Original Pool Research Articles

Setting health research priorities using the CHNRI method: VI. Quantitative properties of human collective opinion.

IntroductionCrowdsourcing has become an increasingly important tool to address many problems – from government elections in democracies, stock market prices, to modern online tools such as TripAdvisor or Internet Movie Database (IMDB). The CHNRI method (the acronym for the Child Health and Nutrition Research Initiative) for setting health research priorities has crowdsourcing as the major component, which it uses to generate, assess and prioritize between many competing health research ideas.MethodsWe conducted a series of analyses using data from a group of 91 scorers to explore the quantitative properties of their collective opinion. We were interested in the stability of their collective opinion as the sample size increases from 15 to 90. From a pool of 91 scorers who took part in a previous CHNRI exercise, we used sampling with replacement to generate multiple random samples of different size. First, for each sample generated, we identified the top 20 ranked research ideas, among 205 that were proposed and scored, and calculated the concordance with the ranking generated by the 91 original scorers. Second, we used rank correlation coefficients to compare the ranks assigned to all 205 proposed research ideas when samples of different size are used. We also analysed the original pool of 91 scorers to to look for evidence of scoring variations based on scorers' characteristics.ResultsThe sample sizes investigated ranged from 15 to 90. The concordance for the top 20 scored research ideas increased with sample sizes up to about 55 experts. At this point, the median level of concordance stabilized at 15/20 top ranked questions (75%), with the interquartile range also generally stable (14–16). There was little further increase in overlap when the sample size increased from 55 to 90. When analysing the ranking of all 205 ideas, the rank correlation coefficient increased as the sample size increased, with a median correlation of 0.95 reached at the sample size of 45 experts (median of the rank correlation coefficient = 0.95; IQR 0.94–0.96).ConclusionsOur analyses suggest that the collective opinion of an expert group on a large number of research ideas, expressed through categorical variables (Yes/No/Not Sure/Don't know), stabilises relatively quickly in terms of identifying the ideas that have most support. In the exercise we found a high degree of reproducibility of the identified research priorities was achieved with as few as 45–55 experts.

Targeted deep resequencing of ALOX5 and ALOX5AP in patients with diabetes and association of rare variants with leukotriene pathways.

The aim of the present study was to investigate a possible association between the accumulation of rare coding variants in the genes for arachidonate 5-lipoxygenase (ALOX5) and ALOX5-activating protein (ALOX5AP), and corresponding production of leukotrienes (LTs) in patients with type 2 diabetes mellitus (T2DM) receiving acetylsalicylic therapy. Twenty exons and corresponding introns of the selected genes were resequenced in 303 DNA samples from patients with T2DM using pooled polymerase chain reaction amplification and next-generation sequencing, using an Illumina HiSeq 2000 sequencing system. The observed non-synonymous variants were further confirmed by individual genotyping of DNA samples comprising of all individuals from the original discovery pools. The association between the investigated phenotypes was based on LTB4 and LTE4 concentrations, and the accumulation of rare missense variants (genetic burden) in investigated genes was evaluated using statistical collapsing tests. A total of 10 exonic variants were identified for each resequenced gene, including 5 missense and 5 synonymous variants. The rare missense variants did not exhibit statistically significant differences in the accumulation pattern between the patients with low and high LTs concentrations. As the present study only included patients with T2DM, it is unclear whether the absence of observed association between the accumulation of rare missense variants in investigated genes and LT production is associated with diabetic populations only or may also be applied to other populations.

Orally Ingested Transient Receptor Potential (TRP) Channel Activators Attenuate the Intensity‐Duration of Voluntarily Induced Muscle Cramps in Humans

