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Gili Dardikman, Yoav N Nygate, Itay Barnea, Nir A Turko, Gyanendra Singh, Barham Javidi, Natan T Shaked
We suggest a new multimodal imaging technique for quantitatively measuring the integral (thickness-average) refractive index of the nuclei of live biological cells in suspension. For this aim, we combined quantitative phase microscopy with simultaneous 2-D fluorescence microscopy. We used 2-D fluorescence microscopy to localize the nucleus inside the quantitative phase map of the cell, as well as for measuring the nucleus radii. As verified offline by both 3-D confocal fluorescence microscopy and 2-D fluorescence microscopy while rotating the cells during flow, the nucleus of cells in suspension that are not during division can be assumed to be an ellipsoid...
March 1, 2018: Biomedical Optics Express
Joshua S Elmore, Michael H Baumann
Naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-018) is a synthetic compound found in psychoactive "spice" products that activates cannabinoid receptors. Preclinical evidence suggests that exposure to synthetic cannabinoids increases 5-HT2A/2C receptor function in the brain, an effect which might contribute to psychotic symptoms. Here, we hypothesized that repeated exposures to JWH-018 would enhance behavioral responsiveness to the 5-HT2A/2C receptor agonist DOI. Male Sprague-Dawley rats fitted with subcutaneously (sc) temperature transponders received daily injections of JWH-018 (1...
2018: Frontiers in Psychiatry
Claudia Miranda, Zenia Pérez-Rodríguez, Rosario Hernández-Armengol, Yaidel Quiñones-García, Tania Betancourt-Purón, Miguel Ángel Cabrera-Pérez
The aim of the present study is to contribute to the scientific characterization of sildenafil citrate according to the Biopharmaceutics Classification System, following the World Health Organization (WHO) guidelines for biowaivers. The solubility and intestinal permeability data of sildenafil citrate were collected from literature; however, the experimental solubility studies are inconclusive and its "high permeability" suggests an API in the borderline of BCS Class I and Class II. The pH-solubility profile was determined using the saturation shake-flask method over the pH range of 1...
March 12, 2018: AAPS PharmSciTech
Hua Wang, Wenxi Zhou, Hua Li, Rie Bu
The cyclodextrin glycosyltransferase (CGTase) was used to catalyze the conversion of starch into cyclodextrins (CD) in industry. Improving the activity of CGTase to produce more CD with relative low cost is intensely interesting and has drawn wide attention. Amino acid mutation of His167 into Cys significantly enhanced β-CGTase activity; however, optimization of culture conditions for β-CGTase-H167C remains unclear. To determine this, the medium and culture conditions for β-CGTase-H167C were optimized with response surface methodology...
March 2018: 3 Biotech
Yujia Zhao, Jingjing Fan, Chen Wang, Xudong Feng, Chun Li
Oleanolic acid is a plant-derived pentacyclic triterpenoid compound with various biological activities. Recently, biosynthesis of oleanolic acid in microbes has been demonstrated as a promising and green way, but the production is too low for industrialization. To improve oleanolic acid production, this study constructed a novel pathway for biosynthesis of oleanolic acid in Saccharomyces cerevisiae by improving the pairing efficiency between cytochrome P450 monooxygenase and reductase. Furthermore, to improve the transcriptional efficiency of heterologous genes, the cellular galactose regulatory network was reconstructed by knocking out galactose metabolic genes GAL80 and GAL1...
February 23, 2018: Bioresource Technology
Kelly J Gambino-Shirley, Adiam Tesfai, Colin A Schwensohn, Cindy Burnett, Lori Smith, Jennifer M Wagner, Dana Eikmeier, Kirk Smith, Jolianne P Stone, Dawn Updike, Jonas Hines, Lauren N Shade, Beth Tolar, Tong-Jen Fu, Stelios Viazis, Sharon L Seelman, Kathryn Blackshear, Matthew E Wise, Karen P Neil
Background: Nontyphoidal Salmonella is the leading cause of bacterial gastroenteritis in the United States. Meal replacement products containing raw and 'superfood' ingredients have gained increasing popularity among consumers in recent years. In January 2016, we investigated a multistate outbreak of infections with a novel strain of Salmonella Virchow. Methods: Cases were defined using molecular subtyping procedures. Commonly reported exposures were compared with responses from healthy people interviewed in the 2006-2007 FoodNet Population Survey...
