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https://www.readbyqxmd.com/read/29198308/payload-hardware-and-experimental-protocol-development-to-enable-future-testing-of-the-effect-of-space-microgravity-on-the-resistance-to-gentamicin-of-uropathogenic-escherichia-coli-and-its-%C3%AF-s-deficient-mutant
#1
A C Matin, J-H Wang, Mimi Keyhan, Rachna Singh, Michael Benoit, Macarena P Parra, Michael R Padgen, Antonio J Ricco, Matthew Chin, Charlie R Friedericks, Tori N Chinn, Aaron Cohen, Michael B Henschke, Timothy V Snyder, Matthew P Lera, Shannon S Ross, Christina M Mayberry, Sungshin Choi, Diana T Wu, Ming X Tan, Travis D Boone, Christopher C Beasley, Matthew E Piccini, Stevan M Spremo
Human immune response is compromised and bacteria can become more antibiotic resistant in space microgravity (MG). We report that under low-shear modeled microgravity (LSMMG), stationary-phase uropathogenic Escherichia coli (UPEC) become more resistant to gentamicin (Gm), and that this increase is dependent on the presence of σs (a transcription regulator encoded by the rpoS gene). UPEC causes urinary tract infections (UTIs), reported to afflict astronauts; Gm is a standard treatment, so these findings could impact astronaut health...
November 2017: Life Sciences in Space Research
https://www.readbyqxmd.com/read/29196786/increased-heterologous-production-of-the-antitumoral-polyketide-mithramycin-a-by-engineered-streptomyces-lividans-tk24-strains
#2
Renata Novakova, Luz Elena Núñez, Dagmar Homerova, Renata Knirschova, Lubomira Feckova, Bronislava Rezuchova, Beatrica Sevcikova, Nuria Menéndez, Francisco Morís, Jesús Cortés, Jan Kormanec
Mithramycin A is an antitumor compound used for treatment of several types of cancer including chronic and acute myeloid leukemia, testicular carcinoma, hypercalcemia and Paget's disease. Selective modifications of this molecule by combinatorial biosynthesis and biocatalysis opened the possibility to produce mithramycin analogues with improved properties that are currently under preclinical development. The mithramycin A biosynthetic gene cluster from Streptomyces argillaceus ATCC12956 was cloned by transformation assisted recombination in Saccharomyces cerevisiae and heterologous expression in Streptomyces lividans TK24 was evaluated...
December 1, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29193817/isolation-and-purification-of-food-grade-c-phycocyanin-from-arthrospira-platensis-and-its-determination-in-confectionery-by-hplc-with-diode-array-detection
#3
Maria Kissoudi, Ioannis Sarakatsianos, Victoria Samanidou
C-Phycocyanin is the major phycobiliprotein in Arthrospira platensis, also known as Spirulina, which is a cyanobacterium used as a dietary supplement because of its powerful effects on body and brain. C-phycocyanin is a blue-colored accessory photosynthetic pigment with multiple applications in food industry as natural dye or additive, and in pharmaceuticals. This study presents a simple protocol for the extraction and purification of food-grade C-phycocyanin from Arthrospira platensis. The cell lysis of cyanobacterium was performed by sonication combined with repeated freezing and thawing cycles...
November 28, 2017: Journal of Separation Science
https://www.readbyqxmd.com/read/29188511/aav-gene-augmentation-therapy-for-crb1-associated-retinitis-pigmentosa
#4
C Henrique Alves, Jan Wijnholds
Mutations in the CRB1 gene account for around 10,000 persons with Leber congenital amaurosis (LCA) and 70,000 persons with retinitis pigmentosa (RP) worldwide. Therefore, the CRB1 gene is a key target in the fight against blindness. A proof-of-concept for an adeno-associated virus (AAV)-mediated CRB2 gene augmentation therapy for CRB1-RP was recently described. Preclinical studies using animal models such as knockout or mutant mice are crucial to obtain such proof-of-concept. In this chapter we describe a technique to deliver AAV vectors, into the murine retinas, via the subretinal route...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29182401/the-association-of-cataract-and-lens-epithelial-cell-apoptosis-in-patients-with-pseudoexfoliation-syndrome
#5
Pembe Oltulu, Refik Oltulu
PURPOSE: The aim of this study was to measure the density of apoptotic lens epithelial cells (LECs) and to determine its association with cataract formation in eyes with pseudoexfoliation syndrome (PEX). MATERIALS AND METHODS: Twenty-one eyes of 21 patients diagnosed with PEX without glaucoma (Group 1) and 22 eyes of 22 subjects without PEX (Group 2) were enrolled in this study. During cataract surgery, anterior capsule samples were obtained by the 5.5 mm continuous curvilinear capsulorhexis method...
