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X Y Ji, J Shi, X X Dai, Y J Sheng, Y P Xue, J C Liu, H H Cai, X L Dai, Y M Chen, Y S Zhang, Q Huang, J Dong
Objective: A variety of interstitial cells in tumor microenvironment (TME) based on glioma stem cells(GSC) have the function to promote malignant progression of tumors, but whether these interstitial cells have already undergone malignant transformation and their related molecular characteristics are still poorly understood. Methods: Human SU3-RFP glioma stem cells (GSC) stably transfected with red fluorescent protein (RFP) and interstitial cells from enhanced green fluorescent protein (EGFP) transgenic nude mice were co-cultured in vitro...
November 6, 2018: Zhonghua Yi Xue za Zhi [Chinese medical journal]
Sandra Schneider, Nathan Hotaling, Maria Campos, Sarita Rani Patnaik, Kapil Bharti, Helen Louise May-Simera
The retinal pigment epithelium (RPE) is an epithelial monolayer in the back of the vertebrate eye. RPE dysfunction is associated with retinal degeneration and blindness. In order to fully understand how dysregulation affects visual function, RPE-specific gene knockouts are indispensable. Since the currently available RPE-specific Cre recombinases show lack of specificity or poor recombination, we sought to generate an alternative. We generated a tamoxifen-inducible RPE-specific Cre transgenic mouse line under transcriptional control of an RPE-specific Tyrosinase enhancer...
2018: PloS One
Koichi Matsuo, Ji Shuting, Ayako Miya, Masaki Yoda, Yuzuru Hamada, Tomoya Tanaka, Ryoko Takao-Kawabata, Katsuhiro Kawaai, Yukiko Kuroda, Shinsuke Shibata
The periosteum and mineralized bone are innervated by nerves that sense pain. These include both myelinated and unmyelinated neurons with either free nerve endings or bearing nociceptors. Parasympathetic and sympathetic autonomic nerves also innervate bone. However, little is known about the route sensory nerves take leaving the epiphyses of long bones at the adult knee joint. Here, we used transgenic mice that express fluorescent Venus protein in Schwann cells (Sox10-Venus mice) to visualize myelinated and unmyelinated nerves in the tibial epiphysis...
November 12, 2018: Bone
Selvaraj Vimalraj, Sivakamasundari Pichu, Thyagarajan Pankajam, Kasiviswanathan Dharanibalan, Valentin Djonov, Suvro Chatterjee
Angiogenesis is the formation of new blood vessels that occurs by two distinct processes following sprouting angiogenesis (SA) and intussusceptive angiogenesis (IA). Nitric oxide (NO) is known for its pro-angiogenic functions. However, no clear mechanisms are delineated on its role in promoting angiogenesis in reparative wound healing. We propose that NO regulates SA to IA transition and vice versa in wound milieu. We have used three models which include a new chick embryo extra-vasculature (CEV) burn wound model, adult Tie2-GFP transgenic Zebrafish caudal fin regeneration model and Zebrafish skin wound model to study the mechanisms underlying behind the role of NO in wound healing...
November 12, 2018: Nitric Oxide: Biology and Chemistry
Yuchen Xia, Martin Schlapschy, Volker Morath, Natalie Roeder, Elisabeth I Vogt, Daniela Stadler, Xiaoming Cheng, Ulf Dittmer, Kathrin Sutter, Mathias Heikenwalder, Arne Skerra, Ulrike Protzer
Interferon α (IFNα) so far is the only therapeutic option for chronic hepatitis B virus (HBV) infection that can lead to virus clearance. Unfortunately, its application is limited by side effects and response rates are low. The aim of this study was to generate a novel long-acting IFNα with the help of PASylation technology that adds a polypeptide comprising Proline, Alanine and Serine (PAS) to increase protein half-life. Following evaluation of four selected recombinant murine IFNα (mIFNα) subtypes in cell culture, the most active subtype mIFNα11 was fused with a 600 amino acid PAS-residue...
