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Sensitive detection of potato spindle tuber and temperate fruit tree viroids by reverse transcription-polymerase chain reaction-probe capture hybridization.

A rapid and sensitive assay for the specific detection of plant viroids using reverse transcription-polymerase chain reaction (RT-PCR) -probe capture hybridization (RT-PCR-enzyme-linked immunosorbent assay (ELISA)) was developed. The assay was applied successfully for the detection of potato spindle tuber viroid, peach latent mosaic viroid, or apple scar skin viroid from viroid infected leaf tissue. Clarified sap extract from infected leaf tissue was treated first with GeneReleaser polymeric matrix to remove inhibitors of RT-PCR reactions. Viroid cDNA was then synthesized and amplified using viroid specific primers in RT-PCR assays and the amplified viroid cDNA (amplicon) was digoxigenin (DIG) -labelled during the amplification process. The amplicon was then detected in a colorimetric hybridization system in a microtiter plate using a biotinylated cDNA capture probe. This system combines the specificity of molecular hybridization, the ease of the colorimetric protocol, and is at least 100-fold more sensitive than gel electrophoretic analysis in detecting the amplified product. Viroid cRNA may replace viroid cDNA as the capture probe. The cRNA probe was several fold more sensitive than the cDNA probe for viroid detection. Six to seven hours are needed to complete the RT-PCR-ELISA for viroid detection from infected leaf tissue.

DNA Probes↗

Magnetic capture-hybridization method for purification and probing of mRNA for neutral protease of Bacillus cereus.

A magnetic capture-hybridization method was assessed for the isolation of prokaryotic mRNA for the neutral protease of B. cereus from liquid culture. A biotin-labeled specific probe was hybridized to the mRNA transcripts and subsequently captured by streptavidin-coated paramagnetic beads. mRNA was detected by dot-blot hybridization with a ds DIG-labeled DNA-probe. The magnetic capture hybridization is a rapid and simple method and has a promising potential for gene expression studies in complex samples.

Bacillus cereus↗

Antiserum generated by DNA vaccine binds to hepatitis E virus (HEV) as determined by PCR and immune electron microscopy (IEM): application for HEV detection by affinity-capture RT-PCR.

Previously, we have described that injection of an expression vector containing hepatitis E virus (HEV) open reading frame 2 (HEV-ORF-2) generated a strong antibody response in mice. To characterize the reaction of this antiserum with native HEV and to evaluate its potential diagnostic application, we tested the antiserum's ability to bind HEV using immune electron microscope (IEM) and affinity-capture reverse transcription polymerase chain reaction (RT-PCR) amplification. Antiserum to ORF-2 aggregated HEV virions as seen by electron microscopy, providing direct evidence that ORF-2 encodes a structural protein. Antiserum also captured HEV for RT-PCR amplification. This antiserum bound HEV from diverse origins (Asia, Africa, Mexico) at virus concentrations found in patient fecal specimens and bile from inoculated non-human primates. The specificity of the affinity binding was demonstrated when pre-immune sera or sera collected from mice injected with control DNA vector (lacking the HEV ORF-2 gene) failed to bind HEV for RT-PCR amplification and IEM. Specific RT-PCR amplification was confirmed by restriction enzyme digestion of PCR products. The sensitivity of the binding was evaluated by RT-PCR amplification of serially diluted bile containing a genetically divergent HEV, Mexico'86. HEV was detected in a 10(-8) dilution of this bile. This is the first report that antibodies elicited by a DNA vaccine recognize native HEV. Our results indicate that ORF-2 encodes a structural protein and that antiserum to this protein enables simple, sensitive, and specific HEV detection by affinity-capture RT-PCR.

Animals↗

[Estimation with the capture-recapture method of the number of economic immigrants in Mallorca].

