bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD44+/CD25−/CD100+ Cells Contribute to the Development of Basal Epithelial‐like Metaplastic Breast Cancer in Adiponectin‐deficient MMTV‐PyVT Mice
Authors: Chen Jie , Cao Handi , Liu Jing , Tse Wai Choi , Lee In‐Kye , Vanhoutte Paul M. , Xu Aimin , Yu Wang 
Published: November 2023

Cancer development depends on the interactions between epithelial carcinoma cells and the stromal microenvironment. The present study investigated the role of adiponectin, an adipokine with potent tumor‐suppressive activities, in modulating the stromal microenvironment during breast cancer development in mice. Trans‐implantation of the stromal vascular fractions (SVF) of mammary adipose tissue from adiponectin‐deficient MMTV‐PyVT mice promotes the development of MDA‐MB‐231 human breast cancers in NOD/SCID mice, which exhibit the aggressive metaplastic basal epithelial‐like molecular features. In MMTV‐PyVT mammary tumors, adiponectin deficiency significantly decreases the density of CD3+ tumor‐infiltrating lymphocytes and enhances the SVF composition of CD44+/CD25−/CD100+ cells that are originated from thymus. Adiponectin plays an important role in T‐cell development and maturation partly by modulating the epithelial stromal microenvironment of thymus. Adiponectin deficiency prevents the positive selection of CD3+ T‐cells and the negative selection of CD44+CD25−CD100+ T‐cells by thymic nurse cells (TNCs). Thus, reduced production of CD3+ T‐cells as well as enhanced release of CD44+/CD25−/CD100+ cells from thymus into the circulation give rise to a tumor‐permissive microenvironment in mammary tissues of MMTV‐PyVT mice lacking adiponectin. Moreover, CD44+/CD25−/CD100+ cells represent a major metaplastic component contributing to the development of more aggressive basal epithelial‐like mammary tumors in MMTV‐PyVT or NOD/SCID mice implanted with human MDA‐MB‐231 breast cancer cells. Treatment with adiponectin restores thymic T‐cell maturation/selection process and prevents the development of metaplastic basal epithelial‐like breast cancer in MMTV‐PyVT mice. In summary, the present study reveals that adiponectin acts as an immunomodulatory stromal factor in both the thymus and mammary tumor microenvironment to elicit the multifaceted anti‐tumorigenic functions.Support or Funding InformationThis work was supported in part by grants from Seeding Funds for basic research of the University of Hong Kong, Research Grant Council Grants (17121714, HKU779712M, HKU780613M) of Hong Kong; And the National Basic Research Program of China (973 program 2015CB553603).
Calcium Sulfide (CaS) Nanostructure Treatment on Non‐Small Cell Lung Cancer
Authors: Forti Kevin Muñoz , Figueroa Maria , Torres Bryan , Bernard Faviola , Rivera Daniel , Castro Miguel E , Suarez Edu B 
Published: November 2023