November 28, 2017: Current Eye Research
https://www.readbyqxmd.com/read/29166269/selected-steps-of-swine-icsi-method-to-overcome-seasonal-effect-and-achieve-acceptable-early-embryonic-development-a-preliminary-study
#6
I A Tsakmakidis, M Tsantarliotou, I A Michos, A G Basioura, G Tsousis, C M Boscos
Porcine intra cytoplasmic sperm injection's (ICSI) efficacy by selected protocol steps was investigated. Three trials per year's period (hot, medium, cold) were carried out. Only large size follicles (6-8mm) were aspirated, brilliant cresyl blue (BCB) test was performed and only the BCB+ oocytes were in vitro maturated (40h) and involved to ICSI process. The presumptive embryos were in vitro cultured (15h). Raw boar semen and SpermCatch® as slowing medium were used. No differences were observed between periods regarding early embryonic development and maturation competence...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29161161/the-association-between-heavy-episodic-drinking-and-gender-orientation-among-u-s-college-students-the-significance-of-masculinity
#7
Robert L Peralta, Marcella L Mulhollem, Courtney Blue, Breanna C Stewart
BACKGROUND: Heavy episodic drinking (HED) remains a public health concern among college students. Sex differences are routinely reported in the literature although some evidence of convergence in drinking patterns has been observed. The association between sex and gender-orientation in HED remains unclear because sex and gender are often conflated. OBJECTIVE: We examine the intersection of sex, gender-orientation and HED to determine if gender-orientation alone and/or in conjunction with sex play a role in HED among college students...
November 21, 2017: Substance Use & Misuse
https://www.readbyqxmd.com/read/29155787/synthesis-of-core-shell-lanthanide-doped-upconversion-nanocrystals-for-cellular-applications
#8
Xiangzhao Ai, Linna Lyu, Jing Mu, Ming Hu, Zhimin Wang, Bengang Xing
Lanthanide-doped upconversion nanocrystals (UCNs) have attracted much attention in recent years based on their promising and controllable optical properties, which allow for the absorption of near-infrared (NIR) light and can subsequently convert it into multiplexed emissions that span over a broad range of regions from the UV to the visible to the NIR. This article presents detailed experimental procedures for high-temperature co-precipitation synthesis of core-shell UCNs that incorporate different lanthanide ions into nanocrystals for efficiently converting deep-tissue penetrable NIR excitation (808 nm) into a strong blue emission at 480 nm...
November 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29154914/how-to-make-usage-of-the-standardized-euroflow-8-color-protocols-possible-for-instruments-of-different-manufacturers
#9
Michaela Nováková, Hana Glier, Naděžda Brdičková, Marcela Vlková, Ana Helena Santos, Margarida Lima, Mikael Roussel, Juan Flores-Montero, Tomasz Szczepanski, Sebastian Böttcher, Vincent H J van der Velden, Paula Fernandez, Ester Mejstříková, Leire Burgos, Bruno Paiva, Jacques J M van Dongen, Alberto Orfao, Tomáš Kalina
A critical component of the EuroFlow standardization of leukemia/lymphoma immunophenotyping is instrument setup. Initially, the EuroFlow consortium developed a step-by-step standard operating protocol for instrument setup of ≥8-color flow cytometers that were available in 2006, when the EuroFlow activities started. Currently, there are 14 instruments from 9 manufacturers capable of 3-laser excitation and ≥8 color measurements. The specific adaptations required in the instrument set-up to enable them to acquire the standardized 8-color EuroFlow protocols are described here...
November 15, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29152022/proteomic-approach-for-extracting-cytoplasmic-proteins-from-streptococcus-sanguinis-using-mass-spectrometry
#10
Fadi El-Rami, Kristina Nelson, Ping Xu
Streptococcus sanguinis is a commensal and early colonizer of oral cavity as well as an opportunistic pathogen of infectious endocarditis. Extracting the soluble proteome of this bacterium provides deep insights about the physiological dynamic changes under different growth and stress conditions, thus defining "proteomic signatures" as targets for therapeutic intervention. In this protocol, we describe an experimentally verified approach to extract maximal cytoplasmic proteins from Streptococcus sanguinis SK36 strain...