November 12, 2018: Antiviral Research
Bhupendra S Panwar, Jaswinder Kaur, Pradyumn Kumar, Sarvjeet Kaur
The novel cry52Ca1 gene from an Indian Bacillus thuringiensis (Bt) isolate was cloned in an expression vector (pET301/CT-DEST, 6xHis). The gene expressed as a ∼77.2 kDa protein in E. coli BL21-CodonPlus (DE3)-RIPL cells upon induction with isopropyl-thio-galactoside (IPTG) for 18 h at 28°C. Cry52Ca1 protein was toxic to Helicoverpa armigera (cotton bollworm) neonate larvae (LC50 36.66 µg per ml and MIC50 3.051 µg per ml) in diet-based laboratory assays. This gene has potential for deployment in insect-resistant transgenic crops...
November 12, 2018: Journal of Invertebrate Pathology
Jean-Luc Desseyn, Céline Portal, Frédéric Gottrand, Valérie Gouyer
Dysregulation of gel-forming mucins is associated with many airway diseases. Better knowledge of the pathophysiological mechanisms linking mucins and respiratory diseases will advance the understanding of their pathogenesis and should provide opportunities to develop new therapeutic compounds for treatment. MUC5B and MUC5AC are the two main gel-forming mucins in the respiratory tract. The organization in domains and the expression profile of mouse Muc5b are very similar to those in humans, which makes the mouse a relevant model for studies of the translational activities of human mucins...
November 2018: Annals of the American Thoracic Society
Haojie Huang, Jiaxiang Chen, Lisi Peng, Yao Yao, Defeng Deng, Yang Zhang, Yan Liu, Huamin Wang, Zhaoshen Li, Yan Bi, Ashley N Haddock, Xianbao Zhan, Weiqin Lu, Craig D Logsdon, Baoan Ji
INTRODUCTION: Replacement of the exocrine parenchyma by fibrous tissue is a main characteristic of chronic pancreatitis. Understanding the mechanisms of pancreatic fibrogenesis is critical for the development of preventive and therapeutic interventions. Cyclooxygenase-2 (COX-2), a rate-limiting enzyme for prostaglandin synthesis, is expressed in patients with chronic pancreatitis. However, it is unknown whether COX-2 can cause chronic pancreatitis. AIMS AND METHODS: To investigate the roles of pancreatic acinar COX-2 in fibrogenesis and the development of chronic pancreatitis, COX-2 was ectopically expressed specifically in pancreatic acinar cells in transgenic mice...
November 15, 2018: American Journal of Physiology. Gastrointestinal and Liver Physiology
Sunil Yadav, Katarzyna Kazmierczak, Jingsheng Liang, Yoel H Sitbon, Danuta Szczesna-Cordary
Myosin regulatory light chain (RLC) phosphorylation is important for cardiac muscle mechanics/function as well as for the Ca2+ -troponin/tropomyosin regulation of muscle contraction. This study focuses on the arginine to glutamine (R58Q) substitution in the human ventricular RLC (MYL2 gene), linked to malignant hypertrophic cardiomyopathy in humans and causing severe functional abnormalities in transgenic R58Q mice, including inhibition of cardiac RLC phosphorylation. Using a phosphomimic recombinant RLC variant where the phosphorylation site Ser-15 was substituted with an aspartic acid (S15D) and placed in the background of R58Q, we aimed to assess whether we could rescue/mitigate R58Q-induced structural/functional abnormalities in vitro...
November 15, 2018: FEBS Journal
Justin S Antony, Ngadhnjim Latifi, A K M Ashiqul Haque, Andrés Lamsfus-Calle, Alberto Daniel-Moreno, Sebastian Graeter, Praveen Baskaran, Petra Weinmann, Markus Mezger, Rupert Handgretinger, Michael S D Kormann
BACKGROUND: β-Thalassemia is an inherited hematological disorder caused by mutations in the human hemoglobin beta (HBB) gene that reduce or abrogate β-globin expression. Although lentiviral-mediated expression of β-globin and autologous transplantation is a promising therapeutic approach, the risk of insertional mutagenesis or low transgene expression is apparent. However, targeted gene correction of HBB mutations with programmable nucleases such as CRISPR/Cas9, TALENs, and ZFNs with non-viral repair templates ensures a higher safety profile and endogenous expression control...
