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      1. Author :
        Al‐Dujaili, Saja A; Koh, Amy J; Dang, Ming; Mi, Xue; Chang, Wenhan; Ma, Peter X; McCauley, Laurie K
      2. Title :
        Calcium Sensing Receptor Function Supports Osteoblast Survival and Acts as a Co‐Factor in PTH Anabolic Actions in Bone
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Journal of cellular biochemistry
      6. Products :
      7. Volume :
        117
      8. Issue :
        7
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS; IVIS BLI in vivo
      12. Abstract :
        Anabolic actions of PTH in bone involve increased deposition of mineralizing matrix. Regulatory feedback of the process may be important to maintain calcium homeostasis and, in turn, calcium may inform the process. This investigation clarified the role of calcium availability and the calcium sensing receptor (CaSR) in the anabolic actions of PTH. CaSR function promoted osteoblastic cell numbers, with lower cell numbers in post-confluent cultures of primary calvarial cells from Col1-CaSR knock-out (KO) mice, and for calvarial cells from wild-type (WT) mice treated with a calcilytic. Increased apoptosis of calvarial cells with calcilytic treatment suggested CaSR is critical for protection against stage-dependent cell death. Whole and cortical, but not trabecular, bone parameters were significantly lower in Col1-CaSR KO mice versus WT littermates. Intact Col1-CaSR KO mice had lower serum P1NP levels relative to WT. PTH treatment displayed anabolic actions in WT and, to a lesser degree, KO mice, and rescued the lower P1NP levels in KO mice. Furthermore, PTH effects on whole tibiae were inhibited by osteoblast-specific CaSR ablation. Vertebral body implants (vossicles) from untreated Col1-CaSR KO and WT mice had similar bone volumes after 4 weeks of implantation in athymic mice. These findings suggest that trabecular bone formation can occur independently of the CaSR, and that the CaSR plays a collaborative role in the PTH anabolic effects on bone
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10413
      15. Serial :
        21048
      1. Author :
        Alfonso-Loeches, Silvia; Ureña-Peralta, Juan; Morillo-Bargues, Mª José; Gómez-Pinedo, Ulises; Guerri, Consuelo
      2. Title :
        Ethanol-Induced TLR4/NLRP3 Neuroinflammatory Response in Microglial Cells Promotes Leukocyte Infiltration Across the BBB
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Neurochemical research
      6. Products :
      7. Volume :
        41
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        TLR4; Alcohol; Microglia; Leukocyte; infiltration; Matrix metalloproteinase; BBB; IVIS; IVIS FLI
      12. Abstract :
        We reported that the ethanol-induced innate immune response by activating TLR4 signaling triggers gliosis and neuroinflammation. Ethanol also activates other immune receptors, such as NOD-like-receptors, and specifically NLRP3-inflammasome in astroglial cells, to stimulate caspase-1 cleavage and IL-1b and IL-18 cytokines production. Yet, whether microglia NLRs are also sensitive to the ethanol effects that contribute to neuroinflammation is uncertain. Using cerebral cortexes of the chronic alcohol-fed WT and TLR4-/- mice, we demonstrated that chronic ethanol treatment enhanced TLR4 mediated-NLRP3/Caspase-1 complex activation, and upregulated pro-inflammatory cytokines and chemokines levels. Ethanol-induced NLRP3-inflammasome activation and mitochondria-ROS generation were also observed in cultured microglial cells. The up-regulation of CD45high/ CD11b? cell populations and matrix metalloproteinase-9 levels was also noted in the cortexes of the ethanol-treated WT mice. Notably, elimination of the TLR4 function abolished most ethanol-induced neuroinflammatory effects. Thus, our results demonstrate that ethanol triggers TLR4- mediated NLRP3-inflammasome activation in glial cells, and suggest that microglia stimulation may compromise the permeability of blood–brain barrier events to contribute to ethanol-induced neuroinflammation and brain damage.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10379
