Design and evaluation of a consulting system for database design

Database design is a difficult problem for non-expert designers. It is desirable to assist such designers during the problem solving process by means of a knowledge based (KB) system. A number of prototype KB systems have been proposed, however there are many shortcomings. Few have incorporated sufficient expertise in modeling relationships, particularly higher order relationships. There has been no empirical study that experimentally tested the effectiveness of any of these KB tools. Problem solving behavior of non-experts, whom the systems were intended to assist, has not been one of the bases for system design. In this project a consulting system for conceptual database design that addresses the above short comings was developed and empirically validated.^ The system incorporates (a) findings on why non-experts commit errors and (b) heuristics for modeling relationships. Two approaches to knowledge base implementation--system restrictiveness and decisional guidance--were used and compared in this project. The Restrictive approach is proscriptive and limits the designer's choices at various design phases by forcing him/her to follow a specific design path. The Guidance system approach which is less restrictive, provides context specific, informative and suggestive guidance throughout the design process. The main objectives of the study are to evaluate (1) whether the knowledge-based system is more effective than a system without the knowledge-base and (2) which knowledge implementation--restrictive or guidance--strategy is more effective. To evaluate the effectiveness of the knowledge base itself, the two systems were compared with a system that does not incorporate the expertise (Control).^ The experimental procedure involved the student subjects solving a task without using the system (pre-treatment task) and another task using one of the three systems (experimental task). The experimental task scores of those subjects who performed satisfactorily in the pre-treatment task were analyzed. Results are (1) The knowledge based approach to database design support lead to more accurate solutions than the control system; (2) No significant difference between the two KB approaches; (3) Guidance approach led to best performance; and (4) The subjects perceived the Restrictive system easier to use than the Guidance system. ^

In vivo biocompatibility evaluation of composite polymeric materials for use in a novel artificial heart valve

A novel trileaflet polymer valve is a composite design of a biostable polymer poly(styrene-isobutylene-styrene) (SIBS) with a reinforcement polyethylene terephthalate (PET) fabric. Surface roughness and hydrophilicity vary with fabrication methods and influence leaflet biocompatibility. The purpose of this study was to investigate the biocompatibility of this composite material using both small animal (nonfunctional mode) and large animal (functional mode) models. Composite samples were manufactured using dip coating and solvent casting with different coating thickness (251μm and 50μm). Sample's surface was characterized through qualitative SEM observation and quantitative surface roughness analysis. A novel rat abdominal aorta model was developed to test the composite samples in a similar pulsatile flow condition as its intended use. The sample's tissue response was characterized by histological examination. Among the samples tested, the 25μm solvent-cast sample exhibited the smoothest surface and best biocompatibility in terms of tissue capsulation thickness, and was chosen as the method for fabrication of the SIBS valve. Phosphocholine was used to create a hydrophilic surface on selected composite samples, which resulted in improved blood compatibility. Four SIBS valves (two with phosphocholine modification) were implanted into sheep. Echocardiography, blood chemistry, and system pathology were conducted to evaluate the valve's performance and biocompatibility. No adverse response was identified following implantation. The average survival time was 76 days, and one sheep with the phosphocholine modified valve passed the FDA minimum requirement of 140 days with approximately 20 million cycles of valve activity. The explanted valves were observed under the aid of a dissection microscope, and evaluated via histology, SEM and X-ray. Surface cracks and calcified tissue deposition were found on the leaflets. In conclusion, we demonstrated the applicability of using a new rat abdominal aorta model for biocompatibility assessment of polymeric materials. A smooth and complete coating surface is essential for the biocompatibility of PET/SIBS composite, and surface modification using phosphocholine improves blood compatibility. Extrinsic calcification was identified on the leaflets and was associated with regions of surface cracks.

