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https://www.readbyqxmd.com/read/27906168/immunofluorescent-antibody-staining-of-intact-drosophila-larvae
#1
Laurina Manning, Chris Q Doe
Antibody staining is a vital technique for studying the development of many model organisms, including Drosophila. Reliable protocols have long been available for antibody staining of Drosophila whole-mount embryos and dissected larvae. By contrast, methods for staining whole larvae have rarely been reported, are unreliable, and fail to work on large third-instar larvae. This has become a major limitation to understanding the role of multitissue interactions such as neural circuit formation and cell metastasis...
January 2017: Nature Protocols
https://www.readbyqxmd.com/read/27899273/sandtraps-are-efficient-scalable-and-mild-systems-for-harvesting-washing-and-concentrating-cells
#2
Danny Frauenstein, Katja Seidel, Lorenz Adrian
Microbial cells vary widely in size, specific density, shearing resistance, oxygen sensitivity and abundance so that differential harvesting and washing procedures are needed to efficiently recover cells from dilute suspensions. We here describe a mild, simple, variable and cost-efficient method to harvest cells on columns packed with silica beads. The method collects and concentrates 40-98% of the cells preserving enzymatic activity and cell viability. The method can be applied for strictly anaerobic microorganisms, is scalable to different culture volumes and can be multiplexed in standardized systems...
November 26, 2016: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/27898431/improving-outcome-of-trauma-patients-by-implementing-patient-blood-management
#3
Christoph Füllenbach, Kai Zacharowski, Patrick Meybohm
PURPOSE OF REVIEW: Patient blood management aims to improve patient outcome and safety by reducing the number of unnecessary red blood cell transfusions and vitalizing patient-specific anemia reserves. While this is increasingly recognized as best clinical practice in elective surgery, the implementation in the setting of trauma is restrained because of typically nonelective (emergency) surgery and, in specific circumstances, allogeneic blood transfusions as life-saving therapy. RECENT FINDINGS: Viscoelastic diagnostics allow a precise identification of trauma-induced coagulopathy...
November 24, 2016: Current Opinion in Anaesthesiology
https://www.readbyqxmd.com/read/27895534/an-automated-perifusion-system-for-modifying-cell-culture-conditions-over-time
#4
Nicholas B Whitticar, Elisha W Strahler, Parthiban Rajan, Savas Kaya, Craig S Nunemaker
BACKGROUND: Cells are continuously exposed to changes in their environment. Endocrine systems, in particular, communicate by rhythms and feedback loops. In this study, we developed an automated system to produce such conditions for cultured cells in a precisely timed manner. We utilized a programmable pair of syringe pumps for inflow and a peristaltic pump for outflow to create rhythmic pulses at 5-min intervals in solutions that mimic the endogenous patterns of insulin produced by pancreatic islets as a test case...
2016: Biological Procedures Online
https://www.readbyqxmd.com/read/27891466/an-in-vitro-evaluation-of-retention-colonization-and-penetration-of-commonly-used-denture-lining-materials-by-candida-albicans
#5
Sushma Krishnamurthy, Rajendra B Hallikerimath
INTRODUCTION: Colonization of the surface by Candida albicans and related Candida species is one of the major concerns of denture lining materials. AIM: We evaluated retention, colonization and penetration of the four denture lining materials namely Molloplast B, Permaflex, GC Soft Liner and Ufi Gel Hard C by Candida albicans. MATERIAL AND METHODS: a) Evaluation of retention: Five test discs of each material with smooth surface on one side and rough on the other were prepared and surface roughness (Ra) was measured with profilometer...
October 2016: Journal of Clinical and Diagnostic Research: JCDR
https://www.readbyqxmd.com/read/27889603/two-symbiotic-bacteria-of-the-entomopathogenic-nematode-heterorhabditis-spp-against-galleria-mellonella
#6
Chunli Liao, Along Gao, Bingbing Li, Mengjun Wang, Linna Shan
The entomopathogenic nematode Heterorhabditis spp. is considered a promising agent in the biocontrol of injurious insects of agriculture. However, different symbiotic bacteria associated with the nematode usually have different specificity and virulence toward their own host. In this study, two symbiotic bacteria, LY2W and NK, were isolated from the intestinal canals of two entomopathogenic nematode Heterorhabditis megidis 90 (PDSj1 and PDSj2) from Galleria mellonela, separately. To determine their species classification, we carried out some investigations on morphology, culture, biochemistry, especially 16S rDNA sequence analyses...
