Upper level and phase controlling group velocity in V-type system

With the external field coupling the two upper levels, we investigate the light pulse propagation properties with weak probe field in a V-type system. Due to the external upper level (UL) coupling field, the dispersion of the system has been influenced by the relative phase. It is shown that the UL field and the relative phase can be regarded as switches to manipulate light propagation between subluminal and superluminal. (c) 2007 Elsevier B.V. All rights reserved.

Controlling the group velocity in a five-level K-type atomic system

The properties of a five-level K-type system are investigated. With the controlling fields, the properties of the dispersion and absorption of the system are changed greatly. The system can produce anomalous dispersion regions with absorption and normal dispersion regions with absorption or transparency. Furthermore, the group velocity can be varied from subluminal to superluminal by varying the intensity of the controlling field and the probe detunings in principle. (C) 2008 Elsevier B.V. All rights reserved.

DNA-mediated oxidation of transcription factor p53

Transcription factor p53 is the most commonly altered gene in human cancer. As a redox-active protein in direct contact with DNA, p53 can directly sense oxidative stress through DNA-mediated charge transport. Electron hole transport occurs with a shallow distance dependence over long distances through the π-stacked DNA bases, leading to the oxidation and dissociation of DNA-bound p53. The extent of p53 dissociation depends upon the redox potential of the response element DNA in direct contact with each p53 monomer. The DNA sequence dependence of p53 oxidative dissociation was examined by electrophoretic mobility shift assays using radiolabeled oligonucleotides containing both synthetic and human p53 response elements with an appended anthraquinone photooxidant. Greater p53 dissociation is observed from DNA sequences containing low redox potential purine regions, particularly guanine triplets, within the p53 response element. Using denaturing polyacrylamide gel electrophoresis of irradiated anthraquinone-modified DNA, the DNA damage sites, which correspond to locations of preferred electron hole localization, were determined. The resulting DNA damage preferentially localizes to guanine doublets and triplets within the response element. Oxidative DNA damage is inhibited in the presence of p53, however, only at DNA sites within the response element, and therefore in direct contact with p53. From these data, predictions about the sensitivity of human p53-binding sites to oxidative stress, as well as possible biological implications, have been made. On the basis of our data, the guanine pattern within the purine region of each p53-binding site determines the response of p53 to DNA-mediated oxidation, yielding for some sequences the oxidative dissociation of p53 from a distance and thereby providing another potential role for DNA charge transport chemistry within the cell.

To determine whether the change in p53 response element occupancy observed in vitro also correlates in cellulo, chromatin immunoprecipition (ChIP) and quantitative PCR (qPCR) were used to directly quantify p53 binding to certain response elements in HCT116N cells. The HCT116N cells containing a wild type p53 were treated with the photooxidant [Rh(phi)2bpy]³⁺, Nutlin-3 to upregulate p53, and subsequently irradiated to induce oxidative genomic stress. To covalently tether p53 interacting with DNA, the cells were fixed with disuccinimidyl glutarate and formaldehyde. The nuclei of the harvested cells were isolated, sonicated, and immunoprecipitated using magnetic beads conjugated with a monoclonal p53 antibody. The purified immounoprecipiated DNA was then quantified via qPCR and genomic sequencing. Overall, the ChIP results were significantly varied over ten experimental trials, but one trend is observed overall: greater variation of p53 occupancy is observed in response elements from which oxidative dissociation would be expected, while significantly less change in p53 occupancy occurs for response elements from which oxidative dissociation would not be anticipated.