Design, fabrication, and evaluation of on-chip micro-supercapacitors

Due to the increasing demand for high power and reliable miniaturized energy storage devices, the development of micro-supercapacitors or electrochemical micro-capacitors have attracted much attention in recent years. This dissertation investigates several strategies to develop on-chip micro-supercapacitors with high power and energy density. Micro-supercapacitors based on interdigitated carbon micro-electrode arrays are fabricated through carbon microelectromechanical systems (C-MEMS) technique which is based on carbonization of patterned photoresist. To improve the capacitive behavior, electrochemical activation is performed on carbon micro-electrode arrays. The developed micro-supercapacitors show specific capacitances as high as 75 mFcm-2 at a scan rate of 5 mVs -1 after electrochemical activation for 30 minutes. The capacitance loss is less than 13% after 1000 cyclic voltammetry (CV) cycles. These results indicate that electrochemically activated C-MEMS micro-electrode arrays are promising candidates for on-chip electrochemical micro-capacitor applications. The energy density of micro-supercapacitors was further improved by conformal coating of polypyrrole (PPy) on C-MEMS structures. In these types of micro-devices the three dimensional (3D) carbon microstructures serve as current collectors for high energy density PPy electrodes. The electrochemical characterizations of these micro-supercapacitors show that they can deliver a specific capacitance of about 162.07 mFcm-2 and a specific power of 1.62mWcm -2 at a 20 mVs-1 scan rate. Addressing the need for high power micro-supercapacitors, the application of graphene as electrode materials for micro-supercapacitor was also investigated. The present study suggests a novel method to fabricate graphene-based micro-supercapacitors with thin film or in-plane interdigital electrodes. The fabricated micro-supercapacitors show exceptional frequency response and power handling performance and could effectively charge and discharge at rates as high as 50 Vs-1. CV measurements show that the specific capacitance of the micro-supercapacitor based on reduced graphene oxide and carbon nanotube composites is 6.1 mFcm -2 at scan rate of 0.01Vs-1. At a very high scan rate of 50 Vs-1, a specific capacitance of 2.8 mFcm-2 (stack capacitance of 3.1 Fcm-3) is recorded. This unprecedented performance can potentially broaden the future applications of micro-supercapacitors.

Evaluation of the evidential value of the elemental composition of glass, ink and paper by laser-based microspectrochemical methods

Elemental analysis can become an important piece of evidence to assist the solution of a case. The work presented in this dissertation aims to evaluate the evidential value of the elemental composition of three particular matrices: ink, paper and glass. In the first part of this study, the analytical performance of LIBS and LA-ICP-MS methods was evaluated for paper, writing inks and printing inks. A total of 350 ink specimens were examined including black and blue gel inks, ballpoint inks, inkjets and toners originating from several manufacturing sources and/or batches. The paper collection set consisted of over 200 paper specimens originating from 20 different paper sources produced by 10 different plants. Micro-homogeneity studies show smaller variation of elemental compositions within a single source (i.e., sheet, pen or cartridge) than the observed variation between different sources (i.e., brands, types, batches). Significant and detectable differences in the elemental profile of the inks and paper were observed between samples originating from different sources (discrimination of 87–100% of samples, depending on the sample set under investigation and the method applied). These results support the use of elemental analysis, using LA-ICP-MS and LIBS, for the examination of documents and provide additional discrimination to the currently used techniques in document examination. In the second part of this study, a direct comparison between four analytical methods (µ-XRF, solution-ICP-MS, LA-ICP-MS and LIBS) was conducted for glass analyses using interlaboratory studies. The data provided by 21 participants were used to assess the performance of the analytical methods in associating glass samples from the same source and differentiating different sources, as well as the use of different match criteria (confidence interval (±6s, ±5s, ±4s, ±3s, ±2s), modified confidence interval, t-test (sequential univariate, p=0.05 and p=0.01), t-test with Bonferroni correction (for multivariate comparisons), range overlap, and Hotelling's T2 tests. Error rates (Type 1 and Type 2) are reported for the use of each of these match criteria and depend on the heterogeneity of the glass sources, the repeatability between analytical measurements, and the number of elements that were measured. The study provided recommendations for analytical performance-based parameters for µ-XRF and LA-ICP-MS as well as the best performing match criteria for both analytical techniques, which can be applied now by forensic glass examiners.

Synthesis and Biological Evaluation of Novel Folic Acid Receptor-Targeted, β-Cyclodextrin-Based Drug Complexes for Cancer Treatment

