Immunoassays with rolling circle DNA amplification: A versatile platform for ultrasensitive antigen detection

We describe an adaptation of the rolling circle amplification (RCA) reporter system for the detection of protein Ags, termed “immunoRCA.” In immunoRCA, an oligonucleotide primer is covalently attached to an Ab; thus, in the presence of circular DNA, DNA polymerase, and nucleotides, amplification results in a long DNA molecule containing hundreds of copies of the circular DNA sequence that remain attached to the Ab and that can be detected in a variety of ways. Using immunoRCA, analytes were detected at sensitivities exceeding those of conventional enzyme immunoassays in ELISA and microparticle formats. The signal amplification afforded by immunoRCA also enabled immunoassays to be carried out in microspot and microarray formats with exquisite sensitivity. When Ags are present at concentrations down to fM levels, specifically bound Abs can be scored by counting discrete fluorescent signals arising from individual Ag–Ab complexes. Multiplex immunoRCA also was demonstrated by accurately quantifying Ags mixed in different ratios in a two-color, single-molecule-counting assay on a glass slide. ImmunoRCA thus combines high sensitivity and a very wide dynamic range with an unprecedented capability for single molecule detection. This Ag-detection method is of general applicability and is extendable to multiplexed immunoassays that employ a battery of different Abs, each labeled with a unique oligonucleotide primer, that can be discriminated by a color-coded visualization system. ImmunoRCA-profiling based on the simultaneous quantitation of multiple Ags should expand the power of immunoassays by exploiting the increased information content of ratio-based expression analysis.

Mapping hippocampal and ventricular change in Alzheimer disease

We developed an anatomical mapping technique to detect hippocampal and ventricular changes in Alzheimer disease (AD). The resulting maps are sensitive to longitudinal changes in brain structure as the disease progresses. An anatomical surface modeling approach was combined with surface-based statistics to visualize the region and rate of atrophy in serial MRI scans and isolate where these changes link with cognitive decline. Fifty-two high-resolution MRI scans were acquired from 12 AD patients (age: 68.4 +/- 1.9 years) and 14 matched controls (age: 71.4 +/- 0.9 years), each scanned twice (2.1 +/- 0.4 years apart). 3D parametric mesh models of the hippocampus and temporal horns were created in sequential scans and averaged across subjects to identify systematic patterns of atrophy. As an index of radial atrophy, 3D distance fields were generated relating each anatomical surface point to a medial curve threading down the medial axis of each structure. Hippocampal atrophic rates and ventricular expansion were assessed statistically using surface-based permutation testing and were faster in AD than in controls. Using color-coded maps and video sequences, these changes were visualized as they progressed anatomically over time. Additional maps localized regions where atrophic changes linked with cognitive decline. Temporal horn expansion maps were more sensitive to AD progression than maps of hippocampal atrophy, but both maps correlated with clinical deterioration. These quantitative, dynamic visualizations of hippocampal atrophy and ventricular expansion rates in aging and AD may provide a promising measure to track AD progression in drug trials. (C) 2004 Elsevier Inc. All rights reserved.

Three-dimensional modeling of the human eye based on magnetic resonance imaging

PURPOSE. A methodology for noninvasively characterizing the three-dimensional (3-D) shape of the complete human eye is not currently available for research into ocular diseases that have a structural substrate, such as myopia. A novel application of a magnetic resonance imaging (MRI) acquisition and analysis technique is presented that, for the first time, allows the 3-D shape of the eye to be investigated fully. METHODS. The technique involves the acquisition of a T2-weighted MRI, which is optimized to reveal the fluid-filled chambers of the eye. Automatic segmentation and meshing algorithms generate a 3-D surface model, which can be shaded with morphologic parameters such as distance from the posterior corneal pole and deviation from sphericity. Full details of the method are illustrated with data from 14 eyes of seven individuals. The spatial accuracy of the calculated models is demonstrated by comparing the MRI-derived axial lengths with values measured in the same eyes using interferometry. RESULTS. The color-coded eye models showed substantial variation in the absolute size of the 14 eyes. Variations in the sphericity of the eyes were also evident, with some appearing approximately spherical whereas others were clearly oblate and one was slightly prolate. Nasal-temporal asymmetries were noted in some subjects. CONCLUSIONS. The MRI acquisition and analysis technique allows a novel way of examining 3-D ocular shape. The ability to stratify and analyze eye shape, ocular volume, and sphericity will further extend the understanding of which specific biometric parameters predispose emmetropic children subsequently to develop myopia. Copyright © Association for Research in Vision and Ophthalmology.

