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Lambeth Luke S - - 2013
Since the first application of RNA interference (RNAi) in mammalian cells, the expression of short hairpin RNAs (shRNAs) for targeted gene silencing has become a benchmark technology. Using plasmid and viral vectoring systems, the transcription of shRNA precursors that are effectively processed by the RNAi pathway can lead to potent ...
Maniar Kruti P KP Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD 21231, - - 2013
The majority of vulvar intraepithelial neoplasia (VIN) is high-grade and is related to high-risk human papillomavirus (HRHPV) (most commonly HPV 16). It is considered to be the precursor of HRHPV-related vulvar squamous cell carcinoma. Vulvar condyloma acuminatum is low-risk HPV (LRHPV)-related (most commonly types 6 and 11) and has virtually ...
Bramsen Jesper B - - 2013
Synthetic small interfering RNAs (siRNAs) have revolutionized functional genomics in mammalian cell cultures due to their reliability, efficiency, and ease of use. This success, however, has not fully translated into siRNA applications in vivo and in siRNA therapeutics where initial optimism has been dampened by a lack of efficient delivery ...
Tomer Raju - - 2013
Light sheet-based fluorescence microscopy (LSFM) is emerging as a powerful imaging technique for the life sciences. LSFM provides an exceptionally high imaging speed, high signal-to-noise ratio, low level of photo-bleaching, and good optical penetration depth. This unique combination of capabilities makes light sheet-based microscopes highly suitable for live imaging applications.Here, ...
McLachlan Sandra M SM Thyroid Autoimmune Disease Unit, Cedars-Sinai Medical Center and UCLA School of Medicine, Los Angeles, California 90048, USA. - - 2013
Thyrotropin receptor (TSHR) antibodies that stimulate the thyroid (TSAb) cause Graves' hyperthyroidism and TSHR antibodies which block thyrotropin action (TBAb) are occasionally responsible for hypothyroidism. Unusual patients switch from TSAb to TBAb (or vice versa) with concomitant thyroid function changes. We have examined case reports to obtain insight into the ...
Sætrom Pål - - 2013
Short interfering RNAs (siRNAs) have become valued tools for knocking down specific genes. As such, siRNAs are routinely used to study gene function and are also being explored as therapeutic agents. Traditionally, siRNAs are designed to target one specific gene, but this chapter describes a procedure for designing dual-targeting siRNAs ...
Trotman Winifred E - - 2013
Confocal laser scanning microscopy is commonly used to visualize and quantify protein expression. Visualization of the expression of multiple proteins in the same region via multifluorescence allows for the analysis of differential protein expression. The defining step of multifluorescence labeling is the selection of primary antibodies from different host species. ...
Liu Ying Poi - - 2013
RNA interference (RNAi) has been widely used as a tool for gene knockdown in fundamental research and for the development of new RNA-based therapeutics. The RNAi pathway is typically induced by expression of ∼22 base pair (bp) small interfering RNAs (siRNAs), which can be transfected into cells. For long-term gene ...
Takasaki Shigeru - - 2013
Short interfering RNA (siRNA) has been widely used for studying gene function in mammalian cells but varies markedly in its gene silencing efficacy. Although many design rules/guidelines for effective siRNAs based on various criteria have been reported recently, there are only a few consistencies among them. This makes it difficult ...
Evensen Lasse - - 2013
Automated multicolor fluorescence microscopy facilitates high-throughput quantitation of cellular parameters of complex, organotypic systems. In vitro co-cultured vascular cells form capillary-like networks that model facets of angiogenesis, making it an attractive alternative for anti-angiogenic drug discovery. We have adapted this angiogenesis assay system to a high-throughput format to enable automated ...
Matveeva Olga - - 2013
The design of small gene silencing RNAs with a high probability of being efficient still has some elements of an art, especially when the lowest concentration of small molecules needs to be utilized. The design of highly target-specific small interfering RNAs or short hairpin RNAs is even a greater challenging ...
Vonturkovich Michele A - - 2013
Nerve Morphometry is one tool employed in the clinical assessment of peripheral sural nerve pathological abnormalities. A new method is presented in this chapter incorporating an unbiased approach to quantitative sural nerve evaluation. Using conventional epoxy embedded nerves processed for electron microscopy, confocal microscopy, and interactive digital assessment, this method ...
Suzuki Michitaka - - 2013
Cells store excess lipid as esters in the form of triglycerides and cholesterol esters. Most lipid esters are compartmentalized in globular structures called lipid droplets. Here we describe several methods of detecting lipid droplets by fluorescence microscopy. Lipid droplets can be visualized either by staining the lipid ester core using ...
