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Journal Cover Journal of Cellular Physiology
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   Hybrid Journal Hybrid journal (It can contain Open Access articles)
   ISSN (Print) 0021-9541 - ISSN (Online) 1097-4652
   Published by John Wiley and Sons Homepage  [1605 journals]
  • Cytoplasmic Localization of RUNX3 via Histone Deacetylase-Mediated SRC
           Expression in Oxidative-Stressed Colon Cancer Cells
    • Authors: Kyoung Ah Kang; Mei Jing Piao, Yea Seong Ryu, Young Hee Maeng, Jin Won Hyun
      Abstract: Runt domain transcription factor 3 (RUNX3) is a transcription factor that functions as a tumor suppressor. RUNX3 is frequently inactivated by epigenetic silencing or its protein mislocalization (cytoplasmic localization) in many cancer types. This study investigated whether oxidative stress induces redistribution of RUNX3 from the nucleus to the cytoplasm. The cytoplasmic localization of RUNX3 was associated with oxidative stress-induced RUNX3 phosphorylation at tyrosine residues via SRC activation. Moreover, oxidative stress increased expression of histone deacetylases (HDACs). RUNX3 phosphorylation and SRC expression induced by oxidative stress were inhibited by knockdown of HDAC1, restoring the nuclear localization of RUNX3 under oxidative stress. In conclusion, these results demonstrate that HDAC1- and SRC-mediated phosphorylation of RUNX3 induced by oxidative stress is associated with the cytoplasmic localization of RUNX3 and can lead to RUNX3 inactivation and carcinogenesis. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.RUNX3 cytoplasmic localization induced by oxidative stress in colon cancer cells. Oxidative stress induces HDAC1 expression and then activates SRC. Activated SRC induces expression of phospho-RUNX3. Phospho-RUNX3 is retained in the cytoplasm, leading to degradation of cytoplasmic RUNX3 by the proteasome system and finally resulting carcinogenesis.
      PubDate: 2017-02-21T05:31:21.597574-05:
      DOI: 10.1002/jcp.25746
  • Regulation of mitotic spindle orientation during epidermal stratification
    • Authors: Wei Xie; Jun Zhou
      Abstract: The epidermis is a stratified epithelium that serves as a barrier to infection from environmental pathogens and prevents water loss. Epidermal stratification is tightly controlled during embryogenesis. Progenitor cells in the developing epidermis undergo both symmetric and asymmetric cell divisions to balance the growth of the skin surface area against the generation of differentiated cell layers. Therefore, understanding the relationship between oriented divisions of progenitor cells and the development and stratification of the epidermis is of paramount importance in the field of skin biology and pathology. We provide here an integrated view of recent studies implicating that improper orientation of the mitotic spindle contributes to disorders associated with abnormal epidermal stratification and suggesting that spindle orientation could serve as a potential therapeutic target in skin diseases.We provide here an integrated view of recent studies implicating that improper orientation of the mitotic spindle contributes to disorders associated with abnormal epidermal stratification. We also suggest that spindle orientation could serve as a potential therapeutic target in skin diseases.
      PubDate: 2017-02-21T05:15:49.12907-05:0
      DOI: 10.1002/jcp.25750
  • LCN2 overexpression in bone enhances the hematopoietic compartment via
           modulation of the bone marrow microenvironment
    • Authors: Delfina Costa; Elisa Principi, Edoardo Lazzarini, Fiorella Descalzi, Ranieri Cancedda, Patrizio Castagnola, Sara Tavella
      Abstract: Lipocalin-2 (LCN2) is a member of the lipocalin family whose expression is modulated in several conditions, including cell differentiation, innate immunity, stress, and cancer. Although it is known that it is expressed in bone, its function in this tissue remains poorly studied. To this end, we took advantage of transgenic mice lines that expressed LCN2 driven by a bone specific type I collagen (LCN2-Tg). In the bone marrow (BM) of LCN2-Tg mice we observed an increased number of phenotypically long-term hematopoietic stem cells (LT-HSC) that also displayed a higher proliferation rate compared to wild-type controls (Wt). Furthermore, hematopoietic progenitor cells, obtained from LCN2-Tg BM showed an increased clonogenic capacity compared to those obtained from LCN2-Tg spleen, a higher concentration of serum erythropoietin and a higher number of mature erythrocytes in the peripheral blood of old LCN2-Tg animals compared to aged-matched wt. The findings of a combined increase in the BM of the LCN2-Tg mice of SDF-1, SCF, and TIMP-1 levels along with the reduction of both MMP-9 activity and cathepsin K concentration may explain the observed effects on the HSC compartment. This study shows that LCN2 overexpression in bones modifies the BM microenvironment via modulation of the expression of key secreted factors and cytokines, which in turn regulate the HSC niche behavior enhancing both HSC homing in young mice and erythrocytes production in older mice.The overexpression of LCN2 in bone stimulates the expansion of the HSC by inducing pro-retention factors and inhibiting pro-egress signals for hematopoietic stem cells.
      PubDate: 2017-02-21T05:10:50.678787-05:
      DOI: 10.1002/jcp.25755
  • Na+/H+ exchanger NHE1 and NHE2 have opposite effects on migration velocity
           in rat gastric surface cells
    • Authors: Anja Paehler vor der Nolte; Giriprakash Chodisetti, Zhenglin Yuan, Florian Busch, Brigitte Riederer, Min Luo, Yan Yu, Manoj B. Menon, Andreas Schneider, Renata Stripecke, Katerina Nikolovska, Sunil Yeruva, Ursula Seidler
      Abstract: Following superficial injury, neighbouring gastric epithelial cells close the wound by rapid cell migration, a process called epithelial restitution. Na+/H+ exchange (NHE) inhibitors interfere with restitution, but the role of the different NHE isoforms expressed in gastric pit cells has remained elusive. The role of the basolaterally expressed NHE1 (Slc9a1) and the presumably apically expressed NHE2 (Slc9a2) in epithelial restitution was investigated in the nontransformed rat gastric surface cell line RGM1. Migration velocity was assessed by loading the cells with the fluorescent dye DiR and following closure of an experimental wound over time. Since RGM1 cells expressed very low NHE2 mRNA and have low transport activity, NHE2 was introduced by lentiviral gene transfer. In medium with pH 7.4, RGM1 cells displayed slow wound healing even in the absence of growth factors and independently of NHE activity. Growth factors accelerated wound healing in a partly NHE1-dependent fashion. Preincubation with acidic pH 7.1 stimulated restitution in a NHE1-dependent fashion. When pH 7.1 was maintained during the restitution period, migratory speed was reduced to ∼10% of the speed at pH 7,4, and the residual restitution was further inhibited by NHE1 inhibition. Lentiviral NHE2 expression increased the steady-state pHi and reduced the restitution velocity after low pH preincubation, which was reversible by pharmacological NHE2 inhibition. The results demonstrate that in RGM1 cells, migratory velocity is increased by NHE1 activation, while NHE2 activity inhibit this process. A differential activation of NHE1 and NHE2 may therefore, play a role in the initiation and completion of the epithelial restitution process.Genetic deletion of the Na+/H+ exchanger NHE2 leads to compromised wound healing in the gastrointestinal tract of mice. We studied the roles of NHE1 and NHE2 in gastrointestinal wound healing in vitro, and while NHE1 activity supported wound healing velocity, NHE2/GFP-overexpression dramatically reduced wound healing velocity compared to GFP expression alone. This effect was abolished by a NHE2- but not a NHE1 inhibitory concentration of HOE642. Thus, NHE1 and NHE2 may be important for the initiation and the resolution of gastrointestinal wound healing.
      PubDate: 2017-02-21T05:05:58.470701-05:
      DOI: 10.1002/jcp.25758
  • Activin-SMAD signaling is required for maintenance of porcine iPS cell
           self-renewal through upregulation of NANOG and OCT4 expression
    • Authors: Fan Yang; Ning Wang, Yaxian Wang, Tong Yu, Huayan Wang
      Abstract: Porcine induced pluripotent stem cells (piPSCs) retain the enormous potential for farm animal reproduction and translational medicine, and have been reported by many laboratories worldwide. Some piPSC lines were bFGF-dependence and showed mouse EpiSC-like morphology; other lines were LIF-dependence and showed mouse ESC-like morphology. Metastable state of piPSC line that required both LIF and bFGF was also reported. Because bona fide pig embryonic stem cells were not available, uncovering piPSC state-specific regulatory circuitries was the most important task. In this study, we explored the function of Activin-SMAD signaling pathway and its downstream activated target genes in piPSCs. Transcriptome analysis showed that genes involved in Activin-SMAD signaling pathway were evidently activated during porcine somatic cell reprogramming, regardless piPSCs were LIF- or bFGF-dependent. Addition of Activin A and overexpression of SMAD2/3 significantly promoted expressions of porcine NANOG and OCT4, whereas inhibition of Activin-SMAD signaling by SB431542 and SMAD7 reduced NANOG and OCT4 expressions, and induced piPSCs differentiation exiting from pluripotent state. Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.Our data demonstrate that activation of Activin-SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self-renewal in porcine pluripotent stem cells.
      PubDate: 2017-02-21T04:40:33.007513-05:
      DOI: 10.1002/jcp.25747
  • Glucose Adsorption to Chitosan Membranes Increases Proliferation of Human
           Chondrocyte via Mammalian Target of Rapamycin Complex 1 and Sterol
           Regulatory Element-binding Protein-1 Signaling
    • Authors: Shun-Fu Chang; Kuo-Chin Huang, Chin-Chang Cheng, Yu-Ping Su, Ko-Chao Lee, Cheng-Nan Chen, Hsin-I Chang
      Abstract: BackgroundOsteoarthritis (OA) is currently still an irreversible degenerative disease of the articular cartilage. Recent, dextrose (D-glucose) intraarticular injection prolotherapy for OA patients has been reported to benefit the chondrogenic stimulation of damaged cartilage. However, the detailed mechanism of glucose's effect on cartilage repair remains unclear. Chitosan, a naturally derived polysaccharide, has recently been investigated as a surgical or dental dressing to control breeding. Therefore, in this study, glucose was adsorbed to chitosan membranes (CTS-Glc), and the study aimed to investigate whether CTS-Glc complex membranes could regulate the proliferation of human OA chondrocytes and to explore the underlying mechanism.MethodsHuman OA and SW1353 chondrocytes were used in this study. The experiments involving the transfection of cells used SW1353 chondrocytes. A specific inhibitor and siRNAs were used to investigate the mechanism underlying the CTS-Glc-regulated proliferation of human chondrocytes.ResultsWe found that CTS-Glc significantly increased the proliferation of both human OA and SW1353 chondrocytes comparable to glucose- or chitosan-only stimulation. The role of mammalian target of rapamycin complex 1 (mTORC1) signaling, including mTOR, raptor, and S6k proteins, has been demonstrated in the regulation of CTS-Glc-increased human chondrocyte proliferation. mTORC1 signaling increased the expression levels of maturated SREBP-1 and FASN and then induced the expressions of cell cycle regulators, i.e., cyclin D, cyclin-dependent kinase-4 and -6 in human chondrocytes.ConclusionsThis study elucidates the detailed mechanism behind the effect of CTS-Glc complex membranes in promoting chondrocyte proliferation and proposes a possible clinical application of the CTS-Glc complex in the dextrose intraarticular injection of OA prolotherapy in the future to attenuate the pain and discomfort of OA patients. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-20T07:51:23.531003-05:
      DOI: 10.1002/jcp.25869
  • MAPK3/1 Participates in the Activation of Primordial Follicles through
           mTORC1-KITL Signaling
    • Authors: Yu Zhao; Yu Zhang, Jia Li, Nana Zheng, Xiaoting Xu, Jing Yang, Guoliang Xia, Meijia Zhang
      Abstract: The majority of ovarian primordial follicles are preserved in a dormant state to maintain the female reproductive lifespan, and only a few primordial follicles are activated to enter the growing follicle pool in each wave. Recent studies have shown that primordial follicular activation depends on mammalian target of rapamycin complex 1 (mTORC1)-KIT ligand (KITL) signaling in pre-granulosa cells and its receptor (KIT)-phosphoinositol 3 kinase (PI3K) signaling in oocytes. However, the upstream regulator of mTORC1 signaling is unclear. The results of the present study showed that the phosphorylated mitogen-activated protein kinase3/1 (MAPK3/1) protein is expressed in some primordial follicles and all growing follicles. Culture of 3 days post-parturition (dpp) ovaries with the MAPK3/1 signaling inhibitor U0126 significantly reduced the number of activated follicles and was accompanied by dramatically reduced granulosa cell proliferation and increased oocyte apoptosis. Western blot and immunofluorescence analyses showed that U0126 significantly decreased the phosphorylation levels of Tsc2, S6K1, and rpS6 and the expression of KITL, indicating that U0126 inhibits mTORC1-KITL signaling. Furthermore, U0126 decreased the phosphorylation levels of Akt, resulting in a decreased number of oocytes with Foxo3 nuclear export. To further investigate MAPK3/1 signaling in primordial follicle activation, we used phosphatase and tensin homolog deleted on chromosome 10 (PTEN) inhibitor bpV(HOpic) to promote primordial follicle activation. In this model, U0126 also inhibited the activation of primordial follicles and mTORC1 signaling. Thus, these results suggest that MAPK3/1 participates in primordial follicle activation through mTORC1-KITL signaling. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-20T07:45:26.212931-05:
      DOI: 10.1002/jcp.25868
  • Palmitate Induces Mitochondrial Superoxide Generation and Activates AMPK
           in Podocytes
    • Authors: Eugene Lee; Jin Choi, Hyun Soon Lee
      Abstract: Studies have shown that high levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 diabetes. Palmitic acid (PA) is the predominant circulating saturated FFA, yet its role in the pathogenesis of diabetic nephropathy (DN) is not clear. Recently, one study suggested that mitochondrial superoxide production is related to AMP-activated protein kinase (AMPK) activity in diabetic mice kidneys. To elucidate the link between PA and oxidative stress and AMPK activity in DN, we compared the cultured murine podocytes exposed to PA and oleic acid (OA). Incubation of cells with 250 µM PA or OA induced a translocation of CD36, a fatty acid transport protein, with intracellular lipid accumulation. PA, but not OA, induced mitochondrial superoxide and hydrogen peroxide (H2O2) generation in podocytes, as shown by enhanced fluorescence of MitoSOX Red and dichlorofluorescein (DCF), respectively. Costimulation of PA-treated cells with the H2O2 scavenger catalase abolished the PA-induced DCF fluorescence. Only PA induced mitochondrial damage as shown by electron microscopy. The AMPK activity was determined by immunobloting, measuring the ratio of phosphorylated AMPK (p-AMPK) to total AMPK. Only PA significantly increased the p-AMPK levels compared with controls. Addition of catalase to PA-treated cells did not affect the PA-stimulated p-AMPK levels. Collectively, our results indicate that PA induces mitochondrial superoxide and H2O2 generation in cultured podocytes, which may not be directly linked to AMPK activation. Given that, PA seems to play an important role in the pathogenesis of DN through lipotoxicity initiated by mitochondrial superoxide overproduction. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T10:00:27.672401-05:
      DOI: 10.1002/jcp.25867
  • Mitochondria-targeted Esculetin Inhibits PAI-1 Levels by Modulating STAT3
           Activation and miR-19b via SIRT3: Role in Acute Coronary Artery Syndrome
    • Authors: Sujana Katta; Santosh Karnewar, Devayani Panuganti, Mahesh Kumar Jerald, Sastry B K, Srigiridhar Kotamraju
      Abstract: In this study, we explored the microRNAs responsible for the regulation of PAI-1 during LPS-stimulated inflammation in human aortic endothelial cells, subsequently studied the effect of a newly synthesized mitochondria-targeted esculetin (Mito-Esc) that was shown for its anti-atherosclerotic potential in modulating PAI-1 levels and its targeted miRs during angiotensin-II-induced atherosclerosis in ApoE-/- mice. LPS-stimulated PAI-1 was accompanied with an upregulation of miR-19b and down-regulation of miR-30c. These effects of LPS on PAI-1 were reversed in the presence of both parent esculetin and Mito-Esc. However, the effect of Mito-Esc was more pronounced in the regulation of PAI-1. In addition, LPS-stimulated PAI-1 expression was significantly decreased in cells treated with Anti-miR-19b, thereby suggesting that miR-19b co-expression plays a key role in PAI-1 regulation. The results also show that incubation of cells with Stattic, an inhibitor of STAT-3, inhibited LPS-stimulated PAI-1 expression. Interestingly, knockdown of SIRT3, a mitochondrial biogenetic marker, enhanced PAI-1 levels via modulation of miR-19b and -30c. Mito-Esc treatment significantly inhibited Ang-II-induced PAI-1, possibly via altering miR-19b and 30c in ApoE-/- mice. The association between PAI-1, miR-19b and -30c were further confirmed in plasma and microparticles isolated from patients suffering from acute coronary syndrome of various degrees. Taken together, LPS-induced PAI-1 involves co-expression of miR-19b and down regulation of miR-30c and Mito-Esc treatment by modulating miR-19b and miR-30c through SIRT3 activation, inhibits PAI-1 levels that, in part, contribute to its anti-atherosclerotic effects. Moreover, there exists a strong positive correlation between miR-19b and PAI-1 in patients suffering from ST-elevated myocardial infarction. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:20:37.618848-05:
      DOI: 10.1002/jcp.25865
  • Transforming Growth Factor β Induces Bone Marrow Mesenchymal Stem Cell
           Migration via Noncanonical Signals and N-cadherin
    • Authors: Maria Jose Dubon; Jinyeong Yu, Sanghyuk Choi, Ki-Sook Park
      Abstract: Transforming growth factor-beta (TGF-β) induces the migration and mobilization of bone marrow-derived mesenchymal stem cells (BM-MSCs) to maintain bone homeostasis during bone remodeling and facilitate the repair of peripheral tissues. Although many studies have reported the mechanisms through which TGF-β mediates the migration of various types of cells, including cancer cells, the intrinsic cellular mechanisms underlying cellular migration and mobilization of BM-MSCs mediated by TGF-β are unclear. In this study, we showed that TGF-β activated noncanonical signaling molecules, such as Akt, extracellular signal-regulated kinase 1/2 (ERK1/2), focal adhesion kinase (FAK), and p38, via TGF-β type I receptor in human BM-MSCs and murine BM-MSC-like ST2 cells. Inhibition of Rac1 by NSC23766 and Src by PP2 resulted in impaired TGF-β-mediated migration. These results suggested that the Smad-independent, noncanonical signals activated by TGF-β were necessary for migration. We also showed that N-cadherin-dependent intercellular interactions were required for TGF-β-mediated migration using functional inhibition of N-cadherin with EDTA treatment and a neutralizing antibody (GC-4 antibody) or siRNA-mediated knockdown of N-cadherin. However, N-cadherin knockdown did not affect the global activation of noncanonical signals in response to TGF-β. Therefore, these results suggested that the migration of BM-MSCs in response to TGF-β was mediated through N-cadherin and noncanonical TGF-β signals. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:20:33.723598-05:
      DOI: 10.1002/jcp.25863
  • Promotion of Cell Growth and Adhesion of a Peptide Hydrogel Scaffold via
           mTOR/Cadherin Signaling
    • Authors: Guojun Wei; Liping Wang, Daming Dong, Zhaowei Teng, Zuowei Shi, Kaifu Wang, Gang An, Ying Guan, Bo Han, Meng Yao, Cory J. Xian
      Abstract: Understanding neurite outgrowth, orientation, and migration is important for the design of biomaterials that interface with the neural tissue. However, the molecular signaling alternations have not been well elucidated to explain the impact of hydrogels on cell morphology. In our previous studies, a silk fibroin peptide (SF16) hydrogel was found to be an effective matrix for the viability, morphology and proliferation of PC12 rat pheocrhomocytoma cells. We found that PC12 cells in the peptide hydrogel exhibited adhesive morphology compared to those cultured in agarose or collagen. Moreover, we identified that cell adhesion molecules (E- and N-cadherin) controlled by mTOR signaling were highly induced in PC12 cells cultured in the SF16 peptide hydrogel. Our findings suggest that the SF16 peptide might be suitable to be a cell-adhesion material in cell culture or tissue engineering, and mTOR/cadherin signaling is required for the cell adhesion in the SF16-peptide hydrogel. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:20:25.809879-05:
      DOI: 10.1002/jcp.25864
  • Chrysin Induces Death of Prostate Cancer Cells by Inducing ROS and ER
    • Authors: Soomin Ryu; Whasun Lim, Fuller W. Bazer, Gwonhwa Song
      Abstract: Chrysin is a natural flavone found in numerous plant extracts, honey and propolis that has multiple biological activities including anti-cancer effects. Understanding of biological mechanisms mediated in response to chrysin in cancerous cells may provide novel insight into chemotherapeutic approaches with reduced side effects in cancers. In the present study, we investigated functional roles of chrysin in progression of prostate cancer cells using DU145 and PC-3 cell lines. The results showed that chrysin induced apoptosis of cells evidenced by DNA fragmentation and increasing the population of both DU145 and PC-3 cells in the sub-G1 phase of the cell cycle. In addition, chrysin reduced expression of proliferating cell nuclear antigen in the prostate cancer cell lines compared to untreated prostate cancer cells. Moreover, chrysin induced loss of mitochondria membrane potential (MMP), while increasing production of reactive oxygen species (ROS) and lipid peroxidation in a dose-dependent manner. Also, it induced endoplasmic reticulum (ER) stress through activation of unfolded protein response (UPR) proteins including PRKR-like ER kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), and 78 kDa glucose-regulated protein (GRP78) in DU145 and PC-3 cells. The chrysin-mediated intracellular signaling pathways suppressed phosphoinositide 3-kinase (PI3K) and the abundance of AKT, P70S6K, S6 and P90RSK proteins, but stimulated mitogen-activated protein kinases (MAPK) and activation of ERK1/2 and P38 proteins in the prostate cancer cells. Collectively, these results indicate that chrysin initiates cell death through induction of mitochondrial-mediated apoptosis and ER stress, and regulation of signaling pathways responsible for proliferation of prostate cancer cells. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:15:30.691736-05:
      DOI: 10.1002/jcp.25861
  • Estrogens and Androgens Inhibit Association of RANKL with the
           Pre-osteoblast Membrane through Post-translational Mechanisms
    • Authors: Anthony Martin; Jiali Yu, Jian Xiong, Aysha B. Khalid, Benita Katzenellenbogen, Sung Hoon Kim, John A. Katzenellenbogen, Suchinda Malaivijitnond, Yankel Gabet, Susan A. Krum, Baruch Frenkel
      Abstract: We have recently demonstrated that RUNX2 promoted, and 17-β-Estradiol (E2) diminished, association of RANKL with the cell membrane in pre-osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL and transiently transfected GFP-RANKL with the pre-osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane-associated RANKL was accompanied with marked suppression of osteoclast differentiation from co-cultured pre-osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre-osteoblast conditioned media. A marked decrease in membrane-associated RANKL was observed after 30 minutes of either E2 or DHT treatment, and near-complete inhibition was observed by 1 hour, suggesting that the diminution of RANKL membrane association was mediated through non-genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2-mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast-driven osteoclastogenesis through non-genomic, MMP-mediated RANKL dissociation from the cell membrane. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-18T05:15:27.41734-05:0
      DOI: 10.1002/jcp.25862
  • Mesenchyme Homeobox 2 Enhances Migration of Endothelial Colony Forming
           Cells Exposed to Intrauterine Diabetes Mellitus
    • Authors: Cassandra R. Gohn; Emily K. Blue, BreAnn M. Sheehan, Kaela M. Varberg, Laura S. Haneline
      Abstract: Diabetes mellitus (DM) during pregnancy has long-lasting implications for the fetus, including cardiovascular morbidity. Previously, we showed that endothelial colony forming cells (ECFCs) from DM human pregnancies have decreased vasculogenic potential. Here, we evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). In human umbilical vein endothelial cells, MEOX2 upregulates cyclin-dependent kinase inhibitor expression, resulting in increased senescence and decreased proliferation. We hypothesized that dysregulated MEOX2 expression in neonatal ECFCs from DM pregnancies decreases network formation through increased senescence and altered cell cycle progression. Our studies show that nuclear MEOX2 is increased in ECFCs from DM pregnancies. To determine if MEOX2 is sufficient and/or required to induce impaired network formation, MEOX2 was overexpressed and depleted in ECFCs from control and DM pregnancies, respectively. Surprisingly, MEOX2 overexpression in control ECFCs resulted in increased network formation, altered cell cycle progression, and increased senescence. In contrast, MEOX2 knockdown in ECFCs from DM pregnancies led to decreased network formation, while cell cycle progression and senescence were unaffected. Importantly, migration studies demonstrated that MEOX2 overexpression increased migration, while MEOX2 knockdown decreased migration. Taken together, these data suggest that altered migration may be mediating the impaired vasculogenesis of ECFCs from DM pregnancies. While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Endothelial colony forming cells (ECFCs) from diabetic human pregnancies have decreased vasculogenic potential both in vitro and in vivo. The goal of this study was to evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment.
      PubDate: 2017-02-16T10:25:26.580862-05:
      DOI: 10.1002/jcp.25734
  • The Role of Cardiac Myosin Binding Protein C3 in Hypertrophic
           Cardiomyopathy-Progress and Novel Therapeutic Opportunities
    • Authors: Iman A. Mohamed; Navaneethakrishnan T. Krishnamoorthy, Gheyath K. Nasrallah, Sahar I. Da'as
      Abstract: Hypertrophic cardiomyopathy (HCM) is a common autosomal dominant genetic cardiovascular disorder marked by genetic and phenotypic heterogeneity. Mutations in the gene encodes the cardiac myosin-binding protein C, cMYBPC3 is amongst the various sarcomeric genes that are associated with HCM. These mutations produce mutated mRNAs and truncated cMyBP-C proteins. In this review, we will discuss the implications and molecular mechanisms involved in MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of HMC. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.This review discusses the implications and the and molecular mechanisms of MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of Hypertrophic cardiomyopathy.
      PubDate: 2017-02-16T10:15:27.995734-05:
      DOI: 10.1002/jcp.25639
  • Identification of MSX1 and DCLK1 as mRNA Biomarkers for Colorectal Cancer
           Detection Through DNA Methylation Information
    • Authors: Ai-Jun Sun; Hai-Bo Gao, Gao Liu, Heng-Fa Ge, Zun-Ping Ke, Sen Li
      Abstract: Colorectal cancer is the second most deadly malignancy in the United States. However, the currently screening options had their limitation. Novel biomarkers for colorectal cancer detections are necessary to reduce the mortality. The clinical information, mRNA expression levels and DNA methylation information of colorectal cancer were downloaded from TCGA. The patients were separated into training group and testing group based on their platforms for DNA methylation. Beta values of DNA methylation from tumor tissues and normal tissues were utilized to figure out the position that were differentially methylated. The expression levels of mRNA of thirteen genes, whose CpG islands were differentially methylated, were extracted from the RNA-Seq results from TCGA. The probabilities whether the mRNA was differentially expressed between tumor and normal samples were calculated using Student's t-test. Logistic regression and decision tree were built for cancer detection and their performances were evaluated by the area under the curve (AUC). Twenty-four genomic locations were differentially methylated, which could be mapped to eleven genes. Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies. J. Cell. Physiol. 9999: 1–6, 2016. © 2016 Wiley Periodicals, Inc.Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies.
      PubDate: 2017-02-16T10:10:26.023949-05:
      DOI: 10.1002/jcp.25733
  • MicroRNA-125a-5p Is a Downstream Effector of Sorafenib in Its
           Antiproliferative Activity Toward Human Hepatocellular Carcinoma Cells
    • Authors: Nicoletta Potenza; Nicola Mosca, Silvia Zappavigna, Filomena Castiello, Marta Panella, Carmela Ferri, Daniela Vanacore, Antonio Giordano, Paola Stiuso, Michele Caraglia, Aniello Russo
      Abstract: Sorafenib is an antitumor drug for treatment of advanced hepatocellular carcinoma (HCC). It acts as a multikinase inhibitor suppressing cell proliferation and angiogenesis. Human microRNA-125a-5p (miR-125a) is endowed with similar activities and is frequently downregulated in HCC. Looking for a potential microRNA-based mechanism of action of the drug, we found that sorafenib increases cellular expression of miR-125a in cultured HuH-7 and HepG2 HCC cells. Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin-7, a NAD(+)-dependent deacetylase, and p21/p27-dependent cell cycle arrest in G1. Later, recruitment of miR-125a in the antiproliferative activity of sorafenib was inquired by modulating its expression in combination with the drug treatment. This analysis showed that intracellular delivery of miR-125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR-125a inhibitor could counteract it. Finally, evaluation of other oncogenic targets of miR-125a revealed its ability to interfere with the expression of matrix metalloproteinase-11, Zbtb7a proto-oncogene, and c-Raf, possibly contributing to the antiproliferative activity of the drug. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc.Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin-7, a NAD(+)-dependent deacetylase, and p21/p27-dependent cell cycle arrest in G1. Intracellular delivery of miR-125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR-125a inhibitor could counteract it. miR-125a interfered with the expression of matrix metalloproteinase-11, Zbtb7a proto-oncogene, and c-Raf, possibly contributing to the antiproliferative activity of the drug.
      PubDate: 2017-02-16T10:05:25.695604-05:
      DOI: 10.1002/jcp.25744
  • Dact1, a Wnt-Pathway Inhibitor, Mediates Human Mesangial Cell
           TGF-β1-Induced Apoptosis
    • Authors: Daniele Pereira Jardim; Paula Cristina Eiras Poço, Alexandre Holthausen Campos
      Abstract: Chronic kidney disease (CKD) is a worldwide public health problem that affects millions of men and women of all ages and racial groups. Loss of mesangial cells (MC) represents an early common feature in the pathogenesis of CKD. Transforming growth factor-β1 (TGF-β1) is a key inducer of kidney damage and triggers several pathological changes in renal cells, notably MC apoptosis. However, the mechanism of MC apoptosis induced by TGF-β1 remains elusive. Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled-binding antagonist of β-catenin 1 (Dact1) gene is upregulated by TGF-β1, inducing MC apoptosis. We also show that the inhibitory effect of Dact1 and TGF-β1 on the transcriptional activation of the pro-survival Wnt pathway is the mechanism of death induction. In addition, Dact1 mRNA/protein levels are increased in kidney remnants from 5/6 nephrectomized rats and strongly correlate with TGF-β1 expression. Together, our results point to Dact1 as a novel element controlling MC survival that is causally related to CKD progression. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled-binding antagonist of β-catenin 1 (Dact1) gene is upregulated by TGF-β1, inducing MC apoptosis.
      PubDate: 2017-02-16T09:46:07.825522-05:
      DOI: 10.1002/jcp.25636
  • Proteomics Applications in Dental Derived Stem Cells
    • Authors: Jie Li; Weidong Tian, Jinlin Song
      Abstract: At present, the existence of a variety of dental derived stem cells has been documented. These cells displayed promising clinical application potential not only for teeth and its surrounding tissue regeneration, but also for other tissues, such as nerve and bone regeneration. Proteomics is an unbiased, global informatics tool that provides information on all protein expression levels as well as post-translational modification in cells or tissues and is applicable to dental derived stem cells research. Over the last decade, considerable progress has been made to study the global proteome, secrotome, and membrane proteome of dental derived stem cells. Here, we present an overview of the proteomics studies in the context of stem cell research. Particular attention is given to dental derived stem cell types as well as current challenges and opportunities. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.(1) Dental derived stem cells displayed promising clinical application potential not only for teeth and its surrounding tissue regeneration, but also for other tissues regeneration. (2) Here, we present an overview of the proteomics studies in the context of dental derived stem cell research. (3) Particular attention is given to dental derived stem cell types as well as current challenges and opportunities.
      PubDate: 2017-02-16T09:40:25.198627-05:
      DOI: 10.1002/jcp.25667
  • Tight Junction Protein Abundance and Apoptosis During Involution of Rat
           Mammary Glands
    • Authors: Claire V. C. Phyn; Kerst Stelwagen, Stephen R. Davis, Christopher D. McMahon, Joanne M. Dobson, Kuljeet Singh
      Abstract: To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement were induced by teat-sealing with an adhesive for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. In non-sealed control glands, histological analysis confirmed a lactating phenotype, indicating suckling by pups throughout the experiment. In contrast, alveoli of teat-sealed glands were distended within 6 h, with maximal luminal size observed by 12 h of non-suckling. By 18 h following teat-sealing, an involuting phenotype was observed, indicated by alveolar lumina engorged with milk vesicles and increased leukocytes. Relative to non-sealed glands, mammary apoptosis was increased in engorged glands 18 h following teat-sealing. The abundance of ZO-1 and occludin proteins was decreased in engorged glands by 12 and 18 h, respectively, following teat-sealing. In contrast, the claudin-1 22 kDa band was increased by 6 h and peaked at 12–18 h, whereas the 28 kDa band declined by 36 h, relative to controls. There were no temporal changes in ZO-1, occludin, and claudin-1 22 kDa proteins within control glands, although there were minor differences in claudin-1 28 kDa. These data indicate that intramammary milk accumulation due to cessation of milk removal is associated with mammary apoptosis. The apoptotic event is preceded by a rapid loss of abundance of ZO-1, occludin and an initial increase in claudin-1. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non-suckled glands. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement was induced by teat-sealing for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non-suckled glands.
