LacO-LacI interaction in affinity adsorption of plasmid DNA

Current approaches for purifying plasmids from bacterial production systems exploit the physiochemical properties of nucleic acids in non-specific capture systems. In this study, an affinity system for plasmid DNA (pDNA) purification has been developed utilizing the interaction between the lac operon (lacO) sequence contained in the pDNA and a 64mer synthetic peptide representing the DNA-binding domain of the lac repressor protein, LacI. Two plasmids were evaluated, the native pUC19 and pUC19 with dual lacO3/lacOs operators (pUC19lacO3/lacOs), where the lacOs operator is perfectly symmetrical. The DNA-protein affinity interaction was evaluated by surface plasmon resonance using a Biacore system. The affinity capture of DNA in a chromatography system was evaluated using LacI peptide that had been immobilized to Streamline™ adsorbent. The KD-values for double stranded DNA (dsDNA) fragments containing lacO1 and lacO3 and lacOs and lacO3 were 5.7 ± 0.3 × 10 -11 M and 4.1 ± 0.2 × 10-11 M respectively, which compare favorably with literature reports of 5 × 10-10 - 1 × 10-9 M for native laCO1 and 1-1.2 × 10-10 M for lacO1 in a saline buffer. Densitometric analysis of the gel bands from the affinity chromatography run clearly showed a significant preference for capture of the supercoiled fraction from the feed pDNA sample. The results indicate the feasibility of the affinity approach for pDNA capture and purification using native protein-DNA interaction. © 2006 Wiley Periodicals, Inc.

Posterior retinal contour in adult human anisomyopia

PURPOSE. It is well documented that myopia is associated with an increase in axial length or, more specifically, in vitreous chamber depth. Whether the transverse dimensions of the eye also increase in myopia is relevant to further understanding of its development. METHODS. The posterior retinal surface was localized in two-dimensional space in both eyes of young adult white and Taiwanese-Chinese iso- and anisomyopes (N = 56), from measured keratometry, A-scan ultrasonography, and central and peripheral refraction (±35°) data, with the aid of a computer modeling program designed for this purpose. Anisomyopes had 2 D or more interocular difference in their refractive errors, with mean values in their more myopic eyes of -5.57 D and in their less myopic eyes of -3.25 D, similar to the means of the two isomyopic groups. The derived retinal contours for the more and less myopic eyes were compared by way of investigating ocular shape changes that accompany myopia, in the posterior region of the vitreous chamber. The presence and size of optic disc crescents were also investigated as an index of retinal stretching in myopia. RESULTS. Relative to the less myopic eyes of anisometropic subjects, the more myopic eyes were more elongated and also distorted into a more prolate shape in both the white and Chinese groups. However, the Chinese eyes showed a greater and more uniform relative expansion of the posterior retinal surface in their more myopic eyes, and this was associated with larger optic disc crescents. The changes in the eyes of whites displayed a nasal-temporal axial asymmetry, reflecting greater enlargement of the nasal retinal sector. CONCLUSIONS. Myopia is associated with increased axial length and a prolate shape. This prolate shape is consistent with the proposed idea that axial and transverse dimensions of the eye are regulated differently. The observations that ocular shape changes are larger but more symmetrical in Chinese eyes than in eyes of whites warrant further investigation.

A simple, sensitive and selective quantum-dot-based western blot method for the simultaneous detection of multiple targets from cell lysates

Quantum dots (Qdots) are fluorescent nanoparticles that have great potential as detection agents in biological applications. Their optical properties, including photostability and narrow, symmetrical emission bands with large Stokes shifts, and the potential for multiplexing of many different colours, give them significant advantages over traditionally used fluorescent dyes. Here, we report the straightforward generation of stable, covalent quantum dot-protein A/G bioconjugates that will be able to bind to almost any IgG antibody, and therefore can be used in many applications. An additional advantage is that the requirement for a secondary antibody is removed, simplifying experimental design. To demonstrate their use, we show their application in multiplexed western blotting. The sensitivity of Qdot conjugates is found to be superior to fluorescent dyes, and comparable to, or potentially better than, enhanced chemiluminescence. We show a true biological validation using a four-colour multiplexed western blot against a complex cell lysate background, and have significantly improved previously reported non-specific binding of the Qdots to cellular proteins.

