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https://www.readbyqxmd.com/read/28624224/optimizing-the-dna-donor-template-for-homology-directed-repair-of-double-strand-breaks
#1
Fei Song, Knut Stieger
The CRISPR-Cas (clustered regularly interspaced short palindromic repeats-associated proteins) technology enables rapid and precise genome editing at any desired genomic position in almost all cells and organisms. In this study, we analyzed the impact of different repair templates on the frequency of homology-directed repair (HDR) and non-homologous end joining (NHEJ). We used a stable HEK293 cell line expressing the traffic light reporter (TLR-3) system to quantify HDR and NHEJ events following transfection with Cas9, eight different guide RNAs, and a 1,000 bp donor template generated either as circular plasmid, as linearized plasmid with long 3' or 5' backbone overhang, or as PCR product...
June 16, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28620261/single-nucleotide-polymorphism-based-genetic-diversity-analysis-of-the-kilakarsal-and-vembur-sheep-breeds
#2
Rathinasamy Selvam, Nagarajan Murali, A Kannan Thiruvenkadan, Ramesh Saravanakumar, Gurusamy Ponnudurai, Thilak Pon Jawahar
AIM: The present study was thus undertaken to analyze the genetic diversity of Kilakarsal and Vembur sheep breeds using single-nucleotide polymorphism (SNP) markers within Toll-like receptor (TLR) 3, 5, 6, 9, and 10 genes. MATERIALS AND METHODS: Competitive allele-specific polymerase chain reaction (PCR)-based end-point genotyping was performed using real-time PCR to type the SNPs. Allele discrimination module implemented in real-time PCR was utilized to call the genotypes based on fluorescence intensity recorded for each of the two alleles...
May 2017: Veterinary World
https://www.readbyqxmd.com/read/28613415/composition-and-functionality-of-the-intrahepatic-innate-lymphoid-cell-compartment-in-human-non-fibrotic-and-fibrotic-livers
#3
Marianne Forkel, Lena Berglin, Eliisa Kekäläinen, Adrian Carlsson, Emma Svedin, Jakob Michaëlsson, Maho Nagasawa, Jonas S Erjefält, Michiko Mori, Malin Flodström-Tullberg, Annika Bergquist, Hans-Gustaf Ljunggren, Magnus Westgren, Ulrik Lindforss, Danielle Friberg, Carl Jorns, Ewa Ellis, Niklas K Björkström, Jenny Mjösberg
Human innate lymphoid cells have been described to exist in different organs, with functional deregulation of these cells contributing to several disease states. Here, we performed the first detailed characterization of the phenotype, tissue-residency properties and functionality of ILC1s, ILC2s and ILC3s in the human adult and fetal liver. In addition, we investigated changes in the ILC compartment in liver fibrosis. A unique composition of tissue-resident ILCs was observed in non-fibrotic livers as compared with that in mucosal tissues, with NKp44(-) ILC3s accounting for the majority of total intrahepatic ILCs...
June 14, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28612711/alleviative-effects-from-boswellic-acid-on-acetaminophen-induced-hepatic-injury-corrected-and-republished-from-biomedicine-taipei-2016-jun-6-2-9-doi-10-7603-s40681-016-0009-1pmcid-pmc4864770
#4
Lung-Che Chen, Li-Hong Hu, Mei-Chin Yin
Protective effects of boswellic acid (BA) against acetaminophen (APAP)-induced hepatotoxicity in Balb/ cA mice were examined. BA, at 0.05 or 0.1%, was supplied for 4 weeks. Acute liver injury was induced by APAP treatment. Results showed that BA intake increased hepatic BA bioavailability. APAP treatment decreased glutathione (GSH) level, increased reactive oxygen species (ROS) and oxidized glutathione (GSSG) production; and lowered activity and protein expression of glutathione reductase (GR) and heme oxygenase (HO)-1 in liver...