      PubDate: 2017-02-16T09:30:26.601031-05:
      DOI: 10.1002/jcp.25591
  • Protective Role of GPER Agonist G-1 on Cardiotoxicity Induced by
    • Authors: Ernestina M. De Francesco; Carmine Rocca, Francesco Scavello, Daniela Amelio, Teresa Pasqua, Damiano C. Rigiracciolo, Andrea Scarpelli, Silvia Avino, Francesca Cirillo, Nicola Amodio, Maria C. Cerra, Marcello Maggiolini, Tommaso Angelone
      Abstract: The use of Doxorubicin (Dox), a frontline drug for many cancers, is often complicated by dose-limiting cardiotoxicity in approximately 20% of patients. The G-protein estrogen receptor GPER/GPR30 mediates estrogen action as the cardioprotection under certain stressful conditions. For instance, GPER activation by the selective agonist G-1 reduced myocardial inflammation, improved immunosuppression, triggered pro-survival signaling cascades, improved myocardial mechanical performance, and reduced infarct size after ischemia/reperfusion (I/R) injury. Hence, we evaluated whether ligand-activated GPER may exert cardioprotection in male rats chronically treated with Dox. 1 week of G-1 (50 μg/kg/day) intraperitoneal administration mitigated Dox (3 mg/kg/day) adverse effects, as revealed by reduced TNF-α, IL-1β, LDH, and ROS levels. Western blotting analysis of cardiac homogenates indicated that G-1 prevents the increase in p-c-jun, BAX, CTGF, iNOS, and COX2 expression induced by Dox. Moreover, the activation of GPER rescued the inhibitory action elicited by Dox on the expression of BCL2, pERK, and pAKT. TUNEL assay indicated that GPER activation may also attenuate the cardiomyocyte apoptosis upon Dox exposure. Using ex vivo Langendorff perfused heart technique, we also found an increased systolic recovery and a reduction of both infarct size and LDH levels in rats treated with G-1 in combination with Dox respect to animals treated with Dox alone. Accordingly, the beneficial effects induced by G-1 were abrogated in the presence of the GPER selective antagonist G15. These data suggest that GPER activation mitigates Dox-induced cardiotoxicity, thus proposing GPER as a novel pharmacological target to limit the detrimental cardiac effects of Dox treatment. J. Cell. Physiol. 9999: 1–10 , 2016. © 2016 Wiley Periodicals, Inc.1. The potential beneficial effects of GPER activation in Doxorubicin (Dox)-induced cardiotoxicity was evaluated. 2. In male rat heart, the treatment with the selective GPER ligand G-1 prevented the increase in TNF-α, IL-1β, and LDH plasma levels induced by Dox. In Langendorff perfused rat heart, G-1 also increased the systolic recovery at the end of ischemia and reduced infarct size and LDH levels triggered by Dox. Moreover, G-1 attenuated the Dox-induced myocyte apoptosis, as well as the inhibition of pro-survival signaling cascades like ERK1/2 and AKT. Next, G-1 counteracted the down-regulation of BCL2 as well as the up-regulation of apoptotic, inflammatory and fibrotic targets, such as BAX, iNOS, COX2, and CTGF induced by Dox. 3. Our study paves the way for further analyzing the role of GPER as pharmacological target in novel therapeutic strategies aimed to prevent the cardiotoxicity exerted by Dox.
      PubDate: 2017-02-16T09:20:30.356957-05:
      DOI: 10.1002/jcp.25585
  • Postnatal Hyperplasic Effects of ActRIIB Blockade in a Severely Dystrophic
    • Authors: Cory Nielsen; Ross M. Potter, Christopher Borowy, Kimberly Jacinto, Ravi Kumar, C. George Carlson
      Abstract: The efficacy of two ActRIIB ligand-trapping agents (RAP-031 and RAP-435) in treating muscular dystrophy was examined by determining their morphological effects on the severely dystrophic triangularis sterni (TS) muscle of the mdx mouse, a model for Duchenne muscular dystrophy. These agents trap all endogenous ligands to the ActRIIB receptor and thereby block myostatin signaling in a highly selective manner. Short-term (1 month) and long-term (3 months) in vivo treatment of 1-month-old mdx mice increased myonuclei and fiber cross section (FCS) density but did not alter individual fiber size. Vehicle-treated mdx mice exhibited age-dependent increases in myonuclei and FCS density, and age-dependent reductions in centronucleation that were each enhanced by treatment with RAP-435. Distributions of FCS area (FCSA) in the mdx TS were 90% identical to those from untreated age-matched nondystrophic mice and were unaltered by the substantial fiber hyperplasia observed with age and RAP-435 treatment. These results were inconsistent with injury-induced fiber regeneration which produces altered FCSA distributions characterized by a distinct class of smaller regenerated fibers. Nondystrophic mice exhibited a constant postnatal density of fiber cross sections and myonuclei, and RAP-435 treatment of nondystrophic mice increased TS mean FCSA but had no effects on myonuclei or FCS density. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration. J. Cell. Physiol. 9999: 1–20, 2016. © 2016 Wiley Periodicals, Inc.We examined the efficacy of two ActRIIB ligand-trapping agents in altering the morphology of a severely dystrophic muscle in the mdx mouse. The results indicate that severely dystrophic muscle exhibits monotonic postnatal increases in myonuclei and fiber density that are not associated with alterations in the distribution of fiber cross-sectional areas. Long-term treatment with an ActRIIB ligand-trapping agent substantially enhanced this postnatal hyperplasia and reduced centronucleation. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration.
      PubDate: 2017-02-16T09:15:56.755686-05:
      DOI: 10.1002/jcp.25694
  • Role of Wnt5a in the Pathogenesis of Inflammatory Diseases
    • Authors: Mehran Pashirzad; Mojtaba Shafiee, Farzad Rahmani, Reihane Behnam-Rassouli, Fatemeh Hoseinkhani, Mikhail Ryzhikov, Maryam Moradi Binabaj, Mohammad Reza Parizadeh, Amir Avan, Seyed Mahdi Hassanian
      Abstract: Wnt5a initiates pro-inflammatory responses through activation of non-canonical Wnt signaling pathway. Pro-inflammatory functions of Wnt5a trigger pro-inflammatory signaling cascades and increase secretion of pro-inflammatory cytokines and chemokines. Wnt5a as a potent signaling molecule is strongly implicated in a number of diseases including cancer, diabetes, metabolic disorders, and of special interest in this review, inflammatory diseases. This review summarizes the role of Wnt5a in the pathogenesis of inflammatory diseases including atherosclerosis, rheumatoid arthritis, psoriasis vulgaris and sepsis, promoting greater understanding, and clinical management of these diseases. J. Cell. Physiol. 9999: 1–6, 2017. © 2016 Wiley Periodicals, Inc.Wnt5a as a potent signaling molecule is strongly implicated in a number of diseases including cancer, diabetes, metabolic disorders, and of special interest in this review, inflammatory diseases. This review summarizes the role of Wnt5a in the pathogenesis of inflammatory diseases including atherosclerosis, rheumatoid arthritis, psoriasis vulgaris and sepsis, promoting greater understanding and clinical management of these diseases.
      PubDate: 2017-02-16T09:10:27.206473-05:
      DOI: 10.1002/jcp.25687
  • A Perspective on Malignancy in the Marrow
    • Authors: Michaela R. Reagan; Jane Lian, Clifford J. Rosen, Gary Stein
      Abstract: Malignancies that grow in the bone marrow cause a cascade of devastating consequences and are almost always fatal. For researchers to make significant headway in finding cures and treatments for these tumors and the subsequent devastation of cancer-induced bone disease, novel strategies and creative solutions will have to be considered. Great progress has been made in treating hematologic and sold tumor malignancies that ultimately affect the bone marrow, although it is also obvious that multi-disciplinary teams are required to ultimately win the war on cancers that affect the bone. In this perspective, we review recent advances in the identification of molecular and cellular targets in the bone marrow niche. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-16T06:50:33.021627-05:
      DOI: 10.1002/jcp.25860
  • Modulating Tumor Hypoxia by Nanomedicine for Effective Cancer Therapy
    • Authors: Rana Jahanban-Esfahlan; Miguel de la Guardia, Delshad Ahmadi, Bahman Yousefi
      Abstract: Hypoxia, a characteristic feature of tumors, is indispensable to tumor angiogenesis, metastasis, and multi drug resistance. Hypoxic avascular regions, deeply embedded inside the tumors significantly hinder delivery of therapeutic agents. The low oxygen tension results in resistance to the current applied anti-cancer therapeutics including radiotherapy, chemotherapy and photodynamic therapy, the efficacy of which is firmly tied to the level of tumor oxygen supply. However, emerging data indicate that nanocarriers/nanodrugs can offer substantial benefits to improve the efficacy of current therapeutics, through modulation of tumor hypoxia. This review aims to introduce the most recent advances made in nanocarrier mediated targeting of tumor hypoxia. The first part is dedicated to the approaches by which nanocarriers could be designed to target/leverage hypoxia. These approaches include i) inhibiting Hypoxia Inducer Factor (HIF-1α); ii) hypoxia activated prodrugs/ linkers and iii) obligate anaerobe mediated targeting of tumor hypoxia. The second part, details novel nanosystems proposed to modulate tumor hypoxia through tumor oxygenation. These methods seek to lessen tumor hypoxia through vascular normalization, or reoxygenation therapy. The reoxygenation of tumor could be accomplished by: i) generation of oxygen filled nanocarriers; ii) natural/artificial oxygen nanocarriers and iii) oxygen generators. The efficacy of each approach and their potential in cancer therapy is further discussed. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-15T01:15:58.820706-05:
      DOI: 10.1002/jcp.25859
  • Mammary Alveolar Epithelial Cells Convert to Brown Adipocytes in
           Post-lactating Mice
    • Authors: Antonio Giordano; Jessica Perugini, David M. Kristensen, Loris Sartini, Andrea Frontini, Shingo Kajimura, Karsten Kristiansen, Saverio Cinti
      Abstract: During pregnancy and lactation, subcutaneous white adipocytes in the mouse mammary gland transdifferentiate reversibly to milk-secreting epithelial cells. In this study, we demonstrate by transmission electron microscopy that in the post-lactating mammary gland interscapular multilocular adipocytes found close to the mammary alveoli contain milk protein granules. Use of the Cre-loxP recombination system allowed showing that the involuting mammary gland of whey acidic protein-Cre/R26R mice, whose secretory alveolar cells express the lacZ gene during pregnancy, contains some X-Gal-stained and uncoupling protein 1–positive interscapular multilocular adipocytes. These data suggest that during mammary gland involution some milk-secreting epithelial cells in the anterior subcutaneous depot may transdifferentiate to brown adipocytes, highlighting a hitherto unappreciated feature of mouse adipose organ plasticity. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-12T23:45:31.948877-05:
      DOI: 10.1002/jcp.25858
  • Staphylococcal Protein A Promotes Osteoclastogenesis through MAPK
           Signaling During Bone Infection
    • Authors: Yuan Wang; Xin Liu, Ce Dou, Zhen Cao, Chuan Liu, Shiwu Dong, Jun Fei
      Abstract: Bone infection is a common and serious complication in the orthopedics field, which often leads to excessive bone destruction and non-union. Osteoclast is the only type of cells which have the function of bone resorption. Its over activation is closely related to excessive bone loss. Staphylococcus aureus (S.aureus) is a major pathogen causing bone infection, which can produce a large number of strong pathogenic substances staphylococcal protein A (SPA). However, few studies were reported about the effects of SPA on osteoclastogenesis. In our study, we observed that S.aureus activated osteoclasts and promoted bone loss in bone infection specimens. Then, we investigated the effects of SPA on RANKL-induced osteoclastogenesis in vitro, the results revealed that SPA promoted osteoclastic differentiation and fusion, and enhanced osteoclastic bone resorption. In addition, we also showed that SPA upregulated the expression of NFATc1 and c-FOS through the activation of MAPK signaling to promote osteoclastogenesis. Our findings might help us better understand the pathogenic role of S.aureus in bone infection and develop new therapeutic strategies for infectious bone diseases. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-10T03:15:30.811952-05:
      DOI: 10.1002/jcp.25774
  • MicroRNA-195-5p Regulates Osteogenic Differentiation of Periodontal
           Ligament Cells Under Mechanical Loading
    • Authors: Maolin Chang; Heng Lin, Haidi Fu, Beike Wang, Guangli Han, Mingwen Fan
      Abstract: Osteogenic differentiation and bone formation are tightly regulated by several factors, including microRNAs (miRNAs). However, miRNA expression patterns and function during mechanical loading-induced osteogenic differentiation of human periodontal ligament cells (PDLCs) remain unclear. Here, we investigated the differential expression of miRNA-195-5p in the periodontal tissues of mice under orthodontic mechanical loading and in primary human PDLCs exposed to a simulated tension strain. miR-195-5p was observed to be down-regulated and negatively correlated with osteogenic differentiation. Overexpression of miR-195-5p significantly inhibited PDLC differentiation under cyclic tension strain (CTS), whereas the functional inhibition of miR-195-5p yielded an opposite effect. Further experiments confirmed that WNT family member 3A (WNT3A), fibroblast growth factor 2 (FGF2) and bone morphogenetic protein receptor-1A (BMPR1A), proteins important for osteogenic activity and stability, were direct targets of miR-195-5p. Mechanical loading increased the WNT3A, FGF2 and BMPR1A protein levels, while miR-195-5p inhibited WNT3A, FGF2 and BMPR1A protein expression. WNT, FGF and BMP signaling were involved in osteogenic differentiation of PDLCs under CTS. Further study confirmed that reintroduction of WNT3A and BMPR1A can rescued the inhibition of miR-195-5p on osteogenic differentiation of PDLCs. Our findings are the first to demonstrate that miR-195-5p is a mechanosensitive gene that plays an important role in mechanical loading-induced osteogenic differentiation and bone formation. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T08:01:37.071898-05:
      DOI: 10.1002/jcp.25856
  • NSI-189, a Small Molecule with Neurogenic Properties, Exerts Behavioral
           and Neurostructural Benefits in Stroke Rats
    • Authors: Naoki Tajiri; David M. Quach, Yuji Kaneko, Stephanie Wu, David Lee, Tina Lam, Ken L. Hayama, Thomas G. Hazel, Karl Johe, Michael C. Wu, Cesar V. Borlongan
      Abstract: Enhancing neurogenesis may be a powerful stroke therapy. Here, we tested in a rat model of ischemic stroke the beneficial effects of NSI-189, an orally active, new molecular entity (mol. wt. 366) with enhanced neurogenic activity, and indicated as an anti-depressant drug in a clinical trial (Fava et al., 2015) and being tested in a Phase 2 efficacy trial (, 2016) for treatment of major depression. Oral administration of NSI-189 in adult Sprague-Dawley rats starting at 6 hours after middle cerebral artery occlusion, and daily thereafter over the next 12 weeks resulted in significant amelioration of stroke-induced motor and neurological deficits, which was maintained up to 24 weeks post-stroke. Histopathological assessment of stroke brains from NSI-189-treated animals revealed significant increments in neurite outgrowth as evidenced by MAP2 immunoreactivity that was prominently detected in the hippocampus and partially in the cortex. These results suggest NSI-189 actively stimulated remodeling of the stroke brain. Parallel in vitro studies further probed this remodeling process and demonstrated that oxygen glucose deprivation and reperfusion (OGD/R) initiated typical cell death processes, which were reversed by NSI-189 treatment characterized by significant attenuation of OGD/R-mediated hippocampal cell death and increased Ki67 and MAP2 expression, coupled with upregulation of neurogenic factors such as BDNF and SCF. These findings support the use of oral NSI-189 as a therapeutic agent well beyond the initial 6-hour time window to accelerate and enhance the overall functional improvement in the initial 6 months post stroke. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T08:00:59.936877-05:
      DOI: 10.1002/jcp.25847
  • Irisin Plays a Pivotal Role to Protect the Heart Against Ischemia and
           Reperfusion Injury
    • Authors: Hao Wang; Yu Tina Zhao, Shouyan Zhang, Patrycja M Dubielecka, Jianfeng Du, Naohiro Yano, Y. Eugene Chin, Shougang Zhuang, Gangjian Qin, Ting C Zhao
      Abstract: Irisin, a newly identified hormone, is critical to modulating body metabolism, thermogenesis and reducing oxidative stresses. However, whether irisin protects the heart against myocardial ischemia and reperfusion (I/R) injury remains unknown. In this study, we determine the effect of irisin on myocardial I/R injury in the Langendorff perfused heart and cultured myocytes. Adult C57/BL6 mice were treated with irisin (100mg/kg) or vehicle for 30 minutes to elicit preconditioning. The isolated hearts were subjected to 30 min ischemia followed by 30 min reperfusion. Left ventricular function was measured and infarction size were determined using by tetrazolium staining. Western blot was employed to determine myocardial SOD-1, active-caspase 3, annexin V, p38 and phospho-p38. H9c2 cardiomyoblasts were exposed to hypoxia and reoxygenation for assessment of the effects of irisin on mitochondrial respiration and mitochondrial permeability transition pore (mPTP). Irisin treatment produced remarkable improvements in ventricular functional recovery, as evident by the increase in RPP and attenuation in LVEDP. As compared to the vehicle treatment, irisin resulted in a marked reduction of myocardial infarct size. Notably, irisin treatment increased SOD-1 and p38 phosphorylation, but suppressed levels of active-caspase 3, cleaved PARP, and annexin V. In cardiomyoblasts exposed to hypoxia/reoxygenation, irisin treatment significantly attenuated hypoxia/reoxygenation (H/R), as indicated by the reduction of lactate dehydrogenase (LDH) leakage and apoptotic cardiomyocytes. Furthermore, irisin treatments suppressed the opening of mPTP, mitochondrial swelling, and protected mitochondria function. Our results indicate that irisin serves as a novel approach to eliciting cardioprotection, which is associated with the improvement of mitochondrial function. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T07:20:45.225788-05:
      DOI: 10.1002/jcp.25857
  • New advances of TMEM88 in cancer initiation and progression, with special
           emphasis on Wnt signaling pathway
    • Authors: Yun-xuan Ge; Chang-hui Wang, Fu-yong Hu, Lin-xin Pan, Jie Min, Kai-yuan Niu, Lei Zhang, Jun Li, Tao Xu
      Abstract: Transmembrane protein 88 (TMEM88), a newly discovered protein localized on the cell membrane. Recent studies showed that TMEM88 was involved in the regulation of several types of cancer. TMEM88 was expressed at significantly higher levels in breast cancer (BC) cell line than in normal breast cell line with co-localized with Dishevelled (DVL) in the cytoplasm of BC cell line. TMEM88 silencing in the ovarian cancer cell line CP70 resulted in significant upregulation of Wnt downstream genes (c-Myc, cyclin-D1) and other Wnt target genes including JUN, PTIX2, CTNNB1 (β-catenin), further supporting that TMEM88 inhibits canonical Wnt signaling pathway. Wnt signaling pathway has been known to play important roles in many diseases, especially in cancer. For instance, hepatocellular carcinoma (HCC) has become one of the most common tumors harboring mutations in the Wnt signaling pathway. As the inhibitor of Wnt signaling, TMEM88 has been considered to act as an oncogene or a tumor suppressor. Up-regulated TMEM88 or gene therapy approaches could be an effective therapeutic approach against tumor as TMEM88 inhibits Wnt signaling through direct interaction with DVL. Here we review the current knowledge on the functional role and potential clinical application of TMEM88 in the control of various cancers. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T00:00:40.269535-05:
      DOI: 10.1002/jcp.25853
  • Interplay between miRNAs and Human Diseases: A Review
    • Authors: Prosenjit Paul; Anindya Chakraborty, Debasree Sarkar, Malobika Langthasa, Musfhia Rahman, Minakshi Bari, RK. Sanamacha Singha, Arup Kumar Malakar, Supriyo Chakraborty
      Abstract: MicroRNAs (miRNAs) are endogenous, non-coding RNAs, which have evoked a great deal of interest due to their importance in many aspects of homeostasis and diseases. MicroRNAs are stable and are essential components of gene regulatory networks. They play a crucial role in healthy individuals and their dysregulations have also been implicated in a wide range of diseases, including diabetes, cardiovascular disease, kidney disease and cancer. This review summarized the current understanding of interactions between miRNAs and different diseases and their role in disease diagnosis and therapy. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T00:00:35.578243-05:
      DOI: 10.1002/jcp.25854
  • Adipocytes Properties and Crosstalk with Immune System in Obesity-related
    • Authors: Giulia Maurizi; Lucio Della Guardia, Angela Maurizi, Antonella Poloni
      Abstract: Obesity is a condition likely associated with several dysmetabolic conditions or worsening of cardiovascular and other chronic disturbances. A key role in this mechanism seem to be played by the onset of low-grade systemic inflammation, highlighting the importance of the interplay between adipocytes and immune system cells. Adipocytes express a complex and highly adaptive biological profile being capable to selectively activate different metabolic pathways in order to respond to environmental stimuli. It has been demonstrated how adipocytes, under appropriate stimulation, can easily differentiate and de-differentiate thereby converting themselves into different phenotypes according to metabolic necessities. Although underlying mechanisms are not fully understood, growing in adipocyte size and the inability of storing triglycerides under overfeeding conditions seem to be crucial for the switching to a dysfunctional metabolic profile, which is characterized by inflammatory and apoptotic pathways activation and by the shifting to pro-inflammatory adipokines secretion. In obesity, changes in adipokines secretion along with adipocyte deregulation and fatty acids release into circulation contribute to maintain immune cells activation as well as their infiltration into regulatory organs. Over the well-established role of macrophages, recent findings suggest the involvement of new classes of immune cells such as T regulatory lymphocytes and neutrophils in the development inflammation and multi systemic worsening. Deeply understanding the pathways of adipocyte regulation and the de-differentiation process could be extremely useful for developing novel strategies aimed at curbing obesity-related inflammation and related metabolic disorders. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-09T00:00:31.24522-05:0
      DOI: 10.1002/jcp.25855
  • Emerging Roles of ER Stress and Unfolded Protein Response Pathways in
           Skeletal Muscle Health and Disease
    • Authors: Kyle R. Bohnert; Joseph D. McMillan, Ashok Kumar
      Abstract: Skeletal muscle is the most abundant tissue in the human body and can adapt its mass as a consequence of physical activity, metabolism, growth factors, and disease conditions. Skeletal muscle contains an extensive network of endoplasmic reticulum (ER), called sarcoplasmic reticulum, which plays an important role in the regulation of proteostasis and calcium homeostasis. In many cell types, environmental and genetic factors that disrupt ER function cause an accumulation of misfolded and unfolded proteins in the ER lumen that ultimately leads to ER stress. To alleviate the stress and restore homeostasis, the ER activates a signaling network called the unfolded protein response (UPR). The UPR has three arms, which regulate protein synthesis and expression of many ER chaperone and regulatory proteins. However, the role of individual UPR pathways in skeletal muscle has just begun to be investigated. Recent studies suggest that UPR pathways play pivotal roles in muscle stem cell homeostasis, myogenic differentiation, and regeneration of injured skeletal muscle. Moreover, markers of ER stress and the UPR are activated in skeletal muscle in diverse conditions such as exercise, denervation, starvation, high fat diet, cancer cachexia, and aging. Accumulating evidence also suggests that ER stress may have important roles in the pathogenesis of inflammatory myopathies and genetic muscle disorders. The purpose of this review article is to discuss the role and potential mechanisms by which ER stress and the individual arms of the UPR regulate skeletal muscle formation, plasticity, and function in various physiological and pathophysiological conditions. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-08T06:35:32.518742-05:
      DOI: 10.1002/jcp.25852
  • Role of Parp Activity in Lung Cancer-induced Cachexia: Effects on Muscle
           Oxidative Stress, Proteolysis, Anabolic Markers and Phenotype
    • Authors: Alba Chacon-Cabrera; Mercè Mateu-Jimenez, Klaus Langohr, Clara Fermoselle, Elena García-Arumí, Antoni L. Andreu, Jose Yelamos, Esther Barreiro
      Abstract: Strategies to treat cachexia are still at its infancy. Enhanced muscle protein breakdown and ubiquitin-proteasome system are common features of cachexia associated with chronic conditions including lung cancer (LC). Poly(ADP-ribose) polymerases (PARP), which play a major role in chromatin structure regulation, also underlie maintenance of muscle metabolism and body composition. We hypothesized that protein catabolism, proteolytic markers, muscle fiber phenotype, and muscle anabolism may improve in respiratory and limb muscles of LC-cachectic Parp-1-deficient (Parp-1-/-) and Parp-2-/- mice. In diaphragm and gastrocnemius of LC (LP07 adenocarcinoma) bearing mice (wild type, Parp-1-/- and Parp-2-/-), PARP activity (ADP-ribose polymers, pADPr), redox balance, muscle fiber phenotype, apoptotic nuclei, tyrosine release, protein ubiquitination, muscle-specific E3 ligases, NFkB signaling pathway, markers of muscle anabolism (Akt, mTOR, p70S6K, and mitochondrial DNA) were evaluated along with body and muscle weights and limb muscle force. Compared to wild type cachectic animals, in both respiratory and limb muscles of Parp-1-/- and Parp-2-/- cachectic mice: cancer induced-muscle wasting characterized by increased PARP activity, protein oxidation, tyrosine release, and ubiquitin-proteasome system (total protein ubiquitination, atrogin-1, and 20S proteasome C8 subunit) were blunted, the reduction in contractile myosin and atrophy of the fibers was attenuated, while no effects were seen in other structural features (inflammatory cells, internal or apoptotic nuclei), and markers of muscle anabolism partly improved. Activation of either PARP-1 or -2 is likely to play a role in muscle protein catabolism via oxidative stress, NF-kB signaling, and enhanced proteasomal degradation in cancer-induced cachexia. Therapeutic potential of PARP activity inhibition deserves attention. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-08T06:31:05.604909-05:
      DOI: 10.1002/jcp.25851
  • The SGK1 Inhibitor SI113 Induces Autophagy, Apoptosis and Endoplasmic
           Reticulum Stress in Endometrial Cancer Cells
    • Authors: Domenico Conza; Paola Mirra, Gaetano Calì, Teresa Tortora, Luigi Insabato, Francesca Fiory, Silvia Schenone, Rosario Amato, Francesco Beguinot, Nicola Perrotti, Luca Ulianich
      Abstract: Endometrial cancer is often characterized by PI3K/AKT pathway deregulation. Recently it has been suggested that SGK1, a serine/threonine protein kinase that shares structural and functional similarities with the AKT family, might play a role in cancer, since its expression and/or activity has been found to be deregulated in different human tumors. However, the role of SGK1 in endometrial cancer has been poorly investigated. Here we show that SGK1 expression is increased in tissue specimens from neoplastic endometrium. The SGK1 inhibitor SI113 induced a significant reduction of endometrial cancer cells viability, measured by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. This effect was associated to the increase of autophagy, as revealed by the increase of the markers LC3B-II and beclin I, detected by both immunofluorescence and western blot analysis. SI113 treatment caused also apoptosis of endometrial cancer cells, evidenced by the cleavage of the apoptotic markers PARP and Caspase-9. Intriguingly, these effects were associated to the induction of endoplasmic reticulum stress markers GRP78 and CHOP evaluated by both Real-Time RT-PCR and Western Blot analysis. Increased expression of SGK1 in endometrial cancer tissues suggest a role for SGK1 in this type of cancer, as reported for other malignancies. Moreover, the efficacy of SI113 in affecting endometrial cancer cells viability, possibly via endoplasmic reticulum stress activation, identifies SGK1 as an attractive molecular target for new tailored therapeutic intervention for the treatment of endometrial cancer. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-08T06:20:39.434719-05:
      DOI: 10.1002/jcp.25850
  • Activating Transcription Factor 3 (ATF3) Protects Against
           Lipopolysaccharide-Induced Acute Lung Injury Via Inhibiting the Expression
           of TL1A
    • Authors: Lanlan Qian; Yunfeng zhao, Liang Guo, Shaoying Li, Xueling Wu
      Abstract: Excessive inflammatory responses are critical in the pathogenesis of acute lung injury (ALI). Activating transcription factor 3 (ATF3) is a stress-induced transcriptional regulator that is a negative regulator of inflammatory responses. Therefore, we investigated the role and signaling pathways of ATF3 in lipopolysaccharide (LPS)-induced ALI in mice. The mouse macrophage RAW264.7 cells were cultured on HTS 24-Transwell filter plates in presence of ATF3 siRNA before exposure to LPS. ATF3 knock-out (KO) and wild type (WT) mice were challenged by intra-peritoneal injection of LPS (15mg/kg). Gene analysis was used to analyze differential gene expression between ATF3 KO and WT mice. LPS increased the expression of ATF3 in RAW264.7 cells and in lung tissues of mice, The concentration of TNFα and IL-6 was significantly increased in ATF3 siRNA-treated RAW264.7 cells compared to control cells after LPS stimulation. The concentration of TNFα, IL-6 and IL-1β in serum and lung tissue of ATF3 KO mice was significantly increased compared to ATF3 WT mice. In addition, the lung wet/dry weight and BALF protein were significantly increased in ATF3 KO mice after LPS injection at 6, 24 and 48-hours. The survival of ATF3 KO mice significantly decreased. Differential gene analysis showed that TL1A was highly expressed in LPS-induced lung tissues of ATF3 KO mice.Moreover, ATF3 down-regulated the expression of TL1A in RAW264.7 cells and in lung tissues. These findings suggest that ATF3 protects against LPS-induced ALI via inhibiting TL1A expression. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-08T06:20:36.958384-05:
      DOI: 10.1002/jcp.25849
  • Downregulation of Methyltransferase Dnmt2 Results in Condition-dependent
           Telomere Shortening and Senescence or Apoptosis in Mouse Fibroblasts
    • Authors: Anna Lewinska; Jagoda Adamczyk-Grochala, Ewa Kwasniewicz, Maciej Wnuk
      Abstract: Dnmt2 is a highly conserved methyltransferase of uncertain biological function(s). As Dnmt2 was considered as a driver of fruit fly longevity and a modulator of stress response, we decided to evaluate the role of Dnmt2 during stress-induced premature senescence in NIH3T3 mouse fibroblasts. Stable knockdown of Dnmt2 resulted in hydrogen peroxide-mediated sensitivity and apoptosis, whereas in the control conditions, senescence was induced. Cellular senescence was accompanied by elevated levels of p53 and p21, decreased telomere length and telomerase activity, increased production of reactive oxygen species and protein carbonylation, and DNA damage. Dnmt2 silencing also promoted global DNA and RNA hypermethylation, and upregulation of methyltransferases, namely Dnmt1, Dnmt3a and Dnmt3b. Taken together, we show for the first time that Dnmt2 may promote lifespan in the control conditions and survival during stress conditions in mouse fibroblasts. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-08T06:20:30.164815-05:
      DOI: 10.1002/jcp.25848
  • Sec16 in Conventional and Unconventional Exocytosis: Working at the
           Interface of Membrane Traffic and Secretory Autophagy'
    • Authors: Bor Luen Tang
      Abstract: Sec16 is classically perceived to be a scaffolding protein localized to the transitional endoplasmic reticulum (tER) or the ER exit sites (ERES), and has a conserved function in facilitating coat protein II (COPII) complex-mediated ER exit. Recent findings have however pointed towards a role for Sec16 in unconventional exocytosis of certain membrane proteins, such as the Cystic fibrosis transmembrane conductance regulator (CFTR) in mammalian cells, and possibly also α-integrin in certain contexts of Drosophila development. In this regard, Sec16 interacts with components of a recently deciphered pathway of stress-induced unconventional exocytosis, which is dependent on the tether protein Golgi reassembly stacking proteins (GRASPs) and the autophagy pathway. Intriguingly, Sec16 also appears to be post-translationally modified by autophagy-related signaling processes. Sec16 is known to be phosphorylated by the atypical Extracellular signal regulated kinase 7 (Erk7) upon serum and amino acid starvation, both represent conditions that trigger autophagy. Recent work has also shown that Sec16 is phosphorylated, and thus regulated by the prominent autophagy-initiating Unc-51-like autophagy activating kinase 1 (Ulk1), as well as another autophagy modulator Leucine-rich repeat kinase 2 (Lrrk2). The picture emerging from Sec16's network of physical and functional interactors allows the speculation that Sec16 is situated (and may in yet undefined ways function) at the interface between COPII-mediated exocytosis of conventional vesicular traffic and the GRASP/autophagy-dependent mode of unconventional exocytosis. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-04T06:30:58.704378-05:
      DOI: 10.1002/jcp.25842
  • Molecular Pathogenesis in Chronic Obstructive Pulmonary Disease and