AIM: estimate the number of irregular economic immigrants in Mallorca. DESIGN: We used the capture-recapture method, an indirect method based on contrasts of data from two or more sources. Data were obtained from the Delegación de Gobierno (police and immigration authority), Comisiones Obreras (labor union), and institutions that provide health-related services to immigrants. Individuals were identified by birth date and country of origin. RESULTS: The total number of economic immigrants estimated with this method was 39 392. According to the Delegación de Gobierno data, the number of regular immigrants on the date of our inquiry was 9000. With the capture-recapture method, the number of irregular immigrants in Mallorca was therefore estimated at 30 000. CONCLUSIONS: The capture-recapture method can be useful to estimate the population of irregular immigrants in a given area at a given time, if sufficiently precise information on the identity of each individual can be obtained.

Delivery of Health Care↗

14-3-3 in the cerebrospinal fluid of patients with variant and sporadic Creutzfeldt-Jakob disease measured using capture assay able to detect low levels of 14-3-3 protein.

A protein capture assay was used to measure 14-3-3 (-isoform) in the cerebrospinal fluid (CSF) of patients with either variant or sporadic Creutzfeldt-Jakob disease (CJD). The results were compared with those obtained using Western blotting. Elevated levels of 14-3-3 were found in 58% of variant CJD (vCJD) patients and 82% of sporadic CJD (spCJD) patients using the protein capture assay. Using a Western blotting technique, the presence of CSF 14-3-3 was detected in 58% of vCJD patients and in 89% of spCJD patients. When the results from the protein capture assay and the Western blot were combined, 14-3-3 was detected in 77% of vCJD patients and in 91% of spCJD patients. These results suggest that although analysis of CSF 14-3-3 is not as useful in vCJD as it is in the sporadic form of the disease, a combination of these two techniques results in increased sensitivity of 14-3-3 for the diagnosis of vCJD.

14-3-3 Proteins↗

Optic capture of the AcrySof intraocular lens in pediatric cataract surgery.

PURPOSE: To assess the results of acrylic intraocular lens (IOL) optic capture in children with cataract. SETTING: Department of Ophthalmology, Hospital de Clínicas José de San Martín, and Instituto de la Vision, School of Medicine, University of Buenos Aires, Argentina. METHODS: Eight children had cataract surgery. After lens and cortex aspiration, an AcrySof (Alcon) IOL was implanted in the bag. A primary posterior capsulorhexis was performed. The optic edges were slipped through the posterior capsule leaflets. Clarity of the visual axis, preoperative and postoperative best corrected visual acuities (BCVAs), and refraction were evaluated. RESULTS: The visual axis remained clear in all cases. No case required a secondary procedure. The mean preoperative BCVA was 0.06 +/- 0.06 (SD). Postoperatively, the mean BCVA was 0.88 +/- 0.11 and the mean spherical equivalent, +0.62 +/- 1.31. The mean follow-up was 28.9 +/- 5.3 months. CONCLUSION: Results show that the optic of an acrylic IOL may be captured through a posterior capsulorhexis in pediatric cataract surgery, combining the advantages of optic capture with a smaller incision and a decreased inflammatory response.

Acrylic Resins↗

Necessity of vitrectomy when optic capture is performed in children older than 5 years.

PURPOSE: To determine whether anterior vitrectomy is necessary when optic capture is performed in children between 5 and 12 years old with congenital cataract. SETTING: Iladevi Cataract and IOL Research Center, Ahmedabad, India. METHODS: This prospective randomized controlled study comprised 41 eyes of 25 children whose mean age was 83.57 months (range 60 to 144 months). Intraocular lens (IOL) implantation with optic capture through a primary posterior continuous curvilinear capsulorhexis was performed in all the eyes. The IOL haptics were bag fixated. Patients were randomly assigned to 1 of 2 groups. Vitrectomy was performed in 1 group (n = 21 eyes) and not performed in the other group (n = 20 eyes). The mean follow-up was 21.04 months. A Student t test and chi-square test were used for statistical analysis. RESULTS: All eyes in the vitrectomy group and 30% in the no-vitrectomy group had a clear visual axis at the last follow-up (P <.001) The visual axis was obscured as a result of anterior vitreous fibrosis in 70% of eyes in the no-vitrectomy group. High-contrast visual acuity was not significantly different between groups (P =.28). Low-contrast sensitivity was significantly better in the vitrectomy group (P =.02). Eighteen eyes (85.7%) in the vitrectomy group and 16 eyes (80%) in the no-vitrectomy group developed deposits on the IOL (P =.62). The deposits were present at the last follow-up in 4 eyes (19.0%) in the vitrectomy group and in 6 eyes (30.0%) in the no-vitrectomy group (P =.85). Three eyes (14.3%) in the vitrectomy group and 8 eyes (40.0%) in the no-vitrectomy group developed synechias (P =.06). CONCLUSION: The results suggest that anterior vitrectomy is necessary with optic capture in children with congenital cataract who are between 5 and 12 years old.