      15. Serial :
        21047
      1. Author :
        Alsulaiman, Mona; Bais, ManishV; Trackman, PhilipC
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Journal of Cell Communication and Signaling
      6. Products :
      7. Volume :
        10
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Lysyl oxidase; Lysyl oxidase propeptide; Osteoblast; Osteoclast; Differentiation; Bone; IVIS; IVIS BLI in vivo
      12. Abstract :
        Lysyl oxidase pro-enzyme is secreted by tumor cells and normal cells as a 50 kDa pro-enzyme into the extracellular environment where it is cleaved into the ~30 kDa mature enzyme (LOX) and 18 kDa pro-peptide (LOX-PP). Extracellular LOX enzyme activity is required for normal collagen and elastin extracellular cross-linking and maturation of the extracellular matrix. Extracellular LOX-PP acts as a tumor suppressor and can re-enter cells from the extracellular environment to induce its effects. The underlying hypothesis is that LOX-PP has the potential to promote bone cell differentiation, while inhibiting cancer cell effects in bone. Here we investigate the effect of LOX-PP on bone marrow cell proliferation and differentiation towards osteoblasts or osteoclasts, and LOX-PP modulation of prostate cancer cell conditioned media-induced alterations of proliferation and differentiation of bone marrow cells in vitro. Effects of overexpression of rLOX-PP in DU145 and PC3 prostate cancer cell lines on bone structure in vivo after intramedullary injections were determined. Data show that prostate cancer cell conditioned media inhibited osteoblast differentiation in bone marrow-derived cells, which was reversed by rLOX-PP treatment. Prostate cancer conditioned media stimulated osteoclast differentiation which was further enhanced by rLOX-PP treatment. rLOX-PP stimulated osteoclast differentiation by inhibiting OPG expression, up-regulating CCN2 expression, and increasing osteoclast fusion. In vivo studies indicate that rLOX-PP expression by PC3 cells implanted into the tibia of mice further enhanced PC3 cell ability to resorb bone, while rLOX-PP expression in DU145 cells resulted in non-significant increases in net bone formation. rLOX-PP enhances both osteoclast and osteoblast differentiation. rLOX-PP may serve to enhance coupling interactions between osteoclasts and osteoblasts helping to maintain a normal bone turnover in health, while contributing to bone abnormalities in disease.
      13. URL :
        http://dx.doi.org/10.1007/s12079-015-0311-9
      14. Call Number :
        PKI @ user @ 10472
      15. Serial :
        21046
      1. Author :
        Alvarez, Silvia; Diaz, Marcos; Flach, Johanna; Rodriguez-Acebes, Sara; Lopez-Contreras, Andres J.; Martinez, Dolores; Canamero, Marta; Fernandez-Capetillo, Oscar; Isern, Joan; Passegue, Emmanuelle; Mendez, Juan
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Nat Commun
      6. Products :
      7. Volume :
        6
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS
      12. Abstract :
        Replicative stress during embryonic development influences ageing and predisposition to disease in adults. A protective mechanism against replicative stress is provided by the licensing of thousands of origins in G1 that are not necessarily activated in the subsequent S-phase. These /`dormant/' origins provide a backup in the presence of stalled forks and may confer flexibility to the replication program in specific cell types during differentiation, a role that has remained unexplored. Here we show, using a mouse strain with hypomorphic expression of the origin licensing factor mini-chromosome maintenance (MCM)3 that limiting origin licensing in vivo affects the functionality of hematopoietic stem cells and the differentiation of rapidly-dividing erythrocyte precursors. Mcm3-deficient erythroblasts display aberrant DNA replication patterns and fail to complete maturation, causing lethal anemia. Our results indicate that hematopoietic progenitors are particularly sensitive to replication stress, and full origin licensing ensures their correct differentiation and functionality.