Lung cancer is the number one cause of death among deaths due to cancer in the United States independent of gender. Current treatments focus on surgical removal of tumors and surround tissue, chemotherapy and radiation therapy. Side effects of the aforementioned treatments have been shown to be detrimental to a patient's health and wellbeing and are capable of causing secondary neoplasms. Recently, nanotechnology has shown promise in providing safer and more efficient therapy options for cancer patients. Our colleagues from the University of Puerto Rico at Mayagüez synthesized nanostructures composed of calcium sulfide (CaS). Due to both calcium and sulfur being incorporated in numerous biological pathways with the former being a mediator in apoptosis, we expect these nanostructures to be biocompatible and with low side effects. In this study, we evaluated the effect of CaS nanostructures on pulmonary adenocarcinoma cells by incubating ATCC CRL‐2124 cell line in the presence of CaS nanostructures [3.8 μM], Etoposide [10 μM], and DMSO (vehicle) for 24‐, 48‐, and 72‐hours after a single dose at time 0. We hypothesized that CaS nanostructures would suppress the cells’ proliferation by interfering with their cell cycle phases and increasing apoptosis. We performed cell‐based assays to study the cell cycle and apoptosis‐mediated cell death. There were no statistically significant observations after 24 hours in non‐cancerous cells (MRC5). Our results showed that at 48‐hours post treatment the progress of malignant cells from Sub‐G1 into G0/G1 was suppressed along with an increase in apoptosis (p≤0.05). After initial treatment with CaS nanoclusters we observed a built up of malignant cells in S phase at 24‐hours (p≤0.05). These results provide a platform for elucidating the mechanism of action of CaS interfering with proliferation, which may lead to new alternative therapies for lung cancer. Currently we are studying apoptosis mediators and calcium‐dependent pathways believed to be influenced by CaS in cancerous cells.Support or Funding InformationThis work was supported by: UPR‐PRISE Program NIH Grant #R25GM096955
Differential Effects of Diets Rich in Fats, Carbohydrates and/or Fructose on the Development of Hepatic Steatosis and Mitochondrial Dysfunction
Authors: GARCIA‐BERUMEN CLAUDIA I , ALEJANDRE‐BUITRON ALAN , MONTOYA‐PEREZ ROCIO , RODRIGUEZ‐OROZCO ALAIN R , CALDERON‐CORTES ELIZABETH , PIÑA‐ZENTELLA ROSA MARIA , RAMOS‐GOMEZ MINERVA , SAAVEDRA‐MOLINA ALFREDO , CORTES‐ROJO CHRISTIAN 
Published: November 2023

Non‐alcoholic fatty liver disease (NAFLD) comprises the progressive development of steatosis, steatohepatitis and cirrhosis. The progression of the disease is a multifactorial process involving risk factors such as obesity, diabetes mellitus, and dyslipidemia. Besides, NAFLD has been closely related with increased mitochondrial ROS production because excessive fat and carbohydrates dietary intake, with fructose playing a prominent role in this disease. The main goal of this work was to elucidate the contribution of these dietary components to the development of mitochondrial dysfunction and oxidative stress during NAFLD. We feed rats during 1 month with different diets containing standard diet, standard diet plus fructose (C+FR), high saturated fat plus high carbohydrates (HFHC), high saturated fat plus high carbohydrates plus fructose (HFHC+FR), only high fat (HF) and high fat plus fructose (HF+FR). A notable inhibition of mitochondrial respiration was observed with HFHC and HFHC+FR diets. This was associated to strong decline in the activity of complex I, particularly with the HFHC+FR diet. Rats feed with HFHC and HFHC+FR exhibited prominent ROS production. Moreover, the liver weight in the latter group increased significantly with respect to the other diets, indicating that this diet is the most harmful to liver health, which was further confirmed by an excessive fat accumulation seen in histopathological analyses. Therefore, it is concluded that the combination of saturated fat and carbohydrates in excess favors the development of mitochondrial dysfunction and ROS production, with the addition of fructose to this diet accelerating the early development of liver damage.Support or Funding InformationSupported by grants from Programa de Investigación 2016 de la Coordinación de la Investigación Científica de la UMSNH (CIC‐UMSNH) (CCR).
EXCITATION‐METABOLISM COUPLING: ROLE OF MEMBRANE DEPOLARIZATION ON THE MITOCHONDRIAL FUNCTION
Authors: Díaz‐Vegas Alexis , Cordova Alex , Utreras Yildy , Rosales Giovanni , Llanos Paola , Hidalgo Cecilia , Arias Manuel , Jaimovich Enrique , Contreras‐Ferrat Ariel Eduardo 
Published: November 2023

