Measurement of 8-hydroxy-2′-deoxyguanosine in DNA by high-performance liquid chromatography-mass spectrometry: comparison with measurement by gas chromatography-mass spectrometry

Measurement of 8-hydroxy-2′-deoxyguanosine (8-OH-dGuo) in DNA by high-performance liquid chromatography/mass spectrometry (LC/MS) was studied. A methodology was developed for separation by LC of 8-OH-dGuo from intact and modified nucleosides in DNA hydrolyzed by a combination of four enzymes: DNase I, phosphodiesterases I and II and alkaline phosphatase. The atmospheric pressure ionization-electrospray process was used for mass spectral measurements. A stable isotope-labeled analog of 8-OH-dGuo was used as an internal standard for quantification by isotope-dilution MS (IDMS). Results showed that LC/IDMS with selected ion-monitoring (SIM) is well suited for identification and quantification of 8-OH-dGuo in DNA at background levels and in damaged DNA. The sensitivity level of LC/IDMS-SIM was found to be comparable to that reported previously using LC-tandem MS (LC/MS/MS). It was found that approximately five lesions per 106 DNA bases can be detected using amounts of DNA as low as 2 µg. The results also suggest that this lesion may be quantified in DNA at levels of one lesion per 106 DNA bases, or even lower, when more DNA is used. Up to 50 µg of DNA per injection were used without adversely affecting the measurements. Gas chromatography/isotope-dilution MS with selected-ion monitoring (GC/IDMS-SIM) was also used to measure this compound in DNA following its removal from DNA by acidic hydrolysis or by hydrolysis with Escherichia coli Fpg protein. The background levels obtained by LC/IDMS-SIM and GC/IDMS-SIM were almost identical. Calf thymus DNA and DNA isolated from cultured HeLa cells were used for this purpose. This indicates that these two techniques can provide similar results in terms of the measurement of 8-OH-dGuo in DNA. In addition, DNA in buffered aqueous solution was damaged by ionizing radiation at different radiation doses and analyzed by LC/IDMS-SIM and GC/IDMS-SIM. Again, similar results were obtained by the two techniques. The sensitivity of GC/MS-SIM for 7,8-dihydro-8-oxoguanine was also examined and found to be much greater than that of LC/MS-SIM and the reported sensitivity of LC/MS/MS for 8-OH-dGuo. Taken together, the results unequivocally show that LC/IDMS-SIM is well suited for sensitive and accurate measurement of 8-OH-dGuo in DNA and that both LC/IDMS-SIM and GC/IDMS-SIM can provide similar results.

An evaluation of the performance of cDNA microarrays for detecting changes in global mRNA expression

The cDNA microarray is one technological approach that has the potential to accurately measure changes in global mRNA expression levels. We report an assessment of an optimized cDNA microarray platform to generate accurate, precise and reliable data consistent with the objective of using microarrays as an acquisition platform to populate gene expression databases. The study design consisted of two independent evaluations with 70 arrays from two different manufactured lots and used three human tissue sources as samples: placenta, brain and heart. Overall signal response was linear over three orders of magnitude and the sensitivity for any element was estimated to be 2 pg mRNA. The calculated coefficient of variation for differential expression for all non-differentiated elements was 12–14% across the entire signal range and did not vary with array batch or tissue source. The minimum detectable fold change for differential expression was 1.4. Accuracy, in terms of bias (observed minus expected differential expression ratio), was less than 1 part in 10 000 for all non-differentiated elements. The results presented in this report demonstrate the reproducible performance of the cDNA microarray technology platform and the methods provide a useful framework for evaluating other technologies that monitor changes in global mRNA expression.