Heteronormativity and teaching: A phenomenological study of lesbian teachers

The purpose of this phenomenological study was to understand what impact “heteronormativity” has on a lesbian teacher's perception of her instructional style, content, and context of curriculum taught. Through taped interviews with lesbian educators, this research examined the lived experience of the lesbian teacher. The framework for this study included theories related to historical, sociocultural, and psychosocial development while the methodology included a qualitative design using primary elements of a phenomenological study outlined without ignoring the influence associated with contextualism. Due to the sensitive nature of the study nine women who were the focus of this research were volunteers with the first serving as a “gatekeeper” to assist in the pilot study. The subsequent group evolved as a result of “snowballing” to gain more participants. ^ The data in the form of narrative derived from the interviews was transcribed, color-coded, and organized into four themes and associated sub-themes, based upon the perceptions of these educators. These themes characterized the coming out process of a lesbian, which directly paralleled the personal and professional development of the lesbian educator, emerged as a result of the analysis. They included: (a) self-acknowledgement; (b) self-indentification; (c) coming out to other lesbians by overcoming fear and establishing relationships; (d) coming out to others by overcoming heteronormativity by using support groups in defining a lesbian's role as a teacher. ^ The results of this study showed that the acceptance of the lesbian culture, shared with the acknowledgement, rather than compliance or defiance, of cultural hegemony can allow the lesbian educator to develop a curriculum and a classroom climate that will foster understanding and even generate social change among colleagues, parents, and students, one person at a time. ^

An analysis of teachers' perceptions of informal science professional development

Science professional development, which is fundamental to science education improvement, has been described as being weak and fragmentary. The purpose of this study was to investigate teachers' perceptions of informal science professional development to gain an in-depth understanding of the essence of the phenomenon and related science-teaching dispositions. Based on the frameworks of phenomenology, constructivism, and adult learning theory, the focus was on understanding how the phenomenon was experienced within the context of teachers' everyday world. ^ Data were collected from eight middle-school teachers purposefully selected because they had participated in informal programs during Project TRIPS (Teaching Revitalized Through Informal Programs in Science), a collaboration between the Miami-Dade school district, government agencies (including NASA), and non-profit organizations (including Audubon of Florida). In addition, the teachers experienced hands-on labs offered through universities (including the University of Arizona), field sites, and other agencies. ^ The study employed Seidman's (1991) three-interview series to collect the data. Several methods were used to enhance the credibility of the research, including using triangulation of the data. The interviews were transcribed, color-coded and organized into six themes that emerged from the data. The themes included: (a) internalized content knowledge, (b) correlated hands-on activities, (c) enhanced science-teaching disposition, (d) networking/camaraderie, (e) change of context, and (f) acknowledgment as professionals. The teachers identified supportive elements and constraints related to each theme. ^ The results indicated that informal programs offering experiential learning opportunities strengthened understanding of content knowledge. Teachers implemented hands-on activities that were explicitly correlated to their curriculum. Programs that were conducted in a relaxed context enhanced teachers' science-teaching dispositions. However, a lack of financial and administrative support, perceived safety risks, insufficient reflection time, and unclear itineraries impeded program implementation. The results illustrated how informal educators can use this cohesive model as they develop programs that address the supports and constraints to teachers' science instruction needs. This, in turn, can aid teachers as they strive to provide effective science instruction to students; notions embedded in reforms. Ultimately, this can affect how learners develop the ability to make informed science decisions that impact the quality of life on a global scale. ^

Test of a smock system on CPR primary emergency measures and medical errors during simulated emergencies

Rates of survival of victims of sudden cardiac arrest (SCA) using cardio pulmonary resuscitation (CPR) have shown little improvement over the past three decades. Since registered nurses (RNs) comprise the largest group of healthcare providers in U.S. hospitals, it is essential that they are competent in performing the four primary measures (compression, ventilation, medication administration, and defibrillation) of CPR in order to improve survival rates of SCA patients. The purpose of this experimental study was to test a color-coded SMOCK system on: 1) time to implement emergency patient care measures 2) technical skills performance 3) number of medical errors, and 4) team performance during simulated CPR exercises. The study sample was 260 RNs (M 40 years, SD=11.6) with work experience as an RN (M 7.25 years, SD=9.42).Nurses were allocated to a control or intervention arm consisting of 20 groups of 5-8 RNs per arm for a total of 130 RNs in each arm. Nurses in each study arm were given clinical scenarios requiring emergency CPR. Nurses in the intervention group wore different color labeled aprons (smocks) indicating their role assignment (medications, ventilation, compression, defibrillation, etc) on the code team during CPR. Findings indicated that the intervention using color-labeled smocks for pre-assigned roles had a significant effect on the time nurses started compressions (t=3.03, p=0.005), ventilations (t=2.86, p=0.004) and defibrillations (t=2.00, p=.05) when compared to the controls using the standard of care. In performing technical skills, nurses in the intervention groups performed compressions and ventilations significantly better than those in the control groups. The control groups made significantly (t=-2.61, p=0.013) more total errors (7.55 SD 1.54) than the intervention group (5.60, SD 1.90). There were no significant differences in team performance measures between the groups. Study findings indicate use of colored labeled smocks during CPR emergencies resulted in: shorter times to start emergency CPR; reduced errors; more technical skills completed successfully; and no differences in team performance.