Transient receptor potential (TRP) ion channels in the mouth, esophagus and stomach are activated by ingestion of many naturally occurring food extracts. Recent evidence suggests that oral ingestion of TRPA1 and TRPV1 channel agonists may reflexly attenuate the intensity and/or duration of electrically induced muscle cramps in humans (Short et al., 2014, 2015; Hegarty et al., 2015), presumably by dampening alpha motor neuron excitability. To test this effect on voluntarily initiated calf muscle cramps, we conducted a randomized, double‐blinded, vehicle‐controlled crossover study. Subjects performed 3 baseline experiments in which they underwent a voluntary exercise‐induced muscle cramp protocol. From an original pool of 34 volunteers with a self‐reported history of exercise‐related muscle cramping, 6 men and 14 women (age: 22±3 yrs representing 59% of screened subjects) were able to consistently reproduce cramps voluntarily. Each of the 20 enrolled subjects subsequently participated in 2 experimental trials, one 15 min after consuming less than 3 oz. of an active (A) formulation containing a mixture of TRPA1 and TRPV1 activators derived from natural extracts and another 15 min after ingesting a vehicle control (V) in a randomized sequence. EMG leads were placed on the medial gastrocnemius, lateral gastrocnemius, soleus, and tibia. The supine subject's foot was securely strapped to an adjustable force transducer and the knee flexed to position the foot at the angle of maximum plantar flexion. As directed by the experimenter, the subject then attempted to produce a cramp by first pre‐shortening the gastrocnemius, isolating the triceps surae, and then performing a maximal voluntary static contraction (MVC). Contractions were held for 90 s or until a cramp occurred. If no cramp occurred, subjects rested for 10 min before attempting to elicit a cramp for a second time. Subjects repeated this pattern for up to 5 attempts within any given experiment; all experiments were separated by at least 1 week. EMG and force were recorded at baseline, during the static muscle contraction, and throughout the duration of the muscle cramp. There were no significant differences between active and vehicle control for pre‐cramp MVC (A: 12.4±7.0 (SD) kg, V: 12.5±7.6 kg; p=0.99), post‐cramp MVC (A: 10.1±5.6 kg, V: 10.7±6.8 kg; p=0.40), pre‐cramp contraction duration (A: 32.7±21.8 s, V: 28.3±17.2 s; p=0.16), or cramp duration (A: 13.7±11.4 s, V: 16.9±16.3 s; p=0.08). However, the integrated EMG signal, i.e., the area under the EMG intensity‐duration curve, was significantly lower (p=0.01) for A (0.51±0.38 V•s) than V (0.80±0.69 V•s). Composite subjective ratings of soreness (on a 1 to 10 rating scale) during the initial 20 min after cessation of cramping were also significantly (p=0.02) lower for the active formulation (3.4±4.6) than the vehicle control (4.0±2.2). Consistent with results from studies that have evoked leg muscle cramps using electrical stimulation, oral TRP channel activators significantly attenuated the intensity‐duration profile of voluntarily initiated muscle cramps in this sample of young adults.Support or Funding InformationFUNDING: Flex Pharma, Inc.

Targeting Insulin Receptor with a Novel Internalizing Aptamer

Nucleic acid-based aptamers are emerging as therapeutic antagonists of disease-associated proteins such as receptor tyrosine kinases. They are selected by an in vitro combinatorial chemistry approach, named Systematic Evolution of Ligands by Exponential enrichment (SELEX), and thanks to their small size and unique chemical characteristics, they possess several advantages over antibodies as diagnostics and therapeutics. In addition, aptamers that rapidly internalize into target cells hold as well great potential for their in vivo use as delivery tools of secondary therapeutic agents. Here, we describe a nuclease resistant RNA aptamer, named GL56, which specifically recognizes the insulin receptor (IR). Isolated by a cell-based SELEX method that allows enrichment for internalizing aptamers, GL56 rapidly internalizes into target cells and is able to discriminate IR from the highly homologous insulin-like growth factor receptor 1. Notably, when applied to IR expressing cancer cells, the aptamer inhibits IR dependent signaling. Given the growing interest in the insulin receptor as target for cancer treatment, GL56 reveals a novel molecule with great translational potential as inhibitor and delivery tool for IR-dependent cancers.

