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https://www.readbyqxmd.com/read/29777869/subconjunctival-dendrimer-drug-therapy-for-the-treatment-of-dry-eye-in-rabbits-with-induced-autoimmune-dacryoadenitis
#1
Hui Lin, Ying Liu, Siva P Kambhampati, Chih-Chien Hsu, Rangaramanujam M Kannan, Samuel C Yiu
PURPOSE: To investigate the efficacy of a single subconjunctival injection of dendrimer-dexamethasone to treat dry eye in a rabbit model of induced autoimmune dacryoadenitis (AID). METHOD: Dendrimer biodistribution after subconjuntival injection in AID animals was evaluated using Cy5-labelled dendrimer (D-Cy5) and confocal microscopy. Diseased animals were treated with free dexamethasone (Free-Dex), dendrimer-dexamethasone (D-Dex), or saline via a single subconjunctival injection...
May 16, 2018: Ocular Surface
https://www.readbyqxmd.com/read/29777339/characterization-of-a-cryptic-rolling-circle-replication-plasmid-pmk8-from-enterococcus-durans-1-8
#2
Penyu Zhou, Zhengyuan Zhai, Xiong Yao, Peihua Ma, Yanling Hao
A novel cryptic plasmid from Enterococcus durans 1-8, designated as pMK8, was sequenced and analyzed in this study. It consists of 3337 bp with a G + C content of 33.11%. Sequence analysis of pMK8 revealed three putative open reading frames (ORFs). Based on homology, two of them were identified as genes encoding replication initiation (RepC) and mobilization (Mob) protein, respectively. Sequence analysis revealed a pT181 family double-strand origin (dso) and a putative single-strand origin (sso) located upstream of the repC gene...
May 17, 2018: Current Microbiology
https://www.readbyqxmd.com/read/29773790/crispr-cas9-mediated-gene-targeting-in-arabidopsis-using-sequential-transformation
#3
Daisuke Miki, Wenxin Zhang, Wenjie Zeng, Zhengyan Feng, Jian-Kang Zhu
Homologous recombination-based gene targeting is a powerful tool for precise genome modification and has been widely used in organisms ranging from yeast to higher organisms such as Drosophila and mouse. However, gene targeting in higher plants, including the most widely used model plant Arabidopsis thaliana, remains challenging. Here we report a sequential transformation method for gene targeting in Arabidopsis. We find that parental lines expressing the bacterial endonuclease Cas9 from the egg cell- and early embryo-specific DD45 gene promoter can improve the frequency of single-guide RNA-targeted gene knock-ins and sequence replacements via homologous recombination at several endogenous sites in the Arabidopsis genome...
May 17, 2018: Nature Communications
https://www.readbyqxmd.com/read/29772841/the-effect-of-human-mesenchymal-stem-cells-derived-from-wharton-s-jelly-in-spinal-cord-injury-treatment-is-dose-dependent-and-can-be-facilitated-by-repeated-application
#4
Petr Krupa, Irena Vackova, Jiri Ruzicka, Kristyna Zaviskova, Jana Dubisova, Zuzana Koci, Karolina Turnovcova, Lucia Machova Urdzikova, Sarka Kubinova, Svatopluk Rehak, Pavla Jendelova
Human mesenchymal stem cells derived from Wharton's jelly (WJ-MSCs) were used for the treatment of the ischemic-compression model of spinal cord injury in rats. To assess the effectivity of the treatment, different dosages (0.5 or 1.5 million cells) and repeated applications were compared. Cells or saline were applied intrathecally by lumbar puncture for one week only, or in three consecutive weeks after injury. Rats were assessed for locomotor skills (BBB, rotarod, flat beam) for 9 weeks. Spinal cord tissue was morphometrically analyzed for axonal sprouting, sparing of gray and white matter and astrogliosis...
May 17, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29770951/requirement-of-tgf%C3%AE-signaling-for-effect-of-fluoride-on-osteoblastic-differentiation
#5
Jingmin Zhang, Ningning Jiang, Haolan Yu, Xiuhua Yu, Fengyang Guo, Zhitao Zhao, Hui Xu
Research focused on transforming growth factor β (TGFβ) signaling in osteoblast is gradually increasing, whereas literature is rare in terms of fluorosis. This work aimed to investigate how TGFβ signaling participated in regulation of the osteoblast by different doses of fluoride treatment. Bone marrow stem cells (BMSCs) were developed into osteoblastic cells and exposed to 1, 4, and 16 mg/L F- with and without 10 ng/mL of TGFβ. Cell viability and differentiation state of osteoblast under different settings were measured by means of cell counting kit and analysis of alkaline phosphatase (ALP) activity as well as formation of mineral nodules...