IntroductionDepolarizing stimuli induce signaling pathways in muscle that have pleiotropic effects. Two new voltage‐dependent phenomena, excitation‐transcription and excitation‐metabolism coupling (EMC), add to the canonical excitation‐contraction coupling process. The mechanisms involved in EMC, however, remain poorly understood.Material and MethodsMice were used at 6–8 weeks. Changes in mitochondrial and cytoplasmic Ca2+ levels were evaluated in living fibers from Flexor digitorum brevis muscle via imaging of fibers transfected with suitable plasmids. The mitochondrial potential and oxygen consumption were evaluated with Seahorse system and TMRE+ respectively.ResultsDepolarization of skeletal muscle fibers increased the cytoplasmic and mitochondrial Ca2+ levels, as detected using R‐CaM and CEPIA‐mt3, respectively. The depolarization‐dependent mitochondrial Ca2+ increase was partly prevented by dantrolene (50 μM), xestospongin B (10 μM) or IP3R1 knockdown, and was completely suppressed when using both inhibitors. Fibers pre‐incubated with apyrase (2 U/mL) displayed partially inhibited stimulation‐induced mitochondrial Ca2+ increase. Muscle cells were pre‐incubated with caged‐IP3 (5 μM); photo‐release of IP3 induced a transient increase in mitochondria Ca2+ levels that was inhibited by xestospongin B infibers. Finally, depolarization incrased mitocrondrial membrane potencial and oxygen consumption and these effects were partially reduced by IP3R knockdown.DiscussionAll these results suggest that activation of IP3R1 and RyR1 mediated Ca2+ release contribute to the mitochondrial Ca2+ increase and mitochondrial function produced by muscle activation.Support or Funding InformationFONDECYT (11130267, 1151293), PIA‐ACT1111, CONICYT 21150604
Determining Kinetics of Reversible and Irreversible Biofilm Formation by Staphylococcus aureus and Pseudomonas aeruginosa
Authors: Tran Laura , Heuertz Rita 
Published: November 2023

Biofilms have become increasingly problematic in efficacious treatment of patient infections due to their tenacious survival, association with chronic and medical device‐associated infections, and resistance to antimicrobial treatments. According to the National Institutes of Health, biofilms play a role in approximately 80% of all chronic infections. This study focused on assessment of biofilm formation by bacterial models with P. aeruginosa, being the Gram negative model, and S. aureus as the Gram positive model. Inherent with the Gram positive model selected was the ability to discern biofilm characteristics of methicillin‐resistant S. aureus (MRSA) and methicillin‐susceptible strains (MSSA). The research design was use of a crystal violet‐based quantitative biofilm microplate assay performed at different time points (2–24 hours) on different strains of the above‐mentioned bacteria. To discern reversible attachment from irreversible adherence, wash conditions at two points in the procedure (one after bacteria were incubated and microwells were coated with bacterially‐produced biofilm; and the other after crystal violet staining of the biofilm) were changed as to number and rigor. Fewer number and less rigorous wash steps allowed for reversible attachment whereas greater number and more rigorous washes allowed for firmly adherent biofilm to be quantitated. Results indicated that MRSA produced higher amounts of biofilm than MSSA at most time points but both followed similar kinetic patterns (e.g., peaks at 8 h). Assessing MRSA and MSSA together, S. aureus showed peak biofilm at 6–8 hours and P. aeruginosa produced a similar magnitude of biofilm as S. aureus. Regarding number and rigor of wash steps, overall magnitude of biofilm was much lower when greater number and more rigorous washes were employed rather than fewer, less rigorous washes. These results indicate that number and rigor of wash steps was critical in quantitation of irreversibly attached biofilm.Support or Funding InformationFinancial support for LAT was received from the DeNardo Education and Research Foundation.
Ascorbate anti‐cancer activity is increased by Mn(III)N‐alkylpyridylporphyrins through production of reactive oxygen species
Authors: Bader Bader Hassan , Tovmayasan Artak , Craik James , Benov Ludmil , Batinic Haberle Ines 
Published: November 2023

