bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of Cyr61 signaling in solid tumor cells diminishes response to a histone deacetylase inhibitor: Challenges for HDACIs
Authors: Ghosh Arnab , Ghosh Priyanka , Maity Gargi , Banerjee Sushanta K , Banerjee Snigdha 
Published: November 2023

Background and ObjectiveHistone deacetylase (HDAC) inhibitors are clinically proven epigenetic‐based drugs for hematological cancers. However, the impact of these inhibitors on solid tumors is disappointing and debatable. Recent studies have shown that the solid tumor cell lines are less sensitive to a HDAC inhibitor Vorinostat (suberoylanilide hydroxamic acid; SAHA) as compared to hematological cell lines. However, it remains elusive why solid tumor cell lines responded weakly to HDAC inhibitors. Previously, it has been reported that CCN1/Cyr61 plays critical role in invasive front and drug resistance in breast and pancreatic cancer. Thus, our goal is to determine whether CCN1 signalling impair activity of SAHA in breast and pancreatic cancer cells.MethodsTo test this objective, various breast cancer cell lines (MCF‐7, ZR‐75‐1, MDA‐MB‐231, HCC‐70) and pancreatic cancer cell lines (BxPC‐3, AsPC‐1 and Panc‐1) were treated with SAHA with different does and times. Levels of CCN1 and its downstream signalling molecules were determined in SAHA‐treated or untreated cell extracts using Western blotting. In addition, cell viability, migration and sphere formation were examined in parental and CCN1‐depleted cells in the presence or absence of SAHA.Results and ConclusionsHere we have shown that CCN1 was upregulated in various breast and pancreatic cancer cell lines following SAHA treatment via epigenetic mechanism. Distribution of CCN1 in the nucleus and cytoplasm was also drastically altered in SAHA‐treated cells. Normally, CCN1 expression was detected in the cytoplasm. However, following SAHA treatment, CCN1 expression was located predominantly in the nucleus. Functionally, CCN1 had dual outcome in these cells. The cell viability assay indicated that CCN1‐positve breast cancer or pancreatic cancer cells were more sensitive to SAHA as compared to CCN1‐negative cells. CCN1 ablation by neutralizing antibody treatment in MDA‐MB‐231 cells significantly suppressed the inhibitory effect of SAHA in MDA‐MB231 cells. However, aggressive phenotype such as migration towards SDF‐1 and sphere‐formation we remarkedly elevated in SAHA treated cells as compared to untreated cells. These effects of SAHA were rescued by CCN1‐depletion. Collectively, these studies indicate that CCN1 activation limits the response to SAHA in solid tumor cells and thus suggesting that combination therapy of SAHA andCCN1‐inhibitor could be an ideal therapeutic approach to make HADAC inhibitor sensitive to solid tumors.Support or Funding InformationVA Merit Grant.
DDB2 Activates Rnf43 and Regulates Wnt/β‐catenin Signaling in Colorectal Cancer Cells
Authors: HUANG SHUO , Fantini Damiano , Merrill Bradley , Bagchi Srilata , Raychaudhuri Pradip 
Published: November 2023

The Wnt/β‐catenin signaling pathway plays important roles in the development and progression of colorectal cancer. However, its de‐regulation in colorectal cancer is not clearly understood because, in addition to activating pro‐cancer genes, this signaling pathway activates expression of several tumor suppressor genes, including Rnf43. Previously, we showed that DDB2 suppresses metastasis of colorectal cancer cells harboring activating mutations in the Wnt/β‐catenin signaling pathway. Here, we show that DDB2 is critical for activation of Rnf43, which regulates Wnt‐signaling by inhibiting the levels of Wnt‐receptors. DDB2 recruits β‐catenin onto an upstream regulatory region in the Rnf43 gene, and is required for its expression by the Wnt‐signaling pathway. Cells lacking sequences in the regulatory region of Rnf43 involved in DDB2‐binding or DDB2‐deficient cells are impaired in expressing Rnf43 and are insufficient to ubiquitinate the Wnt‐receptor FZD5; consequently, those cells exhibit greater response to Wnt3a ligands. The results identify DDB2 as a novel partner as well as a negative regulator of Wnt‐signaling. The results also provide insights into how loss of DDB2 drives metastasis of colorectal cancer.Support or Funding InformationNIH RO1‐CA175380
Modelling the Effects of Laser Photothermal Therapy on Proteins HSP70 and P53
Authors: Milcarek Andrew , Daus Kevin , Alpaugh Mary , Dobbins Tabbetha 
Published: November 2023

