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121.
122.
Pospíšil L Hromadová M Fanelli N Valášek M Kolivoška V Gál M 《Physical chemistry chemical physics : PCCP》2011,13(10):4365-4371
A long organic molecule 1 with five bipyridinium functions separated by benzene rings (extended viologen) undergoes a reversible multi-step electron transfer. Here we show that this decacation accepts electrons at the heterogeneous interface with the occurrence of the periodically changing electric reduction currents. According to the applied bias voltage the observed current-time dependence changes from chaotic through periodic and irregular to sinusoidal and finally to monotonous. A careful choice of the controlling parameters yields the sustained periodic sinusoidal currents lasting for a prolonged time. Oscillations stem from a mutual interplay of the heterogeneous supply of electrons and the homogeneous redox reactions (disproportionation) between the transient redox forms. In difference to many other electrochemical oscillating systems the described oscillations do not require any additional external impedance. The principle of these oscillatory currents may serve as a model of a truly 'molecular oscillator'. 相似文献
123.
Zhong Q Bhattacharya S Kotsopoulos S Olson J Taly V Griffiths AD Link DR Larson JW 《Lab on a chip》2011,11(13):2167-2174
Quantitative polymerase chain reactions (qPCR) based on real-time PCR constitute a powerful and sensitive method for the analysis of nucleic acids. However, in qPCR, the ability to multiplex targets using differently colored fluorescent probes is typically limited to 4-fold by the spectral overlap of the fluorophores. Furthermore, multiplexing qPCR assays requires expensive instrumentation and most often lengthy assay development cycles. Digital PCR (dPCR), which is based on the amplification of single target DNA molecules in many separate reactions, is an attractive alternative to qPCR. Here we report a novel and easy method for multiplexing dPCR in picolitre droplets within emulsions-generated and read out in microfluidic devices-that takes advantage of both the very high numbers of reactions possible within emulsions (>10(6)) as well as the high likelihood that the amplification of only a single target DNA molecule will initiate within each droplet. By varying the concentration of different fluorogenic probes of the same color, it is possible to identify the different probes on the basis of fluorescence intensity. Adding multiple colors increases the number of possible reactions geometrically, rather than linearly as with qPCR. Accurate and precise copy numbers of up to sixteen per cell were measured using a model system. A 5-plex assay for spinal muscular atrophy was demonstrated with just two fluorophores to simultaneously measure the copy number of two genes (SMN1 and SMN2) and to genotype a single nucleotide polymorphism (c.815A>G, SMN1). Results of a pilot study with SMA patients are presented. 相似文献
124.
Quantitative and sensitive detection of rare mutations using droplet-based microfluidics 总被引:2,自引:0,他引:2
Pekin D Skhiri Y Baret JC Le Corre D Mazutis L Salem CB Millot F El Harrak A Hutchison JB Larson JW Link DR Laurent-Puig P Griffiths AD Taly V 《Lab on a chip》2011,11(13):2156-2166
Somatic mutations within tumoral DNA can be used as highly specific biomarkers to distinguish cancer cells from their normal counterparts. These DNA biomarkers are potentially useful for the diagnosis, prognosis, treatment and follow-up of patients. In order to have the required sensitivity and specificity to detect rare tumoral DNA in stool, blood, lymph and other patient samples, a simple, sensitive and quantitative procedure to measure the ratio of mutant to wild-type genes is required. However, techniques such as dual probe TaqMan(?) assays and pyrosequencing, while quantitative, cannot detect less than ~1% mutant genes in a background of non-mutated DNA from normal cells. Here we describe a procedure allowing the highly sensitive detection of mutated DNA in a quantitative manner within complex mixtures of DNA. The method is based on using a droplet-based microfluidic system to perform digital PCR in millions of picolitre droplets. Genomic DNA (gDNA) is compartmentalized in droplets at a concentration of less than one genome equivalent per droplet together with two TaqMan(?) probes, one specific for the mutant and the other for the wild-type DNA, which generate green and red fluorescent signals, respectively. After thermocycling, the ratio of mutant to wild-type genes is determined by counting the ratio of green to red droplets. We demonstrate the accurate and sensitive quantification of mutated KRAS oncogene in gDNA. The technique enabled the determination of mutant allelic specific imbalance (MASI) in several cancer cell-lines and the precise quantification of a mutated KRAS gene in the presence of a 200,000-fold excess of unmutated KRAS genes. The sensitivity is only limited by the number of droplets analyzed. Furthermore, by one-to-one fusion of drops containing gDNA with any one of seven different types of droplets, each containing a TaqMan(?) probe specific for a different KRAS mutation, or wild-type KRAS, and an optical code, it was possible to screen the six common mutations in KRAS codon 12 in parallel in a single experiment. 相似文献
125.