Ascorbate (Asc) has shown potential for cancer therapy, and cancer cells typically accumulate porphyrins and display aberrant redox homeostasis. Mn(III)N‐alkylpyridylporphyrins (MnPs) are redox‐active superoxide dismutase mimetics with many prospective therapeutic applications. This study examined how structural modifications of MnPs might affect their anticancer activity through selective cellular targeting of production of reactive oxygen species (ROS) produced by redox‐cycling with ascorbate.MethodsPII/MDA ER‐negative human breast cancer and non‐tumorigenic HBL100 human breast epithelial cell lines were used as model systems. Uptake of MnPs and ascorbate oxidation rates were determined spectrophotometrically. MTT/SRB assays were used to investigate cytotoxic and anti‐proliferative actions of MnPs and Asc, and combinations thereof. Membrane damage and cell death mechanisms were investigated by flow cytometry.ResultsAscorbate and MnPs redox‐cycle to generate cytotoxic ROS, mainly H2O2. Anticancer efficiency of MnPs correlated with ability to oxidize ascorbate; differences in cellular uptake and subcellular distribution had only minor effect.ConclusionsThe major factor determining MnPs’ anticancer activity in cell culture models is redox potentialSupport or Funding InformationSupport: College of Graduate Studies, Kuwait University and Research Sector, Kuwait University; Research Grant YM04/14 and University Grant SRUL02/13
Effects of in vivo knockdown of a Cold‐Inducible RNA Binding Protein (CIRP) on Breast Tumorigenesis
Authors: Ochoa Joey L , Lujan Daniel A , Peterson Melanie N , Hathaway Helen J , Hartley Rebecca S 
Published: November 2023

RNA binding proteins (RBPs) are essential to processes that involve RNAs, beginning from their transcription through to their degradation. In this study, we utilize the polyomavirus middle T antigen (PyMT) mouse model to evaluate the role of cold‐inducible RBP (CIRP) in breast tumorigenesis. When PyMT is expressed in the mammary epithelium of mice, it causes aggressive tumorigenesis. CIRP has been shown to incite the translation of mRNAs encoding stress‐induced proteins, and is observed to have increased expression in breast cancer. We have shown that human CIRP overexpression in the PyMT mouse appears to inhibit breast tumorigenesis during early and late stages. Based on these results, the current study set out to evaluate the effects of CIRP knockdown on tumor development. We hypothesize that CIRP knockdown will allow greater progression of tumor growth, as well as a higher rate of metastasis. To address this hypothesis, CIRP was knocked down using siRNA in a PyMT ‐mammary tumor‐derived cell line. PyMT cells transiently transfected with siCIRP and siControl were injected into the mammary fat pad of a wild type mouse, and tumor growth assessed. CIRP knockdown in PyMT cells is being analyzed via RT‐qPCR and western blotting to determine the effective timing tor mammary fat pad injections. Tumor growth kinetics, final tumor burden and metastasis will be determined, as will expression of CIRP targets, proliferation, apoptosis and histopathological analysis. The results of this study will tell us if CIRP overexpression has the potential to protect against breast cancer.Support or Funding Information

Initiatives to Maximize Student Development (IMSD GM‐0602010)
Grant from the UNM‐HSC allocation Committee (RAC)
Structural Impact of Phosphorylation on the Dynamic Properties of SIKE
Authors: Minzer Ian , Machek Megan , Standen Heather , Kim Seung‐Hwan , Latzer Joachim , Bell Ellis , Bell Jessica 
Published: November 2023

