung Tumor Growth in Early Stages of Cancer Development in Mice but Might Play a Tumor Suppressive Role in the Late Stages of Carcinogenesis
Authors: Budanov Andrei Vladimirovich , Ding Boxiao , Parmigiani Anita , Chumakov Peter Mikhailovich , Lee Jun Hee , Chawla Ayesha , Grossman Steven 
Published: June 2021

The goal of this study was to understand the role of the p53‐inducible SESN1 and SESN2 genes in the development of lung cancer. Our studies are based on the development of mouse models employing inducible activation of the Kras oncogene in mice deficient for Sesn1 or Sesn2 genes. These mice develop lung cancers in response to injection with Adeno‐Cre virus.The members of the mammalian Sestrin gene family (SESN1, SESN2 and SESN3) encode evolutionary conserved proteins called sestrins that play an important role in regulating cell viability and metabolism in response to stress. The major activity of Sestrins is modulation of mammalian target of rapamycin complexes 1 and 2 (mTORC1/2), which are responsible for regulation of cell growth, metabolism, autophagy and viability. Strikingly, two of three members of the mammalian sestrin family, i.e. SESN1 and SESN2, are targets of transcription factor p53 which plays a major role in tumor suppression. We found that both SESN1 and SESN2 genes are downregulated in the majority of human lung cancers. Therefore, we proposed that SESN1 and SESN2 might be responsible for tumor suppressive activities of p53. However, inactivation of a single Sesn2 gene or both Sesn1 and Sesn2 genes did not affect the number of tumors in mice. Moreover, we observed that Sestrin2‐positive tumors were larger, indicating that Sestrin2 supports tumor growth during early stages of lung carcinogenesis. These effects correlated with higher activity of AKT kinase in the Sestrin2‐proficient mice as compared to Sestrin2‐deficient counterparts. Being a major regulator of cell growth and proliferation, AKT can be a critical target of Sestrin2 in regulating of tumor growth. Nevertheless, we have also demonstrated that silencing of Sestrin2 in human lung adenocarcinoma cells accelerates growth of tumor xenografts in nude mice and provides resistance to anticancer genotoxic drugs.Therefore, we conclude that Sestrin2 support tumor growth at early stages of cancer development, but might suppress carcinogenesis in the late stages. Moreover, tumors with low expression of Sestrin2 may be selected during chemo‐ and radiotherapy.Support or Funding InformationThe studies were supported by the Grant 17‐14‐01420 from the Russian Science Foundation to AB and NCI CA172660 to SG.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modulatory effect of Cashew (Anacardium occidentale L.) nut supplemented diet on fertility activity of clomiphene citrate in male rats
Authors: Akomolafe Seun 
Published: June 2021

ContextIncreasing concern has been expressed about the declining sperm count of humans and the potential fertility effects of clomiphene citrate, a synthetic oestrogen‐antagonist on human reproductive health in the last few decades.AimThis study aims to investigate the influence of cashew nut supplemented diet on fertility activity of clomiphene citrate in male rats.MethodsThe rats were divided into six groups (n=6): rats fed basal diet; rats fed basal diet and then given clomiphene citrate (cc) orally; rats fed diet supplemented with 10% processed cashew nut and given cc orally; and rat fed diet supplemented with 20% processed cashew nut and given cc orally; rats fed diet supplemented with 10% processed cashew nut and rat fed diet supplemented with 20% processed cashew nut.ResultsThe results revealed that there was a significant (p<0.05) improvement of total testosterone level and epididymal sperm count, viability and progressive motility in all the groups, in comparison to control, with more significance in combination therapy group. Similarly, increased testicular and epididymal total antioxidant capacity (TAC), nitric oxide (NO), total thiol (T‐SH), nonprotein thiol (NPSH) levels, Glutathione‐S‐transferase (GST) and steroidogenic enzymes activities with concomitant decrease in reactive oxygen species (ROS) level, thiobarbituric acid reactive substances (TBARS) production and arginase activity were observed in all the groups when compared with control, with more significance in combination therapy group.ConclusionTherefore, the combination therapy with cashew nut boosts the fertility potentials of clomiphene citrate. Thus, this combination therapy can be used to treat male partners suffering from infertility.Support or Funding InformationnilThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CRY1 dysregulation leads to hyperglycemia in diabetic mice
Authors: KIM YEYOUNG , LEE GUNG , KIM JAEBUM 
Published: June 2021