Automated assessment of visual fields and their inter-relation to evoked potentials in visual disorders

The Octopus Automated Perimeter was validated in a comparative study and found to offer many advantages in the assessment of the visual field. The visual evoked potential was investigated in an extensive study using a variety of stimulus parameters to simulate hemianopia and central visual field defects. The scalp topography was recorded topographically and a technique to compute the source derivation of the scalp potential was developed. This enabled clarification of the expected scalp distribution to half field stimulation using different electrode montages. The visual evoked potential following full field stimulation was found to be asymmetrical around the midline with a bias over the left occiput particularly when the foveal polar projections of the occipital cortex were preferentially stimulated. The half field response reflected the distribution asymmetry. Masking of the central 3° resulted in a response which was approximately symmetrical around the midline but there was no evidence of the PNP-complex. A method for visual field quantification was developed based on the neural representation of visual space (Drasdo and Peaston 1982) in an attempt to relate visual field depravation with the resultant visual evoked potentials. There was no form of simple, diffuse summation between the scalp potential and the cortical generators. It was, however, possible to quantify the degree of scalp potential attenuation for M-scaled full field stimuli. The results obtained from patients exhibiting pre-chiasmal lesions suggested that the PNP-complex is not scotomatous in nature but confirmed that it is most likely to be related to specific diseases (Harding and Crews 1982). There was a strong correlation between the percentage information loss of the visual field and the diagnostic value of the visual evoked potential in patients exhibiting chiasmal lesions.

Engineering liposomal systems to develop solubility enhancing technology

This research primarily focused on identifying the formulation parameters which control the efficacy of liposomes as delivery systems to enhance the delivery of poorly soluble drugs. Preliminary studies focused on the drug loading of ibuprofen within vesicle systems. Initially both liposomal and niosomal formulations were screened for their drug-loading capacity: liposomal systems were shown to offer significantly higher ibuprofen loading and thereafter lipid based systems were further investigated. Given the key role cholesterol is known to play within the stability of bilayer vesicles. the optimum cholesterol content in terms of drug loading and release of poorly soluble drugs was then investigated. From these studies a concentration of 11 total molar % of cholesterol was used as a benchmark for all further formulations. Investigating the effect of liposomc composition on several low solubility drugs, drug loading was shown to be enhanced by adopting longer chain length lipids. cationic lipids and. decreasing drug molecular weight. Drug release was increased by using cationic lipids and lower molecular weight of drug; conversely, a reduction was noted when employing longer chain lipids thus supporting the rational of longer chain lipids producing more stable liposomes, a theory also supported by results obtained via Langmuir studies· although it was revealed that stability is also dependent on geometric features associated with the lipid chain moiety. Interestingly, reduction in drug loading appeared to be induced when symmetrical phospholipids were substituted for lipids constituting asymmetrical alkyl chain groups thus further highlighting the importance of lipid geometry. Combining a symmetrical lipid with an asymmetrical derivative enhanced encapsulation of a hydrophobic drug while reducing that of another suggesting the importance of drug characteristics. Phosphatidylcholine liposornes could successfully be prepared (and visualised using transmission electron microscopy) from fatty alcohols therefore offering an alternative liposomal stabiliser to cholesterol. Results obtained revealed that liposomes containing tetradecanol within their formulation shares similar vesicle size, drug encapsulation, surface charge. and toxicity profiles as liposomes formulated with cholesterol, however the tetradecanol preparation appeared to release considerably more drug during stability studies. Langmuir monolayer studies revealed that the condensing influence by tetradecanol is less than compared with cholesterol suggesting that this reduced intercalation by the former could explain why the tetradecanol formulation released more drug compared with cholesterol formulations. Environmental scanning electron microscopy (ESEM) was used to analyse the morphology and stability of liposomes. These investigations indicated that the presence of drugs within the liposomal bilayer were able to enhance the stability of the bilayers against collapse under reduced hydration conditions. In addition the presence of charged lipids within the formulation under reduced hydration conditions compared with its neutral counterpart. However the applicability of using ESEM as a new method to investigate liposome stability appears less valid than first hoped since the results are often open to varied interpretation and do not provide a robust set of data to support conclusions in some cases.

