Search results for: specificity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 556

Search results for: specificity

556 Cone Contrast Sensitivity of Normal Trichromats and Those with Red-Green Dichromats

Authors: Tatsuya Iizuka, Takushi Kawamorita, Tomoya Handa, Hitoshi Ishikawa

Abstract:

We report normative cone contrast sensitivity values and sensitivity and specificity values for a computer-based color vision test, the cone contrast test-HD (CCT-HD). The participants included 50 phakic eyes with normal color vision (NCV) and 20 dichromatic eyes (ten with protanopia and ten with deuteranopia). The CCT-HD was used to measure L, M, and S-CCT-HD scores (color vision deficiency, L-, M-cone logCS≦1.65, S-cone logCS≦0.425) to investigate the sensitivity and specificity of CCT-HD based on anomalous-type diagnosis with animalscope. The mean ± standard error L-, M-, S-cone logCS for protanopia were 0.90±0.04, 1.65±0.03, and 0.63±0.02, respectively; for deuteranopia 1.74±0.03, 1.31±0.03, and 0.61±0.06, respectively; and for age-matched NCV were 1.89±0.04, 1.84±0.04, and 0.60±0.03, respectively, with significant differences for each group except for S-CCT-HD (Bonferroni corrected α = 0.0167, p < 0.0167). The sensitivity and specificity of CCT-HD were 100% for protan and deutan in diagnosing abnormal types from 20 to 64 years of age, but the specificity decreased to 65% for protan and 55% for deutan in older persons > 65. CCT-HD is comparable to the diagnostic performance of the anomalous type in the anomaloscope for the 20-64-year-old age group. However, the results should be interpreted cautiously in those ≥ 65 years. They are more susceptible to acquired color vision deficiencies due to the yellowing of the crystalline lens and other factors.

Keywords: cone contrast test HD, color vision test, congenital color vision deficiency, red-green dichromacy, cone contrast sensitivity

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555 Evaluation of 18F Fluorodeoxyglucose Positron Emission Tomography, MRI, and Ultrasound in the Assessment of Axillary Lymph Node Metastases in Patients with Early Stage Breast Cancer

Authors: Wooseok Byon, Eunyoung Kim, Junseong Kwon, Byung Joo Song, Chan Heun Park

Abstract:

Purpose: 18F Fluorodeoxyglucose Positron Emission Tomography (FDG-PET) is a noninvasive imaging modality that can identify nodal metastases in women with primary breast cancer. The aim of this study was to compare the accuracy of FDG-PET with MRI and sonography scanning to determine axillary lymph node status in patients with breast cancer undergoing sentinel lymph node biopsy or axillary lymph node dissection. Patients and Methods: Between January and December 2012, ninety-nine patients with breast cancer and clinically negative axillary nodes were evaluated. All patients underwent FDG-PET, MRI, ultrasound followed by sentinel lymph node biopsy (SLNB) or axillary lymph node dissection (ALND). Results: Using axillary lymph node assessment as the gold standard, the sensitivity and specificity of FDG-PET were 51.4% (95% CI, 41.3% to 65.6%) and 92.2% (95% CI, 82.7% to 97.4%) respectively. The sensitivity and specificity of MRI and ultrasound were 57.1% (95% CI, 39.4% to 73.7%), 67.2% (95% CI, 54.3% to 78.4%) and 42.86% (95% CI, 26.3% to 60.7%), 92.2% (95% CI, 82.7% to 97.4%). Stratification according to hormone receptor status showed an increase in specificity when negative (FDG-PET: 42.3% to 77.8%, MRI 50% to 77.8%, ultrasound 34.6% to 66.7%). Also, positive HER2 status was associated with an increase in specificity (FDG-PET: 42.9% to 85.7%, MRI 50% to 85.7%, ultrasound 35.7% to 71.4%). Conclusions: The sensitivity and specificity of FDG-PET compared with MRI and ultrasound was high. However, FDG-PET is not sufficiently accurate to appropriately identify lymph node metastases. This study suggests that FDG-PET scanning cannot replace histologic staging in early-stage breast cancer, but might have a role in evaluating axillary lymph node status in hormone receptor negative or HER-2 overexpressing subtypes.

Keywords: axillary lymph node metastasis, FDG-PET, MRI, ultrasound

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554 CRISPR-DT: Designing gRNAs for the CRISPR-Cpf1 System with Improved Target Efficiency and Specificity

Authors: Houxiang Zhu, Chun Liang

Abstract:

The CRISPR-Cpf1 system has been successfully applied in genome editing. However, target efficiency of the CRISPR-Cpf1 system varies among different gRNA sequences. The published CRISPR-Cpf1 gRNA data was reanalyzed. Many sequences and structural features of gRNAs (e.g., the position-specific nucleotide composition, position-nonspecific nucleotide composition, GC content, minimum free energy, and melting temperature) correlated with target efficiency were found. Using machine learning technology, a support vector machine (SVM) model was created to predict target efficiency for any given gRNAs. The first web service application, CRISPR-DT (CRISPR DNA Targeting), has been developed to help users design optimal gRNAs for the CRISPR-Cpf1 system by considering both target efficiency and specificity. CRISPR-DT will empower researchers in genome editing.

Keywords: CRISPR-Cpf1, genome editing, target efficiency, target specificity

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553 Systematic Identification and Quantification of Substrate Specificity Determinants in Human Protein Kinases

Authors: Manuel A. Alonso-Tarajano, Roberto Mosca, Patrick Aloy

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Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase– pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed- specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Keywords: kinase, phosphorylation, substrate specificity, adaptors, scaffolds, cellular colocalization

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552 Identifying the True Extend of Glioblastoma Based on Preoperative FLAIR Images

Authors: B. Shukir, L. Szivos, D. Kis, P. Barzo

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Glioblastoma is the most malignant brain tumor. In general, the survival rate varies between (14-18) months. Glioblastoma consists a solid and infiltrative part. The standard therapeutic management of glioblastoma is maximum safe resection followed by chemo-radiotherapy. It’s hypothesized that the pretumoral hyperintense region in fluid attenuated inversion recovery (FLAIR) images includes both vasogenic edema and infiltrated tumor cells. In our study, we aimed to define the sensitivity and specificity of hyperintense FLAIR images preoperatively to examine how well it can define the true extent of glioblastoma. (16) glioblastoma patients included in this study. Hyperintense FLAIR region were delineated preoperatively as tumor mask. The infiltrative part of glioblastoma considered the regions where the tumor recurred on the follow up MRI. The recurrence on the CE-T1 images was marked as the recurrence masks. According to (AAL3) and (JHU white matter labels) atlas, the brain divided into cortical and subcortical regions respectively. For calculating specificity and sensitivity, the FLAIR and the recurrence masks overlapped counting how many regions affected by both . The average sensitivity and specificity was 83% and 85% respectively. Individually, the sensitivity and specificity varied between (31-100)%, and (100-58)% respectively. These results suggest that despite FLAIR being as an effective radiologic imaging tool its prognostic value remains controversial and probabilistic tractography remain more reliable available method for identifying the true extent of glioblastoma.

