Electrochemical genosensor based on colloidal gold nanoparticles for the detection of Factor V Leiden mutation using disposable pencil graphite electrodes

Electrochemical genosensors for the detection of the Factor V Leiden mutation from polymerase chain reaction (PCR) amplicons using the oxidation signal of colloidal gold (Au) is described. A pencil graphite electrode (PGE) modified with target DNA, when hybridized with complementary probes conjugated to Au nanoparticles, responded with the appearance of a Au oxide wave at approximately +1.20 V. Specific probes were immobilized onto the Au nanoparticles in two different modes: (a) Inosine-substituted probes were covalently attached from their amino groups at the 5' end using N-(3-dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent onto a carboxylate-terminated l-cysteine self-assembled monolayer (SAM) preformed on the Au nanoparticles, and (b) probes with a hexanethiol group at their 5' phosphate end formed a SAM on Au nanoparticles. The genosensor relies on the hybridization of the probes with their complementary targets, which are covalently immobilized at the PGE surface. Au-tagged 23-mer capture probes were challenged with the synthetic 23-mer target, 131-base single-stranded DNA or denatured 256-base polymerase chain reaction (PCR) amplicon. The appearance of the Au oxidation signal shortened the assay time and simplified the detection of the Factor V Leiden mutation from PCR amplified real samples. The discrimination between the homozygous and heterozygous mutations was also established by comparing the peak currents of the Au signals. Numerous factors affecting the hybridization and nonspecific binding events were optimized. The detection limit for the PCR amplicons was found to be as low as 0.78 fmol; thus, it is suitable for point-of-care applications.     

An evolutionary algorithm-based decision support system for managing flexible manufacturing

The high investment cost of flexible manufacturing systems (FMS) requires their management to be effective and efficient. The effectiveness in managing FMSs includes addressing machine loading, scheduling parts and dispatching vehicles and the quality of the solution. Therefore the problem is inevitably multi-criteria, and decision maker's judgement may contribute to the quality of the solution and the systems's performance. On the other hand, each of these problems of FMS is hard to optimize due to the large and discrete solution spaces (NP-hard). The FMS manager must address each of these problems hierarchically (separately) or simultaneously (aggregately) in a limited time. The efficiency of the management is related to the response time.

Here we propose a decision support system that utilizes an evolutionary algorithm (EA) with a memory of “good” past experiments as the solution engine. Therefore, even in the absence of an expert decision maker the performance of the solution engine and/or the quality of the solutions are maintained.

The experiences of the decision maker(s) are collected in a database (i.e., memory-base) that contains problem characteristics, the modeling parameters of the evolutionary program, and the quality of the solution. The solution engine in the decision support system utilizes the information contained in the memory-base in solving the current problem. The initial population is created based on a memory-based seeding algorithm that incorporates information extracted from the quality solutions available in the database. Therefore, the performance of the engine is designed to improve following each use gradually. The comparisons obtained over a set of randomly generated test problems indicate that EAs with the proposed memory-based seeding perform well. Consequently, the proposed DSS improves not only the effectiveness (better solution) but also the efficiency (shorter response time) of the decision maker(s).     

Frequency and voltage dependence of dielectric properties,complex electric modulus,and electrical conductivity in Au/7% graphene doped‐PVA/n‐Si (MPS) structures

In order to increase the capacitance of Au/n‐Si (MS) structure, 7% graphene doped PVA was coated on n‐Si as an interfacial layer. The measured data of capacitance (C) and conductance (G/ω) of Au/7% graphene doped‐PVA/n‐Si (MPS) structure was utilized for the calculation of real and imaginary parts of complex permittivity (ε^* = ε′ − jε″), loss tangent (tanδ), complex electric modulus (M^* = M′ + jM″), and electrical conductivity (σ). The admittance measurements (C and G/ω) were carried out in the frequency range of 0.5 kHz to 1 MHz at room temperature. Frequency dependence of the dielectric constant (ε′), dielectric loss (ε″) and tanδ shows a dispersive behavior at low frequencies. This behavior was explained by Maxwell–Wagner relaxation. Due to the dipolar and the interfacial polarizations, as well as the surface states (N_ss) and the interfacial PVA layer, the parameters exhibited a strong dependence on frequency and applied bias voltage. The σ versus log(f) plot exhibited both low and high frequency dispersion phenomena such that at low frequencies σ value corresponding to the dc conductivity (σ_dc), but at high frequencies it corresponds to the ac conductivity (σ_ac). M′ and M″, both, have low values in the low frequency region. However, an increase is observed with the increasing frequency due to the short‐range mobility of charge carriers. As a result, the change in dielectric parameters and electric modulus with frequency is the result of relaxation phenomena and surface states. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43827.     