March 7, 2018: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
Quan Li, Yuhang Zhang, Du Dechao, Yu Yanfei, Wei Zhang
Streptococcus suis, an important swine pathogen and a major zoonotic agent, is responsible for severe financial losses in the global swine industry. Although a multitude of virulence factors have been reported, the pathogenesis of S. suis infections remains poorly understood. In our previous work, we identified a potential virulence-associated protein, named PnuC, unique to virulent strains of S. suis serotype 2 (S. suis 2). To investigate the functions of PnuC, the pnuC gene deletion mutant (ΔpnuC) was constructed in S...
March 2018: Veterinary Microbiology
Wen Zhu, Guihua Gong, Jie Pan, Shu Han, Wei Zhang, Youjia Hu, Liping Xie
Human serum albumin (HSA) has been extensively used in a series of clinical care settings for nearly seven decades. However, the broad application of this protein is seriously limited by its short supply. In this work, the codon sequence of HSA was cloned under the control of the alcohol oxidase 1 promoter (AOX1) and expressed as a secretory protein in Pichia pastoris. A recombinant strain displaying the highest HSA yield was selected by screening for resistance to the highest concentration of antibiotic G418...
March 5, 2018: Protein Expression and Purification
Sarah L Irons, Adam C Chambers, Olga Lissina, Linda A King, Robert D Possee
Baculovirus expression systems are well established as an easy and reliable way to produce high quality recombinant proteins. Baculoviruses can also be used to transduce mammalian cells, termed 'BacMam', with considerable potential in biomedical applications. This chapter explains the process of making a recombinant baculovirus, encompassing production of a recombinant virus by homologous recombination in insect cells, followed by amplification and titration of the virus-all steps needed before commencing gene expression and protein production...
February 21, 2018: Current Protocols in Protein Science
Reshma John, P P Shajitha, Aneesha Devassy, Linu Mathew
20-Hydroxyecdysone is one of the most common ecdysteroids in plants with potential therapeutic applications. In this study, cell suspension cultures of Achyranthes aspera were raised in shake flasks to investigate the production of 20-hydroxyecdysone. The quantification and characterization of 20-hydroxyecdysone in the cultures were done by High performance liquid chromatography (HPLC) and Liquid Chromatography-quadrupole time-of- flight mass spectrometry (LC-Q-TOF) analyses. For raising the suspension, calli initiated from in vitro grown leaf explants were cultured in liquid Murashige and Skoog (MS) medium augmented with combinations of 2, 4-dichlorophenoxyacetic acid (1 mg L-1 ) and α-naphthaleneacetic acid (1 mg L-1 )...
March 2018: Physiology and Molecular Biology of Plants: An International Journal of Functional Plant Biology
Marianna Yurkiv, Olena Kurylenko, Roksolana Vasylyshyn, Kostyantyn Dmytruk, Patrick Fickers, Andriy Sibirny
Glutathione is the most abundant cellular thiol and the low molecular weight peptide present in cells. The methylotrophic yeast Ogataea (Hansenula) polymorpha is considered as a promising cell factory for the synthesis of glutathione. In this study, а competitive O. polymorpha glutathione producer was constructed by overexpression of the GSH2 gene, encoding γ-glutamylcysteine synthetase, the first enzyme involved in glutathione biosynthesis, and the MET4 gene coding for central regulator of sulfur metabolism...
March 4, 2018: FEMS Yeast Research
Geoffrey David Debelle, Sabine Maguire, Patrick Watts, Rosa Nieto Hernandez, Alison Mary Kemp
The Swedish Agency for Health Technology Assessment and Assessment of Social Services (SBU) has recently published what they purported to be a systematic review of the literature on 'isolated traumatic shaking' in infants, concluding that 'there is limited evidence that the so-called triad (encephalopathy, subdural haemorrhage, retinal haemorrhage) and therefore its components can be associated with traumatic shaking'. This flawed report, from a national body, demands a robust response. The conclusions of the original report have the potential to undermine medico-legal practice...
March 6, 2018: Archives of Disease in Childhood
Thanongsak Chaiyaso, Wimada Srisuwan, Charin Techapun, Masanori Watanabe, Shinji Takenaka
The new amylolytic oleaginous red yeast, Sporidiobolus pararoseus KX709872, produced both α-amylase (540 ± 0.09 mU/mL) and amyloglucosidase (23 ± 0.00 mU/mL) and showed good ability to directly convert rice residue from canteen waste to biomass and lipids. Effects of medium composition and cultivation conditions on growth and lipid accumulation for strain KX709872 were investigated under shaking-flask and up-scaling levels. At C:N ratio of 25:1, pH 5.45, 22.36°C, and 199.40 rpm for 7 days, volumetric production of biomass and lipids, lipid content, and lipid productivity reached 17...