Processing the Intracardiac Electrogram for Atrial Fibrillation Ablation

Atrial fibrillation (AF) is a major global health issue as it is the most prevalent sustained supraventricular arrhythmia. Catheter-based ablation of some parts of the atria is considered an effective treatment of AF. The main objective of this research is to analyze atrial intracardiac electrograms (IEGMs) and extract insightful information for the ablation therapy. Throughout this thesis we propose several computationally efficient algorithms that take streams of IEGMs from different atrial sites as the input signals, sequentially analyze them in various domains (e.g., time and frequency), and create color-coded three-dimensional map of the atria to be used in the ablation therapy.

Test of a Smock System on CPR Primary Emergency Measures and Medical Errors During Simulated Emergencies

Rates of survival of victims of sudden cardiac arrest (SCA) using cardio pulmonary resuscitation (CPR) have shown little improvement over the past three decades. Since registered nurses (RNs) comprise the largest group of healthcare providers in U.S. hospitals, it is essential that they are competent in performing the four primary measures (compression, ventilation, medication administration, and defibrillation) of CPR in order to improve survival rates of SCA patients. The purpose of this experimental study was to test a color-coded SMOCK system on:1) time to implement emergency patient care measures 2) technical skills performance 3) number of medical errors, and 4) team performance during simulated CPR exercises. The study sample was 260 RNs (M 40 years, SD=11.6) with work experience as an RN (M 7.25 years, SD=9.42).Nurses were allocated to a control or intervention arm consisting of 20 groups of 5-8 RNs per arm for a total of 130 RNs in each arm. Nurses in each study arm were given clinical scenarios requiring emergency CPR. Nurses in the intervention group wore different color labeled aprons (smocks) indicating their role assignment (medications, ventilation, compression, defibrillation, etc) on the code team during CPR. Findings indicated that the intervention using color-labeled smocks for pre-assigned roles had a significant effect on the time nurses started compressions (t=3.03, p=0.005), ventilations (t=2.86, p=0.004) and defibrillations (t=2.00, p=.05) when compared to the controls using the standard of care. In performing technical skills, nurses in the intervention groups performed compressions and ventilations significantly better than those in the control groups. The control groups made significantly (t=-2.61, p=0.013) more total errors (7.55 SD 1.54) than the intervention group (5.60, SD 1.90). There were no significant differences in team performance measures between the groups. Study findings indicate use of colored labeled smocks during CPR emergencies resulted in: shorter times to start emergency CPR; reduced errors; more technical skills completed successfully; and no differences in team performance.

Extrinsic compression of popliteal artery in asymptomatic athlete and non-athlete individuals - A comparative study using duplex scan (color duplex sonography)

Aim. Extrinsic compression of the popliteal artery and absence of surrounding anatomical abnormalities characterize the functional popliteal artery entrapment syndrome (PAES). The diagnosis is confirmed to individuals who have typical symptoms of popliteal entrapment and occlusion or important stenosis of the popliteal artery with color duplex sonography (CDS), magnetic resonance imaging (MRI) or arteriography during active plantar flexion-extension maneuvers. However, variable result findings in normal asymptomatic subjects have raised doubts as to the validity of these tests. The purpose of this study was to compare the frequency of popliteal artery compression in 2 groups of asymptomatic subjects, athletes and non-athletes.Methods. Forty-two individuals were studied. Twenty-one subjects were indoor soccer players, and 21 were sedentary individuals. Physical activity was evaluated through questionnaires, anthropometric measurements, and cardiopulmonary exercise test. Evaluation of popliteal artery compression was performed in lower limbs with CDS, ankle-brachial index (ABI) measurements and continuous wave Doppler of the posterior tibial artery.Results. The athletes studied fulfilled the criteria of high level of physical activity whereas sedentary subjects met the criteria of low level of activity. Popliteal artery compression was observed with CDS in 6 (14.2%) studied subjects; 2 of whom (4.7%) were athletes and 4 (9.5%) were non-athletes. This difference was not statistically significant (p=0.21). Doppler of the tibial arteries and ABI measurements gave good specificity and sensibility in the identification of popliteal artery compression.Conclusion. The frequency of popliteal artery compression during maneuvers in normal subjects was 14.2% irrespective of whether or not they performed regular physical activities. Both Doppler and ABI showed good agreement with CDS and should be considered in screening popliteal arteries in individuals suspected of PAES.