Suppressor of IKKe (SIKE)inhibits TBK1 mediated phosphorylation of interferon regulatory factor 3 (IRF3) and misregulation of TBK1 is involved in both autoimmune disorders and cancer. Our work (Marion et al, J.Biol.Chem 288, 18612–23, 2013) demonstrated that TBK1 can phosphorylate SIKE in up to 6 positions, altering its ability to inhibit TBK1. Little however is known concerning the three dimensional structure of SIKE. Clustal analysis of available SIKE sequences suggests that some phosphorylation sites are highly conserved, and in comparison with WIT3 may be involved more specifically with SIKE function. Molecular dynamics studies using models created with Phyre2 are exploring the impact of differently phosphorylated forms of SIKE on the structure and dynamics of the molecule. C‐terminal Histidine tagged SIKE has been expressed and purified using Nickel‐NTA chromatography and limited proteolysis in conjunction with tandem mass spectrometry is being used to explore both the validity of the models and potential differences in dynamic structure of the molecule induced by phosphorylation.Support or Funding InformationThis work was supported by NIH grant R21A1 107447 to JKB
Conformational Stability of SIKE and a Mimic of a Maximally Phosphorylated Form
Authors: Graham Sasha , Kim Seung‐Hwan , Bell Ellis , Bell Jessica 
Published: November 2023

Suppressor of IKKe (SIKE)inhibits TBK1 mediated phosphorylation of interferon regulatory factor 3 (IRF3) and misregulation of TBK1 is involved in both autoimmune disorders and cancer. Our work (Marion et al, J.Biol.Chem 288, 18612–23, 2013) demonstrated that TBK1 can phosphorylate SIKE in up to 6 positions, altering its ability to inhibit TBK1. Little however is known concerning the three dimensional structure or stability of SIKE or the effects of phosphorylation. SIKE (his tagged) has been expressed and purified using Nickel NTA affinity chromatography and shown to be homogenous by MALDI‐tof mass spectrometry. To determine the effects of phosphorylation on the conformational stability of the protein, SIKE and SIKE‐S6E (where the 6 phosphorylatable serines have been mutated to the phosphomimic, glutamate) have been used in Fluorescence based Thermal Shift assays and chemical denaturation studies using a comparison of Urea and Guanidine Hydrochloride denaturation to examine the relative contributions of charge interactions in the overall stability of the two molecules. Results of direct experiment are being compared with Molecular Dynamics simulations using homology models created using Phyre2 for SIKE, SIKE S6E and a form with six phosphoserines built into the model.Support or Funding InformationThis work was supported by NIH grant R21A1 107447 to JKB
UDCA Prevents 17α‐Ethinylestradiol‐induced Cholestatic Injury via Inhibiting AMP‐activated Protein Kinase Activation
Authors: Li Xiaojiaoyang , Liu Runping , Yuan Zihang , Jiang Zhenzhou , Zhang Luyong , Zhou Huiping 
Published: November 2023

IntroductionCholestasis, as a common liver injury in clinic, increases the risk of other malignant hepatobiliary diseases such as hepatic carcinoma and cholangiocarcinoma. Estrogen‐induced cholestasis occurred in many susceptible women due to pregnancy or hormone therapy. Previous studies from our group and others suggest that AMP‐activated protein kinase (AMPK) induced phosphorylation of farnesoid X receptor (FXR), a key nuclear receptor involved in regulating bile acid homeostasis represents an important underlying mechanism of estrogen‐induced cholestasis. Most recent clinical study indicated that ursodeoxycholic acid (UDCA) monotherapy is effective in the treatment of intrahepatic cholestasis of pregnancy. However, the functional molecular/cellular mechanisms of UDCA‐mediated protective effect against estrogen‐induced cholestasis remains unclear and is the focus of this study.MethodsThe sandwich‐cultured rat primary hepatocytes and 17α‐ethinylestradiol (EE)‐induced rat cholestasis models were used. The protein levels were determined by Western blot analysis. The mRNA levels of targeted genes were determined by real‐time RT‐PCR. Immunofluorescent staining was used to detect canalicular structure of liver.ResultsEE significantly suppressed the expression of FXR and bile acid transporters via activation of AMPK signaling pathways. In rat primary hepatocytes, UDCA significantly inhibited EE‐induced AMPK activation and down‐regulation of FXR and bile acid transporters. UDCA rapidly and dose‐dependently induced phosphorylation of protein kinase A (PKA), which was correlated to its effect on inhibition of EE‐induced AMPK activation and down‐regulation of FXR expression. H89, a PKA inhibitor, blocked UDCA‐induced inhibitory effect on EE‐induced AMPK activation and downregulation of FXR and bile acid transporters. Furthermore, UDCA inhibited EE‐induced AMPK activation and downregulation of FXR and bile acids transporters in vivo rat cholestasis models.ConclusionUDCA protected hepatocytes from EE‐induced cholestatic injury via inhibiting AMPK activation. Our data also suggested that activation of PKA may also contribute to UDCA‐mediated protective effects.Support or Funding InformationThis poster presentation was supported by the Fundamental Research Funds for the Central Universities. And this study was supported by National Natural Science Foundation of China Grants (81320108029).
Quantification by ELISA of the Levels of DNA Topoisomerases I, IIα, and IIβ in the Nuclei of HuT 78 Cells Treated with Interleukin‐2
Authors: Foglesong Paul David , Nair R. Sajana , Gupta Nikita K. 
Published: November 2023