편평족 중학생의 고관절 교정 운동 프로그램과 발교정구 착용 유무가 안정시 종골 기립 각도, 발목의 가동범위, 코어 근력에 미치는 영향

Objective : The purpose of this study is to evaluate the effects of hip joint exercises and orthotics on RCSP, ankle's range of motion, and core muscle strength of middle school students with pes planus. Method : Out of the original pool of 200 students, 60 students with pes planus (RCSP < -2) were selected for the study. The selected 60 students were then divided into four groups. The first group was a combined orthotics and exercise group (12 students), the second was the orthotics-only group (9 students), the third was the exercise-only group (8 students), and the last was the control group (10 students). Exercise groups worked out twice a week for 60 minutes per session over 8 weeks. The independent variables were corrective hip joint exercises and orthotics. The dependant variables consisted of kinematic and kinetic variables. The kinematic variables were RCSP, and ankle's range of motion (dorsiflexion and plantarflexion). The kinetic variables were muscles forces that consist in core muscle strength, which are hip joint adduction, abduction, and flexion muscles forces. Statistical analysis was performed via SPSS 18.0 with multivariate analysis of covariance (MANCOVA) and a paired t-test was used. Results : The left foot was more responsive to the treatments, both exercise and orthotics, than the right foot. RCSP improved significantly in the left foot for the first and third groups. Only the first group significantly improved hip joint adduction, abduction, and flexion muscles' strengths. As for the ankle's range of motion of the left foot, plantarflexion showed improvement when treated with exercise, orthotics, or both. Conclusion : This study found that exercise is more effective in correcting RCSP and foot orthotics is more effective in reinforcing core muscle strength. Future studies should expand on these results to examine the relationship between the ankle, hip, and pelvis.

Face identification with second-order pooling in single-layer networks

Automatic face recognition has received significant performance improvement by developing specialized facial image representations. On the other hand, spatial pyramid pooling of features encoded by an over-complete dictionary has been the key component of many state-of-the-art generic objective classification systems. Inspired by its success, in this work we develop a new face image representation method under the framework of single-layer networks, where the key component is the second-order pooling layer. The proposed method differs from the previous methods in that, we encode the densely extracted local patches by a small-size dictionary; and the facial image signatures are obtained by pooling the second-order statistics of the encoded features. We show the importance of the encoding procedure, which is bypassed by the original second-order pooling method to avoid the high computational cost. Equipped with a simple linear classifier, the proposed method outperforms the state-of-the-art face identification performance by large margins. For example, on the LFW databases, the proposed method performs better than the previous best by around 13% accuracy.

Next-generation re-sequencing of genes involved in increased platelet reactivity in diabetic patients on acetylsalicylic acid

The objective of this study was to investigate whether rare missense genetic variants in several genes related to platelet functions and acetylsalicylic acid (ASA) response are associated with the platelet reactivity in patients with diabetes type 2 (T2D) on ASA therapy. Fifty eight exons and corresponding introns of eight selected genes, including PTGS1, PTGS2, TXBAS1, PTGIS, ADRA2A, ADRA2B, TXBA2R, and P2RY1 were re-sequenced in 230 DNA samples from T2D patients by using a pooled PCR amplification and next-generation sequencing by Illumina HiSeq2000. The observed non-synonymous variants were confirmed by individual genotyping of 384 DNA samples comprising of the individuals from the original discovery pools and additional verification cohort of 154 ASA-treated T2DM patients. The association between investigated phenotypes (ASA induced changes in platelets reactivity by PFA-100, VerifyNow and serum thromboxane B2 level [sTxB2]), and accumulation of rare missense variants (genetic burden) in investigated genes was tested using statistical collapsing tests. We identified a total of 35 exonic variants, including 3 common missense variants, 15 rare missense variants, and 17 synonymous variants in 8 investigated genes. The rare missense variants exhibited statistically significant difference in the accumulation pattern between a group of patients with increased and normal platelet reactivity based on PFA-100 assay. Our study suggests that genetic burden of the rare functional variants in eight genes may contribute to differences in the platelet reactivity measured with the PFA-100 assay in the T2DM patients treated with ASA.

Genome-wide mapping reveals single-origin chromosome replication in Leishmania, a eukaryotic microbe.

