Optimization of random amplified polymorphic DNA techniques for use in genetic studies of Cuban triatominae

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4) generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA). Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.

Vision-Based Assisted Teleoperation for Inspection Tasks with a Small ROV

It is well-known that ROVs require human intervention to guarantee the success of their assignment, as well as the equipment safety. However, as its teleoperation is quite complex to perform, there is a need for assisted teleoperation. This study aims to take on this challenge by developing vision-based assisted teleoperation maneuvers, since a standard camera is present in any ROV. The proposed approach is a visual servoing solution, that allows the user to select between several standard image processing methods and is applied to a 3-DOF ROV. The most interesting characteristic of the presented system is the exclusive use of the camera data to improve the teleoperation of an underactuated ROV. It is demonstrated through the comparison and evaluation of standard implementations of different vision methods and the execution of simple maneuvers to acquire experimental results, that the teleoperation of a small ROV can be drastically improved without the need to install additional sensors.

Bioelectrochemically-assisted reductive dechlorination of 1,2-dichloroethane by a Dehalococcoides-enriched microbial culture

The aim of this study was to verify the possibility to use a polarized graphite electrode as an electron donor for the reductive dechlorination of 1,2-dichloroethane, an ubiquitous groundwater contaminant. The rate of 1,2-DCA dechlorination almost linearly increased by decreasing the set cathode potential over a broad range of set cathode potentials (i.e., from −300 mV to −900 mV vs. the standard hydrogen electrode). This process was primarily dependent on electrolytic H2 generation. On the other hand, reductive dechlorination proceeded (although quite slowly) with a very high Coulombic efficiency (near 70%) at a set cathode potential of −300 mV, where no H2 production occurred. Under this condition, reductive dechlorination was likely driven by direct electron uptake from the surface of the polarized electrode. Taken as a whole, this study further extends the range of chlorinated contaminants which can be treated with bioelectrochemical systems.

A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques

The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.

Comparison of direct immunofluorescence, conventional cell culture and polymerase chain reaction techniques for detecting respiratory syncytial virus in nasopharyngeal aspirates from infants

A total of 316 samples of nasopharyngeal aspirate from infants up to two years of age with acute respiratory-tract illnesses were processed for detection of respiratory syncytial virus (RSV) using three different techniques: viral isolation, direct immunofluorescence, and PCR. Of the samples, 36 (11.4%) were positive for RSV, considering the three techniques. PCR was the most sensitive technique, providing positive findings in 35/316 (11.1%) of the samples, followed by direct immunofluorescence (25/316, 7.9%) and viral isolation (20/315, 6.3%) (p < 0.001). A sample was positive by immunofluorescence and negative by PCR, and 11 (31.4%) were positive only by RT-PCR. We conclude that RT-PCR is more sensitive than IF and viral isolation to detect RSV in nasopharyngeal aspirate specimens in newborn and infants.

Adhesive selection for single lap bonded joints: Experimentation and advanced techniques for strength prediction

The integrity of multi-component structures is usually determined by their unions. Adhesive-bonding is often used over traditional methods because of the reduction of stress concentrations, reduced weight penalty, and easy manufacturing. Commercial adhesives range from strong and brittle (e.g., Araldite® AV138) to less strong and ductile (e.g., Araldite® 2015). A new family of polyurethane adhesives combines high strength and ductility (e.g., Sikaforce® 7888). In this work, the performance of the three above-mentioned adhesives was tested in single lap joints with varying values of overlap length (LO). The experimental work carried out is accompanied by a detailed numerical analysis by finite elements, either based on cohesive zone models (CZM) or the extended finite element method (XFEM). This procedure enabled detailing the performance of these predictive techniques applied to bonded joints. Moreover, it was possible to evaluate which family of adhesives is more suited for each joint geometry. CZM revealed to be highly accurate, except for largely ductile adhesives, although this could be circumvented with a different cohesive law. XFEM is not the most suited technique for mixed-mode damage growth, but a rough prediction was achieved.

Adhesive thickness effects of a ductile adhesive by optical measurement techniques

Adhesive bonding is an excellent alternative to traditional joining techniques such as welding, mechanical fastening or riveting. However, there are many factors that have to be accounted for during joint design to accurately predict the joint strength. One of these is the adhesive layer thickness (tA). Most of the results are for epoxy structural adhesives, tailored to perform best with small values of tA, and these show that the lap joint strength decreases with increase of tA (the optimum joint strength is usually obtained with tA values between 0.1 and 0.2 mm). Recently, polyurethane adhesives were made available in the market, designed to perform with larger tA values, and whose fracture behaviour is still not studied. In this work, the effect of tA on the tensile fracture toughness (View the MathML source) of a bonded joint is studied, considering a novel high strength and ductile polyurethane adhesive for the automotive industry. This work consists on the fracture characterization of the bond by a conventional and the J-integral techniques, which accurately account for root rotation effects. An optical measurement method is used for the evaluation of crack tip opening (δn) and adherends rotation at the crack tip (θo) during the test, supported by a Matlab® sub-routine for the automated extraction of these parameters. As output of this work, fracture data is provided in traction for the selected adhesive, enabling the subsequent strength prediction of bonded joints.

Tensile fracture characterization of adhesive joints by standard and optical techniques

The use of adhesive joints has increased in recent decades due to its competitive features compared with traditional methods. This work aims to estimate the tensile critical strain energy release rate (GIC) of adhesive joints by the Double-Cantilever Beam (DCB) test. The J-integral is used since it enables obtaining the tensile Cohesive Zone Model (CZM) law. An optical measuring method was developed for assessing the crack tip opening (δn) and adherends rotation (θo). The proposed CZM laws were best approximated by a triangular shape for the brittle adhesive and a trapezoidal shape for the two ductile adhesives.