      13. URL :
        http://dx.doi.org/10.1038/ncomms9548
      14. Call Number :
        PKI @ user @ 10238
      15. Serial :
        21045
      1. Author :
        Ames, Erik; Canter, Robert J; Grossenbacher, Steven K; Mac, Stephanie; Chen, Mingyi; Smith, Rachel C; Hagino, Takeshi; Perez-Cunningham, Jessica; Sckisel, Gail D; Urayama, Shiro
      2. Title :
        NK Cells Preferentially Target Tumor Cells with a Cancer Stem Cell Phenotype
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        The Journal of Immunology
      6. Products :
      7. Volume :
        195
      8. Issue :
        8
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS; IVIS BLI in vivo
      12. Abstract :
        Increasing evidence supports the hypothesis that cancer stem cells (CSCs) are resistant to antiproliferative therapies, able to repopulate tumor bulk, and seed metastasis. NK cells are able to target stem cells as shown by their ability to reject allogeneic hematopoietic stem cells but not solid tissue grafts. Using multiple preclinical models, including NK coculture (autologous and allogeneic) with multiple human cancer cell lines and dissociated primary cancer specimens and NK transfer in NSG mice harboring orthotopic pancreatic cancer xenografts, we assessed CSC viability, CSC frequency, expression of death receptor ligands, and tumor burden. We demonstrate that activated NK cells are capable of preferentially killing CSCs identified by multiple CSC markers (CD24+/CD44+, CD133+, and aldehyde dehydrogenasebright) from a wide variety of human cancer cell lines in vitro and dissociated primary cancer specimens ex vivo. We observed comparable effector function of allogeneic and autologous NK cells. We also observed preferential upregulation of NK activation ligands MICA/B, Fas, and DR5 on CSCs. Blocking studies further implicated an NKG2D-dependent mechanism for NK killing of CSCs. Treatment of orthotopic human pancreatic cancer tumor-bearing NSG mice with activated NK cells led to significant reductions in both intratumoral CSCs and tumor burden. Taken together, these data from multiple preclinical models, including a strong reliance on primary human cancer specimens, provide compelling preclinical evidence that activated NK cells preferentially target cancer cells with a CSC phenotype, highlighting the translational potential of NK immunotherapy as part of a combined modality approach for refractory solid malignancies.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10100
      15. Serial :
        21044
      1. Author :
        Andersen, Rikke K; Zaher, Walid; Larsen, Kenneth H; Ditzel, Nicholas; Drews, Katharina; Wruck, Wasco; Adjaye, James; Abdallah, Basem M; Kassem, Moustapha
      2. Title :
        Association between in vivo bone formation and ex vivo migratory capacity of human bone marrow stromal cells
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Stem Cell Research & Therapy
      6. Products :
      7. Volume :
        6
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS; IVIS BLI in vivo
      12. Abstract :
        Introduction: There is a clinical need for developing systemic transplantation protocols for use of human skeletal stem cells (also known bone marrow stromal stem cells) (hBMSC) in tissue regeneration. In systemic transplantation studies, only a limited number of hBMSC home to injured tissues suggesting that only a subpopulation of hBMSC possesses “homing” capacity. Thus, we tested the hypothesis that a subpopulation of hBMSC defined by ability to form heterotopic bone in vivo, is capable of homing to injured bone. Methods: We tested ex vivo and in vivo homing capacity of a number of clonal cell populations derived from telomerized hBMSC (hBMSC-TERT) with variable ability to form heterotopic bone when implanted subcutaneously in immune deficient mice. In vitro transwell migration assay was used and the in vivo homing ability of transplanted hBMSC to bone fractures in mice was visualized by bioluminescence imaging (BLI). In order to identify the molecular phenotype associated with enhanced migration, we carried out comparative DNA microarray analysis of gene expression of hBMSC-derived high bone forming (HBF) clones versus low bone forming (LBF) clones. Results: HBF clones were exhibited higher ex vivo transwell migration and following intravenous injection, better in vivo homing ability to bone fracture when compared to LBF clones. Comparative microarray analysis of HBF versus LBF clones identified enrichment of gene categories of chemo-attraction, adhesion and migration associated genes. Among these, platelet-derived growth factor receptor (PDGFR) α and β were highly expressed in HBF clones. Follow up studies showed that the chemoattractant effects of PDGF in vitro was more enhanced in HBF compared to LBF clones and this effect was reduced in presence of a PDGFRβ-specific inhibitor: SU-16 f. Also, PDGF exerted greater chemoattractant effect on PDGFRβ+ cells sorted from LBF clones compared to PDGFRβ- cells. Conclusion: Our data demonstrate phenotypic and molecular association between in vivo bone forming ability and migratory capacity of hBMSC. PDGFRβ can be used as a potential marker for the prospective selection of hBMSC populations with high migration and bone formation capacities suitable for clinical trials for enhancing bone regeneration.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 10214
