A DNA methylation assay for detection of ovarian cancer cells using a HpaII/MspI digestion-based PCR assay in an integrated microfluidic system

Early and accurate diagnosis of cancer plays a very important role in favorable clinical outcomes. DNA methylation of tumor suppressor genes has been recognized as a diagnostic biomarker for early carcinogenesis. The presence of 5-methylcytosine in the CpG islands in the promoter region of a tumor suppressor gene is an important indicator of DNA methylation. However, the standard detection assay utilizing a bisulfite treatment and HpaII/MspI endonuclease digestion is a tedious and lengthy process and requires a relatively large amount of DNA for testing. In this study, the methylated DNAs of various tumor suppressor genes, HAAO, HOXA9 and SFRP5, were chosen as candidates for detection of ovarian cancer cells. The entire experimental process for the DNA methylation assay, including target DNA isolation, HpaII/MspI endonuclease digestion, and nucleic acid amplification has been realized in an integrated microfluidic system. The limit of detection using this developed system has been experimentally determined to be 10² cells/reaction. The entire process from sample loading to analysis of the results only took 3 h which is much faster than the existing protocols. Different sources of biosamples, such as cells, ascites and serums, could be detected with the methylated DNA, indicating that this developed microfluidic system could be adapted for clinical use. Thus, this developed microsystem may be a promising platform for the rapid and early diagnosis of cancers.     

Identification of respiratory pathogen Bordetella Pertussis using integrated microfluidic chip technology

Integrated PCR–CE chip technology has immense potential to be applied in clinical diagnostics. In this work we demonstrate the application of our integrated PCR–CE chip for the detection of the respiratory pathogen Bordetella pertussis. A series of experiments with varying cell concentrations (200,000–2 cfu) were performed to obtain the analytical detection limits of the chip. We find that the chip technology is well suited for sensitive detection of Bordetella pertussis, using genetic material from less than even 2 cfu. We also utilized an off-chip real-time PCR method to compare and validate our on-chip approach.     

An integrated microfluidic system using mannose-binding lectin for bacteria isolation and biofilm-related gene detection

Molecular diagnosis of biofilm-related genes (BRGs) in common bacteria that cause periprosthetic joint infections may provide crucial information for clinicians. In this study, several BRGs, including ica, fnbA, and fnbB, were rapidly detected (within 1 h) with a new integrated microfluidic system. Mannose-binding lectin (MBL)-coated magnetic beads were used to isolate these bacteria, and on-chip nucleic acid amplification (polymerase chain reaction, PCR) was then performed to detect BRGs. Both eukaryotic and prokaryotic MBLs were able to isolate common bacterial strains, regardless of their antibiotic resistance, and limits of detection were as low as 3 and 9 CFU for methicillin-resistant Staphylococcus aureus and Escherichia coli, respectively, when using a universal 16S rRNA PCR assay for bacterial identification. It is worth noting that the entire process including bacteria isolation by using MBL-coated beads for sample pre-treatment, on-chip PCR, and fluorescent signal detection could be completed on an integrated microfluidic system within 1 h. This is the first time that an integrated microfluidic system capable of detecting BRGs by using MBL as a universal capturing probe was reported. This integrated microfluidic system might therefore prove useful for monitoring profiles of BRGs and give clinicians more clues for their clinical judgments in the near future.     

Implementation of a decision support system using an interactive large-scale high-resolution display

In this research, we propose and evaluate a decision support system using an interactive large-scale high-resolution display. This decision support system supports the summarization and decision-making of a large amount of disaster information during the occurrence of a large-scale natural disaster. Municipal employees at the disaster control headquarters can display disaster information on the large-scale display with a touch or flick on a laptop or tablet. To evaluate the operability, readability, functionality, and necessity of the decision support system, we surveyed 23 municipal employees in the disaster prevention division using a questionnaire. The system received a great evaluation in all the evaluation items.     

Isolation of magnetically tagged cancer cells through an integrated magnetofluidic device

Circulating tumour cells (CTC) in the bloodstream has been implicated in cancer metastasis. Efficient removal of CTC could potentially be an effective therapeutic measure against cancer metastasis. In this study, the hydrodynamic focusing flow in microfluidic channels (R _e  ? 1) was considered together with the magnetophoretic force. The localised magnetic field was achieved through a passivated current-carrying multilayered microstripline, where the generated field gradient was used to attract the magnetic beads to the desired outlet. The experimental results show that the device is capable of isolating purely magnetic beads with an efficiency of 91 % while isolation efficiency of the magnetically tagged HeLa cervical cancer cells from cell suspension yielded an isolation efficiency of 79 %.     