Gold nanoparticles (AuNPs) and nanorods (AuNRs) are able to conjoin with biological molecules and efficiently absorb light for conversion into heat energy. Thus, they are being investigated for treatment of a variety of cancers using lasers to induce the conversion to heat in a technique known as laser photothermal therapy (LPTT). Since its first demonstration in 2005, LPTT has been increasingly researched and its efficacy has been demonstrated in mouse models. However, details which focus on the effect of heat on surrounding healthy cells is extremely limited. Likewise, study of whether intracellular vs. extracellular AuNP and AuNR uptake is most beneficial (and for which cancer types) is not undertaken. In this research, we examine these two aspects of laser photothermal therapy with the research questions: (i) how does heat exposure regulate proteins within the surrounding cells, specifically HSP70 and P53 and (ii) how do the particles distribute themselves at the surface of and/or within the cell. Specific to objective (i), the proteins HSP70 and P53 are examined. HSP70 is a heat shock protein responsible for wound healing, particularly in thermal burns. P53 is the “tumor repair protein” which is responsible for regulating cell transcription. Assuring maintenance of normal concentrations and the correct conformation of these proteins in the surrounding healthy cells will be important for implementing LPTT in patient care. Specific to objective (ii), benign planar cell models, human embryonic kidney cells (HEK‐293) and primary mouse embryonic fibroblast cells (3T3), are studied with quantitative monitoring of particle uptake concentration and distribution on cell viability during the therapy treatment. Size, surface charge, and surface treatment determine whether particles will undergo cellular uptake. These and other protein additives (such as chitosan coatings) determine whether the particles will diffuse across the cell wall. Key to studying cellular uptake is the comparative examination of planar tumor HeLa cells and the 3D Mary‐X tumor cell model. Objective (ii) informs the LPTT community of researchers as to which AuNPs and AuNRs conditions enhance cellular uptake for tumor cells—without selective uptake in benign cells.Support or Funding InformationThe work is being carried out in Dr. Tabbetha Dobbins lab at Rowan University. Funding at this time provided by Rowan University Department of Biomedical and Translational Sciences.
Pim‐1 signaling in drug‐resistant colon cancer cells promotes cell survival and chemoresistance through up‐regulation of lactate production
Authors: Park Ga Bin , Kim Daejin 
Published: November 2023

Despite development of numerous target drugs, cancer drug resistance is still one of major obstacles to disturb anti‐cancer treatment in clinical field. Pim‐1 (Proviral integration site for Moloney murine leukemia virus‐1) kinase originally plays an essential role in cytokine‐induced signal transduction. However, overexpression of Pim‐1, as a proto‐oncogene, is also connected with cell proliferation, survival, and drug resistance. We found that the levels of Pim‐1 and mesenchymal markers in drug‐resistant colon cancer cells were highly expressed. To determine the effect of Pim‐1 on generating the chemoresistant cells, we investigated the influence of Pim‐1 on the expression of multi‐drug resistance gene. Targeted inhibition of Pim‐1 using small interference RNA (siRNA) significantly suppressed the up‐regulation of multiple drug resistance (MDR1), multidrug resistance protein 1 (MRP1), and cancer stem cell markers compared with the control group transfected with control siRNA. Silencing of Pim‐1 in chemoresistant HCT‐116 cells with siRNA resulted in downregulation of glycolysis‐associated enzymes, drug‐resistance related proteins, and a disintegrin and metalloproteinase (ADAM) family proteins. In addition, knockdown of Pim‐1 in OxR‐HCT‐116 cells triggered the ER stress‐mediated apoptosis through the loss of oxaliplatin or 5‐fluorouracil resistance. These finding suggest that Pim‐1‐associated signaling pathway plays a critical role in cancer progression and promoting chemoresistance of colon cancer cells.Support or Funding InformationThis study was supported by the Basic Science Research Program of Ministry of Education (NRF‐2015R1D1A1A01056672) and Ministry of Science, ICT & Future Planning (NRF‐2015R1C1A2A01053732) through the National Research Foundation (NRF) of Republic of Korea.
Genesis of Antibiotic Resistance XXIX: Clinical Perspectives of obdurate implementation of Antibiotics Time‐Out (ATO) in Mitigating Antibiotic Resistance Pandemic (ARP)
Authors: Anaya Edson , Martinez Juan , Calzoncit Melissa , Picazo Antonio , Blackmon Sarah , Jauregui Suzzette , Lechler Raul , Beautnagel Joanathan , Gutierrez Sonia , Johnson Dennie , Kannan Subburaj 
Published: November 2023