Application of SOA-NOLM in all-optical processing

This thesis presents experimental investigations of the use of semiconductor optical amplifiers in a nonlinear loop mirror (SOA-NOLM) and its application in all-optical processing. The techniques used are mainly experimental and are divided into three major applications. Initially the semiconductor optical amplifier, SOA, is experimentally characterised and the optimum operating condition is identified. An interferometric switch based on a Sagnac loop with the SOA as the nonlinear element is employed to realise all-optical switching. All-optical switching is a very attractive alternative to optoelectronic conversion because it avoids the conversion from the optical to the electronic domain and back again. The first major investigation involves a carrier suppressed return to zero, CSRZ, format conversion and transmission. This study is divided into single channel and four channel WDM respectively. The optical bandwidth which limits the conversion is investigated. The improvement of the nonlinear tolerance in the CSRZ transmission is shown which shows the suitability of this format for enhancing system performance. Second, a symmetrical switching window is studied in the SOA-NOLM where two similar control pulses are injected into the SOA from opposite directions. The switching window is symmetric when these two control pulses have the same power and arrive at the same time in the SOA. Finally, I study an all-optical circulating shift register with an inverter. The detailed behaviour of the blocks of zeros and ones has been analysed in terms of their transient measurement. Good agreement with a simple model of the shift register is obtained. The transient can be reduced but it will affect the extinction ratio of the pulses.

Symmetric operating point for increasing the extinction ratio of semiconductor based optical loop mirrors

Terahertz optical asymmetric demultiplexors (TOADs) use a semiconductor optical amplifier in an interferometer to create an all-optical switch and have potential uses in many optical networking applications. Here we demonstrate and compare experimentally a novel and simple method of dramatically increasing the extinction ratio of the device using a symmetrical configuration as compared to a ‘traditional’ configuration. The new configuration is designed to suppress the occurrence of self-switching in the device thus allowing signal pulses to be used at higher power levels. Using the proposed configuration an increase in extinction ratio of 10 dB has been measured on the transmitted port whilst benefiting from an improved input signal power handling capability.

The attenuation surface for contrast sensitivity has the form of a witch’s hat within the central visual field

Over the full visual field, contrast sensitivity is fairly well described by a linear decline in log sensitivity as a function of eccentricity (expressed in grating cycles). However, many psychophysical studies of spatial visual function concentrate on the central ±4.5 deg (or so) of the visual field. As the details of the variation in sensitivity have not been well documented in this region we did so for small patches of target contrast at several spatial frequencies (0.7–4 c/deg), meridians (horizontal, vertical, and oblique), orientations (horizontal, vertical, and oblique), and eccentricities (0–18 cycles). To reduce the potential effects of stimulus uncertainty, circular markers surrounded the targets. Our analysis shows that the decline in binocular log sensitivity within the central visual field is bilinear: The initial decline is steep, whereas the later decline is shallow and much closer to the classical results. The bilinear decline was approximately symmetrical in the horizontal meridian and declined most steeply in the superior visual field. Further analyses showed our results to be scale-invariant and that this property could not be predicted from cone densities. We used the results from the cardinal meridians to radially interpolate an attenuation surface with the shape of a witch's hat that provided good predictions for the results from the oblique meridians. The witch's hat provides a convenient starting point from which to build models of contrast sensitivity, including those designed to investigate signal summation and neuronal convergence of the image contrast signal. Finally, we provide Matlab code for constructing the witch's hat.

A series-dynamic-resistor-based converter protection scheme for doubly-fed induction generator during various fault conditions

This paper proposes a new converter protection method, primarily based on a series dynamic resistor (SDR) that avoids the doubly-fed induction generator (DFIG) control being disabled by crowbar protection during fault conditions. A combined converter protection scheme based on the proposed SDR and conventional crowbar is analyzed and discussed. The main protection advantages are due to the series topology when compared with crowbar and dc-chopper protection. Various fault overcurrent conditions (both symmetrical and asymmetrical) are analyzed and used to design the protection in detail, including the switching strategy and coordination with crowbar, and resistance value calculations. PSCAD/EMTDC simulation results show that the proposed method is advantageous for fault overcurrent protection, especially for asymmetrical faults, in which the traditional crowbar protection may malfunction.