The application of an expert system shell to an unstructured domain of expertise: using expert system technology to teach SSM

Action research is orchestrated by the interaction between theory and practice as exemplified by soft systems methodology (SSM). The use of computers has traditionally been developed out of what has been referred to as the ‘rationalistic’ tradition which represents the search for solutions for pre-determined, logical goals (Winograd & Flores, 1986). Action research anticipates the need to explore and learn about each intervention into human activity, using models thought to be relevant to thinking about the world to help structure the learning process. In comparison, the ‘rationalistic’ tradition seeks universal laws for behaviour and uses validated models of the reality represented by these laws to solve problems. It would seem, therefore, that to translate SSM into a computer-based format would be dialectically opposed to the intellectual framework in which the methodology is seated. However, the authors suggest that there is one area in which a computer-based version of SSM may be useful. This paper describes an exploratory study of the application of expert system technology as an aid to teaching SSM.     

Effects of transition to an integrated IT technology in surveying work

An integrated IT technology, characterized by a change in information support from a strong specialization towards greater integration has been introduced within the surveying company in Sweden. The aim of this study is to compare and describe effects of the transition to this new information technology in relation to job and organizational characteristics and effectiveness and well-being measures between 1998 and 2000. The results show that a positive attitude to the new IT system increased from 1998 to 2000. The transition to a new IT technology had negative consequences on job content and job control in 1999 but improvements could be seen in 2000. Co-operation with clients and service quality to clients improved each year from 1998 to 2000. Generally, a positive attitude to IT integration was related to high continuous improvement practices, goal clarity and job control in all three phases. However, the relations were reduced in 1999.     

Accommodating larger container vessels using an integrated laser system for approach and berthing

Various activities such as dredging and pier lengthening have made it possible for ports to receive larger container vessels, though in many cases this has made berthing the vessels more difficult, necessitating advanced ranging and docking systems. For example, in Koper, Slovenia, navigation through a narrow and bending channel has been made safer through the implementation of a laser-ranging system; a laser docking system is in place so that once the ship is positioning alongside where, say, inadequate fenders await and ship-to-shore (STS) cranes are near, special care is required and parameters such as precise vessel position, transversal speed, and vessel-approaching angle are of great importance. This paper will present the layout of the system and the first results related to the observed docking of large container vessels calling at the Port of Koper.     

A microfluidic platform with integrated flow sensing for focal chemical stimulation of cells and tissue

A microfluidic platform for precise biochemical control of the extracellular microenvironment was developed. A chemical interface was established with cells or tissues through the precise and focal delivery of soluble chemical agents through a pore addressed by a polymer microchannel. Thermal flow sensors were integrated along the length of the microchannel and monitored internal flow rate. Sensor performance was characterized in anticipation of future studies with real-time feedback control of focal delivery. The microfluidic system was characterized by determining the fluid delivery rates through the pores and concentration profiles of agents delivered. Finally, focal delivery to rat retinal tissue was demonstrated.     

An integrated microfluidic system for live bacteria detection from human joint fluid samples by using ethidium monoazide and loop-mediated isothermal amplification

Periprosthetic joint infection (PJI) is one of the severe complications of prosthetic joint replacement. Delayed PJI diagnosis may anchor bacteria in periprosthetic tissues, and removal of the prosthesis might be inevitable. The diagnosis of PJI depends on the identification of microorganisms by standard microbiological cultures or more advanced molecular diagnostic methods for detection of bacterial genes. However, these methods are relatively time-consuming, labor-intensive and not human error-free. Moreover, it is challenging to distinguish live from dead bacteria by using DNA-based molecular diagnostics since bacterial DNA will be remained in the tissue even after the death of the bacteria. In this work, an integrated microfluidic system has been developed to perform the entire molecular diagnostic process for the PJI diagnosis in a single chip. We combined the loop-mediated isothermal amplification (LAMP) with ethidium monoazide (EMA) in an integrated microfluidic system to identify live bacteria with reasonable sensitivity and high specificity. All the diagnostic processes including bacteria isolation, cell lysis, DNA amplification and optical detection can be automatically performed on the integrated microfluidic system by using a compact custom-made control system. The integrated system can accommodate four primers complementary to six regions of the target genes and improve the detection limit by using LAMP. The limit of detection in this multiple EMA-LAMP assay could be as low as 5 fg/reaction (~1 CFU/reaction) when choosing an optimized primer set as we demonstrated in mecA gene detection. Thus, the developed system for PJI diagnosis has great potential to become a point-of-care device.     