Outcomes of a service learning project, the Strategies for Successful Implementation of ATO, along with its impact in mitigating ARP is presented in this work. ATO strategies were followed as suggested by CDC, http://www.cdc.gov/getsmart/healthcare/implementation/checklist.html with modification in accordance with hospital regulations. ATO strategies were adopted before each surgical procedure to ensure every patient received the appropriate prescribed antibiotic before being taken to the operating room, and prior to the start of surgery. This strategy was employed and directed toward the prevention of nosocomial infections and used as a measure to ensure that congruency in patient and corrective procedure. Accordant with the review of success rates among patients who had completed the surgical modality, ATO strategies generated higher success rate (data not presented in accordance with/per HIPAA guidelines). Reevaluating the prescription of antibiotics and dosage(s) of antibacterial regimen in the context of specific geographical location has enabled the providers to rule out antibiotics that are not effective in a specific community. Another contributing factor to higher success rates in surgical outcomes is vigilant and strict enforcement of the dialogue between the surgical staff (Medicos), pharmacy administration, and the antibiogram (data on file – medical records) of each patient from clinical antibiotic sensitivity assay has enhanced the positive outcome of ATO. The combined review of surgical medicos, pharmacy, Center for Disease Control and Prevention alerts concerning the spread of AR pathogen, and information gleaned from the Sanford Guide to Antimicrobial Therapy has enabled the prescription of appropriate and accurate dosages to aid patients in recovery in a more prudent manner, before and after the surgical modality. On a comparative basis, this practice is similar to the grand rounds undertaken on periodic basis in the teaching hospitals across the globe. Taken together, effective implementation of the ATO as a routine practice in the surgical units (across specialties, e.g. Cardiothoracic Surgery, Transplantation Units) would potentially mitigate the ARP. Supplementary data analysis were from the Continuing Medical Education activities completed by Subburaj Kannan, M.D.(A.M) Ph.D. Data analysis will be presented at EB 2017.Support or Funding InformationProfessional development funds to Subburaj Kannan
Isolation, Expression, and Characterization of a β–lactamase produced by Meiothermus ruber
Authors: Grulke Luther , Bergstrom Alexander , Groen Tara , Analitis Elaina , Scott Lori , Crawford Patrick 
Published: November 2023

β‐lactamase enzymes are the most significant contributor to antibiotic resistance in bacteria. The pharmaceutical industry's response to antibiotic resistance has been to develop new antibiotics or β–lactamase inhibitors; these successes in the clinical setting require an understanding of the structure and function of the β–lactamases. A β–lactamase gene was identified in Meiothermus ruber, a thermophilic gram‐negative bacterium, with a bright red carotenoid pigment. M. ruber genomic DNA was utilized to clone the β–lactamase gene into the plasmid pKT1. The pKT1 plasmid contains a β–lactamase selection gene, which could potentially obfuscate kinetic assay data. Therefore, the β–lactamase gene from M. ruber was then amplified by PCR, inserted into pET‐26b, a protein‐expression vector containing a Kanamycin resistance selection gene, and transformed into E. coli. Restriction digestion and sequencing were performed on the successful transformants to verify the presence and correct orientation of the M.ruber β–lactamase gene. Activity and substrate preference of the gene product was determined by growing the bacteria in the presence of various β–lactam antibiotics. Preliminary tests suggest the M.ruber β–lactamase gene product can hydrolyze cephalosporin β–lactam antibiotics but not carbapenems. This result is supported by sequence comparison studies that suggest the gene product is a class C (AmpC‐type) β–lactamase. Further experiments are required to understand the structure and function of the β–lactamase from M.ruber.Support or Funding InformationThis work was funded by Augustana College Augustana College Summer Undergraduate Research Funds and the Department of Energy Joint Genome Institute's GEBA project.
Genesis of Antibiotic Resistance XXII: Mutagenic potential of the River Ganga (RG): Segment I, II, III‐a,b,c, vacate Hardy‐Weinberg‐Castle Equilibrium (HWCE), and concoct as a plausible contributor to an imminent Antibiotic Resistance Pandemic (ARP)
Authors: Francisco Martinez , Montoya Hector , Flores Jorge , Carbajal Carlos , Wickham Cheyenne , Lopez Christian , Johnson Dennie , Kannan Subburaj 
Published: November 2023

