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https://www.readbyqxmd.com/read/28212413/scarless-deletion-of-up-to-seven-methyl-accepting-chemotaxis-genes-with-an-optimized-method-highlights-key-function-of-chem-in-salmonella-typhimurium
#1
Stefanie Hoffmann, Christiane Schmidt, Steffi Walter, Jennifer K Bender, Roman G Gerlach
Site-directed scarless mutagenesis is an essential tool of modern pathogenesis research. We describe an optimized two-step protocol for genome editing in Salmonella enterica serovar Typhimurium to enable multiple sequential mutagenesis steps in a single strain. The system is based on the λ Red recombinase-catalyzed integration of a selectable antibiotics resistance marker followed by replacement of this cassette. Markerless mutants are selected by expressing the meganuclease I-SceI which induces double-strand breaks in bacteria still harboring the resistance locus...
2017: PloS One
https://www.readbyqxmd.com/read/28211068/dentine-matrix-proteins-isolation-and-effects-on-human-pulp-cells
#2
M Widbiller, A Eidt, S R Lindner, K-A Hiller, H Schweikl, W Buchalla, K M Galler
AIM: To establish a simplified and efficient protocol for the isolation and concentration of matrix proteins from human dentine, and to assess the effects of extracted dentine matrix proteins (eDMP) on the behavior of human pulp cells. METHODOLOGY: Matrix proteins were isolated from human dentine, purified, concentrated and characterized with protein and enzyme-linked immunosorbent assays. Culture media were supplemented with eDMP in different concentrations, referred to as eDMP 1 to 10,000, to assess viability and proliferation of human pulp cells by DNA and MTT assays; apoptotic events were quantified by flow cytometry...
February 17, 2017: International Endodontic Journal
https://www.readbyqxmd.com/read/28210968/study-on-critical-sized-ultra-high-molecular-weight-polyethylene-wear-particles-loaded-with-alendronate-sodium-in-vitro-release-and-cell-response
#3
Yumei Liu, Feng Shi, Kemeng Gong, Yang Liu, Wei Zhi, Jie Weng, Shuxin Qu
The aim of this study was to investigate the in vitro release and the effect of RAW 264.7 macrophages of critical-sized wear particles of ultra-high molecular weight polyethylene (UHMWPE) loaded with alendronate sodium (ALN), one of the most effective drugs to treat osteoporosis in clinic. The critical-sized UHMWPE-ALN 0.5 wt.% wear particles were prepared by vacuum gradient filtration combined with Pluronic F-68. In vitro release of ALN from critical-sized UHMWPE-ALN wear particles was investigated in phosphate buffered saline (PBS) at 37 °C with a shaker...
April 2017: Journal of Materials Science. Materials in Medicine
https://www.readbyqxmd.com/read/28209513/isoamyl-alcohol-odor-promotes-longevity-and-stress-tolerance-via-daf-16-in-caenorhabditis-elegans
#4
Chiho Kurino, Tsubasa Furuhashi, Kaori Sudoh, Kazuichi Sakamoto
The possibility that odor plays a role in lifespan regulation through effects on the nervous system is indicated by research on Caenorhabditis elegans. In fact, ablation of AWA and AWC, which are suggested as olfactory neurons, has been shown to extend lifespan via DAF-16, a homolog of FoxO. However, the effects of odor stimuli on the lifespan still remain unclear. Thus, we here aimed to clarify the effect of attractive and repulsive odors on longevity and stress tolerance in C. elegans and to analyze the pathways thereof...
February 13, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28208423/modeling-angiogenesis-a-discrete-to-continuum-description
#5
Samara Pillay, Helen M Byrne, Philip K Maini
Angiogenesis is the process by which new blood vessels develop from existing vasculature. During angiogenesis, endothelial tip cells migrate via diffusion and chemotaxis, loops form via tip-to-tip and tip-to-sprout anastomosis, new tip cells are produced via branching, and a vessel network forms as endothelial cells follow the paths of tip cells. The latter process is known as the snail trail. We use a mean-field approximation to systematically derive a continuum model from a two-dimensional lattice-based cellular automaton model of angiogenesis in the corneal assay, based on the snail-trail process...
January 2017: Physical Review. E
https://www.readbyqxmd.com/read/28207738/testing-the-limits-of-gradient-sensing
#6
Vinal Lakhani, Timothy C Elston
The ability to detect a chemical gradient is fundamental to many cellular processes. In multicellular organisms gradient sensing plays an important role in many physiological processes such as wound healing and development. Unicellular organisms use gradient sensing to move (chemotaxis) or grow (chemotropism) towards a favorable environment. Some cells are capable of detecting extremely shallow gradients, even in the presence of significant molecular-level noise. For example, yeast have been reported to detect pheromone gradients as shallow as 0...
