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We show that the resonance shifts in the fluorescence of a cold gas of rubidium atoms substantially differ from those of thermal atomic ensembles that obey the standard continuous medium electrodynamics. The analysis is based on large-scale microscopic numerical simulations and experimental measurements of the resonance shifts in a steady-state response in light propagation.
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We measure the coherent scattering of light by a cloud of laser-cooled atoms with a size comparable to the wavelength of light. By interfering a laser beam tuned near an atomic resonance with the field scattered by the atoms, we observe a resonance with a redshift, a broadening, and a saturation of the extinction for increasing atom numbers. We attribute these features to enhanced light-induced dipole-dipole interactions in a cold, dense atomic ensemble that result in a failure of standard predictions such as the "cooperative Lamb shift". The description of the atomic cloud by a mean-field model based on the Lorentz-Lorenz formula that ignores scattering events where light is scattered recurrently by the same atom and by a microscopic discrete dipole model that incorporates these effects lead to progressively closer agreement with the observations, despite remaining differences.
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We study the emergence of collective scattering in the presence of dipole-dipole interactions when we illuminate a cold cloud of rubidium atoms with a near-resonant and weak intensity laser. The size of the atomic sample is comparable to the wavelength of light. When we gradually increase the number of atoms from 1 to ~450, we observe a broadening of the line, a small redshift and, consistently with these, a strong suppression of the scattered light with respect to the noninteracting atom case. We compare our data to numerical simulations of the optical response, which include the internal level structure of the atoms.
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We have implemented the Gedanken experiment of an individual atom scattering a wave packet of near-resonant light, and measured the associated Wigner time delay as a function of the frequency of the light. In our apparatus, the atom behaves as a two-level system and we have found delays as large as 42 ns at resonance, limited by the lifetime of the excited state. This delay is an important parameter in the problem of collective near-resonant scattering by an ensemble of interacting particles, which is encountered in many areas of physics.
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Over the past decade, Taxol and its closely related structural analogue Taxotere have emerged as very important antitumor agents. Their widespread use in the treatment of a variety of cancer types, their likely approval for the treatment of additional forms of cancer, and their use at earlier stages of intervention will lead to increased demand for these drugs in the future. Because of yield considerations, Taxol and Taxotere are currently derived via semisynthesis from the advanced taxoid 10-deacetylbaccatin III, which must be isolated from yew (Taxus) trees. Thus, efforts are underway to produce Taxol (and other advanced taxoids for use in semisynthesis) by alternate, biotechnological means. This article provides a current overview of research on taxoid biosynthesis and an assessment of bioengineering applications for taxoid production in yew cell culture.
Assuntos
Biotecnologia , Paclitaxel/biossíntese , Estrutura Molecular , Paclitaxel/química , Paclitaxel/metabolismoRESUMO
In order to clarify the role of the purine 2-amino group in the recognition of DNA by small molecules we have examined the binding of actinomycin D and echinomycin to artificial DNA molecules asymmetrically substituted with inosine and/or 2,6-diaminopurine (DAP) in one of the complementary strands. These DNAs, prepared by a method based upon PCR, present various potential sites for antibiotic binding, including several containing only a single purine 2-amino group in different configurations. The results show unambiguously that the presence of two 2-amino groups is mandatory for binding of actinomycin D to double-stranded DNA. In the case of echinomycin only one purine 2-amino group is required for remarkably strong binding to the asymmetric TpDAP.TpA dinucleotide step, but the CpDAP.TpI step (which also contains only a single purine-2 amino group) does not afford a binding site. Evidently, removing a 2-amino group (G-->I substitution) is dominant over adding one (A-->DAP substitution). No sequences containing just a single guanine residue are acceptable. The possibility is raised that replacing guanosine with inosine may do more than remove a group endowed with hydrogen bonding capability and interfere with ligand binding in other ways. The new methodology developed to construct asymmetrically substituted DNA substrates for this work provides a novel strategy that should be generally applicable for studying ligand-DNA interactions, beyond the specific interest in drug binding to DNA, and may help to elucidate how proteins and oligonucleotides recognize their target sites.
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2-Aminopurina/análogos & derivados , DNA/química , Dactinomicina/química , Equinomicina/química , Inosina , Composição de Bases , Sítios de Ligação , Citosina , Pegada de DNA , Primers do DNA , Ligação de Hidrogênio , Hipoxantina , Dados de Sequência Molecular , Ácidos Nucleicos Heteroduplexes/química , Oligodesoxirribonucleotídeos/síntese química , Oligodesoxirribonucleotídeos/química , Reação em Cadeia da Polimerase , TiminaRESUMO
A central feature in the biosynthesis of Taxol is oxygenation at multiple positions of the taxane core structure, reactions that are considered to be mediated by cytochrome P450-dependent monooxygenases. A PCR-based differential display-cloning approach, using Taxus (yew) cells induced for Taxol production, yielded a family of related cytochrome P450 genes, one of which was assigned as a taxane 10 beta-hydroxylase by functional expression in yeast. The acquired clones that did not function in yeast were heterologously expressed by using the Spodoptera fugiperda-baculovirus-based system and were screened for catalytic capability by using taxa-4(20),11(12)-dien-5 alpha-ol and its acetate ester as test substrates. This approach allowed identification of one of the cytochrome P450 clones (which bore 63% deduced sequence identity to the aforementioned taxane 10 beta-hydroxylase) as a taxane 13 alpha-hydroxylase by chromatographic and spectrometric characterization of the corresponding recombinant enzyme product. The demonstration of a second relevant hydroxylase from the induced family of cytochrome P450 genes validates this strategy for elucidating the oxygenation steps of taxane diterpenoid (taxoid) metabolism. Additionally, substrate specificity studies with the available cytochrome P450 hydroxylases now indicate that there is likely more than one biosynthetic route to Taxol in yew species.