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https://www.readbyqxmd.com/read/27921010/structural-insights-into-substrate-recognition-by-clostridium-difficile-sortase
#1
Jui-Chieh Yin, Chun-Hsien Fei, Yen-Chen Lo, Yu-Yuan Hsiao, Jyun-Cyuan Chang, Jay C Nix, Yuan-Yu Chang, Lee-Wei Yang, I-Hsiu Huang, Shuying Wang
Sortases function as cysteine transpeptidases that catalyze the covalent attachment of virulence-associated surface proteins into the cell wall peptidoglycan in Gram-positive bacteria. The substrate proteins targeted by sortase enzymes have a cell wall sorting signal (CWSS) located at the C-terminus. Up to date, it is still not well understood how sortases with structural resemblance among different classes and diverse species of bacteria achieve substrate specificity. In this study, we focus on elucidating the molecular basis for specific recognition of peptide substrate PPKTG by Clostridium difficile sortase B (Cd-SrtB)...
2016: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/27919837/ex-vivo-itreg-differentiation-revisited-convenient-alternatives-to-existing-strategies
#2
Billur Akkaya, Amanda H Holstein, Christopher Isaac, Mitra P Maz, Deborah D Glass, Ethan M Shevach, Munir Akkaya
Ex-vivo differentiation of regulatory T cells (Tregs) from naïve CD4(+) T-cells has been widely used in immunological research. Isolation of a highly pure naïve T cell population is the key factor that determines the efficiency of subsequent Treg differentiation. Currently, this step relies mostly on FACS sorting, which is often costly, time consuming, and inconvenient. Alternatively, magnetic separation of T-cells can be performed; yet, available protocols fail to reach sort level purity and consequently result in low Treg differentiation efficiency...
December 2, 2016: Journal of Immunological Methods
https://www.readbyqxmd.com/read/27919801/folic-acid-capped-pegylated-magnetic-nanoparticles-enter-cancer-cells-mostly-via-clathrin-dependent-endocytosis
#3
Emilie Allard-Vannier, Katel Hervé-Aubert, Karine Kaaki, Thibaut Blondy, Anastasia Shebanova, Konstantin V Shaitan, Anastasia A Ignatova, Marie-Louise Saboungi, Alexey V Feofanov, Igor Chourpa
BACKGROUND: This work is focused on mechanisms of uptake in cancer cells of rationally designed, covalently assembled nanoparticles, made of superparamagnetic iron oxide nanoparticles (SPIONs), fluorophores (doxorubicin or Nile Blue), polyethylene glycol (PEG) and folic acid (FA), referred hereinafter as SFP-FA. METHODS: SFP-FA were characterized by DLS, zetametry and fluorescence spectroscopy. The SFP-FA uptake in cancer cells was monitored using fluorescence-based methods like fluorescence-assisted cell sorting, LSCM with single-photon and two-photon excitation...
December 2, 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27919572/non-fucosylated-cb-cd34-cells-represent-a-good-target-for-enforced-fucosylation-to-improve-engraftment-following-cord-blood-transplantation
#4
Simon N Robinson, Michael W Thomas, Paul J Simmons, Junjun Lu, Hong Yang, Jeannie A Javni, Elizabeth J Shpall, Patrick A Zweidler-Mckay
BACKGROUND AIMS: Despite ethnic diversity and ready availability of cryopreserved, human leukocyte antigen-typed cord blood (CB), delayed engraftment remains a significant hurdle to successful CB transplantation. Suboptimal homing of CB hematopoietic stem and progenitor cells (HSPCs) to the hematopoietic microenvironment (HM) is thought to be responsible and due to low levels of HSPC fucosylation. Fucosylation (decoration with sialyl-Lewis(X)) may improve HSPC homing to HM by increasing the strength of HSPC/E-selectin interactions, where E-selectin is constitutively expressed by HM microvasculature...
December 2, 2016: Cytotherapy
https://www.readbyqxmd.com/read/27919468/somatic-gnaq-mutation-is-enriched-in-brain-endothelial-cells-in%C3%A2-sturge-weber-syndrome
#5
Lan Huang, Javier A Couto, Anna Pinto, Sanda Alexandrescu, Joseph R Madsen, Arin K Greene, Mustafa Sahin, Joyce Bischoff
BACKGROUND: Sturge-Weber syndrome (SWS) is a rare congenital neurocutaneous disorder characterized by facial and extracraniofacial capillary malformations and capillary-venule malformations in the leptomeninges. A somatic mosaic mutation in GNAQ (c.548G>A; p.R183Q) was found in SWS brain and skin capillary malformations. Our laboratory showed endothelial cells in skin capillary malformations are enriched for the GNAQ mutation. The purpose of this study is to determine whether the GNAQ mutation is also enriched in endothelial cells in affected SWS brain...