November 14, 2018: Molecular and Cellular Pediatrics
Jennifer Wischhusen, Katheryne E Wilson, Jean-Guy Delcros, Rodolfo Molina-Peña, Benjamin Gibert, Shan Jiang, Jacqueline Ngo, David Goldschneider, Patrick Mehlen, Juergen K Willmann, Frederic Padilla
In ultrasound molecular imaging (USMI), ligand-functionalized microbubbles (MBs) are used to visualize vascular endothelial targets. Netrin-1 is upregulated in 60% of metastatic breast cancers and promotes tumor progression. A novel netrin-1 interference therapy requires the assessment of netrin-1 expression prior to treatment. In this study, we studied netrin-1 as a target for USMI and its potential as a companion diagnostic in breast cancer models. Methods: To verify netrin-1 expression and localization, an in vivo immuno-localization approach was applied, in which anti-netrin-1 antibody was injected into living mice 24 h before tumor collection, and revealed with secondary fluorescent antibody for immunofluorescence analysis...
2018: Theranostics
Matthias Brendel, Anna Jaworska, Felix Overhoff, Tanja Blume, Federico Probst, Franz-Josef Gildehaus, Peter Bartenstein, Christian Haass, Bernd Bohrmann, Jochen Herms, Michael Willem, Axel Rominger
Beta secretase (BACE) inhibitors are promising therapeutic compounds currently in clinical phase II/III trials. Preclinical [18 F]-florbetaben (FBB) amyloid PET imaging facilitates longitudinal monitoring of amyloidosis in Alzheimer's disease (AD) mouse models. Therefore, we applied this theranostic concept to investigate, by serial FBB PET, the efficacy of a novel BACE1 inhibitor in the PS2APP mouse, which is characterized by early and massive amyloid deposition. Methods: PS2APP and C57BL/6 (WT) mice were assigned to treatment (PS2APP: N=13; WT: N=11) and vehicle control (PS2APP: N=13; WT: N=11) groups at the age of 9...
2018: Theranostics
Kazuo Imaizumi, Yuchio Yanagawa, Guoping Feng, Charles C Lee
The thalamic reticular nucleus (TRN) is the main source of inhibition to the somatosensory thalamus (ventrobasal nucleus, VB) in mice. However, the functional topography and development of these projections with respect to the VB barreloids has been largely unexplored. In this respect, to assist in the study of these projections, we have utilized a vesicular gamma-aminobutryic acid (GABA) transporter (VGAT)-Venus transgenic mouse line to develop a brain slice preparation that enables the rapid identification of inhibitory neurons and projections...
2018: Frontiers in Neuroanatomy
Bo Sun, Aihong Zheng, Min Jiang, Shengling Xue, Qiao Yuan, Leiyu Jiang, Qing Chen, Mengyao Li, Yan Wang, Yong Zhang, Ya Luo, Xiaorong Wang, Fen Zhang, Haoru Tang
The clustered regulatory interspaced short palindromic repeat-associated protein 9 (CRISPR/Cas9) system has developed into a powerful gene-editing tool that has been successfully applied to various plant species. However, studies on the application of the CRISPR/Cas9 system to cultivated Brassica vegetables are limited. Here, we reported CRISPR/Cas9-mediated genome editing in Chinese kale (Brassica oleracea var. alboglabra) for the first time. A stretch of homologous genes, namely BaPDS1 and BaPDS2, was selected as the target site...
November 14, 2018: Scientific Reports
Luis Enrique Sandoval, Hongbing Jiang, David Wang
Orsay virus is the only known natural virus pathogen of Caenorhabditis elegans , and its discovery has enabled virus-host interactions studies in this model organism. Host genes required for viral infection remain understudied. We previously established a forward genetic screen based on a virus-inducible GFP transcriptional reporter to identify novel host factors essential for virus infection. Here we report the essential role in Orsay virus infection of the Dietary Restriction-Like ( drl-1 ) gene, which encodes a serine/threonine kinase similar to the mammalian MEKK3 kinase...
