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Jessica Cohen, Katherine Lofgren, Margaret McConnell
No abstract text is available yet for this article.
May 2017: American Economic Review
Thomas H Segall-Shapiro, Eduardo D Sontag, Christopher A Voigt
The internal environment of growing cells is variable and dynamic, making it difficult to introduce reliable parts, such as promoters, for genetic engineering. Here, we applied control-theoretic ideas to design promoters that maintained constant levels of expression at any copy number. Theory predicts that independence to copy number can be achieved by using an incoherent feedforward loop (iFFL) if the negative regulation is perfectly non-cooperative. We engineered iFFLs into Escherichia coli promoters using transcription-activator-like effectors (TALEs)...
March 19, 2018: Nature Biotechnology
Li Xie, Liqin Chen, Pan Gu, Lanlan Wei, Xuejun Kang
The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and related diseases, but their precise measurement is still a challenge. Many protocols have been described for neurotransmitter measurement by a variety of instruments, including high-pressure liquid chromatography (HPLC). However, there are shortcomings, such as complicated operation or hard-to-detect multiple targets, which cannot be avoided, and presently, the dominant analysis technique is still HPLC coupled with sensitive electrochemical or fluorimetric detection, due to its high sensitivity and good selectivity...
March 1, 2018: Journal of Visualized Experiments: JoVE
Vid Leko, Smitha Sripathy, Robin L Adrianse, Taylor Loe, Angela Park, Uyen Lao, Eric J Foss, Marisa S Bartolomei, Antonio Bedalov
Forward genetic screens using reporter genes inserted into the heterochromatin have been extensively used to investigate mechanisms of epigenetic control in model organisms. Technologies including short hairpin RNAs (shRNAs) and clustered regularly interspaced short palindromic repeats (CRISPR) have enabled such screens in diploid mammalian cells. Here we describe a large-scale shRNA screen for regulators of X-chromosome inactivation (XCI), using a murine cell line with firefly luciferase and hygromycin resistance genes knocked in at the C-terminus of the methyl CpG binding protein 2 (MeCP2) gene on the inactive X-chromosome (Xi)...
March 2, 2018: Journal of Visualized Experiments: JoVE
Markus Köhler, Patrick Oßwald, Dominik Krueger, Ryan Whitside
This manuscript describes a high-temperature flow reactor experiment coupled to the powerful molecular beam mass spectrometry (MBMS) technique. This flexible tool offers a detailed observation of chemical gas-phase kinetics in reacting flows under well-controlled conditions. The vast range of operating conditions available in a laminar flow reactor enables access to extraordinary combustion applications that are typically not achievable by flame experiments. These include rich conditions at high temperatures relevant for gasification processes, the peroxy chemistry governing the low temperature oxidation regime or investigations of complex technical fuels...
February 19, 2018: Journal of Visualized Experiments: JoVE
Aurélie Fossey-Jouenne, Carine Vergne-Vaxelaire, Anne Zaparucha
Amino alcohols are versatile compounds with a wide range of applications. For instance, they have been used as chiral scaffolds in organic synthesis. Their synthesis by conventional organic chemistry often requires tedious multi-step synthesis processes, with difficult control of the stereochemical outcome. We present a protocol to enzymatically synthetize amino alcohols starting from the readily available L-lysine in 48 h. This protocol combines two chemical reactions that are very difficult to conduct by conventional organic synthesis...
February 16, 2018: Journal of Visualized Experiments: JoVE
Xing Chen, Detlef Wolf, Juliane Siebourg-Polster, Christian Czech, Ulrike Bonati, Dirk Fischer, Omar Khwaja, Martin Strahm
Progressive and irreversible muscle atrophy characterizes Spinal Muscular Atrophy (SMA) and other similar muscle disorder diseases. Objective assessment of muscle functions is an essential and important, although challenging, prerequisite for successful clinical trials. Current clinical rating scales restrain the movement abnormalities to certain predefined coarse-grained individual items. The Kinect 3-D sensor has emerged as a low-cost and portable motion sensing technology used to capture and track people's movement in many medical and research fields...
