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High throughput PCR detection of HCV based on semiautomated multisample RNA capture.

An amplification procedure based on a semiautomated 60-sample RNA capture, including combined reverse transcription/polymerase chain reaction (RT-PCR) and nested PCR/Taqman amplicon detection. is described. It can be completed within a working day and is suitable for the development of a fully automated system. HCV RNA-specific capture is independent of the sequence variations as it targets the poly(U) tract commonly present at the 3'-end of the HCV genome (U-capture). The type specificity of the assay determined in a panel of 56 confirmed HCV antibody-positive samples (genotypes 1-6) was slightly better when compared to a commercial assay. The sensitivity evaluated on serial dilutions of representative samples was equal for genotypes 1, 2, 5, 6, or increased for genotypes 3 and 4 with the U-capture assay.

Automation↗

Ease of capture in lines of Japanese quail (Coturnix japonica) subjected to contrasting selection for fear or sociability.

Japanese quail (Coturnix japonica) of lines, which have been subjected to contrasting selection for duration of the tonic immobility (TI) reaction or social reinstatement (SR) behaviour over many generations show corresponding differences in underlying fearfulness and sociality. As fearfulness and sociality are particularly influential traits in domesticated species, the finding that such traits respond to artificial selection may have important implications for poultry welfare and performance. However, it is not known if or how such selection has influenced human-animal interactions. The present experiment investigated the influence of fearfulness and SR behaviour on the ease with which birds could be caught and handled. Birds of lines selected for duration of the TI response or SR behaviour were reared in mixed line groups (LTI and STI or HSR and LSR) of 491 and 346 birds, respectively, until 6 weeks of age. When the birds were 2, 4, and 6 weeks of age, they were caught one by one and their individual capture ranks noted. In the group of birds selected for duration of the TI response, birds selected of the line selected for short duration of TI were caught before those selected for long duration of the response. In the group of birds selected for SR motivation, birds of the high line were caught before their low lines counterparts. Coefficients of concordance between capture ranks were significant and capture ranks did not differ significantly across ages. These results imply that selection for low levels of fear or high levels of sociality produces animals that are less disturbed by human interventions than animals selected for the opposite traits. The greater ease of capture of low fear line birds than high fear line birds may be explained by reduced fear of humans. The fact that the birds selected for high levels of SR behaviour are easier to catch than birds selected for low levels of sociality is less readily explicable. One hypothesis is that HSR line chicks tend to be more strongly imprinted on each other and the human caretaker. However, SR behaviour is highly species specific in both lines, existing evidence for line differences in social discrimination is limited and birds of the two lines show similar duration of the TI response. Despite this, whatever their underlying causation, these results demonstrate that genetic selection can be used to reduce negative reactions to human beings and may be of value in the improvement of both animal welfare and productivity.

Journal Article↗

[Incidence of pulmonary tuberculosis: application of the capture-recapture method].

OBJECTIVE: To determine as accurately as possible the incidence of pulmonary tuberculosis within Health Area 15 of the Community of Valencia during the period 1990-1993, using the capture-recapture method. METHOD: Descriptive study on the population of Health Area 15 (population: -139.903) divided into 4 large groups according to age (0-14, 15-34, 34-54 and 55+). Data was obtained from the statutory notification system of infectious disease (SNSID) and from the registry of the said area's Hospital Microbiology SERVICE: The main variable under study was the number of cases of pulmonary tuberculosis, both as notified under the SNSID and in terms of cases in the microbiology register in which M. tuberculosis was isolated. The incidence-rates were calculated by age and year of study for both registries employing the capture-recapture method. RESULTS: The mean annual incidence obtained for the SNSID register during the study period was 15.85 cases x 10(-5), and for the microbiology registry it was 23.29 x 10(-5). When the capture-recapture method was employed the mean annual incidence for the study period was 34.81 x 10(-5) (CI 95%: 31.82-39.92). In each of the years studied the number of cases identified was greater for the microbiology register than for the SNSID. Around half the cases of tuberculosis are below 34 years of age, with the larger section, and that having the highest incidence of tuberculosis being the 15 to 34 years age-group. There is no apparent upward trend in incidence rates calculated for this period. CONCLUSIONS: The data from the SNSID system on incidence of pulmonary tuberculosis within Health Area 15 of the Community of Valencia tends to underestimate the true incidence rate. The factual resources of the Hospital Microbiology Service are underutilized, considering the quantity and quality of information it can provide. The capture-recapture method is a good choice of method for measuring tuberculosis incidence. This method merits greater use within the field of epidemiology as much in order to assess the representativeness and thoroughness of surveillance systems as to identify inadequacies in their reporting and localisation of disease outbreaks.

Adolescent↗

A vision-based three-dimensional capture system for maxillofacial assessment and surgical planning.