October 21, 2016: Pediatric Neurology
https://www.readbyqxmd.com/read/27919066/designed-proteins-induce-the-formation-of-nanocage-containing-extracellular-vesicles
#6
Jörg Votteler, Cassandra Ogohara, Sue Yi, Yang Hsia, Una Nattermann, David M Belnap, Neil P King, Wesley I Sundquist
Complex biological processes are often performed by self-organizing nanostructures comprising multiple classes of macromolecules, such as ribosomes (proteins and RNA) or enveloped viruses (proteins, nucleic acids and lipids). Approaches have been developed for designing self-assembling structures consisting of either nucleic acids or proteins, but strategies for engineering hybrid biological materials are only beginning to emerge. Here we describe the design of self-assembling protein nanocages that direct their own release from human cells inside small vesicles in a manner that resembles some viruses...
November 30, 2016: Nature
https://www.readbyqxmd.com/read/27918526/mycobacterium-tuberculosis-esxh-inhibits-escrt-dependent-cd4-t-cell-activation
#7
Cynthia Portal-Celhay, JoAnn M Tufariello, Smita Srivastava, Aleena Zahra, Thais Klevorn, Patricia S Grace, Alka Mehra, Heidi S Park, Joel D Ernst, William R Jacobs, Jennifer A Philips
Mycobacterium tuberculosis (Mtb) establishes a persistent infection, despite inducing antigen-specific T-cell responses. Although T cells arrive at the site of infection, they do not provide sterilizing immunity. The molecular basis of how Mtb impairs T-cell function is not clear. Mtb has been reported to block major histocompatibility complex class II (MHC-II) antigen presentation; however, no bacterial effector or host-cell target mediating this effect has been identified. We recently found that Mtb EsxH, which is secreted by the Esx-3 type VII secretion system, directly inhibits the endosomal sorting complex required for transport (ESCRT) machinery...
December 5, 2016: Nature Microbiology
https://www.readbyqxmd.com/read/27917441/size-selective-harvesting-of-extracellular-vesicles-for-strategic-analyses-towards-tumor-diagnoses
#8
Chihiro Manri, Takahide Yokoi, Hirokazu Nishida
Extracellular vesicles (EV), typified by exosomes or microvesicles, are expected to be effective diagnostic markers for cancers. The sizes of the vesicles range from 20 to 1000 nm, but the size-dependent variations of the contents of EVs are still poorly understood. We succeeded in the size-selective harvesting of the vesicles by utilizing the molecular weight-dependent characteristics of a variety of polyethylene glycols (PEG) as precipitating reagents and analyzed the antigens displayed on the surfaces of the vesicles and the miRNAs included in the vesicles from each size group...
December 5, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27917337/identification-characterization-and-phylogenetic-analysis-of-antifungal-trichoderma-from-tomato-rhizosphere
#9
Shalini Rai, Prem Lal Kashyap, Sudheer Kumar, Alok Kumar Srivastava, Pramod W Ramteke
The use of Trichoderma isolates with efficient antagonistic activity represents a potentially effective and alternative disease management strategy to replace health hazardous chemical control. In this context, twenty isolates were obtained from tomato rhizosphere and evaluated by their antagonistic activity against four fungal pathogens (Fusarium oxysporum f. sp. lycopersici, Alternaria alternata, Colletotrichum gloeosporoides and Rhizoctonia solani). The production of extracellular cell wall degrading enzymes of tested isolates was also measured...
2016: SpringerPlus
https://www.readbyqxmd.com/read/27917252/separation-of-superparamagnetic-particles-through-ratcheted-brownian-motion-and-periodically-switching-magnetic-fields
#10
Fan Liu, Li Jiang, Huei Ming Tan, Ashutosh Yadav, Preetika Biswas, Johan R C van der Maarel, Christian A Nijhuis, Jeroen A van Kan
Brownian ratchet based particle separation systems for application in lab on chip devices have drawn interest and are subject to ongoing theoretical and experimental investigations. We demonstrate a compact microfluidic particle separation chip, which implements an extended on-off Brownian ratchet scheme that actively separates and sorts particles using periodically switching magnetic fields, asymmetric sawtooth channel sidewalls, and Brownian motion. The microfluidic chip was made with Polydimethylsiloxane (PDMS) soft lithography of SU-8 molds, which in turn was fabricated using Proton Beam Writing...