March 6, 2018: Preparative Biochemistry & Biotechnology
Takuya Sugawara, Mariko Chinzei, Setsuko Numano, Chifumi Kitazaki, Munehiko Asayama
OBJECTIVE: A novel filamentous cyanobacterium, a photosynthesizing microorganism, was isolated from a river, and its unique features of flocculation and pentadecane production were characterized. RESULTS: Microscopic observations and a phylogenetic analysis with 16S rDNA revealed that this strain was a Limnothrix species denoted as the SK1-2-1 strain. Auto cell-flocculation was observed when this strain was exposed to a two-step incubation involving a standing cultivation following a shaking preincubation...
March 5, 2018: Biotechnology Letters
Sean M McClure, Patrick L Ahl, Jeffrey T Blue
PURPOSE: The purpose was to evaluate DSF for high throughput screening of protein thermal stability (unfolding/ aggregation) across a wide range of formulations. Particular focus was exploring PROTEOSTAT® - a commercially available fluorescent rotor dye - for detection of aggregation in surfactant containing formulations. Commonly used hydrophobic dyes (e.g. SYPRO™ Orange) interact with surfactants, complicating DSF measurements. METHODS: CRM197 formulations were prepared and analyzed in standard 96-well plate rT-PCR system, using SYPRO™ Orange and PROTEOSTAT® dyes...
March 5, 2018: Pharmaceutical Research
Eduardo H Gilglioni, Jung-Chin Chang, Suzanne Duijst, Simei Go, Aziza A A Adam, Ruurdtje Hoekstra, Arthur J Verhoeven, Emy L Ishii-Iwamoto, Ronald P J Oude Elferink
Primary hepatocyte culture is an important in vitro system for the study of liver functions. In vivo , hepatocytes have high oxidative metabolism. However, oxygen supply by means of diffusion in in vitro static cultures is much less than that by blood circulation in vivo . Therefore, we investigated whether hypoxia contributes to dedifferentiation and deregulated metabolism in cultured hepatocytes. To this end, murine hepatocytes were cultured under static or shaken (60 revolutions per minute) conditions in a collagen sandwich...
March 2018: Hepatology Communications
Anna Kowalska, Tomasz Boruta, Marcin Bizukojć
The application of microparticle-enhanced cultivation (MPEC) is an attractive method to control mycelial morphology, and thus enhance the production of metabolites and enzymes in the submerged cultivations of filamentous fungi. Unfortunately, most literature data deals with the spore-agglomerating species like aspergilli. Therefore, the detailed quantitative study of the morphological evolution of four different fungal species (Aspergillus terreus, Penicillium rubens, Chaetomium globosum, and Mucor racemosus) based on the digital analysis of microscopic images was presented in this paper...
March 5, 2018: MicrobiologyOpen
Prachaya Srivanitchapoom, Yuvadee Pitakpatapee, Arpakorn Suengtaworn
Since James Parkinson published his remarkable clinical observations in the "An Essay On The Shaking Palsy" in 1817, the number of diseases included in the spectrum of parkinsonian syndromes (a group of diseases that have some part of their clinical features resembling those seen in Parkinson's disease), are growing. Careful history taking, comprehensive neurological examination, and utilization of proper investigations will lead the physicians to make an accurate diagnosis of the specific disease entity present...
March 2018: Neurology India
Stefania Moretti, Alberto Greco
Studies from the embodiment perspective on language processing have shown facilitation or interference effects depending on the compatibility between verbal contents, concrete or abstract, and the motion of various parts of the body. The aim of the present study was to test whether such compatibility effects can be found when a higher cognitive process like truth evaluation is accomplished with head movements. Since nodding is a vertical head gesture typically performed with positive and affirmative responses, and shaking is a horizontal head gesture associated with negative and dissenting contents, faster response times can be expected when true information is evaluated by making a vertical head movement and false information by making a horizontal head movement...
March 1, 2018: Acta Psychologica
Joshua S Elmore, Ann M Decker, Agnieszka Sulima, Kenner C Rice, John S Partilla, Bruce E Blough, Michael H Baumann
2,5-Dimethoxyphenethylamines (2C compounds) are 5-HT2A/2C receptor agonists that induce hallucinogenic effects. N-methoxybenzylation of 2C compounds markedly increases their affinity for 5-HT2A receptors, and two such analogs, 2-(4-chloro-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25C-NBOMe) and 2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine (25I-NBOMe), have emerged in recreational drug markets. Here, we investigated the neuropharmacology of 25C-NBOMe and 25I-NBOMe in rats, as compared to their 2C analogs and the prototypical 5-HT2A/2C agonist 1-(4-iodo-2,5-dimethoxyphenyl)propan-2-amine (DOI)...
March 1, 2018: Neuropharmacology
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