February 16, 2017: PLoS Computational Biology
https://www.readbyqxmd.com/read/28207159/the-critical-role-of-the-znf217-oncogene-in-promoting-breast-cancer-metastasis-to-the-bone
#7
Aurélie Bellanger, Caterina F Donini, Julie A Vendrell, Jonathan Lavaud, Irma Machuca-Gayet, Maëva Ruel, Julien Vollaire, Evelyne Grisard, Balázs Győrffy, Ivan Bièche, Olivier Peyruchaud, Jean-Luc Coll, Isabelle Treilleux, Véronique Maguer-Satta, Véronique Josserand, Pascale A Cohen
Bone metastasis affects more than 70% of patients with advanced breast cancer. However, the molecular mechanisms underlying this process remain unclear. Based on the analysis of clinical datasets, and in vitro and in vivo experiments, we report that the ZNF217 oncogene is a crucial mediator and indicator of bone metastasis. Patients with high ZNF217 mRNA expression levels in primary breast tumours had a higher risk of developing bone metastases. MDA-MB-231 breast cancer cells stably transfected with ZNF217 (MDA-MB-231-ZNF217) displayed the dysregulated expression of a set of genes with bone homing and metastasis characteristics, which overlapped with two previously described "osteolytic bone metastasis" gene signatures, while also highlighting the bone morphogenetic protein (BMP) pathway...
February 16, 2017: Journal of Pathology
https://www.readbyqxmd.com/read/28205317/oxidative-stress-and-immunosenescence-in-spleen-of-obese-mice-can-be-reversed-by-2-hydroxyoleic-acid
#8
Alina Gheorghe, Fátima Pérez de Heredia, Caroline Hunsche, Noemí Redondo, Ligia Esperanza Díaz, Oskarina Hernández, Ascensión Marcos, Mónica De la Fuente
We aimed to investigate the effects of obesity on oxidative stress and leukocyte function in spleen of mice, and to assess whether supplementation with 2-hydroxyoleic acid (2-OHOA) or n-3 polyunsaturated fatty acids (PUFA) could reverse those effects. Female ICR/CD1 mice (8 weeks old, n = 24) received an obesogenic diet (22% fat for 4 weeks and 60% fat for 14 weeks). After 6 weeks, mice were split in three groups (n = 8/group): no supplementation, 2-OHOA supplementation (1500 mg kg(-1) ) and n-3 PUFA supplementation (EPA + DHA, 3000 mg kg(-1) diet)...
February 15, 2017: Experimental Physiology
https://www.readbyqxmd.com/read/28202513/myeloid-cells-that-impair-immunotherapy-are-restored-in-melanomas-which-acquire-resistance-to-braf-inhibitors
#9
Shannon M Steinberg, Tamer Shabaneh, Peisheng Zhang, Viktor Martyanov, Zhenghui Li, Brian Malik, Tammara Wood, Andrea Boni, Aleksey Molodtsov, Christina V Angeles, Tyler J Curiel, Michael Whitfield, Mary Jo Turk
Acquired resistance to BRAFV600E inhibitors (BRAFi) in melanoma remains a common clinical obstacle, as is the case for any targeted drug therapy that can be developed given the plastic nature of cancers. While there has been significant focus on the cancer cell-intrinsic properties of BRAFi resistance, the impact of BRAFi resistance on host immunity has not been explored. Here we provide preclinical evidence that resistance to BRAFi in an autochthonous mouse model of melanoma is associated with restoration of myeloid-derived suppressor cells (MDSC) in the tumor microenvironment initially reduced by BRAFi treatment...
February 15, 2017: Cancer Research
https://www.readbyqxmd.com/read/28197202/expression-changes-of-nerve-cell-adhesion-molecules-l1-and-semaphorin-3a-after-peripheral-nerve-injury
#10
Qian-Ru He, Meng Cong, Qing-Zhong Chen, Ya-Feng Sheng, Jian Li, Qi Zhang, Fei Ding, Yan-Pei Gong
The expression of nerve cell adhesion molecule L1 in the neuronal growth cone of the central nervous system is strongly associated with the direction of growth of the axon, but its role in the regeneration of the peripheral nerve is still unknown. This study explored the problem in a femoral nerve section model in rats. L1 and semaphorin 3A mRNA and protein expressions were measured over the 4-week recovery period. Quantitative polymerase chain reaction showed that nerve cell adhesion molecule L1 expression was higher in the sensory nerves than in motor nerves at 2 weeks after injury, but vice versa for the expression of semaphorin 3A...
