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https://www.readbyqxmd.com/read/27914090/measuring-antibody-orientation-at-the-bacterial-surface
#1
Oonagh Shannon, Pontus Nordenfelt
Many bacteria have the ability to interact with antibodies as a means to circumvent the immune response. This includes binding to the Fc portion of antibodies, effectively reversing the antibody orientation and thus decreasing the Fc-mediated immune signaling. Since antibody orientation at the bacterial surface has been shown to be important in human disease, it is valuable to be able to assess how antibodies are interacting with bacterial pathogens. Here, we describe a method to measure the proportion of human IgG that are bound via their Fc or Fabs to a bacterial surface...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914082/detection-of-intracellular-proteins-by-high-resolution-immunofluorescence-microscopy-in-streptococcus-pyogenes
#2
Assaf Raz
Immunofluorescence microscopy is an invaluable tool for the study of biological processes at the cellular level. While the localization of surface-exposed antigens can easily be determined using fluorescent antibodies, localization of intracellular antigens requires permeabilization of the bacterial cell wall and membrane. Here, we describe an immunofluorescence protocol tailored specifically for Streptococcus pyogenes, applying the phage lysin PlyC for cell wall permeabilization. This protocol allows a high level of morphological preservation, suitable for high-resolution microscopy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914080/immobilization-techniques-of-bacteria-for-live-super-resolution-imaging-using-structured-illumination-microscopy
#3
Amy L Bottomley, Lynne Turnbull, Cynthia B Whitchurch, Elizabeth J Harry
Advancements in optical microscopy technology have allowed huge progression in the ability to understand protein structure and dynamics in live bacterial cells using fluorescence microscopy. Paramount to high-quality microscopy is good sample preparation to avoid bacterial cell movement that can result in motion blur during image acquisition. Here, we describe two techniques of sample preparation that reduce unwanted cell movement and are suitable for application to a number of bacterial species and imaging methods...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914077/making-fluorescent-streptococci-and-enterococci-for-live-imaging
#4
Sarah Shabayek, Barbara Spellerberg
Since the discovery of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria, outstanding fluorescent labeling tools with numerous applications in vastly different areas of life sciences have been developed. To optimize GFP for diverse life science applications, a large variety of GFP derivatives with different environmental characteristics have been generated by mutagenesis. The enhanced green fluorescent protein (EGFP) is a well-known GFP derivative with highly increased fluorescence intensity compared to the GFP wild-type molecule...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27914067/5q-syndrome-like-features-as-the-first-manifestation-of-myelodysplastic-syndrome-in-a-patient-with-an-unbalanced-whole-arm-translocation-der-5-19-p10-q10
#5
Hiroshi Ureshino, Haruna Kizuka, Kana Kusaba, Haruhiko Sano, Atsujiro Nishioka, Takero Shindo, Yasushi Kubota, Toshihiko Ando, Kensuke Kojima, Shinya Kimura
Derivative (5;19)(p10;q10) [der(5;19)(p10;q10)] is a rare chromosomal abnormality in myelodysplastic syndrome (MDS), and is genetically similar to deletion 5q [del(5q)]. However, MDS with der(5;19)(p10;q10) and 5q- syndrome are generally characterized as distinct subtypes. Here, we report a case of a patient with 5q- syndrome-like features as the first manifestation of MDS with der(5; 19)(p10;q10). A 59-year-old woman was admitted to our hospital for anemia without leukopenia and thrombocytopenia. She had received chemotherapy comprising carboplatin and docetaxel for endometrial cancer eight years before...
December 2, 2016: International Journal of Hematology
https://www.readbyqxmd.com/read/27913947/preclinical-evaluation-of-cathepsin-based-fluorescent-imaging-system-for-cytoreductive-surgery
#6
Carlos H F Chan, Lukas F Liesenfeld, Isabel Ferreiro-Neira, James C Cusack
BACKGROUND: Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS-HIPEC) is a treatment option for peritoneal surface malignancies. The ability to detect microscopic foci of peritoneal metastasis intraoperatively may ensure the completeness of cytoreduction. In this study, we evaluated the suitability of a hand-held cathepsin-based fluorescent imaging system for intraoperative detection of appendiceal and colorectal peritoneal metastasis. METHODS: Peritoneal tumors and normal peritoneal tissues were collected from patients with appendiceal and colorectal peritoneal metastasis...