The chemical oxidation of transcription factor p53 via DNA CT was also investigated with respect to the protein at the amino acid level. Transcription factor p53 plays a critical role in the cellular response to stress stimuli, which may be modulated through the redox modulation of conserved cysteine residues within the DNA-binding domain. Residues within p53 that enable oxidative dissociation are herein investigated. Of the 8 mutants studied by electrophoretic mobility shift assay (EMSA), only the C275S mutation significantly decreased the protein affinity (KD) for the Gadd45 response element. EMSA assays of p53 oxidative dissociation promoted by photoexcitation of anthraquinone-tethered Gadd45 oligonucleotides were used to determine the influence of p53 mutations on oxidative dissociation; mutation to C275S severely attenuates oxidative dissociation while C277S substantially attenuates dissociation. Differential thiol labeling was used to determine the oxidation states of cysteine residues within p53 after DNA-mediated oxidation. Reduced cysteines were iodoacetamide labeled, while oxidized cysteines participating in disulfide bonds were ¹³C₂D₂-iodoacetamide labeled. Intensities of respective iodoacetamide-modified peptide fragments were analyzed using a QTRAP 6500 LC-MS/MS system, quantified with Skyline, and directly compared. A distinct shift in peptide labeling toward ¹³C₂D₂-iodoacetamide labeled cysteines is observed in oxidized samples as compared to the respective controls. All of the observable cysteine residues trend toward the heavy label under conditions of DNA CT, indicating the formation of multiple disulfide bonds potentially among the C124, C135, C141, C182, C275, and C277. Based on these data it is proposed that disulfide formation involving C275 is critical for inducing oxidative dissociation of p53 from DNA.

Tecnologias não-invasivas de cuidado de enfermagem obstétrica no suporte físico à parturiente: critérios e efeitos esperados

Este estudo buscou identificar os critérios utilizados pelas enfermeiras obstétricas para empregar as tecnologias não-invasivas de cuidado no suporte físico à parturiente e quais são seus efeitos esperados. Para tanto, se realizou uma pesquisa quantitativa exploratória do tipo survey, que abordou as práticas/cuidados fornecidas pelas enfermeiras obstétrica durante a assistência ás parturientes que tinham relação com o suporte físico. Estas foram agrupadas em: suporte relacionado ao ambiente; suporte ao posicionamento (livre movimentação e adoção de posturas verticais, deambulação, movimentos pélvicos, posição de cócoras, e posição de quatro apoios); suporte aos estímulos táteis (massagens, compressas frias/mornas, banho morno de aspersão e imersão); e suporte energético (oferta de alimento). Para tanto, utilizou-se um questionário, que foi disponibilizado via internet, após a criação de um domínio e web site próprio para tal finalidade. A população-alvo constituiu-se de enfermeiras obstétricas que atuam no cuidado da parturiente em território nacional, sendo que estas foram convidadas a participar, entre os meses de julho e setembro de 2009, através de e-mails survey individuais ou coletivos. Participaram do estudo120 profissionais, sendo que 45,8% foram excluídos automaticamente pelo sistema, pois não possuíam os critérios de inclusão. Dos participantes elegíveis (65), 33,8% responderam somente o teste de elegibilidade, 4,6% responderam parcialmente e 61,6% responderam completamente o questionário. Os resultados demonstraram que alguns aspectos relacionados ao conceito ainda encontram-se pouco compreendidos pelas profissionais da área, entretanto este não é um entrave para que práticas/cuidados relacionados a esta nova terminologia sejam utilizadas durante a assistência à parturiente. De acordo com os objetivos propostos, conseguiu-se determinar os critérios e efeitos esperados pelas enfermeiras obstétricas ao utilizarem as tecnologias não-invasivas de cuidado estudadas, entretanto também se evidenciou brechas no conhecimento científico. Assim conclui-se que as enfermeiras obstétricas utilizam práticas/cuidados relacionadas as tecnologias não-invasivas de cuidado de enfermagem obstétrica no suporte físico à parturiente, pautadas em critérios e efeitos esperados que em sua maioria possuem bases científicas que os comprove. Considera-se que a utilização destas são uma ferramenta importante para a desmedicalização do processo de parto e consequentemente, para a diminuição dos índices de morbimortalidade materna e neonatall. Portanto, é necessário estimular à assistência ao parto por enfermeiras obstétricas, de modo a suplantar o modelo de assistência tecnocrático, ainda hegemônico no país.

