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https://www.readbyqxmd.com/read/29786968/p62-mediated-autophagy-affects-nutrition-dependent-insulin-receptor-substrate-1-dynamics-in-3t3-l1-preadipocytes
#1
Hirobumi Igawa, Akihiro Kikuchi, Hirofumi Misu, Kiyo-Aki Ishii, Shuichi Kaneko, Toshinari Takamura
AIMS/INTRODUCTION: Previous studies have shown that the organism's nutritional status changes the protein levels of insulin receptor substrate 1 (IRS-1) in a tissue-specific manner. Although the mechanisms underlying the regulation of IRS-1 in the nutrient-rich conditions associated with diabetes and insulin resistance have been well studied, those under nutrient-poor conditions remain unknown. The aim of this study was to investigate how IRS-1 protein levels change depending on the nutritional status of 3T3-L1 preadipocytes...
May 22, 2018: Journal of Diabetes Investigation
https://www.readbyqxmd.com/read/29786785/methods-to-study-angiogenesis-in-a-mouse-model-of-prostate-cancer
#2
Ana-Rita Pedrosa, Alexandre Trindade, António Duarte
Angiogenesis is one important hallmark of cancer progression which explains the relevance of developing methods to efficiently analyze the neo-angiogenic process. In this report we make use of the transgenic adenocarcinoma of the murine prostate (TRAMP) model, considered a good model for studying clinical prostate cancer progression, to describe in detail the methods used to study angiogenesis in this type of solid tumor development. In this report we provide step-by-step procedures on the basis of previous work in our laboratory for: the mouse urogenital sinus (UGS) collection; microdissection of the prostate; preparation of the prostatic samples for immunofluorescence (to analyze vascular density, morphology, maturation, functionality, hypoxia, and others); preparation of prostatic samples to histopathological analysis and/or immunohistochemistry; and endothelial and vascular mural cell sorting and isolation by fluorescent associated cell sorting (FACS) to further analysis (mRNA, protein, or other) or to maintain in culture...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29786639/chemogenetic-enhancement-of-axon-regeneration-following-peripheral-nerve-injury-in-the-slick-a-mouse
#3
Poonam B Jaiswal, Olivia C Mistretta, Patricia J Ward, Arthur W English
The effects of chemogenetics on axon regeneration following peripheral nerve transection and repair were studied in mice expressing a Cre-dependent excitatory designer receptor exclusively activated by designer drugs (DREADD) and Cre-recombinase/yellow fluorescent protein (YFP) in a subset of motor and sensory neurons and cortical motoneurons (SLICK-A). Sciatic nerves were cut and repaired and mice were treated either once, at the time of injury, or five days per week for two weeks with clozapine N-oxide (CNO) (1 mg/kg, i...
May 22, 2018: Brain Sciences
https://www.readbyqxmd.com/read/29786602/microfluidic-system-for-modelling-3d-tumour-invasion-into-surrounding-stroma-and-drug-screening
#4
Zhichang Du, Shengli Mi, Xiaoman Yi, Yuanyuan Xu, Wei Sun
Tumour invasion into the surrounding stroma is a critical step in metastasis, and it is necessary to clarify the role of microenvironmental factors in tumour invasion. We present a microfluidic system that simulated and controlled multi-factors of the tumour microenvironment for three-dimensional (3D) assessment of tumour invasion into the stroma. The simultaneous, precise and continuous arrangement of two 3D matrices was visualised to observe the migration of cancer cell populations or single cells by transfecting cells with a fluorescent protein...
May 22, 2018: Biofabrication
https://www.readbyqxmd.com/read/29785968/force-activatable-coating-enables-high-resolution-cellular-force-imaging-directly-on-regular-cell-culture-surfaces
#5
Anwesha Sarkar, Yuanchang Zhao, Yongliang Wang, Xuefeng Wang
Integrin-transmitted cellular forces are crucial mechanical signals regulating a vast range of cell functions. Although various methods have been developed to visualize and quantify cellular forces at the cell-matrix interface, a method with high performance and low technical barrier is still in demand. Here we developed force-activatable coating (FAC) which can be simply coated on regular cell culture apparatus's surfaces by physical adsorption, and turn these surfaces to force reporting platforms that enable cellular force mapping directly by fluorescence imaging...