           Therapeutic Potential by Targeting AMP-activated Protein Kinase
    • Authors: Zhihui Zhang; Xiaoyu Cheng, Li Yue, Wenhui Cui, Wencheng Zhou, Jian Gao, Hongwei Yao
      Abstract: Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide, which is characterized by a persistent airflow limitation caused by chronic inflammatory responses to noxious particles or gases. Cigarette smoke and environmental pollutions are major etiological factors for causing COPD. It has been shown that cigarette smoking causes abnormal inflammatory responses, cellular senescence, mitochondrial dysfunction and metabolic dysregulation, suggesting their involvement in the development of COPD. Although the medical care and treatment have advanced, there are no effective therapies to stop or reverse lung destruction in COPD/emphysema. AMP-activated protein kinase (AMPK) is a serine threonine kinase with α, β, and γ subunits that are highly conserved through evolution. AMPK has been shown to regulate bioenergetics, inflammatory responses, senescence, and metabolism. This review focused on the updated understanding of molecular pathogenesis of COPD, and highlighted the crucial roles of AMPK in lung abnormalities as well as discussed the potential therapeutics of AMPK activators in preventing and halting the progression of COPD. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-04T06:30:57.265612-05:
      DOI: 10.1002/jcp.25844
  • NMDA Receptor Subunit Composition Controls Dendritogenesis of Hippocampal
           Neurons through CAMKII, CREB-P and H3K27ac
    • Authors: Fernando J. Bustos; Nur Jury, Pablo Martinez, Estibaliz Ampuero, Matias Campos, Sebastian Abarzúa, Karen Jaramillo, Susanne Ibing, Muriel D. Mardones, Henny Haensgen, Julia Kzhyshkowska, Maria Florencia Tevy, Rachael Neve, Magdalena Sanhueza, Lorena Varela-Nallar, Martín Montecino, Brigitte van Zundert
      Abstract: Dendrite arbor growth, or dendritogenesis, is choreographed by a diverse set of cues, including the NMDA receptor (NMDAR) subunits NR2A and NR2B. While NR1NR2B receptors are predominantly expressed in immature neurons and promote plasticity, NR1NR2A receptors are mainly expressed in mature neurons and induce circuit stability. How the different subunits regulate these processes is unclear, but this is likely related to the presence of their distinct C-terminal sequences that couple different signaling proteins. Calcium-calmodulin-dependent protein kinase II (CaMKII) is an interesting candidate as this protein can be activated by calcium influx through NMDARs. CaMKII triggers a series of biochemical signaling cascades, involving the phosphorylation of diverse targets. Among them, the activation of cAMP response element-binding protein (CREB-P) pathway triggers a plasticity-specific transcriptional program through unknown epigenetic mechanisms. Here we found that dendritogenesis in hippocampal neurons is impaired by several well-characterized constructs (i.e. NR2B-RS/QD) and peptides (i.e. tatCN21) that specifically interfere with the recruitment and interaction of CaMKII with the NR2B C-terminal domain. Interestingly, we found that transduction of NR2AΔIN, a mutant NR2A construct with increased interaction to CaMKII, reactivates dendritogenesis in mature hippocampal neurons in vitro and in vivo. To gain insights into the signaling and epigenetic mechanisms underlying NMDAR-mediated dendritogenesis, we used immunofluorescence staining to detect CREB-P and acetylated lysine 27 of histone H3 (H3K27ac), an activation-associated histone tail mark. In contrast to control mature neurons, our data shows that activation of the NMDAR/CaMKII/ERK-P/CREB-P signaling axis in neurons expressing NR2AΔIN is not correlated with increased nuclear H3K27ac levels. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-04T06:30:55.376317-05:
      DOI: 10.1002/jcp.25843
  • A Prognostic 4-Gene Expression Signature for Squamous Cell Lung Carcinoma
    • Authors: Jun Li; Jing Wang, Yanbin Chen, Lijie Yang, Sheng Chen
      Abstract: Squamous cell lung carcinoma (SQCLC), a common and fatal subtype of lung cancer, caused lots of mortalities and showed different outcomes in prognosis. This study was to screen key genes and to figure a prognostic signature to cluster the patients with SQCLC. RNA-Seq data from 550 patients with SQCLC were downloaded from The Cancer Genome Atlas (TCGA). Genetically changed genes were identified and analyzed in univariate survival analysis. Genes significantly influencing prognosis were selected with frequency higher than 100 in lasso regression. Meanwhile, area under the curve (AUC) values and hazard ratios (HR) for seed genes were obtained with R Language. Functional enrichment analysis was performed and clustering effectiveness of the selected common gene set was analyzed with Kaplan-meier. Finally, the stability and validity of the optimal clustering model were verified. A total of 7222 genetically changed genes were screened, including 1045 ones with p 
      PubDate: 2017-02-04T06:30:53.303521-05:
      DOI: 10.1002/jcp.25846
  • IL-10 Expression in Macrophages from Neonates Born from Obese Mothers Is
           Suppressed by IL-4 and LPS/INFγ
    • Authors: Francisca Cifuentes-Zúñiga; Viviana Arroyo-Jousse, Gustavo Soto-Carrasco, Paola Casanello, Ricardo Uauy, Bernardo J. Krause, José A. Castro-Rodríguez
      Abstract: Obese women offspring have a higher risk of developing chronic diseases associated with an altered immune function. We aim to determine, in neonatal monocyte-derived macrophages, whether maternal obesity is associated with an altered expression and DNA methylation of pro- and anti-inflammatory genes, along with a higher pro-inflammatory response. Cord blood from newborns of obese (Ob) and lean (control) women were obtained at delivery. Monocytes were isolated and differentiated into macrophages, in which M1 (LPS/IFNγ) and M2 (IL-4) polarization were assayed. The mRNA levels for TNFα, IL-1β, IL-12A, IL-12B, IL-10 and IL-4R were quantified by qPCR and the DNA methylation of candidate genes determined by pyrosequencing. Results: Ob-monocytes had decreased levels of mRNA for pro-inflammatory cytokines IL-1β, IL-10 and IL-12B compared with controls. Conversely, Ob-macrophages showed increased levels of mRNA for TNFα, IL-4R and IL-10 compared with controls. M1 response was comparable between both groups, characterized by an important induction of TNFα and IL-1β. In response to an M2 stimulus, control macrophages showed a decreased expression of inflammatory mediators whilst Ob-macrophages had an additional suppression of the anti-inflammatory mediator IL-10. Changes in IL-1β (monocytes) and IL-10 (macrophages) in Ob-monocytes were paralleled by changes in their promoter DNA methylation in fetal monocytes. These results suggest that monocyte-derived macrophages from obese newborns show a basal anti-inflammatory phenotype with an unbalanced response to M1 and M2 polarization stimuli. The presence of changes in DNA methylation of key inflammatory genes in neonatal monocytes suggests an intrauterine programing of immune function by maternal obesity. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-04T06:30:45.23629-05:0
      DOI: 10.1002/jcp.25845
  • Deubiquitinating enzyme USP22 positively regulates c-Myc stability and
           tumorigenic activity in mammalian and breast cancer cells
    • Authors: Dongyeon Kim; Ahyoung Hong, Hye In Park, Woo Hyun Shin, Lang Yoo, Seo Jeong Jeon, Kwang Chul Chung
      Abstract: The proto-oncogene c-Myc has a pivotal function in growth control, differentiation and apoptosis and is frequently affected in human cancer, including breast cancer. Ubiquitin-specific protease 22 (USP22), a member of the USP family of deubiquitinating enzymes (DUBs), mediates deubiquitination of target proteins, including histone H2B and H2A, telomeric repeat binding factor 1, and cyclin B1. USP22 is also a component of the mammalian SAGA transcriptional co-activating complex. In this study, we explored the functional role of USP22 in modulating c-Myc stability and its physiological relevance in breast cancer progression. We found that USP22 promotes deubiquitination of c-Myc in several breast cancer cell lines, resulting in increased levels of c-Myc. Consistent with this, USP22 knockdown reduces c-Myc levels. Furthermore, overexpression of USP22 stimulates breast cancer cell growth and colony formation, and increases c-Myc tumorigenic activity. In conclusion, the present study reveals that USP22 in breast cancer cell lines increases c-Myc stability through c-Myc deubiquitination, which is closely correlated with breast cancer progression. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-04T06:30:35.408356-05:
      DOI: 10.1002/jcp.25841
  • Involvement of CX3CL1 in the Migration of Osteoclast Precursors Across
           Osteoblast Layer Stimulated by Interleukin-1ß
    • Authors: Tsuyoshi Matsuura; Shizuko Ichinose, Masako Akiyama, Yuki Kasahara, Noriko Tachikawa, Ken-ichi Nakahama
      Abstract: The trigger for bone remodeling is bone resorption by osteoclasts. Osteoclast differentiation only occurs on the old bone, which needs to be repaired under physiological conditions. However, uncontrolled bone resorption is often observed in pro-inflammatory bone diseases, such as rheumatoid arthritis. Mature osteoclasts are multinuclear cells that differentiate from monocyte/macrophage lineage cells by cell fusion. Although Osteoclast precursors should migrate across osteoblast layer to reach bone matrix before maturation, the underlying mechanisms have not yet been elucidated in detail. We herein found that osteoclast precursors utilize two routes to migrate across osteoblast layer by confocal- and electro-microscopic observations. The osteoclast supporting activity of osteoblasts inversely correlated with osteoblast density and was positively related to the number of osteoclast precursors under the osteoblast layer. Osteoclast differentiation was induced by IL-1ß, but not by PGE2 in high-density osteoblasts. Osteoblasts and osteoclast precursors expressed CX3CL1 and CX3CR1, respectively, and the expression of CX3CL1 increased in response to interleukin-1ß. An anti-CX3CL1-neutralizing antibody inhibited the migration of osteoclast precursors and osteoclast differentiation. These results strongly suggest the involvement of CX3CL1 in the migration of osteoclast precursors and osteoclastogenesis, and will contribute to the development of new therapies for bone diseases. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc.Transcellular and paracellular migration of osteoclast precursors across osteoblast layer were observed. CX3CL1 was involved in the migration of osteoclast precursors and osteoclastogenesis.
      PubDate: 2017-02-03T10:00:30.140846-05:
      DOI: 10.1002/jcp.25577
  • Effect of Stress on Human Biology: Epigenetics, Adaptation, Inheritance
           and Social Significance
    • Authors: David Denhardt
      Abstract: I present a brief introduction to epigenetics, focused primarily on methylation of the genome and various regulatory RNAs, modifications of associated histones, and their importance in enabling us to adapt to real and changing environmental, developmental and social circumstances. Following this is a more extensive overview of how it impacts our inheritance, our entire life (which changes as we age), and how we interact with others. Throughout I emphasize the critical influence that stress, of many varieties exerts, via epigenetic means, on much of how we live and survive, mostly in the brain. I end with a short section on multigenerational transmission, drugs, and the importance of both social life and early life experiences in the development of adult diseases. There will be nothing about cancer. Though epigenetics is critical in that field, it is a whole different cobweb of complications (some involving stress). This article is protected by copyright. All rights reserved
      PubDate: 2017-02-03T05:25:39.058122-05:
      DOI: 10.1002/jcp.25837
  • 4PBA Strongly Attenuates Endoplasmic Reticulum Stress, Fibrosis and
           Mitochondrial Apoptosis Markers in Cyclosporine Treated Human Gingival
    • Authors: Suresh Ranga Rao; Subbarayan Rajasekaran, Supraja Ajitkumar, Murugan Girija Dinesh
      Abstract: Cyclosporine induces overgrowth of human gingiva. Previously we have shown (i) cyclosporine - inducing ER stress in human gingival fibroblasts, (ii) increased matrix protein expression, and (iii) interference with mitochondrial pro- and anti - apoptotic factors. This study was undertaken to assess the effects of melatonin (an antioxidant), 4PBA (an ER stress inhibitor), and simvastatin on the expression of ER Stress markers, as well as on matrix and mitochondrial markers. Human gingival fibroblasts incubated with cyclosporine, or without melatonin/4PBA/statin. After 24 hours of incubation, mRNA expression of ER stress markers (GRP78, CHOP, XBP1 and XBPs) and, matrix protein markers (like α-SMA, VEGF, TGF-β, CTGF) and mitochondrial apoptosis markers estimated and compared with housekeeping gene GAPDH. Compared to the control cyclosporine significantly augmented ER Stress and matrix proteins, which decreased significantly with the use of melatonin,4PBA, and simvastatin. The mitochondrial proapoptotic molecule cyclophilin D, as well as Bcl2 expression, also decreased after PBA treatment, paralleling an increase in cytochrome c expression. The effect of 4PBA was much more pronounced than the influence of other two. In conclusion, 4PBA could be a viable therapeutic option for drug-induced gingival overgrowth. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-03T05:20:52.740403-05:
      DOI: 10.1002/jcp.25836
  • Mimicking Exercise in Three-Dimensional Bioengineered Skeletal Muscle to
    • Authors: Andreas M. Kasper; Daniel C. Turner, Neil R.W. Martin, Adam P. Sharples
      Abstract: Bioengineering of skeletal muscle in-vitro in order to produce highly aligned myofibres in relevant three dimensional (3D) matrices have allowed scientists to model the in-vivo skeletal muscle niche. This review discusses essential experimental considerations for developing bioengineered muscle in order to investigate exercise mimicking stimuli. We identify current knowledge in the use of electrical stimulation and co-culture with motor neurons to enhance skeletal muscle maturation and contractile function in bioengineered systems in-vitro. Importantly, we provide a current opinion on the use of acute and chronic exercise mimicking stimuli (electrical stimulation and mechanical overload) and the subsequent mechanisms underlying physiological adaptation in 3D bioengineered muscle. We also identify that future studies using the latest bioreactor technology, providing simultaneous electrical and mechanical loading and flow perfusion in-vitro, may provide the basis for advancing knowledge in the future. We also envisage, that more studies using genetic, pharmacological and hormonal modifications applied in human 3D bioengineered skeletal muscle may allow for an enhanced discovery of the in-depth mechanisms underlying the response to exercise in relevant human testing systems. Finally, 3D bioengineered skeletal muscle may provide an opportunity to be used as a pre-clinical in-vitro test-bed to investigate the mechanisms underlying catabolic disease, whilst modelling disease itself via the use of cells derived from human patients without exposing animals or humans (in phase I trials) to the side effects of potential therapies. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-03T05:20:38.172054-05:
      DOI: 10.1002/jcp.25840
  • Reduced Primary Cilia Length and Altered Arl13b Expression Are Associated
           with Deregulated Chondrocyte Hedgehog Signalling in Alkaptonuria
    • Authors: Stephen D Thorpe; Silvia Gambassi, Clare L Thompson, Charmilie Chandrakumar, Annalisa Santucci, Martin M Knight
      Abstract: Alkaptonuria (AKU) is a rare inherited disease resulting from a deficiency of the enzyme homogentisate 1,2-dioxygenase which leads to the accumulation of homogentisic acid (HGA). AKU is characterised by severe cartilage degeneration, similar to that observed in osteoarthritis. Previous studies suggest that AKU is associated with alterations in cytoskeletal organisation which could modulate primary cilia structure/function. This study investigated whether AKU is associated with changes in chondrocyte primary cilia and associated Hedgehog signalling which mediates cartilage degradation in osteoarthritis. Human articular chondrocytes were obtained from healthy and AKU donors. Additionally, healthy chondrocytes were treated with HGA to replicate AKU pathology (+HGA). Diseased cells exhibited shorter cilia with length reductions of 36% and 16% in AKU and +HGA chondrocytes respectively when compared to healthy controls. Both AKU and +HGA chondrocytes demonstrated disruption of the usual cilia length regulation by actin contractility. Furthermore, the proportion of cilia with axoneme breaks and bulbous tips was increased in AKU chondrocytes consistent with defective regulation of ciliary trafficking. Distribution of the hedgehog-related protein Arl13b along the ciliary axoneme was altered such that its localisation was increased at the distal tip in AKU and +HGA chondrocytes. These changes in cilia structure/trafficking in AKU and +HGA chondrocytes were associated with a complete inability to activate hedgehog signalling in response to exogenous ligand. Thus, we suggest that altered responsiveness to Hedgehog, as a consequence of cilia dysfunction, may be a contributing factor in the development of arthropathy highlighting the cilium as a novel target in AKU. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-03T05:20:34.924078-05:
      DOI: 10.1002/jcp.25839
  • Metformin Attenuates Albumin-induced Alterations in Renal Tubular Cells In
    • Authors: Soumaya Allouch; Shankar Munusamy
      Abstract: Proteinuria (albuminuria) plays a crucial role in the etiology of chronic kidney disease (CKD) via alteration of multiple signaling pathways and cellular process in renal cells. The objectives of this study are to investigate the effects of activation of the energy-sensing molecule AMP-activated kinase (AMPK) in renal cells using metformin on endoplasmic reticulum (ER) stress, AKT, mTOR, epithelial-to-mesenchymal transition (EMT), autophagy and apoptosis that are thought to mediate renal cell injury during proteinuria, and to dissect the AMPK- and non-AMPK mediated effects of metformin using an in vitro model of albumin-induced renal cell injury. Rat renal proximal tubular (NRK-52E) cells were exposed to 10 and 15 mg/ml of albumin for 72 hours in the presence of 1 mM Metformin and/or 0.5 µM compound C, and assessed for alterations in the aforementioned pathways. Metformin treatment restored AMPK phosphorylation and augmented autophagy in renal cells exposed to albumin. In addition, metformin treatment attenuated the albumin-induced phosphorylation of AKT and the downstream targets of mTOR and prevented albumin-mediated inductions of EMT marker (α-SMA), pro-apoptotic ER stress marker CHOP and apoptotic caspases -12 and -3 in renal cells. Blockade of metformin-induced AMPK activation with compound C blunted the ER defense response and autophagy but had no effect on the markers of EMT and apoptosis in our model. Our studies suggest that metformin protects renal cells against proteinuric cytotoxicity via suppression of AKT and mTOR activation, inhibition of EMT and apoptosis, and augmentation of autophagy and ER defense response through AMPK-independent and AMPK-dependent mechanisms respectively. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-03T05:20:33.86013-05:0
      DOI: 10.1002/jcp.25838
  • Osteogenic Differentiation of Human Gingival Mesenchymal Stem Cells by
           Aristolochia bracteolata Supplementation through Enhanced Runx2 Expression
    • Authors: Dinesh Murugan Girija; Suresh Ranga Rao, Mangathayaru Kalachaveedu, Rajasekaran Subbarayan
      Abstract: The phenotypic characteristics of human gingival derived mesenchymal stem cells (HGMSCs) on induction with total methanol extract of Aristolochia bracteolata have been evaluated. HGMSCs were cultured in control and two different induction medium: Control medium (basal medium), OM1 (Standard induction medium), and OM2 (100µg/ml of Aristolochia bracteolata). Osteogenic differentiation of the cultured cells was assessed by studying the calcium deposition and osteoblastic gene expression. OM2 medium showed an enhanced osteogenic differentiation potential than OM1 as measured by increased calcium deposition and elevated expression of Runx2, Osteopontin, osteonectin, osteocalcin, Collagen type I and ALP levels in comparison with OM1 differentiated cells. We conclude that at 100µg/ml A.bracteolata has induced HGMSC differentiation into osteogenic lineage consequent to enhanced Runx2 expression and related osteogenic genes. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-02T06:16:40.551568-05:
      DOI: 10.1002/jcp.25835
  • Brain-Derived Neurotrophic Factor Improves Proliferation of Endometrial
           Epithelial Cells by Inhibition of Endoplasmic Reticulum Stress During
           Early Pregnancy
    • Authors: Whasun Lim; Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song
      Abstract: Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family binds to two transmembrane receptors; neurotrophic receptor tyrosine kinase 2 (NTRK2) with high affinity and p75 with low affinity. Although BDNF-NTRK2 signaling in the central nervous system is known, signaling in the female reproductive system is unknown. Therefore, we determined effects of BDNF on porcine endometrial luminal epithelial (pLE) cells isolated from Day 12 of pregnancy, as well as expression of BDNF and NTRK2 in endometria of cyclic and pregnant pigs. BDNF-NTRK2 genes were expressed in uterine glandular (GE) and luminal (LE) epithelia during early pregnancy. In addition, their expression in uterine GE and LE decreased with increasing parity of sows. Recombinant BDNF increased proliferation in pLE cells in a dose-dependent, as well as expression of PCNA and Cyclin D1 in nuclei of pLE cells. BDNF also activated phosphorylation of AKT, P70S6K, S6, ERK1/2, JNK, P38 proteins in pLE cells. In addition, cell death resulting from tunicamycin-induced ER stress was prevented when pLE cells were treated with the combination of tunicamycin and BDNF which also decreased cells in the Sub-G1 phase of the cell cycle. Furthermore, tunicamycin-induced unfolded protein response genes were mostly down-regulated to the basal levels as compared to non-treated pLE cells. Our finding suggests that BDNF acts via NTRK2 to induce development of pLE cells for maintenance of implantation and pregnancy by activating cell signaling via the PI3K and MAPK pathways and by inhibiting ER stress. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-02T06:16:06.664023-05:
      DOI: 10.1002/jcp.25834
  • sp2-Iminosugar α-Glucosidase Inhibitor
           1-C-octyl-2-oxa-3-oxocastanospermine Specifically Affected Breast Cancer
           Cell Migration through Stim1, β1-Integrin, and FAK Signaling Pathways
    • Authors: Nahla Gueder; Ghada Allan, Marie-Sophie Telliez, Frédéric Hague, José M Garcia Fernandez, Elena M. Sanchez-Fernandez, Carmen Ortiz-Mellet, Ahmed Ahidouch, Halima Ouadid-Ahidouch
      Abstract: Aberrant glycosylation changes on many glycoproteins are often related to cancer progression and metastasis. sp2-Iminosugar-type castanospermine analogues, inhibitors of α-glucosidases, have been reported to exhibit antitumor activity. However, their effects on cell migration and the underlying molecular mechanism are not fully understood. Here, we investigated the effect of the pseudo- C-octyl glycoside 2-oxa-3-oxocastanospermine derivatives (CO-OCS) on breast cancer cells (MCF-7 and MDA-MB-231 cells), and MCF-10A mammary normal cell lines. We showed that CO-OCS treatment results in the drastic decrease of breast cancer cell migration without affecting cell proliferation. Furthermore, CO-OCS significantly reduced both the expression of β1-integrin, which is a crucial interacting partner of Focal Adhesion Kinase (FAK), and the phosphorylation rates of FAK and ERK1/2. CO-OCS also drastically reduced Ca2+ entry through Store Operated Channels (SOC). Orai1 and Stim1, two N-glycosylated proteins, are involved in Store-Operated Calcium Entry (SOCE), and are essential for breast tumor cell migration. Our results showed that CO-OCS decreased the expression, at the protein level, of Stim1 without affecting that of Orai1. Moreover, cell migration and SOCE were attenuated by CO-OCS as well as when Stim1 was silenced. In contrast, in MCF-10A cells, CO-OCS slightly reduced cell migration, but was without effect on gene expression of Stim1, Orai1, β1-integrin or FAK and ERK1/2 activation. Our results provide strong evidence for a significant effect of CO-OCS on breast cancer cell migration and support that this effect was associated with β1-integrin, Stim1 and FAK signaling pathways. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-01T07:07:49.931355-05:
      DOI: 10.1002/jcp.25832
  • GD2-Targeted Immunotherapy and Potential Value of Circulating microRNAs in
    • Authors: Sharareh Gholamin; Hamed Miezaei, Seyed-Mostafa Razavi, Seyed Mahdi Hassanian, Leila Saadatpour, Aria Masoudifar, Soodabeh ShahidSales, Amir Avan
      Abstract: Neuroblastoma (NB) with various clinical presentation is a known childhood malignancy. Despite significant progress in treatment of NB afflicted patients, high risk disease is usually associated with poor outcome, resulting in long-term survival of less that 50%. Known as a disease most commonly originated form the nerve roots, the variants involved in NB imitation and progression remain to be elucidated. The outcome of low to intermediate risk disease is favorable whereas the high risk NB disease with dismal prognosis, positing the necessity of novel approaches for early detection and prognostication of advanced disease. Tailored immunotherapy approaches have shown significant improvement in high-risk NB patients. It has found a link between Gangliosides and progression of NB. The vast majority of neuroblastoma tumors express elevated levels of GD2, opening new insight into using anti-GD2 drugs as potential treatments for NBs. Implication of anti-GD2 monoclonal antibodies for treatment of high risk NBs triggers further investigation to unearth novel biomarkers as prognostic and response biomarker to guide additional multimodal tailored treatment approaches. A growing body of evidence supports the usefulness of miRNAs to evaluate high risk NBs response to anti-GD2 drugs and further prevent drug-related toxicities in refractory or recurrent NBs. miRNAs and circulating proteins in body fluids (plasma, and serum) present as potential biomarkers in early detection of NBs. Here, we summarize various biomarkers involved in diagnosis, prognosis and response to treatment in patients with NB. We further attempted to overview prognostic biomarkers in response to treatment with anti-GD2 drugs. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-01T07:07:39.738355-05:
      DOI: 10.1002/jcp.25793
  • Methylated of Genes Behaving as Potential Biomarkers in Evaluating
           Malignant Degree of Glioblastoma
    • Authors: Wan-Shun Wen; Sheng-Li Hu, Zhibing Ai, Lin Mou, Jing-Min Lu, Sen Li
      Abstract: Abnormal methylation genes usually act as oncogenes or anti-oncogenes in the occurrence and development of tumor, indicating their potential role as biomarkers in the evaluation of malignant tumor. However, the research on methylation's association with glioblastoma was rare. We attempted to figure out whether the methylation of genes could serve as the biomarker in evaluating the malignant degree of GBM. Methylation microarray data of 275 GBM patients have been downloaded from The Cancer Genome Atlas (TCGA) dataset. Logistic regression was used to find the methylated genes associated with the malignant degree of patients with the tumor. Functional enrichment analysis and network analysis were further performed on these selected genes.A total of 668, 412, 470 and 620 genes relevant with the methylation or demethylation were found to be associated with the malignant degree, Grade1,Grade2,Grade3,Grade4 of tumor. The higher the degree of malignant tumor, the higher of its methylation degree of its corresponding genes. GO and KEGG analysis results showed that these methylated genes were enriched in many functions as cell adhesion, abnormal transcription and cell cycle disorder, etc. Of note, CCL11 and LCN11 were found to be significantly related to the progression of GBM.Critical genes associated with cell cycle as CCL11 and LCN1 may play essential roles in the occurrence, development, and transition of glioblastoma. More research was needed to explore its potential molecular mechanism. This article is protected by copyright. All rights reserved
      PubDate: 2017-02-01T07:07:29.521057-05:
      DOI: 10.1002/jcp.25831
  • DNA Methylation Profiling in Chondrocyte Dedifferentiation In Vitro
    • Authors: Li Duan; Yujie Liang, Bin Ma, Daming Wang, Wei Liu, Jianghong Huang, Jianyi Xiong, Liangquan Peng, Jielin Chen, Weimin Zhu, Daping Wang
      Abstract: DNA methylation has emerged as a crucial regulator of chondrocyte dedifferentiation, which severely compromises the outcome of autologous chondrocyte implantation (ACI) treatment for cartilage defects. However, the full-scale DNA methylation profiling in chondrocyte dedifferentiation remains to be determined. Here, we performed a genome-wide DNA methylation profiling of dedifferentiated chondrocytes in monolayer culture and chondrocytes treated with DNA methylation inhibitor 5-azacytidine (5-AzaC). This research revealed that the general methylation level of CpG was increased while the COL-1A1 promoter methylation level was decreased during the chondrocyte dedifferentiation. 5-AzaC could reduce general methylation levels and reverse the chondrocyte dedifferentiation. Surprisingly, the DNA methylation level of COL-1A1 promoter was increased after 5-AzaC treatment. The COL-1A1 expression level was increased while that of SOX-9 was decreased during the chondrocyte dedifferentiation. 5-AzaC treatment up-regulated the SOX-9 expression while down-regulated the COL-1A1 promoter activity and gene expression. Taken together, these results suggested that differential regulation of the DNA methylation level of cartilage-specific genes might contribute to the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.Differential regulation of the DNA methylation level of cartilage-specific genes might contribute the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation.
      PubDate: 2017-01-31T07:47:23.34891-05:0
      DOI: 10.1002/jcp.25486
  • Glioblastoma Multiforme and Adult Neurogenesis in the
           Ventricular-Subventricular Zone: A Review
    • Authors: Claudia Capdevila; Lucía Rodríguez Vázquez, Joaquín Martí
      Abstract: Brain cancers account for
      PubDate: 2017-01-31T07:47:08.85698-05:0
      DOI: 10.1002/jcp.25502
  • Unique Regulatory Mechanisms for the Human Embryonic Stem Cell Cycle
    • Authors: Jennifer J. VanOudenhove; Rodrigo A. Grandy, Prachi N. Ghule, Jane B. Lian, Janet L. Stein, Sayyed K. Zaidi, Gary S. Stein
      Abstract: The cell cycle in pluripotent human embryonic stem cells is governed by unique mechanisms that support unrestricted proliferation and competency for endodermal, mesodermal, and ectodermal differentiation. The abbreviated G1 period with retention of uncompromised fidelity for genetic and epigenetic mechanisms operative in control of proliferation support competency for expansion of the pluripotent cell population that is fundamental for initial stages of development. Regulatory events during the G1 period of the pluripotent cell cycle are decisive for the transition from pluripotency to lineage commitment. Recent findings indicate that a G2 cell cycle pause is present in both endodermal and mesodermal lineage cells, and is obligatory for differentiation to endoderm. J. Cell. Physiol. 9999: 1–4, 2016. © 2016 Wiley Periodicals, Inc.The abbreviated G1 period with retention of uncompromised fidelity for genetic and epigenetic mechanisms operative in control of proliferation support competency for expansion of the pluripotent cell population that is fundamental for initial stages of development. Regulatory events during the G1 period of the pluripotent cell cycle are decisive for the transition from pluripotency to lineage commitment. Recent findings indicate that a G2 cell cycle pause is present in both endodermal and mesodermal lineage cells, and is obligatory for differentiation to endoderm.
      PubDate: 2017-01-31T07:36:36.861307-05:
      DOI: 10.1002/jcp.25567
  • Cytoskeleton Aberrations in Alkaptonuric Chondrocytes
    • Authors: Michela Geminiani; Silvia Gambassi, Lia Millucci, Pietro Lupetti, Giulia Collodel, Lucia Mazzi, Bruno Frediani, Daniela Braconi, Barbara Marzocchi, Marcella Laschi, Giulia Bernardini, Annalisa Santucci
      Abstract: Alkaptonuria (AKU) is an ultra-rare autosomal genetic disorder caused by a defect in the activity of the enzyme homogentisate 1,2-dioxygenase (HGD) that leads to the accumulation of homogentisic acid (HGA) and its oxidized product, benzoquinone acetic acid (BQA), in the connective tissues causing a pigmentation called “ochronosis.” The consequent progressive formation of ochronotic aggregates generate a severe condition of oxidative stress and inflammation in all the affected areas. Experimental evidences have also proved the presence of serum amyloid A (SAA) in several AKU tissues and it allowed classifying AKU as a secondary amyloidosis. Although AKU is a multisystemic disease, the most affected system is the osteoarticular one and articular cartilage is the most damaged tissue. In this work, we have analyzed for the first time the cytoskeleton of AKU chondrocytes by means of immunofluorescence staining. We have shown the presence of SAA within AKU chondrocytes and finally we have demonstrated the co-localization of SAA with three cytoskeletal proteins: actin, vimentin, and β-tubulin. Furthermore, in order to observe the ultrastructural features of AKU chondrocytes we have performed TEM analysis, focusing on the Golgi apparatus structure and, to demonstrate that pigmented areas in AKU cartilage are correspondent to areas of oxidation, 4-HNE presence has been evaluated by means of immunofluorescence. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.The cytoskeleton of alkaptonuric chondrocytes is severely alterated. Serum amyloid A is demonstrated to be present within alkaptonuric chondrocytes and to co-localize with the three main cytoskeletal proteins: actin, vimentin, and β-tubulin. These findings lead to hypothesize a role of SAA in cytoskeleton alterations of alkaptonuric chondrocytes.
