Persistência de plasmídeos que codificam carbapenemases do tipo New-Delhi-Metalo-β-Lactamase

As metalo-β-lactamases (MBL) são capazes de hidrolisar os carbapenêmicos, a classe de antimicrobianos com maior potência para o tratamento de infecções graves e de maior uso clinico. Dentre as MBL, o grupo mais recentemente descrito e que apresentou rápida disseminação em todo o mundo é o da New-Delhi-Metalo- β-lactamases (NDM). Nas enterobactérias, os genes que codificam essas enzimas estão mais frequentemente localizados em plasmídeos. O estudo da estabilidade de plasmídeos que albergam o gene blaNDM-1 é importante para entender a predominância de espécies que carregam esses plasmídeos, desvendar mecanismos moleculares envolvidos na sua persistência e para desenvolver novas drogas que possam diminuir a sua persistência. Estudos recentes sobre estabilidade plasmidial evidenciaram que a maprotilina é capaz de induzir perda plasmidial de até 90% em E. coli K12. Neste trabalho, foi estudado o efeito da maprotilina na indução de cura de plasmídeos, que albergam o gene blaNDM-1, em diferentes espécies da família Enterobacteriaceae. Nove isolados pertencentes a diferentes espécies foram incluídas no estudo. Os plasmídeos foram caracterizados quanto ao seu tamanho por eletroforese e por sequenciamento de DNA no sistema Illumina. A persistência plasmidial foi determinada pelo método de contagem em placa em LB ágar com e sem tratamento com maprotilina em concentrações sub-inibitórias (50mg/L). O experimento foi conduzido por 10 dias, representando aproximadamente 100 gerações. Neste estudo evidenciou-se que o grupo das enterobactérias estão envolvidas na disseminação de plasmídeos com blaNDM-1, sendo que plasmídeos do grupo IncF estão mais relacionados a essa dispersão. A maprotilina teve efeito de cura plasmidial em todos os isolados exceto em E. hormaechei \"subsp. oharae\" e C. freundii. O isolado P. rettgeri apresentou maior taxa de perda plasmidial e a análise comparativa da sequência nucleotídica do plasmídeo indicou que a presença da IS5 pode estar relacionada com a diminuição da persistência plasmidial. Diferenças na persistência plasmidial, quando tratados com maprotilina, entre E. hormaechei \"subsp. steigerwaltii\" e E. hormaechei \"subsp. oharae\" sugerem que E. hormaechei \"subsp. oharae\" pode ser um possível disseminador de plasmídeos albergando blaNDM-1, devido a processos de adaptação co-evolutivos.

Écoulements granulaires par avalanches : indices de fluidité, fractales et multifractales

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Écoulements granulaires par avalanches : indices de fluidité, fractales et multifractales

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Lithic grain and mineral composition of sand and sandstones from the North Pacific Ocean and the Bering Sea (Table 3)

Sand and sandstone compositions from different types of basins reflect provenance terranes governed by plate tectonics. One hundred and one thin sections of Upper Miocene to Holocene sand-sized material were examined from DSDP/IPOD Sites in the North Pacific Ocean and the Bering Sea. The Gazzi-Dickinson point-counting method was used to establish compositional characteristics of sands from different tectonic settings. Continental margin forearc sands from the western North America continental margin arc system are clearly different from backarc/marginal-sea sands from the Aleutian intraoceanic arc system. The forearc sands have average QFL percentages of 29-42-29, LmLvLst percentages of 32-34-34, 3 Fmwk%M and 0.82 P/F. Aleutian backarc sands have average QFL percentages of 8-22-69. LmLvLst percentages of 9-85-6, 0.5 Fmwk%M and 0.96 P/F. A trend of increasing QFL%Q and decreasing LmLvLst%Lv westward in the backarc region of the Aleutian Ridge reflects the influence of the Asiatic continental margin. Aleutian backarc sands without continental influence have average QFL percentages of 1-20-79, LmLvLst percentages of 1-98-1, 0 Fmwk%M and 0.99 P/F. Of the continental margin forearc samples, sands on the Astoria Fan (west of the Oregon-Washington trench) contain the highest LmLvLst%Lv and lowest P/F; sands from mixed transform-fault and trench settings (Delgada Fan and Gulf of Alaska samples) have slightly higher Qp/Q (0.03); and sands from the Pacific-Juan de Fuca-North America triple junction have the highest Fmwk%M. Delgada Fan and Gulf of Alaska sands have average QFL percentages of 27-38-35, LmLvLst percentages of 37-26-37, 2 Fmwk%M and 0.86 P/F. Astoria Fan sands have average QFL percentages of 35-41-24, LmLvLst percentages of 30-47-23, 3 Fmwk%M and 0.74 P/F. The triple-junction sands have average QFL percentages of 28-59-13, LmLvLst percentages of 25-26-49, 9 Fmwk%M and 0.87 P/F. The petrologic data from the modern ocean basins examined in this study can provide useful analogs for interpretation of ancient oceanic sequences. Our data suggest some refinements of, but generally substantiate, existing petrologic models relating sandstone composition to tectonic setting.