2017: Journal of molecular biology research
https://www.readbyqxmd.com/read/29150736/effect-of-erythrocyte-sperm-separation-medium-on-nuclear-acrosomal-and-membrane-maturity-parameters-in-human-sperm
#11
Bikem Soygur, Soner Celik, Ciler Celik-Ozenci, Leyla Sati
PURPOSE: The purpose of this study is to investigate whether erythrocyte-sperm separation medium (ESSM) has effects on human sperm motility, morphology, viability, membrane maturity, acrosome integrity, and nuclear attributes before and after cryopreservation. METHODS: Semen samples from normozoospermic (n = 36) and oligozoospermic (n = 9) patients were analyzed. Samples from the same patient were divided into three aliquots: group 1 and group 2 were resuspended in sperm washing media and ESSM, respectively...
November 18, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/29135249/photoreductive-removal-of-o-benzyl-groups-from-oxyarene-n-heterocycles-assisted-by-o-pyridine-pyridone-tautomerism
#12
Aleksandar R Todorov, Tom Wirtanen, Juho Helaja
Facile photoreductive protocols have been developed to remove benzyl O-protective groups from oxyarene N-heterocycles at positions capable for 2-/4-O-pyridine - 2-/4-pyridone tautomerism. Blue light irradiation, a [Ru] or [Ir] photocatalyst and ascorbic acid in a water-acetonitrile solution debenzylates a variety of aryl N-heterocycles cleanly and selectively. Ascorbic acid has two functions in the reaction. On the one hand, it protonates the N-heterocycles that reduces their reduction potentials notably and on the hand it acts as a sacrificial reductant...
November 14, 2017: Journal of Organic Chemistry
https://www.readbyqxmd.com/read/29134509/selection-of-an-effective-indicator-for-rapid-detection-of-microorganisms-producing-%C3%AE-polyglutamic-acid-and-its-biosynthesis-under-submerged-fermentation-conditions-using-bacillus-methylotrophicus
#13
Poonam Mishra Chatterjee, Silpi Datta, Deepika Pandey Tiwari, Ritu Raval, Ashok Kumar Dubey
γ-Polyglutamic acid (γ-PGA) is a biosynthetic outcome of glutamic acid polymerization by microbes. In the current study, we have isolated Bacillus methylotrophicus on solid differential media containing methylene blue. This is the first report mentioning the use of methylene blue to distinguish the monomeric and polymeric form of glutamic acid in the liquid medium using UV-Vis spectrophotometer. Our method can simplify the analytical process of γ-PGA confirmation using the aforementioned studies. This screening protocol is sensitive to the detection of γ-PGA quantities as low as 3 μg/mL; thus, the potent producers can be effectively screened...
November 13, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29133761/reduction-on-off-responses-of-electroretinogram-in-monkeys-with-long-term-high-intraocular-pressure
#14
Ke-Gao Liu, Xiao-Yan Peng, Zheng Zhang, Hua Sun, Di-Ya Yang, Ning-Li Wang
BACKGROUND: There are ON- and OFF-pathways in the normal vertebrate retina. Short- and long-flash electroretinogram (ERG) are suitable methods to observe the function of ON- and OFF-pathways in vivo, respectively. It is clear that high intraocular pressure (IOP) might cause dysfunction of cone-dominated photopic negative response (PhNR) in monkeys with high IOP in ON-pathway. However, whether cone-dominated OFF-responses are also affected is less known. The aim of this study was to observe photopic OFF-responses of ERG in monkeys with high IOP...
November 20, 2017: Chinese Medical Journal
https://www.readbyqxmd.com/read/29126958/effects-of-led-based-photodynamic-therapy-using-red-and-blue-lights-with-natural-hydrophobic-photosensitizers-on-human-glioma-cell-line
#15
Zahra Jamali, Sedigheh Marjaneh Hejazi, Seyed Mohsen Ebrahimi, Hemen Moradi-Sardareh, Maliheh Paknejad
Photodynamic therapy (PDT) has received high attention in cancer treatment due to its minimal side effects, specific cancer-targeting, non-invasion and low cost. It utilizes a specific group of anti-cancer drugs called photosensitizers (PS), which can be only activated under a certain wavelength light illumination and kills cancer cells. To screen the potential of PS and setup of PDT treatment protocol, it is essential to assess the PDT efficacy in vitro. In this study, a light-emitting diode- (LED-) based illumination system at two wavelengths (red & blue) with homogeneous and stable irradiation, and constant temperature conditions in 96-well plates was provided...