Drug targeting is an active area of research and nano-scaled drug delivery systems hold tremendous potential for the treatment of neoplasms. In this study, a novel cyclodextrin (CD)-based nanoparticle drug delivery system has been assembled and characterized for the therapy of folate receptor-positive [FR(+)] cancer. Water-soluble folic acid (FA)-conjugated CD carriers (FACDs) were successfully synthesized and their structures were confirmed by 1D/2D nuclear magnetic resonance (NMR), matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS), high performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), and circular dichroism. Drug complexes of adamatane (Ada) and cytotoxic doxorubicin (Dox) with FACD were readily obtained by mixed solvent precipitation. The average size of FACD-Ada-Dox was 1.5–2.5 nm. The host-guest association constant Ka was 1,639 M−1 as determined by induced circular dichroism and the hydrophilicity of the FACDs was greatly enhanced compared to unmodified CD. Cellular uptake and FR binding competitive experiments demonstrated an efficient and preferentially targeted delivery of Dox into FR-positive tumor cells and a sustained drug release profile was seen in vitro. The delivery of Dox into FR(+) cancer cells via endocytosis was observed by confocal microscopy and drug uptake of the targeted nanoparticles was 8-fold greater than that of non-targeted drug complexes. Our docking results suggest that FA, FACD and FACD-Ada-Dox could bind human hedgehog interacting protein that contains a FR domain. Mouse cardiomyocytes as well as fibroblast treated with FACD-Ada-Dox had significantly lower levels of reactive oxygen species, with increased content of glutathione and glutathione peroxidase activity, indicating a reduced potential for Dox-induced cardiotoxicity. These results indicate that the targeted drug complex possesses high drug association and sustained drug release properties with good biocompatibility and physiological stability. The novel FA-conjugated β-CD based drug complex might be promising as an anti-tumor treatment for FR(+) cancer.

The effect of dual -language instruction on academic achievement as measured by the SAT: An evaluation of the "International Studies Program" of Miami -Dade County Public Schools

This study investigated the influence that receiving instruction in two languages, English and Spanish, had on the performance of students enrolled in the International Studies Program (delayed partial immersion model) of Miami Dade County Public Schools on a standardized test in English, the Stanford Achievement Test, eighth edition, for three of its sections, Reading Comprehension, Mathematics Computations, and Mathematics Applications.^ The performance of the selected IS program/Spanish section cohort of students (N = 55) on the SAT Reading Comprehension, Mathematics Computation, and Mathematics Application along four consecutive years was contrasted with that of a control group of comparable students selected within the same feeder pattern where the IS program is implemented (N = 21). The performance of the group was also compared to the cross-sectional achievement patterns of the school's corresponding feeder pattern, region, and district.^ The research model for the study was a variation of the "causal-comparative" or "ex post facto design" sometimes referred to as "prospective". After data were collected from MDCPS, t-tests were performed to compare IS-Spanish students SAT performance for grades 3 to 6 for years 1994 to 1997 to control group, feeder pattern, region and district norms for each year for Reading Comprehension, Mathematics Computation, and Mathematics Applications. Repeated measures ANOVA and Tukey's tests were calculated to compare the mean percentiles of the groups under study and the possible interactions of the different variables. All tests were performed at the 5% significance level.^ From the analyses of the tests it was deduced that the IS group performed significantly better than the control group for all the three measures along the four years. The IS group mean percentiles on the three measures were also significantly higher than those of the feeder pattern, region, and district. The null hypotheses were rejected and it was concluded that receiving instruction in two languages did not negatively affect the performance of IS program students on tests taken in English. It was also concluded that the particular design the IS program enhances the general performance of participant students on Standardized tests.^ The quantitative analyses were coupled with interviews from teachers and administrators of the IS program to gain additional insight about different aspects of the implementation of the program at each particular school. ^

Evaluation of non-contact sampling and detection of explosives using receiver operating characteristic curves

The growing need for fast sampling of explosives in high throughput areas has increased the demand for improved technology for the trace detection of illicit compounds. Detection of the volatiles associated with the presence of the illicit compounds offer a different approach for sensitive trace detection of these compounds without increasing the false positive alarm rate. This study evaluated the performance of non-contact sampling and detection systems using statistical analysis through the construction of Receiver Operating Characteristic (ROC) curves in real-world scenarios for the detection of volatiles in the headspace of smokeless powder, used as the model system for generalizing explosives detection. A novel sorbent coated disk coined planar solid phase microextraction (PSPME) was previously used for rapid, non-contact sampling of the headspace containers. The limits of detection for the PSPME coupled to IMS detection was determined to be 0.5-24 ng for vapor sampling of volatile chemical compounds associated with illicit compounds and demonstrated an extraction efficiency of three times greater than other commercially available substrates, retaining >50% of the analyte after 30 minutes sampling of an analyte spike in comparison to a non-detect for the unmodified filters. Both static and dynamic PSPME sampling was used coupled with two ion mobility spectrometer (IMS) detection systems in which 10-500 mg quantities of smokeless powders were detected within 5-10 minutes of static sampling and 1 minute of dynamic sampling time in 1-45 L closed systems, resulting in faster sampling and analysis times in comparison to conventional solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis. Similar real-world scenarios were sampled in low and high clutter environments with zero false positive rates. Excellent PSPME-IMS detection of the volatile analytes were visualized from the ROC curves, resulting with areas under the curves (AUC) of 0.85-1.0 and 0.81-1.0 for portable and bench-top IMS systems, respectively. Construction of ROC curves were also developed for SPME-GC-MS resulting with AUC of 0.95-1.0, comparable with PSPME-IMS detection. The PSPME-IMS technique provides less false positive results for non-contact vapor sampling, cutting the cost and providing an effective sampling and detection needed in high-throughput scenarios, resulting in similar performance in comparison to well-established techniques with the added advantage of fast detection in the field.