December 2, 2016: Annals of Surgical Oncology
https://www.readbyqxmd.com/read/27913886/influence-of-vessel-stenosis-on-indocyanine-green-fluorescence-intensity-assessed-by-near-infrared-fluorescence-angiography
#7
Masaki Yamamoto, Hideaki Nishimori, Takashi Fukutomi, Takemi Handa, Kazuki Kihara, Miwa Tashiro, Takayuki Sato, Kazumasa Orihashi
PURPOSE: Although useful for visualizing blood flow during revascularization surgery, the permeability of near-infrared fluorescence (NIR) angiography using indocyanine green (ICG) does not allow for vessel stenosis visualization. We hypothesized that changes in ICG fluorescence intensity reflect vessel stenosis, and evaluated the influence of stenosis on blood flow by ex vivo experimentation. METHODS: The vessel stenosis model comprised a silicon tube, a graft occluder, and artificial blood...
December 2, 2016: Surgery Today
https://www.readbyqxmd.com/read/27913867/comparison-of-different-concepts-for-interpretation-of-chromosomal-aberrations-in-urothelial-cells-detected-by-fluorescence-in-situ-hybridization
#8
Johannes Mischinger, Lutz Philipp Guttenberg, Jörg Hennenlotter, Georgios Gakis, Stefan Aufderklamm, Steffen Rausch, Eva Neumann, Jens Bedke, Stefan Kruck, Christian Schwentner, Arnulf Stenzl, Tilman Todenhöfer
PURPOSE: Urine fluorescence in situ hybridization (FISH) has become a broadly used marker for noninvasive detection of bladder cancer (BC). However, it has been discussed whether the interpretation algorithm proposed by the manufacturer could be improved. Aim of the present study was to compare alternative evaluation strategies of FISH for detection of BC. METHODS: We included 1048 patients suspicious for BC, who underwent urine FISH examination before cystoscopy (diagnostic cohort)...
December 2, 2016: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/27913845/mitogen-activated-protein-kinases-are-involved-in-hepatocanalicular-dysfunction-and-cholestasis-induced-by-oxidative-stress
#9
Flavia D Toledo, Cecilia L Basiglio, Ismael R Barosso, Andrea C Boaglio, Andrés E Zucchetti, Enrique J Sánchez Pozzi, Marcelo G Roma
In previous studies, we showed that the pro-oxidant model agent tert-butyl hydroperoxide (tBuOOH) induces alterations in hepatocanalicular secretory function by activating Ca(2+)-dependent protein kinase C isoforms (cPKC), via F-actin disorganization followed by endocytic internalization of canalicular transporters relevant to bile formation (Mrp2, Bsep). Since mitogen-activated protein kinases (MAPKs) may be downstream effectors of cPKC, we investigated here the involvement of the MAPKs of the ERK1/2, JNK1/2, and p38(MAPK) types in these deleterious effects...
December 2, 2016: Archives of Toxicology
https://www.readbyqxmd.com/read/27913741/aik1-a-mitogen-activated-protein-kinase-modulates-abscisic-acid-responses-through-the-mkk5-mpk6-kinase-cascade
#10
Kun Li, Fengbo Yang, Guozeng Zhang, Shufei Song, Yuan Li, Yuchen Miao, Dongtao Ren, Chun-Peng Song
The mitogen-activated protein kinase (MAPK) cascade is an evolutionarily conserved signal transduction module involved in transducing extracellular signals to the nucleus for appropriate cellular adjustment. This cascade essentially consists of three components, a MAPK kinase kinase (MAPKKK), a MAPK kinase (MAPKK) and a MAPK, connected to each other by the event of phosphorylation. Here we report the characterization of a MAPKKK, AIK1, which regulates abscisic acid (ABA) responses in Arabidopsis. T-DNA insertion mutants of AIK1 showed insensitivity to ABA in terms of both root growth and stomatal response...