December 2016: Neural Regeneration Research
https://www.readbyqxmd.com/read/28194725/otophylloside-b-protects-against-a%C3%AE-toxicity-in-caenorhabditis-elegans-models-of-alzheimer-s-disease
#11
Jie Yang, Xiao-Bing Huang, Qin-Li Wan, Ai-Jun Ding, Zhong-Lin Yang, Ming-Hua Qiu, Hua-Ying Sun, Shu-Hua Qi, Huai-Rong Luo
Alzheimer's disease (AD) is a major public health concern worldwide and the few drugs currently available only treat the symptoms. Hence, there is a strong need to find more effective anti-AD agents. Cynanchum otophyllum is a traditional Chinese medicine for treating epilepsy, and otophylloside B (Ot B), isolated from C. otophyllum, is the essential active component. Having previously identified anti-aging effects of Ot B, we evaluated Ot B for AD prevention in C. elegans models of AD and found that Ot B extended lifespan, increased heat stress-resistance, delayed body paralysis, and increased the chemotaxis response...
February 13, 2017: Natural Products and Bioprospecting
https://www.readbyqxmd.com/read/28194431/polyfunctional-and-ifn-%C3%AE-monofunctional-human-cd4-t-cell-populations-are-molecularly-distinct
#12
Julie G Burel, Simon H Apte, Penny L Groves, James S McCarthy, Denise L Doolan
Pathogen-specific polyfunctional T cell responses have been associated with favorable clinical outcomes, but it is not known whether molecular differences exist between polyfunctional and monofunctional cytokine-producing T cells. Here, we report that polyfunctional CD4(+) T cells induced during Plasmodiumfalciparum (P. falciparum) blood-stage infection in humans have a unique transcriptomic profile compared with IFN-γ monofunctional CD4(+) T cells and, thus, are molecularly distinct. The 14-gene signature revealed in P...
February 9, 2017: JCI Insight
https://www.readbyqxmd.com/read/28194258/complete-genome-sequence-of-pseudomonas-stutzeri-strain-rch2-isolated-from-a-hexavalent-chromium-cr-vi-contaminated-site
#13
Romy Chakraborty, Hannah Woo, Paramvir Dehal, Robert Walker, Marcin Zemla, Manfred Auer, Lynne A Goodwin, Alexey Kazakov, Pavel Novichkov, Adam P Arkin, Terry C Hazen
Hexavalent Chromium [Cr(VI)] is a widespread contaminant found in soil, sediment, and ground water in several DOE sites, including Hanford 100 H area. In order to stimulate microbially mediated reduction of Cr(VI) at this site, a poly-lactate hydrogen release compound was injected into the chromium contaminated aquifer. Targeted enrichment of dominant nitrate-reducing bacteria post injection resulted in the isolation of Pseudomonas stutzeri strain RCH2. P. stutzeri strain RCH2 was isolated using acetate as the electron donor and is a complete denitrifier...
2017: Standards in Genomic Sciences
https://www.readbyqxmd.com/read/28194137/sigma-factor-regulated-cellular-response-in-a-non-solvent-producing-clostridium-beijerinckii-degenerated-strain-a-comparative-transcriptome-analysis
#14
Yan Zhang, Shengyin Jiao, Jia Lv, Renjia Du, Xiaoni Yan, Caixia Wan, Ruijuan Zhang, Bei Han
Clostridium beijerinckii DG-8052, derived from NCIMB 8052, cannot produce solvent or form spores, a phenomenon known as degeneration. To explore the mechanisms of degeneration at the gene level, transcriptomic profiles of the wild-type 8052 and DG-8052 strains were compared. Expression of 5168 genes comprising 98.6% of the genome was assessed. Interestingly, 548 and 702 genes were significantly up-regulated in the acidogenesis and solventogenesis phases of DG-8052, respectively, and mainly responsible for the phosphotransferase system, sugar metabolic pathways, and chemotaxis; meanwhile, 699 and 797 genes were significantly down-regulated, respectively, and mainly responsible for sporulation, oxidoreduction, and solventogenesis...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28193218/a-combined-model-reduction-algorithm-for-controlled-biochemical-systems
#15
Thomas J Snowden, Piet H van der Graaf, Marcus J Tindall
BACKGROUND: Systems Biology continues to produce increasingly large models of complex biochemical reaction networks. In applications requiring, for example, parameter estimation, the use of agent-based modelling approaches, or real-time simulation, this growing model complexity can present a significant hurdle. Often, however, not all portions of a model are of equal interest in a given setting. In such situations methods of model reduction offer one possible approach for addressing the issue of complexity by seeking to eliminate those portions of a pathway that can be shown to have the least effect upon the properties of interest...