I. Studies on bacteriophage DNA's and minicircular DNA's in M. lysodeikticus and E. coli 15. II. Flow dichroism of DNA solutions

Part I

Chapter 1.....A physicochemical study of the DNA molecules from the three bacteriophages, N1, N5, and N6, which infect the bacterium, M. lysodeikticus, has been made. The molecular weights, as measured by both electron microscopy and sedimentation velocity, are 23 x 10⁶ for N5 DNA and 31 x 10⁶ for N1 and N6 DNA's. All three DNA's are capable of thermally reversible cyclization. N1 and N6 DNA's have identical or very similar base sequences as judged by membrane filter hybridization and by electron microscope heteroduplex studies. They have identical or similar cohesive ends. These results are in accord with the close biological relation between N1 and N6 phages. N5 DNA is not closely related to N1 or N6 DNA. The denaturation T_m of all three DNA's is the same and corresponds to a (GC) content of 70%. However, the buoyant densities in CsCl of Nl and N6 DNA's are lower than expected, corresponding to predicted GC contents of 64 and 67%. The buoyant densities in Cs₂SO₄ are also somewhat anomalous. The buoyant density anomalies are probably due to the presence of odd bases. However, direct base composition analysis of N1 DNA by anion exchange chromatography confirms a GC content of 70%, and, in the elution system used, no peaks due to odd bases are present.

Chapter 2.....A covalently closed circular DNA form has been observed as an intracellular form during both productive and abortive infection processes in M. lysodeikticus. This species has been isolated by the method of CsC1-ethidium bromide centrifugation and examined with an electron microscope.

Chapter 3.....A minute circular DNA has been discovered as a homogeneous population in M. lysodeikticus. Its length and molecular weight as determined by electron microscopy are 0.445 μ and 0.88 x 10⁶ daltons respectively. There is about one minicircle per bacterium.

Chapter 4.....Several strains of E. coli 15 harbor a prophage. Viral growth can be induced by exposing the host to mitomycin C or to uv irradiation. The coliphage 15 particles from E. coli 15 and E, coli 15 T^- appear as normal phage with head and tail structure; the particles from E. coli 15 TAU are tailless. The complete particles exert a colicinogenic activity on E.coli 15 and 15 T^-, the tailless particles do not. No host for a productive viral infection has been found and the phage may be defective. The properties of the DNA of the virus have been studied, mainly by electron microscopy. After induction but before lysis, a closed circular DNA with a contour length of about 11.9 μ is found in the bacterium; the mature phage DNA is a linear duplex and 7.5% longer than the intracellular circular form. This suggests the hypothesis that the mature phage DNA is terminally repetitious and circularly permuted. The hypothesis was confirmed by observing that denaturation and renaturation of the mature phage DNA produce circular duplexes with two single-stranded branches corresponding to the terminal repetition. The contour length of the mature phage DNA was measured relative to φX RFII DNA and λ DNA; the calculated molecular weight is 27 x 10⁶. The length of the single-stranded terminal repetition was compared to the length of φX 174 DNA under conditions where single-stranded DNA is seen in an extended form in electron micrographs. The length of the terminal repetition is found to be 7.4% of the length of the nonrepetitious part of the coliphage 15 DNA. The number of base pairs in the terminal repetition is variable in different molecules, with a fractional standard deviation of 0.18 of the average number in the terminal repetition. A new phenomenon termed "branch migration" has been discovered in renatured circular molecules; it results in forked branches, with two emerging single strands, at the position of the terminal repetition. The distribution of branch separations between the two terminal repetitions in the population of renatured circular molecules was studied. The observed distribution suggests that there is an excluded volume effect in the renaturation of a population of circularly permuted molecules such that strands with close beginning points preferentially renature with each other. This selective renaturation and the phenomenon of branch migration both affect the distribution of branch separations; the observed distribution does not contradict the hypothesis of a random distribution of beginning points around the chromosome.

Chapter 5....Some physicochemical studies on the minicircular DNA species in E. coli 15 (0.670 μ, 1.47 x 10⁶ daltons) have been made. Electron microscopic observations showed multimeric forms of the minicircle which amount to 5% of total DNA species and also showed presumably replicating forms of the minicircle. A renaturation kinetic study showed that the minicircle is a unique DNA species in its size and base sequence. A study on the minicircle replication has been made under condition in which host DNA synthesis is synchronized. Despite experimental uncertainties involved, it seems that the minicircle replication is random and the number of the minicircles increases continuously throughout a generation of the host, regardless of host DNA synchronization.