June 2017: BioMedicine
https://www.readbyqxmd.com/read/28592438/cd14-cd15-hla-dr-myeloid-derived-suppressor-cells-impair-antimicrobial-responses-in-patients-with-acute-on-chronic-liver-failure
#5
Christine Bernsmeier, Evangelos Triantafyllou, Robert Brenig, Fanny J Lebosse, Arjuna Singanayagam, Vishal C Patel, Oltin T Pop, Wafa Khamri, Rooshi Nathwani, Robert Tidswell, Christopher J Weston, David H Adams, Mark R Thursz, Julia A Wendon, Charalambos Gustav Antoniades
OBJECTIVE: Immune paresis in patients with acute-on-chronic liver failure (ACLF) accounts for infection susceptibility and increased mortality. Immunosuppressive mononuclear CD14(+)HLA-DR(-) myeloid-derived suppressor cells (M-MDSCs) have recently been identified to quell antimicrobial responses in immune-mediated diseases. We sought to delineate the function and derivation of M-MDSC in patients with ACLF, and explore potential targets to augment antimicrobial responses. DESIGN: Patients with ACLF (n=41) were compared with healthy subjects (n=25) and patients with cirrhosis (n=22) or acute liver failure (n=30)...
June 7, 2017: Gut
https://www.readbyqxmd.com/read/28539706/multiple-immunosuppressive-effects-of-cpg-c41-on-intracellular-tlr-mediated-inflammation
#6
Wancheng Liu, Xuejiao Yang, Ning Wang, Shijun Fan, Yuanfeng Zhu, Xinchuan Zheng, Yan Li
A growing body of literature suggests that most chronic autoimmune diseases are associated with inappropriate inflammation mediated by Toll-like receptor (TLR) 3, TLR7/8, or TLR9. Therefore, research into blocking TLR activation to treat these disorders has become a hot topic. Here, we report the immunomodulatory properties of a nonstimulatory CpG-containing oligodeoxynucleotide (CpG-ODN), CpG-c41, which had previously only been known as a TLR9 antagonist. In this study, we found that both in vitro and in vivo CpG-c41 decreased levels of various proinflammatory factors that were induced by single activation or coactivation of intracellular TLRs, but not membrane-bound TLRs, no matter what downstream signal pathways the TLRs depend on...
2017: Mediators of Inflammation
https://www.readbyqxmd.com/read/28448526/human-alkaline-phosphatase-dephosphorylates-microbial-products-and-is-elevated-in-preterm-neonates-with-a-history-of-late-onset-sepsis
#7
Matthew Pettengill, Juan D Matute, Megan Tresenriter, Julie Hibbert, David Burgner, Peter Richmond, José Luis Millán, Al Ozonoff, Tobias Strunk, Andrew Currie, Ofer Levy
BACKGROUND: A host defense function for Alkaline phosphatases (ALPs) is suggested by the contribution of intestinal ALP to detoxifying bacterial lipopolysaccharide (endotoxin) in animal models in vivo and the elevation of ALP activity following treatment of human cells with inflammatory stimuli in vitro. However the activity of ALP in human plasma (primarily tissue-nonspecific ALP; TNAP) on lipopolysaccharide and other microbial products has not been assessed, nor has its expression been studied in preterm newborns, a vulnerable population at high risk of sepsis...
2017: PloS One
https://www.readbyqxmd.com/read/28422714/toll-like-receptor-3-as-an-immunotherapeutic-target-for-kras-mutated-colorectal-cancer
#8
Radhashree Maitra, Titto Augustine, Yitzchak Dayan, Carol Chandy, Matthew Coffey, Sanjay Goel
New therapeutic interventions are essential for improved management of patients with metastatic colorectal cancer (mCRC). This is especially critical for those patients whose tumors harbor a mutation in the KRAS oncogene (40-45% of all patients). This patient cohort is excluded from receiving anti-EGFR monoclonal antibodies that have added a significant therapeutic benefit for KRAS wild type CRC patients. Reovirus, a double stranded (ds) RNA virus is in clinical development for patients with chemotherapy refractory KRAS mutated tumors...
