Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.

A systematic procedure to enhance reproducibility of SWASV cycles in the determination of toxic metals in real samples

This work presents for the first time a systematic study on the optimization of the electrochemical cleaning time of a mercury film when it is used as a working electrode material in the analysis of toxic metals, such as Pb2+, used as model metal, in real samples by SWASV. The optimization study for the film’s cleaning time aimed at attaining a Pb2+ minimum value in the film after the re-oxidation step of the pre-concentrated metal, given the impossibility of complete removal of traces of the electroactive species from the film. This value was kept constant in each concentration range studied ensuring thus that all assays were performed in initial identical conditions. An assay performed on a synthetic sample was taken as reference. In it, given the absence of matrix effects, and after the electrochemical cleaning step, a direct proportionality was observed between the residual amounts of Pb2+ in the film (which for the cleaning time used was never completely removed) and Pb2+ concentration in the solution. This fact determined a high correlation between Pb2+ peak current and Pb2+ concentration which was not observed when real samples (tree leaves) were analyzed. This behavior may result from the presence of the interfering surfactants always present in real samples of complex matrix. Cleaning time optimization was performed for the following Pb2+ concentration ranges in the real samples of complex matrix: 0.006-0.020, 0.020-0.080, 0.060-0.200 and 0.100-0.600 ppb. As expected, in order to obtain identical levels of film’s cleaning efficiency, the need for longer cleaning times has been observed for higher concentrations. The optimized cleaning times for the concentration ranges under study were 120, 150, 180 e 300 s, respectively.

Surgical correction of scoliosis: Numerical analysis and optimization of the procedure

A previously developed model is used to numerically simulate real clinical cases of the surgical correction of scoliosis. This model consists of one-dimensional finite elements with spatial deformation in which (i) the column is represented by its axis; (ii) the vertebrae are assumed to be rigid; and (iii) the deformability of the column is concentrated in springs that connect the successive rigid elements. The metallic rods used for the surgical correction are modeled by beam elements with linear elastic behavior. To obtain the forces at the connections between the metallic rods and the vertebrae geometrically, non-linear finite element analyses are performed. The tightening sequence determines the magnitude of the forces applied to the patient column, and it is desirable to keep those forces as small as possible. In this study, a Genetic Algorithm optimization is applied to this model in order to determine the sequence that minimizes the corrective forces applied during the surgery. This amounts to find the optimal permutation of integers 1, ... , n, n being the number of vertebrae involved. As such, we are faced with a combinatorial optimization problem isomorph to the Traveling Salesman Problem. The fitness evaluation requires one computing intensive Finite Element Analysis per candidate solution and, thus, a parallel implementation of the Genetic Algorithm is developed.

Claustrophobia and adherence in magnetic resonance imaging procedure

Claustrophobia causes a huge discomfort to those who need to perform Magnetic Resonance examinations mainly due to the physical design of most equipment. This study aimed to maximize the success rate of Magnetic Resonance Imaging (MRI) clinical studies in claustrophobic patients by the identification of facilitative strategies.

Fast screening procedure for antibiotics in wastewaters by direct HPLC-DAD analysis

Growing concern about the contamination of wastewaters by antibiotics demands fast but sensitive analytical methodologies, for the screening of a large number of samples. The purpose of this work was to develop a simple methodology, using direct injection of the samples, by HPLC with diode array detection (DAD), for a multiresidue analysis of five antibiotics of different classes. Wastewater from an urban water treatment plant was selected as a model to study possible coelution of interfering compounds. The linearity interval ranged from 40 to 400 µg/L for amoxicillin (Amox), metronidazole (Metro), cefazolin (Cefa), and chloramphenicol (Chloram) and from 20 to 200 µg/L for sulfamethoxazole (Sulfa), with LODs lower than 14 µg/L. Repeatability, expressed by the CV of six repeated injections, ranged from 1 to 8%, while the intermediate precision varied between 2 and 11%. The recovery ranged from 90 to 109%. This method enables the fast screening of a large number of samples, with an expanded uncertainty in the 1–22% range. The advantage of the proposed method is to significantly reduce the number of samples to be analyzed by more complex methods.