December 2, 2016: Plant Physiology
https://www.readbyqxmd.com/read/27913520/myeloma-management-of-the-newly-diagnosed-high-risk-patient
#11
Angela Dispenzieri
Although there have been many definitions for high-risk (HR) myeloma, most recent consensus for classifying risk in patients with newly diagnosed multiple myeloma (NMM) comes from the International Myeloma Working Group. This recently published revised International Staging System includes del(17p) or t(4;14) by fluorescence in situ hybridization, β-2 microglobulin, albumin, and lactate dehydrogenase. These elements should be captured in all NMM patients. The optimal treatments for HR myeloma have not been fully worked out; therefore, these patients should be considered for clinical trials...
December 2, 2016: Hematology—the Education Program of the American Society of Hematology
https://www.readbyqxmd.com/read/27913473/first-line-therapy-for-young-patients-with-cll
#12
Nitin Jain, Susan O'Brien
A 61-year-old man with a history of chronic lymphocytic leukemia (CLL) presents with complaints of worsening fatigue and night sweats. He was diagnosed with CLL 3 years ago on routine blood count testing. He has no major medical comorbidities. On examination, he has several 2- to 3-cm lymph nodes in the cervical and axillary area. Spleen is palpable 5 cm below the costal margin. Blood counts show lymphocytosis with thrombocytopenia and anemia. Prognostic markers include deletion 13q by fluorescence in situ hybridization analysis and mutated IGHV You are asked by the hematology fellow you are supervising about the best treatment of this patient...
December 2, 2016: Hematology—the Education Program of the American Society of Hematology
https://www.readbyqxmd.com/read/27913288/autofluorescence-lifetime-variation-in-the-cuticle-of-the-bedbug-cimex-lectularius
#13
Klaus Reinhardt, Hans Georg Breunig, Karsten König
The decay time of the fluorescence of excited molecules, called fluorescence lifetime, can provide information about the cuticle composition additionally to widely used spectral characteristics. We compared autofluorescence lifetimes of different cuticle regions in the copulatory organ of females of the bedbug, Cimex lectularius. After two-photon excitation at 720 nm, regions recently characterised as being rich in resilin showed a longer bimodal distribution of the mean autofluorescence lifetime τm (tau-m) at 0...
November 29, 2016: Arthropod Structure & Development
https://www.readbyqxmd.com/read/27913171/effects-of-hydraulic-retention-time-hrt-on-denitrification-using-waste-activated-sludge-thermal-hydrolysis-liquid-and-acidogenic-liquid-as-carbon-sources
#14
Yiding Guo, Liang Guo, Mei Sun, Yangguo Zhao, Mengchun Gao, Zonglian She
Waste activated sludge (WAS) internal carbon source can efficiently and economically enhance denitrification, and hydraulic retention time (HRT) is one of the most important operational parameters for denitrification. The effects of HRT on denitrification were investigated with WAS thermal hydrolysis liquid and acidogenic liquid as carbon sources in this study. The optimal HRT was 12h for thermal hydrolysis liquid and 8h for acidogenic liquid, with NO3(-)-N removal efficiency of 91.0% and 97.6%, respectively...
November 15, 2016: Bioresource Technology
https://www.readbyqxmd.com/read/27913102/membrane-permeabilization-induced-by-triton-x-100-the-role-of-membrane-phase-state-and-edge-tension
#15
Bruno Mattei, Rafael B Lira, Katia R Perez, Karin A Riske
Detergents are widely used to solubilize and separate biomembrane components. It is therefore relevant to study and understand the mechanistic details underlying detergent-lipid interactions using biomimetic systems. Here, we have investigated in detail the process of membrane permeabilization and the nature of pores induced by sub-solubilizing concentrations of the detergent Triton X-100 (TX-100) in bilayers composed of palmitoyl oleoyl phosphatidylcholine (POPC), sphingomyelin (SM) and binary mixtures of these phospholipids with 30 mol% cholesterol (chol)...
November 29, 2016: Chemistry and Physics of Lipids
https://www.readbyqxmd.com/read/27913058/effect-of-urea-on-protein-ligand-association
#16
Lora Stepanian, Ikbae Son, Tigran V Chalikian
We combine experimental and theoretical approaches to investigate the influence of a cosolvent on a ligand-protein association event. We apply fluorescence measurements to determining the affinity of the inhibitor tri-N-acetylglucosamine [(GlcNAc)3] for lysozyme at urea concentrations ranging from 0 to 8M. Notwithstanding that, at room temperature and neutral pH, lysozyme retains its native conformation up to the solubility limit of urea, the affinity of (GlcNAc)3 for the protein steadily decreases as the concentration of urea increases...