November 23, 2016: Toxicon: Official Journal of the International Society on Toxinology
https://www.readbyqxmd.com/read/27888164/optimizing-zn-porphyrin-based-photosensitizers-for-efficient-antibacterial-photodynamic-therapy
#7
Khazna Alenezi, Artak Tovmasyan, Ines Batinic-Haberle, Ludmil T Benov
INTRODUCTION: Efficient photodynamic inactivation of microbes requires highly efficient photosensitizers which kill microbial cells, but spare host tissues. One way to achieve such selectivity is to use photosensitizers that are rapidly taken up by microbes and, when applied at low concentrations, efficiently kill them after a short illumination. Design of such photosensitizers requires insight into molecular properties which are critical for antimicrobial photo-efficiency. This study explores the contribution of molecular shape and exposure of charges, to the antimicrobial activity of tetra-cationic Zn porphyrin-based photosensitizers...
November 22, 2016: Photodiagnosis and Photodynamic Therapy
https://www.readbyqxmd.com/read/27884932/drosophila-wash-is-required-for-integrin-mediated-cell-adhesion-cell-motility-and-lysosomal-neutralization
#8
Benedikt M Nagel, Meike Bechtold, Luis Garcia Rodriguez, Sven Bogdan
The Wiskott-Aldrich Syndrome Protein and SCAR Homologue (WASH) is a conserved actin nucleation promoting factor controlling Arp2/3 complex activity in endosomal sorting and recycling. Previous studies have identified WASH as an essential regulator in Drosophila development. Here, we show that homozygous wash mutant flies are viable and fertile. We demonstrate that Drosophila WASH has conserved functions in integrin receptor recycling and lysosome neutralization. WASH generates actin patches on endosomes and lysosomes mediating both functions...
November 24, 2016: Journal of Cell Science
https://www.readbyqxmd.com/read/27878040/effect-of-contrast-media-on-urinary-cytopathology-specimens
#9
Sebastian Frees, Samir Bidnur, Michael Metcalfe, Peter Raven, Claudia Chavez-Munoz, Igor Moskalev, Ladan Fazli, Alan So
INTRODUCTION: Urological dogma dictates that washings collected from the urinary tract for cytological assessment must be performed without interference from contrast agents that may alter cellular integrity and diagnostic interpretation. In practice, the initial contrast used to outline the upper tracts is commonly discarded with subsequent saline washings sent for cytology. We hypothesize that contrast washings do not affect the morphology of urothelial carcinoma cells or the integrity of cytology interpretation...
August 2016: Canadian Urological Association Journal, Journal de L'Association des Urologues du Canada
https://www.readbyqxmd.com/read/27877882/focal-calcium-monitoring-with-targeted-nanosensors-at-the-cytosolic-side-of-endoplasmic-reticulum
#10
Yanyan Hou, Satoshi Arai, Yoshiaki Takei, Atsushi Murata, Shinji Takeoka, Madoka Suzuki
Ca(2+) distribution is spatially and temporally non-uniform inside cells due to cellular compartmentalization. However, Ca(2+) sensing with small organic dyes, such as fura-2 and fluo-4, has been practically applied at a single cell level where the averaged signal from freely diffusing dye molecules is acquired. In this study, we aimed to target azide-functionalized fura-2 (N3-fura-2) to a specific site of subcellular compartments to realize focal Ca(2+) sensing. Using scAVD (single-chain avidin)-biotin interaction and a copper-free click reaction system, we linked N3-fura-2 to specifically-targeted scAVD protein fused with a red fluorescent protein mCherry, so that Ca(2+) sensors conjugated with four N3-fura-2 dyes with dibenzocyclooctyne (DBCO)-PEG4-biotin as a linker were generated at subcellular compartments in living cells...