Avaliação da atividade antimicrobiana de filmes protéicos a base de anchoita (engraulis anchoita) incorporados com ácidos orgânicos.

Os filmes são produzidos a partir de macromoléculas, que podem ser utilizados como embalagem, como os polissacarídeos, lipídeos e proteínas. As proteínas se destacam dos demais, pois possuem uma estrutura com 20 monômeros diferentes, que confere um amplo potencial de ligações intermoleculares. A incorporação de agentes ativos em filmes é uma alternativa como embalagem, para inibir ou retardar a multiplicação de microrganismos patógenos e deteriorantes em alimentos. O objetivo deste trabalho foi avaliar a atividade antimicrobiana de filmes à base de isolado protéico de anchoita (Engraulis anchoita) – IPA adicionados de ácidos orgânicos. Para tanto, foi elaborado o IPA, pela solubilização alcalina da proteína e precipitação no ponto isoelétrico a partir de carne mecanicamente separada. O IPA foi avaliado quanto a sua composição proximal, aminoacídica e por DSC. A solução formadora dos filmes foi elaborada a partir de IPA, água, glicerol e hidróxido de sódio. As formulações dos filmes foram elaboradas segundo um planejamento fatorial 23 . Foram avaliadas as propriedades físico-químicas de resistência a tração (RT) e elongação (E); espessura, solubilidade e permeabilidade ao vapor de água (PVA); a diferença de cor (∆E*) e opacidade (Y) e microscopia eletrônica de varredura (MEV) de filmes à base de IPA. Os filmes com diferentes concentrações de ácido sórbico (AS) ou ácido benzóico (AB) foram desenvolvidos a partir da condição cujo as propriedades físico-químicas foram as melhores, sendo comparados aos filmes controles. Estes, foram avaliados quanto a sua atividade antimicrobiana frente aos microrganismos Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus e Salmonella Enteritidis pelo método de difusão em disco, além das propriedades físico-químicas, MEV e FT-IV. Os filmes com maior atividade antimicrobiana e os filmes controle foram aplicados sobre carne bovina, inoculados com os microrganismos inibidos no método de difusão em disco e armazenados a 5°C. Estes, foram avaliados a cada 2 dias durante 12 dias de armazenamento, pela método de contagem em gotas. O IPA apresentou 88,8% de proteína e 53,3% de aminoácidos polares e temperatura de desnaturação de 62,2°C. A espessura, PVA, ∆E* e Y dos filmes não foram afetados pelas variáveis estudadas no experimento. A menor solubilidade e maior RT dos filmes ocorreram em baixa concentração de IPA, glicerol e tratamento térmico, mas a E aumentou com o acréscimo dessas variáveis. As MEV das superfícies dos filmes foram homogêneas, para aqueles com leve tratamento térmico. O aumento da concentração de AS e AB na faixa de 0,50 a 1,50% resultou na diminuição da RT e aumento da E, solubilidade, ∆E* e Y. Houve mudança da organização molecular e interações intermoleculares entre as moléculas de IPA e AB testados pela avaliação do FT-IV. As MEV revelaram microporos em filmes com 1,50% de AS, o que resultou em filmes com menor homogeneidade. A maior atividade antimicrobiana foi verificada nos filmes com 1,50% de AS e AB frente a E. coli O157:H7, L. monocytogenes e S. Enteritidis. Estes filmes foram aplicados sobre carne bovina inoculada com E. coli O157:H7 e L. monocytogenes. Os filmes de AS frente a E. coli O157:H7 e L. monocytogenes apresentaram uma redução de 5 e 4 log UFC.g-1, respectivamente, em relação ao filme controle. O efeito do AB frente a estas bactérias, apresentou uma redução de 6 e 5 log UFC.g-1, ao final do 12° dia de armazenamento, respectivamente. Os filmes elaborados à base de IPA, adicionados de AS ou AB podem ser eficazes contra os patógenos alimentares testados.

Bayesian Linear Regression for estimation of the Fractal Dimension of the Cerebral Cortex

The cerebral cortex presents self-similarity in a proper interval of spatial scales, a property typical of natural objects exhibiting fractal geometry. Its complexity therefore can be characterized by the value of its fractal dimension (FD). In the computation of this metric, it has usually been employed a frequentist approach to probability, with point estimator methods yielding only the optimal values of the FD. In our study, we aimed at retrieving a more complete evaluation of the FD by utilizing a Bayesian model for the linear regression analysis of the box-counting algorithm. We used T1-weighted MRI data of 86 healthy subjects (age 44.2 ± 17.1 years, mean ± standard deviation, 48% males) in order to gain insights into the confidence of our measure and investigate the relationship between mean Bayesian FD and age. Our approach yielded a stronger and significant (P < .001) correlation between mean Bayesian FD and age as compared to the previous implementation. Thus, our results make us suppose that the Bayesian FD is a more truthful estimation for the fractal dimension of the cerebral cortex compared to the frequentist FD.

