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https://www.readbyqxmd.com/read/28231263/uniformity-under-in-vitro-conditions-changes-in-the-phenotype-of-cancer-cell-lines-derived-from-different-medulloblastoma-subgroups
#1
Petr Chlapek, Karel Zitterbart, Leos Kren, Lenka Filipova, Jaroslav Sterba, Renata Veselska
Medulloblastoma comprises four main subgroups (WNT, SHH, Group 3 and Group 4) originally defined by transcriptional profiling. In primary medulloblastoma tissues, these groups are thought to be distinguishable using the immunohistochemical detection of β-catenin, filamin A, GAB1 and YAP1 protein markers. To investigate the utility of these markers for in vitro studies using medulloblastoma cell lines, immunoblotting and indirect immunofluorescence were employed for the detection of β-catenin, filamin A, GAB1 and YAP1 in both DAOY and D283 Med reference cell lines and the panel of six medulloblastoma cell lines derived in our laboratory from the primary tumor tissues of known molecular subgroups...
2017: PloS One
https://www.readbyqxmd.com/read/28225808/epileptogenesis-in-organotypic-hippocampal-cultures-has-limited-dependence-on-culture-medium-composition
#2
Jing Liu, Yero Saponjian, Mark M Mahoney, Kevin J Staley, Yevgeny Berdichevsky
Rodent organotypic hippocampal cultures spontaneously develop epileptiform activity after approximately 2 weeks in vitro and are increasingly used as a model of chronic post-traumatic epilepsy. However, organotypic cultures are maintained in an artificial environment (culture medium), which contains electrolytes, glucose, amino acids and other components that are not present at the same concentrations in cerebrospinal fluid (CSF). Therefore, it is possible that epileptogenesis in organotypic cultures is driven by these components...
2017: PloS One
https://www.readbyqxmd.com/read/28219951/generation-of-thalamic-neurons-from-mouse-embryonic-stem-cells
#3
Atsushi Shiraishi, Keiko Muguruma, Yoshiki Sasai
Thalamus is a diencephalic structure that plays crucial roles in relaying and modulating sensory and motor information to the neocortex. The thalamus develops in the dorsal part of the neural tube at the level of the caudal forebrain. However, the molecular mechanisms that are essential for thalamic differentiation are still unknown. Here we have succeeded in the generation of thalamic neurons from mouse ES cells (mESCs) by modifying the default method that induces the most-anterior neural type in self-organizing culture...
February 20, 2017: Development
https://www.readbyqxmd.com/read/28210712/identification-of-an-epidermal-keratinocyte-ampa-glutamate-receptor-involved-in-dermatopathies-associated-with-sensory-abnormalities
#4
David Cabañero, Takeshi Irie, Marta Celorrio, Christopher Trousdale, David M Owens, David Virley, Phillip J Albrecht, Michael J Caterina, Frank L Rice, Jose A Morón
INTRODUCTION: Epidermal keratinocytes are increasingly recognized as active participants in the sensory transduction of itch and pain, processes known to involve primary afferent glutamatergic neurons. However the role of keratinocyte glutamate signaling in sensory functioning is not fully understood. Here, we present the observation of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid type glutamate receptors (AMPAR) in epidermal keratinocytes. METHODS: Immunohistochemical and in situ hybridization analyses were conducted to assess the expression of AMPAR subunits in epidermal keratinocytes in mouse and human skin samples, and in organotypic cultures of human keratinocytes...
September 2016: Pain Reports (Baltimore, Md.)
https://www.readbyqxmd.com/read/28209528/decellularized-human-colorectal-cancer-matrices-polarize-macrophages-towards-an-anti-inflammatory-phenotype-promoting-cancer-cell-invasion-via-ccl18
#5
M L Pinto, E Rios, A C Silva, S C Neves, H R Caires, A T Pinto, C Durães, F A Carvalho, A P Cardoso, N C Santos, C C Barrias, D S Nascimento, P Pinto-do-Ó, M A Barbosa, F Carneiro, M J Oliveira
Macrophages are frequently identified in solid tumors, playing important roles in cancer progression. Their remarkable plasticity makes them very sensitive to environmental factors, including the extracellular matrix (ECM). In the present work, we investigated the impact of human colorectal tumor matrices on macrophage polarization and on macrophage-mediated cancer cell invasion. Accordingly, we developed an innovative 3D-organotypic model, based on the decellularization of normal and tumor tissues derived from colorectal cancer patients' surgical resections...