Part II

The flow dichroism of dilute DNA solutions (A₂₆₀≈0.1) has been studied in a Couette-type apparatus with the outer cylinder rotating and with the light path parallel to the cylinder axis. Shear gradients in the range of 5-160 sec.^-1 were studied. The DNA samples were whole, "half," and "quarter" molecules of T4 bacteriophage DNA, and linear and circular λb₂b₅c DNA. For the linear molecules, the fractional flow dichroism is a linear function of molecular weight. The dichroism for linear A DNA is about 1.8 that of the circular molecule. For a given DNA, the dichroism is an approximately linear function of shear gradient, but with a slight upward curvature at low values of G, and some trend toward saturation at larger values of G. The fractional dichroism increases as the supporting electrolyte concentration decreases.

Bases fisiológicas del crecimiento diferencial en especies de almeja del género Ruditapes: estudio de alometrías

La coexistencia de las especies R. philippinarum y R. decussatus en la costa atlántica, empleando los mismos recursos y con tasas de crecimiento diferenciadas, parece indicio de estrategias fisiológicas diferentes. Dichas estrategias pueden identificarse mediante la cuantificación de parámetros fisiológicos que intervienen en el crecimiento, el cual es resultante del balance energético. En el presente estudio, se ha hecho una aproximación a ello, eligiendo como parámetros a analizar, la tasa de aclaramiento (componente de ganancia energética) y la tasa metabólica (componente de pérdida energética). El crecimiento diferencial viene determinado por diferentes combinaciones de comportamientos fisiológicos, recogidos en modelos fisiológicos de crecimiento (modelo de adquisición de energía, de limitación de costes basales y de eficiencia metabólica). Con el objetivo de realizar una comparación inter-específica entre ambas especies, se ha estudiado además la relación alométrica de estos parámetros con el tamaño, ya que la distribución de tamaños es una característica de la población y constituye la base sobre la que se construyen curvas de crecimiento. De acuerdo a informes emitidos por la FAO, R. philippinarum presenta mayores tasas de crecimiento por lo que cabría esperar mayor tasa de aclaramiento y menor tasa metabólica en esta especie. Sin embargo los resultados no se corresponden con ello, siendo la tasa de aclaramiento mayor para R. decussatus y no encontrándose diferencias en la tasa metabólica. Las relaciones alométricas tampoco difieren entre especies. No se puede asociar la mayor tasa de crecimiento de R. philippinarum a ninguno de los modelos de crecimiento, ya que las diferencias inter-específicas en los parámetros estudiados son nulas o contrarias a lo esperado. La falta de disponibilidad de semillas de la especie R. decussatus para este estudio limita en parte estas conclusiones, ya que los patrones de crecimiento diferencial pueden diferir profundamente entre estadíos juvenil y adulto. Alternativamente las anomalías comentadas podrían explicarse por la presencia de híbridos de ambas especies entre los ejemplares estudiados y donde la heterosis del crecimiento podría disipar las diferencias entre especies.

Part I. Coenzyme B12 as a hydroformylation-type catalyst. Part II. Mechanisms of hydrogen transfer in the methylmalonyl coenzyme a mutase reaction

Part I

The mechanism of the hydroformylation reaction was studied. Using cobalt deuterotetracarbonyl and 1-pentene as substrates, the first step in the reaction, addition of cobalt tetracarbonyl to an olefin, was shown to be reversible.

Part II

The role of coenzyme B₁₂ in the isomerization of methylmalonyl coenzyme A to succinyl coenzyme A by methylmalonyl coenzyme A mutase was studied. The reaction was allowed to proceed to partial completion using a mixture of methylmalonyl coenzyme A and 4, 4, 4-tri-²H-methylmalonyl coenzyme A as substrate. The deuterium distribution in the product, succinyl coenzyme A, was shown to best fit a model in which hydrogen is transferred from C-4 of methylmalonyl coenzyme A to C-5’ of the adenosyl moiety of coenzyme B₁₂ in the rate determining step. The three hydrogens at the 5’-adenosyl position of the coenzyme B₁₂ intermediate are then able to become enzymatically equivalent before hydrogen is transferred from the coenzyme B₁₂ intermediate to form succinyl coenzyme A.