May 16, 2018: Biological Trace Element Research
https://www.readbyqxmd.com/read/29770349/single-step-precision-genome-editing-in-yeast-using-crispr-cas9
#6
Azat Akhmetov, Jon M Laurent, Jimmy Gollihar, Elizabeth C Gardner, Riddhiman K Garge, Andrew D Ellington, Aashiq H Kachroo, Edward M Marcotte
Genome modification in budding yeast has been extremely successful largely due to its highly efficient homology-directed DNA repair machinery. Several methods for modifying the yeast genome have previously been described, many of them involving at least two-steps: insertion of a selectable marker and substitution of that marker for the intended modification. Here, we describe a CRISPR-Cas9 mediated genome editing protocol for modifying any yeast gene of interest (either essential or nonessential) in a single-step transformation without any selectable marker...
March 20, 2018: Bio-protocol
https://www.readbyqxmd.com/read/29770147/koumine-attenuates-neuroglia-activation-and-inflammatory-response-to-neuropathic-pain
#7
Gui-Lin Jin, Sai-Di He, Shao-Mei Lin, Li-Mian Hong, Wan-Qing Chen, Ying Xu, Jian Yang, Su-Ping Li, Chang-Xi Yu
Despite decades of studies, the currently available drugs largely fail to control neuropathic pain. Koumine-an alkaloidal constituent derived from the medicinal plant Gelsemium elegans Benth.-has been shown to possess analgesic and anti-inflammatory properties; however, the underlying mechanisms remain unclear. In this study, we aimed to investigate the analgesic and anti-inflammatory effects and the possible underlying mechanisms of koumine. The analgesic and anti-inflammatory effects of koumine were explored by using chronic constriction injury of the sciatic nerve (CCI) neuropathic pain model in vivo and LPS-induced injury in microglia BV2 cells in vitro ...
2018: Neural Plasticity
https://www.readbyqxmd.com/read/29769698/mutation-screening-using-formalin-fixed-paraffin-embedded-tissues-a-stratified-approach-according-to-dna-quality
#8
Francesco Cucco, Alexandra Clipson, Hannah Kennedy, Joe Sneath Thompson, Ming Wang, Sharon Barrans, Moniek van Hoppe, Eguzkine Ochoa Ruiz, Josh Caddy, Debbie Hamid, Thomas Cummin, Cathy Burton, Andrew J Davies, Peter Johnson, Ming-Qing Du
DNA samples from formalin-fixed paraffin-embedded tissues are highly degraded with variable quality, and this imposes a big challenge for targeted sequencing due to false positives, largely caused by PCR errors and cytosine deamination. To eliminate false positives, a common practice is to validate the detected variants by Sanger sequencing or perform targeted sequencing in duplicate. Technically, PCR errors could be removed by molecular barcoding of template DNA prior to amplification as in the HaloPlexHS design...
May 16, 2018: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/29767364/single-cell-mrna-seq-using-the-fluidigm-c1-system-and-integrated-fluidics-circuits
#9
Haibiao Gong, Devin Do, Ramesh Ramakrishnan
Single-cell mRNA-seq is a valuable tool to dissect expression profiles and to understand the regulatory network of genes. Microfluidics is well suited for single-cell analysis owing both to the small volume of the reaction chambers and easiness of automation. Here we describe the workflow of single-cell mRNA-seq using C1 IFC, which can isolate and process up to 96 cells. Both on-chip procedure (lysis, reverse transcription, and preamplification PCR) and off-chip sequencing library preparation protocols are described...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29766378/2-methacryloyloxyethyl-phosphorylcholine-mpc-polymer-suppresses-an-increase-of-oral-bacteria-a-single-blind-crossover-clinical-trial
#10
Natsumi Fujiwara, Hiromichi Yumoto, Koji Miyamoto, Katsuhiko Hirota, Hiromi Nakae, Saya Tanaka, Keiji Murakami, Yasusei Kudo, Kazumi Ozaki, Yoichiro Miyake
OBJECTIVES: The biocompatible 2-methacryloyloxyethyl phosphorylcholine (MPC)-polymers, which mimic a biomembrane, reduce protein adsorption and bacterial adhesion and inhibit cell attachment. The aim of this study is to clarify whether MPC-polymer can suppress the bacterial adherence in oral cavity by a crossover design. We also investigated the number of Fusobacterium nucleatum, which is the key bacterium forming dental plaque, in clinical samples. MATERIALS AND METHODS: This study was a randomized, placebo-controlled, single-blind, crossover study, with two treatment periods separated by a 2-week washout period...