Mucin profiles of the abomasum in bulls and rams: A comparative study

Many pathogens require direct binding to mucosal cells to cause an infection. The mucosal epithelium of the digestive tract, which is covered by a mucin layer, fulfills several protective functions that are essential to maintaining the health of the digestive tract. Mucins are glycoproteins, which are found on membranes and in mucus gels and protect the underlying mucosal cells. Both membrane‐associated mucins and secreted mucins are critical components of mucosal defense. The aim of this study was to determine the localization and expression of mucin profile of the abomasum via histochemistry and immunohistochemistry. The abomasums of 20 bulls and 20 rams were evaluated. Histochemical examination showed that neutral and acidic mucins were present in the mucosa and the glands of the pars cardiaca, fundus, and pars pylorica of the abomasums of both bulls and rams. However, the expression of acidic mucins was weak in the superficial glands and strong in the deep glands of the abomasum of rams. In both bulls and rams, MUC1, MUC5AC, and MUC6 were expressed in the glandular epithelial cells in all regions of the abomasum. Interestingly, while MUC2 was not expressed in the pars cardiaca and fundus, it was weakly expressed in the parietal cells of the pars pylorica in both species. In conclusion, the presence of neutral and acidic mucins and MUC1, MUC2, MUC5AC, and MUC6 proteins in luminal epithelial and glandular cells of abomasum in the bulls and rams support the hypothesis that mucins play a key role in the protection of the abomasal mucosa against infectious agents.     

Determination of Rosmarinic Acid by High-Performance Liquid Chromatography and Its Application to Certain Salvia Species and Rosemary

This study describes a method development for the determination of rosmarinic acid (RA) by using a gradient high-performance liquid chromatography (HPLC) and its application to certain plant materials. The analysis was performed by utilizing a two solvents system [A: methanol/water/formic acid (10:88:2; v:v:v); B: methanol/water/formic acid (90:8:2; v:v:v)] on a reverse-phase column. The flow rate and injection volume were 1 ml min⁻¹ and 10 μl, respectively. Signals were detected at 280 nm. In addition, an internal standard (IS) technique was applied for the analysis of RA to increase precision, and propylparaben was employed for this purpose. The repeatability results as RSD% were 1.66, 1.17 and 1.26 for intra-day and 1.38 was for inter-day with the employment of (3.67 × 10⁻⁵ M) RA. A limit of linearity (LOL) was observed in a wide (1.13 × 10⁻⁵–5.65 × 10⁻⁴ M) concentration range. Linearity parameters were also examined in the range of 5.95 × 10⁻⁶–7.14 × 10⁻⁵ M RA, and very good correlation was observed. The limit of detection (LOD) and limit of quantification (LOQ) (for inter-day) were 1.60 × 10⁻⁶ M (signal/noise [S/N] = 3.3) and 4.80 × 10⁻⁶ M (S/N = 10), respectively. The method was applied to the extracts of certain Lamiaceae plants (Salvia candidissima Vahl. subsp. candidissima, S. sclarea L., S. verticillata L. subsp. verticillata and R. officinalis L.), and reasonable results were obtained.     

Development of an intelligent motion controller and its application to the automation of a McBain-Bakr balance

This paper presents a modular approach to motion control using a microconlroller-based stepper motor driver. As an application of the driver, an inexpensive and portable one-dimensional robot was built to automate an existing experimental setup for measurement of adsorption isotherms using the McBain-Bakr technique. Automatic recording of position versus time yields data for the sludy of adsorption/desorption dynamics.     

Statistical fibre failure and single crack behaviour in uniaxially reinforced ceramic composites

A lower bound to the work of pull-out is estimated for a ceramic composite under uniaxial loading assuming that matrix crumbling does not occur. Fibre failure is assumed to be governed by Weibull distribution. In order to compute a lower bound to the energy dissipation it is assumed that the failure occurs by a single matrix crack. The fibre/matrix interface is assumed to be constrained by friction only. The work of pull-out estimated from the present model is compared to the energy dissipation before fibre failure which was computed by Aveston, Cooper and Kelly (ACK) in 1971. Comparisons are made with the surface energy of monolithic materials.     

A pattern-based genetic algorithm for multi-robot coverage path planning minimizing completion time

Sensor-based multi-robot coverage path planning problem is one of the challenging problems in managing flexible, computer-integrated, intelligent manufacturing systems. A novel pattern-based genetic algorithm is proposed for this problem. The area subject to coverage is modeled with disks representing the range of sensing devices. Then the problem is defined as finding a sequence of the disks for each robot to minimize the coverage completion time determined by the maximum time traveled by a robot in a mobile robot group. So the environment needs to be partitioned among robots considering their travel times. Robot turns cause the robot to slow down, turn and accelerate inevitably. Therefore, the actual travel time of a mobile robot is calculated based on the traveled distance and the number of turns. The algorithm is designed to handle routing and partitioning concurrently. Experiments are conducted using P3-DX mobile robots in the laboratory and simulation environment to validate the results.