Novel gray coded pattern for unwrapping phase in fringe projection based 3D profiling

A method to reliably extract object profiles even with height discontinuities (that leads to 2n pi phase jumps) is proposed. This method uses Fourier transform profilometry to extract wrapped phase, and an additional image formed by illuminating the object of interest by a novel gray coded pattern for phase unwrapping. Simulation results suggest that the proposed approach not only retains the advantages of the original method, but also contributes significantly in the enhancement of its performance. Fundamental advantage of this method stems from the fact that both extraction of wrapped phase and unwrapping the same were done by gray scale images. Hence, unlike the methods that use colors, proposed method doesn't demand a color CCD camera and is ideal for profiling objects with multiple colors.

Trellis coded modulation scheme for the fading relay channel

A decode and forward protocol based Trellis Coded Modulation (TCM) scheme for the half-duplex relay channel, in a Rayleigh fading environment, is presented. The proposed scheme can achieve any spectral efficiency greater than or equal to one bit per channel use (bpcu). A near-ML decoder for the suggested TCM scheme is proposed. It is shown that the high Signal to Noise Ratio (SNR) performance of this near-ML decoder approaches the performance of the optimal ML decoder. Based on the derived Pair-wise Error Probability (PEP) bounds, design criteria to maximize the diversity and coding gains are obtained. Simulation results show a large gain in SNR for the proposed TCM scheme over uncoded communication as well as the direct transmission without the relay.

Implementation of a robust coded structured light technique for dynamic 3D measurements

This paper presents the implementation details of a coded structured light system for rapid shape acquisition of unknown surfaces. Such techniques are based on the projection of patterns onto a measuring surface and grabbing images of every projection with a camera. Analyzing the pattern deformations that appear in the images, 3D information of the surface can be calculated. The implemented technique projects a unique pattern so that it can be used to measure moving surfaces. The structure of the pattern is a grid where the color of the slits are selected using a De Bruijn sequence. Moreover, since both axis of the pattern are coded, the cross points of the grid have two codewords (which permits to reconstruct them very precisely), while pixels belonging to horizontal and vertical slits have also a codeword. Different sets of colors are used for horizontal and vertical slits, so the resulting pattern is invariant to rotation. Therefore, the alignment constraint between camera and projector considered by a lot of authors is not necessary

Influência da potência de laser Nd: YAG na soldagem do aço inoxidável duplex UNS S32205

Pós-graduação em Engenharia Mecânica - FEIS

Sunglasses, traffic signals, and color vision deficiencies

Purpose: To determine (a) the effect of different sunglass tint colorations on traffic signal detection and recognition for color normal and color deficient observers, and (b) the adequacy of coloration requirements in current sunglass standards. Methods: Twenty color-normals and 49 color-deficient males performed a tracking task while wearing sunglasses of different colorations (clear, gray, green, yellow-green, yellow-brown, red-brown). At random intervals, simulated traffic light signals were presented against a white background at 5° to the right or left and observers were instructed to identify signal color (red/yellow/green) by pressing a response button as quickly as possible; response times and response errors were recorded. Results: Signal color and sunglass tint had significant effects on response times and error rates (p < 0.05), with significant between-color group differences and interaction effects. Response times for color deficient people were considerably slower than color normals for both red and yellow signals for all sunglass tints, but for green signals they were only noticeably slower with the green and yellow-green lenses. For most of the color deficient groups, there were recognition errors for yellow signals combined with the yellow-green and green tints. In addition, deuteranopes had problems for red signals combined with red-brown and yellow-brown tints, and protanopes had problems for green signals combined with the green tint and for red signals combined with the red-brown tint. Conclusions: Many sunglass tints currently permitted for drivers and riders cause a measurable decrement in the ability of color deficient observers to detect and recognize traffic signals. In general, combinations of signals and sunglasses of similar colors are of particular concern. This is prima facie evidence of a risk in the use of these tints for driving and cautions against the relaxation of coloration limits in sunglasses beyond those represented in the study.