May 22, 2018: Physical Biology
https://www.readbyqxmd.com/read/29785699/spectroscopic-analysis-of-the-cu-2-induced-fluorescence-quenching-of-fluorescent-proteins-amcyan-and-morange2
#6
Ji-Eun Bae, In Jung Kim, Ki Hyun Nam
Fluorescent proteins show fluorescence quenching by specific metal ions, which can be applied towards metal biosensing applications. In order to develop metal-biosensor, we performed spectroscopic analysis of the fluorescence quenching of fluorescent protein AmCyan and mOrange2 by various metal ions. The fluorescence intensity of AmCyan was reduced to 48.54% by Co2+ and 67.77% by Zn2+ ; Cu2+ reduced the fluorescence emission of AmCyan to 19.30% of its maximum. The fluorescence intensity of mOrange2 was quenched by only Cu2+ , to 11...
May 21, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29785600/performance-and-microbial-community-of-cic-anaerobic-reactor-treating-food-waste-under-different-grease-contents-and-inner-circulation-ratio
#7
Chengyuan Su, Lijian Zhao, Liming Liao, Jingjing Qin, Yuxiang Lu, Menglin Chen, Mei Huang, Zhi Huang
High concentrations of grease easily inhibit anaerobic digestion. The stability of the process and microbial responses in the controlling internal circulation (CIC) reactor used for treating food waste were investigated under different grease contents and inner circulation ratios. Results showed that at the grease content of 1 g/L, the removal rates of 94% and 86-93% were achieved for chemical oxygen demand (COD) and NH3 -N, respectively. In contrast, when the grease content increased to 7 g/L, removal rates for COD and NH3 -N significantly decreased to 42...
May 21, 2018: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/29785487/rapid-monitoring-of-the-target-protein-expression-with-a-fluorescent-signal-based-on-a-dicistronic-construct-in-escherichia-coli
#8
Chang-Ye Hui, Yan Guo, Wen Zhang, Xian-Qing Huang
Real-time quantification of recombinant proteins is important in studies on fermentation engineering, cell engineering, etc. Measurement of the expression level of heterologous proteins in bacterial fermentation broth has traditionally relied on time-consuming and labor-intensive procedures, such as polyacrylamide gel electrophoresis, immunoblot analysis, and biological activity assays. We describe a simple, fast, and high sensitive assay for detecting heterologous proteins production in bacteria either at the overall level (fluorescence spectrophotometry) or at the individual level (fluorescence microscopic image) in this study...
May 21, 2018: AMB Express
https://www.readbyqxmd.com/read/29785357/prognostic-significance-of-cyclins-a2-b1-d1-and-e1-and-ccnd1-numerical-aberrations-in-oral-squamous-cell-carcinomas
#9
Luís Silva Monteiro, Márcio Diniz-Freitas, Saman Warnakulasuriya, Tomás Garcia-Caballero, Jerónimo Forteza-Vila, Máximo Fraga
We analysed the expression of cyclins A2, B1, D1, and E1 by immunohistochemistry and numerical aberrations in CCND1 gene by fluorescence in situ hybridization technique in 67 primary oral squamous cell carcinomas (OSCC). Cyclin A2 expression was observed in 54 (83.1%) tumours, cyclin D1 in 58 (89.2%), cyclin B1 in 39 (60%), and cyclin E in 21 (32.8%). CCND1 region analysis revealed 26 (43.3%) tumours with the presence of numerical aberrations which were correlated with cyclin D1 high expression (Rho = 0...
2018: Analytical Cellular Pathology (Amsterdam)
https://www.readbyqxmd.com/read/29785139/blockage-of-store-operated-ca-2-entry-antagonizes-epstein-barr-virus-promoted-angiogenesis-by-inhibiting-ca-2-signaling-regulated-vegf-production-in-nasopharyngeal-carcinoma
#10
Jiaxiang Ye, Junqi Huang, Qian He, Weilin Zhao, Xiaoying Zhou, Zhe Zhang, Yongqiang Li, Jiazhang Wei, Jinyan Zhang
Background: Epstein-Barr virus (EBV) actively contributes to the pathological process of nasopharyngeal carcinoma (NPC) by enabling NPC cells to acquire various capacities required for their malignant biological actions. Our earlier works demonstrated that EBV-encoded latent membrane protein 1 (LMP1) enhanced vascular endothelial growth factor (VEGF)-mediated angiogenesis by boosting store-operated Ca2+ entry (SOCE) upon extracellular epidermal growth factor (EGF) stimulation. However, the antagonistic effects of SOCE blockage on EBV-promoted angiogenesis must be appropriately evaluated in vivo, and the global effect of EBV infection on the EGF-elicited cytosolic Ca2+ signaling, which regulates VEGF-mediated angiogenesis remains to be further clarified...