      PubDate: 2017-01-31T07:29:41.564847-05:
      DOI: 10.1002/jcp.25500
  • Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine
           Signaling Pathway
    • Authors: Maria M. Barreca; Walter Spinello, Vincenzo Cavalieri, Giuseppina Turturici, Gabriella Sconzo, Punit Kaur, Rosaria Tinnirello, Alexzander A. A. Asea, Fabiana Geraci
      Abstract: Mouse mesoangioblasts are vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll-like receptor 4 (TLR4) and CD91 to stimulate migration. We further demonstrate that Hsp70 has a positive role in regulating matrix metalloproteinase 2 (MMP2) and MMP9 expression and that MMP2 has a more pronounced effect on cell migration, as compared to MMP9. In addition, the analysis of the intracellular pathways implicated in Hsp70 regulated signal transduction showed the involvement of both PI3K/AKT and NF-κB. Taken together, our findings present a paradigm shift in our understanding of the molecular mechanisms that regulate mesoangioblast stem cells ability to traverse the extracellular matrix (ECM). J. Cell. Physiol. 9999: 1–17, 2016. © 2016 Wiley Periodicals, Inc.Extracellular Hsp70 released through extracellular vesicles regulates mesoangioblast migration via the activation of NF-κB and PI3K/AKT pathways.
      PubDate: 2017-01-31T07:19:01.293202-05:
      DOI: 10.1002/jcp.25722
  • Long Non-coding RNAsinTriple Negative Breast Cancer
    • Authors: Qiuhong Wang; Sheng Gao, Haibo Li, Mingming Lv, Cheng Lu
      Abstract: Long non-coding RNAs (lncRNAs) are dysregulated in many cancer types, which are believed to play crucial roles in regulating several hallmarks of cancer biology. Triple Negative Breast Cancer (TNBC) is a very aggressive subtype of normal breast cancer, which has features of negativity for ER, PR and HER2. Great efforts have been made to identify an association between lncRNAs expression profiles and TNBC, and to understand the functional role and molecular mechanism on aberrant-expressed lncRNAs. In this review, we summarized the existed knowledge on the systematics, biology, and function of lncRNAs. The advances from the most recent studies of lncRNAs in the predicament of breast cancer, TNBC, are highlighted, especially the functions of specifically selected lncRNAs. We also discussed the potential value of these lncRNAs in TNBC, providing clues for the diagnosis and treatments of TNBC. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-31T00:06:19.751929-05:
      DOI: 10.1002/jcp.25830
  • High Glucose Alters the Secretome of Mechanically Stimulated
           Osteocyte-like Cells Affecting Osteoclast Precursor Recruitment and
    • Authors: Marta Maycas; María Teresa Portolés, María Concepción Matesanz, Irene Buendía, Javier Linares, María José Feito, Daniel Arcos, María Vallet-Regí, Lilian Plotkin, Pedro Esbrit, Arancha R. Gortázar
      Abstract: Diabetes mellitus (DM) induces bone deterioration, while mechanical stimulation promotes osteocyte-driven bone formation. We aimed to evaluate the interaction of acute exposure (24h) to high glucose (HG) with both the pro-survival effect conferred to osteocytic MLO-Y4 cells and osteoblastic MC3T3-E1 cells by mechanical stimulation and the interaction of these cells with osteoclast precursor RAW264.7 cells. We found that 24h of HG (25 mM) pre-exposure prevented both cell survival and ERK and β-catenin nuclear translocation upon mechanical stimulation by fluid flow (FF) (10 min) in both MLO-Y4 and MC3T3-E1 cells. However, migration of RAW 264.7 cells was inhibited by MLO-Y4 cell-conditioned medium (CM), but not by MC3T3-E1 cell-CM, with HG or FF. This inhibitory effect was associated with consistent changes in VEGF, RANTES, MIP-1α, MIP-1β MCP-1 and GM-CSF in MLO-Y4 cell-CM. RAW264.7 proliferation was inhibited by MLO-Y4 CM under static or HG conditions, but itincreased by FF-CM with or without HG. In addition, both FF and HG abrogated the capacity of RAW 264.7 cells to differentiate into osteoclasts, but in a different manner. Thus, HG-CM in static condition allowed formation of osteoclast-like cells, which were unable to resorb hydroxyapatite. In contrast, FF-CM prevented osteoclastogenesis even in HG condition. Moreover, HG did not affect basal RANKL or IL-6 secretion or their inhibition induced by FF in MLO-Y4 cells. In conclusion, this in vitro study demonstrates that HG exerts disparate effects on osteocyte mechanotransduction, and provides a novel mechanism by which DM disturbs skeletal metabolism through altered osteocyte-osteoclast communication. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-31T00:06:13.20643-05:0
      DOI: 10.1002/jcp.25829
  • HDAC1, HDAC4, and HDAC9 Bind to PC3/Tis21/Btg2 and Are Required for Its
           Inhibition of Cell Cycle Progression and Cyclin D1 Expression
    • Authors: Laura Micheli; Giorgio D'Andrea, Luca Leonardi, Felice Tirone
      Abstract: PC3/Tis21 is a transcriptional cofactor that inhibits proliferation in several cell types, including neural progenitors. Here, we report that PC3/Tis21 associates with HDAC1, HDAC4, and HDAC9 in vivo, in fibroblast cells. Furthermore, when HDAC1, HDAC4, or HDAC9 are silenced in fibroblasts or in a line of cerebellar progenitor cells, the ability of PC3/Tis21 to inhibit proliferation is significantly reduced. Overexpression of HDAC1, HDAC4, or HDAC9 in fibroblasts and in cerebellar precursor cells synergizes with PC3/Tis21 in inhibiting the expression of cyclin D1, a cyclin selectively inhibited by PC3/Tis21. Conversely, the depletion of HDAC1 or HDAC4 (but not HDAC9) in fibroblasts and in cerebellar precursor cells significantly impairs the ability of PC3/Tis21 to inhibit cyclin D1 expression. An analysis of HDAC4 deletion mutants shows that both the amino-terminal moiety and the catalytic domain of HDAC4 associate to PC3/Tis21, but neither alone is sufficient to potentiate the inhibition of cyclin D1 by PC3/Tis21. As a whole, our findings indicate that PC3/Tis21 inhibits cell proliferation in a way dependent on the presence of HDACs, in fibroblasts as well as in neural cells. Considering that several reports have demonstrated that HDACs can act as transcriptional corepressors on the cyclin D1 promoter, our data suggest that the association of PC3/Tis21 to HDACs is functional to recruit them to target genes, such as cyclin D1, for repression of their expression. J. Cell. Physiol. 9999: 1–12, 2016. © 2016 Wiley Periodicals, Inc.By silencing HDAC1, HDAC4, and HDAC9 in fibroblast as well as in neural cells, we reveal that HDAC1, HDAC4, and HDAC9 are each required for the negative control of the cell cycle exerted by PC3/Tis21. Such effect is likely dependent on the ability of HDAC1, HDAC4, and HDAC9 to associate with PC3/Tis21, that we demonstrate here. Moreover, we find that HDAC1 and HDAC4—but not HDAC9—are required for the inhibition exerted by PC3/Tis21 of cyclin D1 expression.
      PubDate: 2017-01-27T06:45:37.185904-05:
      DOI: 10.1002/jcp.25467
  • Advanced Glycation End Products (AGE) Potentiates Cell Death in p53
           Negative Cells via Upregulaion of NF-kappa B and Impairment of Autophagy
    • Authors: Neeharika Verma; Sunil K Manna
      Abstract: Accumulation of advanced glycation end products (AGE) in diabetic patients and ageing people due to excess availability of simple 3- or 4-carbon sugars, is well-known. AGE has multiple deleterious effects including age-related disorders, apoptosis, inflammation and obesity. We have found that AGE increases autophagy but the sustained amount of autophagosomes is observed till 3 days without maturation. It is important to understand the underlying mechanism of AGE-mediated signaling responsible for impairment of autophagy and its correlation to the induction of several adverse effects. We have identified cross talk between autophagy and apoptosis upon AGE stimulation, specifically in p53 negative cells. AGE impairs autophagosomes' clearance in p53 negative cells as observed with an autophagosome maturation blocker- bafilomycinA1 treated cells. This autophagy impairment is well supported by upregulation and overexpression of NF-κB in these p53 negative cells. Autophagy impairment acts as a switch to initiate apoptosis via regulation of NF-κB and its dependent genes. Increase in the expression of NF-κB-dependent NEDD4, an E3 ubiquitin ligase, which targets Beclin1 for cleavage is also evident. Beclin1 interacts with Bcl-2, an anti-apoptotic protein thereby engaging it to facilitate apoptosis upon AGE stimulation. For the first time, we are providing data that NF-κB targeted cell signaling is involved in AGE-mediated autophagy impairment in p53 negative/null cells. The p53 acts antagonistically to prevent this impairment. This study will help to control the AGE-mediated detrimental effects associated with ageing and lysosomal storage disorders. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-27T06:41:25.146761-05:
      DOI: 10.1002/jcp.25828
  • Chronic Infusion of SOD1G93A Astrocyte-Secreted Factors Induces Spinal
           Motoneuron Degeneration and Neuromuscular Dysfunction in Healthy Rats
    • Authors: UN Ramírez-Jarquín; F Rojas, B van Zundert, R Tapia
      Abstract: Amyotrophic lateral sclerosis is a fatal neurodegenerative disease and studies in vitro show that motoneuron degeneration is triggered by non-cell-autonomous mechanisms. However, whether soluble toxic factor(s) released by mutant superoxide dismutase 1 (SOD1) expressing astrocytes induces death of motoneurons and leads to motor dysfunction in vivo is not known. To directly test this, healthy adult rats were treated with conditioned media derived from primary mouse astrocytes (ACM) that express human (h) SOD1G93A (ACM-hG93A) via chronic osmotic pump infusion in the lumbar spinal cord. Controls included ACM derived from transgenic mice expressing hSOD1WT (ACM-hWT) or non-transgenic mouse SOD1WT (ACM-WT) astrocytes. Rats chronically infused with ACM-hG93A started to develop motor dysfunction at 8 days, as measured by rotarod performance. Additionally, immunohistochemical analyses at day 16 revealed reactive astrogliosis and significant loss of motoneurons in the ventral horn of the infused region. Controls did not show significant motor behavior alterations or neuronal damage. Thus, we demonstrate that factors released in vitro from astrocytes derived from ALS mice cause spinal motoneuron death and consequent neuromuscular dysfunction in vivo. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-27T06:31:40.942632-05:
      DOI: 10.1002/jcp.25827
  • Distinctive Expression Pattern of Cystathionine-β-Synthase and
           Cystathionine-γ-Lyase Identifies Mesenchymal Stromal Cells Transition to
           Mineralizing Osteoblasts
    • Authors: Laura Gambari; Gina Lisignoli, Elena Gabusi, Cristina Manferdini, Francesca Paolella, Anna Piacentini, Francesco Grassi
      Abstract: Mesenchymal stromal cells (MSCs) are key players in the repair or regeneration of the damaged bone tissue. However, heterogeneity exists between MSCs derived from different donors in their bone formation ability both in vitro and in vivo. The identification of markers defining MSCs with different functional phenotypes is fundamental to maximize their clinical potential. In our previous in vivo study, impaired expression in MSCs of cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE), the two key enzymes in the catabolic pathway of homocysteine, was associated to decreased bone formation and to the onset of osteoporosis in mice. Here, we investigated whether osteogenic differentiation of human MSCs (hMSCs) modulates the expression of CBS and CSE. The expression of CBS and CSE was also assessed during chondrogenesis to confirm the specificity of their expression during osteogenesis. hMSCs displayed a heterogeneous mineralizing capacity between donors (70% of the samples mineralized, while 30% did not mineralize). Inducible expression of CBS and CSE was found to be associated with a mineralizing phenotype in hMSCs. In particular, up-regulation of CSE was restricted to hMSCs undergoing mineralization. During chondrogenesis, CBS was significantly up-regulated while CSE expression was not affected. Ex-vivo findings confirmed that mature h-osteoblasts (hOBs) show consistently higher expression of CBS and CSE than hMSCs.Our data provide the first evidence that the expression of CBS and CSE in hMSCs closely correlates with the transition of hMSCs towards the osteoblastic phenotype and that CSE may constitute a novel marker of osteogenic differentiation. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-25T06:30:35.46048-05:0
      DOI: 10.1002/jcp.25825
  • Nanotubes Impregnated Human Olfactory Bulb Neural Stem Cells Promote
           Neuronal Differentiation in Trimethyltin-induced Neurodegeneration Rat
    • Authors: HE Marei; AA Elnegiry, A Zaghloul, A Althani, N Afifi, A Abd-Elmaksoud, A Farag, S Lashen, S Rezk, Z Shouman, C Cenciarelli, Anwarul Hasan
      Abstract: Neural stem cells (NSCs) are multipotent self-renewing cells that could be used in cellular-based therapy for a wide variety of neurodegenerative diseases including Alzheimer's diseases (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS). Being multipotent in nature, they are practically capable of giving rise to major cell types of the nervous tissue including neurons, astrocytes and oligodendrocytes. This is in marked contrast to neural progenitor cells which are committed to a specific lineage fate. In previous studies, we have demonstrated the ability of NSCs isolated from human olfactory bulb (OB) to survive, proliferate, differentiate, and restore cognitive and motor deficits associated with AD, and PD rat models, respectively. The use of carbon nanotubes (CNTs) to enhance the survivability and differentiation potential of NSCs following their in vivo engraftment have been recently suggested. Here, in order to assess the ability of CNTs to enhance the therapeutic potential of human OBNSCs for restoring cognitive deficits and neurodegenerative lesions, we co-engrafted CNTs and human OBNSCs in TMT-neurodegeneration rat model. The present study revealed that engrafted human OBNSCS-CNTs restored cognitive deficits, and neurodegenerative changes associated with TMT-induced rat neurodegeneration model. Moreover, the CNTs seemed to provide a support for engrafted OBNSCs, with increasing their tendency to differentiate into neurons rather than into glia cells. The present study indicate the marked ability of CNTs to enhance the therapeutic potential of human OBNSCs which qualify this novel therapeutic paradigm as a promising candidate for cell-based therapy of different neurodegenerative diseases. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-25T06:30:29.550501-05:
      DOI: 10.1002/jcp.25826
  • Membrane Ion Channels and Receptors in Animal Lifespan Modulation
    • Authors: Yi Sheng; Lanlan Tang, Lijun Kang, Rui Xiao
      Abstract: Acting in the interfaces between environment and membrane compartments, membrane ion channels and receptors transduce various physical and chemical cues into downstream signaling events. Not surprisingly, these membrane proteins play essential roles in a wide range of cellular processes such as sensory perception, synaptic transmission, cellular growth and development, fate determination, and apoptosis. However, except insulin and insulin-like growth factor receptors, the functions of membrane receptors in animal lifespan modulation have not been well appreciated. On the other hand, although ion channels are popular therapeutic targets for many age-related diseases, their potential roles in aging itself are largely neglected. In this review, we will discuss our current understanding of the conserved functions and mechanisms of membrane ion channels and receptors in the modulation of lifespan across multiple species including C. elegans, Drosophila, mouse, and human. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-25T06:05:42.831459-05:
      DOI: 10.1002/jcp.25824
  • Cytokine Networks and Their Association with Helicobacter pylori Infection
           in Gastric Carcinoma
    • Authors: Vahid Bagheri; Bahram Memar, Amir Abbas Momtazi, Amirhossein Sahebkar, Mohammad Reza Abbaszadegan, Mehran Gholamin
      Abstract: Cytokine networks as dynamic networks are pivotal aspects of tumor immunology, especially in gastric cancer (GC), in which infection, inflammation, and antitumor immunity are key elements of disease progression. In this review, we describe functional roles of well-known GC-modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)-induced inflammation in GC and discuss how HP-induced chronic inflammation can lead to the induction of stem cell hyperplasia, morphological changes in gastric mucosa and GC development. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-25T06:05:41.5194-05:00
      DOI: 10.1002/jcp.25822
  • Molecular, Biophysical, and Pharmacological Properties of
           Calcium-Activated Chloride Channels
    • Authors: Mohammad Amin Kamaleddin
      Abstract: Calcium-activated chloride channels (CaCCs) are a family of anionic transmembrane ion channels. They are mainly responsible for the movement of Cl− and other anions across the biological membranes and are widely expressed in different tissues. Due to the fact that Cl− flow into or out of the cell plays a crucial role in hyperpolarizing or depolarizing the cells, respectively, the impact of intracellular Ca2+ concentration on these channels is attracting a lot of attentions. After summarizing the molecular, biophysical, and pharmacological properties of CaCCs, the role of CaCCs in normal cellular functions will be discussed, and I will emphasize how dysregulation of CaCCs in pathological conditions can account for different diseases. A better understanding of CaCCs and a pivotal regulatory role of Ca2+ can shed more light on the therapeutic strategies for different neurological disorders that arise from chloride dysregulation, such as asthma, cystic fibrosis, and neuropathic pain. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-25T06:05:24.578298-05:
      DOI: 10.1002/jcp.25823
  • Understanding the Role of Structural Integrity and Differential Expression
           of Integrin Profiling to Identify Potential Therapeutic Targets in Breast
    • Authors: Vishal Das; Gazal Kalyan, Saugata Hazra, Mintu Pal
      Abstract: Breast cancer is found to be the most prevalent neoplasm in women worldwide. Despite the function of physically tethering cells to the matrix, transmembrane protein integrins are crucially involved in diverse cellular functions such as cell differentiation, proliferation, invasion, migration and metastasis. Dysregulation of integrins and their interactions with the cells and their microenvironment can trigger several signalling cues that determine the cell fate decision. In this review we spotlight all pre-existing integrin molecules, their structure, molecular interactions motifs, and function through several cross talks with kinase receptors. We also discuss the role of these integrins as potential prognostic and therapeutic targets and also in the regulation of breast cancer cells differentiation. Understanding of integrin structure and their motifs for ligand interactions in this context will enable the development of new therapeutic approaches to sensitize the tumours and their microenvironment to conventional therapy and overall suppress their metastatic phenotype. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-24T22:50:54.56334-05:0
      DOI: 10.1002/jcp.25821
  • A Systems Approach to Physiologic Evolution: From Micelles to
    • Authors: John S. Torday; William B. Miller
      Abstract: A systems approach to evolutionary biology offers the promise of an improved understanding of the fundamental principles of life through the effective integration of many biologic disciplines. It is presented that any critical integrative approach to evolutionary development involves a paradigmatic shift in perspective, more than just the engagement of a large number of disciplines. Critical to this differing viewpoint is the recognition that all biological processes originate from the unicellular state and remain permanently anchored to that phase throughout evolutionary development despite their macroscopic appearances. Multicellular eukaryotic development can therefore be viewed as a series of connected responses to epiphenomena that proceeds from that base in continuous iterative maintenance of collective cellular homeostatic equipoise juxtaposed against an ever-changing and challenging environment. By following this trajectory of multicellular eukaryotic evolution from within unicellular First Principles of Physiology forward, the mechanistic nature of complex physiology can be identified through a step-wise analysis of a continuous arc of vertebrate evolution based upon serial exaptations. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-23T07:00:51.260652-05:
      DOI: 10.1002/jcp.25820
  • Interplay between the miRNome and the Epigenetic Machinery: Implications
           in Health and Disease
    • Authors: Shagun Poddar; Devesh Kesharwani, Malabika Datta
      Abstract: Epigenetics refers to functionally relevant genomic changes that do not involve changes in the basic nucleotide sequence. Majorly, these are of two types- DNA methylation and histone modifications. Small RNA molecules called miRNAs are often thought to mediate post-transcriptional epigenetic changes by mRNA degradation or translational attenuation. While DNA methylation and histone modifications have their own independent effects on various cellular events, several reports are suggestive of an obvious interplay between these phenomena and the miRNA regulatory program within the cell. Several miRNAs like miR-375, members of miR-29 family, miR-34, miR-200 and others are regulated by DNA methylation and histone modifications in various types of cancers and metabolic diseases. On the other hand, miRNAs like miR-449a, miR-148, miR-101, miR-214 and miR-128 target the members of epigenetic machinery and their dysregulation leads to diverse cellular aberrations. In spite of being independent cellular events, emergence of reports that suggest a connection between DNA methylation, histone modification and miRNA function in several diseases indicate that this connecting axis offers a valuable target with great therapeutic potential that might be exploited for disease management. We review the current status of crosstalk between the major epigenetic modifications and the miRNA machinery and discuss this in the context of health and disease. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-23T06:55:34.66382-05:0
      DOI: 10.1002/jcp.25819
  • Intracellular Transcytosis of Albumin in Glomerular Endothelial Cells
           After Endocytosis Through Caveolae
    • Authors: Takahito Moriyama; Kayo Sasaki, Kazunori Karasawa, Keiko Uchida, Kosaku Nitta
      Abstract: Background/AimsWe previously described albumin endocytosis through caveolae in human renal glomerular endothelial cells (HRGECs). This suggested a new albumin transcytosis pathway, in addition to the fenestral pathway. As a next step, we investigated albumin transcytosis in HRGECs after caveolar endocytosis.Methods and ResultsHRGECs were incubated with Alexa Fluor 488-labeled bovine serum albumin from 0 to 360 minutes. Next, markers for endosomes, endoplasmic reticulum (ER), golgi apparatus (GA), lysosomes and proteasomes and Fc receptors, microtubules and actin were monitored by immunofluorescence. Labeled albumin co-localization with endosomes was gradually and significantly increased and it was significantly higher than with the other markers at any timepoint. Albumin, placed on inside of the Transwell membrane, diffused through HRGEC monolayers during a 360-min incubation period. This transportation of albumin through HRGECs was inhibited by methyl beta cyclodextrin (MBCD), a caveolae disrupting agent. MBCD also decreased albuminuria, causing decreased caveolin-1 expression on glomerular capillaries, in puromycin aminonucleoside induced nephrotic mice.ConclusionAlbumin transcytosis depends on early endosomes, but not on other organelles, Fc receptors or cytoskeletal components. Caveolae disruption prevented albumin transportation through HRGECs and decreased albuminuria in nephrotic mice. This newly described caveolae-dependent albumin pathway through glomerular endothelial cells is a potential pathogenetic mechanism for albuminuria, independent of the fenestrae. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-23T06:55:30.142115-05:
      DOI: 10.1002/jcp.25817
  • SUMO-modified Insulin-Like Growth Factor 1 Receptor (IGF-1R) Increases
           Cell Cycle Progression and Cell Proliferation
    • Authors: Yingbo Lin; Hongyu Liu, Ahmed Waraky, Felix Haglund, Prasoon Agarwal, Helena Jernberg-Wiklund, Dudi Warsito, Olle Larsson
      Abstract: Increasing number of studies have shown nuclear localization of the insulin-like growth factor 1 receptor (nIGF-1R) in tumor cells and its links to adverse clinical outcome in various cancers. Any obvious cell physiological roles of nIGF-1R have, however, still not been disclosed. Previously, we reported that IGF-1R translocates to cell nucleus and modulates gene expression by binding to enhancers, provided that the receptor is SUMOylated. In this study, we constructed stable transfectants of wild type IGF1R (WT) and triple-SUMO-site-mutated IGF1R (TSM) using igf1r knockout mouse fibroblasts (R-). Cell clones (R-WT and R-TSM) expressing equal amounts of IGF-1R were selected for experiments. Phosphorylation of IGF-1R, Akt and Erk upon IGF-1 stimulation was equal in R-WT and R-TSM. WT was confirmed to enter nuclei. TSM did also undergo nuclear translocation, although to a lesser extent. This may be explained by that TSM heterodimerizes with insulin receptor, which is known to translocate to cell nuclei. R-WT proliferated substantially faster than R-TSM, which did not differ significantly from the empty vector control. Upon IGF-1 stimulation G1-S-phase progression of R-WT increased from 12 to 38%, compared to 13 to 20% of R-TSM. The G1-S progression of R-WT correlated with increased expression of cyclin D1, A and CDK2, as well as downregulation of p27. This suggests that SUMO-IGF-1R affects upstream mechanisms that control and coordinate expression of cell cycle regulators. Further studies to identify such SUMO-IGF-1R dependent mechanisms seem important. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-23T06:55:27.894536-05:
      DOI: 10.1002/jcp.25818
  • Mesenchymal Stem Cells Improves Survival in LPS-induced Acute Lung Injury
           Acting through Inhibition of NETs Formation
    • Authors: Leonardo Pedrazza; Aline Andrea Cunha, Carolina Luft, Nailê Karine Nunes, Felipe Schimitz, Rodrigo Benedetti Gassen, Ricardo Vaz Breda, Marcio Vinícius Fagundes Donadio, Angela Terezinha de Souza Wyse, Paulo Marcio Condessa Pitrez, Jose Luis Rosa, Jarbas Rodrigues de Oliveira
      Abstract: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are syndromes of acute hypoxemic respiratory failure resulting from a variety of direct and indirect injuries to the gas exchange parenchyma of the lungs. During the ALI, we have an increase release of proinflammatory cytokines and high reactive oxigen species (ROS) formation. These factors are responsible for the release and activation of neutrophil-derived proteases and the formation of neutrophil extracellular traps (NETs). The excessive increase in the release of NETs cause damage to lung tissue. Recent studies have studies involving the administration of mesenchymal stem cells (MSCs) for the treatment of experimental ALI has shown promising results. In this way, the objective of our study is evaluate the ability of MSCs, in a lipopolysaccharide (LPS)-induced ALI model, to reduce inflammation, oxidative damage and consequently decrease the release of NETs. Mice were submitted lung injury induced by intratracheal instillation of LPS and subsequently treated or not with MSCs. Treatment with MSCs was able to modulate pulmonary inflammation, decrease oxidative damage and reduce the release of NETs. These benefits from treatment are evident when we observe a significant increase in the survival curve in the treated animals. Our results demonstrate that MSCs treatment is effective for the treatment of ALI. For the first time it is described that MSCs can reduce the formation of NETs and an experimental model of ALI. This finding is directly related to these cells modulate the inflammatory response and oxidative damage in the course of the pathology. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-23T06:55:24.112655-05:
      DOI: 10.1002/jcp.25816
  • Lipid Profiling of Parkin-Mutant Human Skin Fibroblasts
    • Authors: Simona Lobasso; Paola Tanzarella, Daniele Vergara, Michele Maffia, Tiziana Cocco, Angela Corcelli
      Abstract: Parkin mutations are a major cause of early-onset Parkinson's disease (PD). The impairment of protein quality control system together with defects in mitochondria and autophagy process are consequences of the lack of parkin, which leads to neurodegeneration. Little is known about the role of lipids in these alterations of cell functions.In the present study parkin-mutant human skin primary fibroblasts have been considered as cellular model of PD to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid composition were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) and thin-layer chromatography (TLC). In parallel, we have performed direct MALDI-TOF/MS lipid analyses of intact fibroblasts by skipping lipid extraction steps.Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin-mutant fibroblasts. The detected higher level of gangliosides, phosphatidylinositol and phosphatidylserine could be linked to dysfunction of autophagy and mitochondrial turnover; in addition, the lysophosphatidylcholine increase could represent the marker of neuroinflammatory state, a well-known component of Parkinson's disease. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-21T06:20:30.606152-05:
      DOI: 10.1002/jcp.25815
  • VSL#3 Probiotic Differently Influence IEC-6 Intestinal Epithelial Cell
           Status and Function
    • Authors: Benedetta Cinque; Cristina La Torre, Francesca Lombardi, Paola Palumbo, Zoran Evtoski, Silvano Jr Santini, Stefano Falone, Annamaria Cimini, Fernanda Amicarelli, Maria Grazia Cifone
      Abstract: The data here reported introduce the wound-healing assay as a tool for testing probiotics aimed at protecting gastrointestinal mucosal surfaces and to verify the consistency of their manufacturing. At the scope, we compared the in vitro effects of two multi-strain high concentration formulations both commercialized under the same brand VSL#3 but sourced from different production sites (USA and Italy) on a non-transformed small-intestinal epithelial cell line, IEC-6. The effects on cellular morphology, viability, migration, and H2O2-induced damage, were assessed before and after the treatment with both VSL#3 formulations. While the USA-sourced product (“USA-made”) VSL#3 did not affect monolayer morphology and cellular density, the addition of bacteria from the Italy-derived product (“Italy-made”) VSL#3 caused clear morphological cell damage and strongly reduced cellularity. The treatment with “USA-made” lysate led to a higher rate of wounded monolayer healing, while the addition of “Italy-made” bacterial lysate did not influence the closure rate as compared to untreated cells. While lysates from “USA-made” VSL#3 clearly enhanced the formation of elongated and aligned stress fibers, “Italy-made” lysates had not similar effect. “USA-made” lysate was able to cause a total inhibition of H2O2-induced cytotoxic effect whereas “Italy-made” VSL#3 lysate was unable to protect IEC-6 cells from H2O2-induced damage. ROS generation was also differently influenced, thus supporting the hypotesis of a protective action of “USA-made” VSL#3 lysates, as well as the idea that “Italy-made” formulation was unable to prevent significantly the H2O2-induced oxidative stress. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-21T06:20:29.468969-05:
      DOI: 10.1002/jcp.25814
  • The Prognostic and Therapeutic Application of microRNAs in Breast Cancer:
           Tissue and Circulating microRNAs
    • Authors: Afsane Bahrami; Seyed Amir Aledavoud, Kazem Anvari, Seyed Mahdi Hassanian, Mina Maftouh, Ali Yaghobzade, Omid Salarzaee, Soodabeh ShahidSales, Amir Avan
      Abstract: Breast cancer is the second most common malignancy diagnosed in women, supporting the need for identification of novel prognostic and diagnostic biomarkers. Recently, microRNAs have emerged as molecular regulators that can have key roles in pathogenesis and progression of different malignancies, including breast cancer. Micro-RNAs can be circulated in body fluid, suggesting their values as non-invasive marker. There is growing body of evidence showing the aberrant activation of some known circulating miRNAs, e.g. let-151a, miR-21, miR-155, miR-,145 miR-18a, miR-16 as well as tissue specific-miRNAs, e.g. miR-182, miR-145, miR-21, miR-155/154, miR-203, miR-213, miR-7 in patients affected by breast cancer. In addition, there is growing body of evidences on the value of miRNAs to be associated with drug-resistance, suggesting their values as a potential approach to overcome chemo-resistance. Attuned with these facts, this review highlights recent preclinical and clinical investigation performed on tissue-specific miRNAs and circulating as novel promising biomarkers for detection of patients at early stages, prediction of prognosis, and monitoring of the patients in response to therapy. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-21T06:20:23.684844-05:
      DOI: 10.1002/jcp.25813
  • Serotonin Transporter Protects the Placental Cells Against Apoptosis in
           Caspase 3-Independent Pathway
    • Authors: Coedy Hadden; Tariq Fahmi, Anthonya Cooper, Alena V Savenka, Vladimir V. Lupashin, Drucilla J. Roberts, Luc Maroteaux, Sylvie Hauguel-de Mouzon, Fusun Kilic
      Abstract: Serotonin (5-HT) and its specific transporter, SERT play important roles in pregnancy. Using placentas dissected from 18d gestational SERT-knock out (KO), peripheral 5-HT (TPH1)-KO and wild-type (WT) mice, we explored the role of 5-HT and SERT in placental functions in detail. An abnormal thick band of fibrosis and necrosis under the giant cell layer in SERT-KO placentas appeared only moderately in TPH1-KO and minimally present in WT placentas. The majority of the changes were located at the junctional zone of the placentas in SERT. The etiology of these findings was tested with TUNEL assays. The placentas from SERT-KO and TPH1-KO showed 49- and 8-fold increase in TUNEL-positive cells without a concurrent change in the DNA repair or cell proliferation compared to WT placentas. While the proliferation rate in the embryos of TPH1-KO mice was 16-fold lower than the rate in gestational age matched embryos of WT or SERT-KO mice. These findings highlight an important role of continuous 5-HT signaling on trophoblast cell viability. SERT may contribute to protecting trophoblast cells against cell death via terminating the 5-HT signaling which changes cell death ratio in trophoblast as well as proliferation rate in embryos. However, the cell death in SERT-KO placentas is in caspase 3-independent pathway. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-21T06:10:53.610234-05:
      DOI: 10.1002/jcp.25812
  • LSP1, a Responsive Protein from Meyerozyma guilliermondii, Elicits Defence
           Response and Improves Glycyrrhizic Acid Biosynthesis in Glycyrrhiza
           uralensis Fisch Adventitious Roots
    • Authors: Juan Wang; Jianli Li, Jing Li, Jinxin Li, Shujie Liu, Wenyuan Gao
