RESUMEN
A novel DSN-depletion method allows elimination of selected sequences from full-length-enriched cDNA libraries. Depleted cDNA can be applied for subsequent EST sequencing, expression cloning, and functional screening approaches. The method employs specific features of the kamchatka crab duplex-specific nuclease (DSN). This thermostable enzyme is specific for double-stranded (ds) DNA, and is thus used for selective degradation of ds DNA in complex nucleic acids. DSN depletion is performed prior to library cloning, and includes the following steps: target cDNA is mixed with excess driver DNA (representing fragments of the genes to be eliminated), denatured, and allowed to hybridize. During hybridization, driver molecules form hybrids with the target sequences, leading to their removal from the ss DNA fraction. Next, the ds DNA fraction is hydrolyzed by DSN, and the ss fraction is amplified using long-distance PCR. DSN depletion has been tested in model experiments.
Asunto(s)
Anomuros/enzimología , ADN Complementario/metabolismo , Desoxirribonucleasas/metabolismo , Biblioteca de Genes , Animales , Anomuros/genética , Anomuros/metabolismo , Antozoos/enzimología , Antozoos/genética , ADN Complementario/genética , Desoxirribonucleasas/genética , Femenino , Humanos , Hibridación de Ácido Nucleico , Hidrolasas Diéster Fosfóricas/genética , Hidrolasas Diéster Fosfóricas/metabolismo , Placenta/enzimología , Placenta/metabolismo , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido SimpleRESUMEN
Pixelwise linear prediction using backward-adaptive least-squares or weighted least-squares estimation of prediction coefficients is currently among the state-of-the-art methods for lossless image compression. While current research is focused on mean intensity prediction of the pixel to be transmitted, best compression requires occurrence probability estimates for all possible intensity values. Apart from common heuristic approaches, we show how prediction error variance estimates can be derived from the (weighted) least-squares training region and how a complete probability distribution can be built based on an autoregressive image model. The analysis of image stationarity properties further allows deriving a novel formula for weight computation in weighted least-squares proofing and generalizing ad hoc equations from the literature. For sparse intensity distributions in non-natural images, a modified image model is presented. Evaluations were done in the newly developed C++ framework volumetric, artificial, and natural image lossless coder (Vanilc), which can compress a wide range of images, including 16-bit medical 3D volumes or multichannel data. A comparison with several of the best available lossless image codecs proofs that the method can achieve very competitive compression ratios. In terms of reproducible research, the source code of Vanilc has been made public.
RESUMEN
The management of medical emergency, in particular cardiac emergency, requests prompt intervention and the possibility to communicate in real time from the emergency area / ambulance to the hospital as much diagnostic information as possible about the patient. This would enable a prompt emergency diagnosis and operation and the possibility to prepare the appropriate actions in the suitable hospital department. To address this scenario, the CONCERTO European project proposed a wireless communication system based on a novel cross-layer architecture, including the integration of building blocks for medical media content fusion, delivery and access. This paper describes the proposed system architecture, outlining the developed components and mechanisms, and the evaluation of the proposed system, carried out in a hospital with the support of medical staff. The technical results and the feedback received highlight the impact of the CONCERTO approach in the healthcare domain, in particular in enabling a prompt and reliable diagnosis in challenging medical emergency scenarios.
Asunto(s)
Redes de Comunicación de Computadores , Sistemas de Computación , Urgencias Médicas , Multimedia , Ambulancias , Hospitales , Humanos , Reproducibilidad de los Resultados , Grabación en VideoRESUMEN
A total of 53 N-benzoylated phenoxazines and phenothiazines, including their S-oxidized analogues, were synthesized and evaluated for antiproliferative activity, interaction with tubulin, and cell cycle effects. Potent inhibitors of multiple cancer cell lines emerged with the 10-(4-methoxybenzoyl)-10H-phenoxazine-3-carbonitrile (33b, IC(50) values in the range of 2-15 nM) and the isovanillic analogue 33c. Seventeen compounds strongly inhibited tubulin polymerization with activities higher than or comparable to those of the reference compounds such as colchicine. Concentration-dependent flow cytometric studies revealed that inhibition of K562 cell growth was associated with an arrest in the G2/M phases of the cell cycle, indicative of mitotic blockade. Structure-activity relationship studies showed that best potencies were obtained with agents bearing a methoxy group placed para at the terminal phenyl ring and a 3-cyano group in the phenoxazine. A series of analogues highlight not only the phenoxazine but also the phenothiazine structural scaffold as valuable pharmacophores for potent tubulin polymerization inhibitors, worthy of further investigation.