2018: Cancer Management and Research
https://www.readbyqxmd.com/read/29784884/the-effector-gene-xopae-of-xanthomonas-euvesicatoria-85-10-is-part-of-an-operon-and-encodes-an-e3-ubiquitin-ligase
#11
Georgy Popov, Bharat Bhusan Majhi, Guido Sessa
The type III effector XopAE from the Xanthomonas euvesicatoria strain 85-10 ( Xe 85-10) was previously shown to inhibit plant immunity and enhance pathogen-induced disease symptoms. Evolutionary analysis of 60 xopAE alleles ( AEal ) revealed that the xopAE locus is conserved in multiple Xanthomonas species. The majority of xopAE alleles (55 out of 60) encodes a single ORF ( xopAE ), while in 5 alleles, including AEal 37 of the Xe 85-10 strain, a frame-shift splits the locus into two ORFs ( hpaF and a truncated xopAE )...
May 21, 2018: Journal of Bacteriology
https://www.readbyqxmd.com/read/29784874/select-%C3%AE-arrestins-control-cell-surface-abundance-of-the-mammalian-kir2-1-potassium-channel-in-a-yeast-model
#12
Natalie A Hager, Collin J Krasowski, Timothy D Mackie, Alexander R Kolb, Patrick G Needham, Andrew A Augustine, Alison Dempsey, Christopher Szent-Gyorgyi, Marcel P Bruchez, Daniel J Bain, Adam V Kwiatkowski, Allyson F O'Donnell, Jeffrey L Brodsky
Protein composition at the plasma membrane is tightly regulated, with rapid protein internalization and selective targeting to the cell surface occurring in response to environmental changes. For example, ion channels are dynamically relocalized to or from the plasma membrane in response to physiological alterations, allowing cells and organisms to maintain osmotic and salt homeostasis. To identify additional factors that regulate the selective trafficking of a specific ion channel, we used a yeast model for a mammalian potassium channel, the K+ inwardly rectifying channel Kir2...
May 21, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29784858/production-of-staphylococcal-complement-inhibitor-scin-and-other-immune-modulators-during-the-early-stages-of-staphylococcus-aureus-biofilm-formation-in-a-mammalian-cell-culture-medium
#13
Andi R Sultan, Jasper W Swierstra, Nicole A Lemmens-den Toom, Susan V Snijders, Silvie Hansenová Maňásková, Annelies Verbon, Willem J B van Wamel
Immune modulators are known to be produced by matured biofilms and during different stages of planktonic growth of Staphylococcus aureus. Little is known about immune modulator production during the early stages of biofilm formation, thus raising the question: how does S. aureus protect itself from the innate immune responses at these stages? Therefore, we determined the production of the following immune modulators: chemotaxis inhibitory protein of staphylococci (CHIPS), staphylococcal complement inhibitor (SCIN), formyl peptide receptor-like 1 inhibitor, gamma-hemolysin component B, leukocidin D, E, and S, staphylococcal superantigen-like protein 1, 3, 5 and 9, and staphylococcal enterotoxin A...
May 21, 2018: Infection and Immunity
https://www.readbyqxmd.com/read/29784853/carbapenem-non-susceptible-haemophilus-influenzae-with-penicillin-binding-protein-3-containing-amino-acid-insertion
#14
Kazuki Kitaoka, Kouji Kimura, Hiromitsu Kitanaka, Hirotsugu Banno, Wanchun Jin, Jun-Ichi Wachino, Yoshichika Arakawa
Prevalence of β-lactamase-negative ampicillin-resistant Haemophilus influenzae (BLNAR) has become a clinical concern. In BLNAR, amino acid substitutions in penicillin-binding protein 3 (PBP3) are relevant to the β-lactam resistance. Carbapenem-non-susceptible H. influenzae isolates have been rarely reported. Through antimicrobial susceptibility testing and nucleotide sequence analysis of ftsI , encoding PBP3, and utilizing a collection of H. influenzae clinical isolates in our laboratory, we obtained a carbapenem-non-susceptible clinical isolate (NUBL1772) that possessed an altered PBP3 containing V525_N526insM...
May 21, 2018: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/29784393/interactions-of-pyrene-and-or-1-hydroxypyrene-with-bovine-serum-albumin-based-on-eem-parafac-combined-with-molecular-docking
#15
Jing Zhang, Linfeng Chen, Dan Liu, Yaxian Zhu, Yong Zhang
The interactions of pyrene (Pyr) and/or 1-hydroxypyrene (1-OHPyr) with bovine serum albumin (BSA) in binary and ternary systems were investigated using the excitation-emission matrix (EEM)-parallel factor analysis (PARAFAC) method combined with fluorescence quenching analysis and the molecular docking method. The results showed that the PARAFAC approach could be used to decompose the EEM spectra of Pyr, 1-OHPyr, and BSA in the binary and ternary systems. The binding constants of Pyr and 1-OHPyr with BSA increased from 1...