February 12, 2018: Journal of Visualized Experiments: JoVE
Paolo Giuseppe Limoli, Enzo Maria Vingolo, Celeste Limoli, Sergio Zaccaria Scalinci, Marcella Nebbioso
This study is aimed at examining whether a suprachoroidal graft of autologous cells can improve best corrected visual acuity (BCVA) and responses to microperimetry (MY) in eyes affected by dry Age-related Macular Degeneration (AMD) over time through the production and secretion of growth factors (GFs) on surrounding tissue. Patients were randomly assigned to each study group. All patients were diagnosed with dry AMD and with BCVA equal to or greater than 1 logarithm of the minimum angle of resolution (logMAR)...
February 12, 2018: Journal of Visualized Experiments: JoVE
Takuma Iguchi, Noriyo Niino, Satoshi Tamai, Ken Sakurai, Kazuhiko Mori
RT-qPCR is one of the most common methods to assess individual target miRNAs. MiRNAs levels are generally measured relative to a reference sample. This approach is appropriate for examining physiological changes in target gene expression levels. However, absolute quantification using better statistical analysis is preferable for a comprehensive assessment of gene expression levels. Absolute quantification is still not in common use. This report describes a protocol for measuring the absolute levels of plasma miRNA, using RT-qPCR with or without pre-amplification...
February 12, 2018: Journal of Visualized Experiments: JoVE
Jianhua Ren, Yadwinder S Mann, Yuntao Zhang, Michael D Browne
Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have been thought of as a type of molecular information storage. Mass spectrometry analysis has been considered the method of choice for sequencing peptoids. Peptoids can be synthesized via solid phase chemistry using a repeating two-step reaction cycle. Here we present a method to manually synthesize oligo-peptoids and to analyze the sequence of the peptoids using tandem mass spectrometry (MS/MS) techniques...
February 21, 2018: Journal of Visualized Experiments: JoVE
Jacey L Janz Vernoski, Jack R Bjorkland, Talia J Kramer, Steven T Oczak, Alexandra L Borstad
Proprioception may be the least well measured of all contributors to the neural control of movement. New precise, reliable measures of proprioception are needed for clinical diagnosis of impairment, and to measure outcomes of proprioceptive training. The purpose of this simple, non-invasive method is to temporarily knockdown upper limb proprioception in healthy adults, to an extent that would be useful in the development and testing of upper limb proprioception measures. Knockdown models have two main advantages over studying humans with impaired proprioception: participant availability and the ability to control the extent of impairment across participants...
March 3, 2018: Journal of Visualized Experiments: JoVE
Vladimir Zeldetz, Dmitry Natanel, Matthew Boyko, Alexander Zlotnik, Honore N Shiyntum, Julia Grinshpun, Dmitry Frank, Ruslan Kuts, Evgeni Brotfain, Jochanan Peiser
Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is no existing format for studying the mechanism of action, prevention, containment, and treatment of contagious depression. The purpose of this study, therefore, was to establish the first animal model of contagious depression. Healthy rats can contract depressive behaviors if exposed to depressed rats. Depression is induced in rats by subjecting them to several manipulations of chronic unpredictable stress (CUS) over 5 weeks, as described in the protocol...
February 22, 2018: Journal of Visualized Experiments: JoVE
Ju-Chun Chang, Se Jin Lee, Jae Su Kim, Chung-Hsiung Wang, Yu-Shin Nai
The transient gene expression system is one of the most important technologies for performing protein functional analysis in the baculovirus in vitro cell culture system. This system was developed to express foreign genes under the control of the baculoviral promoter in transient expression plasmids. Furthermore, this system can be applied to a functional assay of either the baculovirus itself or foreign proteins. The most widely and commercially available transient gene expression system is developed based on the immediate-early gene (IE) promoter of Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV)...
February 22, 2018: Journal of Visualized Experiments: JoVE
Harmen B Gudde, Debra Griffiths, Kenny R Coventry
The memory game paradigm is a behavioral procedure to explore the relationship between language, spatial memory, and object knowledge. Using two different versions of the paradigm, spatial language use and memory for object location are tested under different, experimentally manipulated conditions. This allows us to tease apart proposed models explaining the influence of object knowledge on spatial language (e.g., spatial demonstratives), and spatial memory, as well as understanding the parameters that affect demonstrative choice and spatial memory more broadly...