February 13, 2017: BMC Systems Biology
https://www.readbyqxmd.com/read/28191768/multimodal-delivery-of-isogenic-mesenchymal-stem-cells-yields-synergistic-protection-from-retinal-degeneration-and-vision-loss
#16
Benjamin Bakondi, Sergey Girman, Bin Lu, Shaomei Wang
We previously demonstrated that subretinal injection (SRI) of isogenic mesenchymal stem cells (MSCs) reduced the severity of retinal degeneration in Royal College of Surgeons rats in a focal manner. In contrast, intravenous MSC infusion (MSC(IV) ) produced panoptic retinal rescue. By combining these treatments, we now show that MSC(IV) supplementation potentiates the MSC(SRI) -mediated rescue of photoreceptors and visual function. Electrophysiological recording from superior colliculi revealed 3.9-fold lower luminance threshold responses (LTRs) and 22% larger functional rescue area from combined treatment compared with MSC(SRI) alone...
February 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28190586/increased-expression-of-glutaminase-in-osteoblasts-promotes-macrophage-recruitment-in-periapical-lesions
#17
Kuo-Liang Hou, Sze-Kwan Lin, Sang-Heng Kok, Han-Wei Wang, Eddie Hsiang-Hua Lai, Chi-Yuan Hong, Hsiang Yang, Juo-Song Wang, Li-Deh Lin, Jenny Zwei-Chieng Chang
INTRODUCTION: Recently, we have shown that tissue hypoxia stimulates the progression of periapical lesions by up-regulating glycolysis-dependent apoptosis of osteoblasts. Other facets of hypoxia-induced metabolic reprogramming in disease pathogenesis require further investigation. In this study, we examined the connection between hypoxia-augmented glutamine catabolism in osteoblasts and the development of periapical lesions. METHODS: Primary human osteoblasts were cultured under hypoxia...
February 9, 2017: Journal of Endodontics
https://www.readbyqxmd.com/read/28188794/surfactant-protein-a-sp-a-and-sp-a-derived-peptide-attenuate-chemotaxis-of-mast-cells-induced-by-human-%C3%AE-defensin-3
#18
Yasuaki Uehara, Motoko Takahashi, Masaki Murata, Atsushi Saito, Rina Takamiya, Yoshihiro Hasegawa, Koji Kuronuma, Hirofumi Chiba, Jiro Hashimoto, Norimasa Sawada, Hiroki Takahashi, Yoshio Kuroki, Shigeru Ariki
Human β-defensin 3 (hBD3) is known to be involved in mast cell activation. However, molecular mechanisms underlying the regulation of hBD3-induced mast cell activation have been poorly understood. We previously reported that SP-A and SP-A-derived peptide 01 (SAP01) regulate the function of hBD3. In this study, we focused on the effects of SP-A and SAP01 on the activation of mast cells induced by hBD3. SAP01 directly bound to hBD3. Mast cell-mediated vascular permeability and edema in hBD3 administered rat ears were decreased when injected with SP-A or SAP01...
February 8, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28186323/the-arp2-3-inhibitory-protein-arpin-is-dispensable-for-chemotaxis
#19
Irène Dang, Joern Linkner, Jun Yan, Daniel Irimia, Jan Faix, Alexis Gautreau
Arpin is an Arp2/3 inhibitory protein, which decreases the protrusion lifetime and hence directional persistence in the migration of diverse cells. Arpin is activated by the small GTPase Rac, which controls cell protrusion, thus closing a negative feedback loop that renders the protrusion intrinsically unstable. Because of these properties, it was proposed that Arpin might play a role in directed migration, where directional persistence has to be fine-tuned. We report here, however, that Arpin depleted tumor cells and Arpin knock-out Dictyostelium amoeba display no obvious defect in chemotaxis...
February 10, 2017: Biology of the Cell
https://www.readbyqxmd.com/read/28186140/activated-niacin-receptor-hca2-inhibits-chemoattractant-mediated-macrophage-migration-via-g%C3%AE-%C3%AE-pkc-erk1-2-pathway-and-heterologous-receptor-desensitization
#20
Ying Shi, Xiangru Lai, Lingyan Ye, Keqiang Chen, Zheng Cao, Wanghua Gong, Lili Jin, Chunyan Wang, Mingyong Liu, Yuan Liao, Ji Ming Wang, Naiming Zhou
The niacin receptor HCA2 is implicated in controlling inflammatory host responses with yet poorly understood mechanistic basis. We previously reported that HCA2 in A431 epithelial cells transduced Gβγ-protein kinase C- and Gβγ-metalloproteinase/EGFR-dependent MAPK/ERK signaling cascades. Here, we investigated the role of HCA2 in macrophage-mediated inflammation and the underlying mechanisms. We found that proinflammatory stimulants LPS, IL-6 and IL-1β up-regulated the expression of HCA2 on macrophages...
February 10, 2017: Scientific Reports
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