      Abstract: This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the Meyerozyma guilliermondii since the excellent induction. The contents of total flavonoids (3.46 mg · g−1), glycyrrhizic acid (0.41 mg · g−1), glycyrrhetinic acid (0.41 mg · g−1) and polysaccharide (94.49 mg · g−1) in adventitious root peaked at LSP1 group, which were 1.6, 3.4, 2.4, 2.0-fold that of control, respectively. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen-activated protein kinases and extremely up-regulated the expression of defense-related genes and functional genes involved in glycyrrhizic acid biosynthesis. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-20T00:30:26.267671-05:
      DOI: 10.1002/jcp.25811
  • Mechanisms Involved in Enhancement of Matrix Metalloproteinase-9
           Expression in Macrophages by Interleukin-33
    • Authors: Wataru Ariyoshi; Toshinori Okinaga, Wichida Chaweewannakorn, Sumio Akifusa, Tatsuji Nisihara
      Abstract: Endothelial transmigration of macrophages is accomplished by matrix metalloproteinase (MMP)-induced degradation of the basement membrane and extracellular matrix components. Macrophages up-regulate MMP-9 expression and secretion upon immunological challenges and require its activity for migration during inflammatory responses. Interleukin (IL)-33 is a recently discovered pro-inflammatory cytokine that belongs to the IL-1 family. The aim of this study was to elucidate the mechanisms underlying IL-33-induced MMP-9 expression in the mouse monocyte/macrophage line RAW264.7. IL-33 increased MMP-9 mRNA and protein expression in RAW264.7 cells. Blockage of IL-33-IL-33 receptor (ST2L) binding suppressed IL-33-mediated induction of MMP-9. IL-33 induced phosphorylation and nuclear translocation of extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-kappa B (NF-κB). Chromatin immunoprecipitation indicated that IL-33 increased c-fos recruitment to the MMP-9 promoter. Reporter assay findings also revealed that IL-33 stimulated the transcriptional activity of activator protein 1 (AP-1). Pre-treatment of the cells with a specific inhibitor of ERK1/2 and NF-κB attenuated the IL-33-induced activation of AP-1 subunits, transcriptional activity of AP-1, and expression of MMP-9. We also demonstrated that ERK-dependent activation of cAMP response element binding protein (CREB) is a key step for AP-1 activation by IL-33. These results indicate an essential role of ERK/CREB and NF-κB cascades in the induction of MMP-9 in monocytes/macrophages through AP-1 activation. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-20T00:26:06.461517-05:
      DOI: 10.1002/jcp.25809
  • STIM-1 and ORAI-1 Channel Mediate Angiotensin-II-Induced Expression of
           Egr-1 in Vascular Smooth Muscle Cells
    • Authors: Estelle R. Simo-Cheyou; Ju Jing Tan, Ryszard Grygorczyk, Ashok K. Srivastava
      Abstract: An upregulation of Egr-1 expression has been reported in models of atherosclerosis and intimal hyperplasia and, various vasoactive peptides and growth promoting stimuli have been shown to induce the expression of Egr-1 in vascular smooth muscle cells (VSMC). Angiotensin-II (Ang-II) is a key vasoactive peptide that has been implicated in the pathogenesis of vascular diseases. Ang-II elevates intracellular Ca2+ through activation of the store-operated calcium entry (SOCE) involving an inositol-3-phosphate receptor (IP3R)-coupled depletion of endoplasmic reticular Ca2+ and a subsequent activation of the stromal interaction molecule 1 (STIM-1) /Orai-1 complex. However, the involvement of IP3R/STIM-1/Orai-1-Ca2+-dependent signaling in Egr-1 expression in VSMC remains unexplored. Therefore, in the present studies, we have examined the role of Ca2+ signaling in Ang-II-induced Egr-1 expression in VSMC and investigated the contribution of STIM-1 or Orai-1 in mediating this response. 2-aminoethoxydiphenyl borate (2-APB), a dual non-competitive antagonist of IP3R and inhibitor of SOCE, decreased Ang-II-induced Ca2+ release and attenuated Ang-II-induced enhanced expression of Egr-1 protein and mRNA levels. Egr-1 upregulation was also suppressed following blockade of calmodulin and CaMKII. Furthermore, RNA interference-mediated depletion of STIM-1 or Orai-1 attenuated Ang-II-induced Egr-1 expression as well as Ang-II-induced phosphorylation of ERK1/2 and CREB. In addition, siRNA-induced silencing of CREB resulted in a reduction in the expression of Egr-1 stimulated by Ang-II. In summary, our data demonstrate that Ang-II-induced Egr-1 expression is mediated by STIM-1/Orai-1/Ca2+-dependent signaling pathways in A-10 VSMC. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-20T00:25:51.268573-05:
      DOI: 10.1002/jcp.25810
  • Cover Image, Volume 232, Number 5, May 2017
    • Authors: Toshiaki Tanaka; Kaoru Goto, Mitsuyoshi Iino
      Abstract: Cover: The cover image, by Toshiaki Tanaka, Kaoru Goto, and Mitsuyoshi Iino, is based on the Review Article Diverse Functions and Signal Transduction of the Exocyst Complex in Tumor Cells,
      DOI 10.1002/jcp.25619
      PubDate: 2017-01-19T10:01:01.264005-05:
  • M2-like Macrophages Induce Colon Cancer Cell Invasion via Matrix
    • Authors: Katyayni Vinnakota; Yuan Zhang, Benson Chellakkan Selvanesan, Geriolda Topi, Tavga Salim, Janna Sand-Dejmek, Gunilla Jönsson, Anita Sjölander
      Abstract: The inflammatory milieu plays an important role in colon cancer development and progression. Previously, we have shown that tumor-associated macrophages (TAMs), an important component of the tumor microenvironment, are enriched in tumors compared with normal tissue and confer a poorer prognosis. In the present study, we found that matrix metallopeptidase-9 (MMP-9), which degrades extracellular matrix proteins, was increased in biopsies from colon cancer patients and in mouse xenografts with SW480 cell-derived tumors. SW480 colon cancer cells exposed to M2-like macrophage conditioned medium (M2-medium) exhibited increased MMP-9 mRNA, protein expression and gelatinase activity. A similar effect was obtained by the addition of tumor necrosis factor-α (TNFα) and leukotriene D4 (LTD4). MMP-9 expression and activity were reduced by a TNFα blocking antibody adalimumab and a cysteinyl leukotriene receptor 1 (CysLTR1, the receptor for LTD4) antagonist montelukast. M2-medium also induced changes in the epithelial–mesenchymal transition (EMT) markers E-cadherin, β-catenin, vimentin, and snail in SW480 cells. We also found that both M2-medium and TNFα and LTD4 induced stabilization/nuclear translocation of β-catenin. Furthermore, we also observed an elongated phenotype that may indicate increased invasiveness, as confirmed in a collagen I invasion assay. M2-medium increased the invasive ability, and a similar effect was also obtained by the addition of TNFα and LTD4. The specific MMP inhibitor I or adalimumab and montelukast reduced the number of invasive cells. In conclusion, our findings show that M2-medium enriched in TNFα and LTD4 promote colon cancer cell invasion via MMP-9 expression and activation and the induction of EMT. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-18T06:50:58.52808-05:0
      DOI: 10.1002/jcp.25808
  • Subclinical Alteration of the Cervical-Vaginal Microbiome in Women with
           Idiopathic Infertility
    • Authors: Giuseppina Campisciano; Fiorella Florian, Angela D'Eustacchio, David Stanković, Giuseppe Ricci, Manola Comar
      Abstract: Biomarkers have a wide application in research and clinic, they help to choose the correct treatment for diseases. Recent studies, addressing the vaginal microbiome using next generation sequencing (NGS), reported the involvement of bacterial species in infertility. We compared the vaginal microbiome of idiopathic infertility diagnosed women with that of healthy, non-idiopathic infertile and bacterial vaginosis affected women, to identify bacterial species suitable as biomarkers. Information on microorganisms was obtained from the V3-16S rDNA sequencing of cervical-vaginal fluids of 96 women using the Ion Torrent platform. Data were processed with QIIME and classified against the Vaginal 16S rDNA Reference Database. The analysis revealed a significant beta-diversity variation (p 
      PubDate: 2017-01-18T06:50:47.015254-05:
      DOI: 10.1002/jcp.25806
  • Impact of Lysosomal Storage Disorders on Biology of Mesenchymal Stem
           Cells: Evidences from In Vitro Silencing of Glucocerebrosidase (GBA) and
           Alpha-Galactosidase A (GLA) Enzymes
    • Authors: T Squillaro; I Antonucci, N Alessio, A Esposito, M Cipollaro, MAB Melone, G Peluso, L Stuppia, U Galderisi
      Abstract: Lysosomal storage disorders (LDS) comprise a group of rare multisystemic diseases resulting from inherited gene mutations that impair lysosomal homeostasis. The most common LSDs, Gaucher disease (GD) and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha-galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected. Among stem cells, mesenchymal stem cells (MSCs) are a commonly investigated population given their role in hematopoiesis and the homeostatic maintenance of many organs and tissues. Since the impairment of MSC functions could pose profound consequences on body physiology, we evaluated whether GBA and GLA silencing could affect the biology of MSCs isolated from bone marrow and amniotic fluid. Those cell populations were chosen given the former's key role in organ physiology and the latter's intriguing potential as an alternative stem cell model for human genetic disease. Our results revealed that GBA and GLA deficiencies prompted cell cycle arrest along with the impairment of autophagic flux and an increase of apoptotic and senescence cell percentages. Moreover, an increase in ataxia–telangiectasia-mutated staining 1 h after oxidative stress induction and a return to basal level at 48 h, along with persistent gamma-H2AX staining, indicated that MSCs properly activated DNA repair signaling, though some damages remained unrepaired. Our data therefore suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-18T06:50:36.272131-05:
      DOI: 10.1002/jcp.25807
  • THE Role of Various Peroxisome Proliferator-Activated Receptors and Their
           Ligands in Clinical Practice
    • Authors: Giuseppe Derosa; Amirhossein Sahebkar, Pamela Maffioli
      Abstract: Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors involved in several physiological processes including modulation of cellular differentiation, development, metabolism of carbohydrates, lipids, proteins, and tumorigenesis.The aim of this review is to examine how different PPAR ligands act, and discuss their use in clinical practice.PPAR ligands have a lot of effects and applications in clinical practice. Some PPAR ligands such as fibrates (PPAR-α ligands) are currently used for the treatment of dyslipidemia, while pioglitazone and rosiglitazone (PPAR-γ ligands) are anti-diabetic and insulin-sensitizing agents. Regarding new generation drugs, acting on both α/γ, β /δ, or α/δ receptors simultaneously, preliminary data on PPAR-α/γ dual agonists revealed a positive effect on lipid profile, blood pressure, atherosclerosis, inflammation and anti-coagulant effects, while the overexpression of PPAR-β /δ seems to prevent obesity and to decrease lipid storage in cardiac cells. Finally, PPAR-α/δ dual agonist induces resolution of nonalcoholic steatohepatitis without fibrosis worsening. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-18T06:45:25.380577-05:
      DOI: 10.1002/jcp.25804
  • Inhibitory Effect of Blue Light Emitting Diode on Migration and Invasion
           of Cancer Cells
    • Authors: Phil-Sun Oh; Hyun-Soo Kim, Eun-Mi Kim, Hyosook Hwang, Hyang Hwa Ryu, SeokTae Lim, Myung-Hee Sohn, Hwan-Jeong Jeong
      Abstract: The aim of this study was to determine the effects and molecular mechanism of blue light emitting diode (LED) in tumor cells. A migration and invasion assay for the metastatic behavior of mouse colon cancer CT-26 and human fibrosarcoma HT-1080 cells was performed. Cancer cell migration-related proteins were identified by obtaining a 2-dimensional gel electrophoresis (2-DE) in total cellular protein profile of blue LED-irradiated cancer cells, followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis of proteins. Protein levels were examined by immunoblotting. Irradiation with blue LED inhibited CT-26 and HT-1080 cell migration and invasion. The anti-metastatic effects of blue LED irradiation were associated with inhibition of matrix metalloproteinase (MMP)-2 and MMP-9 expression. P38 MAPK phosphorylation was increased in blue LED-irradiated CT-26 and HT-1080 cells, but was inhibited after pretreatment with SB203580, a specific inhibitor of p38 MAPK. Inhibition of p38 MAPK phosphorylation by SB203580 treatment increased number of migratory cancer cells in CT-26 and HT-1080 cells, indicating that blue LED irradiation inhibited cancer cell migration via phosphorylation of p38 MAPK. Additionally blue LED irradiation of mice injected with CT-26 cells expressing luciferase decreased early stage lung metastasis compared to untreated control mice. These results indicate that blue LED irradiation inhibits cancer cell migration and invasion in vitro and in vivo. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-18T06:45:22.858289-05:
      DOI: 10.1002/jcp.25805
  • MicroRNAs: Potential Candidates for Diagnosis and Treatment of Colorectal
    • Authors: Abdullah Moridikia; Hamed Mirzaei, Amirhossein Sahebkar, Jafar Salimian
      Abstract: Colorectal cancer (CRC) is known as the third common cancer worldwide and an important public health problem in different populations. Several genetics and environmental risk factors are involved in the development and progression of CRC including chromosomal abnormalities, epigenetic alterations, and unhealthy lifestyle. Identification of risk factors and biomarkers could lead to a better understanding of molecular pathways involved in CRC pathogenesis. MicroRNAs (miRNAs) are important regulatory molecules which could affect a variety of cellular and molecular targets in CRC. A large number of studies have indicatedderegulations of some known tissue-specific miRNAs, e.g. miR-21, miR-9, miR-155, miR-17, miR-19, let-7 and miR-24 as well as circulating miRNAs, e.g. miR-181b, miR-21, miR-183, let-7g, miR-17 and miR-126, in patients with CRC. In the current review, we focus on the findings of preclinical and clinical studies performed on tissue-specific and circulating miRNAs as diagnostic biomarkers and therapeutic targets for the detection of patients at various stages of CRC. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-16T06:51:45.78661-05:0
      DOI: 10.1002/jcp.25801
  • The Cross-Regulation Between SOX15 and Wnt Signaling Pathway
    • Authors: Ali Moradi; Faezeh Ghasemi, Kazem Anvari, Seyed Mahdi Hassanian, Saeideh Ahmadi Simab, Safieh Ebrahimi, Amirreza Hesari, Mohammad Mahdi Forghanifard, Mohammad Taher Boroushaki, Soodabeh ShahidSales, Amir Avan
      Abstract: WNT/B-CATENIN signaling pathway is one of the key dysregulated pathways in different tumor types, which regulate the expression of several genes involved in cell proliferation, differentiation, and survival. This pathway is being modulated by sex-determining region Y-box (SOX) family genes. The functions of these genes are suggested as tumor suppressor or oncogene. SOX genes transcribe a group of transcription factors that play important roles in embryonic development and carcinogenesis. Among them, SOX15 is recently been identified as a novel tumor suppressor in pancreatic and esophagus cancers with a potential role in modulating Wnt/b-catenin signaling. This report summarizes the current knowledge about Wnt/b-catenin signaling pathway and its cross talk with SOX15 with particular emphasis on the value of SOX gene expression as prognostic or predictive biomarker in cancer. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-16T06:51:42.456412-05:
      DOI: 10.1002/jcp.25802
  • Evaluation the Ethnopharmacological Studies in Iran During
           2004–2016: A Systematic Review
    • Authors: Zahra Sadeghi; Maryam Akaberi, Seyed Ahmad Emami, Alireza Sobhkhizi, Amirhossein Sahebkar
      Abstract: Although Iran has a deep history in herbal medicine and great heritage of ancient medical scholars, few efforts have been made to evaluate ethnopharmacological aspects of medicinal plants in this country. In the present study, the authors have reviewed all important literature about the ethnopharmacological investigations on medicinal plants used in the last decade in Iran. All provinces of Iran were categorized according to a phytogeographical division. Information was collected through bibliographic investigations from scientific journals and books. Afterwards, the data were analyzed through the construction of specific ecological regions of the country. Fifty-three references reporting medicinal plants in 5 ecological zones were retrieved. The Irano-Turanian subregion has produced the greatest number of publications in this field among others (47%). Results illustrate that the most reported botanical families were Lamiaceae and Asteraceae (28.57% and 27.73%, respectively). Among various illnesses reported for these plants, gastrointestinal (30.15%), respiratory problems (14.28%), diabetes (11.11%) and cold/flu (11.11%) were the most cited. The most frequently cited medicinal uses were attributed to decoction and infusion preparations. Iran has a rich history of knowledge about phytotherapy and has also a diverse geographical regions and a plant flora that is a good candidate for drug discovery. Documentation of indigenous knowledge about herbal medicine used by Iranian tribes is vital for the future development of herbal drugs. Ethnopharmacological studies of Iranian folk medicine with quantitative analytical techniques are warranted to find drug candidates, and also to preserve the precious knowledge of the Iranian folk medicine. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-16T06:51:35.29241-05:0
      DOI: 10.1002/jcp.25803
  • State of the Art in MicroRNA as Diagnostic and Therapeutic Biomarkers in
           Chronic Lymphocytic Leukemia
    • Authors: Hamed Mirzaei; Sima Fathullahzadeh, Razieh Khanmohammadi, Mansoreh Darijani, Fatemeh Momeni, Aria Masoudifar, Mohammad Goodarzi, Omid Mardanshah, Jan Stanveng, Mahmoud Reza Jaafari, Hamid Reza Mirzaei
      Abstract: Early diagnostic is one of the most important steps in cancer therapy which helps to design and choose a better therapeutic approach. The finding of biomarkers in various levels including genomics, transcriptomics and proteomics levels could provide better treatment for various cancers such as chronic lymphocytic leukemia (CLL). The CLL is the one of main lymphoid malignancies which is specified by aggregation of mature B lymphocytes. Among different biomarkers (e.g. CD38, chromosomes abnormalities, ZAP-70, TP53 and miRNA), microRNAs (miRNAs) have appeared as new diagnostic and therapeutic biomarkers in patients with the CLL disease. Multiple lines of evidence indicated that deregulation of miRNAs could be associated with pathological events which are present in the CLL. These molecules have an effect on a variety of targets such as Bcl2, c-fos, c-Myc, TP53, TCL1 and STAT3 which play critical roles in the CLL pathogenesis. It has been shown that expression of miRNAs could lead to the activation of B cells and BCR. Moreover, exosomes containing miRNAs are one of the other molecules which could contribute to BCR stimulation and progression of CLL cells. Hence, miRNAs and exosomes released from CLL cells could be used as potential diagnostic and therapeutic biomarkers for CLL. This critical review focuses on a very important aspect of CLL based on biomarker discovery covers the pros and cons of using miRNAs as important diagnostics and therapeutics biomarkers for this deadly disease. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-13T06:25:50.025033-05:
      DOI: 10.1002/jcp.25799
  • Gating Modulation of the Tumor-Related Kv10.1 Channel by Mibefradil
    • Authors: Froylán Gómez-Lagunas; Elisa Carrillo, Luis A. Pardo, Walter Stühmer
      Abstract: Several reports credit mibefradil with tumor suppressing properties arising from its known inhibition of Ca2+ currents. Given that mibefradil (Mb) is also known to inhibit K+ channels, we decided to study the interaction between this organic compound and the tumor-related Kv10.1 channel. Here we report that Mb modulates the gating of Kv10.1. Mb induces an apparent inactivation from both open and early closed states where the channels dwell at hyperpolarized potentials. Additionally, Mb accelerates the kinetics of current activation, in a manner that depends on initial conditions. Our observations suggest that Mb binds to the voltage sensor domain of Kv10.1 channels, thereby modifying the gating of the channels in a way that in some, but not all, aspects opposes to the gating effects exerted by divalent cations. J. Cell. Physiol. 9999: 1–14, 2016. © 2016 Wiley Periodicals, Inc.Several reports credit mibefradil, a T-type calcium channel inhibitor, with tumor suppressing properties. Given that mibefradil also inhibits some potassium channels, we studied the interaction of this compound with the tumor related Kv10.1 channel. We found that mibefradil induces an apparent inactivation both from the open and early closed states, and accelerates current activation in a manner that depends on the holding potential. We hypothesize that mibefradil binds to the voltage-sensor module altering the gating of the channels.
      PubDate: 2017-01-11T14:50:36.957889-05:
      DOI: 10.1002/jcp.25448
  • Knockdown of REV3 Synergizes with ATR Inhibition to Promote Apoptosis
           Induced by Cisplatin in Lung Cancer Cells
    • Authors: He-Guo Jiang; Ping Chen, Jin-Yu Su, Ming Wu, Hai Qian, Yi Wang, Jian Li
      Abstract: It has been demonstrated that REV3, the catalytic subunit of the translesion synthesis (TLS) polymerase ζ, play an important role in DNA damage response (DDR) induced by cisplatin, and Ataxia telangietasia mutated and Rad-3-related (ATR) knase is a central player in activating cell cycle checkpoint, stabilizing replication forks, regulating DDR, and promoting repair of DNA damage caused by cisplatin. Cancer cells deficient in either one of REV3 and ATR are more sensitive to cisplatin. However, whether co-inhibition of REV3 and ATR can further increase sensitivity of non-small cell lung cancer (NSCLC) cells to cisplatin is not clear. In this study, we show that REV3 knockdown combined with ATR inhibition further enhance cytotoxicity of cisplatin in NSCLC cells, including cisplatin-sensitive and -resistant cell lines, compared to individual knockdown of REV3 or ATR, which are accompanied by markedly caspase-dependent apoptosis response, pronounced DNA damage accumulation and severe impediment of interstrand crosslink (ICL) and double strand break (DSB) repair. Our results suggest that REV3 knockdown synergize strongly with ATR inhibition to significantly increase sensitivity of cisplatin in NSCLC cells by inhibiting ICL and DSB repair. Thus simultaneously targeting REV3 and ATR may represent one approach to overcome cisplatin resistance and improve chemotherapeutic efficacy in NSCLC treatment. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-11T06:50:38.598198-05:
      DOI: 10.1002/jcp.25792
  • Modulation of Long-Term Endothelial-Barrier Integrity Is Conditional to
           the Cross-Talk between Akt and Src Signaling
    • Authors: Fei Gao; Harika Sabbineni, Sandeep Artham, Payaningal R. Somanath
      Abstract: Although numerous studies have implicated Akt and Src kinases in vascular endothelial growth factor (VEGF) and Angiopoietin-1 (Ang-1)-induced endothelial-barrier regulation, a link between these two pathways has never been demonstrated. We determined the long-term effects of Akt inhibition on Src activity and vice versa, and in turn, on the human microvascular endothelial cell (HMEC) barrier integrity at the basal level, and in response to growth factors. Our data showed that Akt1 gene knockdown increases gap formation in HMEC monolayer at the basal level. Pharmacological inhibition of Akt, but not Src resulted in exacerbated VEGF-induced vascular leakage and impaired Ang-1-induced HMEC-barrier protection in-vitro at 24 hours. Whereas inhibition of Akt had no effect on VEGF-induced HMEC gap formation in the short term, inhibition of Src blunted this process. In contrast, inhibition of Akt disrupted the VEGF and Ang-1 stabilized barrier integrity in the long-term while inhibition of Src did not. Interestingly, both long-term Akt inhibition and Akt1 gene knockdown in HMECs resulted in increased Tyr416 phosphorylation of Src. Treatment of HMECs with transforming growth factor-β1 (TGFβ1) that inhibited Akt Ser473 phosphorylation in the long-term, activated Src through increased Tyr416 phosphorylation and decreased HMEC-barrier resistance. The effect of TGFβ1 on endothelial-barrier breakdown was blunted in Akt1 deficient HMEC monolayers, where endothelial-barrier resistance was already impaired compared to the control. To our knowledge, this is the first report demonstrating a direct cross-talk between Akt and Src in endothelial-barrier regulation. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-11T06:40:34.87694-05:0
      DOI: 10.1002/jcp.25791
  • Current Status and Prospective Regarding the Therapeutic Potential of
           Natural Autoantibodies in Cancer Therapy
    • Authors: Salimeh Ebrahimnezhad; MirHadi Jazayeri, Seyed Mahdi Hassanian, Amir Avan
      Abstract: The presence of natural auto-antibodies (NAbs) in normal range/activity in healthy individuals is essential for body to maintain hemostasis, which are directed to self and altered self-components, while abnormal activity of this system can be associated with several health related diseases. It has been shown that NAbs regulate immune system, and can be changed during the individual's life. In other word, the level and pattern of Nabs is among the main factors to define the state of the body, suggesting their prognostic values as markers of immune system impairment such as autoimmunity and cancer. Such NAbs have gained substantial attention because several of them, including their recombinant forms, have therapeutic potential (e.g., programmed cell death-1 (PD-1, Pdcd1), which some of its inhibitors have been approved by FDA for cancer therapy).Whereas a large number of IgM and IgG NAbs have a key role in tissue homeostasis, while others modulate cellular and enzyme properties. The aim of current review is to give an overview about some of these NAbs and how these low-titer/affinity interact with Ag in homeostasis, with particular emphasis on related diseases such as systemic inflammatory response syndrome and cancer, and their application as potential therapeutic target for cancer therapy. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-07T05:30:22.188017-05:
      DOI: 10.1002/jcp.25765
  • Exosomes: Nanoparticulate Tools for RNA Interference and Drug Delivery
    • Authors: Fahimeh Shahabipour; Nastaran Barati, Thomas P. Johnston, Giuseppe Derosa, Pamela Maffioli, Amirhossein Sahebkar
      Abstract: Exosomes are naturally occurring extracellular vesicles released by most mammalian cells in all body fluids. Exosomes are known as key mediators in cell-cell communication and facilitate the transfer of genetic and biochemical information between distant cells. Structurally, exosomes are composed of lipids, proteins, and also several types of RNAs which enable these vesicles to serve as important disease biomarkers. Moreover, exosomes have emerged as novel drug and gene delivery tools owing to their multiple advantages over conventional delivery systems. Recently, increasing attention has been focused on exosomes for the delivery of drugs, including therapeutic recombinant proteins, to various target tissues. Exosomes are also promising vehicles for the delivery of microRNAs and small interfering RNAs, which is usually hampered by rapid degradation of these RNAs, as well as inefficient tissue specificity of currently available delivery strategies. This review highlights the most recent accomplishments and trends in the use of exosomes for the delivery of drugs and therapeutic RNA molecules. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-07T05:25:40.879746-05:
      DOI: 10.1002/jcp.25766
  • PCSK9 at the Crossroad of Cholesterol Metabolism and Immune Function
           During Infections
    • Authors: Francesco Paciullo; Francesca Fallarino, Vanessa Bianconi, Massimo R. Mannarino, Amirhossein Sahebkar, Matteo Pirro
      Abstract: Sepsis, a complex and dynamic syndrome resulting from microbial invasion and immune system dysregulation, is associated with an increased mortality, reaching up to 35% worldwide. Cholesterol metabolism is often disturbed during sepsis, with low plasma cholesterol levels being associated with poor prognosis. Proprotein convertase subtilisin/kexin type 9 (PCSK9) promotes degradation of the low-density lipoprotein receptor (LDLR), thus regulating intracellular and plasma cholesterol levels. PCSK9 is often upregulated during sepsis and might have a detrimental effect on immune host response and survival. Accordingly, PCSK9 reduces lipopolysaccharide uptake and clearance by human hepatocytes. Moreover, PCSK9 upregulation exacerbates organ dysfunction and tissue inflammation during sepsis, whereas a protective effect of PCSK9 deficiency has been documented in septic patients. Although a possible detrimental impact of PCSK9 on survival has been described, some beneficial effects of PCSK9 on immune response may be hypothesized. First, PCSK9 is associated with increased plasma cholesterol levels, which might be protective during sepsis. Second, PCSK9, by stimulating LDLR degradation and inhibiting reverse cholesterol transport, might promote preferential cholesterol accumulation in macrophages and other immune cells; these events might improve lipid raft composition and augment toll-like receptor function thus supporting inflammatory response. Hence, a more clear definition of the role of PCSK9 in septic states might provide additional insight in the understanding of the sepsis-associated immune dysregulation and enhance therapeutic outcomes. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-07T05:20:28.457716-05:
      DOI: 10.1002/jcp.25767
  • Cyclic Tensile Strain Reduces TNF-α Induced Expression of MMP-13 by
           Condylar Temporomandibular Joint Cells
    • Authors: Hessam Tabeian; Astrid D. Bakker, Beatriz F. Betti, Frank Lobbezoo, Vincent Everts, Teun J. de Vries
      Abstract: To investigate whether the disproportionate degradation of mandibular condyle cartilage in arthritic juvenile temporomandibular joint (TMJ) is related to distinctive responses of TMJ-derived cells to tumor necrosis factor-α (TNF-α), and whether mechanical loading affects this response. The effect of TNF-α (0.1–10 ng/ml) was tested on juvenile porcine TMJ cells isolated from the condyle, fossa, and disc, grown in 3D agarose gels. Expression of anabolic and catabolic factors was quantified by RT-qPCR and/or immunohistochemistry. Condylar cells were stimulated for 12 h with TNF-α (10 ng/ml), followed by 8 h of 6% cyclic tensile strain, and gene expression of MMPs was quantified. TNF-α (10 ng/ml) reduced the expression of the matrix proteins collagen types I and II after 6 h of incubation. Aggrecan gene expression was increased in the presence of 0.1 ng/ml TNF-α. The fossa and disc cells responded to TNF-α with an increased expression of the aggrecanase ADAMTS4. TNF-α enhanced MMP-13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect. Cells isolated from the different cartilaginous structures reacted differently to TNF-α. Since the disc and fossa contain a very low level of proteoglycans in comparison to the condyle, the role played by ADAMTS4 in degradation of the fossa and disc might be limited. TNF-α induced MMP-13 expression by condylar cells might be involved in the degradation of the juvenile condyle. Since this expression was reduced by mechanical loading, functional loading with oral physiotherapy or orthodontic activators may help to reduce the catabolic effect of TNF-α. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.Cells isolated from the different cartilaginous structures reacted differently to TNF-α. TNF-α enhanced MMP-13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect.
      PubDate: 2017-01-06T10:30:23.595916-05:
      DOI: 10.1002/jcp.25593
  • The Emerging Role of TRAF7 in Tumor Development
    • Authors: Tiziana Zotti; Ivan Scudiero, Pasquale Vito, Romania Stilo
      Abstract: The seven members of the tumor necrosis factor receptor (TNF-R)-associated factor (TRAF) family of intracellular proteins were originally discovered and characterized as signaling adaptor molecules coupled to the cytoplasmic regions of receptors of the TNF-R superfamily. Functionally, TRAFs act both as a scaffold and/or enzymatic proteins to regulate activation of mitogen-activated protein kinases (MAPKs) and transcription factors of nuclear factor-κB family (NF-κB). Given the wide variety of stimuli intracellularly conveyed by TRAF proteins, they are physiologically involved in multiple biological processes, including embryonic development, tissue homeostasis, and regulation of innate and adaptive immune responses. In the last few years, it has become increasingly evident the involvement of TRAF7, the last member of the TRAF family to be discovered, in the genesis and progression of several human cancers, placing TRAF7 in the spotlight as a novel tumor suppressor protein. In this paper, we review and discuss the literature recently produced on this subject. J. Cell. Physiol. 9999: 1–6, 2016. © 2016 The
      Authors . Journal of Cellular Physiology Published by Wiley Periodicals, Inc.TRAF proteins are signaling adaptor molecules that regulate activation of mitogen-activated protein kinases (MAPKs) and transcription factors of nuclear factor-κB family (NF-κB). In this paper, we review and discuss the literature recently produced pointing to an involvement of TRAF7 in the genesis and progression of several human cancers.
      PubDate: 2017-01-06T10:15:36.42382-05:0
      DOI: 10.1002/jcp.25676
  • A Novel Skin Splint for Accurately Mapping Dermal Remodeling and
           Epithelialization During Wound Healing
    • Authors: Shiro Jimi; Francesco De Francesco, Giuseppe A. Ferraro, Michele Riccio, Shuuji Hara
      Abstract: The mouse excisional dorsal full-thickness wound model with a silicon splint fixed on the skin has been widely used to mimic human wound healing. However, the method cannot accurately quantify dermal remodeling, since the initial point of epithelialization on the wound surface is unclear. To overcome this limitation, we have developed a novel mouse excisional wound model to assess the degree of epithelial extension and regeneration, using a plastic ring-shaped splint fixed beneath the surrounding epidermal tissue. At the end of the experiment, tissue samples were fixed in formalin, the splint was excised, and paraffin sections were prepared. Splint holes, corresponding to the prior location of the splint, were evident on the tissue cross-sections, and the epidermis above the holes was considered the initial excision site. The epidermal contraction and epithelial regeneration, as independent essential tissue alterations in wound healing, could be distinguishable and quantified. Compared with previous splint models, this method provides an accurate evaluation of epidermal processes in wound healing, and can be a platform to assess the effects of various wound healing factors. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.The new method established in this manuscript makes possible the accurate assessment and measurement of epidermal changes—dermal remodeling and regeneration—in the wound healing process in mice.