May 23, 2017: Oncotarget
https://www.readbyqxmd.com/read/28402971/implicating-receptor-activator-of-nf-%C3%AE%C2%BAb-rank-rank-ligand-signalling-in-microglial-responses-to-toll-like-receptor-stimuli
#9
Anton Kichev, Pascale Eede, Pierre Gressens, Claire Thornton, Henrik Hagberg
Inflammation in the perinatal brain caused by maternal or intrauterine fetal infection is now well established as an important contributor to the development of perinatal brain injury. Exposure to inflammatory products can impair perinatal brain development and act as a risk factor for neurological dysfunction, cognitive disorders, cerebral palsy, or preterm birth. Pre-exposure to inflammation significantly exacerbates brain injury caused by hypoxic/ischaemic insult. Tumour necrosis factor (TNF) is a family of cytokines largely involved in inflammation signalling...
April 13, 2017: Developmental Neuroscience
https://www.readbyqxmd.com/read/28388966/vaccination-with-poly-ic-lc-and-peptide-pulsed-autologous-dendritic-cells-in-patients-with-pancreatic-cancer
#10
Shikhar Mehrotra, Carolyn D Britten, Steve Chin, Elizabeth Garrett-Mayer, Colleen A Cloud, Mingli Li, Gina Scurti, Mohamed L Salem, Michelle H Nelson, Melanie B Thomas, Chrystal M Paulos, Andres M Salazar, Michael I Nishimura, Mark P Rubinstein, Zihai Li, David J Cole
BACKGROUND: Dendritic cells (DCs) enhance the quality of anti-tumor immune response in patients with cancer. Thus, we posit that DC-based immunotherapy, in conjunction with toll-like receptor (TLR)-3 agonist poly-ICLC, is a promising approach for harnessing immunity against metastatic or locally advanced unresectable pancreatic cancer (PC). METHODS: We generated autologous DCs from the peripheral blood of HLA-A2(+) patients with PC. DCs were pulsed with three distinct A2-restricted peptides: 1) human telomerase reverse transcriptase (hTERT, TERT572Y), 2) carcinoembryonic antigen (CEA; Cap1-6D), and 3) survivin (SRV...
April 7, 2017: Journal of Hematology & Oncology
https://www.readbyqxmd.com/read/28273893/dengue-virus-infection-increases-microglial-cell-migration
#11
Ming-Kai Jhan, Tsung-Ting Tsai, Chia-Ling Chen, Cheng-Chieh Tsai, Yi-Lin Cheng, Yi-Chao Lee, Chiung-Yuan Ko, Yee-Shin Lin, Chih-Peng Chang, Liang-Tzung Lin, Chiou-Feng Lin
Activated microglial cells are present in dengue virus (DENV)-infected brains; however, the possible effects of DENV on microglia remain unclear. Here, we demonstrated DENV caused infection, including viral entry, RNA replication, viral protein expression, and virus release, in the murine microglial cell line BV2. DENV infection caused an increase in the formation of the multipolar phenotype in vitro and in vivo without affecting cell growth and cytotoxicity. DENV infection considerably increased cell motility and disrupting either actin filaments or clathrin retarded such effect...
December 2017: Scientific Reports
https://www.readbyqxmd.com/read/28266028/triggering-of-tlr-3-4-nod2-and-dc-sign-reduces-viral-replication-and-increases-t-cell-activation-capacity-of-hiv-infected-human-dendritic-cells
#12
Sylvain Cardinaud, Alejandra Urrutia, Angeline Rouers, Pierre-Grégoire Coulon, Jérome Kervevan, Clémence Richetta, Anne Bet, Emmanuel A Maze, Martin Larsen, Maria-Candela Iglesias, Victor Appay, Stéphanie Graff-Dubois, Arnaud Moris
A variety of signals influence the capacity of dendritic cells (DCs) to mount potent antiviral cytotoxic T-cell (CTL) responses. In particular, innate immune sensing by pathogen recognition receptors, such as TLR and C-type lectines, influences DC biology and affects their susceptibility to HIV infection. Yet, whether the combined effects of PPRs triggering and HIV infection influence HIV-specific (HS) CTL responses remain enigmatic. Here, we dissect the impact of innate immune sensing by pathogen recognition receptors on DC maturation, HIV infection, and on the quality of HS CTL activation...