Optimization of QuEChERS procedure coupled to GC-ECD for organochlorine pesticide determination in carrot samples

An optimised version of the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method for simultaneous determination of 14 organochlorine pesticides in carrots was developed using gas chromatography coupled with electron-capture detector (GC-ECD) and confirmation by gas chromatography tandem mass spectrometry (GC-MS/MS). A citrate-buffered version of QuEChERS was applied for the extraction of the organochlorine pesticides, and for the extract clean-up, primary secondary amine, octadecyl-bonded silica (C18), magnesium sulphate (MgSO4) and graphitized carbon black were used as sorbents. The GC-ECD determination of the target compounds was achieved in less than 20 min. The limits of detection were below the EUmaximum residue limits (MRLs) for carrots, 10–50 μg kg−1, while the limit of quantification did exceed 10 μg kg−1 for hexachlorobenzene (HCB). The introduction of a sonication step was shown to improve the recoveries. The overall average recoveries in carrots, at the four tested levels (60, 80, 100 and 140 μg kg−1), ranged from 66 to 111% with relative standard deviations in the range of 2– 15 % (n03) for all analytes, with the exception of HCB. The method has been applied to the analysis of 21 carrot samples from different Portuguese regions, and β-HCH was the pesticide most frequently found, with concentrations oscillating between less than the limit of quantification to 14.6 μg kg−1. Only one sample had a pesticide residue (β-HCH) above the MRL, 14.6 μg kg−1. This methodology combines the advantages of both QuEChERS and GC-ECD, producing a very rapid, sensitive and reliable procedure which can be applied in routine analytical laboratories.

Validation of a single-extraction procedure for sequential analysis of vitamin E, cholesterol, fatty acids, and total fat in seafood

Food lipid major components are usually analyzed by individual methodologies using diverse extractive procedures for each class. A simple and fast extractive procedure was devised for the sequential analysis of vitamin E, cholesterol, fatty acids, and total fat estimation in seafood, reducing analyses time and organic solvent consumption. Several liquid/liquid-based extractive methodologies using chlorinated and non-chlorinated organic solvents were tested. The extract obtained is used for vitamin E quantification (normal-phase HPLC with fluorescence detection), total cholesterol (normal-phase HPLC with UV detection), fatty acid profile, and total fat estimation (GC-FID), all accomplished in <40 min. The final methodology presents an adequate linearity range and sensitivity for tocopherol and cholesterol, with intra- and inter-day precisions (RSD) from 3 to 11 % for all the components. The developed methodology was applied to diverse seafood samples with positive outcomes, making it a very attractive technique for routine analyses in standard equipped laboratories in the food quality control field.

Timing analysis of an inter-cell mobility procedure for a wired/wireless PROFIBUS network

Recently, there have been a few research efforts towards extending the capabilities of fieldbus networks to encompass wireless support. In previous works we have proposed a hybrid wired/wireless PROFIBUS network solution where the interconnection between the heterogeneous communication media was accomplished through bridge-like interconnecting devices. The resulting networking architecture embraced a Multiple Logical Ring (MLR) approach, thus with multiple independent tokens, where the communication between different domains was supported by the Inter-Domain Protocol (IDP). The proposed architecture also supports mobility of stations between different wireless cells. To that hybrid wired/wireless networking architecture we have proposed a worst-case response timing analysis of the IDP, without considering inter-cell mobility (or handoff) of stations. In this paper, we advance that previous work by proposing a worst-case timing analysis of the mobility procedure.

Poster abstract: Smart-HOP: a reliable handoff procedure for supporting mobility in wireless sensor networks

This poster abstract presents smart-HOP, a reliable handoff mechanism for mobility support in Wireless Sensor Networks (WSNs). This technique relies on a fuzzy logic approach applied at two levels: the link quality estimation level and the access point selection level. We present the conceptual design of smart-HOP and then we discuss implementation requirements and challenges.