      PubDate: 2017-01-06T10:15:32.764507-05:
      DOI: 10.1002/jcp.25595
  • Roles of Multifunctional COP9 Signalosome Complex in Cell Fate and
           Implications for Drug Discovery
    • Authors: Ping Li; Longxiang Xie, Yinzhong Gu, Jiang Li, Jianping Xie
      Abstract: The eight subunits containing COP9 signalosome (CSN) complex, is highly conserved among eukaryotes. CSN, identified as a negative regulator of photomorphogenesis, has also been demonstrated to be important in proteolysis, cellular signal transduction and cell cycle regulation in various eukaryotic organisms. This review mainly summarizes the roles of CSN in cell cycle regulation, signal transduction and apoptosis, and its potential as diagnostic biomarkers, drug targets for cancer and infectious diseases. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.This review mainly summarizes the roles of CSN in cell cycle regulation, signal transduction and apoptosis, and its potential as diagnostic biomarkers, drug targets for cancer and infectious diseases.
      PubDate: 2017-01-05T08:45:23.116809-05:
      DOI: 10.1002/jcp.25696
  • A MicroRNA Cluster miR-23–24–27 Is Upregulated by Aldosterone in the
           Distal Kidney Nephron Where it Alters Sodium Transport
    • Authors: Xiaoning Liu; Robert S. Edinger, Christine A. Klemens, Yu L. Phua, Andrew J. Bodnar, William A. LaFramboise, Jacqueline Ho, Michael B. Butterworth
      Abstract: The epithelial sodium channel (ENaC) is expressed in the epithelial cells of the distal convoluted tubules, connecting tubules, and cortical collecting duct (CCD) in the kidney nephron. Under the regulation of the steroid hormone aldosterone, ENaC is a major determinant of sodium (Na+) and water balance. The ability of aldosterone to regulate microRNAs (miRs) in the kidney has recently been realized, but the role of miRs in Na+ regulation has not been well established. Here we demonstrate that expression of a miR cluster mmu-miR-23–24–27, is upregulated in the CCD by aldosterone stimulation both in vitro and in vivo. Increasing the expression of these miRs increased Na+ transport in the absence of aldosterone stimulation. Potential miR targets were evaluated and miR-27a/b was verified to bind to the 3′-untranslated region of intersectin-2, a multi-domain protein expressed in the distal kidney nephron and involved in the regulation of membrane trafficking. Expression of Itsn2 mRNA and protein was decreased after aldosterone stimulation. Depletion of Itsn2 expression, mimicking aldosterone regulation, increased ENaC-mediated Na+ transport, while Itsn2 overexpression reduced ENaC's function. These findings reinforce a role for miRs in aldosterone regulation of Na+ transport, and implicate miR-27 in aldosterone's action via a novel target. J. Cell. Physiol. 9999: 1–12, 2016. © 2016 Wiley Periodicals, Inc.The steroid hormone aldosterone alters the expression of microRNAs in distal kidney nephron epithelial cells, and coordinately upregulates the miR-23–24–27 cluster. MiR-27 targets intersectin 2 to reduce its expression. Decreased intersectin 2 levels results in an increase in the activity of the epithelial sodium channel to augment sodium transport.
      PubDate: 2017-01-05T08:35:27.33984-05:0
      DOI: 10.1002/jcp.25599
  • Identifying Nuclear Matrix-Attached DNA Across the Genome
    • Authors: Jason R. Dobson; Deli Hong, A. Rasim Barutcu, Hai Wu, Anthony N. Imbalzano, Jane B. Lian, Janet L. Stein, Andre J. van Wijnen, Jeffrey A. Nickerson, Gary S. Stein
      Abstract: Experimental approaches to define the relationship between gene expression and nuclear matrix attachment regions (MARs) have given contrasting and method-specific results. We have developed a next generation sequencing strategy to identify MARs across the human genome (MAR-Seq). The method is based on crosslinking chromatin to its nuclear matrix attachment sites to minimize changes during biochemical processing. We used this method to compare nuclear matrix organization in MCF-10A mammary epithelial-like cells and MDA-MB-231 breast cancer cells and evaluated the results in the context of global gene expression (array analysis) and positional enrichment of gene-regulatory histone modifications (ChIP-Seq). In the normal-like cells, nuclear matrix-attached DNA was enriched in expressed genes, while in the breast cancer cells, it was enriched in non-expressed genes. In both cell lines, the chromatin modifications that mark transcriptional activation or repression were appropriately associated with gene expression. Using this new MAR-Seq approach, we provide the first genome-wide characterization of nuclear matrix attachment in mammalian cells and reveal that the nuclear matrix-associated genome is highly cell-context dependent. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.MAR-Seq is a next-generation sequencing strategy to identify matrix attachment regions (MARs) that is based on crosslinking chromatin to its nuclear attachment sites. Comparison of MCF-10A mammary epithelial-like cells and MDA-MB-231 breast cancer cells showed that nuclear matrix-attached DNA in normal-like MCF-10A cells is enriched in expressed genes, while in the breast cancer cells, it is enriched in non-expressed genes. This new MAR-Seq approach reveals that the nuclear matrix-associated genome is highly cell-context dependent.
      PubDate: 2017-01-05T08:35:24.082065-05:
      DOI: 10.1002/jcp.25596
  • Fibroblast Growth Factor 21 (FGF21) Promotes Formation of Aerobic
           Myofibers via the FGF21-SIRT1-AMPK-PGC1α Pathway
    • Authors: Xinyi Liu; Yongliang Wang, Liming Hou, Yuanzhu Xiong, Shuhong Zhao
      Abstract: Fibroblast growth factor 21(FGF21) is a pivotal regulator of energy metabolism, which is currently being assessed as a potential drug target for the treatment of insulin-resistant conditions. However, the cellular mechanisms by which FGF21 affects myogenesis remain unclear. In this study, we explored the function of FGF21 in myogenesis both in vitro and in vivo. Our experiments showed for the first time that FGF21 promotes myoblast differentiation and serves as a switch of molecular transformation from anaerobic myofibers to aerobic myofibers via the FGF21-SIRT1-AMPK-PGC1α axis. Furthermore, we employed the Dual-Luciferase Reporter Assay System and Electrophoretic Mobility Shift Assay (EMSA) and demonstrated that MYOD, a major myogenic transcription factor, binds directly to the promoter region of Fgf21, leading to the activation of Fgf21 expression in mouse C2C12 myoblasts. Our study revealed a novel mechanism of myogenesis and muscle fiber transformation and indicated that FGF21 serves as a vital regulator of muscle development and important contributor to the pathogenesis of myopathy. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-05T08:30:37.504911-05:
      DOI: 10.1002/jcp.25735
  • Calretinin Immunoreactivity in the Human Testis Throughout Fetal Life
    • Authors: Giovanna G. Altobelli; Francesca Pentimalli, Mariarosaria D'Armiento, Susan Van Noorden, Vincenzo Cimini
      Abstract: The main functions of the testis are sex hormone and sperm cell production. Steroidogenesis occurs in the Leydig interstitial cells and spermatogenesis in the seminiferous tubules. Male gonad morphogenesis is a finely orchestrated process, mainly coordinated by hormones, whose actions can significantly affect post-pubertal testicular function. Calcium is a key intracellular messenger, which regulates many signal transduction pathways, and is also implicated in steroidogenesis. Calcium homeostasis and signaling rely on many calcium-binding proteins including calretinin, of the "EF-hand" protein family. Calretinin is a highly conserved protein mainly expressed in the nervous system but also detected in rat and human adult and fetal testis as well as in pathological conditions. Calretinin expression in the fetal testis, however, has not been thoroughly analyzed probably owing to limited availability and paucity of tissues. Here we examined by immunocytochemistry the expression of calretinin in human fetal testis specimens, obtained from natural and therapeutic abortions, at various developmental ages. We found that calretinin-immunoreactive Leydig cells were visible throughout the timeframe studied (14th to 27th week). Immunoreactivity was also observed in Sertoli cells and in the germ cells of the immature seminiferous tubules. Overall our data indicate that calretinin expression parallels the decline in Leydig cell number, suggesting that its presence is indeed correlated to their steroidogenic activity. They also suggest that the intratubular positivity of calretinin could be linked to the ability of Sertoli cells to produce locally acting hormones contributing to the histodifferentiation of the male genital tract. This article is protected by copyright. All rights reserved
      PubDate: 2017-01-05T08:25:28.973764-05:
      DOI: 10.1002/jcp.25727
  • Early Subchondral Bone Loss at Arthritis Onset Predicted Late Arthritis
           Severity in a Rat Arthritis Model
    • Authors: Guillaume Courbon; Damien Cleret, Marie-Thérèse Linossier, Laurence Vico, Hubert Marotte
      Abstract: Synovitis is usually observed before loss of articular function in rheumatoid arthritis (RA). In addition to the synovium and according to the “Inside–Outside” theory, bone compartment is also involved in RA pathogenesis. Then, we investigated time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant-induced arthritis (AIA) model. Lewis female rats were longitudinally monitored from arthritis induction (day 0), with early (day 10) and late (day 17) steps. Trabecular and cortical microarchitecture parameters of four ankle bones were assessed by microcomputed tomography. Gene expression was determined at sacrifice. Arthritis occurred at day 10 in AIA rats. At this time, bone erosions were detected on four ankle bones, with cortical porosity increase (+67%) and trabecular alterations including bone volume fraction (BV/TV: −13%), and trabecular thickness decrease. Navicular bone assessment was the most reproducible and sensitive. Furthermore, strong correlations were observed between bone alterations at day 10 and arthritis severity or bone loss at day 17, including predictability of day 10 BV/TV to day 17 articular index (R2 = 0.76). Finally, gene expression at day 17 confirmed massive osteoclast activation and interestingly provided insights on strong activation of bone formation inhibitor markers at the joint level. In rat AIA, bone loss was already observed at synovitis onset and was predicted late arthritis severity. Our results reinforced the key role of subchondral bone in arthritis pathogenesis, in favour to the “Inside–Outside” theory. Mechanisms of bone loss in rat AIA involved resorption activation and formation inhibition changes. J. Cell. Physiol. 9999: 1–8, 2016. © 2016 Wiley Periodicals, Inc.We explored time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant-induced arthritis (AIA) model. At clinical synovitis onset, bone porosity and microarchitecture alterations were recorded in the ankle bones. These early modifications were predictive of arthritis outcome, underlining the importance of early bone loss investigation.
      PubDate: 2017-01-05T08:15:26.639024-05:
      DOI: 10.1002/jcp.25601
  • Table of Contents, Editor's Choice, Highlights
    • Pages: 889 - 895
      PubDate: 2017-01-19T10:00:53.178496-05:
      DOI: 10.1002/jcp.25757
  • Primary Cilium-Regulated EG-VEGF Signaling Facilitates Trophoblast
    • Authors: Chia-Yih Wang; Hui-Ling Tsai, Jhih-Siang Syu, Ting-Yu Chen, Mei-Tsz Su
      Abstract: Trophoblast invasion is an important event in embryo implantation and placental development. During these processes, endocrine gland-derived vascular endothelial growth factor (EG-VEGF) is the key regulator mediating the crosstalk at the feto-maternal interface. The primary cilium is a cellular antenna receiving environmental signals and is crucial for proper development. However, little is known regarding the role of the primary cilium in early human pregnancy. Here, we demonstrate that EG-VEGF regulates trophoblast cell invasion via primary cilia. We found that EG-VEGF activated ERK1/2 signaling and subsequent upregulation of MMP2 and MMP9, thereby facilitating cell invasion in human trophoblast HTR-8/SVneo cells. Inhibition of ERK1/2 alleviated the expression of MMPs and trophoblast cell invasion after EG-VEGF treatment. In addition, primary cilia were observed in all the trophoblast cell lines tested and, more importantly, in human first-trimester placental tissue. The receptor of EG-VEGF, PROKR1, was detected in primary cilia. Depletion of IFT88, the intraflagellar transporter required for ciliogenesis, inhibited primary cilium growth, thereby ameliorating ERK1/2 activation, MMP upregulation, and trophoblast cell invasion promoted by EG-VEGF. These findings demonstrate a novel function of primary cilia in controlling EG-VEGF-regulated trophoblast invasion and reveal the underlying molecular mechanism. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.Primary cilium is a cellular antenna that conveys extracellular signals into a cellular response. EG-VEGF binds to PROKR1 locating on the cilium, and activates ERK1/2 signaling followed by upregulating MMP2 and MMP9 for facilitating cell invasion. We demonstrated that EG-VEGF regulated trophoblast cell invasion via primary cilium in the present study.
      PubDate: 2016-12-29T05:40:59.355062-05:
      DOI: 10.1002/jcp.25649
  • Pancreatic Endoderm-Derived From Diabetic Patient-Specific Induced
           Pluripotent Stem Cell Generates Glucose-Responsive Insulin-Secreting Cells
    • Authors: Bahareh Rajaei; Mehdi Shamsara, Masoumeh Fakhr Taha, Leila Mohammadi Amirabad, Mohammad Massumi, Mohammad Hossein Sanati
      Abstract: Human-induced pluripotent stem cells (hiPSCs) can potentially serve as an invaluable source for cell replacement therapy and allow the creation of patient- and disease-specific stem cells without the controversial use of embryos and avoids any immunological incompatibility. The generation of insulin-producing pancreatic β-cells from pluripotent stem cells in vitro provides an unprecedented cell source for personal drug discovery and cell transplantation therapy in diabetes. A new five-step protocol was introduced in this study, effectively induced hiPSCs to differentiate into glucose-responsive insulin-producing cells. This process mimics in vivo pancreatic organogenesis by directing cells through stages resembling definitive endoderm, primitive gut-tube endoderm, posterior foregut, pancreatic endoderm, and endocrine precursor. Each stage of differentiation were characterized by stage-specific markers. The produced cells exhibited many properties of functional β-cells, including expression of critical β-cells transcription factors, the potency to secrete C-peptide in response to high levels of glucose and the presence of mature endocrine secretory granules. This high efficient differentiation protocol, established in this study, yielded 79.18% insulin-secreting cells which were responsive to glucose five times higher than the basal level. These hiPSCs-derived glucose-responsive insulin-secreting cells might provide a promising approach for the treatment of type I diabetes mellitus. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.A new five-step protocol was introduced in this study, effectively induced hiPSCs to differentiate into glucose-responsive insulin-producing cells. This high efficient differentiation protocol, yielded 79.18% insulin- secreting cells, which were responsive to glucose five times higher than the basal level. These hiPSCs-derived glucose-responsive insulin- secreting cells might provide a promising approach for the treatment of type I diabetes mellitus.
      PubDate: 2016-12-29T05:37:08.821584-05:
      DOI: 10.1002/jcp.25459
  • Exploring the Role of PGC-1α in Defining Nuclear Organisation in
           Skeletal Muscle Fibres
    • Authors: Jacob Alexander Ross; Adam Pearson, Yotam Levy, Bettina Cardel, Christoph Handschin, Julien Ochala
      Abstract: Muscle fibres are multinucleated cells, with each nucleus controlling the protein synthesis in a finite volume of cytoplasm termed the myonuclear domain (MND). What determines MND size remains unclear. In the present study, we aimed to test the hypothesis that the level of expression of the transcriptional coactivator PGC-1α and subsequent activation of the mitochondrial biogenesis are major contributors. Hence, we used two transgenic mouse models with varying expression of PGC-1α in skeletal muscles. We isolated myofibres from the fast twitch extensor digitorum longus (EDL) and slow twitch diaphragm muscles. We then membrane-permeabilised them and analysed the 3D spatial arrangements of myonuclei. In EDL muscles, when PGC-1α is over-expressed, MND volume decreases; whereas, when PGC-1α is lacking, no change occurs. In the diaphragm, no clear difference was noted. This indicates that PGC-1α and the related mitochondrial biogenesis programme are determinants of MND size. PGC-1α may facilitate the addition of new myonuclei in order to reach MND volumes that can support an increased mitochondrial density. J. Cell. Physiol. 9999: 1–5, 2016. © 2016 Wiley Periodicals, Inc.Muscle fibres are multinucleated cells, with individual nuclei controlling the protein synthesis in a finite volume of cytoplasm termed myonuclear domain. The present study indicates that one important regulator of myonuclear domain size is PGC-1α and related mitochondrial biogenesis programme.
      PubDate: 2016-12-29T05:37:05.80091-05:0
      DOI: 10.1002/jcp.25678
  • Lgr4 Expression in Osteoblastic Cells Is Suppressed by Hydrogen Peroxide
    • Authors: Chantida Pawaputanon Na Mahasarakham; Yayoi Izu, Katsuhiko Nishimori, Yuichi Izumi, Masaki Noda, Yoichi Ezura
      Abstract: LGR4 is expressed in bone and has been shown to be involved in bone metabolism. Oxidative stress is one of the key issues in pathophysiology of osteoporosis. However, the link between Lgr4 and oxidative stress has not been known. Therefore, effects of hydrogen peroxide on Lgr4 expression in osteoblasts were examined. Hydrogen peroxide treatment suppressed the levels of Lgr4 mRNA expression in an osteoblastic cell line, MC3T3-E1. The suppressive effects were not obvious at 0.1 mM, while 1 mM hydrogen peroxide suppressed Lgr4 expression by more than 50%. Hydrogen peroxide treatment suppressed Lgr4 expression within 12 h and this suppression lasted at least up to 48 h. Hydrogen peroxide suppression of Lgr4 expression was still observed in the presence of a transcription inhibitor but was no longer observed in the presence of a protein synthesis inhibitor. Although Lgr4 expression in osteoblasts is enhanced by BMP2 treatment as reported before, hydrogen peroxide treatment suppressed Lgr4 even in the presence of BMP2. Finally, hydrogen peroxide suppressed Lgr4 expression in primary cultures of osteoblasts similarly to MC3T3-E1 cells. These date indicate that hydrogen peroxide suppresses Lgr4 expression in osteoblastic cells. J. Cell. Physiol. 9999: 1–6, 2016. © 2016 Wiley Periodicals, Inc.
      PubDate: 2016-12-29T05:37:03.982426-05:
      DOI: 10.1002/jcp.25684
  • Identification of a Novel Human E-Cadherin Splice Variant and Assessment
           of Its Effects Upon EMT-Related Events
    • Authors: María Laura Matos; Lara Lapyckyj, Marina Rosso, María José Besso, María Victoria Mencucci, Clara Isabel Marín Briggiler, Silvina Giustina, Laura Inés Furlong, Mónica Hebe Vazquez-Levin
      Abstract: Epithelial Cadherin (E-cadherin) is involved in calcium-dependent cell–cell adhesion and signal transduction. The E-cadherin decrease/loss is a hallmark of Epithelial to Mesenchymal Transition (EMT), a key event in tumor progression. The underlying molecular mechanisms that trigger E-cadherin loss and consequent EMT have not been completely elucidated. This study reports the identification of a novel human E-cadherin variant mRNA produced by alternative splicing. A bioinformatics evaluation of the novel mRNA sequence and biochemical verifications suggest its regulation by Nonsense-Mediated mRNA Decay (NMD). The novel E-cadherin variant was detected in 29/42 (69%) human tumor cell lines, expressed at variable levels (E-cadherin variant expression relative to the wild type mRNA = 0.05–11.6%). Stable transfection of the novel E-cadherin variant in MCF-7 cells (MCF7Ecadvar) resulted in downregulation of wild type E-cadherin expression (transcript/protein) and EMT-related changes, among them acquisition of a fibroblastic-like cell phenotype, increased expression of Twist, Snail, Zeb1, and Slug transcriptional repressors and decreased expression of ESRP1 and ESRP2 RNA binding proteins. Moreover, loss of cytokeratins and gain of vimentin, N-cadherin and Dysadherin/FXYD5 proteins was observed. Dramatic changes in cell behavior were found in MCF7Ecadvar, as judged by the decreased cell–cell adhesion (Hanging-drop assay), increased cell motility (Wound Healing) and increased cell migration (Transwell) and invasion (Transwell w/Matrigel). Some changes were found in MCF-7 cells incubated with culture medium supplemented with conditioned medium from HEK-293 cells transfected with the E-cadherin variant mRNA. Further characterization of the novel E-cadherin variant will help understanding the molecular basis of tumor progression and improve cancer diagnosis. J. Cell. Physiol. 9999: 1–19, 2016. © 2016 Wiley Periodicals, Inc.This report describes the identification and partial characterization of a novel human E-cadherin variant mRNA produced by alternative splicing. Stable transfection of the E-cadherin variant mRNA in MCF-7 cells (MCF7Ecadvar cells) resulted in downregulation of wild type E-cadherin and EMT-related changes, that is, acquisition of a fibroblast-like cellular phenotype, increased expression of E-cadherin transcriptional repressors, vimentin and N-cadherin, and loss of cytokeratins. MCF7Ecadvar cells depicted dramatic changes in cellular behavior, showing reduced cell–cell adhesion and increased cell migration and invasiveness.
      PubDate: 2016-12-29T05:36:52.820603-05:
      DOI: 10.1002/jcp.25622
  • Differential Expression of miR-4520a Associated With Pyrin Mutations in
           Familial Mediterranean Fever (FMF)
    • Authors: Helen Latsoudis; Mir-Farzin Mashreghi, Joachim R. Grün, Hyun-Dong Chang, Bruno Stuhlmüller, Argyro Repa, Irini Gergiannaki, Eleni Kabouraki, George S. Vlachos, Thomas Häupl, Andreas Radbruch, Prodromos Sidiropoulos, Kimon Doukoumetzidis, Dimitris Kardassis, Timothy B. Niewold, Dimitrios T. Boumpas, George N. Goulielmos
      Abstract: Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute, and self-limiting attacks of fever. Mutations in MEFV gene encoding pyrin account for FMF, but the high number of heterozygote patients with typical symptoms of the disease has driven a number of alternative aetiopathogenic hypotheses. The MEFV gene was knocked down in human myelomonocytic cells that express endogenous pyrin to identify deregulated microRNAs (miRNAs). Microarray analyses revealed 29 significantly differentially expressed miRNAs implicated in pathways associated with cellular integrity and survival. Implementation of in silico gene network prediction algorithms and bioinformatics analyses showed that miR-4520a is predicted to target genes implicated in autophagy through regulation of RHEB/mTOR signaling. Differential expression levels of RHEB were confirmed by luciferase reporter gene assays providing further evidence that is directly targeted by miR-4520a. Although the relative expression levels of miR-4520a were variable among FMF patients, the statistical expression of miR-4520a was different between FMF mutation carriers and controls (P = 0.0061), indicating an association between miR-4520a expression and MEFV mutations. Comparison between FMF patients bearing the M694V mutation, associated with severe disease, and healthy controls showed a significant increase in miR-4520a expression levels (P = 0.00545). These data suggest that RHEB, the main activator of mTOR signaling, is a valid target of miR-4520a with the relative expression levels of the latter being significantly deregulated in FMF patients and highly dependent on the presence of pyrin mutations, especially of the M694V type. These results suggest a role of deregulated autophagy in the pathogenesis of FMF. J. Cell. Physiol. 9999: 1–11, 2016. © 2016 Wiley Periodicals, Inc.Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute and self-limiting attacks of fever. In the present study we presented for the first time results supporting a role of deregulated autophagy in the pathogenesis of FMF.
      PubDate: 2016-12-20T08:30:57.563042-05:
      DOI: 10.1002/jcp.25602
  • A Human 3D In Vitro Model to Assess the Relationship between Osteoporosis
           and Dissemination to Bone of Breast Cancer Tumour Cells
    • Authors: F Salamanna; V Borsari, S Brogini, P Torricelli, S Cepollaro, M Cadossi, M Fini
      Abstract: Despite consistent improvements in diagnostic and therapeutic strategies for breast cancer, up to 40% of patients will develop bone metastases. To reduce the morbidity and complications related with bone metastases, it is imperative to reduce their etiological factors. Osteoporosis, being characterized by a sudden estrogen deficiency, may provide a favorable condition for bone metastasis. This work, using a humanized 3D in vitro model, aims at evaluating the relationship between osteoporosis and breast cancer-derived bone metastases. Bone tissue discarded from total hip replacement surgery of healthy and osteoporotic patients was cultured in a rolling apparatus system in hypoxic environment. Protein levels (i.e. vascular endothelial growth factor (VEGF), VEGF receptor 1, VEGF receptor 2, interleukin (IL)-6, IL-1β, IL-8 IL-10, tumor necrosis factor α (TNF-α), osteoprotegerin (OPG), Receptor Activator for Nuclear factor KB Ligand (RANKL)) and histological and immunohistochemical (i.e. cytokeratin 8 and 18) analyses showed a noticeable specificity of breast cancer cells for the colonization of osteoporotic bone. These data are the first to demonstrate that using humanized 3D in vitro systems, which individually model the pre- and postmenopausal bone microenvironment, it is possible to recognize major differences in tumor growth and colonization between healthy and osteoporotic status. Thus, this system might help to develop a shared system between basic and clinical sciences where a personalized diagnosis is associated to a therapeutic strategy designed for a single patient: a model able to achieve a translational research approach in the clinical setting, which may lead to the application and dissemination of personalized medicine. This article is protected by copyright. All rights reserved
      PubDate: 2016-12-07T07:52:39.43569-05:0
      DOI: 10.1002/jcp.25708
  • SV40 Infection of Mesenchymal Stromal Cells from Wharton's Jelly Drives
           the Production of Inflammatory and Tumoral Mediators
    • Authors: Carolina Cason; Giuseppina Campisciano, Nunzia Zanotta, Erica Valencic, Serena Delbue, Ramona Bella, Manola Comar
      Abstract: The Mesenchymal Stromal Cells from umbilical cord Wharton's jelly (WJSCs) are a source of cells with high potentiality for the treatment of human immunological disorders. Footprints of the oncogenic viruses Simian Virus 40 (SV40) and JC Virus (JCPyV) have been recently detected in human WJSCs specimens. The aim of this study is to evaluate if WJSCs can be efficiently infected by these Polyomaviruses and if they can potentially exert tumoral activity. Cell culture experiments indicated that WJSCs could sustain both SV40 and JCPyV infections. A transient and lytic replication was observed for JCPyV, while SV40 persistently infected WJSCs over a long period of time, releasing a viral progeny at low titer without evident cytopathic effect (CPE). Considering the association between SV40 and human tumors and the reported ability of the oncogenic viruses to drive the host innate immune response to cell transformation, the expression profile of a large panel of immune mediators was evaluated in supernatants by the Bioplex platform. RANTES, IL-3, MIG and IL-12p40, involved in chronic inflammation, cells differentiation and transformation, were constantly measured at high concentration comparing to control. These findings represent a new aspect of SV40 biological activity in the humans, highlighting its interaction with specific host cellular pathways. In view of these results, it seems to be increasingly urgent to consider Polyomaviruses in the management of WJSCs for their safely use as promising therapeutic source. This article is protected by copyright. All rights reserved
      PubDate: 2016-12-07T07:50:49.611089-05:
      DOI: 10.1002/jcp.25723
  • Calprotectin Induces IL-6 and MCP-1 Production via Toll-Like Receptor 4
           Signaling in Human Gingival Fibroblasts
    • Authors: Yasufumi Nishikawa; Yukari Kajiura, Jung Hwan Lew, Jun-ichi Kido, Toshihiko Nagata, Koji Naruishi
      Abstract: Calprotectin, a heterodimer of S100A8 and S100A9 molecules, is associated with inflammatory diseases such as inflammatory bowel disease. We have reported that calprotectin levels in gingival crevicular fluids of periodontitis patients are significantly higher than in healthy subjects. However, the functions of calprotectin in pathophysiology of periodontitis are still unknown. The aim of this study is to investigate the effects of calprotectin on the productivity of inflammatory cytokines in human gingival fibroblasts (HGFs). The HGFs cell line CRL-2014® (ATCC) were cultured, and total RNAs were collected to examine the expression of TLR2/4 and RAGE mRNA using RT-PCR. After the cells were treated with S100A8, S100A9 and calprotectin, supernatants were collected and the levels of IL-6 and MCP-1 were measured using ELISA methods. To examine the intracellular signals involved in calprotectin-induced cytokine production, several chemical inhibitors were used. Furthermore, after the siRNA-mediated TLR4 down-regulated cells were treated with S100A8, S100A9 and calprotectin, the levels of IL-6 and MCP-1 were also measured. HGFs showed greater expression of TLR4 mRNA, but not TLR2 and RAGE mRNA compared with human oral epithelial cells. Calprotectin increased significantly the production of MCP-1 and IL-6 in HGFs, and the cytokine productions were significantly suppressed in the cells treated with MAPKs, NF-κB and TLR4 inhibitors. Furthermore, calprotectin-mediated MCP-1 and IL-6 production were significantly suppressed in TLR4 down-regulated cells. Taken together, calprotectin induces IL-6 and MCP-1 production in HGFs via TLR4 signaling that involves MAPK and NF-κB, resulting in the progression of periodontitis. This article is protected by copyright. All rights reserved
      PubDate: 2016-12-07T07:11:32.759639-05:
      DOI: 10.1002/jcp.25724
  • SIRT1-SIRT3 Axis Regulates Cellular Response to Oxidative Stress and
    • Authors: Ilaria Carnevale; Laura Pellegrini, Patrizia D'Aquila, Serena Saladini, Emanuela Lococo, Lucia Polletta, Enza Vernucci, Eleonora Foglio, Stefano Coppola, Luigi Sansone, Giuseppe Passarino, Dina Bellizzi, Matteo A. Russo, Massimo Fini, Marco Tafani
      Abstract: Sirtuins are conserved NAD+-dependent deacylases. SIRT1 is a nuclear and cytoplasmic sirtuin involved in the control of histones a transcription factors function. SIRT3 is a mitochondrial protein, which regulates mitochondrial function. Although both SIRT1 and SIRT3 have been implicated in resistance to cellular stress, the link between these two sirtuins has not been studied so far. Here we aimed to unravel: i) the role of SIRT1-SIRT3 axis for cellular response to oxidative stress and DNA damage; ii) how mammalian cells modulate such SIRT1-SIRT3 axis and which mechanisms are involved. Therefore, we analyzed the response to different stress stimuli in WT or SIRT1-silenced cell lines. Our results demonstrate that SIRT1-silenced cells are more resistant to H2O2 and etoposide treatment showing decreased ROS accumulation, γ-H2AX phosphorylation, caspase-3 activation and PARP cleavage. Interestingly, we observed that SIRT1-silenced cells show an increased SIRT3 expression. To explore such a connection, we carried out luciferase assays on SIRT3 promoter demonstrating that SIRT1-silencing increases SIRT3 promoter activity and that such an effect depends on the presence of SP1 and ZF5 recognition sequences on SIRT3 promoter. Afterwards, we performed co-immunoprecipitation assays demonstrating that SIRT1 binds and deacetylates the transcription inhibitor ZF5 and that there is a decreased interaction between SP1 and ZF5 in SIRT1-silenced cells. Therefore, we speculate that acetylated ZF5 cannot bind and sequester SP1 that is free, then, to increase SIRT3 transcription.In conclusion, we demonstrate that cells with low SIRT1 levels can maintain their resistance and survival by increasing SIRT3 expression. This article is protected by copyright. All rights reserved
      PubDate: 2016-12-07T07:08:05.755569-05:
      DOI: 10.1002/jcp.25711
  • Condensin Regulation of Genome Architecture
    • Authors: Vibhuti Rana; Giovanni Bosco
      Abstract: Progression through the cell cycle for both prokaryotes and eukaryotes is required for cell division. At almost every level of cellular organization structure and function of subcellular compartments and organelles must accommodate the progression of cell division. As cells progress through the cell division cycle, perhaps the most dramatic observable cellular change is the shape of chromosomes. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-26T01:35:40.940435-05:
      DOI: 10.1002/jcp.25702
  • Modulation of Chloride Currents in Human Lung Epithelial Cells Exposed to
           Exogenous Oxidative Stress
    • Authors: Rita Canella; Marta Martini, Roberta Borriello, Carlotta Cavicchio, Ximena M Muresan, Mascia Benedusi, Franco Cervellati, Giuseppe Valacchi
      Abstract: Air pollution continues to be a major public health concern affecting 9 out of 10 individuals living in urban areas worldwide. Respiratory tract is the organ most exposed to gas pollution and ozone has been shown to be one of the most noxious pollutants to which living organisms are exposed. In the present work, we have investigated the effects of 0.1 ppm of ozone on chloride currents in human lung epithelial cells (A549 line) and whether this effect could be modulated by vitamin E pre-treatment.Whole-cell patch clamp technique was applied to not excitable cells in order to obtain information about chloride currents behavior, important for epithelial lung cells homeostasis. Significant alteration of the I-V curve after ozone treatment was observed, with the appearance of a large outward rectifier component decreasing over time and returning to the basal state levels after 24 hours. Statistical analysis indicated a modification of the amount of ions passing the membrane in the unit of time as a possible cause of this difference. RT-qPCR analysis showed an increase in ClC-2 and ORCC mRNA after ozone exposure.In addition, pre-treatment with vitamin E was able to suppress the outward rectifier component induced by ozone, bringing back the current values to the control level and preventing ozone induced chloride channels up regulation.Our data suggest that ozone exposure is able to modify chloride current density and the use of vitamin E can prevent the above mentioned damage. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-25T08:05:22.13479-05:0
      DOI: 10.1002/jcp.25705
  • Roles of Notch Signaling in Adipocyte Progenitor Cells and Mature
    • Authors: Tizhong Shan; Jiaqi Liu, Weiche Wu, Ziye Xu, Yizhen Wang
      Abstract: Adipose tissues, composed with mature adipocytes and preadipocytic stromal/stem cells, play crucial roles in whole body energy metabolism and regenerative medicine. Mature adipocytes are derived and differentiated from mesenchymal stem cells (MSCs) or preadipocytes. This differentiation process, also called adipogenesis, is regulated by several signaling pathways and transcription factors. Notch1 signaling is a highly conserved pathway that is indispensable for stem cell hemostasis and tissue development. In adipocyte progenitor cells, Notch1 signaling regulates the adipogenesis process including proliferation and differentiation of the adipocyte progenitor cells in vitro. Notably, the roles of Notch1 signaling in beige adipocytes formation, adipose development and function, and the whole body energy metabolism have been recently reported. Here, we mainly review and discuss the roles of Notch1 signaling in adipogenesis in vitro as well as in beige adipocytes formation, adipocytes dedifferentiation and function in vivo. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-21T07:10:20.732207-05:
      DOI: 10.1002/jcp.25697
  • Calcium Signaling During Meiotic Cell Cycle Regulation and Apoptosis in
           Mammalian Oocytes
    • Authors: Meenakshi Tiwari; Shilpa Prasad, Tulsidas G. Shrivastav, Shail K. Chaube
      Abstract: Calcium (Ca++) is one of the major signal molecules that regulate various aspects of cell functions including cell cycle progression, arrest and apoptosis in wide variety of cells. This review summarizes current knowledge on the differential roles of Ca++ in meiotic cell cycle resumption, arrest and apoptosis in mammalian oocytes. Release of Ca++ from internal stores and/or Ca++ influx from extracellular medium causes moderate increase of intracellular Ca++ ([Ca++]i) level and reactive oxygen species (ROS). Increase of Ca++ as well as ROS levels under physiological range trigger maturation promoting factor (MPF) destabilization thereby meiotic resumption from diplotene as well as metaphase-II (M-II) arrest in oocytes. A sustained increase of [Ca++]i level beyond physiological range induces generation of ROS sufficient enough to cause oxidative stress (OS) in aging oocytes. The increased [Ca++]i triggers Fas ligand-mediated oocyte apoptosis. Further, OS triggers mitochondria-mediated oocyte apoptosis in several mammalian species. Thus, Ca++ exerts differential roles on oocyte physiology depending upon its intracellular concentration. A moderate increase of [Ca++]i as well as ROS mediate spontaneous resumption of meiosis from diplotene as well as M-II arrest, while their high levels cause meiotic cell cycle arrest and apoptosis by operating both mitochondria- as well as Fas ligand-mediated apoptotic pathways. Indeed, Ca++ regulates cellular physiology by modulating meiotic cell cycle and apoptosis in mammalian oocytes. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-28T06:53:27.761713-05:
      DOI: 10.1002/jcp.25670
  • The Emerging Role of MORC Family Proteins in Cancer Development and Bone
    • Authors: Guoju Hong; Heng Qiu, Chao Wang, Gaurav Jadhav, Haibin Wang, Jennifer Tickner, Wei He, Jiake Xu
      Abstract: Microrchidia (MORC or MORC family CW-type zinc finger protein), a highly conserved nuclear protein superfamily, is an interesting new player in signaling-dependent chromatin remodeling and epigenetic regulation. MORC family proteins consist of MORC1, MORC2, MORC3 and MORC4 which display common structural determinants such as CW-type zinc finger and coiled-coil domains. They also exhibit unique structural motifs and tissue-specific expression profiles. Morc1 was first discovered as a key regulator for male meiosis and spermatogenesis. Accumulating biochemical and functional analyses unveil MORC proteins as key regulators for cancer development. More recently, using an ENU mutagenesis mouse model, MORC3 was found to play a role in regulating bone and calcium hemostasis. Here we discuss recent research progress on the emerging role of MORC proteins in cancer development and bone metabolism. Unravelling the cellular and molecular mechanisms by which MORC proteins carry out their functions in a tissue specific manner are important subjects for future investigation. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-28T06:46:21.015841-05:
      DOI: 10.1002/jcp.25665
  • Extracellular Matrix and Colorectal Cancer: How Surrounding
           Microenvironment Affects Cancer Cell Behavior?