Capsulorhexis↗

Posterior continuous curvilinear capsulorhexis and optic capture of the intraocular lens to prevent secondary opacification in pediatric cataract surgery.

PURPOSE: To evaluate the safety and efficacy of posterior continuous curvilinear capsulorhexis with optic capture n preventing secondary opacification of the visual axis in pediatric eyes having cataract surgery and intraocular lens (IOL) implantation. SETTING: Gimbel Eye Centre, Calgary, Alberta, Canada. METHOD: Posterior continuous curvilinear capsulorhexis with optic capture of the IOL was performed in 18 of 19 consecutive pediatric cataract patients (ages 2 1/2 to 12 years). Heparin-coated IOLs with 6 degree angulation of the haptics were implanted in all eyes. Only eyes with a minimum of 18 months follow-up (n = 16) were analyzed. RESULTS: Mean follow-up in the 16 eyes was 35.5 months +/- 9.45 (SD), ranging from 19 to 49 months. To date, the visual axis has remained clear in all eyes. No anterior vitrectomy was purposefully performed in any eye. CONCLUSION: Posterior continuous curvilinear capsulorhexis with optic capture of the heparin-coated IOL appeared to successfully prevent secondary opacification of the visual axis in pediatric cataract cases.

Cataract↗

Improvements to the photon-production data for radiative capture in ENDF

High-quality photon-production data from thermal-neutron-capture reactions are important for many applications, including oil-well logging, planetary gamma-ray spectroscopy, and environmental techniques. Radiation transport codes usually access photon-production data from the national U.S. nuclear database, ENDF/B. To improve the photon-production data for thermal-neutron capture, we have compiled and evaluated the energies and yields of gamma rays for most naturally occurring isotopes with Z < 31 and for a few heavier isotopes. The impact of improved photon-production cross sections for thermal-neutron capture in Cl is illustrated by comparing MCNP simulations and experimental measurements for a pulsed neutron oil-well logging instrument. The new compilation of photon-production data is being incorporated into the latest ENDF/B evaluations for each isotope and will be provided for future releases of the ENDF/B database.

Journal Article↗

Standardization of electron-capture radionuclides by liquid scintillation counting

Liquid scintillation counting has proven to be an excellent technique to standardize beta-ray emitters with good accuracy. However, the success achieved in beta-ray nuclide standardization is not extensible for radionuclides decaying by electron-capture. Last comparison of 55Fe revealed discrepancies of 5% among laboratories. This paper presents two new versions of the computer program EMI. First we consider a better simulation of the photoelectric interaction of X-rays with the scintillator cocktail (EMI 1.2); and second, a more sophisticated KL1L2L3M atomic rearrangement model (EMI 2.0). We emphasize the importance of improving the simulation of the electron capture process to achieve better standardization of electron-capture nuclides by the CIEMAT/NIST method.

Journal Article↗

Direct analysis of laser capture microdissected cells by MALDI mass spectrometry.