We describe a vision-based three-dimensional facial data capture system designed for the planning of maxillofacial operations. We describe the system requirements and outline the methods used to develop a complete three-dimensional facial capture system. Our approach is based upon imaging the face using two stereo-pair sets of cameras. Scale-space-based stereo-matching is then used to recover correspondences between each of the captured stereo-pairs. Photogrammetric routines based on adjustment of bundles are used off-line to calibrate the system by imaging a single object that references all cameras to the same co-ordinate frame. This calibration scheme allows us to convert stereo correspondences to world points for each pair of cameras without the need for any subsequent fusion of data. Initial results show that we are able to capture key facial landmarks to within 0.5 mm.

Algorithms↗

Capture ELISA systems for the detection of bovine coronavirus-specific IgA and IgM antibodies in milk and serum.

Isotype-capture ELISAs for BCV-specific IgA and IgM were developed and tested on milk and serum samples from Swedish cattle. The capture ELISAs showed higher sensitivity than indirect ELISAs for detection of BCV-specific IgA and IgM. In the capture ELISAs the agreement between detection in milk and serum samples was 94% for IgA and 86% for IgM. The correlation between log(10) titres in milk and serum was r=0.82 (P<0.001) for IgA and 0.84 (P<0.001) for IgM. Milk seemed a better target than serum for diagnosing specific IgA at low levels. There was no variation in the isotype-specific BCV antibody titres between healthy quarters of the same udder, but subclinical mastitis was associated with higher levels of IgA antibodies and weak false IgM positive reactions in undiluted milk. Bovine IgA and IgM antibodies in milk and serum showed high stability towards freezing and thawing and storage at room temperature. The antibody responses to BCV were followed in milk and serum from six dairy cows and in serum from four calves for a period of 1 year after an outbreak of winter dysentery (WD). In this outbreak some animals became reinfected with BCV. The IgA and IgM capture ELISAs differentiated between primarily BCV infected and reinfected animals. In the primarily infected cattle, IgM antibodies were first detected in milk and serum four to nine days after the first WD symptoms observed, and were subsequently detected for at least 2-3 weeks. IgM was also detected in the reinfected cows, but mostly at lower levels and for a shorter period of time than in the primarily infected animals. In milk, however, the IgM response of the reinfected cows was detected for a longer period of time than in serum. Six months after the outbreak, IgA was still detected in both serum and milk of all six cows and also in serum of one calf. The reinfected cows showed higher and more long-lasting peak levels of IgA in milk and serum than the primarily infected cows, indicating boosting of the IgA response.

Animals↗

Noninvasive repositioning of a posterior chamber intraocular lens following pupillary capture.

A case of pupillary capture of a posterior chamber intraocular lens (IOL) is presented. A nonsurgical approach that may help reposition an IOL optic entrapped in a pupillary capture when chemical mydriasis fails is described. Suggestions for a logical progression of nonsurgical procedures to help release optic capture are presented. Possible causes and preventative measures to decrease the incidence of pupillary capture of a posterior chamber IOL are discussed.

Adult↗

Incidence, causes, and neodymium:YAG laser treatment of pupillary capture.

OBJECTIVE: To report the incidence of and factors associated with pupillary capture after cataract surgery and evaluate the outcomes of treatment with the neodymium:YAG (Nd:YAG) laser. SETTING: Unit of Ophthalmology, University of Edinburgh, and Royal Infirmary of Edinburgh Trust Hospital, Edinburgh, United Kingdom. METHODS: This prospective study comprised 792 consecutive patients having cataract extraction by one surgeon from 1989 to 1993. Outcome measures were incidence of pupillary capture associated with eye disease and treatment, surgical technique, and intraocular lens (IOL) placement and style; postoperative progress; and results of Nd:YAG treatment. RESULTS: Pupillary capture developed in 30 of the 764 patients (3.9%) having implantation of a posterior chamber IOL a mean of 14 weeks postoperatively (range 2 to 44 weeks). The incidence was significantly higher in eyes with angle-closure glaucoma, combined glaucoma and cataract surgery, can-opener capsulotomy, manual extraction, sulcus IOL implantation, large-optic IOLs, and one-piece IOLs. Neodymium:YAG laser treatment was successful in 8 of 12 eyes. CONCLUSION: Pupiliary capture may be anticipated and when recognized at an early stage, treated successfully with an Nd:YAG laser in most cases.

Administration, Topical↗

Pupillary block after pupillary capture of an AcrySof intraocular lens.