A comparative study of the water quality of City of Eagle Pass, Texas, Water Works and the River Ganga was conducted to determine the induction of antibiotic resistance (AR) in bacteria such as fecal coliform bacteria (FC) that are known to cause antibiotic resistance infectious diseases. The correlation and regression analysis was conducted to determine the dependency among the following variables; i. Nephelometric Turbidity Units (NTU) of RG Segment‐I, II, III a, III b, and III c, ii. Saprobic index (SI=S), iii. Incidence of Fecal Coliform (MPN/100ml), iv. Morbidity and Mortality rate due to AR pathogen induced infectious diseases. A linear, (positive) correlation coefficient was observed between increases in NTU (~40NTU) paralleled with higher FC count (F.C. (MPN/100ml): ~ 160000). Regression coefficient analysis (where y= a + bx) showed a slope=1. An average Biological Oxygen Demand (BOD) ~ above 15 (min 19 to max 76 TPD) from the inflow of untreated sewage drains ranging (min 7 to max 14) draining minimum 550 million liter/ day (MLD) to maximum of 1780 MLD along with Grossly Polluting Industries (GPI) maximum of 504 (segment IIIB) pose unknown quantities of mutagenic agents. A cumulative average S (S=∑ (s × h)/∑h) where s=saprobic index value of indicator species, h‐frequency of occurrence of each species values of RG I, II, III a, b, and c showed strong pollution load correlating with FC count and NTU. Regression coefficient analysis on three variables; NTU, S index, and FC count showed slope =1, where y=a+bx. A comparison of RG I, II, III a, III b, and III c versus RG IV a, and IV b indicated that both segments of RG showed a similar trend in pollution load. Analysis has shown that a similar FC count is determined in both sections as 719 GPI ‐ releasing pollutants (~3000MLD) in RG I, II, III a, III b, and III c compare to only 36 GPI ~3000MLD releasing ~2360 MLD in to RG IV a, and IV b, with the later having the highest FC count. Data on mortality and morbidity (M&M) due to incidence of AR pathogen induced infectious diseases in the regions spanning RG segment 1 through III, were incomplete and incoherent. This is attributed to either the indifference of state and federal agencies in the collection of M&M data or the absence of adequate resources. Taken together, compare to the CEPWW, water quality criteria the RG Segment I – III a, III b, and III c is patently polluted and pose an abysmal concern as an ever increasing and accumulating mutagenic potency with the propensity of inducing an increased mutation frequency in the AR gene pool of FC, thus contributing to an ARP. Data analysis to be presented in EB 2017.Support or Funding InformationProfessional development funds to Subburaj Kannan
Stimulation of astrocyte fatty acid oxidation increases spare respiratory capacity under nutrient‐deprived conditions
Authors: Sifuentes Mikaela , Lechleiter James 
Published: November 2023

Astrocytes offer increased resistance to toxic extracellular conditions and regulate the microenvironment surrounding neurons, presenting them as a key target for protective strategies against stroke and other metabolically‐intensive injuries. Our lab previously reported that thyroid hormone can increase astrocyte survival by stimulating fatty acid oxidation (FAO). We observe that under 24 hours of nutrient deprivation, the maximal oxygen consumption rate in astrocytes decreases significantly, but can be restored by thyroid hormone treatment in a dose‐dependent response after a 15 minute exposure. The rapid increase in spare respiratory capacity is blocked by inhibition of fatty acid transport into the mitochondria, indicating that prolonged glucose deprivation can be protected against using FAO as an alternative fuel source. Additionally, we find that starvation conditions induce the formation of large lipid droplets in astrocytes, possibly providing an endogenous source of fatty acids to be utilized during nutrient deprivation. Treatment of these astrocytes with thyroid hormone reduced lipid droplet size and concentration, in line with other reports that lipid droplets can be consumed through FAO. In conclusion, our observations support our hypothesis that thyroid hormone increases astrocyte viability by switching astrocyte mitochondrial function from a glucose‐dependent pathway to a FAO‐dependent one.Support or Funding InformationResearch supported by National Institutes of Health R01AG007218 and the American Heart Association Predoctoral Fellowship 16PRE27670002
Acute oxidative challenge induces glucose uptake through AMPK‐mediated NO production in skeletal muscle
Authors: Kellogg Dean L , McCammon Karen , Masters Bettie Sue , Roman Linda 
Published: November 2023