2016: Science and Technology of Advanced Materials
https://www.readbyqxmd.com/read/27877879/a-stem-less-probe-using-spontaneous-pairing-between-cy3-and-quencher-for-rna-detection
#11
Hiromu Kashida, Kazuhiro Morimoto, Hiroyuki Asanuma
We herein report a stem-less probe for the detection of RNA that depends on pairing between Cy3 and nitro methyl red. In our design, two Cy3 residues and two nitro methyl red residues were introduced into an oligonucleotide. In the absence of the target, these dyes formed a complex, and emission of Cy3 was efficiently quenched. Hybridization with the target RNA disrupted this interaction and resulted in Cy3 emission. Under optimized conditions, the signal to background ratio was as high as 180. We demonstrated specific detection of target RNA in cells using a wash-free FISH protocol...
2016: Science and Technology of Advanced Materials
https://www.readbyqxmd.com/read/27875751/all-in-one-dual-aptasensor-capable-of-rapidly-quantifying-carcinoembryonic-antigen
#12
Harriet Khang, Kelly Cho, Stephanie Chong, Ji Hoon Lee
Using a dual DNA aptamer (CEA aptamer linked to hemin aptamer), capable of rapidly capturing carcinoembryonic antigen (CEA) and hemin, an all-in-one dual-aptasensor with 1,1'-oxalyldiimidazole (ODI) chemiluminescence detection was developed for the early diagnosis of human cancer. CEA and hemin competitively bound with the dual DNA aptamer while the mixture in a detection cell was incubated for 30min at room temperature. When Amplex Red and H2O2 were added in the detection cell after the incubation, the yield of resorufin formed from the reaction Amplex Red and H2O2 depended on the concentration of HRP-mimicking G-quardruplex DNAzyme formed from the binding interaction between hemin and the dual DNA aptamer...
November 16, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27875619/automated-leukocyte-processing-by-microfluidic-deterministic-lateral-displacement
#13
Curt I Civin, Tony Ward, Alison M Skelley, Khushroo Gandhi, Zendra Peilun Lee, Christopher R Dosier, Joseph L D'Silva, Yu Chen, MinJung Kim, James Moynihan, Xiaochun Chen, Lee Aurich, Sergei Gulnik, George C Brittain, Diether J Recktenwald, Robert H Austin, James C Sturm
We previously developed a Deterministic Lateral Displacement (DLD) microfluidic method in silicon to separate cells of various sizes from blood (Davis et al., Proc Natl Acad Sci 2006;103:14779-14784; Huang et al., Science 2004;304:987-990). Here, we present the reduction-to-practice of this technology with a commercially produced, high precision plastic microfluidic chip-based device designed for automated preparation of human leukocytes (white blood cells; WBCs) for flow cytometry, without centrifugation or manual handling of samples...
November 22, 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/27872531/positive-peritoneal-lavage-cytology-implications-for-staging-and-management-of-gastric-cancer
#14
Suman B Koganti, Satish Boddepalli, Muralidhar Nambada, Venu Madhav Thumma, Bheerappa Nagari, R A Sastry
The survival rates of gastric cancer patients with cytology-positive peritoneal lavage fluid without macroscopic dissemination (CY+/P-) is the same as that of patients with overt peritoneal metastasis.The 5-year survival rate of such patients is only 2%. The current study aims to highlight its significance in the staging of gastrointestinal malignancies and its implications for patient care. Prospective nonrandom analysis of peritoneal wash cytology in patients with gastrointestinal malignancies was conducted in the department of Surgical Gastroenterology, Nizams Institute of Medical Sciences, Hyderabad from January 2012 to June 2013...
December 2016: Indian Journal of Surgical Oncology
https://www.readbyqxmd.com/read/27866824/detection-of-her2-overexpressing-cancer-cells-using-keyhole-shaped-chamber-array-employing-a-magnetic-droplet-handling-system
#15
Mina Okochi, Shinji Koike, Masayoshi Tanaka, Hiroyuki Honda
An on-chip gene expression analysis compartmentalized in droplets was developed for detection of cancer cells at a single-cell level. The chip consists of a keyhole-shaped reaction chamber with hydrophobic modification employing a magnetic bead-droplet-handling system with a gate for bead separation. Using three kinds of water-based droplets in oil, a droplet with sample cells, a lysis buffer with magnetic beads, and RT-PCR buffer, parallel magnetic manipulation and fusion of droplets were performed using a magnet-handling device containing small external magnet patterns in an array...