Proton magnetic resonance studies of ribonucleic acid complexes. I. Complexes of biological bases and oligonucleotides with RNA. II. Template recognition and the degeneracy of the genetic code

Part I. Complexes of Biological Bases and Oligonucleotides with RNA

The physical nature of complexes of several biological bases and oligonucleotides with single-stranded ribonucleic acids have been studied by high resolution proton magnetic resonance spectroscopy. The importance of various forces in the stabilization of these complexes is also discussed.

Previous work has shown that purine forms an intercalated complex with single-stranded nucleic acids. This complex formation led to severe and stereospecific broadening of the purine resonances. From the field dependence of the linewidths, T₁ measurements of the purine protons and nuclear Overhauser enhancement experiments, the mechanism for the line broadening was ascertained to be dipole-dipole interactions between the purine protons and the ribose protons of the nucleic acid.

The interactions of ethidium bromide (EB) with several RNA residues have been studied. EB forms vertically stacked aggregates with itself as well as with uridine, 3'-uridine monophosphate and 5'-uridine monophosphate and forms an intercalated complex with uridylyl (3' → 5') uridine and polyuridylic acid (poly U). The geometry of EB in the intercalated complex has also been determined.

The effect of chain length of oligo-A-nucleotides on their mode of interaction with poly U in D₂0 at neutral pD have also been studied. Below room temperatures, ApA and ApApA form a rigid triple-stranded complex involving a stoichiometry of one adenine to two uracil bases, presumably via specific adenine-uracil base pairing and cooperative base stacking of the adenine bases. While no evidence was obtained for the interaction of ApA with poly U above room temperature, ApApA exhibited complex formation of a 1:1 nature with poly U by forming Watson-Crick base pairs. The thermodynamics of these systems are discussed.

Part II. Template Recognition and the Degeneracy of the Genetic Code

The interaction of ApApG and poly U was studied as a model system for the codon-anticodon interaction of tRNA and mRNA in vivo. ApApG was shown to interact with poly U below ~20°C. The interaction was of a 1:1 nature which exhibited the Hoogsteen bonding scheme. The three bases of ApApG are in an anti conformation and the guanosine base appears to be in the lactim tautomeric form in the complex.

Due to the inadequacies of previous models for the degeneracy of the genetic code in explaining the observed interactions of ApApG with poly U, the "tautomeric doublet" model is proposed as a possible explanation of the degenerate interactions of tRNA with mRNA during protein synthesis in vivo.

Theoretical research on noncollinear match conditions of the type I optical parametric process

Broad bandwidth group match conditions are reported for a noncollinear type I optical parametric process. The theoretical calculations corresponding to two special situations in practice were made, respectively, which are in accordance with the published experimental results. Furthermore, we provide a method to not only achieve maximal parametric bandwidth output but also match the group velocities between three waves. (c) 2006 Optical Society of America.

Tratamento da incontinência urinária de esforço por meio de eletroestimulação funcional dos músculos do assoalho pélvico: revisão sistemática de ensaios clínicos aleatorizados

Este estudo teve como objetivo verificar, através de uma revisão sistemática de ensaios clínicos aleatorizados, os benefícios da estimulação elétrica funcional endovaginal ou dos tratamentos conservadores às pacientes com incontinência urinária de esforço, e demonstrar qual modalidade de tratamento conservador apresenta melhores resultados na terapêutica dessas mulheres: a estimulação elétrica funcional endovaginal, em comparação com os cones vaginais ou a realização de exercícios perineais. Para tanto, foram realizadas buscas nas principais bases de dados científicos, por estudos que atendessem a pergunta da pesquisa, tipo de intervenção e tipo de participantes selecionados. Destes, foram selecionados 7 estudos que foram submetidos à análise dos revisores, que avaliaram os seguintes desfechos: episódios de perda urinária, quantificação das perdas urinárias através do pad-test, força da musculatura perineal, qualidade de vida, volume residual, capacidade cistométrica máxima, melhora dos sintomas, satisfação e cura. Todas as terapias pesquisadas apresentaram melhora dos sintomas da incontinência urinária de esforço; no entanto, segundo os desfechos avaliados, apresentaram diferença no resultado comparativo. Quanto às perdas urinárias, ao pad-test e à força da musculatura perineal, a realização dos exercícios pélvicos obteve os melhores resultados. Já a terapia por estimulação elétrica endovaginal e a terapia com os cones apresentaram resultados semelhantes, não sendo encontrada diferença significativa em nenhum dos desfechos analisados. De acordo com os achados obtidos nesta revisão sistemática, entendemos que o tratamento pela estimulação elétrica traz benefícios às pacientes com incontinência urinária de esforço. Os exercícios pélvicos demonstraram ser a terapia que reduz mais significativamente os sintomas ocasionados por esta condição