Hydroperoxide lyase depletion in transgenic potato plants leads to an increase in aphid performance

Hydroperoxide lyases (HPLs) catalyze the cleavage of fatty acid hydroperoxides to aldehydes and oxoacids. These volatile aldehydes play a major role in forming the aroma of many plant fruits and flowers. In addition, they have antimicrobial activity in vitro and thus are thought to be involved in the plant defense response against pest and pathogen attack. An HPL activity present in potato leaves has been characterized and shown to cleave specifically 13-hydroperoxides of both linoleic and linolenic acids to yield hexanal and 3-hexenal, respectively, and 12-oxo-dodecenoic acid. A cDNA encoding this HPL has been isolated and used to monitor gene expression in healthy and mechanically damaged potato plants. HPL gene expression is subject to developmental control, being high in young leaves and attenuated in older ones, and it is induced weakly by wounding. HPL enzymatic activity, nevertheless, remains constant in leaves of different ages and also after wounding, suggesting that posttranscriptional mechanisms may regulate its activity levels. Antisense-mediated HPL depletion in transgenic potato plants has identified this enzyme as a major route of 13-fatty acid hydroperoxide degradation in the leaves. Although these transgenic plants have highly reduced levels of both hexanal and 3-hexenal, they show no phenotypic differences compared with wild-type ones, particularly in regard to the expression of wound-induced genes. However, aphids feeding on the HPL-depleted plants display approximately a two-fold increase in fecundity above those feeding on nontransformed plants, consistent with the hypothesis that HPL-derived products have a negative impact on aphid performance. Thus, HPL-catalyzed production of C6 aldehydes may be a key step of a built-in resistance mechanism of plants against some sucking insect pests.

Increased dopamine turnover in the prefrontal cortex impairs spatial working memory performance in rats and monkeys.

The selective activation of the prefrontal cortical dopamine system by mild stress can be mimicked by anxiogenic beta-carbolines such as FG7142. To investigate the functional relevance of elevated levels of dopamine turnover in the prefrontal cortex, the current study examined the effects of FG7142 on the performance of spatial working memory tasks in the rat and monkey. FG7142 selectively increased prefrontal cortical dopamine turnover in rats and significantly impaired performance on spatial working memory tasks in both rats and monkeys. Spatial discrimination, a task with similar motor and motivational demands (rats), or delayed response performance following zero-second delays (monkeys) was unaffected by FG7142. Further, biochemical analysis in rats revealed a significant positive correlation between dopamine turnover in the prefrontal cortex and cognitive impairment on the delayed alternation task. The cognitive deficits in both rats and monkeys were prevented by pretreatment with the benzodiazepine receptor antagonist, RO15-1788, which blocked the increase in dopamine turnover and by the dopamine receptor antagonists, haloperidol, clozapine, and SCH23390. These findings indicate that excessive dopamine activity in the prefrontal cortex is detrimental to cognitive functions mediated by the prefrontal cortex.

School Principals' Attitudes Towards Performance Based Compensation

The main objective of this study is to determine the attitudes of school principals regarding a performance based compensation system. This study identifies the attitudes towards specific factors that should be considered in the implementation of a system of performance based compensation. The data have been analyzed to determine if a principal's demographic characteristics affect his/her level of agreement with performance based compensation and the factors for implementation. In addition, this study unveils areas of concern that principals have conveyed regarding the implementation of a performance based compensation system. Data was obtained from 444 public school principals representing 444 schools and 178 districts in the state of Colorado. Measures used in the treatment of the data include descriptive statistics and one-way ANOVA. The major findings of this study were: 1. 82.4% of respondents believe that teachers, principals and administrators should be included in performance based compensation (PBC). 2. The top two indicators that respondents believed should be included in a PBC system are student achievement (88.5%) and teacher evaluations (77.6%) 3. The 3 largest obstacles to PBC that respondents identified are: a. The capacity to link student achievement to teacher evaluations (82.9%) b. Teacher Union Resistance (67.1%) c. Cost (55.9%) 4. Principals in urban, rural and suburban geographic groups disagree about the effects of performance based compensation. 5. The top 5 overall concerns regarding Performance Based Compensation were: a. Concerns regarding effectively using assessment to measure performance of all teachers/equity between teachers b. Concerns regarding evaluation (time for principals to learn, consistency from school to school, time for principals to evaluate, quality of evaluation tool). c. Not in favor of PBC due to philosophical views or concerns about lack of research. d. Concerns regarding the equity between classrooms and districts across the state due to poverty levels and unequal resources. e. Concerns that performance based compensation will result in a decline in teacher collaboration and an increase in competition between teachers. Based upon these findings, the researcher concluded that there is not a strong general acceptance of performance based compensation systems. However, urban principals in Colorado tend to view PBC somewhat more favorably than do principals in suburban or rural areas. Most importantly, systems to link student achievement to teacher evaluation must be collaboratively created to ensure PBC systems are equitable, consistent and fair.