A 54-year-old man developed pupillary block resulting from pupillary capture 2 months after uneventful phacomulsification and AcrySof intraocular lens (IOL) implantation. The IOL was placed in the bag through a 6.0 mm continuous curvilinear capsulorhexis. The glaucoma was treated with intravenous drip infusion of a hyperosmotic diuretic, followed by peripheral iridectomy, iridocapsular synechiolysis, and IOL repositioning. The IOL loops were located completely in the bag. However, pupillary capture recurred 3 weeks after the surgery, at which time intraocular pressure was normal. The flexibility of the IOL optic and its large overall length and rigid, low-angulated loops were the probable causes for the recurrence of the pupillary capture. The IOL was exchanged for a sulcus-fixated, single-piece poly(methylmethacrylate) lens with 10 degree angulated loops. Pupillary capture did not recur during the follow-up.

Acrylic Resins↗

Small incision nucleus capture: results of 200 cases.

PURPOSE: To compare the learning curve in a series of 200 cataract surgeries performed using small incision nucleus capture with that of phacoemulsification as reported in the literature. SETTING: Department of Ophthalmology, University of Genoa, Genoa, Italy. METHODS: Two hundred eyes of 163 consecutive patients with cataract had small incision nucleus capture, a relatively new cataract surgery technique that allows small incisions and in-the-bag intraocular lens implantation. Patients were divided into 4 groups of 50 each according to when they had surgery between August 1996 and October 1997. The incidence of intraoperative complications (capsule break with or without vitreous loss, capsulorhexis tears, Descemet's detachment, transient iris damage) and postoperative complications (raised intraocular pressure, corneal epithelial edema, Descemet's folds, and permanent iris damage) were evaluated at the different time points. Also recorded was final visual acuity. These results were compared with those obtained with phacoemulsification. RESULTS: The study comprised 92 women and 71 men with an age range of 41 to 93 years. Overall final results showed that the learning curve of nucleus capture is comparable to that of phacoemulsification. CONCLUSION: Nucleus capture cataract extraction resulted in a low incidence of complications and good visual recovery that was comparable to that obtained with phacoemulsification.

Adult↗

Borehole parametric study for neutron induced capture gamma-ray spectrometry using the MCNP code.

The MCNP Monte Carlo code has been used to simulate neutron transport from an Am-Be source into a granite formation surrounding a borehole. The effects of the moisture and the neutron poison on the thermal neutron flux distribution and the capture by the absorbing elements has been calculated. Thermal and nonthermal captures for certain absorbers having resonance structures in the epithermal and fast energy regions such as W and Si were performed. It is shown that for those absorbers having large resonances in the epithermal regions when they are present in dry formation or when accompanied by neutron poisons the resonance captures may be significant compared to the thermal captures.

Journal Article↗

In vitro model for the evaluation of inferior vena cava filters: effect of experimental parameters on thrombus-capturing efficacy of the Vena Tech-LGM filter.

PURPOSE: To determine the experimental parameters in an in vitro model that influence the thrombus-capturing efficacy of the Vena Tech-LGM filter. MATERIALS AND METHODS: The Vena Tech-LGM filter was evaluated in an in vitro model of the vena cava with a computer-controlled flow system with a total of 5,200 thrombi. The influences of the following experimental parameters on the capture rate were analyzed with a multiple logistic regression model: type of testing (single, double, and multiple shot testing), thrombus diameter and length, IVC diameter and orientation, flow quality and quantity, flow velocity, and the length of the prepositioned thrombus. RESULTS: A significant influence on the capture rate could be demonstrated for the type of testing, the thrombus diameter and length, the IVC diameter, and with double shot testing for the length of the prepositioned thrombus and the IVC orientation. The flow quality and the peak velocity were not significant. Based on these results, a protocol for in vitro testing of IVC filters was designed. CONCLUSIONS: Experimental parameters influence the thrombus-capturing efficacy of the Vena Tech-LGM filter and should be taken into account when in vitro testing is performed.

Filtration↗

The use of digital image capture in contact lens practice.

The present decade has seen the introduction of digital imaging systems to a number of medical environments, including contact lens practice. During the past year, the authors have used two anterior segment video-based digital image capture systems. These systems allow instantaneous image capture and have both diagnostic value to practitioners and educational value to patients. This paper discusses and clarifies the terminology that surrounds such systems and looks carefully at the advantages and disadvantages when compared to more traditional 35 mm photography in particular with reference to image colour, resolution and storage. The difference between video digital image capture and the more recently available digital camera image capture is also discussed. The value of digital imaging is shown by a series of images from the authors' archives.

Journal Article↗

Endostatin capture from Pichia pastoris culture in a fluidized bed. From on-chip process optimization to application.