May 16, 2018: Clinical Oral Investigations
https://www.readbyqxmd.com/read/29765449/effects-of-single-and-combined-low-frequency-electromagnetic-fields-and-simulated-microgravity-on-gene-expression-of-human-mesenchymal-stem-cells-during-chondrogenesis
#11
Susanne Mayer-Wagner, Florian Hammerschmid, Helmut Blum, Stefan Krebs, Julia I Redeker, Boris M Holzapfel, Volkmar Jansson, Peter E Müller
Introduction: Low frequency electromagnetic fields (LF-EMF) and simulated microgravity (SMG) have been observed to affect chondrogenesis. A controlled bioreactor system was developed to apply LF-EMF and SMG singly or combined during chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in 3D culture. Material and methods: An external motor gear SMG bioreactor was combined with magnetic Helmholtz coils for EMF (5 mT; 15 Hz). Pellets of hMSCs (±TGF-β3) were cultured (P5) under SMG, LF-EMF, LF-EMF/SMG and control (1 g) conditions for 3 weeks...
April 2018: Archives of Medical Science: AMS
https://www.readbyqxmd.com/read/29764585/-construction-of-ezh2-knockout-animal-model-by-crispr-cas9-technology
#12
Fanrong Meng, Dan Zhao, Qinghua Zhou, Zhe Liu
BACKGROUND: It has been proven that CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated 9) system was the modern gene-editing technology through the constitutive expression of nucleases Cas9 in the mammalian, which binds to the specific site in the genome mediated by single-guide RNA (sgRNA) at desired genomic loci. The aim of this study is that the animal model of EZH2 gene knockout was constructed using CRISPR/Cas9 technology. METHODS: In this study, we designed two single-guide RNAs targeting the Exon3 and Exon4 of EZH2 gene...
May 20, 2018: Zhongguo Fei Ai za Zhi, Chinese Journal of Lung Cancer
https://www.readbyqxmd.com/read/29764204/biotechnological-potential-of-microbial-consortia-and-future-perspectives
#13
Shashi Kant Bhatia, Ravi Kant Bhatia, Yong-Keun Choi, Eunsung Kan, Yun-Gon Kim, Yung-Hun Yang
Design of a microbial consortium is a newly emerging field that enables researchers to extend the frontiers of biotechnology from a pure culture to mixed cultures. A microbial consortium enables microbes to use a broad range of carbon sources. It provides microbes with robustness in response to environmental stress factors. Microbes in a consortium can perform complex functions that are impossible for a single organism. With advancement of technology, it is now possible to understand microbial interaction mechanism and construct consortia...
May 15, 2018: Critical Reviews in Biotechnology
https://www.readbyqxmd.com/read/29761660/feasibility-of-re-biopsy-and-egfr-mutation-analysis-in-patients-with-non-small-cell-lung-cancer
#14
Tae-Ok Kim, In-Jae Oh, Bo Gun Kho, Ha Young Park, Jin Sun Chang, Cheol-Kyu Park, Hong-Joon Shin, Jung-Hwan Lim, Yong-Soo Kwon, Yu-Il Kim, Sung-Chul Lim, Young-Chul Kim, Yoo-Duk Choi
BACKGROUND: In cases of EGFR-tyrosine kinase inhibitor (TKI) failure, re-biopsy may be useful to understand resistance mechanisms and guide further treatment decisions. However, performing re-biopsy is challenging because of several hurdles. We assessed the feasibility of re-biopsy in advanced non-small cell lung cancer (NSCLC) patients in real-world clinical practice. METHODS: We retrospectively reviewed the clinical and pathologic data of advanced NSCLC patients who experienced disease progression after previous treatment with EGFR-TKIs at a single tertiary hospital in Korea between January 2014 and December 2016...
May 14, 2018: Thoracic Cancer
https://www.readbyqxmd.com/read/29754608/multiplex-polymerase-chain-reaction-in-combination-with-gel-electrophoresis-inductively-coupled-plasma-mass-spectrometry-a-powerful-tool-for-the-determination-of-gene-copy-number-variations-and-gene-expression-changes
#15
A Fernández Asensio, T Iglesias, A Cotarelo, M Espina, E Blanco-González, L M Sierra, M Montes-Bayón
During the last few years multiplex real-time or quantitative polymerase chain reaction PCR (qPCR) has become the method of choice for multiplex gene expression changes and gene copy number variations (CNVs) analysis. However, such determinations require the use of different fluorescent labels for the different amplified sequences, which increases significantly the costs of the analysis and limits the applicability of the technique for simultaneous amplification of many targets of interest in a single reaction...