BackgroundDNA replication initiates on defined genome sites, termed origins. Origin usage appears to follow common rules in the eukaryotic organisms examined to date: all chromosomes are replicated from multiple origins, which display variations in firing efficiency and are selected from a larger pool of potential origins. To ask if these features of DNA replication are true of all eukaryotes, we describe genome-wide origin mapping in the parasite Leishmania.ResultsOrigin mapping in Leishmania suggests a striking divergence in origin usage relative to characterized eukaryotes, since each chromosome appears to be replicated from a single origin. By comparing two species of Leishmania, we find evidence that such origin singularity is maintained in the face of chromosome fusion or fission events during evolution. Mapping Leishmania origins suggests that all origins fire with equal efficiency, and that the genomic sites occupied by origins differ from related non-origins sites. Finally, we provide evidence that origin location in Leishmania displays striking conservation with Trypanosoma brucei, despite the latter parasite replicating its chromosomes from multiple, variable strength origins.ConclusionsThe demonstration of chromosome replication for a single origin in Leishmania, a microbial eukaryote, has implications for the evolution of origin multiplicity and associated controls, and may explain the pervasive aneuploidy that characterizes Leishmania chromosome architecture.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0788-9) contains supplementary material, which is available to authorized users.

Landscape genomics reveal signatures of local adaptation in barley (Hordeum vulgare L.).

Land plants are sessile organisms that cannot escape the adverse climatic conditions of a given environment. Hence, adaptation is one of the solutions to surviving in a challenging environment. This study was aimed at detecting adaptive loci in barley landraces that are affected by selection. To that end, a diverse population of barley landraces was analyzed using the genotyping by sequencing approach. Climatic data for altitude, rainfall and temperature were collected from 61 weather sites near the origin of selected landraces across Ethiopia. Population structure analysis revealed three groups whereas spatial analysis accounted significant similarities at shorter geographic distances (< 40 Km) among barley landraces. Partitioning the variance between climate variables and geographic distances indicated that climate variables accounted for most of the explainable genetic variation. Markers by climatic variables association analysis resulted in altogether 18 and 62 putative adaptive loci using Bayenv and latent factor mixed model (LFMM), respectively. Subsequent analysis of the associated SNPs revealed putative candidate genes for plant adaptation. This study highlights the presence of putative adaptive loci among barley landraces representing original gene pool of the farming communities.

Parsimonious Description of Glucose Variability in Type 2 Diabetes by Sparse Principal Component Analysis.

Abnormal glucose variability (GV) is a risk factor for diabetes complications, and tens of indices for its quantification from continuous glucose monitoring (CGM) time series have been proposed. However, the information carried by these indices is redundant, and a parsimonious description of GV can be obtained through sparse principal component analysis (SPCA). We have recently shown that a set of 10 metrics selected by SPCA is able to describe more than 60% of the variance of 25 GV indicators in type 1 diabetes (T1D). Here, we want to extend the application of SPCA to type 2 diabetes (T2D). A data set of CGM time series collected in 13 T2D subjects was considered. The 25 GV indices considered for T1D were evaluated. SPCA was used to select a subset of indices able to describe the majority of the original variance. A subset of 10 indicators was selected and allowed to describe 83% of the variance of the original pool of 25 indices. Four metrics sufficient to describe 67% of the original variance turned out to be shared by the parsimonious sets of indices in T1D and T2D. Starting from a pool of 25 indices assessed from CGM time series in T2D subjects, reduced subsets of metrics virtually providing the same information content can be determined by SPCA. The fact that these indices also appear in the parsimonious description of GV in T1D may indicate that they could be particularly informative of GV in diabetes, regardless of the specific type of disease.

Creating the Nurses' Environmental Awareness Tool (NEAT).

Acute care delivery creates secondary health risks to patients, health care workers, and the environment through a complex waste stream, intensive energy use, and frequent use of harmful chemicals. Nurses are among the most affected by these risks and are also pivotal change agents in reducing the negative impacts of health care delivery. Assessing nurses' understanding of health care-associated environmental health risks is essential if care is to be delivered in an environmentally safe and healthy manner, as indicated by published professional standards of nursing practice. However, psychometrically sound instruments that measure nurses' awareness of the environmental impacts of nursing practice are not available. To address this gap, a prototype of the Nurse's Environmental Awareness Tool (NEAT) was developed. Seven content experts in environmental health nursing and/or psychometrics were asked to review draft items. Comments were analyzed and applied to the scale items. Several revisions from the original item pool were made. The resulting draft NEAT includes six subscales, in three paired subsets. This article provides a summary of the process of item development and scale design. These findings, although preliminary, provide a foundation for subsequent psychometric testing. The result of this study is the creation of an instrument to measure nurses' awareness of and behaviors associated with the environmental impact of their practices.