Mühlfeld Christian - - 2013
This chapter deals with the stereological quantification of structural characteristics of the lung. The aim of design-based stereological methods is the unbiased and efficient estimation of structural features without making any assumptions on the underlying nature of the biological sample. The methods are based on rigorous sampling of location and ...
Prins Mayumi L ML Department of Neurosurgery, University of California, School of Medicine, Los Angeles, CA 90095, USA. - - 2013
Among the 3.5 million annual new head injury cases is a subpopulation of children and young adults who experience repeated traumatic brain injury (TBI). The duration of vulnerability after a single TBI remains unknown, and biomarkers have yet to be determined. Decreases in glucose metabolism (cerebral metabolic rate of glucose ...
Liu Li-Ting LT School of Occupational Therapy, National Taiwan University, 100 Taipei, - - 2013
The purpose of this study is to examine the psychometric properties of the Chinese version of the Paediatric Volitional Questionnaire (PVQ-C) for use amongst preschoolers in Taiwan. Forty preschoolers with developmental delays were randomly selected from northern Taiwan, along with another 40 typically developing preschoolers. The data was analysed using ...
Shankaran Sunita S - - 2013
Real-time imaging coupled with a permeabilized cell system presents a very versatile platform to visualize the dynamic and intricate nature of nuclear envelope breakdown, one of the major morphological changes of mitosis. Here, we describe such a strategy in which the plasma membrane of cells expressing fluorescently tagged nucleoporin POM121 ...
Meisslitzer-Ruppitsch Claudia - - 2013
Correlative microscopic approaches combine the advantages of both light and electron microscopy. Here we show a correlative approach that uses the photooxidation capacity of fluorescent dyes. Through illumination with high energetic light, the chromogen diaminobenzidine is oxidized and stable deposits are formed at the sites of the former fluorescent signals, ...
Farne Hugo A HA Queen Elizabeth Hospital, University of Bristol, Bristol, UK. - - 2013
To assess the ability of C-reactive (CRP) protein, against the other commonly used metrics, to predict metronidazole treatment failure in Clostridium difficile infection. We retrospectively reviewed the case notes of 65 patients with C. difficile infection initially treated with metronidazole. Patients were grouped on the basis of outcome: those who ...
Fahim Muhammad - - 2013
RNA-mediated virus resistance is increasingly becoming a method of choice for antiviral defense in plants when effective natural resistance is unavailable. In this chapter we discuss the design principles of artificial micro RNA (amiRNA), in which a natural miRNA precursor gene is modified to target a different species of RNA, ...
Vaish Narendra - - 2013
The efficient and specific silencing of genes via RNA interference (RNAi) for functional genomics and therapeutics depends on careful consideration of the factors that affect the functionality of small interfering RNA (siRNA). These factors include (1) the length of sequence available for siRNA targeting of an mRNA, (2) the structural ...
Adler Jeremy - - 2013
The measurement of colocalization requires images of two fluorophores that are aligned, with no cross talk, and that the intensities remain within the response range of the microscope. Quantitation depends upon differentiating between the presence and absence of fluorescence, and measurements should be made within biologically relevant regions of interest. ...
Bonekamp Nina A - - 2013
Peroxisomes and mitochondria are essential subcellular organelles in mammals. Interestingly, recent studies have elucidated that these highly dynamic and plastic organelles exhibit a much closer interrelationship than previously assumed. Peroxisomes and mitochondria are metabolically linked organelles, which are cooperating and cross-talking. They share key components of their division machinery and ...
McGregor J E - - 2013
Environmental scanning electron microscopy (ESEM) (1) is an imaging technique which allows hydrated, insulating samples to be imaged under an electron beam. The resolution afforded by this technique is higher than conventional optical microscopy but lower than conventional scanning electron microscopy (CSEM). The major advantage of the technique is the ...
Mackay Joanna L - - 2013
Atomic force microscopy (AFM) is a powerful and versatile tool for probing the mechanical properties of biological samples. This chapter describes the procedures for using AFM indentation to measure the elastic moduli of living cells. We include step-by-step instructions for cantilever calibration and data acquisition using a combined AFM/optical microscope ...
Golubeva Yelena - - 2013
Laser capture microdissection (LCM) allows the precise procurement of enriched cell populations from a heterogeneous tissue, or live cell culture, under direct microscopic visualization. Histologically enriched cell populations can be procured by harvesting cells of interest directly or isolating specific cells by ablating unwanted cells. The basic components of laser ...