Keywords: brain tumors, glioblastoma, MRI, FLAIR

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551 Novel Ultrasensitive Point of Care Device for Diagnosis of Human Schistosomiasis Mansoni

Authors: Ibrahim Aly, Waleed Elawamy, Hanan Taher, Amira Matar

Abstract:

Schistosomiasis is infection with blood flukes of the genus Schistosoma, which are acquired trans-cutaneously by swimming or wading in contaminated freshwater. The present study was proposed to produce ultra-sensitive, field-friendly high-throughput rapid immunochromatography diagnostic device for accurate detection of asymptomatic parasite carriers in schistosomiasis pre-elimination settings.For assessing diagnostic potential of rapid device, 50 blood samples from patients with schistosomiasis mansoni, 29 other proven parasitic diseases and 25 blood samples as negative control were from healthy individuals were used. The sensitivity of Quantitative antigen-capture nano-ELISAwas 82 %, and specificity was 87.1 %, where the sensitivity of Nano Dot- ELISA was 86 % and specificity was 90.7 %. The sensitivity of diagnostic device was 78 % and specificity was 85.2 %, with PPV and NPV of 86.2 % and 83.1 %, respectively.The Point of care device resulted in a good performance for the diagnosis of low-intensity infections, it was able to identify 19 out of 25 (76 %) individuals with ⩽7 eggs, 10 out of 14 individuals (71.4 %) with 11–99 eggs and 100 % of individuals with 100–399 eggs.

Keywords: schistosomiasis, immunochromatography, naon-dot-ELISa, diagnostis device

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550 Evaluation of the Benefit of Anti-Endomysial IgA and Anti-Tissue Transglutaminase IgA Antibodies for the Diagnosis of Coeliac Disease in a University Hospital, 2010-2016

Authors: Recep Keşli, Onur Türkyılmaz, Hayriye Tokay, Kasım Demir

Abstract:

Objective: Coeliac disease (CD) is a primary small intestine disorder caused by high sensitivity to gluten which is present in the crops, characterized by inflammation in the small intestine mucosa. The goal of this study was to determine and to compare the sensitivity and specificity values of anti-endomysial IgA (EMA IgA) (IFA) and anti-tissue transglutaminase IgA (anti-tTG IgA) (ELISA) antibodies in the diagnosis of patients suspected with the CD. Methods: One thousand two hundred seventy three patients, who have applied to gastroenterology and pediatric disease polyclinics of Afyon Kocatepe University ANS Research and Practice Hospital were included into the study between 23.09.2010 and 30.05.2016. Sera samples were investigated by immunofluorescence method for EMA positiveness (Euroimmun, Luebeck, Germany). In order to determine quantitative value of Anti-tTG IgA (EIA) (Orgentec Mainz, Germany) fully automated ELISA device (Alisei, Seac, Firenze, Italy) were used. Results: Out of 1273 patients, 160 were diagnosed with coeliac disease according to ESPGHAN 2012 diagnosis criteria. Out of 160 CD patients, 120 were female, 40 were male. The EMA specificity and sensitivity were calculated as 98% and 80% respectively. Specificity and sensitivity of Anti-tTG IgA were determined as 99% and 96% respectively. Conclusion: The specificity of EMA for CD was excellent because all EMA-positive patients (n = 144) were diagnosed with CD. The presence of human anti-tTG IgA was found as a reliable marker for diagnosis and follow-up the CD. Diagnosis of CD should be established on both the clinical and serologic profiles together.

Keywords: anti-endomysial antibody, anti-tTG IgA, coeliac disease, immunofluorescence assay (IFA)

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549 Developing HRCT Criterion to Predict the Risk of Pulmonary Tuberculosis

Authors: Vandna Raghuvanshi, Vikrant Thakur, Anupam Jhobta

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Objective: To design HRCT criterion to forecast the threat of pulmonary tuberculosis. Material and methods: This was a prospective study of 69 patients with clinical suspicion of pulmonary tuberculosis. We studied their medical characteristics, numerous separate HRCT-results, and a combination of HRCT findings to foresee the danger for PTB by utilizing univariate and multivariate investigation. Temporary HRCT diagnostic criteria were planned in view of these outcomes to find out the risk of PTB and tested these criteria on our patients. Results: The results of HRCT chest were analyzed, and Rank was given from 1 to 4 according to the HRCT chest findings. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated. Rank 1: Highly suspected PTB. Rank 2: Probable PTB Rank 3: Nonspecific or difficult to differentiate from other diseases Rank 4: Other suspected diseases • Rank 1 (Highly suspected TB) was present in 22 (31.9%) patients, all of them finally diagnosed to have pulmonary tuberculosis. The sensitivity, specificity, and negative likelihood ratio for RANK 1 on HRCT chest was 53.6%, 100%, and 0.43, respectively. • Rank 2 (Probable TB) was present in 13 patients, out of which 12 were tubercular, and 1 was non-tubercular. • The sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio of the combination of Rank 1 and Rank 2 was 82.9%, 96.4%, 23.22, and 0.18, respectively. • Rank 3 (Non-specific TB) was present in 25 patients, and out of these, 7 were tubercular, and 18 were non-tubercular. • When all these 3 ranks were considered together, the sensitivity approached 100% however, the specificity reduced to 35.7%. The positive likelihood ratio and negative likelihood ratio were 1.56 and 0, respectively. • Rank 4 (Other specific findings) was given to 9 patients, and all of these were non-tubercular. Conclusion: HRCT is useful in selecting individuals with greater chances of pulmonary tuberculosis.