November 11, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27865110/specifically-and-wash-free-labeling-of-snap-tag-fused-proteins-with-a-hybrid-sensor-to-monitor-local-micro-viscosity
#16
Chao Wang, Xinbo Song, Lingcheng Chen, Yi Xiao
Viscosity, as one of the major factors of intracellular microenvironment, influences the function of proteins. To detect local micro-viscosity of a protein, it is a precondition to apply a viscosity sensor for specifically target to proteins. However, all the reported small-molecule probes are just suitable for sensing/imaging of macro-viscosity in biological fluids of entire cells or organelles. To this end, we developed a hybrid sensor BDP-V BG by connecting a viscosity-sensitive boron-dipyrromethene (BODIPY) molecular rotor (BDP-V) to O(6)-benzylguanine (BG) for specific detection of local micro-viscosity of SNAP-tag fused proteins...
November 10, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27864978/reduction-of-biological-response-modifiers-in-the-supernatant-of-washed-paediatric-red-blood-cells
#17
Y S Loh, S Tan, M Kwok, M J Stark, D C Marks
BACKGROUND: Washing of red blood cells (RBC) can reduce unwanted biological response modifiers (BRMs) that can mediate transfusion complications in infants. The aim of this study was to examine the in vitro quality and the changes in BRMs following washing in paediatric RBC units. MATERIALS AND METHODS: A pool and split design was used to prepare RBC (either 1 or 4 days old; n = 26 pairs). One unit was washed with 0·9% saline by centrifugation and then resuspended in SAG-M, while the other remained unwashed...
November 2016: Vox Sanguinis
https://www.readbyqxmd.com/read/27864173/salad-leaf-juices-enhance-salmonella-growth-fresh-produce-colonisation-and-virulence
#18
Giannis Koukkidis, Richard Haigh, Natalie Allcock, Suzanne Jordan, Primrose Freestone
: We show in this report that traces of juices released from salad leaves as they became damaged can significantly enhance Salmonella enterica salad leaf colonisation. Salad juices in water increased Salmonella growth by 110% over the un-supplemented control, and in host-like serum based media by more than 2400-fold over controls. In serum based media salad juices induced growth of Salmonella via provision of Fe from transferrin, and siderophore production was found to be integral to the growth induction process...
November 18, 2016: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/27861545/glucocorticoid-induces-incoordination-between-glutamatergic-and-gabaergic-neurons-in-the-amygdala
#19
Guang-Yan Wang, Zhao-Ming Zhu, Shan Cui, Jin-Hui Wang
BACKGROUND: Stressful life leads to mood disorders. Chronic mild stress is presumably major etiology for depression, and acute severe stress leads to anxiety. These stressful situations may impair hypothalamus-pituitary-adrenal axis and in turn induce synapse dysfunction. However, it remains elusive how the stress hormones mess up subcellular compartments and interactions between excitatory and inhibitory neurons, which we have investigated in mouse amygdala, a structure related to emotional states...
2016: PloS One
https://www.readbyqxmd.com/read/27855889/bioavailability-of-angiotensin-i-converting-enzyme-ace-inhibitory-peptides-derived-from-virgibacillus-halodenitrificans-sk1-3-7-proteinases-hydrolyzed-tilapia-muscle-proteins
#20
Tidarat Toopcham, Jurriaan J Mes, Harry J Wichers, Sittiruk Roytrakul, Jirawat Yongsawatdigul
The angiotensin I-converting enzyme (ACE) inhibitory activity of protein hydrolysates from tilapia muscle fractions, namely mince (M), washed mince (WM), and sarcoplasmic protein (SP), were investigated. Each fraction was hydrolyzed by Virgibacillus halodenitrificans SK1-3-7 proteinases for up to 24h. After 8h of hydrolysis, the M hydrolysate (48% degree of hydrolysis (DH)) showed the highest ACE inhibitory activity, with an IC50 value of 0.54mg/ml, while the SP hydrolysate exhibited the lowest DH and ACE inhibition...
April 1, 2017: Food Chemistry
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