Life‐history traits and geographical divergence in wild rice ( Oryza rufipogon ) gene pool in Indochina Peninsula region

Indochina Peninsula is the primary centre of diversity of rice and lies partly in the centre of origin of cultivated rice (Oryza sativa) where the wild ancestor (Oryza rufipogon) is still abundant. The wild gene pool is potentially endangered by urbanisation and the expansion of agriculture, and by introgression hybridisation with locally cultivated rice varieties. To determine genetic diversity and structure of the wild rice of the region we genotyped nearly 1000 individuals using 20 microsatellite loci. We found ecological differentiation in 48 populations, distinguishable by their life-history traits and the country of origin. Geographical divergence was suggested by isolation of the perennial Myanmar populations from those of Cambodia, Laos and Thailand. The annual types would be most likely to have lost genetic variation because of genetic drift and inbreeding. The growing of cultivated and wild rice together, however, gives ample opportunities for hybridisation, which already shows signs of genetic mixing, and will ultimately lead to replacement of the original wild rice gene pool. For conservation we suggest that wild rice should be conserved ex situ in order to prevent introgression from cultivated rice, along with in situ conservation in individual countries for the recurrent evolutionary process through local adaptation, but with sufficient isolation from cultivated rice fields to preserve genetic integrity of the wild populations.

PIWI Slicing and RNA Elements in Precursors Instruct Directional Primary piRNA Biogenesis.

PIWI proteins and PIWI-interacting RNAs (piRNAs) mediate repression of transposons in the animal gonads. Primary processing converts long single-stranded RNAs into ∼30-nt piRNAs, but their entry into the biogenesis pathway is unknown. Here, we demonstrate that an RNA element at the 5' end of a piRNA cluster—which we termed piRNA trigger sequence (PTS)—can induce primary processing of any downstream sequence. We propose that such signals are triggers for the generation of the original pool of piRNAs. We also demonstrate that endonucleolytic cleavage of a transcript by a cytosolic PIWI results in its entry into primary processing, which triggers the generation of non-overlapping, contiguous primary piRNAs in the 3' direction from the target transcript. These piRNAs are loaded into a nuclear PIWI, thereby linking cytoplasmic post-transcriptional silencing to nuclear transcriptional repression.

Deconstructing the polymerase chain reaction: understanding and correcting bias associated with primer degeneracies and primer-template mismatches.

The polymerase chain reaction (PCR) is sensitive to mismatches between primer and template, and mismatches can lead to inefficient amplification of targeted regions of DNA template. In PCRs in which a degenerate primer pool is employed, each primer can behave differently. Therefore, inefficiencies due to different primer melting temperatures within a degenerate primer pool, in addition to mismatches between primer binding sites and primers, can lead to a distortion of the true relative abundance of targets in the original DNA pool. A theoretical analysis indicated that a combination of primer-template and primer-amplicon interactions during PCR cycles 3–12 is potentially responsible for this distortion. To test this hypothesis, we developed a novel amplification strategy, entitled “Polymerase-exonuclease (PEX) PCR”, in which primer-template interactions and primer-amplicon interactions are separated. The PEX PCR method substantially and significantly improved the evenness of recovery of sequences from a mock community of known composition, and allowed for amplification of templates with introduced mismatches near the 3’ end of the primer annealing sites. When the PEX PCR method was applied to genomic DNA extracted from complex environmental samples, a significant shift in the observed microbial community was detected. Furthermore, the PEX PCR method provides a mechanism to identify which primers in a primer pool are annealing to target gDNA. Primer utilization patterns revealed that at high annealing temperatures in the PEX PCR method, perfect match annealing predominates, while at lower annealing temperatures, primers with up to four mismatches with templates can contribute substantially to amplification. The PEX PCR method is simple to perform, is limited to PCR mixes and a single exonuclease step which can be performed without reaction cleanup, and is recommended for reactions in which degenerate primer pools are used or when mismatches between primers and template are possible.

The DNA repair endonuclease Mus81 facilitates fast DNA replication in the absence of exogenous damage.