February 19, 2018: Journal of Visualized Experiments: JoVE
Xinyi Ding, Yuxiang Sun, Yanming Chen, Wanchuan Ding, Steven Emory, Tianhao Li, Zixing Xu, Ning Han, Jun Wang, Gang Ruan
Micellar nanocrystals (micelles with encapsulated nanocrystals) have become an emerging major class of nanobiomaterials. We describe a method of fabricating micellar nanocrystals based on combining top-down electrospray, bottom-up self-assembly, and solvent-based structure control. This method involves first using electrospray to generate uniform ultrafine liquid droplets, each of which functions as a micro-reactor in which self-assembly reaction occurs forming micellar nanocrystals, with the structures (micelle shape and nanocrystal encapsulation) controlled by the organic solvent used...
February 11, 2018: Journal of Visualized Experiments: JoVE
Andrey Zavadsky, Lev Shemer
This manuscript describes an experimental procedure that allows obtaining diverse quantitative information on temporal and spatial evolution of water waves excited by time-dependent and steady wind forcing. Capacitance-type wave gauge and Laser Slope Gauge (LSG) are used to measure instantaneous water surface elevation and two components of the instantaneous surface slope at a number of locations along the test section of a wind-wave facility. The computer-controlled blower provides airflow over the water in the tank whose rate can vary in time...
February 13, 2018: Journal of Visualized Experiments: JoVE
Mireille Laforge, Ricardo Silvestre, Vasco Rodrigues, Julie Garibal, Laure Campillo-Gimenez, Shahul Mouhamad, Valérie Monceaux, Marie-Christine Cumont, Henintsoa Rabezanahary, Alain Pruvost, Anabela Cordeiro-da-Silva, Bruno Hurtrel, Guido Silvestri, Anna Senik, Jérôme Estaquier
Apoptosis has been proposed as a key mechanism responsible for CD4+ T cell depletion and immune dysfunction during HIV infection. We demonstrated that Q-VD-OPH, a caspase inhibitor, inhibits spontaneous and activation-induced death of T cells from SIV-infected rhesus macaques (RMs). When administered during the acute phase of infection, Q-VD-OPH was associated with (a) reduced levels of T cell death, (b) preservation of CD4+/CD8+ T cell ratio in lymphoid organs and in the gut, (c) maintenance of memory CD4+ T cells, and (d) increased specific CD4+ T cell response associated with the expression of cytotoxic molecules...
March 19, 2018: Journal of Clinical Investigation
Cicely Marston, Alicia Renedo, Gertrude Nsorma Nyaaba, Kazuyo Machiyama, Placide Tapsoba, John Cleland
CONTEXT: According to Demographic and Health Survey (DHS) data, highly educated urban women in some West African countries simultaneously have low rates of both contraceptive use and fertility-suggesting that the DHS may not be capturing a complete picture of women's contraceptive practices. METHODS: Individual in-depth interviews and focus group discussions were conducted with a total of 48 women aged 18-49 in Accra, Ghana, who had at least a secondary education to explore their reproductive lives and relationships, and their views on and use of fertility regulation strategies...
September 1, 2017: International Perspectives on Sexual and Reproductive Health
Mustafa Erdoğan Cicik, Cezmi Doğan, Selim Bölükbaşı, Mehmet Necdet Cinhüseyinoğlu, Osman Şevki Arslan
BACKGROUND: Pediatric cataract surgery differs substantially from adult cataract surgery. Numerous studies have focused on reducing the development of postoperative complications. AIMS: To compare two intraocular lens implantation techniques used in pediatric cataract surgery in terms of postoperative complications. STUDY DESIGN: Case-control study. METHODS: Patients who underwent pediatric cataract surgery and intraocular lens implantation between 2008 and 2016 were evaluated in this retrospective study...
March 15, 2018: Balkan Medical Journal
Mustafa Erdoğan Cicik, Cezmi Doğan, Osman Şevki Arslan
BACKGROUND: Development of endophthalmitis during cataract surgery is one of the most severe complications and it may also result in blindness and globe loss. AIMS: To evaluate the effect of an ophthalmic cefuroxime solution (Aprokam® ) that is prophylactically used in phacoemulsification surgery performed due to cataract that eventually occurs in patients with or without penetrating keratoplasty. STUDY DESIGN: Case-control study. METHODS: In this retrospective study, patients who had undergone penetrating keratoplasty and for whom phacoemulsification surgery was performed due to cataract and an intracameral ophthalmic cefuroxime solution (Aprokam® ) was administered for prophylaxis were evaluated in comparison with patients with cataract without keratoplasty...
March 15, 2018: Balkan Medical Journal
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