TEACHNJ: An Evaluation of Two Years Implementation

The Teacher Effectiveness and Accountability for the Children of New Jersey (TEACHNJ) Act was adopted by the New Jersey legislature in August 2012 with the intent to raise student achievement by improving the overall quality of instruction. As a result of this act, new teacher evaluation systems are being introduced in school districts across the state in an effort to more accurately assess teacher performance. The new teacher evaluations will be based on multiple classroom observations as well as the academic achievement of their students as measured on standardized tests. In addition, professional development opportunities are likely to change under this legislation, with schools customizing professional development programs to more effectively meet the needs of their teachers. The overarching question that informs our research is what impact will TEACH NJ have on the overall value of teacher evaluations and the quality of professional development opportunities offered to teachers. Data collected through survey research presents the pre-implementation practices (2011-2012 school year) as well as one year post-implementation practices (2013-2014) taking place in school districts throughout New Jersey. The findings reflect teachers’ perceptions of the value of the current teacher evaluation practices, the quality of the current professional development opportunities and the value the school administration places on teacher evaluations.

An evaluation of a curriculum response to the State of Florida mandate for computer literacy at a large comprehensive high school in Dade County, Florida

Minimum Student Performance Standards in Computer Literacy and Science were passed by the Florida Legislature through the Educational Reform Act of 1983. This act mandated that all Florida high school graduates receive training in computer literacy. Schools and school systems were charged with the task of determining the best methods to deliver this instruction to their students. The scope of this study is to evaluate one school's response to the state of Florida's computer literacy mandate. The study was conducted at Miami Palmetto Senior High School, located in Dade County, Florida. The administration of Miami Palmetto Senior High School chose to develop and implement a new program to comply with the state mandate - integrating computer literacy into the existing biology curriculum. The study evaluated the curriculum to determine if computer literacy could be integrated successfully and meet both the biology and computer literacy objectives. The findings in this study showed that there were no significant differences between biology scores of the students taking the integrated curriculum and those taking a traditional curriculum of biology. Student in the integrated curriculum not only met the biology objectives as well as those in the traditional curriculum, they also successfully completed the intended objectives for computer literacy. Two sets of objectives were successfully completed in the integrated classes in the same amount of time used to complete one set of objectives in the traditional biology classes. Therefore, integrated curriculum was the more efficient means of meeting the intended objectives of both biology and computer literacy.

Design and evaluation of a consulting system for database design

Database design is a difficult problem for non-expert designers. It is desirable to assist such designers during the problem solving process by means of a knowledge based (KB) system. Although a number of prototype KB systems have been proposed, there are many shortcomings. Firstly, few have incorporated sufficient expertise in modeling relationships, particularly higher order relationships. Secondly, there does not seem to be any published empirical study that experimentally tested the effectiveness of any of these KB tools. Thirdly, problem solving behavior of non-experts, whom the systems were intended to assist, has not been one of the bases for system design. In this project, a consulting system, called CODA, for conceptual database design that addresses the above short comings was developed and empirically validated. More specifically, the CODA system incorporates (a) findings on why non-experts commit errors and (b) heuristics for modeling relationships. Two approaches to knowledge base implementation were used and compared in this project, namely system restrictiveness and decisional guidance (Silver 1990). The Restrictive system uses a proscriptive approach and limits the designer's choices at various design phases by forcing him/her to follow a specific design path. The Guidance system approach, which is less restrictive, involves providing context specific, informative and suggestive guidance throughout the design process. Both the approaches would prevent erroneous design decisions. The main objectives of the study are to evaluate (1) whether the knowledge-based system is more effective than the system without a knowledge-base and (2) which approach to knowledge implementation - whether Restrictive or Guidance - is more effective. To evaluate the effectiveness of the knowledge base itself, the systems were compared with a system that does not incorporate the expertise (Control). An experimental procedure using student subjects was used to test the effectiveness of the systems. The subjects solved a task without using the system (pre-treatment task) and another task using one of the three systems, viz. Control, Guidance or Restrictive (experimental task). Analysis of experimental task scores of those subjects who performed satisfactorily in the pre-treatment task revealed that the knowledge based approach to database design support lead to more accurate solutions than the control system. Among the two KB approaches, Guidance approach was found to lead to better performance when compared to the Control system. It was found that the subjects perceived the Restrictive system easier to use than the Guidance system.