Keywords: pulmonary, tuberculosis, multivariate, HRCT

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548 The Determinants of Co-Production for Value Co-Creation: Quadratic Effects

Authors: Li-Wei Wu, Chung-Yu Wang

Abstract:

Recently, interest has been generated in the search for a new reference framework for value creation that is centered on the co-creation process. Co-creation implies cooperative value creation between service firms and customers and requires the building of experiences as well as the resolution of problems through the combined effort of the parties in the relationship. For customers, values are always co-created through their participation in services. Customers can ultimately determine the value of the service in use. This new approach emphasizes that a customer’s participation in the service process is considered indispensable to value co-creation. An important feature of service in the context of exchange is co-production, which implies that a certain amount of participation is needed from customers to co-produce a service and hence co-create value. Co-production no doubt helps customers better understand and take charge of their own roles in the service process. Thus, this proposal is to encourage co-production, thus facilitating value co-creation of that is reflected in both customers and service firms. Four determinants of co-production are identified in this study, namely, commitment, trust, asset specificity, and decision-making uncertainty. Commitment is an essential dimension that directly results in successful cooperative behaviors. Trust helps establish a relational environment that is fundamental to cross-border cooperation. Asset specificity motivates co-production because this determinant may enhance return on asset investment. Decision-making uncertainty prompts customers to collaborate with service firms in making decisions. In other words, customers adjust their roles and are increasingly engaged in co-production when commitment, trust, asset specificity, and decision-making uncertainty are enhanced. Although studies have examined the preceding effects, to our best knowledge, none has empirically examined the simultaneous effects of all the curvilinear relationships in a single study. When these determinants are excessive, however, customers will not engage in co-production process. In brief, we suggest that the relationships of commitment, trust, asset specificity, and decision-making uncertainty with co-production are curvilinear or are inverse U-shaped. These new forms of curvilinear relationships have not been identified in existing literature on co-production; therefore, they complement extant linear approaches. Most importantly, we aim to consider both the bright and the dark sides of the determinants of co-production.

Keywords: co-production, commitment, trust, asset specificity, decision-making uncertainty

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547 Sensitivity and Specificity of Clinical Testing for Digital Nerve Injury

Authors: Guy Rubin, Ravit Shay, Nimrod Rozen

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The accuracy of a diagnostic test used to classify a patient as having disease or being disease-free is a valuable piece of information to be used by the physician when making treatment decisions. Finger laceration, suspected to have nerve injury is a challenging decision for the treating surgeon. The purpose of this study was to evaluate the sensitivity, specificity and predictive values of six clinical tests in the diagnosis of digital nerve injury. The six clinical tests included light touch, pin prick, static and dynamic 2-point discrimination, Semmes Weinstein monofilament and wrinkle test. Data comparing pre-surgery examination with post-surgery results of 42 patients with 52 digital nerve injury was evaluated. The subjective examinations, light touch, pin prick, static and dynamic 2-point discrimination and Semmes-Weinstein monofilament were not sensitive (57.6, 69.7, 42.4, 40 and 66.8% respectively) and specific (36.8, 36.8, 47.4, 42.1 and 31.6% respectively). Wrinkle test, the only objective examination, was the most sensitive (78.1%) and specific (55.6%). This result gives no pre-operative examination the ability to predict the result of explorative surgery.

Keywords: digital nerve, injury, nerve examination, Semmes-Weinstein monofilamen, sensitivity, specificity, two point discrimination, wrinkle test

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546 Comparison of Sensitivity and Specificity of Pap Smear and Polymerase Chain Reaction Methods for Detection of Human Papillomavirus: A Review of Literature

Authors: M. Malekian, M. E. Heydari, M. Irani Estyar

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Human papillomavirus (HPV) is one of the most common sexually transmitted infection, which may lead to cervical cancer as the main cause of it. With early diagnosis and treatment in health care services, cervical cancer and its complications are considered to be preventable. This study was aimed to compare the efficiency, sensitivity, and specificity of Pap smear and polymerase chain reaction (PCR) in detecting HPV. A literature search was performed in Google Scholar, PubMed and SID databases using the keywords 'human papillomavirus', 'pap smear' and 'polymerase change reaction' to identify studies comparing Pap smear and PCR methods for the detection. No restrictions were considered.10 studies were included in this review. All samples that were positive by pop smear were also positive by PCR. However, there were positive samples detected by PCR which was negative by pop smear and in all studies, many positive samples were missed by pop smear technique. Although The Pap smear had high specificity, PCR based HPV detection was more sensitive method and had the highest sensitivity. In order to promote the quality of detection and high achievement of the maximum results, PCR diagnostic methods in addition to the Pap smear are needed and Pap smear method should be combined with PCR techniques according to the high error rate of Pap smear in detection.

Keywords: human papillomavirus, cervical cancer, pap smear, polymerase chain reaction

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545 A Comparison Study of Different Methods Used in the Detection of Giardia lamblia on Fecal Specimen of Children

Authors: Muhammad Farooq Baig

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Objective: The purpose of this study was to compare results obtained using a single fecal specimen for O&P examination, direct immunofluorescence assay (DFA), and two conventional staining methods. Design: Hundred and fifty children fecal specimens were collected and examined by each method. The O&P and the DFA were used as the reference method. Setting: The study was performed at the laboratory in the Basic Medical Science Institute JPMC Karachi. Patients or Other Participants: The fecal specimens were collected from children with a suspected Giardia lamblia infection. Main Outcome Measures: The amount of agreement and disagreement between methods.1) Presence of giardiasis in our population. 2) The sensitivity and specificity of each method. Results: There was 45(30%) positive 105 (70%) negative on DFA, 41 (27.4%) positive 109 (72.6%) negative on iodine and 34 (22.6%) positive 116(77.4%) on saline method. The sensitivity and specificity of DFA in comparision to iodine were 92.2%, 92.7% respectively. The sensitivity and specificity of DFA in comparisoin to saline method were 91.2%, 87.9% respectively. The sensitivity of iodine method and saline method in compariosn to DFA were 82.2%, 68.8% respectively. There is mark diffrence in sensitivity of DFA to conventional method. Conclusion: The study supported findings of other investigators who concluded that DFA method have the greater sensitivity. The immunologic methods were more efficient and quicker than the conventional O&P method.