February 5, 2017: Biomaterials
https://www.readbyqxmd.com/read/28205613/pak4-interacts-with-p85-alpha-implications-for-pancreatic-cancer-cell-migration
#6
Helen King, Kiruthikah Thillai, Andrew Whale, Prabhu Arumugam, Hesham Eldaly, Hemant M Kocher, Claire M Wells
It has been reported that p21-activated kinase 4 (PAK4) is amplified in pancreatic cancer tissue. PAK4 is a member of the PAK family of serine/threonine kinases, which act as effectors for several small GTPases, and has been specifically identified to function downstream of HGF-mediated c-Met activation in a PI3K dependent manner. However, the functionality of PAK4 in pancreatic cancer and the contribution made by HGF signalling to pancreatic cancer cell motility remain to be elucidated. We now find that elevated PAK4 expression is coincident with increased expression levels of c-Met and the p85α subunit of PI3K...
February 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28199128/sox2-drives-bronchial-dysplasia-in-a-novel-organotypic-model-of-early-human-squamous-lung-cancer
#7
Lúcia L Correia, Jo-Anne Johnson, Peter McErlean, Julien Bauer, Hassan Farah, Doris M Rassl, Robert C Rintoul, Tariq Sethi, Paul Lavender, Emma L Rawlins, Trevor D Littlewood, Gerard I Evan, Frank M McCaughan
Rationale Improving the early detection and chemoprevention of lung cancer are key to improving outcomes. The pathobiology of early squamous lung cancer is poorly understood. We have shown that amplification of SOX2 is an early and consistent event in the pathogenesis of this disease but its functional oncogenic potential remains uncertain. We tested the impact of deregulated SOX2 expression in a novel organotypic system that recreates the molecular and microenvironmental context in which squamous carcinogenesis occurs...
February 15, 2017: American Journal of Respiratory and Critical Care Medicine
https://www.readbyqxmd.com/read/28197070/role-of-app-interactions-with-heterotrimeric-g-proteins-physiological-functions-and-pathological-consequences
#8
REVIEW
Philip F Copenhaver, Donat Kögel
Following the discovery that the amyloid precursor protein (APP) is the source of β-amyloid peptides (Aβ) that accumulate in Alzheimer's disease (AD), structural analyses suggested that the holoprotein resembles a transmembrane receptor. Initial studies using reconstituted membranes demonstrated that APP can directly interact with the heterotrimeric G protein Gαo (but not other G proteins) via an evolutionarily G protein-binding motif in its cytoplasmic domain. Subsequent investigations in cell culture showed that antibodies against the extracellular domain of APP could stimulate Gαo activity, presumably mimicking endogenous APP ligands...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28194096/morphological-plasticity-of-emerging-purkinje-cells-in-response-to-exogenous-vegf
#9
Leonard Herrfurth, Verena Theis, Veronika Matschke, Caroline May, Katrin Marcus, Carsten Theiss
Vascular endothelial growth factor (VEGF) is well known as the growth factor with wide-ranging functions even in the central nervous system (CNS). Presently, most attention is given to the investigation of its role in neuronal protection, growth and maturation processes, whereby most effects are mediated through VEGF receptor 2 (VEGFR-2). The purpose of our current study is to provide new insights into the impact of VEGF on immature and mature Purkinje cells (PCs) in accordance with maturity and related receptor expression...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28193830/il-6-regulates-m2-polarization-and-local-proliferation-of-adipose-tissue-macrophages-in-obesity
#10
Julia Braune, Ulrike Weyer, Constance Hobusch, Jan Mauer, Jens C Brüning, Ingo Bechmann, Martin Gericke
Obesity is associated with chronic low-grade inflammation of adipose tissue (AT) and an increase of AT macrophages (ATMs) that is linked to the onset of type 2 diabetes. We have recently shown that focal sites of inflammation around dying adipocytes, so-called crown-like structures, exhibit a unique microenvironment for macrophage proliferation. Interestingly, locally proliferating macrophages were not classically activated (M1), but they exhibited a rather alternatively activated (M2) immune phenotype. In this study, we established organotypic cell cultures of AT explants to study the impact of cytokine treatment on local ATM proliferation, without the bias of early monocyte recruitment...