An evaluation of home range models for marine fish tracking and fine scale habitat use and movement patterns of age 1 Greenland cod (Gadus macrocephalus ogac)

My thesis examines fine-scale habitat use and movement patterns of age 1 Greenland cod (Gadus macrocephalus ogac) tracked using acoustic telemetry. Recent advances in tracking technologies such as GPS and acoustic telemetry have led to increasingly large and detailed datasets that present new opportunities for researchers to address fine-scale ecological questions regarding animal movement and spatial distribution. There is a growing demand for home range models that will not only work with massive quantities of autocorrelated data, but that can also exploit the added detail inherent in these high-resolution datasets. Most published home range studies use radio-telemetry or satellite data from terrestrial mammals or avian species, and most studies that evaluate the relative performance of home range models use simulated data. In Chapter 2, I used actual field-collected data from age-1 Greenland cod tracked with acoustic telemetry to evaluate the accuracy and precision of six home range models: minimum convex polygons, kernel densities with plug-in bandwidth selection and the reference bandwidth, adaptive local convex hulls, Brownian bridges, and dynamic Brownian bridges. I then applied the most appropriate model to two years (2010-2012) of tracking data collected from 82 tagged Greenland cod tracked in Newman Sound, Newfoundland, Canada, to determine diel and seasonal differences in habitat use and movement patterns (Chapter 3). Little is known of juvenile cod ecology, so resolving these relationships will provide valuable insight into activity patterns, habitat use, and predator-prey dynamics, while filling a knowledge gap regarding the use of space by age 1 Greenland cod in a coastal nursery habitat. By doing so, my thesis demonstrates an appropriate technique for modelling the spatial use of fish from acoustic telemetry data that can be applied to high-resolution, high-frequency tracking datasets collected from mobile organisms in any environment.

Investigation on performance characteristics of dense graded hot mix asphalt with RAP and crumb rubber

The objective of this study was to fundamentally characterize the laboratory performance of traditional hot mix asphalt (HMA) mixtures incorporating high RAP content and waste tire crumb rubber through their fundamental engineering properties. The nominal maximum aggregates size was chosen for this research was 12mm (considering the limitation of aggregate size for surface layer) and both coarse and fine aggregates are commonly used in Italy that were examined and analyzed in this study. On the other hand, the RAP plays an important role in reducing production costs and enhancing the environmentally sustainable pavements instead of using virgin materials in HMA. Particularly, this study has aimed to use 30% of RAP content (25% fine aggregate RAP and 5% coarse aggregate RAP) and 1% of CR additives by the total weight of aggregates for mix design. The mixture of aggregates, RAP and CR were blended with different amount of unmodified binder through dry processes. Generally, the main purposes of this study were investigating on capability of using RAP and CR in dense graded HMA and comparing the performance of rejuvenator in RAP with CR. In addition, based on the engineering analyses during the study, we were able compare the fundamental Indirect Tensile Strength (ITS) value of dense graded HMA and also mechanical characteristics in terms of Indirect Tensile Stiffness Modulus (ITSM). In order to get an extended comparable data, four groups of different mixtures such as conventional mixture with only virgin aggregates (DV), mixture with RAP (DR), mixture with RAP and rejuvenator (DRR), and mixture with RAP, rejuvenator, CR (DRRCr) were investigated in this research experimentally. Finally, the results of those tests indicated that the mixtures with RAP and CR had the high stiffness and less thermal sensitivity, while the mixture with virgin aggregates only had very low values in comparison.