Laser capture microdissection (LCM) has become an important tool in biological research, permitting isolation of specific cell populations from frozen tissue samples containing a mixture of cell types. Cells obtained by LCM can be directly analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). We report here methodology for the preparation and analysis of LCM captured cells with MALDI MS, giving high sensitivity and mass resolution. Comparison of the spectra obtained from cell populations of interest can identify unique disease or function-related protein markers. Using this approach, mass spectra obtained from human breast tissue containing invasive mammary carcinoma and normal breast epithelium using LCM were compared. Over 40 peaks were identified that significantly differed in intensity between invasive mammary carcinoma and normal breast epithelium. In addition, mass spectra are presented that show protein patterns from mouse liver and mouse colon crypts. The reported tissue preparation procedure and subsequent analysis by MALDI MS provide a new methodology for protein discovery involving LCM captured cells.

Animals↗

Application of capture-recapture methodology to estimate the prevalence of dementia in South Carolina.

PURPOSE: The purpose of this study was to estimate the prevalence of dementia in individuals 65 years of age and older in the state of South Carolina using capture-recapture methodology. METHODS: We linked data from the Department of Mental Health admissions, inpatient admissions, and emergency room visits. Separate log-linear models were used to obtain estimates of the underascertainment-corrected prevalence of dementia in twelve age-gender-race subgroups, which were summed to estimate the prevalence of dementia in the total population. RESULTS: We found an overall prevalence of dementia of 14% in South Carolina for persons 65 years of age and older using capture-recapture methodology. This estimate of persons with dementia is 25% higher than the identified cases of dementia in the South Carolina Alzheimer's Disease Registry (10.5%). CONCLUSIONS: Although capture-recapture methods are prone to limitations, they can be used to more accurately estimate the prevalence of dementia in a geographic area.

Aged↗

Clover leaf inferior vena cava filter: in vitro evaluation of filter deployment and comparison of emboli-capturing ability.

A new self-centering stainless steel inferior vena cava filter (clover leaf filter) that can be delivered percutaneously through a 10-F catheter has been developed. The filter is loaded into an angiographic catheter as a set of straightened wires that are mechanically deformed into a predetermined clover leaf shape when the device is delivered. The filter can be deployed easily and effectively into the simulated vena cava. Emboli-capturing efficiency of the new filter was compared in vitro to that of the Greenfield and Bird's Nest inferior vena cava filters. All three filters captured all large 5 x 100-mm, potentially fatal emboli. The clover leaf filter captures more of the smaller (5 x 20-mm and 3 x 20-mm) emboli than the Greenfield filter and less than the Bird's Nest filter.

Embolism↗

What we see: inattention and the capture of attention by meaning.

Attention is necessary for the conscious perception of any object. Objects not attended to are not seen. What is it that captures attention when we are engaged in some attention-absorbing task? Earlier research has shown that there are only a very few stimuli which have this power and therefore are reliably detected under these conditions (for example, Mack & Rock, 1998; Moray, 1959). The two most reliable are the observer's own name and a happy face icon which seem to capture attention by virtue of their meaning. Three experiments are described which explore whether these stimuli are detected under conditions, heretofore unexamined, which either cause inattentional blindness or are associated with a perceptual failure associated with the limits of attention. The evidence obtained indicates that these stimuli have a unique capacity to capture and extend the limits of attention under conditions in which this has been deemed highly unlikely.

Adult↗

Analysis of dextrorphan, a metabolite of dextromethorphan, using gas chromatography with electron-capture detection.

Dextromethorphan, a constituent of many over-the-counter cough syrups, is used as a probe drug for phenotyping subjects for their cytochrome P450 2D6 (CYP2D6) enzyme activity and for measuring CYP2D6 activity of preparations such as microsomes. In such studies, formation of the metabolite dextrorphan is used as indicator of the activity of this CYP enzyme. The present report describes an electron-capture gas chromatographic procedure developed for detection and quantification of dextrorphan in human liver microsomal preparations in vitro. After basification of the incubation mixture, dextrorphan was derivatized with pentafluorobenzoyl chloride under aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column, an electron capture detector, and a printer-integrator. Para-hydroxymephenytoin was carried through the procedure as internal standard. The procedure, which involves the derivatization of dextrorphan under aqueous conditions, is rapid and involves the use of the relatively economical procedure of electron-capture gas chromatography. The derivative is stable and possesses excellent chromatographic properties.