February 13, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28191776/clinical-grade-isolated-human-kidney-perivascular-stromal-cells-as-an-organotypic-cell-source-for-kidney-regenerative-medicine
#11
Daniëlle G Leuning, Marlies E J Reinders, Joan Li, Anna J Peired, Ellen Lievers, Hetty C de Boer, Willem E Fibbe, Paola Romagnani, Cees van Kooten, Melissa H Little, Marten A Engelse, Ton J Rabelink
Mesenchymal stromal cells (MSCs) are immunomodulatory and tissue homeostatic cells that have shown beneficial effects in kidney diseases and transplantation. Perivascular stromal cells (PSCs) identified within several different organs share characteristics of bone marrow-derived MSCs (BM-MSCs). These PSCs may also possess tissue-specific properties and play a role in local tissue homeostasis. We hypothesized that human kidney-derived PSCs (hkPSCs) would elicit improved kidney repair in comparison with BM-MSCs...
February 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28184187/ankyrin-g-membrane-partners-drive-the-establishment-and-maintenance-of-the-axon-initial-segment
#12
Christophe Leterrier, Nadine Clerc, Fanny Rueda-Boroni, Audrey Montersino, Bénédicte Dargent, Francis Castets
The axon initial segment (AIS) is a highly specialized neuronal compartment that plays a key role in neuronal development and excitability. It concentrates multiple membrane proteins such as ion channels and cell adhesion molecules (CAMs) that are recruited to the AIS by the scaffold protein ankyrin G (ankG). The crucial function of ankG in the anchoring of AIS membrane components is well established, but a reciprocal role of membrane partners in ankG targeting and stabilization remained elusive. In rat cultured hippocampal neurons and cortical organotypic slices, we found that shRNA-mediated knockdown of ankG membrane partners (voltage-gated sodium channels (Nav) or neurofascin-186) led to a decrease of ankG concentration and perturbed the AIS formation and maintenance...
2017: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/28179525/differentiation-dependent-lmp1-expression-is-required-for-efficient-lytic-ebv-reactivation-in-epithelial-cells
#13
Dhananjay M Nawandar, Makoto Ohashi, Reza Djavadian, Elizabeth Barlow, Kathleen Makielski, Ahmed Ali, Denis Lee, Paul F Lambert, Eric Johannsen, Shannon C Kenney
Epstein-Barr virus (EBV)-associated diseases of epithelial cells, including tumors such as nasopharyngeal carcinoma (NPC) that have latent infection, and oral hairy leukoplakia (OHL) lesions that have lytic infection, frequently express the viral latent membrane protein 1 (LMP1). In lytically-infected cells, LMP1 expression is activated by the BRLF1 (R) immediate-early (IE) protein. However, the mechanisms by which LMP1 expression is normally regulated in epithelial cells remain poorly understood, and its potential roles in regulating lytic reactivation in epithelial cells are as yet unexplored...
February 8, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28174247/self-organisation-after-embryonic-kidney-dissociation-is-driven-via-selective-adhesion-of-ureteric-epithelial-cells
#14
J G Lefevre, H S Chiu, A N Combes, J M Vanslambrouck, A Ju, N A Hamilton, M H Little
Human pluripotent stem cells, after directed differentiation in vitro, can spontaneously generate complex tissues via self-organisation of the component cells. Self-organisation can also reform embryonic organ structure after tissue disruption. It has previously been demonstrated that dissociated embryonic kidneys can recreate component epithelial and mesenchymal relationships sufficient to allow continued kidney morphogenesis. Here we investigate the timing and underlying mechanisms driving self-organisation after dissociation of the embryonic kidney using time-lapse imaging, high-resolution confocal analyses and mathematical modelling...
February 7, 2017: Development
https://www.readbyqxmd.com/read/28173980/expression-of-polymeric-immunoglobulin-receptor-and-stromal-activity-in-pancreatic-ductal-adenocarcinoma
#15
Prabhu Arumugam, Satyajit Bhattacharya, Joanne Chin-Aleong, Melania Capaso, Hemant M Kocher
BACKGROUND/OBJECTIVES: Polymeric immunoglobulin receptor (pIgR) traffics Immunoglobulins (IgA and IgM) through epithelial cells in normal mucosae but neither are expressed in the normal pancreas. Recent work from our laboratory suggested pIgR may be upregulated in pancreatic ductal adenocarcinoma (PDAC). Our aim was to assess the role of pIgR in human PDAC. METHODS: pIgR expression was manipulated (siRNA and shRNA) in cell lines to evaluate its subsequent effect on cell behaviour in 2D assays as well as 3D organotypics models...