A Young Child's Anxiety and Math Performance: Enhancing Cognitive Behavioral Intervention with Conjoint Behavioral Consultation

Partnering with families, school personnel, and community resources is an important step to supporting the child and family, especially when children might suffer from debilitating anxiety concerns. However, little research examines the impact of anxiety on math performance for young children participating in school-based interventions enhanced by family components. The following research questions were addressed in the study: 1a) Will a young child with elevated levels of anxiety show a decrease in anxiety symptoms with a Cognitive Behavioral framework intervention program for children? 1b) Will anxiety be reduced with the addition of a Conjoint Behavioral Consultation with the family and teacher? 2a) Will a young child show an increase in math performance after participation in a Cognitive Behavioral framework intervention program for children? 2b) Will math performance be increased with the addition of a Conjoint Behavioral Consultation with the family and teacher? A single-subject staggered baseline across situations intervention study addressed whether the Coping Cat, an evidenced-based child-focused intervention now widely used in schools and clinics to treat childhood anxiety, combined with family and school consultation will decrease elevated anxiety levels and improve math performance in an elementary-aged student. The objective was to support mental health development and math performance with an eight-year-old, female elementary student through a collaborative effort of stakeholders in the student's life. Baseline data was collected with repeated measures of anxiety and math performance, and was compared to two intervention phases: first, a child-focused intervention and second, a family and school consultation. The study tested the theory that the Cognitive Behavioral intervention and Conjoint Behavioral Consultation intervention will influence, positively, the anxiety levels and math performance for an elementary-aged student. Results indicate that the child participant with elevated levels of anxiety showed a reduction in symptoms with the introduction of a Cognitive Behavioral framework intervention when compared to her baseline data. The participant showed further reduction in symptoms across the school and home settings with the implementation of Conjoint Behavioral Consultation when compared to baseline and the first intervention phase. Math performance began to increase with the introduction of the Cognitive Behavioral intervention, and continued to improve with the implementation of the Conjoint Behavioral Consultation. Findings suggest that consultation should begin immediately when an intervention is implemented in order to enhance outcomes.

Subclinical social anxiety and academic performance in adolescence: analysis of theoretical and practical implications

This study analyses the relationship between self-reported social anxiety and academic performance in a sample of 1,616 Spanish students (52.1% males) in compulsory secondary education, aged 12 to 16 years old. Social anxiety was assessed by the Social Phobia and Anxiety Inventory (SPAI) and academic performance was measured with school grades and failing grades. Results reveal that adolescents with social anxiety show a similar academic performance to adolescents without social anxiety. Although t tests found some significant differences in academic grades and number of failing grades, the effect size analysis showed that these differences had no empirical relevance. These findings are discussed considering the gender and grade levels and their theoretical and practical implication.

Comparative Analysis of the Visual Performance After Cataract Surgery With Implantation of a Bifocal or Trifocal Diffractive IOL