      15. Serial :
        21043
      1. Author :
        Ando, Hidenori; Kobayashi, Sakiko; Lila, Amr S Abu; Eldin, Noha Essam; Kato, Chihiro; Shimizu, Taro; Ukawa, Masami; Kawazoe, Kazuyoshi; Ishida, Tatsuhiro
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Journal of Controlled Release
      6. Products :
      7. Volume :
        220
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Malignant pleural mesothelioma;; Pemetrexed;; Liposomes;; Orthotopic tumor model;; Intrapleural administration; IVIS; IVIS BLI in vivo
      12. Abstract :
        Malignant pleural mesothelioma (MPM) is an aggressive cancer that proliferates in the pleural cavity. Pemetrexed (PMX) in combination with cisplatin is currently the approved standard care for MPM, but a dismal response rate persists. Recently, we prepared various liposomal PMX formulations using different lipid compositions and evaluated their in vitro cytotoxicity against human mesothelioma cells (MSTO-211 H). In the present study, we investigated the in vivo therapeutic effect of our liposomal PMX formulations using an orthotopic MPM tumor mouse model. PMX encapsulated within either cholesterol-containing (PMX/Chol CL) or cholesterol-free (PMX/Non-Chol CL) cationic liposome was intrapleurally injected into tumor-bearing mice. PMX encapsulated in cholesterol-free liposomes (PMX/Non-Chol CL) drastically inhibited the tumor growth in the pleural cavity, while free PMX and PMX encapsulated in cholesterol-containing liposomes (PMX/Chol CL) barely inhibited the tumor growth. The enhanced in vivo anti-tumor efficacy of PMX/Non-Chol CL was credited, on the one hand, for prolonging the retention of cationic liposomes in the pleural cavity via their electrostatic interaction with the negatively charged membranes of tumor cells, but on the other hand, it was charged with contributing to a higher drug release from the “fluid” liposomal membrane following intrapleural administration. This therapeutic strategy of direct intrapleural administration of liposomal PMX, along with the great advances in CL-guided therapeutics, might be a promising therapeutic approach to conquering the poor prognosis for MPM.
      13. URL :
        http://www.sciencedirect.com/science/article/pii/S0168365915301875
      14. Call Number :
        PKI @ user @ 10263
      15. Serial :
        21042
      1. Author :
        Andries, Oliwia; Mc Cafferty, Séan; De Smedt, Stefaan C.; Weiss, Ron; Sanders, Niek N.; Kitada, Tasuku
      2. Title :
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Journal of Controlled Release
      6. Products :
      7. Volume :
        217
      8. Issue :
        N/A
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        mRNA; Modified RNA; Nucleobase modifications; Antiviral innate immunity; Toll-like receptor; Gene therapy; IVIS; IVIS BLI in vivo
      12. Abstract :
        Messenger RNA as a therapeutic modality is becoming increasingly popular in the field of gene therapy. The realization that nucleobase modifications can greatly enhance the properties of mRNA by reducing the immunogenicity and increasing the stability of the RNA molecule (the Kariko paradigm) has been pivotal for this revolution. Here we find that mRNAs containing the N1-methylpseudouridine (m1Ψ) modification alone and/or in combination with 5-methylcytidine (m5C) outperformed the current state-of-the-art pseudouridine (Ψ) and/or m5C/Ψ-modified mRNA platform by providing up to ~ 44-fold (when comparing double modified mRNAs) or ~ 13-fold (when comparing single modified mRNAs) higher reporter gene expression upon transfection into cell lines or mice, respectively. We show that (m5C/)m1Ψ-modified mRNA resulted in reduced intracellular innate immunogenicity and improved cellular viability compared to (m5C/)Ψ-modified mRNA upon in vitro transfection. The enhanced capability of (m5C/)m1Ψ-modified mRNA to express proteins may at least partially be due to the increased ability of the mRNA to evade activation of endosomal Toll-like receptor 3 (TLR3) and downstream innate immune signaling. We believe that the (m5C/)m1Ψ-mRNA platform presented here may serve as a new standard in the field of modified mRNA-based therapeutics.