Consensus on recording of gas permeable contact lens fit

Purpose: To develop a new schematic scheme for efficiently recording the key parameters of gas permeable contact lens (GP) fits based on current consensus. Methods: Over 100 established GP fitters and educators met to discuss the parameters proposed in educational material for evaluating GP fit and concluded on the key parameters that should be recorded. The accuracy and variability of evaluating the fluorescein pattern of GP fit was determined by having 35 experienced contact lens practitioners from across the world, grading 5 images of a range of fits and the topographer simulation of the same fits, in random, order using the proposed scheme. The accuracy of the grading was compared to objective image analysis of the fluorescein intensity of the same images. Results: The key information to record to adequately describe the fit of an GP was agreed as: the manufacturer, brand and lens parameters; settling time; comfort on a 5 point scale; centration; movement on blink on a ±2 scale; and the Primary Fluorescein Pattern in the central, mid-peripheral and edge regions of the lens averaged along the horizontal and vertical lens axes, on a ±2 scale. On average 50-60% of practitioners selected the median grade when subjectively rating fluorescein intensity and this was correlated to objective quantification (r= 0.602, p< 0.001). Objective grading suggesting horizontal median fluorescein intensity was generally symmetrical, as was the vertical meridian, but this was not the case for subjective grading. Simulated fluorescein patterns were subjectively and objectively graded as being less intense than real photographs (p< 0.01). Conclusion: GP fit recording can be standardised and simplified to enhance GP practice. © 2013 British Contact Lens Association.

Near-constant time delay response of fibre grating Sagnac loop filters

A theoretical and experimental investigation of the time delay characteristics of fiber Bragg grating-based Sagnac loops (FBGSLs) is presented. Analytic expressions for the phase and time delay of the FBGSL have been derived and excellent agreement is found between their predictions and experimental results for configurations incorporating uniform-period and chirped-period gratings. For symmetrical grating structures, it is found that the FBGSL time delay response is similar to that of the incorporated grating; with asymmetrical gratings, the FBGSL response is quite different. It is shown that wavelength-division-multiplexing filters exhibiting near-zero dispersion characteristics can be implemented using FBGSLs.

On the Mellin Transforms of Dirac’S Delta Function, The Hausdorff Dimension Function, and The Theorem by Mellin

Mathematics Subject Classification: 44A05, 46F12, 28A78

A fuzzy expected value approach under generalized data envelopment analysis

Fuzzy data envelopment analysis (DEA) models emerge as another class of DEA models to account for imprecise inputs and outputs for decision making units (DMUs). Although several approaches for solving fuzzy DEA models have been developed, there are some drawbacks, ranging from the inability to provide satisfactory discrimination power to simplistic numerical examples that handles only triangular fuzzy numbers or symmetrical fuzzy numbers. To address these drawbacks, this paper proposes using the concept of expected value in generalized DEA (GDEA) model. This allows the unification of three models - fuzzy expected CCR, fuzzy expected BCC, and fuzzy expected FDH models - and the ability of these models to handle both symmetrical and asymmetrical fuzzy numbers. We also explored the role of fuzzy GDEA model as a ranking method and compared it to existing super-efficiency evaluation models. Our proposed model is always feasible, while infeasibility problems remain in certain cases under existing super-efficiency models. In order to illustrate the performance of the proposed method, it is first tested using two established numerical examples and compared with the results obtained from alternative methods. A third example on energy dependency among 23 European Union (EU) member countries is further used to validate and describe the efficacy of our approach under asymmetric fuzzy numbers.

Approximate axial symmetries from continuous time quantum walks

The analysis of complex networks is usually based on key properties such as small-worldness and vertex degree distribution. The presence of symmetric motifs on the other hand has been related to redundancy and thus robustness of the networks. In this paper we propose a method for detecting approximate axial symmetries in networks. For each pair of nodes, we define a continuous-time quantum walk which is evolved through time. By measuring the probability that the quantum walker to visits each node of the network in this time frame, we are able to determine whether the two vertices are symmetrical with respect to any axis of the graph. Moreover, we show that we are able to successfully detect approximate axial symmetries too. We show the efficacy of our approach by analysing both synthetic and real-world data. © 2012 Springer-Verlag Berlin Heidelberg.

On some Sufficient Conditions for High Breakdown Point of ML Estimators

High breakdown point estimators LME(k) and LT E(k) for location and scale are obtained for symmetrical exponentially decreasing density family.