PURPOSE: To evaluate and compare the visual, refractive, contrast sensitivity, and aberrometric outcomes with a diffractive bifocal and trifocal intraocular lens (IOL) of the same material and haptic design. METHODS: Sixty eyes of 30 patients undergoing bilateral cataract surgery were enrolled and randomly assigned to one of two groups: the bifocal group, including 30 eyes implanted with the bifocal diffractive IOL AT LISA 801 (Carl Zeiss Meditec, Jena, Germany), and the trifocal group, including eyes implanted with the trifocal diffractive IOL AT LISA tri 839 MP (Carl Zeiss Meditec). Analysis of visual and refractive outcomes, contrast sensitivity, ocular aberrations (OPD-Scan III; Nidek, Inc., Gagamori, Japan), and defocus curve were performed during a 3-month follow-up period. RESULTS: No statistically significant differences between groups were found in 3-month postoperative uncorrected and corrected distance visual acuity (P > .21). However, uncorrected, corrected, and distance-corrected near and intermediate visual acuities were significantly better in the trifocal group (P < .01). No significant differences between groups were found in postoperative spherical equivalent (P = .22). In the binocular defocus curve, the visual acuity was significantly better for defocus of -0.50 to -1.50 diopters in the trifocal group (P < .04) and -3.50 to -4.00 diopters in the bifocal group (P < .03). No statistically significant differences were found between groups in most of the postoperative corneal, internal, and ocular aberrations (P > .31), and in contrast sensitivity for most frequencies analyzed (P > .15). CONCLUSIONS: Trifocal diffractive IOLs provide significantly better intermediate vision over bifocal IOLs, with equivalent postoperative levels of visual and ocular optical quality.

A three-stage DEA model to evaluate learning-teaching technical efficiency: Key performance indicators and contextual variables

This study evaluates the technical efficiency of the learning-teaching process in higher education using a three-stage procedure that offers advances in comparison to previous studies and improves the quality of the results. First, it utilizes a multiple stage Data Envelopment Analysis (DEA) with contextual variables. Second, the levels of super efficiency are calculated in order to prioritize the efficiency units. And finally, through sensitivity analysis, the contribution of each key performance indicator (KPI) is established with respect to the efficiency levels without omission of variables. The analytical data was collected from a survey completed by 633 tourism students during the 2011/12, 2012/13 and 2013/14 academic course years. The results suggest that level of satisfaction with the course, diversity of materials and satisfaction with the teacher were the most important factors affecting teaching performance. Furthermore, the effect of the contextual variables was found to be significant.

Décès pendant une simulation médicale : perspectives des apprenants et impacts sur le stress et la performance des réanimateurs

L’acceptabilité d’un décès lors d’une simulation médicale reste débattue mais il existe peu de données sur la perspective des apprenants. Des médecins résidents ont effectué une pratique de code et ont rempli un questionnaire pré et post-simulation. Ils ont été exposés à deux scénarios où un bébé naissait sans signe de vie: 1. Nouveau-né ne répondant pas aux manœuvres de réanimation (DCD); 2. Nouveau-né s’améliorant avec une réanimation adéquate (REA). Les performances étaient évaluées à l’aide de la grille standardisée du Programme de Réanimation Néonatale. Le stress objectif (cortisol salivaire) et subjectif a été mesuré après le code. La rétroaction («feedback»), individuelle et en groupe, fut analysée à l’aide de méthodologies qualitatives. 59/62 apprenants ont répondu au questionnaire et 42 ont participé à la simulation. Tous les résidents trouvent les simulations bénéfiques et souhaitent y être exposés davantage. Le type et l’ordre des scénarios n’ont pas eu d’impact sur la performance. Un seul résident a interrompu les manœuvres de réanimation après 10 minutes d’asystolie, tel que recommandé, et 31% ont poursuivi après 20 minutes. Les participants trouvaient le scénario DCD plus stressant. Les niveaux de cortisol salivaire ont augmenté après les simulations (p<0.001) et ce, pour les deux scénarios. Cette augmentation était indépendante du scénario (p=0.06) et n’était pas associée à la performance. Les réponses à la question « Comment a été votre expérience? », ont permis d’identifier deux thèmes: 1. Le mannequin ne meurt pas en simulation médicale; 2. Le décès lors de la simulation signifie une réanimation inadéquate. Le décès lors d’une pratique de code est stressant, mais n’interfère pas avec la performance des soignants. Les apprenants trouvent cet exercice acceptable et bénéfique à leur pratique future.