    • Authors: Sara Crotti; Martina Piccoli, Flavio Rizzolio, Antonio Giordano, Donato Nitti, Marco Agostini
      Abstract: Colorectal cancer (CRC) whit more than a million of new cases per year is one of the most common registered cancers worldwide with few treatment options especially for advanced and metastatic patients.The tumor microenvironment is composed by extracellular matrix (ECM), cells and interstitial fluids. Among all these constituents, in the last years an increased interest around the ECM and its potential role in cancer tumorigenesis is arisen. During cancer progression the ECM structure and composition became disorganized, allowing cellular transformation and metastasis.Up to now, the focus has mainly been on the characterization of CRC microenvironment analyzing separately structural ECM components or cell secretome modifications. A more extensive view that interconnects these aspects should be addressed. In this review, biochemical (secretome) and biomechanical (structure and architecture) changes of tumor microenvironment will be discussed, giving suggestions on how these changes can affect cancer cell behavior. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-24T05:15:47.729256-05:
      DOI: 10.1002/jcp.25658
  • Valproic Acid and Lithium Meditate Anti-Inflammatory Effects by
           Differentially Modulating Dendritic Cell Differentiation and Function
    • Authors: Sy-Jye Leu; Yi-Yuan Yang, Hsing-Cheng Liu, Chieh-Yu Cheng, Yu-Chen Wu, Ming-Chyi Huang, Yuen-Lun Lee, Chi-Ching Chen, Winston W. Shen, Ko-Jiunn Liu
      Abstract: Valproic acid (VPA), with inhibition activity mainly towards histone deacetylase (HDAC) and Glycogen Synthase Kinase (GSK)-3, and lithium, with inhibition activity mainly toward GSK-3, are both prescribed in clinical as mood-stabilizers and anticonvulsants for the control of bipolar disorder. This study aims to compare the immuno-modulation activities of VPA and lithium, especially on the differentiation and functions of dendritic cells (DC). Our data show that treatment with VPA or lithium effectively alleviated the severity of collagen-induced arthritis triggered by LPS in mice. Both agents reduced the serum level of IL-6 and IL-10 after LPS challenge in mice. VPA and lithium both induce significant down-regulation of group I CD1 expression and secretion of IL-6 during differentiation of human monocyte-derived immature DC, while they differ in the induction of CD83 and CD86 expression, secretion of IL-8, IL-10, and TNF-α. Upon stimulation of immature DC with LPS, VPA and lithium both reduced the secretion of IL-6 and TNF-α. However, only lithium significantly increased the production of IL-10, while VPA increased the production of IL-8 but substantially reduce the secretion of IL-10 and IL-23. Treatment with VPA resulted in a reduced capacity of LPS-stimulated DC to promote the differentiation of T helper 17 cells that are critical in the promotion of inflammatory responses. Taken together, our results suggest that VPA and lithium may differentially modulate inflammation through regulating the capacity of DC to mediate distinct T cell responses, and they may provide a complementary immunomodulatory effects for the treatment of inflammation-related diseases. This article is protected by copyright. All rights reserved
      PubDate: 2016-09-17T14:30:50.294937-05:
      DOI: 10.1002/jcp.25604
  • Dermal-Epidermal Cross-Talk: Differential Interactions With Microvascular
           Endothelial Cells
    • Authors: Eleonora Bassino; Edoardo Vallariello, Franco Gasparri, Luca Munaron
      First page: 897
      Abstract: The biological importance of circulatory blood supply and angiogenesis for hair growth is now well recognized, but the their regulatory mechanisms require more mechanistic investigation. In vitro cocultures and tricultures can be successfully employed to greatly improve our knowledge on paracrine crosstalk between cell types that populate the dermal-epidermal interface and cutaneous vasculature. Here we report that human dermal fibroblasts (NHDF) promote viability and proliferation of microvascular endothelial cells (HMVEC), while HMVEC are not mitogenic for NHDF. In triculture setup, conditioned media obtained by cocultures (HMVEC/NHDF or HMVEC/follicle fibroblasts) differently modulate growth and proliferation of keratinocytes and alter the expression of metabolic and pro-inflammatory markers. In conclusion, tricultures were successfully employed to characterize in vitro dermal-epithelial and endothelial interactions and could integrate ex vivo and in vivo approaches by the use of high-throughput and standardized protocols in controlled conditions. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-20T18:36:10.658839-05:
      DOI: 10.1002/jcp.25657
  • Mitochondria-Targeted Antioxidants and Uncouplers of Oxidative
           Phosphorylation in Treatment of the Systemic Inflammatory Response
           Syndrome (SIRS)
    • Authors: Vlada V. Zakharova; Olga Yu. Pletjushkina, Roman A. Zinovkin, Ekaterina N. Popova, Boris V. Chernyak
      Pages: 904 - 912
      Abstract: Systemic inflammatory response syndrome (SIRS) development is accompanied by mitochondrial dysfunction and excessive ROS production. Mitochondrial dysfunctions also occur in many SIRS-related diseases and may be critical for their pathogenesis; therefore, a use of mitochondria-targeted drugs is a promising trend in SIRS research and therapy. Here, we review recent studies concerning the application of the mitochondria-targeted antioxidants and uncouplers of oxidative phosphorylation in animal models of SIRS and related diseases. We propose that a new class of uncouplers of oxidative phosphorylation, lipophilic cations could be a base for a new generation of drugs for SIRS treatment. J. Cell. Physiol. 232: 904–912, 2017. © 2016 Wiley Periodicals, Inc.Systemic inflammatory response syndrome (SIRS) development is accompanied by mitochondrial dysfunction and excessive ROS production; therefore, a use of mitochondria-targeted drugs is a promising trend in SIRS research and therapy. We propose that a new class of uncouplers of oxidative phosphorylation, lipophilic cations could be a base of a new generation of drugs for SIRS treatment.
      PubDate: 2016-10-17T07:51:14.576651-05:
      DOI: 10.1002/jcp.25626
  • Osteogenic Differentiation of Periosteal Cells during Fracture Healing
    • Authors: Tao Wang; Xinping Zhang, Daniel D. Bikle
      First page: 913
      Abstract: 5-10% of fractures fail to heal normally leading to additional surgery, morbidity, and altered quality of life. Fracture healing involves the coordinated action of stem cells primarily coming from the periosteum which differentiate into the chondrocytes and osteoblasts, forming first the soft (cartilage) callus followed by the hard (bone) callus. These stem cells are accompanied by a vascular invasion that appears critical for the differentiation process and which may enable the entry of osteoclasts necessary for the remodeling of the callus into mature bone. However, more research is needed to clarify the signaling events that activate the osteochondroprogenitor cells of periosteum and stimulate their differentiation into chondrocytes and osteoblasts. Ultimately a thorough understanding of the mechanisms for differential regulation of these osteochondroprogenitors will aid in the treatment of bone healing and the prevention of delayed union and nonunion of fractures. In this review, evidence supporting the concept that the periosteal cells are the major cell sources of skeletal progenitors for the fracture callus will be discussed. The osteogenic differentiation of periosteal cells manipulated by Wnt/β-catenin, TGF/BMP, Ihh/PTHrP, and IGF-1/PI3K–Akt signaling in fracture repair will be examined. The effect of physical (hypoxia and hyperoxia) and chemical factors (reactive oxygen species) as well as the potential coordinated regulatory mechanisms in the periosteal progenitor cells promoting osteogenic differentiation will also be discussed. Understanding the regulation of periosteal osteochondroprogenitors during fracture healing could provide insight into possible therapeutic targets and thereby help to enhance future fracture healing and bone tissue engineering approaches. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-12T06:40:55.646886-05:
      DOI: 10.1002/jcp.25641
  • The Natural cAMP Elevating Compound Forskolin in Cancer Therapy: Is it
    • Authors: Luigi Sapio; Monica Gallo, Michela Illiano, Emilio Chiosi, Daniele Naviglio, Annamaria Spina, Silvio Naviglio
      First page: 922
      Abstract: Cancer is a major public health problem and the second leading cause of mortality around the world. Although continuous advances in the science of oncology and cancer research are now leading to improved outcomes for many cancer patients, novel cancer treatment options are strongly demanded. Naturally occurring compounds from a variety of vegetables, fruits and medicinal plants have been shown to exhibit various anticancer properties in a number of in vitro and in vivo studies and represent an attractive research area for the development of new therapeutic strategies to fight cancer.Forskolin is a diterpene produced by the roots of the Indian plant Coleus forskohlii. The natural compound forskolin has been used for centuries in traditional medicine and its safety has also been documented in conventional modern medicine. Forskolin directly activates the adenylate cyclase enzyme, that generates cAMP from ATP, thus raising intracellular cAMP levels.Notably, cAMP signaling, through the PKA-dependent and/or independent pathways, is very relevant to cancer and its targeting has shown a number of antitumor effects, including the induction of mesenchymal-to-epithelial transition, inhibition of cell growth and migration and enhancement of sensitivity to conventional antitumor drugs in cancer cells.Here, we describe some features of cAMP signaling that are relevant to cancer biology and address the state of the art concerning the natural cAMP elevating compound forskolin and its perspectives as an effective anticancer agent. This article is protected by copyright. All rights reserved
      PubDate: 2016-10-14T03:21:46.56839-05:0
      DOI: 10.1002/jcp.25650
  • The Multifaceted Roles of Primary Cilia in the Regulation of Stem Cell
           Properties and Functions
    • Authors: Rui Lyu; Jun Zhou
      First page: 935
      Abstract: Stem cells are a unique class of cells that are capable of self-renewal and differentiation into multiple lineages. An increasing number of studies have suggested that both embryonic and adult stem cells possess primary cilia, antenna-like structures protruding from cell surfaces that are critical for sensing and transducing environmental cues. The primary cilium appears to regulate stem cells in multiple aspects, such as lineage specification and stemness maintenance. Understanding the role of primary cilia in the control of stem cell behavior could lead to the identification of new targets for regenerative therapies. Here, we discuss recent studies investigating the diverse roles of primary cilia in the regulation of stem cell properties and functions. We also propose potential new avenues for exploration in this promising field. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-11T07:05:29.536643-05:
      DOI: 10.1002/jcp.25683
  • Diverse Functions and Signal Transduction of the Exocyst Complex in Tumor
    • Authors: Toshiaki Tanaka; Kaoru Goto, Mitsuyoshi Iino
      First page: 939
      Abstract: The exocyst complex is a large conserved hetero-oligomeric complex that consists of Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84 subunits. It has been implicated in the targeting of vesicles for regulated exocytosis in various cell types, and is also important for targeted exocytosis of post-Golgi transport vesicles to the plasma membrane. The exocyst complex is essential for membrane growth, secretion, and function during exocytosis and endocytosis. Moreover, the individual components of the complex are thought to act on specific biological processes, such as cytokinesis, ciliogenesis, apoptosis, autophagy, and epithelial-mesenchymal transition (EMT). As a result, recent studies suggest that the exocyst complex may be involved in several diseases such as kidney disease, neuropathogenesis, diabetes, and cancer. In this review, we focus on the diverse functions and cellular signaling pathways of the exocyst complex in various tumors. J. Cell. Physiol. 9999: 1–19, 2016. © 2016 Wiley Periodicals, Inc.Exo70 interacts strongly with the Sec10–Sec15 subcomplex and Exo84 interacts with the Sec10–Sec15–Exo70 subcomplex. Sec3, Sec5, Sec6, and Sec8 form a tight quaternary subcomplex and Sec8 interacts with Sec10. Each component of the exocyst complex contributes to cancer progression and survival in diverse ways such as break of cell–cell contact, EMT, cell cycle progression, DNA repair, migration, autophagy, and anti-apoptosis.
      PubDate: 2016-12-22T06:30:54.740663-05:
      DOI: 10.1002/jcp.25619
  • Role of microRNAs on the Regulation of Mitochondrial Biogenesis and
           Insulin Signaling in Skeletal Muscle
    • Authors: Tanes I. Lima; Hygor N. Araujo, Eveline S. Menezes, Carlos H. Sponton, Michel B. Araújo, Lucas H.M. Bomfim, André L. Queiroz, Madla A. Passos, Thais Amaral e Sousa, Sandro M. Hirabara, Amanda R. Martins, Helena C.L.B. Sampaio, Alice Rodrigues, Rui Curi, Everardo M. Carneiro, Antônio C. Boschero, Leonardo R. Silveira
      Pages: 958 - 966
      Abstract: Mitochondria play a critical role in several cellular processes and cellular homeostasis. Mitochondrion dysfunction has been correlated with numerous metabolic diseases such as obesity and type 2 diabetes. MicroRNAs are non-coding RNAs that have emerged as key regulators of cell metabolism. The microRNAs act as central regulators of metabolic gene networks by leading to the degradation of their target messenger RNA or repression of protein translation. In addition, vesicular and non-vesicular circulating miRNAs exhibit a potential role as mediators of the cross-talk between the skeletal muscle and other tissues/organs. In this review, we will focus on the emerging knowledge of miRNAs controlling mitochondrial function and insulin signaling in skeletal muscle cells. J. Cell. Physiol. 232: 958–966, 2017. © 2016 Wiley Periodicals, Inc.Mitochondrion dysfunction in skeletal muscle has been correlated with innumerous metabolic diseases. MicroRNAs have emerged as key regulators of cell metabolism. In addition, vesicular and non-vesicular circulating miRNAs exhibit potential role as mediators of the cross-talking between skeletal muscle and other tissues controlling mitochondrial function and insulin signaling.
      PubDate: 2016-10-26T06:30:38.245457-05:
      DOI: 10.1002/jcp.25645
  • Tracing Males from Different Continents by Genotyping JC Polyomavirus in
           DNA from Semen Samples
    • Authors: John Charles Rotondo; Tommaso Candian, Rita Selvatici, Elisa Mazzoni, Gloria Bonaccorsi, Pantaleo Greco, Mauro Tognon, Fernanda Martini
      First page: 982
      Abstract: The human JC polyomavirus (JCPyV) is an ubiquitous viral agent infecting approximately 60% of humans. Recently, JCPyV sequences have been detected in semen samples. The aim of this investigation was to test whether semen JCPyV genotyping can be employed to trace the origin continent of males. Semen DNA samples (n = 170) from males of different Continents, were investigated by PCR for the polymorphic JCPyV viral capsid protein 1 (VP1) sequences, followed by DNA sequencing. JCPyV sequences were detected with an overall prevalence of 27.6% (47/170). DNA sequencing revealed that European males carried JCPyV types 1A (71.4%), 4 (11.4%), 2B (2.9%), 2D1 (2.9%) and 3A (2.9%), Asians JCPyV type 2D1 (66.7%) and Africans JCPyV types 3A (33.3%) and 1A (33.3%). In 10.6% of males, two different JCPyV genotypes were detected suggesting that the second JCPyV genotype was acquired in the destination country. This study indicates that the majority of semen samples, found to be JCPyV-positive, were infected with the JCPyV genotype found in the geographic area of male origin. Therefore, semen JCPyV genotyping could be employed to trace the origin continent of males. Our findings could be applied to forensic investigations, in case of, for instance, sexual crimes. Indeed, JCPyV genotyping should enable investigators to make additional detailed profiling of the offender. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-20T15:45:36.45259-05:0
      DOI: 10.1002/jcp.25686
  • Oligodendrocyte Progenitor Cells Directly Utilize Lactate for Promoting
           Cell Cycling and Differentiation
    • Authors: Yoshinori Ichihara; Toru Doi, Youngjae Ryu, Motoshi Nagao, Yasuhiro Sawada, Toru Ogata
      First page: 986
      Abstract: Oligodendrocyte progenitor cells (OPCs) undergo marked morphological changes to become mature oligodendrocytes, but the metabolic resources for this process have not been fully elucidated. Although lactate, a metabolic derivative of glycogen, has been reported to be consumed in oligodendrocytes as a metabolite, and to ameliorate hypomyelination induced by low glucose conditions, it is not clear about the direct contribution of lactate to cell cycling and differentiation of OPCs, and the source of lactate for remyelination. Therefore, we evaluated the effect of 1,4-dideoxy-1,4-imino-d-arabinitol (DAB), an inhibitor of the glycogen catabolic enzyme glycogen phosphorylase, in a mouse cuprizone model. Cuprizone induced demyelination in the corpus callosum and remyelination occurred after cuprizone treatment ceased. This remyelination was inhibited by the administration of DAB. To further examine whether lactate affects proliferation or differentiation of OPCs, we cultured mouse primary OPC-rich cells and analyzed the effect of lactate. Lactate rescued the slowed cell cycling induced by 0.4 mM glucose, as assessed by the BrdU-positive cell ratio. Lactate also promoted OPC differentiation detected by monitoring the mature oligodendrocyte marker myelin basic protein, in the presence of both 36.6 mM and 0.4 mM glucose. Furthermore, these lactate-mediated effects were suppressed by the reported monocarboxylate transporter inhibitor, α-cyano-4-hydroxy-cinnamate. These results suggest that lactate directly promotes the cell cycling rate and differentiation of OPCs, and that glycogen, one of the sources of lactate, contributes to remyelination in vivo. This article is protected by copyright. All rights reserved
      PubDate: 2016-11-15T03:31:01.611104-05:
      DOI: 10.1002/jcp.25690
  • Overexpression of Gremlin1 in Mesenchymal Stem Cells Improves Hindlimb
           Ischemia in Mice by Enhancing Cell Survival
    • Authors: Qiuling Xiang; Dongxi Hong, Yan Liao, Yong Cao, Muyun Liu, Jun Pang, Junjie Zhou, Guang Wang, Renhao Yang, Maosheng Wang, Andy Peng Xiang
      First page: 996
      Abstract: Mesenchymal stem cells (MSCs) are a promising cell resource for the treatment of ischemic diseases, partially through paracrine effects. One of the major obstacles of MSC treatment is the poor survival rate and low efficiency of transplanted stem cells due to ischemic or inflammatory environments. Gremlin1 (GREM1), a regulator of growth, differentiation and development, has been identified as a novel proangiogenic factor. However, the role and mechanism of GREM1 in MSCs remains unclear. Therefore, we assessed the putative beneficial effects of GREM1 on MSC-based therapy for hindlimb ischemia. The lentiviral vector, EF1a-GREM1, was constructed using the Multisite Gateway System and used to transduce MSCs. In vitro studies demonstrated increased survival of GREM1-MSCs exposed to H2O2, which is consistent with the activation of caspase-3. Conditional medium from GREM1-MSCs (GREM1-MSC-CM) increased the anti-apoptotic effects of human umbilical vein endothelial cells (HUVECs), and this effect was attenuated by treatment with the PI3K/Akt pathway inhibitor LY294002. MSCs modified with GREM1 could significantly increase blood perfusion of the ischemic hindlimb in vivo in a mouse model, which was correlated to improved MSC survival. This study demonstrates that overexpression of GREM1 in MSCs have greater therapeutic effects against ischemia compared with wild-type MSCs by enhancing the survival of MSCs and ECs, which may provide new tools for studies investigating the treatment of ischemic diseases. J. Cell. Physiol. 9999: 1–12, 2016. © 2016 Wiley Periodicals, Inc.This study demonstrates that overexpression of GREM1 in MSCs have greater therapeutic effects against ischemia compared with wild-type MSCs by enhancing the survival of MSCs and ECs, which may provide new tools for studies investigating the treatment of ischemic diseases.
      PubDate: 2016-12-20T05:11:06.645394-05:
      DOI: 10.1002/jcp.25578
  • Association Between Aerobic Exercise and Rosiglitazone Avoided the NAFLD
           and Liver Inflammation Exacerbated in PPAR-α Knockout Mice
    • Authors: Helena A.P. Batatinha; Edson A. Lima, Alexandre A.S. Teixeira, Camila O. Souza, Luana A. Biondo, Loreana S. Silveira, Fabio S. Lira, José C. Rosa Neto
      First page: 1008
      Abstract: Nonalcoholic fatty liver disease (NAFLD) is one of the main liver diseases today, and may progress to steatohepatitis, cirrhosis, and hepatocellular carcinoma. Some studies have shown the beneficial effects of aerobic exercise on reversing NAFLD. To verify whether chronic aerobic exercise improves the insulin resistance, liver inflammation, and steatohepatitis caused by a high fat diet (HF) and whether PPARα is involved in these actions. C57BL6 wild type (WT) and PPAR-α knockout (KO) mice were fed with a standard diet (SD) or HF during 12 weeks; the HF mice were trained on a treadmill during the last 8 weeks. Serum glucose and insulin tolerances, serum levels of aspartate aminotransferase, hepatic content of triacylglycerol, cytokines, gene expression, and protein expression were evaluated in all animals. Chronic exposure to HF diet increased triacylglycerol accumulation in the liver, leading to NAFLD, increased aminotransferase in the serum, increased peripheral insulin resistance, and higher adiposity index. Exercise reduced all these parameters in both animal genotypes. The liver lipid accumulation was not associated with inflammation; trained KO mice, however, presented a huge inflammatory response that was probably caused by a decrease in PPAR-γ expression. We conclude that exercise improved the damage caused by a HF independently of PPARα, apparently by a peripheral fatty acid oxidation in the skeletal muscle. We also found that the absence of PPARα together with exercise leads to a decrease in PPAR-γ and a huge inflammatory response. J. Cell. Physiol. 9999: 1–12, 2016. © 2016 Wiley Periodicals, Inc.Exercise training improved lipids toxicity in the liver; PPAR-alpha KO developed liver inflammation after training protocol Roziglitazone reduced liver inflammation independent of PPAR-alpha.
      PubDate: 2016-11-30T10:40:48.544758-05:
      DOI: 10.1002/jcp.25440
  • 3-Methylcholanthrene/Aryl-Hydrocarbon Receptor-Mediated Hypertension
           Through eNOS Inactivation
    • Authors: Chih-Cheng Chang; Yung-Ho Hsu, Hsiu-Chu Chou, Yuan-Chii G. Lee, Shu-Hui Juan
      Pages: 1020 - 1029
      Abstract: Endothelial nitric oxide synthase (eNOS) modulates vascular blood pressure and is predominantly expressed in endothelial cells and activated through the protein kinase B (Akt/PKB)-dependent pathway. We previously reported that 3-methylcholanthrene (3MC) activates the aryl hydrocarbon receptor (AhR) and reduces PI3K/Akt phosphorylation. This study investigated the mechanism underlying the downregulatory effects of 3-MC on nitric oxide (NO) production occurring through the AhR/RhoA/Akt-mediated mechanism. The mechanism underlying the effects of 3-MC on eNOS activity and blood pressure was examined in vitro and in vivo through genetic and pharmacological approaches. Results indicated that 3-MC modified heat shock protein 90 (HSP90), caveolin-1, dynein, and eNOS mRNA and protein expression through the AhR/RhoA-dependent mechanism in mouse cerebral vascular endothelial cells (MCVECs) and that 3-MC reduced eNOS phosphorylation through the AhR/RhoA-mediated inactivation of Akt1. The upregulation of dynein expression was associated with decreased eNOS dimer formation (eNOS dimer; an activated form of the enzyme). Coimmunoprecipitation assay results indicated that 3-MC significantly reduced the interaction between eNOS and its regulatory proteins, including Akt1 and HSP90, but increased the interaction between eNOS and caveolin-1. Immunofluorescence and Western blot analysis revealed that 3-MC reduced the amount of membrane-bound activated eNOS, and a modified Griess assay revealed that 3-MC concomitantly reduced NO production. However, simvastatin reduced 3-MC-mediated murine hypertension. Our study results indicate that AhR, RhoA, and eNOS have major roles in blood pressure regulation. Statin intervention might provide a potential therapeutic approach for reducing hypertension caused by 3-MC. J. Cell. Physiol. 232: 1020–1029, 2017. © 2016 Wiley Periodicals, Inc.Our study is the first to report that the AhR ligand 3-MC inactivated eNOS associated with downregulated HSP90 and upregulated caveolin-1 and dynein expression through AhR/RhoA-mediated PI3K/Akt inactivation, leading to hypertension in mice. The blockade of AhR and RhoA by using siAhR, DNRhoA, or statins reversed the inhibitory effects of 3-MC on eNOS by correcting the alterations of eNOS regulatory proteins.
      PubDate: 2016-08-10T00:29:24.094759-05:
      DOI: 10.1002/jcp.25497
  • MiR-145 Regulates Lipogenesis in Goat Mammary Cells Via Targeting INSIG1
           and Epigenetic Regulation of Lipid-Related Genes
    • Authors: Hui Wang; Huaiping Shi, Jun Luo, Yongqing Yi, Dawei Yao, Xueying Zhang, Gongzhen Ma, Juan J. Loor
      Pages: 1030 - 1040
      Abstract: MicroRNAs (miRNAs) are noncoding RNA molecules that regulate gene expression at the post-transcriptional level to cause translational repression or degradation of targets. The profiles of miRNAs across stages of lactation in small ruminant species such as dairy goats is unknown. A small RNA library was constructed using tissue samples from mammary gland of Saanen dairy goats harvested at mid-lactation followed by sequencing via Solexa technology. A total of 796 conserved miRNAs, 263 new miRNAs, and 821 pre-miRNAs were uncovered. After comparative analyses of our sequence data with published mammary gland transcriptome data across different stages of lactation, a total of 37 miRNAs (including miR-145) had significant differences in expression over the lactation cycle. Further studies revealed that miR-145 regulates metabolism of fatty acids in goat mammary gland epithelial cells (GMEC). Compared with nonlactating mammary tissue, lactating mammary gland had a marked increase in expression of miR-145. Overexpression of miR-145 increased transcription of genes associated with milk fat synthesis resulting in greater fat droplet formation, triacylglycerol accumulation, and proportion of unsaturated fatty acids. In contrast, silencing of miR-145 impaired fatty acid synthesis. Inhibition of miR-145 increased methylation levels of fatty acid synthase (FASN), stearoyl-CoA desaturase 1 (SCD1), peroxisome proliferator-activated receptor gamma (PPARG), and sterol regulatory element binding transcription factor 1 (SREBF1). Luciferase reporter assays confirmed that insulin induced gene 1 (INSIG1) is a direct target of miR-145. These findings underscore the need for further studies to evaluate the potential for targeting miR-145 for improving beneficial milk components in ruminant milk. J. Cell. Physiol. 232: 1030–1040, 2017. © 2016 Wiley Periodicals, Inc.Mammary lipogenesis is partly regulated by epigenetic mechanisms through miR-145 and INSIG1.