Frattali autosimili e misura di Hausdorff

In questa tesi si vuole trattare il concetto di dimensione a partire dalla teoria della misura e lo si vuole applicare per definire e studiare gli insiemi frattali, in particolare, autosimili. Nel primo capitolo si tratta la dimensione di Hausdorff a partire dalla teoria della misura di Hausdorff, di cui si osservano alcune delle proprietà grazie a cui la prima si può definire. Nel secondo capitolo si danno altre definizioni di dimensione, come ad esempio quella di auto-similarità e la box-counting, per mostrare che tale concetto non è univoco. Si analizzano quindi le principali differenze tra le diverse dimensioni citate e si forniscono esempi di insiemi per cui esse coincidono e altri, invece, per cui esse differiscono. Nel terzo capitolo si introduce poi il vero e proprio concetto di insieme frattale. In particolare, definendo i sistemi di funzioni iterate e studiandone le principali proprietà, si definisce una particolare classe di insiemi frattali detti autosimili. In questo capitolo sono enunciati e dimostrati teoremi fondamentali che legano gli attrattori di sistemi di funzioni iterate a insiemi frattali autosimili e forniscono, per alcuni specifici casi, una formula per calcolarne la dimensione di Hausdorff. Si danno, inoltre, esempi di calcolo di tale dimensione per alcuni insiemi frattali autosimili molto noti. Nel quarto capitolo si dà infine un esempio di funzione che abbia grafico frattale, la Funzione di Weierstrass, per mostrare un caso pratico in cui è utilizzata la teoria studiata nei capitoli precedenti.

Box-Behnken design for the optimization of an enantioselective method for the simultaneous analysis of propranolol and 4-hydroxypropranolol by CE

An experimental design optimization (Box-Behnken design, BBD) was used to develop a CE method for the simultaneous resolution of propranolol (Prop) and 4-hydroxypropranolol enantiomers and acetaminophen (internal standard). The method was optimized using an uncoated fused silica capillary, carboxymethyl-beta-cyclodextrin (CM-beta-CD) as chiral selector and triethylamine/phosphoric acid buffer in alkaline conditions. A BBD for four factors was selected to observe the effects of buffer electrolyte concentration, pH, CM-beta-CD concentration and voltage on separation responses. Each factor was studied at three levels: high, central and low, and three center points were added. The buffer electrolyte concentration ranged from 25 to 75 mM, the pH ranged from 8 to 9, the CM-beta-CD concentration ranged from 3.5 to 4.5%w/v, and the applied run voltage ranged from 14 to 20 W. The responses evaluated were resolution and migration time for the last peak. The obtained responses were processed by Minitab (R) to evaluate the significance of the effects and to find the optimum analysis conditions. The best results were obtained using 4%w/v CM-beta-CD in 25 mM triethylamine/H(3)PO(4) buffer at pH 9 as running electrolyte and 17 kV of voltage. Resolution values of 1.98 and 1.95 were obtained for Prop and 4-hydroxypropranolol enantiomers, respectively. The total analysis time was around of 15 min. The BBD showed to be an adequate design for the development of a CE method, resulting in a rapid and efficient optimization of the pH and concentration of the buffer, cyclodextrin concentration and applied voltage.

Evaluation of the PetrifilmTM and TEMPO® systems and the conventional method for counting microorganisms in pasteurized milk

New microbiological methods have been developed and commercialized, but their performance must be guaranteed. The aim of the present study was to evaluate the PetrifilmTM and TEMPO® systems compared to the conventional method for counting microorganisms in pasteurized milk. A total of 141 samples of pasteurized milk were analyzed by counting mesophilic aerobic, Coliforms at 35 ºC, Coliforms at 45 ºC, and Escherichia coli microorganisms. High correlation was found between the methods for counting Coliforms at 35 ºC, but low correlation was found for counting mesophilic aerobic, Coliforms at 45 ºC, and Escherichia coli. No significant statistical difference was found among the three methods for counting Coliforms at 35 ºC; however, the mean counts of mesophilic aerobic, Coliforms at 45 ºC, and Escherichia coli showed significant statistical difference. PetrifilmTM and TEMPO® systems had satisfactory results for Coliforms at 35 ºC in pasteurized milk but low performance for mesophilic aerobic, Coliforms at 45 ºC and Escherichia coli.