Optical setup and analysis of disk-type photopolymer high-density holographic storage

A relatively simple scheme for disk-type photopolymer high-density holographic storage based on angular and spatial multiplexing is described. The effects of the optical setup on the recording capacity and density are studied. Calculations and analysis show that this scheme is more effective than a scheme based on the spatioangular multiplexing for disk-type photopolymer high-density holographic storage, which has a limited medium thickness. Also an optimal beam recording angle exists to achieve maximum recording capacity and density. (C) 2002 Society of Photo-Optical Instrumentation Engineers.

Analysis on a new type magneto-optical disk for blue lasers

The conventional TbFeCo magneto-optical (MO) medium has a relatively smaller Kerr rotation angle in the blue region than in the red. With the recording wavelength gradually moving to the short wavelength, if TbFeCo is still used as recording medium, the conventional MO disk structure must be optimized to get a larger carrier to noise ratio (CNR). Sabi et al. have found that adding a metal layer attached to the TbFeCo film as thermal control layer is a useful way to get a high CNR. In this paper, we proved this through calculation, and carried out optimization of the new type of disk. Calculation results showed that the new structure is useful in preventing an excessive temperature increase, and has a better thermal response. (c) 2005 Elsevier B.V. All rights reserved.

Intervenções psicossociais para Transtornos Mentais Comuns na Atenção Primária à Saúde.

Os Transtornos Mentais Comuns (TMC), especialmente ansiedade e depressão, são altamente prevalentes, independentemente das dificuldades com a sua classificação. Na Atenção Primária à Saúde (APS), representada no Brasil pela Estratégia de Saúde da Família (ESF), essas condições são pouco detectadas e tratadas, o que resulta em sobrecarga para pacientes, famílias e comunidades, além de prejuízos sociais e econômicos. Tal situação pode ser atenuada integrando-se cuidados de saúde mental à atenção básica. Nesse sentido, objetivou-se identificar e analisar as intervenções psicossociais voltadas para o cuidado do sofrimento psíquico e dos TMC atualmente empregadas na APS, no Brasil e no exterior. Para isso, foi realizada uma revisão bibliográfica da literatura nacional e internacional através das bases de dados MEDLINE/PubMed e LILACS/BVS. As intervenções identificadas foram reunidas em dois grupos. No primeiro, incluíram-se as intervenções com formato tradicional em que um profissional graduado conduz o tratamento presencialmente, enquanto no segundo grupo foram alocados os arranjos diferenciados em que o contato com o profissional é reduzido e seu papel é mais o de facilitar do que de liderar o tratamento, tal como nas intervenções com o uso de mão de obra leiga em saúde, suporte ao autocuidado, e sistemas automatizados ou pacotes de tratamento informatizados oferecidos via internet. Teoricamente, Terapia Interpessoal, Terapia Cognitivo-comportamental e Terapia de Solução de Problemas embasaram as intervenções. Na APS internacional, as intervenções são breves e bastante estruturadas, incluindo um plano de ação definido em um manual de aplicação. Na Estratégia de Saúde da Família destacaram-se as intervenções grupais e a Terapia Comunitária, modelo especialmente desenvolvido por brasileiros. Verificou-se que os TMC ganham visibilidade à medida que a APS se consolida no território e próxima à população. No exterior, a integração da saúde mental na APS é um processo em consolidação e há crescente tecnificação das intervenções psicossociais, enquanto, no Brasil, a aproximação da APS com a ESF é recente e não há um padrão de atuação consolidado. Resultados insuficientes ou controversos impediram concluir quais são as melhores intervenções, porém a alta prevalência dos TMC e o potencial de cuidado do setor da APS exigem que as pesquisas continuem.