May 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28262465/characterization-of-human-fibroblastic-reticular-cells-as-potential-immunotherapeutic-tools
#13
Jaris Valencia, Eva Jiménez, Víctor G Martínez, Beatriz G Del Amo, Laura Hidalgo, Ana Entrena, Lidia M Fernández-Sevilla, Francisco Del Río, Alberto Varas, Ángeles Vicente, Rosa Sacedón
Fibroblastic reticular cells (FRCs) are essential players during adaptive immune responses not only as a structural support for the encounter of antigen-presenting cells and naive T lymphocytes but also as a source of modulatory signals. However, little is known about this cell population in humans. To address the phenotypical and functional analysis of human FRCs here we established splenic (SP) and mesenteric lymph node (LN) CD45(-)CD31(-)CD90(+)podoplanin(+) myofibroblastic cell cultures. They shared the phenotypical characteristics distinctive of FRCs, including the expression of immunomodulatory factors and peripheral tissue antigens...
May 2017: Cytotherapy
https://www.readbyqxmd.com/read/28258480/the-anti-hsv-1-effect-of-quercetin-is-dependent-on-the-suppression-of-tlr-3-in-raw-264-7-cells
#14
Seulki Lee, Hwan Hee Lee, Yu Su Shin, Hyojeung Kang, Hyosun Cho
Quercetin is a major component of the plant Glycyrrhiza uralensis, which is largely used as a traditional medicine in Asia. Quercetin has been reported to have several biological activities, which include anti-viral and anti-inflammatory effects. We explored the molecular mechanism linking anti-viral and anti-inflammatory activities using an in vitro herpes simplex virus-1 (HSV-1) infection model. Raw 264.7 cells were infected with HSV-1 in the presence or absence of different concentrations of quercetin and infected cell lysates were harvested 24 h later...
March 3, 2017: Archives of Pharmacal Research
https://www.readbyqxmd.com/read/28251511/herpes-simplex-encephalitis-an-update
#15
REVIEW
John W Gnann, Richard J Whitley
PURPOSE OF REVIEW: The goal of this review is to provide an update on current thinking regarding herpes simplex encephalitis (HSE), emphasizing new information about pathogenesis, diagnosis, and immune responses. Specific questions to be addressed are the following: (1) Is there a genetic predisposition to HSE? (2) What clinical approaches have the greatest impact on improving the long-term outcomes in patients with HSE? And (3) are there immune-mediated mechanisms that may account for relapsing HSE? RECENT FINDINGS: Toll-like receptor 3 (TLR 3) plays an important role in innate immune responses, including generation of interferons...
March 2017: Current Infectious Disease Reports
https://www.readbyqxmd.com/read/28222448/acute-effects-of-viral-exposure-on-p-glycoprotein-function-in-the-mouse-fetal-blood-brain-barrier
#16
Enrrico Bloise, Sophie Petropoulos, Majid Iqbal, Alisa Kostaki, Tania Maria Ortiga-Carvalho, William Gibb, Stephen G Matthews
BACKGROUND/AIMS: Viral infection during pregnancy is known to affect the fetal brain. The toll-like receptor (TLR)-3 is a pattern recognition receptor activated by viruses known to elicit adverse fetal neurological outcomes. The P-glycoprotein (P-gp) efflux transporter protects the developing fetus by limiting the transfer of substrates across both the placenta and the fetal blood-brain barrier (BBB). As such, inhibition of P-gp at these blood-barrier sites may result in increased exposure of the developing fetus to environmental toxins and xenobiotics present in the maternal circulation...