Biosynthesis-Assisted Structural Elucidation of the Bartolosides, Chlorinated Aromatic Glycolipids from Cyanobacteria

The isolation of the bartolosides, unprecedented cyanobacterial glycolipids featuring aliphatic chains with chlorine substituents and C-glycosyl moieties, is reported. Their chlorinated dialkylresorcinol (DAR) core presented a major structural-elucidation challenge. To overcome this, we discovered the bartoloside (brt) biosynthetic gene cluster and linked it to the natural products through in vitro characterization of the DAR-forming ketosynthase and aromatase. Bioinformatic analysis also revealed a novel potential halogenase. Knowledge of the bartoloside biosynthesis constrained the DAR core structure by defining key pathway intermediates, ultimately allowing us to determine the full structures of the bartolosides. This work illustrates the power of genomics to enable the use of biosynthetic information for structure elucidation.

Effects of self-administered exercises based on Tuina techniques on musculoskeletal disorders of professional orchestra musicians: a randomized controlled trial

Background Musicians are frequently affected by playing-related musculoskeletal disorders (PRMD). Common solutions used by Western medicine to treat musculoskeletal pain include rehabilitation programs and drugs, but their results are sometimes disappointing. Objective To study the effects of self-administered exercises based on Tuina techniques on the pain intensity caused by PRMD of professional orchestra musicians, using numeric visual scale (NVS). Design, setting, participants and interventions We performed a prospective, controlled, single-blinded, randomized study with musicians suffering from PRMD. Participating musicians were randomly distributed into the experimental (n = 39) and the control (n = 30) groups. After an individual diagnostic assessment, specific Tuina self-administered exercises were developed and taught to the participants. Musicians were instructed to repeat the exercises every day for 3 weeks. Main outcome measures Pain intensity was measured by NVS before the intervention and after 1, 3, 5, 10, 15 and 20 d of treatment. The procedure was the same for the control group, however the Tuina exercises were executed in points away from the commonly-used acupuncture points. Results In the treatment group, but not the control group, pain intensity was significantly reduced on days 1, 3, 5, 10, 15 and 20. Conclusion The results obtained are consistent with the hypothesis that self-administered exercises based on Tuina techniques could help professional musicians controlling the pain caused by PRMD. Although our results are very promising, further studies are needed employing a larger sample size and double blinding designs.

Immediate effects of Tuina techniques on working-related musculoskeletal disorder of professional orchestra musicians

BACKGROUND: Musicians are a prone group to suffer from working-related musculoskeletal disorder (WRMD). Conventional solutions to control musculoskeletal pain include pharmacological treatment and rehabilitation programs but their efficiency is sometimes disappointing. OBJECTIVE: The aim of this research is to study the immediate effects of Tuina techniques on WRMD of professional orchestra musicians from the north of Portugal. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: We performed a prospective, controlled, single-blinded, randomized study. Professional orchestra musicians with a diagnosis of WRMD were randomly distributed into the experimental group (n=39) and the control group (n=30). During an individual interview, Chinese diagnosis took place and treatment points were chosen. Real acupoints were treated by Tuina techniques into the experimental group and non-specific skin points were treated into the control group. Pain was measured by verbal numerical scale before and immediately after intervention. RESULTS: After one treatment session, pain was reduced in 91.8% of the cases for the experimental group and 7.9% for the control group. CONCLUSION: Although results showed that Tuina techniques are effectively reducing WRMD in professional orchestra musicians of the north of Portugal, further investigations with stronger measurements, double-blinding designs and bigger simple sizes are needed.

Efficiency of strengthening techniques assessed for existing masonry buildings

The Azores archipelago is a zone with a vast cultural heritage, presenting a building stock mainly constructed in traditional stone masonry. It is known that this type of construction exhibits poor behaviour under seismic excitations; however it is extensively used in seismic prone areas, such as this case. The 9th of July of 1998 earthquake was the last seismic event in the islands, leaving many traditional stone constructions severely damaged or totally destroyed. This scenario led to an effort by the local government of improving the seismic resistance of these constructions, with the application of several reinforcement techniques. This work aims to study some of the most used reinforcement schemes after the 1998 earthquake, and to assess their effectiveness in the mitigation of the construction’s seismic vulnerability. A brief evaluation of the cost versus benefit of these retrofitting techniques is also made, seeking to identify those that are most suitable for each building typology. Thus, it was sought to analyze the case of real structures with different geometrical and physical characteristics, by establishing a comparison between the seismic performance of reinforced and non-reinforced structures. The first section contains the analysis of a total of six reinforcement scenarios for each building chosen. Using the recorded 1998 earthquake accelerograms, a linear time-history analysis was performed for each reinforcement scenario. A comparison was then established between the maximum displacements, inter-storey drift and maximum stress obtained, in order to evaluate the global seismic response of each reinforced structure. In the second part of the work, the examination of the performance obtained in the previous section, in relation to the cost of implementing each reinforcement technique, allowed to draw conclusions concerning the viability of implementing each reinforcement method, based on the book value of the buildings in study.

Algorithms for information extraction and signal annotation on long-term biosignals using clustering techniques

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

Urban land cover change detection analysis and modeling spatio-temporal Growth dynamics using Remote Sensing and GIS Techniques: A case study of Dhaka, Bangladesh

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

Investigating the use of dasymetric techniques for assessing employment containment in Melbourne, Australia

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.