September 6, 2018: Analytica Chimica Acta
https://www.readbyqxmd.com/read/29753810/dendrosomal-nanocurcumin-and-exogenous-p53-can-act-synergistically-to-elicit-anticancer-effects-on-breast-cancer-cells
#16
Narges Baghi, Babak Bakhshinejad, Reihaneh Keshavarz, Sadegh Babashah, Majid Sadeghizadeh
Triple-negative breast cancer (TNBC) constitutes an important histological subtype of breast cancer with a highly metastatic phenotype. The aim of the current study was to investigate the possible synergism between dendrosomal nanocurcumin (DNC) and exogenously delivered p53 in producing anticancer effects on a TNBC cell line. MTT assay was exploited to determine the viability of MDA-MB-231 cells against DNC and measure the impact of p53 overexpresssion on DNC-related cytotoxicity. Annexin-V/PI staining followed by flow cytometry and wound healing assay were used to evaluate the effects of DNC and exogenous p53, alone and in combination, on apoptosis induction and migratory capacity of MDA-MB-231 cells, respectively...
May 10, 2018: Gene
https://www.readbyqxmd.com/read/29752127/implementation-of-next-generation-sequencing-technology-for-somatic-mutation-detection-in-routine-laboratory-practice
#17
Tindaro Giardina, Cleo Robinson, Fabienne Grieu-Iacopetta, Michael Millward, Barry Iacopetta, Dominic Spagnolo, Benhur Amanuel
The introduction of next generation sequencing (NGS) in the routine diagnostic setting is still in the development phase and has been limited by its complexity. Targeted NGS offers an attractive alternative to performing multiple single target assays and is very useful in meeting the increasing clinical demand for testing of multiple genetic aberrations in cancer specimens. To this end, we carried out a blinded validation study on 113 tumours in a diagnostic laboratory and compared mutation results from targeted NGS with those from Sanger sequencing, pyrosequencing, competitive allele specific TaqMan polymerase chain reaction (CAST PCR) and Cobas assays...
May 8, 2018: Pathology
https://www.readbyqxmd.com/read/29751831/intravenous-injection-of-allogeneic-umbilical-cord-derived-multipotent-mesenchymal-stromal-cells-reduces-the-infarct-area-and-ameliorates-cardiac-function-in-a-porcine-model-of-acute-myocardial-infarction
#18
Meikuang Lim, Weiqiang Wang, Lu Liang, Zhi-Bo Han, Zongjin Li, Jie Geng, Meng Zhao, Honghong Jia, Jie Feng, Zhe Wei, Baoquan Song, Jiemin Zhang, Jun Li, Tianwen Liu, Fan Wang, Ting Li, Jianming Li, Yihu Fang, Jianhua Gao, Zhongchao Han
BACKGROUND: Multipotent mesenchymal stromal cell (MSC) therapy has been widely recognized as a feasible strategy for regenerating injured myocardial tissue. However, little is known about the efficacy of intravenous injection of allogeneic umbilical cord (UC) MSCs in preclinical models of porcine myocardial infarction. METHODS: Different dosages of allogeneic UC-MSCs or the vehicle [phosphate-buffered saline (PBS)] were delivered intravenously into an acute myocardial infarction (AMI) porcine model twice after coronary ligation...
May 11, 2018: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29751033/a-novel-caspase-associated-recruitment-domain-card-containing-protein-cgcardcp-1-involved-in-lps-recognition-and-nf-%C3%AE%C2%BAb-activation-in-oyster-crassostrea-gigas
#19
Feifei Wang, Zichao Yu, Weilin Wang, Yiqun Li, Guangxia Lu, Chen Qu, Hui Wang, Mengmeng Lu, Lingling Wang, Linsheng Song
Caspase-associated recruitment domain (CARD) containing proteins play critical roles in molecular interaction and regulation of various signaling pathways, such as the activation of caspase and NF-κB singling pathway in the process of apoptosis or inflammation. In the present study, a novel CARD containing protein (designed CgCARDCP-1) was identified and characterized from oyster Crassostrea gigas. Molecular feature analysis revealed that, the open reading frame (ORF) of CgCARDCP-1 gene was 759 bp encoding a polypeptide of 253 amino acids with a conserved N-terminal CARD domain and two transcriptional coactivator p15 (PC4) domains in C-terminus...
May 8, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29749487/rs2910164-in-microrna%C3%A2-146a-confers-an-elevated-risk-of-depression-in-patients-with-coronary-artery-disease-by-modulating-the-expression-of-nos1
#20
Xinling Zhang, Qianqian Huo, Wei Sun, Chunxiang Zhang, Zongyin Wu, Bing Xing, Qiang Li
Depression has been well established as an independent predictor of mortality and cardiac morbidity rates in patients with coronary artery disease (CAD). Evidence has shown that single nucleotide polymorphisms located in pre‑microRNA (miRNA) or mature miRNA may modify various biological processes and affect the process of carcinogenesis, and the downregulation of neuronal nitric oxide synthase 1 (NOS1) can induce depression. It has been shown that NOS1 is the target gene of miR‑146a, and that the rs2910164 G/C polymorphism can downregulate the expression of miR‑146a...
April 26, 2018: Molecular Medicine Reports
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