Benzoates↗

Using structured text and templates to capture health status outcomes in the electronic health record.

BACKGROUND: Because of the difficulty of implementing clinical outcomes-focused quality improvement (QI) projects, most organizations' efforts and comparative quality report cards have centered on structure and process indicators or on outcomes captured in administrative data systems. Ultimately, however, health care is intended to improve health. Health status is now frequently included in the set of quality of care information requested by purchasers of health care, accrediting and governmental agencies, and consumer groups. Subsequently, users of electronic health record (EHR) systems are demanding that the EHR support the collection and comparative analysis of health status information over time and by clinical population and provider. APPROACH: As opposed to the current practice in most organizations of collecting health status data via an annual mail survey, the approach discussed in this article utilizes standardized coding and classification (SCC) systems and standardized surveys to capture health status information in the EHR during the routine course of care delivery through the use of templates built on structured text. This method is illustrated with examples from the WAVE EHR. The EHR-based structured text and template approach facilitates the incorporation of health status measurement into the documentation of the patient-provider encounter, builds on recognized SCC systems and standardized surveys, and enables information retrieval for a variety of analyses, including those focused on QI. CONCLUSIONS: Health status outcomes are an essential component of an information set focused on health care quality. To routinely capture and analyze health status variables, SCC systems and standardized health status surveys are necessary.

Forms and Records Control↗

Using quality function deployment to capture the voice of the customer and translate it into the voice of the provider.

BACKGROUND: Health care has a number of historical barriers to capturing the voice of the customer and to incorporating customer wants into health care services, whether the customer is a patient, an insurer, or a community. Quality function deployment (QFD) is a set of tools and practices that can help overcome these barriers to form a process for the planning and design or redesign of products and services. The goal of the project was to increase referral volume and to improve a rehabilitation hospital's capacity to provide comprehensive medical and/or legal evaluations for people with complex and catastrophic injuries or illnesses. HIGH-LEVEL VIEW OF QFD AS A PROCESS: The steps in QFD are as follows: capture of the voice of the customer, quality deployment, functions deployment, failure mode deployment, new process deployment, and task deployment. The output of each step becomes the input to a matrix tool or table of the next step of the process. CASE STUDY EXAMPLE: In 3 1/2 months a nine-person project team at Continental Rehabilitation Hospital (San Diego) used QFD tools to capture the voice of the customer, use these data as the basis for a questionnaire on important qualities of service from the customer's perspective, obtain competitive data on how the organization was perceived to be meeting the demanded qualities, identify measurable dimensions and targets of these qualities, and incorporate the functions and tasks into the delivery of service which are necessary to meet the demanded qualities. DISCUSSION: The future of providing health care services will belong to organizations that can adapt to a rapidly changing environment and to demands for new products and services that are produced and delivered in new ways.

Arizona↗

Immobilized peptides as high-affinity capture agents for self-associating proteins.

There is currently great interest in the fabrication of protein-detecting arrays comprised of large numbers of immobilized protein capture agents. While most efforts in this arena have focused on the use of biomolecules such as antibodies and nucleic acid aptamers as capture agents, synthetic species have many potential advantages. However, synthetic molecules isolated from combinatorial libraries generally do not bind target proteins with the high affinity necessary for array applications. Here, we demonstrate that simple linear peptides bind dimeric proteins tenaciously when immobilized, although they exhibit only modest affinity in solution. These data show that high-affinity bidentate capture agents for dimeric proteins can be created by simply immobilizing modest-affinity ligands on a surface at high density, bypassing the requirement for careful optimization of linker length and geometry that is normally required to create a high-affinity solution bidentate ligand.

Combinatorial Chemistry Techniques↗