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Comprehensive mutational scanning of the p53 coding region by two- dimensional gene scanning 总被引:2,自引:0,他引:2
A comprehensive mutational scanning test for the p53 coding region based on
multiplex PCR and two-dimensional DNA electrophoresis was designed and
evaluated. In a 2-step multiplex PCR, the p53 coding region (exons 2-11)
was amplified as a single 8646-bp fragment by long- distance PCR in step
one. This fragment served as a template for the subsequent co-amplification
of the individual exons in two multiplex groups in step two. The multiplex
products were then separated, first on the basis of size in non-denaturant
polyacrylamide gels and then on the basis of sequence by denaturing
gradient gel electrophoresis (DGGE). Primers for optimal PCR, melting
behavior and 2-D gel distribution were designed using a recently developed
computer program. The resulting two-dimensional gene scanning (TDGS) test
was evaluated by screening, in a blinded fashion, 29 coded DNA samples from
Li- Fraumeni syndrome patients with previously identified germline
mutations. All mutations were correctly detected. This assay provides an
accurate, cost-effective and non-radioactive method for simultaneous
mutational scanning of all p53 coding exons.
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Tao Peng Ghislain M. C. Bonamy Estelle Glory-Afshar Daniel R. Rines Sumit K. Chanda Robert F. Murphy 《Proceedings of the National Academy of Sciences of the United States of America》2010,107(7):2944-2949
Many proteins or other biological macromolecules are localized to more than one subcellular structure. The fraction of a protein in different cellular compartments is often measured by colocalization with organelle-specific fluorescent markers, requiring availability of fluorescent probes for each compartment and acquisition of images for each in conjunction with the macromolecule of interest. Alternatively, tailored algorithms allow finding particular regions in images and quantifying the amount of fluorescence they contain. Unfortunately, this approach requires extensive hand-tuning of algorithms and is often cell type-dependent. Here we describe a machine-learning approach for estimating the amount of fluorescent signal in different subcellular compartments without hand tuning, requiring only the acquisition of separate training images of markers for each compartment. In testing on images of cells stained with mixtures of probes for different organelles, we achieved a 93% correlation between estimated and expected amounts of probes in each compartment. We also demonstrated that the method can be used to quantify drug-dependent protein translocations. The method enables automated and unbiased determination of the distributions of protein across cellular compartments, and will significantly improve imaging-based high-throughput assays and facilitate proteome-scale localization efforts. 相似文献
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Seitz DP Brisbin S Herrmann N Rapoport MJ Wilson K Gill SS Rines J Le Clair K Conn D 《Journal of the American Medical Directors Association》2012,13(6):503-506.e2
BackgroundNonpharmacological therapies are often recommended as a first-line treatment for neuropsychiatric symptoms (NPS) of dementia in long term care (LTC); however, little is known about which nonpharmacological interventions are most effective for NPS in LTC or the feasibility of interventions, given the availability of resources in typical LTC environments.MethodsWe searched the electronic databases MEDLINE, EMBASE, PsychINFO (1980–2010), the Cochrane Library, and Google Scholar using keywords and medical subject headings for randomized, controlled trials evaluating nonpharmacological interventions for NPS conducted in LTC settings. Change in severity of NPS symptoms was evaluated through the NPS outcomes measures reported in studies. We assessed study quality and described the feasibility of interventions based on various aspects of study design.ResultsA total of 40 studies met inclusion criteria. Sixteen (40%) of 40 included studies reported statistically significant results in favor of nonpharmacological interventions on at least one measure of NPS. These interventions included staff training in NPS management strategies, mental health consultation and treatment planning, exercise, recreational activities, and music therapy or other forms of sensory stimulation. Many of the studies had methodological limitations that placed them at potential risk of bias. Most interventions (n = 30, 75%) required significant resources from services outside of LTC or significant time commitments from LTC nursing staff for implementation.ConclusionsThere are several nonpharmacological interventions that may be effective for NPS in LTC, although there are a limited number of large-scale, high-quality studies in this area. The feasibility of some interventions will be limited in many LTC settings and further research into practical and sustainable interventions for NPS in LTC is required to improve usage of these important treatments. 相似文献
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This study was designed to investigate the reliability of real-ear measurements of sound pressure level (SPL) and to compare these values with two coupler measures of SPL. A commercially available probe tube microphone system was used to measure real ear SPL in both children and adults. Test-retest reliability decreased as a function of frequency for both groups and, in general, was slightly poorer for the children. For both groups, coupler to real ear differences were larger for the 2 cm3 coupler than for the reduced volume coupler; however, no significant differences were observed between groups. In addition, a measure of ear canal volume was not found to be a good predictor of coupler to real ear discrepancies. 相似文献
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Albaugh J Amargo I Capelson R Flaherty E Forest C Goldstein I Jensen PK Jones K Kloner R Lewis J Mullin S Payton T Rines B Rosen R Sadovsky R Snow K Vetrosky D;University of Medicine Dentistry of New Jersey 《Urologic nursing》2002,22(4):217-31; quiz 232
A consensus guidelines model was developed for managing erectile dysfunction (ED) by urology nurses and other health care clinicians. The model emphasizes identification of ED in all males older than 40, education and counseling as keys to intervention, and inclusion of partners whenever possible. These guidelines are intended to provide a comprehensive care model for ED patients and their partners, which would be optimally cost effective and clinically relevant. If successful, this new model will allow quality sexual health care to be provided to increasing numbers of patients and their partners. 相似文献
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