Influence of experience and training on dental students’ examination performance regarding panoramic images

Physician training has greatly benefitted from insights gained in understanding the manner in which experts search medical images for abnormalities. The aims of this study were to compare the search patterns of 30 fourth-year dental students and 15 certified oral and maxillofacial radiologists (OMRs) over panoramic images and to determine the most robust variables for future studies involving image visualization. Eye tracking was used to capture the eye movement patterns of both subject groups when examining 20 panoramic images classified as normal or abnormal. Abnormal images were further subclassified as having an obvious, intermediate, or subtle abnormality. The images were presented in random order to each participant, and data were collected on duration of the participants’ observations and total distance tracked, time to first eye fixation, and total duration and numbers of fixations on and off the area of interest (AOI). The results showed that the OMRs covered greater distances than the dental students (p<0.001) for normal images. For images of pathosis, the OMRs required less total time (p<0.001), made fewer eye fixations (p<0.01) with fewer saccades (p<0.001) than the students, and required less time before making the first fixation on the AOI (p<0.01). Furthermore, the OMRs covered less distance (p<0.001) than the dental students for obvious pathoses. For investigations of images of pathosis, time to first fixation is a robust parameter in predicting ability. For images with different levels of subtlety of pathoses, the number of fixations, total time spent, and numbers of revisits are important parameters to analyze when comparing observer groups with different levels of experience.

Therapeutic drug monitoring of cefepime with micellar electrokinetic capillary chromatography: Assay improvement, quality assurance, and impact on patient drug levels

The improvement and performance of a micellar electrokinetic capillary chromatography assay for cefepime in human serum and plasma with a 50 μm id fused-silica capillary elongated from 40 to 60 cm is reported. Sample preparation with dodecylsulfate protein precipitation at pH 4.5, the pH 9.1 separation medium and the applied voltage were as reported previously[16]. The change resulted in a significant lower current, higher resolution and increased detection time intervals. The performance of the assay with multi-level internal calibration was assessed with calibration and control samples. Quality assurance data of a two year period assessed under the new conditions demonstrated the robustness of the assay. In serum samples of patients who received both cefepime and sulfamethoxazole, cefepime could not be detected due to the inseparability of the two compounds. The presence of an interference can be recognized by an increased peak width (width > 0.2 min), the appearance of a shoulder or an unresolved double peak. The patient data gathered during a three year period reveal that introduction of therapeutic drug monitoring led to a 50% reduction of the median drug level. The data suggest that therapeutic drug monitoring can help to minimize the risk of major adverse reactions and to increase drug safety on an individual basis. This article is protected by copyright. All rights reserved.

Influence of experience and training on dental students’ examination performance regarding panoramic images.

Physician training has greatly benefitted from insights gained in understanding the manner in which experts search medical images for abnormalities. The aims of this study were to compare the search patterns of 30 fourth-year dental students and 15 certified oral and maxillofacial radiologists (OMRs) over panoramic images and to determine the most robust variables for future studies involving image visualization. Eye tracking was used to capture the eye movement patterns of both subject groups when examining 20 panoramic images classified as normal or abnormal. Abnormal images were further subclassified as having an obvious, intermediate, or subtle abnormality. The images were presented in random order to each participant, and data were collected on duration of the participants’ observations and total distance tracked, time to first eye fixation, and total duration and numbers of fixations on and off the area of interest (AOI). The results showed that the OMRs covered greater distances than the dental students (p<0.001) for normal images. For images of pathosis, the OMRs required less total time (p<0.001), made fewer eye fixations (p<0.01) with fewer saccades (p<0.001) than the students, and required less time before making the first fixation on the AOI (p<0.01). Furthermore, the OMRs covered less distance (p<0.001) than the dental students for obvious pathoses. For investigations of images of pathosis, time to first fixation is a robust parameter in predicting ability. For images with different levels of subtlety of pathoses, the number of fixations, total time spent, and numbers of revisits are important parameters to analyze when comparing observer groups with different levels of experience.

Low-dose alcohol effects on human behavior and performance: a review of post-1984 research.

Federal Aviation Administration, Washington, D.C.

An analysis of ultimate performance measures to determine total project impact in Columbus, Georgia from January 1972 - December 1974. Section II - volume I. Final report.

National Highway Traffic Safety Administration, Office of Driver and Pedestrian Programs, Washington, D.C.