Use of an Inverse Method for Time Series to Estimate the Dynamics of and Management Strategies for the Box Jellyfish Carybdea marsupialis

Frequently, population ecology of marine organisms uses a descriptive approach in which their sizes and densities are plotted over time. This approach has limited usefulness for design strategies in management or modelling different scenarios. Population projection matrix models are among the most widely used tools in ecology. Unfortunately, for the majority of pelagic marine organisms, it is difficult to mark individuals and follow them over time to determine their vital rates and built a population projection matrix model. Nevertheless, it is possible to get time-series data to calculate size structure and densities of each size, in order to determine the matrix parameters. This approach is known as a “demographic inverse problem” and it is based on quadratic programming methods, but it has rarely been used on aquatic organisms. We used unpublished field data of a population of cubomedusae Carybdea marsupialis to construct a population projection matrix model and compare two different management strategies to lower population to values before year 2008 when there was no significant interaction with bathers. Those strategies were by direct removal of medusae and by reducing prey. Our results showed that removal of jellyfish from all size classes was more effective than removing only juveniles or adults. When reducing prey, the highest efficiency to lower the C. marsupialis population occurred when prey depletion affected prey of all medusae sizes. Our model fit well with the field data and may serve to design an efficient management strategy or build hypothetical scenarios such as removal of individuals or reducing prey. TThis This sdfsdshis method is applicable to other marine or terrestrial species, for which density and population structure over time are available.

Improved method of detecting and counting bacteria. /

Address inquiries and all applications for license for this invention to NASA Patent Counsel, Goddard Space Flight Center, Mail Code 204, Greenbelt, Maryland, 20771.

Cu2ZnSnS4 absorber layers obtained through sulphurization of metallic precursors: Graphite box versus sulphur flux

In this work we employed a hybrid method, combining RF-magnetron sputtering with evaporation, for the deposition of tailor made metallic precursors, with varying number of Zn/Sn/Cu (ZTC) periods and compared two approaches to sulphurization. Two series of samples with 1×, 2× and 4× ZTC periods have been prepared. One series of precursors was sulphurized in a tubular furnace directly exposed to a sulphur vapour and N2+5% H2 flux at a pressure of 5.0×10+4 Pa. A second series of identical precursors was sulphurized in the same furnace but inside a graphite box where sulphur pellets have been evaporated again in the presence of N2+5% H2 and at the same pressure as for the sulphur flux experiments. The morphological and chemical analyses revealed a small grain structure but good average composition for all three films sulphurized in the graphite box. As for the three films sulphurized in sulphur flux grain growth was seen with the increase of the number of ZTC periods whilst, in terms of composition, they were slightly Zn poor. The films' crystal structure showed that Cu2ZnSnS4 is the dominant phase. However, in the case of the sulphur flux films SnS2 was also detected. Photoluminescence spectroscopy studies showed an asymmetric broad band emission whichoccurs in the range of 1–1.5 eV. Clearly the radiative recombination efficiency is higher in the series of samples sulphurized in sulphur flux. We have found that sulphurization in sulphur flux leads to better film morphology than when the process is carried out in a graphite box in similar thermodynamic conditions. Solar cells have been prepared and characterized showing a correlation between improved film morphology and cell performance. The best cells achieved an efficiency of 2.4%.

Optimisation and design of post-tensioning anchorage corner blisters in concrete box girder bidges

The design of anchorage blisters of internal continuity post-tensioning tendons of bridges built by the cantilever method, presents some peculiarities, not only because they are intermediate anchorages but also because these anchorages are located in blisters, so the prestressing force has to be transferred from the blister the bottom slab and web of the girder. The high density of steel reinforcement in anchorage blisters is the most common reason for problems with concrete cast in situ, resulting in zones with low concrete compacity, leading to concrete crushing failures under the anchor plates. A solution may involve improving the concrete compression and tensile strength. To meet these requirements a high-performance fibre reinforced self-compacting mix- ture (HPFRC) was used in anchorage corner blisters of post-tensioning tendons, reducing the concrete cross-section and decreasing the reinforcement needed. To assess the ultimate capacity and the adequate serviceability of the local anchorage zone after reducing the minimum concrete cross-section and the confining reinforcement, specified by the anchorage device supplier for the particular tendon, load transfer tests were performed. To investigate the behaviour of anchorage blisters regarding the transmission of stresses to the web and the bottom slab of the girder, and the feasibility of using high performance concrete only in the blister, two half scale models of the inferior corner of a box girder existing bridge were studied: a reference specimen of ordinary reinforced concrete and a HPFRC blister specimen. The design of the reinforcement was based in the tensile forces obtained on strut-and-tie models. An experimental program was carried out to assess the models used in design and to study the feasibility of using high performance concrete only in the blister, either with casting in situ, or with precast solutions. A non-linear finite element analysis of the tested specimens was also performed and the results compared.