November 25, 2016: Biophysical Chemistry
https://www.readbyqxmd.com/read/27912876/in-vitro-and-ex-vivo-activity-of-an-azadirachta-indica-a-juss-seed-kernel-extract-on-early-sporogonic-development-of-plasmodium-in-comparison-with-azadirachtin-a-its-most-abundant-constituent
#17
Nisha Dahiya, Giuseppina Chianese, Solomon Mequanente Abay, Orazio Taglialatela-Scafati, Fulvio Esposito, Giulio Lupidi, Massimo Bramucci, Luana Quassinti, George Christophides, Annette Habluetzel, Leonardo Lucantoni
BACKGROUND: NeemAzal(®) (NA) is a quantified extract from seed kernels of neem, Azadirachta indica A.Juss. (Meliaceae), with a wide spectrum of biological properties, classically ascribed to its limonoid content. NA contains several azadirachtins (A to L), azadirachtin A (AzaA) being its main constituent. AzaA has been shown to inhibit microgamete formation of the rodent malaria parasite Plasmodium berghei, and NA was found to completely inhibit the transmission of Plasmodium berghei to Anopheles stephensi mosquitoes when administered to gametocytemic mice at a corresponding AzaA dose of 50mg/kg before exposure to mosquitoes...
December 15, 2016: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/27912872/anti-allergic-potential-of-typhonium-blumei-inhibition-of-degranulation-via-suppression-of-pi3k-plc%C3%AE-2-phosphorylation-and-calcium-influx
#18
Michal Korinek, Kuan-Ming Chen, Yu-Han Jiang, Mohamed El-Shazly, Joel Stocker, Chon-Kit Chou, Tsong-Long Hwang, Yang-Chang Wu, Bing-Hung Chen, Fang-Rong Chang
BACKGROUND: Typhonium blumei Nicolson & Sivadasan (Araceae) is a traditional Chinese medicinal herb possessing detumescent, detoxifying, and anti-inflammatory activities. It is used in Taiwan as a folk medicine to treat cancer and inflammatory diseases. Typhonium blumei is usually not distinguished from Typhonium roxburghii Schott and they are commonly used interchangeably. PURPOSE: To evaluate and compare the anti-allergic and anti-inflammatory properties of T...
December 15, 2016: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/27912768/quasi-continuous-parallel-online-scattered-light-fluorescence-and-dissolved-oxygen-tension-measurement-combined-with-monitoring-of-the-oxygen-transfer-rate-in-each-well-of-a-shaken-microtiter-plate
#19
Tobias Ladner, Markus Held, David Flitsch, Mario Beckers, Jochen Büchs
BACKGROUND: Microtiter plates (MTP) are often applied as culture vessels in high-throughput screening programs. If online measuring techniques are available, MTPs can also be applied in the first steps of process development. For such small-scale bioreactors dipping probes are usually too large; therefore, optical measurements are often used. For example, the BioLector technology allows for the online monitoring of scattered light and fluorescence in each well of a continuously orbitally shaken MTP...
December 3, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27912186/recycling-oriented-comparison-of-mercury-distribution-in-new-and-spent-fluorescent-lamps-and-their-potential-risk
#20
Julia Hobohm, Oliver Krüger, Srotoswini Basu, Kerstin Kuchta, Sebastian van Wasen, Christian Adam
This study compares the mercury distribution in the vapor phase, the phosphor powder and the glass matrix of new and spent fluorescent lamps. The spent fluorescent lamps were obtained at the collection yards of a public waste management company in Hamburg, Germany. An innovative systematic sampling method is utilized to collect six spent and eight corresponding new, off-the-shelf fluorescent lamp samples. The efficiency of several acid digestion methods for the determination of the elemental composition was studied and elemental mass fractions of K, Na, Y, Ca, Ba, Eu, Al, Pb, Mg, Hg, and P were measured...
November 29, 2016: Chemosphere
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