The Mus81 endonuclease resolves recombination intermediates and mediates cellular responses to exogenous replicative stress. Here, we show that Mus81 also regulates the rate of DNA replication during normal growth by promoting replication fork progression while reducing the frequency of replication initiation events. In the absence of Mus81 endonuclease activity, DNA synthesis is slowed and replication initiation events are more frequent. In addition, Mus81 deficient cells fail to recover from exposure to low doses of replication inhibitors and cell viability is dependent on the XPF endonuclease. Despite an increase in replication initiation frequency, cells lacking Mus81 use the same pool of replication origins as Mus81-expressing cells. Therefore, decelerated DNA replication in Mus81 deficient cells does not initiate from cryptic or latent origins not used during normal growth. These results indicate that Mus81 plays a key role in determining the rate of DNA replication without activating a novel group of replication origins.

Differentiating the mineralization dynamics of the originally present and newly synthesized amino acids in soil amended with available carbon and nitrogen substrates

Newly synthesized amino acids are the principle compounds created after inorganic nitrogen (N) is rapidly immobilized into microbial tissues. However, little is known about the mineralization kinetics of these newly synthesized amino acids compared to the amino acids originally present in the soil, and how substrate availability controls their mineralization. With 15N isotope tracing, the newly synthesized (15N-labeled) amino acids can be differentiated from the amino acids originally present (unlabeled) in soil, making it possible to evaluate the mineralization of the newly synthesized amino acids in tandem with the original amino acids. As amino acids can serve as both N and carbon (C) sources for microorganisms, the mineralization dynamics of amino acids may be manipulated by the availability of extraneous C and N. In this study, an aerobic 30-week intermittent leaching experiment was conducted, using glucose as C source and (14NH4)2SO4 as N source, following separate additions to soil. The newly synthesized amino acids were determined by an isotope-based high performance liquid chromatography/mass spectrometry (HPLC/MS). The newly synthesized soil amino acids mineralized faster than the original ones, which indicated more rapid cycling of N in the newly synthesized soil amino acids pool. Glucose addition significantly decreased the mineralization of both the newly synthesized and the original amino acids. However, when inorganic N was abundant, the newly synthesized amino acids decomposed rapidly, and preferentially as a C source and energy, while N addition inhibited the mineralization of the original amino acids in the soil. We conclude that the presence of readily degradable C (e.g. glucose) and inorganic N controls the mineralization of newly synthesized and original amino acid pools in soil differently, which is a crucial mechanism in adjusting the N supply and sequestration processes in soil ecosystems.

Virtual manipulation of topography to test potential pool–riffle maintenance mechanisms

In this study, numerical experimentation with two-dimensional hydraulic modeling of pool–riffle river topography drawing on the testbed data from the classic Keller (1971) study was used to investigate the effect of synthetically manipulating topography on the occurrence and magnitude of velocity and Shields stress reversals in a pool–riffle sequence. Reversals in velocity and shear stress have been used to explore mechanisms of pool–riffle maintenance, while Shields stress (a combined measure of transport capacity and substrate erodibility) is emerging in importance. The original site topography was modeled alongside six altered ones to evaluate the sensitivity of hydraulic reversals to subtle morphology — five incrementally wider pools and a filled pool. The Caamaño (2009) criterion, a simplified geometric threshold for predicting velocity reversals, was applied to each terrain to evaluate its utility. The original pool–riffle topography was just over the threshold for a velocity reversal and well over the threshold for a strong Shields stress reversal. Overall, pool widening caused a predominantly local response, with change to pool hydraulics and no change in section-averaged velocity in the riffle beyond the initial widening of 10%. Filling in the pool significantly increased the magnitude of reversals, whereas expanding it eliminated the occurrence of a reversal in mean velocity, though the Shields stress reversal persisted because of strong differentiation in bed material texture. Using Shields stress as a reversal parameter enabled the quantification of pool modification effects on pool–riffle resiliency. The Caamaño (2009) criterion accurately predicted reversal occurrence for the altered terrains with exaggerated effects, but failed to predict the weak reversal for the original topography. Two-dimensional modeling coupled with previously accepted hydrologic, geomorphic, and engineering analyses is vital in project design and evaluation prior to construction.