Strategic justification of advanced automated technology systems using an expert system

This paper presents an application of expert systems for the strategic justification of advanced automated technology systems. This expert system is developed using the EXSYS expert system shell in the VAX-VMS mainframe environment. It considers important attributes such as suitability (measure of the AMT's compliance with corporate strategy); system capability; performance of the AMT system; and productivity including the financial benefits of the AMT.     

Prediction of river discharge and surface water quality using an integrated geographical information system approach

A methodology is developed for the prediction of river discharge and surface water quality (indexed by nitrogen loading) of a predominantly rural catchment using simple models in an integrated Geographical Information System (GIS). River discharge is predicted using the Soil Conservation Service (SCS) runoff Curve Number model, and surface water quality by the export coefficient model. Main input variable to these models is information on land-use along with ancillary information such as soils. Land-use is an important parameter that affects both discharge and water quality, and it can be derived from classification of remotely sensed images. Unlike conventional models, the models employed here do not require large amounts of data on several hydro-meteorological variables. The models are applied to a rural catchment in eastern England where major land-use changes have occurred in the recent past. Historical land-use data are derived from a variety of sources including maps, aerial photographs and remotely sensed satellite images for various dates ranging from 1931 to 1989. A GIS is a valuable means to enable large amounts of spatial data to be integrated, and to facilitate data manipulation for the specific application of the models. Results are validated using observed runoff and water quality records, and it is shown that the model predictions are of acceptable accuracy. This study demonstrated an application of a GIS to employ simple models to predict river discharge and water quality.     

Development of a yoga posture coaching system using an interactive display based on transfer learning

The Journal of Supercomputing - Yoga is a form of exercise that is beneficial for health, focusing on physical, mental, and spiritual connections. However, practicing yoga and adopting incorrect...     

Model of an integrated intelligent design and manufacturing system

The expert system STATEXS is presented for dimensioning, optimization and manufacture of gears and gearings. The optimum dimensions of the gearing were determined using genetic algorithms, well suited to such problems especially because of their robustness and their ability to detect global extremes. After completion of the calculations and optimization of the gears or gear pairs, there follows one of the most difficult operations, the manufacture of the product with theoretically determined and optimized properties. To this end we have also started to use the genetic algorithm approach for the manufacture of various products with demanding shapes.     

Physical stimulation of mammalian cells using micro-bead impact within a microfluidic environment to enhance growth rate

Various cell stimulation experiments have been traditionally conducted regarding observation of cells to specific stimulative factors, and in light of this area of study, we report a new method by utilizing micro-beads. HeLa cells and MC3T3 cells are cultured in straight PDMS (polydimethylsiloxane) microfluidic devices and are stimulated by sterilized polystyrene micro-beads. Cell culture medium either with or without micro-beads are introduced in microfluidic cell culturing chambers at a specific time interval, and stimulated cells are observed using an inverted microscope. The results show that cells exposed under micro-bead stimulation perform at a higher growth rate than those under normal conditions. This paper demonstrates that micro-beads can be used as a physical stimulation factor and affect cell growth behaviors.     

Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform

This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD_15/45 antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.     

Development and application of an integrated neural system for anHDCL

This study presents the development and industrial application of an integrated neural system in coating weight control for a modern hot dip coating line (HDCL) in a steel mill. The neural system consists of two multilayered feedforward neural networks and a neural adaptive controller. They perform coating weight real-time prediction, feedforward control (FFC), and adaptive feedback control (FBC), respectively. The production line analysis, neural system architecture, learning, associative memories, generalization and real-time applications are addressed in this paper. This integrated neural system has been successfully implemented and applied to an HDCL at Burns Harbor Division, Bethlehem Steel Co., Chesterton, IN. The industrial application results have shown significant improvements in reduction of coating weight transitional footage, variation of the error between the target and actual coating weight, and the coating material used. Some practical aspects for applying a neural system to industrial control are discussed as concluding remarks.