Keywords: direct immunofluorescence assay (DFA), ova and parasite (O&P), Giardia lamblia, children, medical science

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544 The Specificity of Employee Development in Polish Small Enterprises

Authors: E. Rak

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The aim of the paper is to identify some of the specific characteristics of employee development, as observed in the practice of small enterprises in Poland. Results suggest that a sizeable percentage of employers are not interested in improving the development of their employee base. This aspect is often perceived as insignificant. In addition, many employers have no theoretical or practical knowledge of employee development methods. Lack of sufficient financial support is reported as third on the list of the most important barriers to employee development. Employees, on the other hand, typically offload the responsibility of initiating this type of activities onto the employer. Employee development plans are typically flexible and accommodating. The original value offered by this research comes in the form of a detailed characteristics of employee development in small enterprises, accompanied by identification of specificity of human resource development in Polish companies.

Keywords: employee development, human resources development, small enterprises, trainings

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543 Biopsy or Biomarkers: Which Is the Sample of Choice in Assessment of Liver Fibrosis?

Authors: S. H. Atef, N. H. Mahmoud, S. Abdrahman, A. Fattoh

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Background: The aim of the study is to assess the diagnostic value of fibrotest and hyaluronic acid in discriminate between insignificant and significant fibrosis. Also, to find out if these parameters could replace liver biopsy which is currently used for selection of chronic hepatitis C patients eligible for antiviral therapy. Study design: This study was conducted on 52 patients with HCV RNA detected by polymerase chain reaction (PCR) who had undergone liver biopsy and attending the internal medicine clinic at Ain Shams University Hospital. Liver fibrosis was evaluated according to the METAVIR scoring system on a scale of F0 to F4. Biochemical markers assessed were: alpha-2 macroglobulin (α2-MG), apolipoprotein A1 (Apo-A1), haptoglobin, gamma-glutamyl transferase (GGT), total bilirubin (TB) and hyaluronic acid (HA). The fibrotest score was computed after adjusting for age and gender. Predictive values and ROC curves were used to assess the accuracy of fibrotest and HA results. Results: For fibrotest, the observed area under curve for the discrimination between minimal or no fibrosis (F0-F1) and significant fibrosis (F2-F4) was 0.6736 for cutoff value 0.19 with sensitivity of 84.2% and specificity of 85.7%. For HA, the sensitivity was 89.5% and specificity was 85.7% and area under curve was 0.540 at the best cutoff value 71 mg/dL. Multi-use of both parameters, HA at 71 mg/dL with fibrotest score at 0.22 give a sensitivity 89.5%, specificity 100 and efficacy 92.3% (AUC 0.895). Conclusion: The use of both fibrotest score and HA could be as alternative to biopsy in most patients with chronic hepaitis C putting in consideration some limitations of the proposed markers in evaluating liver fibrosis.

Keywords: fibrotest, liver fibrosis, HCV RNA, biochemical markers

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542 Increase in Specificity of MicroRNA Detection by RT-qPCR Assay Using a Specific Extension Sequence

Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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We describe an innovative method for highly specific detection of miRNAs using a specially modified method of poly(A) adaptor RT-qPCR. We use uniquely designed specific extension sequence, which plays important role in providing an opportunity to affect high specificity of miRNA detection. This method involves two steps of reactions as like previously reported and which are poly(A) tailing and reverse-transcription followed by real-time PCR. Firstly, miRNAs are extended by a poly(A) tailing reaction and then converted into cDNA. Here, we remarkably reduced the reaction time by the application of short length of poly(T) adaptor. Next, cDNA is hybridized to the 3’-end of a specific extension sequence which contains miRNA sequence and results in producing a novel PCR template. Thereafter, the SYBR Green-based RT-qPCR progresses with a universal poly(T) adaptor forward primer and a universal reverse primer. The target miRNA, miR-106b in human brain total RNA, could be detected quantitatively in the range of seven orders of magnitude, which demonstrate that the assay displays a dynamic range of at least 7 logs. In addition, the better specificity of this novel extension-based assay against well known poly(A) tailing method for miRNA detection was confirmed by melt curve analysis of real-time PCR product, clear gel electrophoresis and sequence chromatogram images of amplified DNAs.

Keywords: microRNA(miRNA), specific extension sequence, RT-qPCR, poly(A) tailing assay, reverse transcription

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541 The Use of Venous Glucose, Serum Lactate and Base Deficit as Biochemical Predictors of Mortality in Polytraumatized Patients: Acomparative with Trauma and Injury Severity Score and Acute Physiology and Chronic Health Evalution IV

Authors: Osama Moustafa Zayed

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Aim of the work: To evaluate the effectiveness of venous glucose, levels of serum lactate and base deficit in polytraumatized patients as simple parameters to predict the mortality in these patients. Compared to the predictive value of Trauma and injury severity (TRISS) and Acute Physiology And Chronic Health Evaluation IV (APACHE IV). Introduction: Trauma is a serious global health problem, accounting for approximately one in 10 deaths worldwide. Trauma accounts for 5 million deaths per year. Prediction of mortality in trauma patients is an important part of trauma care. Several trauma scores have been devised to predict injury severity and risk of mortality. The trauma and injury severity score (TRISS) was most common used. Regardless of the accuracy of trauma scores, is based on an anatomical description of every injury and cannot be assigned to the patients until a full diagnostic procedure has been performed. So we hypothesized that alterations in admission glucose, lactate levels and base deficit would be an early and easy rapid predictor of mortality. Patient and Method: a comparative cross-sectional study. 282 Polytraumatized patients attended to the Emergency Department(ED) of the Suez Canal university Hospital constituted. The period from 1/1/2012 to 1/4/2013 was included. Results: We found that the best cut off value of TRISS probability of survival score for prediction of mortality among poly-traumatized patients is = 90, with 77% sensitivity and 89% specificity using area under the ROC curve (0.89) at (95%CI). APACHE IV demonstrated 67% sensitivity and 95% specificity at 95% CI at cut off point 99. The best cutoff value of Random Blood Sugar (RBS) for prediction of mortality was>140 mg/dl, with 89%, sensitivity, 49% specificity. The best cut off value of base deficit for prediction of mortality was less than -5.6 with 64% sensitivity, 93% specificity. The best cutoff point of lactate for prediction of mortality was > 2.6 mmol/L with 92%, sensitivity, 42% specificity. Conclusion: According to our results from all evaluated predictors of mortality (laboratory and scores) and mortality based on the estimated cutoff values using ROC curves analysis, the highest risk of mortality was found using a cutoff value of 90 in TRISS score while with laboratory parameters the highest risk of mortality was with serum lactate > 2.6 . Although that all of the three parameter are accurate in predicting mortality in poly-traumatized patients and near with each other, as in serum lactate the area under the curve 0.82, in BD 0.79 and 0.77 in RBS.