November 2016: Biomicrofluidics
https://www.readbyqxmd.com/read/27917174/cd16-monocyte-subset-was-enriched-and-functionally-exacerbated-in-driving-t-cell-activation-and-b-cell-response-in-systemic-lupus-erythematosus
#11
Huaqun Zhu, Fanlei Hu, Xiaolin Sun, Xiaoying Zhang, Lei Zhu, Xu Liu, Xue Li, Liling Xu, Lianjie Shi, Yuzhou Gan, Yin Su
BACKGROUND: The roles that CD16(+) monocyte subset plays in T-cell activation and B-cell response have not been well studied in systemic lupus erythematosus (SLE). OBJECTIVE: The present study aimed to investigate the distribution of CD16(+) monocyte subsets in SLE and explore their possible roles in T-cell activation and B-cell differentiation. METHODS: The frequencies of monocyte subsets in the peripheral blood of healthy controls (HCs) and patients with SLE were determined by flow cytometry...
2016: Frontiers in Immunology
https://www.readbyqxmd.com/read/27917009/induced-expression-of-cancer-stem-cell-markers-aldh1a3-and-sox-2-in-hierarchical-reconstitution-of-apoptosis-resistant-human-breast-cancer-cells
#12
Karin Kashii-Magaribuchi, Rie Takeuchi, Yuko Haisa, Akemi Sakamoto, Aimi Itoh, Yuki Izawa, Miyuki Isa, Mayu Fukuzawa, Motonobu Murakami, Rei Takahashi
We established an experimental system that can induce p53-dependent apoptosis by doxycycline treatment to analyze characteristics of the apoptosis-resistant cancer cell subpopulation in the human breast cancer cell line HCC1937. Expression patterns of the stem cell markers, ALDH1A3 and Sox-2, the luminal differentiation marker, GATA3 and the proliferation index marker, Ki-67 were analyzed using immunostaining and fluorescence-activated cell sorting (FACS). After doxycycline treatment, the number of viable cells was gradually decreased over seven days in a time-dependent manner due to p53-induced apoptosis; however, the number of smaller-sized ALDH1A3(+) cells assessed by immunostaining increased sharply after 1 day of doxycycline treatment, suggesting their apoptosis-resistant nature...
November 1, 2016: Acta Histochemica et Cytochemica
https://www.readbyqxmd.com/read/27916908/epidermal-growth-factor-receptor-status-in-circulating-tumor-cells-as-a-predictive-biomarker-of-sensitivity-in-castration-resistant-prostate-cancer-patients-treated-with-docetaxel-chemotherapy
#13
Takatsugu Okegawa, Naoshi Itaya, Hidehiko Hara, Mitsuhiro Tambo, Kikuo Nutahara
OBJECTIVE: We examined whether epidermal growth factor receptor (EGFR) expression in circulating tumor cells (CTCs) can be used to predict survival in a population of bone-metastatic castration-resistant prostate cancer (mCRPC) patients treated with docetaxel chemotherapy. METHODS: All patients with mCRPC who had experienced treatment failure with androgen-deprivation therapy and had received docetaxel chemotherapy were eligible. CTCs and EGFR expression in CTCs were enumerated with the CellSearch System in whole blood...
November 30, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27916087/-the-regulatory-role-of-wnt-signaling-pathway-in-the-proliferation-and-activation-of-memory-t-cells
#14
Lei Luo, Guangheng Luo, Qian Fang, Shan Huang, Hui Chen
Objective To study the regulatory effect of Wnt signaling pathway on the activation and proliferation of memory T cells (TMs). Methods We established the mouse model of skin allograft transplantation (BALB/c-C57BL/6), and then we sorted and cultured immature dendritic cells (imDCs), mature dendritic cells (mDCs) and TMs from the C57BL/6 mice by immunomagnetic bead cell sorting. The imDCs or mDCs loading antigens from BALB/c mice were co-cultured with TMs in a 96-well plate or Transwell(TM) plate (MLR). The (3)H-TdR was used to detect the proliferation of TMs...
December 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/27915349/roles-of-1-25-oh-2d3-and-vitamin-d-receptor-in-the-pathogenesis-of-rheumatoid-arthritis-and-systemic-lupus-erythematosus-by-regulating-the-activation-of-cd4-t-cells-and-the-pkc%C3%AE-erk-signaling-pathway
#15
Xiao-Jie He, Yan Ding, Wei Xiang, Xi-Qiang Dang
BACKGROUND/AIMS: The study aims to elucidate the roles of 1,25(OH)2D3 and vitamin D receptor (VDR) in the pathogenesis of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) by regulating the activation of CD4+ T cells and the PKCδ/ERK signaling pathway. METHODS: From January 2013 to December 2015, a total of 130 SLE patients, 137 RA patients and 130 healthy controls were selected in this study. Serum levels of 1,25(OH)2D3 and VDR mRNA expression were detected by ELISA and real-time fluorescence quantitative PCR (RT-qPCR)...