Hepatic glucose production (HGP) is essential for maintenance of normal glucose homeostasis. Glucagon promotes hepatic gluconeogenesis to increase the level of blood glucose upon fasting. However, during the postprandial state, insulin suppresses HGP via FOXO1 regulation. In diabetes, HGP dysregulation and insulin resistance contributes to hyperglycemia. Recently, it has been reported that CRY1 could act as a potential suppressor of hepatic gluconeogenesis by FOXO1 protein degradation. We previously reported that the level of CRY1 protein was decreased in the liver of obese and diabetic animals. However, the underlying mechanism for hepatic CRY1 dysregulation has not been thoroughly elucidated. In order to find out which factors are involved in CRY1 degradation, we screened CRY1 binding proteins in the liver of diabetic mice. We discovered that several candidate proteins appear to physically associate with CRY1 to mediate its stability. Our data support that CRY1 binding proteins could regulate CRY1 protein stability, providing a clue to understand the mechanism for hyperglycemia through CRY1 dysregulation in the liver of diabetic mice.Support or Funding InformationThis work was supported by NRF of Korea Grant MSIP 2011‐0018312 and BK21.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Biomedical Status of Women Attending In‐Vitro Fertilization Clinics in Nigeria
Authors: Gbenle Oluwasegun Adekunle , Igwo‐Ezikpe Miriam N. , Taiwo Idowu A. , Anyashina Olumuyiwa A. , Ofoma Osita N. , Ikwuazom Okwunna O. 
Published: June 2021

Infertility constitutes a major problem in many developing countries like Nigeria where itsprevalence is significant. The relationship between the endocrinological, physiological and environmental determinants of conception in IVF among Nigerian infertile women have not been well elucidated; hence, the need for the biomedical status of women attending IVF Clinics in Nigeria.Blood samples of infertile women (n=200) undergoing in‐vitro fertilization were obtained from different fertility clinics in Nigeria. Questionnaires were distributed to participants to obtain physiological characteristics such as age, blood group, genotype and Rhesus factor. The levels of reproductive hormones (Luteinizing Hormone, Follicle Stimulating Hormone and Prolactin) were analysed using a micro‐plate reader while the levels of heavy metals (Mercury, Lead, Cadmium and Chromium) in the blood samples was determined using atomic absorption spectroscopy.There was significant (p< 0.05) difference in the levels of Luteinizing Hormone, Follicle Stimulating Hormone and Prolactin between the age groups of the infertile women. There was higher non significant (p>0.05) levels of the three reproductive hormones in the negative Rhesus factor subjects compared to positive Rhesus factor. However, there was no significant (p<0.5) difference in the three reproductive hormones in relation to the blood grouped subjects. There was significant (p<0.5) difference between the levels of the reproductive hormones and age. There were more negative correlations between the levels of reproductive hormones and levels of heavy metals whereas there was no significantdifference between the levels of heavy metals and age of infertile women.The study suggests that IVF patients with a specific blood group do not have a higher incidence of reduced fertility in response to ovarian stimulation. It also suggests no relationship between Rhesus factor and infertility but age and levels of heavy metals may affect female infertility.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of Alaskan Wild Bog Blueberries Anthocyanins in the Mitigation of Type II Diabetes.
Authors: Callahan Brandaise , Collin Aline , Kuhn Thomas 
Published: June 2021

Alaska natives were recently found to have the fastest increase in diagnosed type II diabetes (T2D) cases in the U.S. population due to the influence of western diets. A common manifestation of T2D is insulin resistance which alters the body's regulation of insulin and glucose metabolism. Alaskan blueberries have been shown to have a higher content of antioxidants in the form of anthocyanins compared to other varieties. Dietary anthocyanins have have shown to act on the insulin transduction pathway and reduce insulin resistance. Our research focuses on the anthocyanins present in Alaska wild bog blueberries and their potency to prevent and reduce insulin resistance and therefore mitigate T2D development.The experimental design encompassed differentiated 3T3‐L1 adipocytes exposed to palmitate for 16 hr, which reduces insulin‐dependent glucose uptake, in the presence or absence of anthocyanins. Our study focused on the phosphorylation of AS160, a pivotal substrate in the insulin transduction pathway, as a response of the cells to insulin. Anthocyanin fractions (ANC) (10 ug/mL) were applied for 1 hr prior to palmitate (0.75 mM, 16 h) and were present throughout the stress period. Total cell lysates were analyzed by western blotting to determine the presence of phosphorylated AS160.Each condition was run in duplicate and two western blots were run. The software ImageJ was used to obtain relative levels of p‐AS160. Values were obtained from the ratio of insulin stimulated over basal levels of AS160.The control, (no palmitate or ANCs), had the highest amount of phosphorylated AS160 (1.925, p <0.05). Cells stressed with palmitate had the least AS160 phosphorylation levels (0.791 p= 0.05). Importantly, cells treated with ANCs prior to stress exhibited 1.047 (p=0.11) of phosphorylated AS160.The data supports the hypothesis that the ANCs used prior to stressing the cells mitigates insulin resistance. Future work will look at an intervention paradigm and focus on a comparative analysis of prevention and intervention.Support or Funding InformationWork reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under three linked awards number RL5GM118990, TL4GM118992 and 1UL1GM118991. The work is solely the responsibility of the authors and does not necessarily represent the official view of the National Institutes of Health.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.