August 15, 2018: Talanta
https://www.readbyqxmd.com/read/29783874/influence-of-bovine-serum-albumin-pre-incubation-on-toxicity-and-er-stress-apoptosis-gene-expression-in-thp-1-macrophages-exposed-to-zno-nanoparticles
#16
Hongying Liang, Tong He, Jimin Long, Liangliang Liu, Guochao Liao, Yanhuai Ding, Yi Cao
When entering a biological environment, proteins could be adsorbed onto nanoparticles (NPs), which will consequently influence the toxicity of NPs. This study used bovine serum albumin (BSA) as the model for serum protein and investigated its interactions with three different types of ZnO NPs, coded as XFI06 (pristine NPs of 20 nm), NM110 (pristine NPs of 100 nm) and NM111 (hydrophobic NPs of 130 nm). Atomic force microscope indicated the adsorption of BSA to ZnO NPs, leading to the increase of NP diameters...
May 21, 2018: Toxicology Mechanisms and Methods
https://www.readbyqxmd.com/read/29783753/broad-spectrum-antimicrobial-activity-and-low-cytotoxicity-against-human-cells-of-a-peptide-derived-from-bovine-%C3%AE-s1-casein
#17
Juncai Hou, Zhijing Liu, Songsong Cao, Haimei Wang, Chenggang Jiang, Muhammad Altaf Hussain, Shiyue Pang
The primary objective of this study was to improve our understanding of the antimicrobial mechanism of protein-derived peptides and to provide evidence for protein-derived peptides as food bio-preservatives by examining the antimicrobial activities, low cytotoxicity, stabilities, and mechanism of Cp1 (LRLKKYKVPQL). In this study, the protein-derived peptide Cp1 was synthesized from bovine αS1 -casein, and its potential use as a food biopreservative was indicated by the higher cell selectivity shown by 11-residue peptide towards bacterial cells than human RBCs...
May 19, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29783744/interplay-between-er-ca-2-binding-proteins-stim1-and-stim2-is-required-for-store-operated-ca-2-entry
#18
Heather A Nelson, Colin A Leech, Richard F Kopp, Michael W Roe
Store-operated calcium entry (SOCE), a fundamentally important homeostatic and Ca2+ signaling pathway in many types of cells, is activated by the direct interaction of stromal interaction molecule 1 (STIM1), an endoplasmic reticulum (ER) Ca2+ -binding protein, with Ca2+ -selective Orai1 channels localized in the plasma membrane. While much is known about the regulation of SOCE by STIM1, the role of stromal interaction molecule 2 (STIM2) in SOCE remains incompletely understood. Here, using clustered regularly interspaced short palindromic repeats -CRISPR associated protein 9 (CRISPR-Cas9) genomic editing and molecular imaging, we investigated the function of STIM2 in NIH 3T3 fibroblast and αT3 cell SOCE...
May 19, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29783743/cell-propagation-of-cholera-toxin-cta-adp-ribosylating-factor-by-exosome-mediated-transfer
#19
Cristiana Zanetti, Angelo Gallina, Alessia Fabbri, Sofia Parisi, Angela Palermo, Katia Fecchi, Zaira Boussadia, Maria Carollo, Mario Falchi, Luca Pasquini, Maria Luisa Fiani, Massimo Sargiacomo
In this study, we report how the cholera toxin (CT) A subunit (CTA), the enzyme moiety responsible for signaling alteration in host cells, enters the exosomal pathway, secretes extracellularly, transmits itself to a cell population. The first evidence for long-term transmission of CT's toxic effect via extracellular vesicles was obtained in Chinese hamster ovary (CHO) cells. To follow the CT intracellular route towards exosome secretion, we used a novel strategy for generating metabolically-labeled fluorescent exosomes that can be counted by flow cytometry assay (FACS) and characterized...
May 19, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29783198/toxicity-assessment-of-metal-oxide-nano-pollutants-on-tomato-solanum-lycopersicon-a-study-on-growth-dynamics-and-plant-cell-death
#20
Bilal Ahmed, Mohammad Saghir Khan, Javed Musarrat
The present study for the first time demonstrated the interactions of metal oxide (MO) nano-pollutants (CuO and Al2 O3 -NPs) with tissues and cellular DNA of tomato plants grown in soil sand: silt: clay (667:190:143) and Hoagland-hydroponic system and assessed the hazardous effects of NPs on cell physiology and biochemistry. Results of SEM equipped with EDX revealed attachment of variably shaped CuO-NPs (18 nm) and Al2 O3 -NPs (21 nm) on roots, and internalization followed by translocation in plants by ICP-MS and TEM...
May 18, 2018: Environmental Pollution
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