November 7, 2017: Photodiagnosis and Photodynamic Therapy
https://www.readbyqxmd.com/read/29122403/validation-of-simple-and-cost-effective-stains-to-assess-acrosomal-status-dna-damage-and-mitochondrial-activity-in-rooster-spermatozoa
#16
Bruno R Rui, Daniel S R Angrimani, João Diego A Losano, Luana de Cássia Bicudo, Marcílio Nichi, Ricardo J G Pereira
Several methods have been developed to evaluate spermatozoa function in birds but many of these are sometimes complicated, costly and not applicable to field studies (i.e., performed within poultry breeding facilities). The objective was, therefore, to validate efficient, practical and inexpensive procedures to determine DNA fragmentation, acrosomal integrity, and mitochondrial activity in poultry spermatozoa. Initially, ejaculates were individually diluted and divided into control (4°C, 4h) and UV-irradiated aliquots (room temperature, 4h), and then samples containing different percentages of DNA-damaged spermatozoa (0%, 25%, 50%, 75% and 100%) were subjected to Toluidine Blue (TB) and Sperm Chromatin Dispersion assessments (SCD)...
December 2017: Animal Reproduction Science
https://www.readbyqxmd.com/read/29119417/effect-of-methylene-blue-mediated-antimicrobial-photodynamic-therapy-on-dentin-caries-microcosms
#17
Daniela Alejandra Cusicanqui Méndez, Eliezer Gutierrez, Evandro José Dionísio, Thaís Marchini Oliveira, Marília Afonso Rabelo Buzalaf, Daniela Rios, Maria Aparecida Andrade Moreira Machado, Thiago Cruvinel
Antimicrobial photodynamic therapy (aPDT) has been proposed as an adjuvant treatment of dental caries, although there are no well-defined protocols to its clinical application. This study aimed to evaluate the influence of aPDT on the viability of microorganisms, vitality of biofilms, and lactic acid production of dentin caries microcosms. Biofilms were grown on bovine dentin discs in anaerobic conditions at 37 °C for 5 days, inoculating infected carious dentin in modified McBain medium plus 1% sucrose. The biofilms were treated by the combination of deionized water or 100 mg L(-1) methylene blue (MB) with 0, 37...
November 8, 2017: Lasers in Medical Science
https://www.readbyqxmd.com/read/29113281/direct-effects-of-phenformin-on-metabolism-bioenergetics-and-viability-of-sh-sy5y-neuroblastoma-cells
#18
Fintan Geoghegan, Naomi Chadderton, G Jane Farrar, Daniela M Zisterer, Richard K Porter
Phenformin, a member of the biguanides class of drugs, has been reported to be efficacious in cancer treatment. The focus of the current study was to establish whether there were direct effects of phenformin on the metabolism and bioenergetics of neuroblastoma SH-SY5Y cancer cells. Cell viability was assessed using the alamar blue assay, flow cytometry analysis using propidium iodide and annexin V stain and poly (ADP-ribose) polymerase analysis. Cellular and mitochondrial oxygen consumption was determined using a Seahorse Bioscience Flux analyser and an Oroboros Oxygraph respirometer...
November 2017: Oncology Letters
https://www.readbyqxmd.com/read/29097290/in-vitro-evaluation-of-methylene-blue-removal-from-root-canal-after-photodynamic-therapy
#19
Karen Müller Ramalho, Sandra Ribeiro Cunha, Eric Mayer-Santos, Carlos de Paula Eduardo, Patricia Moreira de Freitas, Ana Cecilia Correa Aranha, Cacio Moura-Netto
In Endodontics, photosensitizers' such as methylene blue and toluidine blue have been used in photodynamic therapy due to their positive results. However, they can stain the dentin from the root canal after Photodynamic Therapy (PDT). The present in vitro study aimed to evaluate different stain removal protocols from root canal after PDT using methylene blue (MB) dye. After mechanical preparation of the root canal of 40 uni-radicular human teeth, PDT was performed using 0,01% MB with parameters of 3minutes of pre-irradiation and a diode laser irradiation emitting at 660nm, 40mW, 4minutes, 9...
October 30, 2017: Photodiagnosis and Photodynamic Therapy
https://www.readbyqxmd.com/read/29078067/bluephage-a-rapid-method-for-the-detection-of-somatic-coliphages-used-as-indicators-of-fecal-pollution-in-water
#20
M Muniesa, E Ballesté, L Imamovic, M Pascual-Benito, D Toribio-Avedillo, F Lucena, A R Blanch, J Jofre
The use of somatic coliphages as indicators of fecal and viral pollution in water and food has great potential due to the reliability, reproducibility, speed and cost effectiveness of methods for their detection. Indeed, several countries already use this approach in their water management policies. Although standardized protocols for somatic coliphage detection are available, user-friendly commercial kits would facilitate their routine implementation in laboratories. The new method presented here allows detection of up to 1 somatic coliphage in under 3...
October 16, 2017: Water Research
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