Evaluation of semi-passive treatment technologies for septic lagoon capacity expansion

In Canada, increases in rural development has led to a growing need to effectively manage the resulting municipal and city sewage without the addition of significant cost- and energy- expending infrastructure. Storring Septic Service Limited is a family-owned, licensed wastewater treatment facility located in eastern Ontario. It makes use of a passive waste stabilization pond system to treat and dispose of waste and wastewater in an environmentally responsible manner. Storring Septic, like many other similar small-scale wastewater treatment facilities across Canada, has the potential to act as a sustainable eco-engineered facility that municipalities and service providers could utilize to manage and dispose of their wastewater. However, it is of concern that the substantial inclusion of third party material could be detrimental to the stability and robustness of the pond system. In order to augment the capacity of the current facility, and ensure it remains a self-sustaining system with the capacity to safely accept septage from other sewage haulers, it was hypothesized that pond effluent treatment could be further enhanced through the incorporation of one of three different technology solutions, which would allow the reduction of wastewater quality parameters below existing regulatory effluent discharge limits put in place by Ontario’s Ministry of the Environment and Climate Change (MOECC). Two of these solutions make use of biofilm technologies in order to enhance the removal of wastewater parameters of interest, and the third utilizes the natural water filtration capabilities of zebra mussels. Pilot-scale testing investigated the effects of each of these technologies on treatment performance under both cold and warm weather operation. This research aimed to understand the important mechanisms behind biological filtration methods in order to choose and optimize the best treatment strategy for full-scale testing and implementation. In doing so, a recommendation matrix was elaborated provided with the potential to be used as a universal operational strategy for wastewater treatment facilities located in environments of similar climate and ecology.

Design, analysis and evaluation of sigma-delta based beamformers for medical ultrasound imaging applications

The inherent analogue nature of medical ultrasound signals in conjunction with the abundant merits provided by digital image acquisition, together with the increasing use of relatively simple front-end circuitries, have created considerable demand for single-bit  beamformers in digital ultrasound imaging systems. Furthermore, the increasing need to design lightweight ultrasound systems with low power consumption and low noise, provide ample justification for development and innovation in the use of single-bit  beamformers in ultrasound imaging systems. The overall aim of this research program is to investigate, establish, develop and confirm through a combination of theoretical analysis and detailed simulations, that utilize raw phantom data sets, suitable techniques for the design of simple-to-implement hardware efficient  digital ultrasound beamformers to address the requirements for 3D scanners with large channel counts, as well as portable and lightweight ultrasound scanners for point-of-care applications and intravascular imaging systems. In addition, the stability boundaries of higher-order High-Pass (HP) and Band-Pass (BP) Σ−Δ modulators for single- and dual- sinusoidal inputs are determined using quasi-linear modeling together with the describing-function method, to more accurately model the  modulator quantizer. The theoretical results are shown to be in good agreement with the simulation results for a variety of input amplitudes, bandwidths, and modulator orders. The proposed mathematical models of the quantizer will immensely help speed up the design of higher order HP and BP Σ−Δ modulators to be applicable for digital ultrasound beamformers. Finally, a user friendly design and performance evaluation tool for LP, BP and HP  modulators is developed. This toolbox, which uses various design methodologies and covers an assortment of  modulators topologies, is intended to accelerate the design process and evaluation of  modulators. This design tool is further developed to enable the design, analysis and evaluation of  beamformer structures including the noise analyses of the final B-scan images. Thus, this tool will allow researchers and practitioners to design and verify different reconstruction filters and analyze the results directly on the B-scan ultrasound images thereby saving considerable time and effort.

Inter-Laboratory Evaluation of a Next-Generation Sequencing Panel for Acute Myeloid Leukemia

INTRODUCTION: Acute myeloid leukemia (AML) is a heterogeneous clonal disorder often associated with dismal overall survival. The clinical diversity of AML is reflected in the range of recurrent somatic mutations in several genes, many of which have a prognostic and therapeutic value. Targeted next-generation sequencing (NGS) of these genes has the potential for translation into clinical practice. In order to assess this potential, an inter-laboratory evaluation of a commercially available AML gene panel across three diagnostic centres in the UK and Ireland was performed.

METHODS: DNA from six AML patient samples was distributed to each centre and processed using a standardised workflow, including a common sequencing platform, sequencing chips and bioinformatics pipeline. A duplicate sample in each centre was run to assess inter- and intra-laboratory performance.

RESULTS: An average sample read depth of 2725X (range 629-5600) was achieved using six samples per chip, with some variability observed in the depth of coverage generated for individual samples and between centres. A total of 16 somatic mutations were detected in the six AML samples, with a mean of 2.7 mutations per sample (range 1-4) representing nine genes on the panel. 15/16 mutations were identified by all three centres. Allelic frequencies of the mutations ranged from 5.6 to 53.3 % (median 44.4 %), with a high level of concordance of these frequencies between centres, for mutations detected.

CONCLUSION: In this inter-laboratory comparison, a high concordance, reproducibility and robustness was demonstrated using a commercially available NGS AML gene panel and platform.