The activities of DNA topoisomerase I, DNA topoisomerase IIα, and DNA topoisomerase IIβ are stimulated in three transient peaks in nuclear extracts of HuT 78 cells, a human T cell lymphoma cell line, within 12 hrs after treatment of the cells with 1,000 units of interleukin‐2 (IL‐2)/ml. This study examined if the changes in the specific activities of these three enzymes was due to variations in the amounts of the proteins. The amounts of each of these enzymes in the nuclear extracts were determined by quantitative ELISAs using specific monoclonal antibodies, and the results were compared to the specific activities of the enzymes. There is no direct correspondence in the amounts of DNA topoisomerase I or DNA topoisomerase IIβ with their specific activities in the 12 hours following IL‐2 treatment of the cells. Therefore, increased synthesis/degradation of these proteins cannot explain the observed variations in their specific activities in the nuclear extracts of HuT 78 cells treated with IL‐2. However, the amount of DNA topoisomerase IIα increased after treatment with IL‐2 with a peak observed at 6 hours after IL‐2, but the amounts of the enzyme did not correlate with its specific activities. These results indicate that the mechanism for the observed activation of DNA topoisomerase I and DNA topoisomerase IIβ is not synthesis/degradation of the proteins, but rather post‐translational modification. However, the changes in the activities of DNA topoisomerase IIα in the extracts appear to be due to both increased synthesis/degradation of that enzyme and also post‐translational modification. It is hypothesized that phosphorylation of these proteins by protein kinases activated by IL‐2 is the primary mechanism for the observed variations in their specific activities.Support or Funding InformationThis work was supported by grants from the William & Ella Owens Medical Research Foundation and University of the Incarnate Word.
Glutathione S‐Transferase Expression in Mice (Mus musculus) Exposed in utero to Atrazine
Authors: Hafner Megan , Martin Debra 
Published: November 2023

Atrazine is a widely used herbicide on crops in the USA and has been found in ground water and streams where it does not easily breakdown. Although the EPA has set the annual safe concentration of atrazine in water systems to be 3 ppb, levels above this have been measured leading to concerns about environmental affects on health. In mammals, the liver is the organ that helps to breakdown environmental toxins, such as atrazine. One of the proteins of interest in the liver is glutathione S‐transferase (GST) because of its ability to target xenobiotics. In this study, GST levels were determined by Immunoblots for liver and serum samples of new born mice (Mus musculus) pups who were exposed to 0, 3, and 30 ppb atrazine in utero (n= >20 per group). Results demonstrated that there was a decreasing trend in liver GST levels as exposure to Atrazine in utero increased suggesting potential liver damage. This was not supported by the serum samples since they also showed a significant decrease in GST level as exposure to Atrazine in utero increased.Support or Funding InformationR&D SystemsSaint Mary's University of Minnesota