December 5, 2016: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/27911709/shuttling-and-sorting-lipid-modified-cargo-into-the-cilia
#16
REVIEW
Louise A Stephen, Shehab Ismail
Primary cilia are hair-like microtubule-based organelles that can be found on almost all human cell types. Although the cilium is not separated from the cell by membranes, their content is different from that of the cell body and their membrane composition is distinct from that of the plasma membrane. Here, we will introduce a molecular machinery that shuttles and sorts lipid-modified proteins to the cilium, thus contributing in maintaining its distinct composition. The mechanism involves the binding of the GDI-like solubilising factors, uncoordinated (UNC)119a, UNC119b and PDE6D, to the lipid-modified ciliary cargo and the specific release of the cargo in the cilia by the ciliary small G-protein Arl3 in a GTP-dependent manner...
October 15, 2016: Biochemical Society Transactions
https://www.readbyqxmd.com/read/27911272/foxo4-expression-is-related-to-stem-cell-like-properties-and-resistance-to-treatment-in-diffuse-large-b-cell-lymphoma
#17
Kyung Ju Ryu, Chaehwa Park, Mineui Hong, Young Hyeh Ko, Won Seog Kim, Seok Jin Kim
Cancer stem cells are proposed to be responsible for resistance to chemotherapeutic agents, including doxorubicin. As phenylbutyrate enhances cancer stem cell properties, we analyzed surviving lymphoma cells after treatment with doxorubicin and phenylbutyrate. Human B-cell lymphoma cell lines, including Toledo, BJAB, Daudi, and Raji were incubated with IC90 concentrations of doxorubicin (300 nM) or phenylbutyrate (8 mM). After 48 h, live cells were sorted and analyzed for their resistance to treatment by examining gene expression profiles using cDNA microarray and biological characteristics...
November 29, 2016: Oncotarget
https://www.readbyqxmd.com/read/27910898/hydrogen-overproducing-nitrogenases-obtained-by-random-mutagenesis-and-high-throughput-screening
#18
Emma Barahona, Emilio Jiménez-Vicente, Luis M Rubio
When produced biologically, especially by photosynthetic organisms, hydrogen gas (H2) is arguably the cleanest fuel available. An important limitation to the discovery or synthesis of better H2-producing enzymes is the absence of methods for the high-throughput screening of H2 production in biological systems. Here, we re-engineered the natural H2 sensing system of Rhodobacter capsulatus to direct the emission of LacZ-dependent fluorescence in response to nitrogenase-produced H2. A lacZ gene was placed under the control of the hupA H2-inducible promoter in a strain lacking the uptake hydrogenase and the nifH nitrogenase gene...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27910751/recent-advances-in-pharmacotherapeutic-paradigm-of-mild-to-recalcitrant-atopic-dermatitis
#19
Zahid Hussain, Shariza Sahudin, Hnin Ei Thu, Ahmad Nazrun Shuid, Syed Nasir Abbas Bukhari, Endang Kumolosasi
Atopic dermatitis (AD) is a common, chronic skin inflammatory disorder characterized by perivascular infiltration of immunoglobulin E (IgE), T lymphocytes, and mast cells. The key factors responsible for the pathophysiology of this disease are immunological disorders and defects in epidermal barrier properties. Pruritus, intense itching, psychological stress, deprived physical and mental performance, and sleep disturbance are the hallmark features of this dermatological disorder. Preventive interventions such as educational programs, avoidance of allergens, and exclusive care toward the skin could play a partial role in suppressing the symptoms...
2016: Critical Reviews in Therapeutic Drug Carrier Systems
https://www.readbyqxmd.com/read/27910289/visible-laser-induced-in-situ-cell-detachment-from-gold-nanoparticle-embedded-collagen-gel
#20
Chie Kojima, Yusuke Nakajima, Naoya Oeda, Takeshi Kawano, Yusuke Taki
Cell sorting is important for cell biology and regenerative medicine. A visible light-responsive cell scaffold is produced using gold nanoparticles and collagen gel. Various kinds of cells are cultured on the visible light-responsive cell scaffold, and the target cells are selectively detached by photoirradiation without any cytotoxicity. This is a new image-guided cell sorting system.
December 2, 2016: Macromolecular Bioscience
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