On a reliable handoff procedure for supporting mobility in wireless sensor networks

Wireless sensor network (WSN) applications such as patients’ health monitoring in hospitals, location-aware ambient intelligence, industrial monitoring /maintenance or homeland security require the support of mobile nodes or node groups. In many of these applications, the lack of network connectivity is not admissible or should at least be time bounded, i.e. mobile nodes cannot be disconnected from the rest of the WSN for an undefined period of time. In this context, we aim at reliable and real-time mobility support in WSNs, for which appropriate handoff and rerouting decisions are mandatory. This paper1 drafts a mechanism and correspondent heuristics for taking reliable handoff decisions in WSNs. Fuzzy logic is used to incorporate the inherent imprecision and uncertainty of the physical quantities at stake.

Experimental measuring procedure for the friction torque in rolling bearings

The reduction of the power loss generated in mechanical transmissions and the use of low friction biodegradable lubricants has been attracting considerable attention in recent times. Therefore, it is necessary to develop methods to test and evaluate the performance of such lubricants and compare them with conventional ones. In this sense, a Four-Ball Machine was modified allowing the test of rolling bearings. A 51107 thrust ball bearing was used to test two different greases and the corresponding base oils. Friction torque and operating temperatures were continuously monitored to quantify the power loss and the heat evacuation for each lubricant tested. Copyright © 2010 John Wiley & Sons, Ltd.

Development of a meso-microscale coupling procedure for site assessment in complex terrain

A procedure for coupling mesoscale and CFD codes is presented, enabling the inclusion of realistic stratification flow regimes and boundary conditions in CFD simulations of relevance to site and resource assessment studies in complex terrain. Two distinct techniques are derived: (i) in the first one, boundary conditions are extracted from mesoscale results to produce time-varying CFD solutions; (ii) in the second case, a statistical treatment of mesoscale data leads to steady-state flow boundary conditions believed to be more representative than the idealised profiles which are current industry practice. Results are compared with measured data and traditional CFD approaches.

Automated synthesis procedure of RF discrete tuning differential capacitance circuits

The paper presents a RFDSCA automated synthesis procedure. This algorithm determines several RFDSCA circuits from the top-level system specifications all with the same maximum performance. The genetic synthesis tool optimizes a fitness function proportional to the RFDSCA quality factor and uses the epsiv-concept and maximin sorting scheme to achieve a set of solutions well distributed along a non-dominated front. To confirm the results of the algorithm, three RFDSCAs were simulated in SpectreRF and one of them was implemented and tested. The design used a 0.25 mum BiCMOS process. All the results (synthesized, simulated and measured) are very close, which indicate that the genetic synthesis method is a very useful tool to design optimum performance RFDSCAs.

On sampling collection procedure effectiveness for forest soil characterization

One of the most important measures to prevent wild forest fires is the use of prescribed and controlled burning actions as it reduce the fuel mass availability. The impact of these management activities on soil physical and chemical properties varies according to the type of both soil and vegetation. Decisions in forest management plans are often based on the results obtained from soil-monitoring campaigns. Those campaigns are often man-labor intensive and expensive. In this paper we have successfully used the multivariate statistical technique Robust Principal Analysis Compounds (ROBPCA) to investigate on the sampling procedure effectiveness for two different methodologies, in order to reflect on the possibility of simplifying and reduce the sampling collection process and its auxiliary laboratory analysis work towards a cost-effective and competent forest soil characterization.

Elution of proteins from polyacrylamide eels: a simple and economic procedure

A simplified methodology for the quantitative electroelution of proteins from polyacrylamide gels is described. After staining with Coomassie Brilliant Blue R 250, the identified bands are excised from the gel and the proteins eluted using a procedure developed for use in conventional tube gel electrophoresis equipment.