November 14, 2018: Journal of Virology
Zoltan Nagy, Timo Vögtle, Mitchell J Geer, Jun Mori, Silke Heising, Giada Di Nunzio, Ralph Gareus, Alexander Tarakhovsky, Arthur Weiss, Benjamin G Neel, Guillaume E Desanti, Alexandra Mazharian, Yotis A Senis
Conditional knockout (KO) mouse models are invaluable for elucidating the physiological roles of platelets. The Pf4-Cre transgenic mouse is the current model of choice for generating megakaryocyte/platelet-specific KO mice. Platelets and leukocytes work closely together in a wide range of disease settings, yet the specific contribution of platelets to these processes remains unclear. This is partially due to the Pf4-Cre transgene being expressed in a variety of leukocyte populations. To overcome this issue, we developed a Gp1ba-Cre transgenic mouse strain in which Cre expression in driven by the endogenous Gp1ba locus...
November 14, 2018: Blood
Catherine Fleitas, Gerard Piñol-Ripoll, Pau Marfull, Daniel Rocandio, Isidro Ferrer, Claire Rampon, Joaquim Egea, Carme Espinet
Alzheimer disease (AD) is a complex pathology related to multiple causes including oxidative stress. Brain-derived neurotrophic factor (BDNF) is a neutrotrophic factor essential for the survival and differentiation of neurons and is considered a key target in the pathophysiology of various neurodegenerative diseases, as for example AD. Contrarily to BDNF, the precursor form of BDNF (proBDNF) induces apoptosis through the specific interaction with p75 and its co-receptor, Sortilin.We used hippocampal tissue and cerebrospinal fluid from AD patients and controls...
November 14, 2018: Molecular Brain
Jia Xu, Ye Yuan, Ruining Zhang, Yanhui Song, Tianzhuo Sui, Jiaqi Wang, Chonghan Wang, Yujia Chen, Shuwen Guan, Liping Wang
Alzheimer's disease (AD) is a progressive neurodegenerative brain disease and is the most common cause of dementia in the elderly. The main hallmark of AD is the deposition of insoluble amyloid (Aβ) outside the neuron, leading to amyloid plaques and neurofibrillary tangles in the brain. Deuterohemin-Ala-His-Thr-Val-Glu-Lys (DhHP-6), a novel porphyrin-peptide, has both microperoxidase activity and cell permeability. In the present study, DhHP-6 efficiently inhibited the aggregation of Aβ and reduced the β-sheet percentage of Aβ from 89...
November 3, 2018: Bioorganic Chemistry
Jianbo Wu, Nadine Matthias, Jonathan Lo, Jose L Ortiz-Vitali, Annie W Shieh, Sidney H Wang, Radbod Darabi
Myogenic differentiation of human pluripotent stem cells (hPSCs) has been done by gene overexpression or directed differentiation. However, viral integration, long-term culture, and the presence of unwanted cells are the main obstacles. By using CRISPR/Cas9n, a double-reporter human embryonic stem cell (hESC) line was generated for PAX7/MYF5, allowing prospective readout. This strategy allowed pathway screen to define efficient myogenic induction in hPSCs. Next, surface marker screen allowed identification of CD10 and CD24 for purification of myogenic progenitors and exclusion of non-myogenic cells...
November 13, 2018: Cell Reports
Olha Nazarko, Amanuel Kibrom, Jana Winkler, Katherine Leon, Hannah Stoveken, Gabriel Salzman, Katarzyna Merdas, Yue Lu, Pradnya Narkhede, Gregory Tall, Simone Prömel, Demet Araç
Adhesion G-protein-coupled receptors (aGPCRs) play critical roles in diverse cellular processes in neurobiology, development, immunity, and numerous diseases. The lack of molecular understanding of their activation mechanisms, especially with regard to the transmembrane domains, hampers further studies to facilitate aGPCR-targeted drug development. Latrophilin-1/ADGRL1 is a model aGPCR that regulates synapse formation and embryogenesis, and its mutations are associated with cancer and attention-deficit/hyperactivity disorder...
May 25, 2018: iScience
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