      PubDate: 2016-09-07T14:25:37.979785-05:
      DOI: 10.1002/jcp.25499
  • Chondrocyte Morphology in Stiff and Soft Agarose Gels and the Influence of
           Fetal Calf Serum
    • Authors: Asima Karim; Andrew C. Hall
      Pages: 1041 - 1052
      Abstract: Changes to chondrocyte volume/morphology may have deleterious effects on extracellular matrix (ECM) metabolism potentially leading to cartilage deterioration and osteoarthritis (OA). The factors controlling chondrocyte properties are poorly understood, however, pericellular matrix (PCM) weakening may be involved. We have studied the density, volume, morphology, and clustering of cultured bovine articular chondrocytes within stiff (2% w/v) and soft (0.2% w/v) three-dimensional agarose gels. Gels with encapsulated chondrocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM; fetal calf serum (FCS) 1–10%;380 mOsm) for up to 7 days. Chondrocytes were fluorescently labeled after 1, 3, and 7 days with 5-chloromethylfluorescein-diacetate (CMFDA) and propidium iodide (PI) or 1,5-bis{[2-(di-methylamino)ethyl]amino}-4,8-dihydroxyanthracene-9,10-dione (DRAQ5) to identify cytoplasmic space or DNA and imaged by confocal laser scanning microscopy (CLSM). Chondrocyte density, volume, morphology, and clustering were quantified using Volocity™ software. In stiff gels after 7 d with 10% FCS, chondrocyte density remained unaffected and morphology was relatively normal with occasional cytoplasmic processes. However, in soft gels by day 1, chondrocyte volume increased (P = 0.0058) and by day 7, density increased (P = 0.0080), along with the percentage of chondrocytes of abnormal morphology (P 
      PubDate: 2016-08-16T02:10:42.324137-05:
      DOI: 10.1002/jcp.25507
  • Functional Effects of Cigarette Smoke-Induced Changes in Airway Smooth
           Muscle Mitochondrial Morphology
    • Authors: Bharathi Aravamudan; Michael Thompson, Gary C. Sieck, Robert Vassallo, Christina M. Pabelick, Y. S. Prakash
      Pages: 1053 - 1068
      Abstract: Long-term exposure to cigarette smoke (CS) triggers airway hyperreactivity and remodeling, effects that involve airway smooth muscle (ASM). We previously showed that CS destabilizes the networked morphology of mitochondria in human ASM by regulating the expression of mitochondrial fission and fusion proteins via multiple signaling mechanisms. Emerging data link regulation of mitochondrial morphology to cellular structure and function. We hypothesized that CS-induced changes in ASM mitochondrial morphology detrimentally affect mitochondrial function, leading to CS effects on contractility and remodeling. Here, ASM cells were exposed to 1% cigarette smoke extract (CSE) for 48 h to alter mitochondrial fission/fusion, or by inhibiting the fission protein Drp1 or the fusion protein Mfn2. Mitochondrial function was assessed via changes in bioenergetics or altered rates of proliferation and apoptosis. Our results indicate that both exposure to CS and inhibition of mitochondrial fission/fusion proteins affect mitochondrial function (i.e., energy metabolism, proliferation, and apoptosis) in ASM cells. In vivo, the airways in mice chronically exposed to CS are thickened and fibrotic, and the expression of proteins involved in mitochondrial function is dramatically altered in the ASM of these mice. We conclude that CS-induced changes in mitochondrial morphology (fission/fusion balance) correlate with mitochondrial function, and thus may control ASM proliferation, which plays a central role in airway health. J. Cell. Physiol. 232: 1053–1068, 2017. © 2016 Wiley Periodicals, Inc.Long-term exposure to cigarette smoke (CS) triggers airway hyperreactivity and remodeling, effects that involve airway smooth muscle (ASM). Previous studies show that CS destabilizes the networked morphology of mitochondria in human ASM. We hypothesized that CS-induced changes in ASM mitochondrial morphology detrimentally affect mitochondrial function, leading to CS effects on contractility and remodeling.
      PubDate: 2016-09-21T10:15:28.976808-05:
      DOI: 10.1002/jcp.25508
  • Uric Acid Amplifies Aβ Amyloid Effects Involved in the Cognitive
           Dysfunction/Dementia: Evidences From an Experimental Model In Vitro
    • Authors: Giovambattista Desideri; Roberta Gentile, Andrea Antonosante, Elisabetta Benedetti, Davide Grassi, Loredana Cristiano, Antonello Manocchio, Sara Selli, Rodolfo Ippoliti, Claudio Ferri, Claudio Borghi, Antonio Giordano, Annamaria Cimini
      Pages: 1069 - 1078
      Abstract: There is still a considerable debate concerning whether uric acid is neuroprotective or neurotoxic agent. To clarify this topic, we tested the effects of uric acid on neuronal cells biology by using differentiated SHSY5Y neuroblastoma cells incubated with amyloid β to reproduce an in vitro model of Alzheimer's disease. The incubation of cells with uric acid at the dose of 40 µM or higher significantly reduced cell viability and potentiated the proapoptotic effect of amyloid β. Finally, uric acid enhanced the generation of 4-hydroxynonenal and the expression of PPARβ/δ promoted by amyloid β, indicating a prooxidant effects. In conclusion, uric acid could exert a detrimental influence on neuronal biology being this influence further potentiated by the concomitant exposure to neurotoxic stimuli. This effect is evident for uric acid concentrations close to those achievable in cerebrospinal fluid in presence of mild hyperuricemia thus suggesting a potential role of uric acid in pathophysiology of cognitive dysfunction. These effects are influenced by the concentrations of uric acid and by the presence of favoring conditions that commonly occur in neurodegenerative disorders and well as in the aging brain, including increased oxidative stress and exposure to amyloid β. J. Cell. Physiol. 232: 1069–1078, 2017. © 2016 Wiley Periodicals, Inc.The study suggests that uric acid could exert detrimental effects on brain structure and function by directly influencing the viability of neuronal cells and their ability to establish synaptic connections.
      PubDate: 2016-08-16T02:10:50.861306-05:
      DOI: 10.1002/jcp.25509
  • VIP Family Members Prevent Outer Blood Retinal Barrier Damage in a Model
           of Diabetic Macular Edema
    • Authors: Grazia Maugeri; Agata Grazia D'Amico, Caterina Gagliano, Salvatore Saccone, Concetta Federico, Sebastiano Cavallaro, Velia D'Agata
      Pages: 1079 - 1085
      Abstract: Diabetic macular edema (DME), characterized by an increase of thickness in the eye macular area, is due to breakdown of the blood-retinal barrier (BRB). Hypoxia plays a key role in the progression of this pathology by activating the hypoxia-inducible factors. In the last years, various studies have put their attention on the role of pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) in retinal dysfunction. However, until now, no study has investigated their protective role against the harmful combined effect of both hyperglycemia and hypoxia on outer BRB. Therefore, in the present study, we have analyzed the role of these peptides on permeability, restoration of tight junctions expression and inhibition of hyperglycemia/hypoxia-induced apoptosis, in an experimental in vitro model of outer BRB. Our results have demonstrated that the peptides' treatment have restored the integrity of outer BRB induced by cell exposure to hyperglycemia/hypoxia. Their effect is mediated through the activation of phosphoinositide 3 kinase (PI3K)/Akt and mammalian mitogen activated protein kinase/Erk kinase (MAPK/ERK) signaling pathways. In conclusion, our study further clarifies the mechanism through which PACAP and VIP perform the beneficial effect on retinal damage induced by hyperglycemic/hypoxic insult, responsible of DME progression. J. Cell. Physiol. 232: 1079–1085, 2017. © 2016 Wiley Periodicals, Inc.PACAP and VIP restore the integrity of outer BRB exposed to hyperglycemia/hypoxia. PACAP and VIP perform their beneficial effect on retinal damage through activation of PI3K/Akt and MAPK/ERK signaling cascades.
      PubDate: 2016-09-21T10:45:25.121149-05:
      DOI: 10.1002/jcp.25510
  • Effects of Conjugated Linoleic Acid Associated With Endurance Exercise on
           Muscle Fibres and Peroxisome Proliferator-Activated Receptor γ
           Coactivator 1 α Isoforms
    • Authors: Rosario Barone; Claudia Sangiorgi, Antonella Marino Gammazza, Daniela D'Amico, Monica Salerno, Francesco Cappello, Cristoforo Pomara, Giovanni Zummo, Felicia Farina, Valentina Di Felice, Filippo Macaluso
      Pages: 1086 - 1094
      Abstract: Conjugated linoleic acid (CLA) has been reported to improve muscle hypertrophy, steroidogenesis, physical activity, and endurance capacity in mice, although the molecular mechanisms of its actions are not completely understood. The aim of the present study was to identify whether CLA alters the expression of any of the peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) isoforms, and to evaluate the possible existence of fibre-type-specific hypertrophy in the gastrocnemius and plantaris muscles. Mice were randomly assigned to one of four groups: placebo sedentary, CLA sedentary, placebo trained, or CLA trained. The CLA groups were gavaged with 35 μl per day of Tonalin® FFA 80 food supplement containing CLA throughout the 6-week experimental period, whereas the placebo groups were gavaged with 35 μl sunflower oil each day. Each administered dose of CLA corresponded to approximately 0.7 g/kg or 0.5%, of the dietary daily intake. Trained groups ran 5 days per week on a Rota-Rod for 6 weeks at increasing speeds and durations. Mice were sacrificed by cervical dislocation and hind limb posterior muscle groups were dissected and used for histological and molecular analyses. Endurance training stimulated mitochondrial biogenesis by PGC1α isoforms (tot, α1, α2, and α3) but CLA supplementation did not stimulate PGC1α isoforms or mitochondrial biogenesis in trained or sedentary mice. In the plantaris muscle, CLA supplementation induced a fibre-type-specific hypertrophy of type IIx muscle fibres, which was associated with increased capillary density and was different from the fibre-type-specific hypertrophy induced by endurance exercise (of types I and IIb muscle fibres). J. Cell. Physiol. 232: 1086–1094, 2017. © 2016 Wiley Periodicals, Inc.CLA supplementation did not stimulate PGC1α isoforms or mitochondrial biogenesis in trained or sedentary mice. CLA supplementation induced a fibre-type-specific hypertrophy of type IIx muscle fibres.
      PubDate: 2016-09-21T10:15:23.022164-05:
      DOI: 10.1002/jcp.25511
  • Store-Operated Ca2+ Entry in Oocytes Modulate the Dynamics of
           IP3-Dependent Ca2+ Release From Oscillatory to Tonic
    • Authors: Raphaël Courjaret; Maya Dib, Khaled Machaca
      Pages: 1095 - 1103
      Abstract: Ca2+ signaling is ubiquitous and mediates various cellular functions encoded in its spatial, temporal, and amplitude features. Here, we investigate the role of store-operated Ca2+ entry (SOCE) in regulating the temporal dynamics of Ca2+ signals in Xenopus oocytes, which can be either oscillatory or tonic. Oscillatory Ca2+ release from intracellular stores is typically observed at physiological agonist concentration. When Ca2+ release leads to Ca2+ store depletion, this triggers the activation of SOCE that translates into a low-amplitude tonic Ca2+ signal. SOCE has also been implicated in fueling Ca2+ oscillations when activated at low levels. Here, we show that sustained SOCE activation in the presence of IP3 to gate IP3 receptors (IP3R) results in a pump-leak steady state across the endoplasmic reticulum (ER) membrane that inhibits Ca2+ oscillations and produces a tonic Ca2+ signal. Tonic signaling downstream of SOCE activation relies on focal Ca2+ entry through SOCE ER-plasma membrane (PM) junctions, Ca2+ uptake into the ER, followed by release through open IP3Rs at distant sites, a process we refer to as “Ca2+ teleporting.” Therefore, sustained SOCE activation in the presence of an IP3-dependent “leak” pathway at the ER membrane results in a switch from oscillatory to tonic Ca2+ signaling. J. Cell. Physiol. 232: 1095–1103, 2017. © 2016 Wiley Periodicals, Inc.Calcium influx through SOCE inhibits calcium oscillations and favors a tonic mode of signaling. This is achieved through focal calcium entry at SOCE clusters, followed by calcium uptake into the ER through SERCA and re-release through open IP3 Receptors. A process referred to as “calcium teleporting.”
      PubDate: 2016-09-07T14:25:44.356793-05:
      DOI: 10.1002/jcp.25513
  • Wnt3A Induces GSK-3β Phosphorylation and β-Catenin Accumulation
           Through RhoA/ROCK
    • Authors: Jae-Gyu Kim; Myoung-Ju Kim, Won-Ji Choi, Mi-Young Moon, Hee-Jun Kim, Jae-Yong Lee, Jaebong Kim, Sung-Chan Kim, Seung Goo Kang, Goo-Young Seo, Pyeung-Hyeun Kim, Jae-Bong Park
      First page: 1104
      Abstract: In canonical pathway, Wnt3A has been known to stabilize β-catenin through the dissociation between β-catenin and glycogen synthase kinase-3β (GSK-3β) that suppresses the phosphorylation and degradation of β-catenin. In non-canonical signaling pathway, Wnt was known to activate Rho GTPases and to induce cell migration. The cross-talk between canonical and non-canonical pathways by Wnt signaling; however, has not been fully elucidated. Here, we revealed that Wnt3A induces not only the phosphorylation of GSK-3β and accumulation of β-catenin but also RhoA activation in RAW264.7 and HEK293 cells. Notably, sh-RhoA and Tat-C3 abolished both the phosphorylation of GSK-3β and accumulation of β-catenin. Y27632, an inhibitor of Rho-associated coiled coil kinase (ROCK) and si-ROCK inhibited both GSK-3β phosphorylation and β-catenin accumulation. Furthermore, active domain of ROCK directly phosphorylated the purified recombinant GSK-3β in vitro. In addition, Wnt3A-induced cell proliferation and migration, which were inhibited by Tat-C3 and Y27632. Taken together, we propose the cross-talk between canonical and non-canonical signaling pathways of Wnt3A, which induces GSK-3β phosphorylation and β-catenin accumulation through RhoA and ROCK activation. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.Wnt3A activates RhoA and ROCK, which induces GSK-3beta phosphorylation and beta-catenin accumulation. RhoA and ROCK are involved in the regulation of cell proliferation and migration upon Wnt3A.
      PubDate: 2016-11-20T15:45:29.212161-05:
      DOI: 10.1002/jcp.25572
  • Early Growth Response 3 (Egr3) Contributes a Maintenance of C2C12 Myoblast
    • Authors: Mitsutoshi Kurosaka; Yuji Ogura, Toshiya Funabashi, Tatsuo Akema
      Pages: 1114 - 1122
      Abstract: Satellite cell proliferation is a crucially important process for adult myogenesis. However, its regulatory mechanisms remain unknown. Early growth response 3 (Egr3) is a zinc-finger transcription factor that regulates different cellular functions. Reportedly, Egr3 interacts with multiple signaling molecules that are also known to regulate satellite cell proliferation. Therefore, it is possible that Egr3 is involved in satellite cell proliferation. Results of this study have demonstrated that Egr3 transcript levels are upregulated in regenerating mouse skeletal muscle after cardiotoxin injury. Using C2C12 myoblast culture (a model of activated satellite cells), results show that inhibition of Egr3 by shRNA impairs the myoblast proliferation rate. Results also show reduction of NF-кB transcriptional activity in Egr3-inhibited cells. Inhibition of Egr3 is associated with an increase in annexin V+ cell fraction and apoptotic protein activity including caspase-3 and caspase-7, and Poly-ADP ribose polymerase. By contrast, the reduction of cellular proliferation by inhibition of Egr3 was partially recovered by treatment of pan-caspase inhibitor Z-VAD-FMK. Collectively, these results suggest that Egr3 is involved in myoblast proliferation by interaction with survival signaling. J. Cell. Physiol. 232: 1114–1122, 2017. © 2016 Wiley Periodicals, Inc.Egr3 maintains myoblast proliferation by stabilizing basal NF-κB activity. However, the inhibition of Egr3 results in activation of apoptotic signaling by suppressing NF-κB activity. The activation of apoptotic signaling is associated with proliferation defect in myoblast.
      PubDate: 2016-10-12T10:20:59.674051-05:
      DOI: 10.1002/jcp.25574
  • Endothelial STAT3 Activation Increases Vascular Leakage Through
           Downregulating Tight Junction Proteins: Implications for Diabetic
    • Authors: Jang-Hyuk Yun; Sung Wook Park, Kyung-Jin Kim, Jong-Sup Bae, Eun Hui Lee, Sun Ha Paek, Seung U. Kim, Sangkyu Ye, Jeong-Hun Kim, Chung-Hyun Cho
      Pages: 1123 - 1134
      Abstract: Vascular inflammation is characteristic feature of diabetic retinopathy. In diabetic retina, a variety of the pro-inflammatory cytokines are elevated and involved in endothelial dysfunction. STAT3 transcription factor has been implicated in mediating cytokine signaling during vascular inflammation. However, whether and how STAT3 is involved in the direct regulation of the endothelial permeability is currently undefined. Our studies revealed that IL-6-induced STAT3 activation increases retinal endothelial permeability and vascular leakage in retinas of mice through the reduced expression of the tight junction proteins ZO-1 and occludin. In a co-culture model with microglia and endothelial cells under a high glucose condition, the microglia-derived IL-6 induced STAT3 activation in the retinal endothelial cells, leading to increasing endothelial permeability. In addition, IL-6-induced STAT3 activation was independent of ROS generation in the retinal endothelial cells. Moreover, we demonstrated that STAT3 activation downregulates the ZO-1 and occludin levels and increases the endothelial permeability through the induction of VEGF production in retinal endothelial cells. These results suggest the potential importance of IL-6/STAT3 signaling in regulating endothelial permeability and provide a therapeutic target to prevent the pathology of diabetic retinopathy. J. Cell. Physiol. 232: 1123–1134, 2017. © 2016 Wiley Periodicals, Inc.Vascular inflammation is characteristic feature of diabetic retinopathy. We demonstrated that STAT3 in retinal endothelial cells plays a causative role in downregulating ZO-1 and occludin levels and increasing the endothelial permeability through VEGF production in retinal endothelial cells. These results suggest the potential importance of IL-6/STAT3 signaling in regulating endothelial permeability and provide a therapeutic target to prevent the pathology of diabetic retinopathy.
      PubDate: 2016-09-21T10:05:28.489171-05:
      DOI: 10.1002/jcp.25575
  • HMGB1 Inhibits Apoptosis Following MI and Induces Autophagy via mTORC1
    • Authors: Eleonora Foglio; Giovanni Puddighinu, Antonia Germani, Matteo A. Russo, Federica Limana
      Pages: 1135 - 1143
      Abstract: Exogenous High Mobility Group Box-1 protein (HMGB1) has been reported to protect the infarcted heart but the underlying mechanism is quite complex. In particular, its effect on ischemic cardiomyocytes has been poorly investigated. Aim of the present study was to verify whether and how autophagy and apoptosis were involved in HMGB1-induced heart repair following myocardial infarction (MI). HMGB1 (200 ng) or denatured HMGB1 were injected in the peri-infarcted region of mouse hearts following acute MI. Three days after treatment, an upregulation of autophagy was detected in infarcted HMGB1 treated hearts compared to controls. Specifically, HMGB1 induced autophagy by significantly upregulating the protein expression of LC3, Beclin-1, and Atg7 in the border zone. To gain further insights into the molecular mechanism of HMGB1-mediated autophagy, WB analysis were performed in cardiomyocytes isolated from 3 days infarcted hearts in the presence and in the absence of HMGB1 treatment. Results showed that upregulation of autophagy by HMGB1 treatment was potentially related to activation of AMP-activated protein kinase (AMPK) and inhibition of the mammalian target of rapamycin complex 1 (mTORC1). Accordingly, in these hearts, phospho-Akt signaling pathway was inhibited. The induction of autophagy was accompanied by reduced cardiomyocyte apoptotic rate and decreased expression levels of Bax/Bcl-2 and active caspase-3 in the border zone of 3 days infarcted mice following HMGB1 treatment. We report the first in vivo evidence that HMGB1 treatment in a murine model of acute MI might induce cardiomyocyte survival through attenuation of apoptosis and AMP-activated protein kinase-dependent autophagy. J. Cell. Physiol. 232: 1135–1143, 2017. © 2016 Wiley Periodicals, Inc.Exogenous High Mobility Group Box-1 protein (HMGB1) has been reported to protect the infarcted heart but the underlying mechanism is quite complex; in particular, its effect on ischemic cardiomyocytes has been poorly investigated. HMGB1 (200 ng) or denatured HMGB1 were injected in the peri-infarcted region of mouse hearts following acute MI. We report the first in vivo evidence that HMGB1 treatment in a murine model of acute MI might induce cardiomyocyte survival through attenuation of apoptosis and AMP-activated protein kinase-dependent autophagy.
      PubDate: 2016-09-21T10:55:22.031754-05:
      DOI: 10.1002/jcp.25576
  • Risk Differences Between Prediabetes And Diabetes According To Breast
           Cancer Molecular Subtypes
    • Authors: A. Crispo; L.S.A. Augustin, M. Grimaldi, F. Nocerino, A. Giudice, E. Cavalcanti, M. Di Bonito, G. Botti, M. De Laurentiis, M. Rinaldo, E. Esposito, G. Riccardi, A. Amore, M. Libra, G. Ciliberto, D.J.A. Jenkins, M. Montella
      Pages: 1144 - 1150
      Abstract: Hyperglycemia and hyperinsulinemia may play a role in breast carcinogenesis and prediabetes and diabetes have been associated with increased breast cancer (BC) risk. However, whether BC molecular subtypes may modify these associations is less clear. We therefore investigated these associations in all cases and by BC molecular subtypes among women living in Southern Italy. Cases were 557 patients with non-metastatic incident BC and controls were 592 outpatients enrolled during the same period as cases and in the same hospital for skin-related non-malignant conditions. Adjusted multivariate logistic regression models were built to assess the risks of developing BC in the presence of prediabetes or diabetes. The analyses were repeated by strata of BC molecular subtypes: Luminal A, Luminal B, HER2+, and Triple Negative (TN). Prediabetes and diabetes were significantly associated with higher BC incidence after controlling for known risk factors (OR = 1.94, 95% CI 1.32–2.87 and OR = 2.46, 95% CI 1.38–4.37, respectively). Similar results were seen in Luminal A and B while in the TN subtype only prediabetes was associated with BC (OR = 2.43, 95% CI 1.11–5.32). Among HER2+ patients, only diabetes was significantly associated with BC risk (OR = 3.04, 95% CI 1.24–7.47). Furthermore, when postmenopausal HER2+ was split into hormone receptor positive versus negative, the association with diabetes remained significant only in the former (OR = 5.13, 95% CI 1.53–17.22). These results suggest that prediabetes and diabetes are strongly associated with BC incidence and that these metabolic conditions may be more relevant in the presence of breast cancer molecular subtypes with positive hormone receptors. J. Cell. Physiol. 232: 1144–1150, 2017. © 2016 Wiley Periodicals, Inc.
      PubDate: 2016-09-30T08:12:46.992363-05:
      DOI: 10.1002/jcp.25579
  • Hyperexcitability of Mesencephalic Trigeminal Neurons and Reorganization
           of Ion Channel Expression in a Rett Syndrome Model
    • Authors: Max F. Oginsky; Ningren Cui, Weiwei Zhong, Christopher M. Johnson, Chun Jiang
      Pages: 1151 - 1164
      Abstract: People with Rett syndrome (RTT) have defects in motor function also seen in Mecp2-null mice. Motor function depends on not only central motor commands but also sensory feedback that is vulnerable to changes in excitability of propriosensory neurons. Here we report evidence for hyperexcitability of mesencephalic trigeminal (Me5) neurons in Mecp2-null mice and a novel cellular mechanism for lowering its impact. In in vitro brain slices, the Me5 neurons in both Mecp2−/Y male and symptomatic Mecp2+/− female mice were overly excitable showing increased firing activity in comparison to their wild-type (WT) male and asymptomatic counterparts. In Mecp2−/Y males, Me5 neurons showed a reduced firing threshold. Consistently, the steady-state activation of voltage-gated Na+ currents (INa) displayed a hyperpolarizing shift in the Mecp2-null neurons with no change in the INa density. This seems to be due to NaV1.1, SCN1B and SCN4B overexpression and NaV1.2 and SCN3B under-expression. In contrast to the hyperexcitability, the sag potential and postinhibitory rebound (PIR) were reduced in Mecp2-null mice. In voltage-clamp, the IH density was deficient by ∼33%, and the steady-state half-activation had a depolarizing shift of ∼10 mV in the Mecp2-null mice. Quantitative PCR analysis indicated that HCN2 was decreased, HCN1 was upregulated with no change in HCN4 in Mecp2−/Y mice compared to WT. Lastly, blocking IH reduced the firing rate much more in WT than in Mecp2-null neurons. These data suggest that the Mecp2 defect causes an increase in Me5 neuronal excitability likely attributable to alterations in INa, meanwhile IH is reduced likely altering neuronal excitability as well. J. Cell. Physiol. 232: 1151–1164, 2017. © 2016 Wiley Periodicals, Inc.Mesencephalic trigeminal V neurons are hyperexcitable in Mecp2-null mice. Our data suggest that the Mecp2 defect that causes an increase in Me5 neuronal excitability is likely attributable to alterations in INa, meanwhile IH is reduced likely altering neuronal excitability as well. Further, the changes in these currents seem to be due to alterations in channel subunit composition.
      PubDate: 2016-09-30T09:09:00.806058-05:
      DOI: 10.1002/jcp.25589
  • Inflammatory Events and Oxidant Production in the Diaphragm,
           Gastrocnemius, and Blood of Rats Exposed to Chronic Intermittent Hypoxia:
           Therapeutic Strategies
    • Authors: Marisol Domínguez-Álvarez; Joaquim Gea, Esther Barreiro
      Pages: 1165 - 1175
      Abstract: We hypothesized that inflammatory events and reactive oxygen species (ROS) production may be differentially expressed in respiratory and limb muscles, and blood of a chronic intermittent hypoxia (CIH) experimental model and that antioxidants and TNF-alpha blockade may influence those events. In blood, diaphragm, and gastrocnemius of rats non-invasively exposed to CIH (10% hypoxia, 2 h/day, 14 consecutive days) with/without concomitant treatment with either anti-TNF-alpha antibody (infliximab) or N-acetyl cysteine (NAC), inflammatory cytokines, superoxide anion production, muscle structural abnormalities, and fiber-type composition were assessed. Compared to non-exposed controls, in CIH-exposed rats, body weight gain was reduced, TNF-alpha, IL-1beta, IL-6, and interferon-gamma levels were increased in diaphragm, TNF-alpha, and IL-1 beta plasma levels were greater, systemic and muscle superoxide anion production was higher, diaphragm and gastrocnemius inflammatory cells and internal nuclei were higher, and muscle fiber-type and morphometry remained unmodified. CIH rats treated with infliximab further increased TNF-alpha, IL-1beta, IL-6, and interferon-gamma diaphragm levels, whereas NAC induced a reduction only in TNF-alpha and IL-1beta levels in diaphragm and plasma. Infliximab and NAC elicited a significant decline in superoxide anion production in diaphragm, gastrocnemius, and plasma, while inducing a further increase in inflammatory cells and internal nuclei in both muscles. Proinflammatory cytokines are differentially expressed in respiratory and limb muscles and plasma of CIH-exposed rats, while superoxide anion production increased in both muscle types and blood. Infliximab and NAC exerted different effects. These findings may help understand the biology underlying CIH in skeletal muscles and blood of patients with chronic respiratory diseases. J. Cell. Physiol. 232: 1165–1175, 2017. © 2016 Wiley Periodicals, Inc.Proinflammatory cytokines are differentially expressed in respiratory and limb muscles and plasma of CIH-exposed rats, while superoxide anion production increased in both muscle types and blood. Infliximab and NAC exerted different effects. These findings may help understand the biology underlying CIH in skeletal muscles and blood of patients with chronic respiratory diseases.
      PubDate: 2016-09-30T09:08:52.834967-05:
      DOI: 10.1002/jcp.25600
  • Interfering of the Reelin/ApoER2/PSD95 Signaling Axis Reactivates
           Dendritogenesis of Mature Hippocampal Neurons
    • Authors: Estibaliz Ampuero; Nur Jury, Steffen Härtel, María-Paz Marzolo, Brigitte van Zundert
      Pages: 1187 - 1199
      Abstract: Reelin, an extracellular glycoprotein secreted in embryonic and adult brain, participates in neuronal migration and neuronal plasticity. Extensive evidence shows that reelin via activation of the ApoER2 and VLDLR receptors promotes dendrite and spine formation during early development. Further evidence suggests that reelin signaling is needed to maintain a stable architecture in mature neurons, but, direct evidence is lacking. During activity-dependent maturation of the neuronal circuitry, the synaptic protein PSD95 is inserted into the postsynaptic membrane to induce structural refinement and stability of spines and dendrites. Given that ApoER2 interacts with PSD95, we tested if reelin signaling interference in adult neurons reactivates the dendritic architecture. Unlike findings in developing cultures, the presently obtained in vitro and in vivo data show, for the first time, that reelin signaling interference robustly increase dendritogenesis and reduce spine density in mature hippocampal neurons. In particular, the expression of a mutant ApoER2 form (ApoER2-tailless), which is unable to interact with PSD95 and hence cannot transduce reelin signaling, resulted in robust dendritogenesis in mature hippocampal neurons in vitro. These results indicate that reelin/ApoER2/PSD95 signaling is important for neuronal structure maintenance in mature neurons. Mechanistically, obtained immunofluorescent data indicate that reelin signaling impairment reduced synaptic PSD95 levels, consequently leading to synaptic re-insertion of NR2B-NMDARs. Our findings underscore the importance of reelin in maintaining adult network stability and reveal a new mode for reactivating dendritogenesis in neurological disorders where dendritic arbor complexity is limited, such as in depression, Alzheimer's disease, and stroke. J. Cell. Physiol. 232: 1187–1199, 2017. © 2016 Wiley Periodicals, Inc.In mature neurons the reelin interference reactivates the dendritogenesis in the adults reelin signaling maintains adult network stability.
      PubDate: 2016-10-25T09:50:45.334588-05:
      DOI: 10.1002/jcp.25605
  • Pulsed Electromagnetic Field Exposure Reduces Hypoxia and Inflammation
           Damage in Neuron-Like and Microglial Cells
    • Authors: Fabrizio Vincenzi; Annalisa Ravani, Silvia Pasquini, Stefania Merighi, Stefania Gessi, Stefania Setti, Ruggero Cadossi, Pier Andrea Borea, Katia Varani
      Pages: 1200 - 1208
      Abstract: In the present study, the effect of low-frequency, low-energy pulsed electromagnetic fields (PEMFs) has been investigated by using different cell lines derived from neuron-like cells and microglial cells. In particular, the primary aim was to evaluate the effect of PEMF exposure in inflammation- and hypoxia-induced injury in two different neuronal cell models, the human neuroblastoma-derived SH-SY5Y cells and rat pheochromocytoma PC12 cells and in N9 microglial cells. In neuron-like cells, live/dead and apoptosis assays were performed in hypoxia conditions from 2 to 48 h. Interestingly, PEMF exposure counteracted hypoxia damage significantly reducing cell death and apoptosis. In the same cell lines, PEMFs inhibited the activation of the hypoxia-inducible factor 1α (HIF-1α), the master transcriptional regulator of cellular response to hypoxia. The effect of PEMF exposure on reactive oxygen species (ROS) production in both neuron-like and microglial cells was investigated considering their key role in ischemic injury. PEMFs significantly decreased hypoxia-induced ROS generation in PC12, SH-SY5Y, and N9 cells after 24 or 48 h of incubation. Moreover, PEMFs were able to reduce some of the most well-known pro-inflammatory cytokines such as tumor necrosis factor–α (TNF-α), interleukin (IL)-1β, IL-6, and IL-8 release in N9 microglial cells stimulated with different concentrations of LPS for 24 or 48 h of incubation time. These results show a protective effect of PEMFs on hypoxia damage in neuron-like cells and an anti-inflammatory effect in microglial cells suggesting that PEMFs could represent a potential therapeutic approach in cerebral ischemic conditions. J. Cell. Physiol. 232: 1200–1208, 2017. © 2016 Wiley Periodicals, Inc.In the present study, the effect of low-frequency, low-energy pulsed electromagnetic fields (PEMFs) has been investigated by using different cell lines derived from neuron-like cells and microglial cells. PEMF exposure counteracted hypoxia damage significantly reducing cell death and apoptosis, reduced HIF-1α, ROS, and proinflammatory cytokines. PEMFs could represent a potential therapeutic approach in cerebral ischemic conditions.
      PubDate: 2016-10-07T05:56:35.233678-05:
      DOI: 10.1002/jcp.25606
  • PACAP and VIP Inhibit HIF-1α-Mediated VEGF Expression in a Model of
           Diabetic Macular Edema
    • Authors: Grazia Maugeri; Agata Grazia D'Amico, Salvatore Saccone, Concetta Federico, Sebastiano Cavallaro, Velia D'Agata
      Pages: 1209 - 1215
      Abstract: Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) exert a protective role against retinal injuries, including diabetic macular edema (DME). The macular damage is induced by hyperglycemia, which damages vessels supplying blood to the retina and induces hypoxia. The microenvironmental changes stimulate the expression of hypoxia-inducible factors (HIFs), which promote the choroidal endothelial cell transmigration across the retinal pigmented epithelium (RPE) into neurosensory retina, where they proliferate into new vessels under stimulation of the vascular endothelial growth factor (VEGF). In the present study, we have investigated whether PACAP and VIP prevent retinal damage by modulating the expression of HIFs, VEGF, and its receptors. In accord to our hypothesis, we have shown that both peptides are able to significantly reduce HIF-1α and increase HIF-3α expression in ARPE-19 cells exposed to hyperglycemic/hypoxic insult. This effect is also related to a reduction of VEGF and its receptors expression. Moreover, both peptides also reduce the activation of p38 mitogen-activated protein kinase (MAPK), a pro-apoptotic signaling pathway, which is activated by VEGFR-1 and 2 receptors. In conclusion, our study has further elucidated the protective role performed by PACAP and VIP, against the harmful combined effect of hyperglycemia/hypoxia characterizing the DME microenvironment. J. Cell. Physiol. 232: 1209–1215, 2017. © 2016 Wiley Periodicals, Inc.PACAP and VIP prevent retinal damage by modulating the expression of HIFs, VEGF, and its receptors; PACAP and VIP act against the harmful combined effect of hyperglycemia/hypoxia.
      PubDate: 2016-09-30T09:16:20.837141-05:
      DOI: 10.1002/jcp.25616
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