Keywords: APACHE IV, emergency department, polytraumatized patients, serum lactate

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540 Comparative Study of the Sensitivity of Two Freshwater Gastropods, Lymnaea Stagnalis and Planorbarius Corneus, to Silver Nanoparticles: Bioaccumulation and Toxicity

Authors: Ting Wang, Pierre Marle, Vera I. Slaveykova, Kristin Schirmer, Wei Liu

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Metal-based nanoparticles (NPs) are considered detrimental to aquatic organisms due to their potential accumulation. However, little is known about the mechanisms underlying these effects and their species-specificity. Here, we used stable silver (Ag) NPs (20 nm, from 10 to 500 μg/L) with a low dissolution rate (≤2.4%) to study the bioaccumulation and biological impacts in two freshwater gastropods: Lymnaea stagnalis and Planorbarius corneus. No mortality was detected during the experiments. Ag bioaccumulation showed a dose-related increase with an enhanced concentration in both species after 7d exposure. L. stagnalis displayed a higher accumulation for AgNPs than P. corneus (e.g., up to 18- and 15-fold in hepatopancreas and hemolymph, respectively), which could be due to the more active L. stagnalis having greater contact with suspended AgNPs. Furthermore, the hepatopancreas and stomach were preferred organs for bioaccumulation compared to the kidney, mantle and foot. Regarding biological responses, the hemolymph rather than hepatopancreas appeared more susceptible to oxidative stress elicited by AgNPs, as shown by significantly increasing lipid peroxidation (i.e., formation of malondialdehyde). Neurotoxicity was detected in L. stagnalis when exposed to high concentrations (500 μg/L). Comparison with impacts elicited by dissolved Ag revealed that the effects observed on AgNPs exposure were mainly attributable to NPs. These results highlighted the relationship between the physiological traits, bioaccumulation, and toxicity responses of these two species to AgNPs and demonstrated the necessity of species-specificity considerations when assessing the toxicity of NPs.

Keywords: nanotoxicity, freshwater gastropods, species-specificity, metals, physiological traits

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539 Diagnostic Accuracy of the Tuberculin Skin Test for Tuberculosis Diagnosis: Interest of Using ROC Curve and Fagan’s Nomogram

Authors: Nouira Mariem, Ben Rayana Hazem, Ennigrou Samir

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Background and aim: During the past decade, the frequency of extrapulmonary forms of tuberculosis has increased. These forms are under-diagnosed using conventional tests. The aim of this study was to evaluate the performance of the Tuberculin Skin Test (TST) for the diagnosis of tuberculosis, using the ROC curve and Fagan’s Nomogram methodology. Methods: This was a case-control, multicenter study in 11 anti-tuberculosis centers in Tunisia, during the period from June to November2014. The cases were adults aged between 18 and 55 years with confirmed tuberculosis. Controls were free from tuberculosis. A data collection sheet was filled out and a TST was performed for each participant. Diagnostic accuracy measures of TST were estimated using ROC curve and Area Under Curve to estimate sensitivity and specificity of a determined cut-off point. Fagan’s nomogram was used to estimate its predictive values. Results: Overall, 1053 patients were enrolled, composed of 339 cases (sex-ratio (M/F)=0.87) and 714 controls (sex-ratio (M/F)=0.99). The mean age was 38.3±11.8 years for cases and 33.6±11 years for controls. The mean diameter of the TST induration was significantly higher among cases than controls (13.7mm vs.6.2mm;p=10-6). Area Under Curve was 0.789 [95% CI: 0.758-0.819; p=0.01], corresponding to a moderate discriminating power for this test. The most discriminative cut-off value of the TST, which were associated with the best sensitivity (73.7%) and specificity (76.6%) couple was about 11 mm with a Youden index of 0.503. Positive and Negative predictive values were 3.11% and 99.52%, respectively. Conclusion: In view of these results, we can conclude that the TST can be used for tuberculosis diagnosis with a good sensitivity and specificity. However, the skin induration measurement and its interpretation is operator dependent and remains difficult and subjective. The combination of the TST with another test such as the Quantiferon test would be a good alternative.

Keywords: tuberculosis, tuberculin skin test, ROC curve, cut-off

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538 Comparison of the Classification of Cystic Renal Lesions Using the Bosniak Classification System with Contrast Enhanced Ultrasound and Magnetic Resonance Imaging to Computed Tomography: A Prospective Study

Authors: Dechen Tshering Vogel, Johannes T. Heverhagen, Bernard Kiss, Spyridon Arampatzis

Abstract:

In addition to computed tomography (CT), contrast enhanced ultrasound (CEUS), and magnetic resonance imaging (MRI) are being increasingly used for imaging of renal lesions. The aim of this prospective study was to compare the classification of complex cystic renal lesions using the Bosniak classification with CEUS and MRI to CT. Forty-eight patients with 65 cystic renal lesions were included in this study. All participants signed written informed consent. The agreement between the Bosniak classifications of complex renal lesions ( ≥ BII-F) on CEUS and MRI were compared to that of CT and were tested using Cohen’s Kappa. Sensitivity, specificity, positive and negative predictive values (PPV/NPV) and the accuracy of CEUS and MRI compared to CT in the detection of complex renal lesions were calculated. Twenty-nine (45%) out of 65 cystic renal lesions were classified as complex using CT. The agreement between CEUS and CT in the classification of complex cysts was fair (agreement 50.8%, Kappa 0.31), and was excellent between MRI and CT (agreement 93.9%, Kappa 0.88). Compared to CT, MRI had a sensitivity of 96.6%, specificity of 91.7%, a PPV of 54.7%, and an NPV of 54.7% with an accuracy of 63.1%. The corresponding values for CEUS were sensitivity 100.0%, specificity 33.3%, PPV 90.3%, and NPV 97.1% with an accuracy 93.8%. The classification of complex renal cysts based on MRI and CT scans correlated well, and MRI can be used instead of CT for this purpose. CEUS can exclude complex lesions, but due to higher sensitivity, cystic lesions tend to be upgraded. However, it is useful for initial imaging, for follow up of lesions and in those patients with contraindications to CT and MRI.