Clustering Analysis of Function Call Sequence for Regression Test Case Reduction

Regression test case reduction aims at selecting a representative subset from the original test pool, while retaining the largest possible fault detection capability. Cluster analysis has been proposed and applied for selecting an effective test case subset in regression testing. It groups test cases into clusters based on the similarity of historical execution profiles. In previous studies, historical execution profiles are represented as binary or numeric function coverage vectors. The vector-based similarity approaches only consider which functions or statements are covered and the number of times they are executed. However, the vector-based approaches do not take the relations and sequential information between function calls into account. In this paper, we propose cluster analysis of function call sequences to attempt to improve the fault detection effectiveness of regression testing even further. A test is represented as a function call sequence that includes the relations and sequential information between function calls. The distance between function call sequences is measured not only by the Levenshtein distance but also the Euclidean distance. To assess the effectiveness of our approaches, we designed and conducted experimental studies on five subject programs. The experimental results indicate that our approaches are statistically superior to the approaches based on the similarity of vectors (i.e. binary vectors and numeric vectors), random and greedy function-coverage-based maximization test case reduction techniques in terms of fault detection effectiveness. With respective to the cost-effectiveness, cluster analysis of sequences measured using the Euclidean distance is more effective than using the Levenshtein distance.

A Parameter for Forecasting Tornadoes Associated with Landfalling Tropical Cyclones

Abstract The authors develop a statistical guidance product, the tropical cyclone tornado parameter (TCTP), for forecasting the probability of one or more tornadoes during a 6-h period that are associated with landfalling tropical cyclones affecting the coastal Gulf of Mexico and the southern Atlantic coast. TCTP is designed to aid forecasters in a time-limited environment. TCTP provides a “quick look” at regions where forecasters can then conduct detailed analyses. The pool of potential predictors included tornado reports and tropical cyclone data between 2000 and 2008, as well as storm environmental parameters. The original pool of 28 potential predictors is reduced to six using stepwise regression and logistic regression. These six predictors are 0–3-km wind shear, 0–3-km storm relative helicity, azimuth angle of the tornado report from the tropical cyclone, distance from the cyclone’s center, time of day, and 950–1000-hPa convective available potential energy. Mean Brier scores and Brier skill scores are computed for the entire TCTP-dependent dataset and for corresponding forecasts produced by the Storm Prediction Center (SPC). TCTP then is applied to four individual cyclone cases to qualitatively and quantitatively assess the parameter and compare its performance with SPC forecasts. Results show that TCTP has skill at identifying regions of tornado potential. However, tornadoes in some tropical systems are overpredicted, but underpredicted in others. TCTP 6-h forecast periods provide slightly poorer statistical performance than the 1-day tornado probability forecasts from SPC, probably because the SPC product includes forecaster guidance and because their forecasts are valid for longer periods (24 h).

Initial establishment of the cognitive questionnaire about blood pressure monitoring in critical care nurses

Objective To develop the suitable for our country , high reliability and validity of cognitive questionnaire about blood pressure monitoring in critical care nurses （ CCNs ） in order to provide the help for understanding the status of blood pressure monitoring in CCNs in our country .Methods The original entry pool was preliminarily established according the international assessment scale of clinical outcome in patients through the structured decision-making including the McGhee-items, references from databases and experts ’ modification.70 CCNs and 120 undergraduate nursing students with one ICU at least were surveyed by the preliminary questionnaire .The entries were screened again through the investigation of entry distribution , dispersion, t test, resolution coefficient and the discussion in core group again , and the reliability and validity of internal consistency were evaluated .Results The original entry pool was 35 items, and the items of experts ’ modification was 3;5 items were deleted through the item selection combined with the discussion in core group . The items of the remaining entries in three dimensions including the physiology of blood pressure monitoring , technical aspects of monitoring , wave distinguish and data interpretation were respectively 12, 8, 10 items.The total Cronbach ’ s αcoefficient of questionnaire was 0.783, and the Cronbach ’ s αcoefficient of different dimensions were respectively 0.822, 0.793 and 0.734.Six extracting factors explained 82.57% of the total variance, and the loads of the corresponding dimensions were more than 0.55.Conclusions The initial questionnaire which is consisted of items after selection can comprehensively reflect the connotation of blood pressure monitoring , and can be used for investigating the cognitive level of blood pressure monitoring in CCNs in our country due to high validity and reliability . Key words: Blood pressure monitoring; Entry pool; Item selection; Critical care nurses