2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28210837/toll-like-receptor-3-tlr-3-tlr-4-and-cd80-expression-in-peripheral-blood-mononuclear-cells-and-urinary-cd80-levels-in-children-with-idiopathic-nephrotic-syndrome
#17
Om P Mishra, Ravindra Kumar, Gopeshwar Narayan, Pradeep Srivastava, Abhishek Abhinay, Rajniti Prasad, Ankur Singh, Vineeta V Batra
BACKGROUND: The aims of this study were (1) to detect toll-like receptor (TLR)-3, TLR-4 and CD80 expression in peripheral blood mononuclear cells (PBMCs) and estimate urinary CD80 levels in children with idiopathic nephrotic syndrome and (2) to investigate the utility of these markers to differentiate between biopsy-proven minimal change disease (MCD) and focal segmental glomeruloscelerosis (FSGS). METHODS: The study included 70 patients with idiopathic nephrotic syndrome (NS), of whom 40 had steroid-sensitive NS (SSNS; 25 with active NS, 15 in remission) and 30 had steroid-resistant NS (SRNS) patients, and 23 healthy controls...
February 16, 2017: Pediatric Nephrology: Journal of the International Pediatric Nephrology Association
https://www.readbyqxmd.com/read/28192646/higher-expression-of-toll-like-receptors-3-7-8-and-9-in-pityriasis-rosea
#18
Mostafa Abou El-Ela, Mohamed El-Komy, Rania Abdel Hay, Rehab Hegazy, Amin Sharobim, Laila Rashed, Khalda Amr
BACKGROUND: Pityriasis rosea (PR) is a common papulosquamous skin disease in which an infective agent may be implicated. Toll-like receptors (TLRs) play an important role in immune responses and in the pathophysiology of inflammatory skin diseases. Our aim was to determine the possible roles of TLRs 3, 7, 8, and 9 in the pathogenesis of PR. METHODS: Twenty-four PR patients and 24 healthy individuals (as controls) were included in this case control study. All recruits were subjected to routine laboratory investigations...
March 2017: Journal of Pathology and Translational Medicine
https://www.readbyqxmd.com/read/28138030/the-tlr3-agonist-inhibit-drug-efflux-and-sequentially-consolidates-low-dose-cisplatin-based-chemoimmunotherapy-while-reducing-side-effects
#19
Liang Ding, Jing Ren, Dongya Zhang, Yi Li, Xiaofeng Huang, Jianjian Ji, Qingang Hu, Hui Wang, Yanhong Ni, Yayi Hou
The traditional maximum dose density chemotherapy renders the tumor patients not only the tumor remission but the chemotherapy resistance and more adverse side effects. According to the widely positive expression of Toll-like receptor (TLR)-3 in oral squamous cell carcinoma (OSCC) patients (n = 166), we here provided an alternative strategy involved the orderly treatment of TLR3 agonist polyinosine-polycytidylic acid (PIC) and low-dose cisplatin. The optimal dose of cisplatin, the novel role of PIC and the side effects of the combined chemotherapy were determined in vitro and in distinct human tumor models in vivo The results in vitro indicated that preculture with PIC downregulated drug transporters (e...
June 2017: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/28130499/amino-acid-catabolism-in-multiple-sclerosis-affects-immune-homeostasis
#20
Laura Negrotto, Jorge Correale
Amino acid catabolism has been implicated in immunoregulatory mechanisms present in several diseases, including autoimmune disorders. Our aims were to assess expression and activity of enzymes involved in Trp and Arg catabolism, as well as to investigate amino acid catabolism effects on the immune system of multiple sclerosis (MS) patients. To this end, 40 MS patients, 30 healthy control subjects, and 30 patients with other inflammatory neurological diseases were studied. Expression and activity of enzymes involved in Trp and Arg catabolism (IDO1, IDO2, Trp 2,3-dioxygenase [TDO], arginase [ARG] 1, ARG2, inducible NO synthetase) were evaluated in PBMCs...
March 1, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
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