Keywords: Bosniak classification, computed tomography, contrast enhanced ultrasound, cystic renal lesions, magnetic resonance imaging

Procedia PDF Downloads 110
537 New Kinetic Approach to the Enzymatic Hydrolysis of Proteins: A Case of Thermolysin-Catalyzed Albumin

Authors: Anna Trusek-Holownia, Andrzej Noworyta

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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides

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536 Clinical Trial of VEUPLEXᵀᴹ TBI Assay to Help Diagnose Traumatic Brain Injury by Quantifying Glial Fibrillary Acidic Protein and Ubiquitin Carboxy-Terminal Hydrolase L1 in the Serum of Patients Suspected of Mild TBI by Fluorescence Immunoassay

Authors: Moon Jung Kim, Guil Rhim

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The clinical sensitivity of the “VEUPLEXTM TBI assay”, a clinical trial medical device, in mild traumatic brain injury was 28.6% (95% CI, 19.7%-37.5%), and the clinical specificity was 94.0% (95% CI, 89.3%). -98.7%). In addition, when the results analyzed by marker were put together, the sensitivity was higher when interpreting the two tests together than the two tests, UCHL1 and GFAP alone. Additionally, when sensitivity and specificity were analyzed based on CT results for the mild traumatic brain injury patient group, the clinical sensitivity for 2 CT-positive cases was 50.0% (95% CI: 1.3%-98.7%), and 19 CT-negative cases. The clinical specificity for cases was 68.4% (95% CI: 43.5% - 87.4%). Since the low clinical sensitivity for the two CT-positive cases was not statistically significant due to the small number of samples analyzed, it was judged necessary to secure and analyze more samples in the future. Regarding the clinical specificity analysis results for 19 CT-negative cases, there were a large number of patients who were actually clinically diagnosed with mild traumatic brain injury but actually received a CT-negative result, and about 31.6% of them showed abnormal results on VEUPLEXTM TBI assay. Although traumatic brain injury could not be detected in 31.6% of the CT scans, the possibility of actually suffering a mild brain injury could not be ruled out, so it was judged that this could be confirmed through follow-up observation of the patient. In addition, among patients with mild traumatic brain injury, CT examinations were not performed in many cases because the symptoms were very mild, but among these patients, about 25% or more showed abnormal results in the VEUPLEXTM TBI assay. In fact, no damage is observed with the naked eye immediately after traumatic brain injury, and traumatic brain injury is not observed even on CT. But in some cases, brain hemorrhage may occur (delayed cerebral hemorrhage) after a certain period of time, so the patients who did show abnormal results on VEUPLEXTM TBI assay should be followed up for the delayed cerebral hemorrhage. In conclusion, it was judged that it was difficult to judge mild traumatic brain injury with the VEUPLEXTM TBI assay only through clinical findings without CT results, that is, based on the GCS value. Even in the case of CT, it does not detect all mild traumatic brain injury, so it is difficult to necessarily judge that there is no traumatic brain injury, even if there is no evidence of traumatic brain injury in CT. And in the long term, more patients should be included to evaluate the usefulness of the VEUPLEXTM TBI assay in the detection of microscopic traumatic brain injuries without using CT.

Keywords: brain injury, traumatic brain injury, GFAP, UCHL1

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535 Employee Assessment Systems in the Structures of Corporate Groups

Authors: D. Bąk-Grabowska, K. Grzesik, A. Iwanicka, A. Jagoda

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The process of human resources management in the structures of corporate groups demonstrates certain specificity, resulting from the division of decision-making and executive competencies, which occurs within these structures between a parent company and its subsidiaries. The subprocess of employee assessment is considered crucial, since it provides information for the implementation of personnel function. The empirical studies conducted in corporate groups, within which at least one company is located in Poland, confirmed the critical significance of employee assessment systems in the process of human resources management in corporate groups. Parent companies, most often, retain their decision-making authority within the framework of the discussed process and introduce uniform employee assessment and personnel controlling systems to subsidiary companies. However, the instruments for employee assessment applied in corporate groups do not present such specificity.

Keywords: corporate groups, employee periodical assessment system, holding, human resources management

Procedia PDF Downloads 388
534 Performance of the Aptima® HIV-1 Quant Dx Assay on the Panther System

Authors: Siobhan O’Shea, Sangeetha Vijaysri Nair, Hee Cheol Kim, Charles Thomas Nugent, Cheuk Yan William Tong, Sam Douthwaite, Andrew Worlock

Abstract:

The Aptima® HIV-1 Quant Dx Assay is a fully automated assay on the Panther system. It is based on Transcription-Mediated Amplification and real time detection technologies. This assay is intended for monitoring HIV-1 viral load in plasma specimens and for the detection of HIV-1 in plasma and serum specimens. Nine-hundred and seventy nine specimens selected at random from routine testing at St Thomas’ Hospital, London were anonymised and used to compare the performance of the Aptima HIV-1 Quant Dx assay and Roche COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test, v2.0. Two-hundred and thirty four specimens gave quantitative HIV-1 viral load results in both assays. The quantitative results reported by the Aptima Assay were comparable those reported by the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, v2.0 with a linear regression slope of 1.04 and an intercept on -0.097. The Aptima assay detected HIV-1 in more samples than the Roche assay. This was not due to lack of specificity of the Aptima assay because this assay gave 99.83% specificity on testing plasma specimens from 600 HIV-1 negative individuals. To understand the reason for this higher detection rate a side-by-side comparison of low level panels made from the HIV-1 3rd international standard (NIBSC10/152) and clinical samples of various subtypes were tested in both assays. The Aptima assay was more sensitive than the Roche assay. The good sensitivity, specificity and agreement with other commercial assays make the HIV-1 Quant Dx Assay appropriate for both viral load monitoring and detection of HIV-1 infections.