February 1, 2017: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
https://www.readbyqxmd.com/read/28167853/curcumin-alters-neural-plasticity-and-viability-of-intact-hippocampal-circuits-and-attenuates-behavioral-despair-and-cox-2-expression-in-chronically-stressed-rats
#16
Ga-Young Choi, Hyun-Bum Kim, Eun-Sang Hwang, Seok Lee, Min-Ji Kim, Ji-Young Choi, Sung-Ok Lee, Sang-Seong Kim, Ji-Ho Park
Curcumin is a major diarylheptanoid component of Curcuma longa with traditional usage for anxiety and depression. It has been known for the anti-inflammatory, antistress, and neurotropic effects. Here we examined curcumin effect in neural plasticity and cell viability. 60-channel multielectrode array was applied on organotypic hippocampal slice cultures (OHSCs) to monitor the effect of 10 μM curcumin in long-term depression (LTD) through low-frequency stimulation (LFS) to the Schaffer collaterals and commissural pathways...
2017: Mediators of Inflammation
https://www.readbyqxmd.com/read/28167635/defined-engineered-human-myocardium-with-advanced-maturation-for-applications-in-heart-failure-modelling-and-repair
#17
Malte Tiburcy, James E Hudson, Paul Balfanz, Susanne F Schlick, Tim Meyer, Mei-Ling Chang Liao, Elif Levent, Farah Raad, Sebastian Zeidler, Edgar Wingender, Johannes Riegler, Mouer Wang, Joseph D Gold, Itzhak Kehat, Erich Wettwer, Ursula Ravens, Pieterjan Dierickx, Linda van Laake, Marie-José Goumans, Sara Khadjeh, Karl Toischer, Gerd Hasenfuss, Larry A Couture, Andreas Unger, Wolfgang A Linke, Toshiyuki Araki, Benjamin Neel, Gordon Keller, Lior Gepstein, Joseph C Wu, Wolfram-Hubertus Zimmermann
BACKGROUND: -Advancing structural and functional maturation of stem cell-derived cardiomyocytes remains a key challenge for applications in disease modelling, drug screening, and heart repair. Here, we sought to advance cardiomyocyte maturation in engineered human myocardium (EHM) towards an adult phenotype under defined conditions. METHODS: -We systematically investigated cell composition, matrix and media conditions to generate EHM from embryonic and induced pluripotent stem cell-derived cardiomyocytes and fibroblasts with organotypic functionality under serum-free conditions...
February 6, 2017: Circulation
https://www.readbyqxmd.com/read/28165475/a-synthetic-aav-vector-enables-safe-and-efficient-gene-transfer-to-the-mammalian-inner-ear
#18
Lukas D Landegger, Bifeng Pan, Charles Askew, Sarah J Wassmer, Sarah D Gluck, Alice Galvin, Ruth Taylor, Andrew Forge, Konstantina M Stankovic, Jeffrey R Holt, Luk H Vandenberghe
Efforts to develop gene therapies for hearing loss have been hampered by the lack of safe, efficient, and clinically relevant delivery modalities. Here we demonstrate the safety and efficiency of Anc80L65, a rationally designed synthetic vector, for transgene delivery to the mouse cochlea. Ex vivo transduction of mouse organotypic explants identified Anc80L65 from a set of other adeno-associated virus (AAV) vectors as a potent vector for the cochlear cell targets. Round window membrane injection resulted in highly efficient transduction of inner and outer hair cells in mice, a substantial improvement over conventional AAV vectors...
February 6, 2017: Nature Biotechnology
https://www.readbyqxmd.com/read/28148885/stimulating-neurons-with-heterologously-expressed-light-gated-ion-channels
#19
J Simon Wiegert, Christine E Gee, Thomas G Oertner
Heterologous expression of ion channels that can be directly gated by light has made it possible to stimulate almost any excitable cell with light. Optogenetic stimulation has been particularly powerful in the neurosciences, as it allows the activation of specific, genetically defined neurons with precise timing. Organotypic hippocampal slice cultures are a favored preparation for optogenetic experiments. They can be cultured for many weeks and, after transfection with optogenetic actuators and sensors, allow the study of individual synapses or small networks...
February 1, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28148856/single-cell-electroporation-of-neurons
#20
J Simon Wiegert, Christine E Gee, Thomas G Oertner
Single-cell electroporation allows the transfection of a small number of neurons in an organotypic culture with a single plasmid or a defined mixture of plasmids. Desired protein expression levels can vary depending on the experimental goals (e.g., high expression levels are needed for optogenetic experiments); however, when too much protein is expressed, cellular toxicity and cell death may arise. To a large degree, protein expression can be controlled by adjusting the concentration of plasmid DNA in the electroporation pipette...
February 1, 2017: Cold Spring Harbor Protocols
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