Falgarone Géraldine - - 2013
The role of stress in the pathophysiology of Graves' disease is suggested by several clinical observations, by recent advances in immunology and by better understanding of autoimmune diseases which provides new insights into potential effects of stress hormones on T helper cell imbalance involved in the pathogenesis of autoimmune diseases. ...
Lewis Kaitlyn N KN 1 Department of Cellular and Structural Biology, University of Texas Health Science Center at San Antonio , San Antonio, - - 2013
The oxidative stress theory of aging has been the most widely accepted theory of aging providing insights into why we age and die for over 50 years, despite mounting evidence from a multitude of species indicating that there is no direct relationship between reactive oxygen species (ROS) and longevity. Here ...
Keren Ron R Division of General Pediatrics, Center for Pediatric Clinical Effectiveness, Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA. - - 2012
To use information about prevalence, cost, and variation in resource utilization to prioritize comparative effectiveness research topics in hospital pediatrics. Retrospective analysis of administrative and billing data for hospital encounters. Thirty-eight freestanding US children's hospitals from January 1, 2004, through December 31, 2009. Children hospitalized with conditions that accounted for ...
Chakrabarti R - - 2012
Abstract Larval rohu Labeo rohita were fed four different diets: three of the diets contained Achyranthes aspera (prickly chaff-flower) seeds at 0.10% (D1), 0.25% (D2), or 0.50% (D3); the fourth diet was a control diet (D4; no A. aspera supplementation). After 70 d, the rohu were injected intraperitoneally with live ...
Horowitz Lisa M - - 2012
To develop a brief screening instrument to assess the risk for suicide in pediatric emergency department patients. A prospective, cross-sectional instrument-development study evaluated 17 candidate screening questions assessing suicide risk in young patients. The Suicidal Ideation Questionnaire served as the criterion standard. Three urban, pediatric emergency departments associated with tertiary ...
Thomas Laura A - - 2012
CONTEXT Youth with bipolar disorder (BD) and those with severe, nonepisodic irritability (severe mood dysregulation [SMD]) exhibit amygdala dysfunction during facial emotion processing. However, studies have not compared such patients with each other and with comparison individuals in neural responsiveness to subtle changes in facial emotion; the ability to process ...
Karnon Jonathan - - 2012
Regular blood transfusions for beta-thalassaemia patients lead to the accumulation of iron deposits in the body. In order to remove such deposits, iron chelation therapy is required. Subcutaneously administered deferoxamine has been the gold standard chelation therapy for over 40 years. Deferasirox is a newer chelation therapy that is taken orally ...
Puel Anne A Laboratory of Human Genetics of Infectious Diseases, Necker Branch, Inserm U980 and University Paris Descartes, Necker Medical School, Paris Sorbonne Cité, Paris, - - 2012
Chronic mucocutaneous candidiasis (CMC) is characterized by recurrent or persistent symptomatic infection of the nails, skin and mucosae mostly by Candida albicans. CMC is common in patients with profound primary T-cell immunodeficiency, who often display multiple infectious and autoimmune diseases. Patients with syndromic CMC, including autosomal dominant hyper IgE syndrome ...
Horvitz-Lennon Marcela M RAND Corporation, 4570 Fifth Ave., Suite 600, Pittsburgh, PA 15213, USA. - - 2012
This study evaluated the effect of race-ethnicity and geography on the adoption of a pharmacological innovation (long-acting injectable risperidone [LAIR]) among Medicaid beneficiaries with schizophrenia as well as the contribution of geographic location to observed racial-ethnic disparities. The data source was a claims data set from the Florida Medicaid program ...
Neudecker Ryen A - - 2012
Abstract Spring creeks are important spawning and rearing areas for wild trout, but the stable flows, cool temperatures, and high nutrient levels that characterize these unique habitats may also make them highly susceptible to establishment and proliferation of the whirling disease pathogen Myxobolus cerebralis. We evaluated the spatial and temporal ...
Green Michael F - - 2012
CONTEXT Schizophrenia remains a highly disabling disorder, but the specific determinants and pathways that lead to functional impairment are not well understood. It is not known whether these key determinants of outcome lie on 1 or multiple pathways. OBJECTIVE To evaluate theoretically based models of pathways to functional outcome starting ...