      13. URL :
        http://www.sciencedirect.com/science/article/pii/S0168365915300948
      14. Call Number :
        PKI @ user @ 10078
      15. Serial :
        21041
      1. Author :
        Aoki, Hiroyuki; Nojiri, Mayumi; Mukai, Rieko; Ito, Shinzaburo
      2. Title :
        Near-infrared absorbing polymer nano-particle as a sensitive contrast agent for photo-acoustic imaging
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        Nanoscale
      6. Products :
      7. Volume :
        7
      8. Issue :
        1
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        IVIS; IVIS FLI in vivo
      12. Abstract :
        Polymer nano-particles (PNPs) with a near-infrared (NIR) light absorption were prepared by the nano-emulsion method to develop contrast agents for photo-acoustic (PA) imaging. The PNP containing silicon naphthalocyanine showed a high absorption coefficient up to 1010 M−1 cm−1. This is comparable to plasmonic gold nano-particles, which have been studied as PA contrast agents. For the PNP larger than 100 nm, the enhancement of the PA signal was observed compared to the gold nano-particle with a similar absorption coefficient and size. In the case of the PNP, the heat by the light absorption is confined in the particle due to the low thermal diffusivity of polymer materials. We showed that the strong thermal confinement effect of PNP results in the enhancement of the efficiency of the PA signal generation and that the PA intensity can be enhanced by the increase of the Grüneisen parameter of the matrix polymer of PNP. The PA signal from the PNP of poly(methyl methacrylate) was 9-fold larger than that of gold nano-particles with the same absorption coefficient. We demonstrated that in the in vivo PA imaging the detection limit of PNP was of the order of 10−13 M. The NIR absorbing PNP will be a promising candidate of a sensitive contrast agent for PA imaging.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 8814
      15. Serial :
        21040
      1. Author :
        Aravalli, Rajagopal N; Talbot, Neil C; Steer, Clifford J
      2. Title :
        Gene expression profiling of MYC-driven tumor signatures in porcine liver stem cells by transcriptome sequencing
      3. Type :
        Journal Article
      4. Year :
        2015
      5. Publication :
        World journal of gastroenterology: WJG
      6. Products :
      7. Volume :
        21
      8. Issue :
        7
      9. Page Numbers :
        N/A
      10. Research Area :
        N/A
      11. Keywords :
        Hepatocellular carcinoma;; MYC;; Stem cells;; Gene expression;; RNA sequencing; IVIS; IVIS BLI in vitro
      12. Abstract :
        AIM: To identify the genes induced and regulated by the MYC protein in generating tumors from liver stem cells. METHODS: In this study, we have used an immortal porcine liver stem cell line, PICM-19, to study the role of c-MYC in hepatocarcinogenesis. PICM-19 cells were converted into cancer cells (PICM-19-CSCs) by overexpressing human MYC. To identify MYC-driven differential gene expression, transcriptome sequencing was carried out by RNA sequencing, and genes identified by this method were validated using real-time PCR. In vivo tumorigenicity studies were then conducted by injecting PICM-19-CSCs into the flanks of immunodeficient mice. RESULTS: Our results showed that MYC-overexpressing PICM-19 stem cells formed tumors in immunodeficient mice demonstrating that a single oncogene was sufficient to convert them into cancer cells (PICM-19- CSCs). By using comparative bioinformatics analyses, we have determined that > 1000 genes were differentially expressed between PICM-19 and PICM-19-CSCs. Gene ontology analysis further showed that the MYCinduced, altered gene expression was primarily associated with various cellular processes, such as metabolism, cell adhesion, growth and proliferation, cell cycle, inflammation and tumorigenesis. Interestingly, six genes expressed by PICM-19 cells (CDO1 , C22orf39 , DKK2 , ENPEP , GPX6 , SRPX2 ) were completely silenced after MYC-induction in PICM-19-CSCs, suggesting that the absence of these genes may be critical for inducing tumorigenesis. CONCLUSION: MYC-driven genes may serve as promising candidates for the development of hepatocellular carcinoma therapeutics that would not have deleterious effects on other cell types in the liver.
      13. URL :
        N/A
      14. Call Number :
        PKI @ user @ 9026
      15. Serial :
        21039
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