Keywords: HIV viral load, Aptima, Roche, Panther system

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533 Analytical Validity Of A Tech Transfer Solution To Internalize Genetic Testing

Authors: Lesley Northrop, Justin DeGrazia, Jessica Greenwood

Abstract:

ASPIRA Labs now offers an en-suit and ready-to-implement technology transfer solution to enable labs and hospitals that lack the resources to build it themselves to offer in-house genetic testing. This unique platform employs a patented Molecular Inversion Probe (MIP) technology that combines the specificity of a hybrid capture protocol with the ease of an amplicon-based protocol and utilizes an advanced bioinformatics analysis pipeline based on machine learning. To demonstrate its efficacy, two independent genetic tests were validated on this technology transfer platform: expanded carrier screening (ECS) and hereditary cancer testing (HC). The analytical performance of ECS and HC was validated separately in a blinded manner for calling three different types of variants: SNVs, short indels (typically, <50 bp), and large indels/CNVs defined as multi-exonic del/dup events. The reference set was constructed using samples from Coriell Institute, an external clinical genetic testing laboratory, Maine Molecular Quality Controls Inc. (MMQCI), SeraCare and GIAB Consortium. Overall, the analytical performance showed a sensitivity and specificity of >99.4% for both ECS and HC in detecting SNVs. For indels, both tests reported specificity of 100%, and ECS demonstrated a sensitivity of 100%, whereas HC exhibited a sensitivity of 96.5%. The bioinformatics pipeline also correctly called all reference CNV events resulting in a sensitivity of 100% for both tests. No additional calls were made in the HC panel, leading to a perfect performance (specificity and F-measure of 100%). In the carrier panel, however, three additional positive calls were made outside the reference set. Two of these calls were confirmed using an orthogonal method and were re-classified as true positives leaving only one false positive. The pipeline also correctly identified all challenging carrier statuses, such as positive cases for spinal muscular atrophy and alpha-thalassemia, resulting in 100% sensitivity. After confirmation of additional positive calls via long-range PCR and MLPA, specificity for such cases was estimated at 99%. These performance metrics demonstrate that this tech-transfer solution can be confidently internalized by clinical labs and hospitals to offer mainstream ECS and HC as part of their test catalog, substantially increasing access to quality germline genetic testing for labs of all sizes and resources levels.

Keywords: clinical genetics, genetic testing, molecular genetics, technology transfer

Procedia PDF Downloads 152
532 A Rapid Colorimetric Assay for Direct Detection of Unamplified Hepatitis C Virus RNA Using Gold Nanoparticles

Authors: M. Shemis, O. Maher, G. Casterou, F. Gauffre

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Hepatitis C virus (HCV) is a major cause of chronic liver disease with a global 170 million chronic carriers at risk of developing liver cirrhosis and/or liver cancer. Egypt reports the highest prevalence of HCV worldwide. Currently, two classes of assays are used in the diagnosis and management of HCV infection. Despite the high sensitivity and specificity of the available diagnostic assays, they are time-consuming, labor-intensive, expensive, and require specialized equipment and highly qualified personal. It is therefore important for clinical and economic terms to develop a low-tech assay for the direct detection of HCV RNA with acceptable sensitivity and specificity, short turnaround time, and cost-effectiveness. Such an assay would be critical to control HCV in developing countries with limited resources and high infection rates, such as Egypt. The unique optical and physical properties of gold nanoparticles (AuNPs) have allowed the use of these nanoparticles in developing simple and rapid colorimetric assays for clinical diagnosis offering higher sensitivity and specificity than current detection techniques. The current research aims to develop a detection assay for HCV RNA using gold nanoparticles (AuNPs). Methods: 200 anti-HCV positive samples and 50 anti-HCV negative plasma samples were collected from Egyptian patients. HCV viral load was quantified using m2000rt (Abbott Molecular Inc., Des Plaines, IL). HCV genotypes were determined using multiplex nested RT- PCR. The assay is based on the aggregation of AuNPs in presence of the target RNA. Aggregation of AuNPs causes a color shift from red to blue. AuNPs were synthesized using citrate reduction method. Different sets of probes within the 5’ UTR conserved region of the HCV genome were designed, grafted on AuNPs and optimized for the efficient detection of HCV RNA. Results: The nano-gold assay could colorimetrically detect HCV RNA down to 125 IU/ml with sensitivity and specificity of 91.1% and 93.8% respectively. The turnaround time of the assay is < 30 min. Conclusions: The assay allows sensitive and rapid detection of HCV RNA and represents an inexpensive and simple point-of-care assay for resource-limited settings.

Keywords: HCV, gold nanoparticles, point of care, viral load

Procedia PDF Downloads 178
531 A Ratio-Weighted Decision Tree Algorithm for Imbalance Dataset Classification

Authors: Doyin Afolabi, Phillip Adewole, Oladipupo Sennaike

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Most well-known classifiers, including the decision tree algorithm, can make predictions on balanced datasets efficiently. However, the decision tree algorithm tends to be biased towards imbalanced datasets because of the skewness of the distribution of such datasets. To overcome this problem, this study proposes a weighted decision tree algorithm that aims to remove the bias toward the majority class and prevents the reduction of majority observations in imbalance datasets classification. The proposed weighted decision tree algorithm was tested on three imbalanced datasets- cancer dataset, german credit dataset, and banknote dataset. The specificity, sensitivity, and accuracy metrics were used to evaluate the performance of the proposed decision tree algorithm on the datasets. The evaluation results show that for some of the weights of our proposed decision tree, the specificity, sensitivity, and accuracy metrics gave better results compared to that of the ID3 decision tree and decision tree induced with minority entropy for all three datasets.

Keywords: data mining, decision tree, classification, imbalance dataset

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530 Differences in Activity Patterns between Adult and U-21 Major League Players in Four Field Positions

Authors: U. Harel, E. Carmeli

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The Purpose was to measure differences in activity patterns between major league adult and U-21 soccer players. Four U-21 players and four adult team players were evaluated using a repeated measures technique. All eight players were affiliated with the Maccabi Haifa soccer club from the Israeli professional and U-21major leagues, depending on the player’s age. GPS sensors were attached to the players during five consecutive games to identify patterns regarding running distance and speed according to the field positions. There was no significant difference in the total running distances covered by two age groups. When measuring running speed, an advantage was observed in the adult group when comparing two players from different age groups that played the same position. Differences in activity patterns were evident between adult and U-21 major league soccer players. Furthermore, differences in within group activity pattern emerged between the positions under investigation. These findings provide valuable knowledge that may serve the principle of training specificity.