Varnado Cornelius L CL Department of Biochemistry & Cell Biology, Rice University, Houston, TX 77005, - - 2013
The worldwide blood shortage has generated a significant demand for alternatives to whole blood and packed red blood cells for use in transfusion therapy. One such alternative involves the use of acellular recombinant hemoglobin (Hb) as an oxygen carrier. Large amounts of recombinant human Hb can be expressed and purified ...
Kass David A - - 2013
Heart disease is a major cause of death and morbidity worldwide, particularly so in industrialized nations. A major contributor to its varied pathophysiology is abnormal oxidative/reductive and nitrosative stress. While oxidant-scavenging therapies for cardiovascular disease have been disappointing to date, new insights into the precise sources of oxidative/nitrosative stress, its ...
Tomás Ana M AM IBMC-Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal. - - 2013
In the single mitochondrion of protozoan trypanosomatid parasites there are several sites for the generation and elimination of reactive oxygen species (ROS), a class of molecules that exhibit a dual role in cells, either as regulatory mediators or as cytotoxic effectors. Formation of ROS in trypanosomatid mitochondria can be induced ...
Kluge Lars - - 2012
The proton affinities of hydrogen and oxygen are very similar. Therefore, it has been discussed that the proton transfer from the omnipresent H(3)(+) to molecular oxygen in the near thermoneutral reaction H(3)(+) + O(2) <--> O(2)H(+) + H(2) effectively binds the interstellar oxygen in O(2)H(+). In this work, the proton ...
Larsson Mats - - 2012
The dissociative recombination of H(3)(+) has been an intriguing problem for more than half a century. The early experiments on H(3)(+) during the first 20 years were carried out without mass analysis in decaying plasma afterglows, and thus the measured rates pertained to an uncontrolled mixture of H(3)(+) and impurity ...
Gerin M - - 2012
The Herschel-guaranteed time key programme PRobing InterStellar Molecules with Absorption line Studies (PRISMAS)(1) is providing a survey of the interstellar hydrides containing the elements C, O, N, F and Cl. As the building blocks of interstellar molecules, hydrides provide key information on their formation pathways. They can also be used ...
Indriolo Nick - - 2012
Owing to its simple chemistry, H(3)(+) is widely regarded as the most reliable tracer of the cosmic-ray ionization rate in diffuse interstellar clouds. At present, H(3)(+) observations have been made in over 50 sight lines that probe the diffuse interstellar medium (ISM) throughout the Galaxy. This small survey presents the ...
Crabtree Kyle N - - 2012
In diffuse molecular clouds, the nuclear spin temperature of H(3)(+) (approx. 30 K) is much lower than the cloud kinetic temperature (approx. 70 K). To understand this temperature discrepancy, we have measured the ratio of the hop to exchange pathways (α) in the H(3)(+) + H(2) --> H(2) + H(3)(+) reaction (which ...
Polyansky Oleg L - - 2012
The molecular ion H(3)(+) is the simplest polyatomic and poly-electronic molecular system, and its spectrum constitutes an important benchmark for which precise answers can be obtained ab initio from the equations of quantum mechanics. Significant progress in the computation of the ro-vibrational spectrum of H(3)(+) is discussed. A new, global ...
Adamowicz Ludwik - - 2012
The most accurate electronic structure calculations are performed using wave function expansions in terms of basis functions explicitly dependent on the inter-electron distances. In our recent work, we use such basis functions to calculate a highly accurate potential energy surface (PES) for the H(3)(+) ion. The functions are explicitly correlated ...
Berg Max - - 2012
We report on the recent observation of new spectral lines of cold H(3)(+) ions lying well in the visible spectral region. Transitions from the two lowest ro-vibrational levels to final levels up to 16,700 cm(-1), almost half way to the dissociation limit, have been measured, involving up to eight vibrational quanta. ...
Plasil Radek - - 2012
Formation of H(3)(+) via association of H(+) with H(2) has been studied at low temperatures using a 22-pole radiofrequency trap. Operating at hydrogen number densities from 10(11) to 10(14) cm(-3), the contributions of radiative, k(r), and ternary, k(3), association have been extracted from the measured apparent binary rate coefficients, k*=k(r)+k(3)[H(2)]. Surprisingly, ...
Stallard Tom S - - 2012
Since its discovery at Jupiter in 1988, emission from H(3)(+) has been used as a valuable diagnostic tool in our understanding of the upper atmospheres of the giant planets. One of the lasting questions we have about the giant planets is why the measured upper atmosphere temperatures are always consistently ...
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