We investigated the cellular pathways underlying glucose uptake in skeletal muscle (SkM) in response to reactive oxygen species (ROS). These underlying mechanisms are critical to understanding exercise‐induced oxidative stress and metabolic disease. Both AMP‐activated kinase (AMPK) and Nitric Oxide (NO) mediate the adaptive responses to oxidative stress. The goal of this study is to delineate the specific relationships between AMPK signaling and NO production and the resultant effects on GLUT4 translocation during acute oxidative stress. Utilizing an in vitro model of SkM (C2C12 myotubes), we demonstrate that exogenous ROS stimulate AMPK activity, NO production and phosphorylation of SkM‐specific neuronal NO synthase (nNOSμ). Dorsomorphin (an AMPK inhibitor) blocks nNOS phosphorylation, suggesting that the increase in NO is AMPK‐dependent. Furthermore, ROS induce translocation of GLUT4 to the cell membrane, which is blocked by L‐NAME (a NOS inhibitor), indicating that this process is dependent on NO. Taken together, these findings reveal that AMPK‐mediated phosphorylation of nNOS plays a central role in mediating the response of SkM to oxidative stress.Support or Funding InformationULT1TR001120 from the National Center for Advancing Translation Sciences to LJR
S‐nitrosylation Is Responsible for Muscle Atrophy and Weakness in GNE Myopathy
Authors: Miyakawa Miho , Cho Anna , Malicdan May Christine V , Nishino Ichizo , Noguchi Satoru 
Published: November 2023

BackgroundSialic acids are the most abundant terminal monosaccharides on glycoconjugates of eukaryotic cell surfaces and are involved in a variety of cellular functions. UDP‐N‐acetylglucosamine‐2‐epimerase/N‐acetylmannosamine 6‐kinase is a rate‐limiting enzyme for sialic acid biosynthesis, which is encoded by GNE gene. Mutations of GNE causes GNE myopathy, which is debilitating autosomal recessive myopathy, and is clinically characterized by atrophy and weakness in distal skeletal muscles. Previously, we have evidenced that sialic acid deficiency plays a key role for the pathogenesis of this myopathy with the GNE myopathy model mice, presenting muscle atrophy and weakness followed by myofiber degeneration. The phenotype was attenuated by sialic acid supplementation. And also, we found the elevation of reactive oxygen species in skeletal muscles of the model mice and demonstrated a therapeutic effect of an anti‐oxidant, N‐acetylcysteine (NAC) on muscle atrophy as well.ObjectiveWe aimed to identify protein modification(s) produced by ROS, which cause muscle atrophy and weakness in GNE myopathy.MethodsFor the analysis on S‐nitrosylation status using frozen skeletal muscles, we switched S‐nitrosocysteines to biotin‐labeled cysteins and purified the S‐nirosylated proteins by Neutravidin beads, followed by LC‐tandem mass shotgun analyses or western blotting.ResultsWe found that protein S‐nitrosylation was increased in GNE myopathy muscles. By proteome analysis, more than 60 proteins were identified in the S‐nitrosylated group, which revealed that the major proteins were involved in muscle contraction (actin and myosins) and energy production (muscle creatin kinase, enolase 3 and aldolase). We also found that sialic acid supplement significantly decreased S‐nitrosylation of these proteins as much as NAC treatment did.DiscussionOur results suggest that hyposialylation leads to failure in ROS scavenging and promotes S‐nitroso modification to deteriorate muscle contraction. Therefore, ROS cancelling would be a promising strategy to prevent muscle weakness and atrophy as well as to cure GNE myopathy.Support or Funding InformationThis study was partially supported by Intramural Research Grant (28‐6) for Neurological and Psychiatric Disorders of NCNP, by Comprehensive Research on Disability Health and Welfare (H25‐Shinkei Kin‐Ippan‐004) from Japan Agency for Medical Research and Development, AMED and by JSPS KAKENHI (23390236).