Keywords: physical fitness, soccer, positional differences, GPS, training specificity

Procedia PDF Downloads 118
529 Non-Invasive Assessment of Peripheral Arterial Disease: Automated Ankle Brachial Index Measurement and Pulse Volume Analysis Compared to Ultrasound Duplex Scan

Authors: Jane E. A. Lewis, Paul Williams, Jane H. Davies

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Introduction: There is, at present, a clear and recognized need to optimize the diagnosis of peripheral arterial disease (PAD), particularly in non-specialist settings such as primary care, and this arises from several key facts. Firstly, PAD is a highly prevalent condition. In 2010, it was estimated that globally, PAD affected more than 202 million people and furthermore, this prevalence is predicted to further escalate. The disease itself, although frequently asymptomatic, can cause considerable patient suffering with symptoms such as lower limb pain, ulceration, and gangrene which, in worse case scenarios, can necessitate limb amputation. A further and perhaps the most eminent consequence of PAD arises from the fact that it is a manifestation of systemic atherosclerosis and therefore is a powerful predictor of coronary heart disease and cerebrovascular disease. Objective: This cross sectional study aimed to individually and cumulatively compare sensitivity and specificity of the (i) ankle brachial index (ABI) and (ii) pulse volume waveform (PVW) recorded by the same automated device, with the presence or absence of peripheral arterial disease (PAD) being verified by an Ultrasound Duplex Scan (UDS). Methods: Patients (n = 205) referred for lower limb arterial assessment underwent an ABI and PVW measurement using volume plethysmography followed by a UDS. Presence of PAD was recorded for ABI if < 0.9 (noted if > 1.30) if PVW was graded as 2, 3 or 4 or a hemodynamically significant stenosis > 50% with UDS. Outcome measure was agreement between measured ABI and interpretation of the PVW for PAD diagnosis, using UDS as the reference standard. Results: Sensitivity of ABI was 80%, specificity 91%, and overall accuracy 88%. Cohen’s kappa revealed good agreement between ABI and UDS (k = 0.7, p < .001). PVW sensitivity 97%, specificity 81%, overall accuracy 84%, with a good level of agreement between PVW and UDS (k = 0.67, p < .001). The combined sensitivity of ABI and PVW was 100%, specificity 76%, and overall accuracy 85% (k = 0.67, p < .001). Conclusions: Combing these two diagnostic modalities within one device provided a highly accurate method of ruling out PAD. Such a device could be utilized within the primary care environment to reduce the number of unnecessary referrals to secondary care with concomitant cost savings, reduced patient inconvenience, and prioritization of urgent PAD cases.

Keywords: ankle brachial index, peripheral arterial disease, pulse volume waveform, ultrasound duplex scan

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528 Structure, Bioinformatics Analysis and Substrate Specificity of a 6-Phospho-β-Glucosidase Glycoside Hydrolase 1 Enzyme from Bacillus licheniformis

Authors: Wayde Veldman, Ozlem T. Bishop, Igor Polikarpov

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In bacteria, mono and disaccharides are phosphorylated during uptake into the cell via the widely used phosphoenolpyruvate (PEP)-dependent phosphotransferase transport system. As an initial step in the phosphorylated disaccharide metabolism pathway, certain glycoside hydrolase family 1 (GH1) enzymes play a crucial role in releasing phosphorylated and non-phosphorylated monosaccharides. However, structural determinants for the specificity of these enzymes still need to be clarified. GH1 enzymes are known to have a wide array of functions. According to the CAZy database, there are twenty-one different enzymatic activities in the GH1 family. Here, the structure and substrate specificity of a GH1 enzyme from Bacillus licheniformis, hereafter known as BlBglH, was investigated. The sequence of the enzyme BlBglH was compared to the sequences of other characterized GH1 enzymes using sequence alignment, sequence identity calculations, phylogenetic analysis, and motif discovery. Through these various analyses, BlBglH was found to have sequence features characteristic of the 6-phospho-β-glucosidase activity enzymes. Additionally, motif and structure comparisons of the three most commonly studied GH1 enzyme-activities revealed a shared loop amongst the different structures that consist of different sequence motifs – this loop is thought to guide specific substrates (depending on activity) towards the active-site. To further affirm BlBglH enzyme activity, molecular docking and molecular dynamics simulations were performed. Docking was carried out using 6-phospho-β-glucosidase enzyme-activity positive (p-Nitrophenyl-beta-D-glucoside-6-phosphate) and negative (p-Nitrophenyl-beta-D-galactoside-6-phosphate) control ligands, followed by 400 ns molecular dynamics simulations. The positive-control ligand maintained favourable interactions within the active site until the end of the simulation. The negative-control ligand was observed exiting the enzyme at 287 ns. Binding free energy calculations showed that the positive-control complex had a substantially more favourable binding energy compared to the negative-control complex. Jointly, the findings of this study suggest that the BlBglH enzyme possesses 6-phospho-β-glucosidase enzymatic activity.

Keywords: 6-P-β-glucosidase, glycoside hydrolase 1, molecular dynamics, sequence analysis, substrate specificity

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527 Experimental Measurements for the Effect of Dilution Procedure in Blood Esterases as Animals Biomarker for Exposure to Organophosphate Compounds

Authors: Kasim Sakran Abass

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This main aim of this study was to confirm and extend our current knowledge about the effects of dilutions on esterases activities in the blood for birds with respect to protecting the enzyme from organophosphate inhibition. There were significantly higher esterases activities in dilution 1:10 in all blood samples from quail, duck, and chick compared to other dilutions (1:5, 1:15, 1:20, and 1:25). Furthermore, our results also pointed to the importance of estimating different dilutions effects prior to using in birds as biomarker tools of environmental exposure. Concentration–inhibition curves were determined for the inhibitor in the presence of dilutions 1:5, 1:10 plus 1:15 (to stimulate carboxylesterase). Point estimates (concentrations calculated to produce 20, 50, and 80% inhibition) were compared across conditions and served as a measure of esterase-mediated detoxification. Among the thiol esters (dilution 1:5) was observed to have the highest specificity constant (kcat/Km), and the Km and kcat values were 176 μM and 16,765 s−1, respectively for S-phenyl thioacetate ester, while detected in (dilution 1:15) the lowest specificity constant (kcat/Km), and the Km and kcat values were 943 μM and 1154 s−1, respectively for acetylthiocholine iodide ester.

Keywords: esterase, animal, dilution, pesticides

Procedia PDF Downloads 497