Optimization of peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) for the detection of bacteria: the effect of pH, dextran sulfate and probe concentration

Fluorescence in situ hybridization (FISH) is a molecular technique widely used for the detection and characterization of microbial populations. FISH is affected by a wide variety of abiotic and biotic variables and the way they interact with each other. This is translated into a wide variability of FISH procedures found in the literature. The aim of this work is to systematically study the effects of pH, dextran sulfate and probe concentration in the FISH protocol, using a general peptide nucleic acid (PNA) probe for the Eubacteria domain. For this, response surface methodology was used to optimize these 3 PNA-FISH parameters for Gram-negative (Escherichia coli and Pseudomonas fluorescens) and Gram-positive species (Listeria innocua, Staphylococcus epidermidis and Bacillus cereus). The obtained results show that a probe concentration higher than 300 nM is favorable for both groups. Interestingly, a clear distinction between the two groups regarding the optimal pH and dextran sulfate concentration was found: a high pH (approx. 10), combined with lower dextran sulfate concentration (approx. 2% [w/v]) for Gram-negative species and near-neutral pH (approx. 8), together with higher dextran sulfate concentrations (approx. 10% [w/v]) for Gram-positive species. This behavior seems to result from an interplay between pH and dextran sulfate and their ability to influence probe concentration and diffusion towards the rRNA target. This study shows that, for an optimum hybridization protocol, dextran sulfate and pH should be adjusted according to the target bacteria.

Understanding adaptation to pH in an industrially relevant glycoside hydrolase family 8 xylanase

Dissertação de mestrado em Genética Molecular

Engineering a commercial cold-adapted xylanase for improved low pH stability

Dissertação de mestrado em Genética Molecular

Effect of pH on Candida tropicalis virulence factors

Dissertação de mestrado em Bioengenharia

Evaluation of the determinism of the actuator sensor interface (ASI) on programmable logic controllers (PLC), personal computers (PC) and real-time operating systems (RTOS)

The purpose of this study was to evaluate the determinism of the AS-lnterface network and the 3 main families of control systems, which may use it, namely PLC, PC and RTOS. During the course of this study the PROFIBUS and Ethernet field level networks were also considered in order to ensure that they would not introduce unacceptable latencies into the overall control system. This research demonstrated that an incorrectly configured Ethernet network introduces unacceptable variable duration latencies into the control system, thus care must be exercised if the determinism of a control system is not to be compromised. This study introduces a new concept of using statistics and process capability metrics in the form of CPk values, to specify how suitable a control system is for a given control task. The PLC systems, which were tested, demonstrated extremely deterministic responses, but when a large number of iterations were introduced in the user program, the mean control system latency was much too great for an AS-I network. Thus the PLC was found to be unsuitable for an AS-I network if a large, complex user program Is required. The PC systems, which were tested were non-deterministic and had latencies of variable duration. These latencies became extremely exaggerated when a graphing ActiveX was included in the control application. These PC systems also exhibited a non-normal frequency distribution of control system latencies, and as such are unsuitable for implementation with an AS-I network. The RTOS system, which was tested, overcame the problems identified with the PLC systems and produced an extremely deterministic response, even when a large number of iterations were introduced in the user program. The RTOS system, which was tested, is capable of providing a suitable deterministic control system response, even when an extremely large, complex user program is required.

Architektur für verteilte, fehlertolerante Sensor-Aktor-Systeme

Magdeburg, Univ., Fak. für Informatik, Diss., 2011

Quasi-integrals and the origin of bursting pH oscillations in an enzyme model system

Bursting Oscillation, Mixed-Mode oscillation, Slow Manifold, Quasi-Integral, slow-fast analysis, QSSA

Wideband impedance spectrum analyzer with arbitry fine frequency resolution for in situ sensor applications

Magdeburg, Univ., Fak. für Elektrotechnik und Informationstechnik, Diss., 2009

Steuerung eines zweibeinigen Schreitroboters mit einem elastischen Sensor-Aktor-System

Magdeburg, Univ., Fak. für Elektrotechnik und Informationstechnik, Diss., 2012

"O pH de soluções de ácido acético, cítrico, oxálico e tartárico"

O presente trabalho descreve os dados obtidos sôbre a determinação do pH em soluções, desde 0,005 até 0,50 molar de ácido acético, ácido cítrico, ácido oxálico e ácido tartárico. Os dados obtidos experimentalmente, quando expressos em função de pC, isto é, em função de log log 1/C apresentaram uma relação linear. Por outro lado, calculando-se o pH das diversas soluções dos ácidos estudados, através de duas equações, uma do primeiro grau e outra do segundo grau, observou-se que os resultados calculados pela segunda equação apresentaram valores muito próximos aos determinados experimentalmente, conquanto no cálculo tenha sido usada apenas, a primeira constante termodinâmica de ionização, para os ácido cítrico, oxálico e tartárico. Uma vez que o valor do pH determinado e o do pH calculado constituem uma função linear do pC, foram estabelecidas duas equações de regressão para cada ácido estudado. Na primeira equação de regressão o pH determinado figura como variável dependente e na segunda, o pH calculado é a variável dependente. Nas duas equações o pC é a variável independente.

Estudos sôbre a alimentação mineral do cafeeiro: XX. efeito da variação de pH no desenvolvimento e composição química do cafeeiro (Coffea arabica L., var. Mundo Nôvo) cultivado em solução nutritiva

Plantas jovens de Coffea arabica L., var. Mundo Nôvo foram cultivadas em solução nutritiva, sob o efeito de diferentes variações de pH (4,0 a 7,5) do substrato, a fim de se constatar o desenvolvimento e composição mineral. A melhor faixa de pH para o crescimento em altura, número de fôlhas, pêso da matéria fresca e sêca é de 4,0 a 6,0. A quantidade total de todos os macronutrientes absorvidos pelo cafeeiro diminui à medida que o pH se eleva.