Dr. Valérie Alain‐Rizzo Delphine Drouin‐Kucma Cédric Rouxel Dr. Issa Samb Jérémy Bell Dr. Patrick Y. Toullec Dr. Véronique Michelet Dr. Isabelle Leray Dr. Mireille Blanchard‐Desce 《化学:亚洲杂志》2011,6(4):1080-1091
A series of rod‐shaped and related three‐branched push–pull derivatives containing phosphane oxide or phosphane sulfide (PO or PS)—as an electron‐withdrawing group conjugated to electron‐donating groups, such as amino or ether groups, with a conjugated rod consisting of arylene–vinylene or arylene–ethynylene building blocks—were prepared. These compounds were efficiently synthesized by a Grignard reaction followed by Sonogashira coupling. Their photophysical properties including absorption, emission, time‐resolved fluorescence, and two‐photon absorption (TPA) were investigated with special attention to structure–property relationships. These fluorophores show high fluorescence quantum yields and solvent‐dependent experiments reveal that efficient intramolecular charge transfer occurs upon excitation, thereby leading to highly polar excited states, the polarity of which can be significantly enhanced by playing on the end groups and conjugated linker. Rod‐shaped and related three‐branched systems show similar fluorescence properties in agreement with excitation localization on one of the push–pull branches. By using stronger electron donors or replacing the arylene–ethynylene linkers with an arylene–vinylene one induces significant redshifts of both the low‐energy one‐photon absorption and TPA bands. Interestingly, a major enhancement in TPA responses is observed, whereas OPA intensities are only weakly affected. Similarly, phosphane oxide derivatives show similar OPA responses than the corresponding sulfides but their TPA responses are significantly larger. Finally, the electronic coupling between dipolar branches promoted by common PO or PS acceptor moieties induces either slight enhancement of the TPA responses or broadening of the TPA band in the near infrared (NIR) region. Such behavior markedly contrasts with triphenylamine‐core‐mediated coupling, which gives evidence for the different types of interactions between branches. 相似文献
126.
Fleury BG Figueiredo L Marcoli MI Teixeira VL Ferreira AB Pinto AC 《Natural product communications》2011,6(5):667-672
The distribution of fatty acids in 13 species of macroalgae (Chlorophyta, Ochrophyta and Rhodophyta) and 1 seagrass (Spartina sp), collected on the Rio de Janeiro state coast was determined. The results were evaluated in search of correlations between the taxonomic and phylogenetic position of these macrophytes. Statistical analyses showed the effectiveness as taxonomic and phylogenetic markers of the distribution of the methyl fatty acid esters in these macrophytes. 相似文献
127.
128.
Recursive density-matrix perturbation theory [A.M.N. Niklasson and M. Challacombe, Phys. Rev. Lett. 92, 193001 (2004)] provides an efficient framework for the linear scaling computation of materials response properties [V. Weber, A.M.N. Niklasson, and M. Challacombe, Phys. Rev. Lett. 92, 193002 (2004)]. In this article, we generalize the density-matrix perturbation theory to include properties computed with a perturbation-dependent nonorthogonal basis. Such properties include analytic derivatives of the energy with respect to nuclear displacement, as well as magnetic response computed with a field-dependent basis. The theory is developed in the context of linear scaling purification methods, which are briefly reviewed. 相似文献
129.
Gaál V Kilár F Acs B Szijjártó Z Kocsis B Kustos I 《Colloids and surfaces. B, Biointerfaces》2005,45(3-4):125-130
Implantation of artificial intraocular lenses into the eye during ophthalmic surgical procedures ensures an unliving surface on which bacterial pathogens may attach and form biofilms. Despite antibiotic treatment bacteria growing in biofilms might cause inflammation and serious complications. In this study the adhesive ability of 7 Staphylococcus aureus and 11 coagulase-negative Staphylococcus (CNS) strains to the surface of acrylic intraocular lenses had been examined by the ultrasonic method. In untreated cases adhesion of the S. aureus and CNS strains did not differ significantly. We could not demonstrate significant differences between the adhesive ability of the standard strains and the clinical isolates. In this study a single – 60 min long – antibiotic (ciprofloxacin and tobramycin) treatment had been applied, that correlate well with the single or intermittant antibiotic prophylaxis of patients. Ciprofloxacin administration was able to reduce significantly the number of attached cells on the surface of acrylic lenses both in the case of S. aureus and CNS strains. Dependence of the effect from concentration could also be demonstrated. Tobramycin treatment was able to inhibit significantly the attachment of S. aureus cells. Despite the debate on antibiotic prophylaxis we presented in our experiments that a single antibiotic administration can decrease the attachment of bacterial cells to the surface of acrylic intraocular lenses, and might be effective in the prevention of postoperative endophthalmitis, that is a rare but serious complication of ophthalmic surgery. 相似文献
130.