One of the characteristics of the methylothrophic yeast Pichia pastoris is its ability to grow to a very high cell density. Biomass concentrations of 300-400 g wet mass/l are common. It is therefore obvious that the recovery processes of extracellular proteins from this microorganism should take into account the effect of high biomass content. Separation by filtration and/or centrifugation is possible but these steps are cumbersome and can affect the protein recovery. The use of fluidized beds is attractive proteins capture option since it eliminates the biomass while capturing the desired protein. Zirconia-based resins possess unique properties which make them appropriate for processing high biomass concentrations in an expanded bed mode. The beads are particularly heavy (density is 3.2 g/ml) and small (75 microm) and therefore can accommodate high fluidization velocity and high mass transport. Specific operating conditions for effective capture of expressed protein have to be determined. This determination is generally time consuming and requires relatively large amount of feedstock for the lab trials. To avoid multiple chromatographic trials in columns, optimal conditions of adsorption and elution were determined by ProteinChip technology coupled with mass spectrometry. This technology involves flat chip surfaces functionalized as chromatographic beads where it is possible to adsorb and desorb proteins. Four different functional groups (strong anion-exchange, weak cation-exchange, hydrophobic and metal chelate) were tested and the retained proteins were analyzed directly by mass spectrometry. The weak cation-exchange group was chosen for further work. The Zirconia-based weak cation-exchange sorbent (CM HyperZ) was evaluated for binding capacity in a packed column and then for capturing endostatin from crude feed stock. Based on the previously determined conditions; 45 l of culture containing approximately 15 kg of biomass (wet mass) and 3 g endostatin were applied on an expanded bed at a flow-rate of 535 cm/h, yielding 80% of the endostatin and removing approximately 80% of foreign proteins.

Cation Exchange Resins↗

A micro-capture ELISA for detecting Mycoplasma pneumoniae IgM: comparison with indirect immunofluorescence and indirect ELISA.

A mu-capture ELISA was developed for detecting Mycoplasma pneumoniae-specific IgM, and compared with an indirect immunofluorescent antibody (IFA) technique and an indirect ELISA. mu-capture ELISA and IFA compared well and were found to be the most sensitive assays. The IFA test can be completed in 2 h whilst the results of the mu-capture ELISA can be available in 24 h. Both tests are amenable to routine diagnostic use and have similar sensitivity. Indirect ELISA was found to be less sensitive and less specific, giving high assay values with several sera having undetectable M. pneumoniae CF antibody or CF antibody in low titre. Serum samples obtained from 11 patients at various times after M. pneumoniae infection showed maximum antibody levels within the first month by all assays, with a gradual fall in amount of IgM with time when assayed by mu-capture ELISA, a more gradual decline by IFA and hardly any decline with indirect ELISA. It was concluded that the indirect ELISA is unsuitable for the investigation of possible M. pneumoniae infection because the sustained high assay values with serum samples taken many months after infection, make interpretation of the test results very difficult.

Antibodies, Bacterial↗

Completeness of notification of tuberculosis in The Netherlands: how reliable is record-linkage and capture-recapture analysis?

The aim of this study was to describe a systematic process of record-linkage, cross-validation, case-ascertainment and capture-recapture analysis to assess the quality of tuberculosis registers and to estimate the completeness of notification of incident tuberculosis cases in The Netherlands in 1998. After record-linkage and cross-validation 1499 tuberculosis patients were identified, of whom 1298 were notified, resulting in an observed under-notification of 13.4%. After adjustment for possible imperfect record-linkage and remaining false-positive hospital cases observed under-notification was 7.3%. Log-linear capture-recapture analysis initially estimated a total number of 2053 (95% CI 1871-2443) tuberculosis cases, resulting in an estimated under-notification of 36.8%. After adjustment for possible imperfect record-linkage and remaining false-positive hospital cases various capture-recapture models estimated under-notification at 13.6%. One of the reasons for the higher than expected estimated under-notification in a country with a well-organized system of tuberculosis control might be that some tuberculosis cases, e.g. extrapulmonary tuberculosis, are managed by clinicians less familiar with notification of infectious diseases. This study demonstrates the possible impact of violation of assumptions underlying capture-recapture analysis, especially the perfect record-linkage, perfect positive predictive value and absent three-way interaction assumptions.

Disease Notification↗

Primary resource capture in two sympatric Armillaria species in managed Norway spruce forests.

The ability of the preferentially saprotrophic fungus Armillaria cepistipes and the pathogenic A. ostoyae to capture fresh stump resources was investigated in managed Alpine Norway spruce (Picea abies) forests where both species occur sympatrically. The incidence of Armillaria species and genets as rhizomorphs in the soil as well as external and internal (heart rot) stump colonisation were determined in three comparable 1 ha plots. The results indicate that A. cepistipes and A. ostoyae have a very similar strategy to capture fresh stump resources. Both species produce dense networks of rhizomorphs in the soil enveloping living trees. After the felling of the trees, the stumps are rapidly captured through spread of mycelial fans in the cambial zone. Our study suggests that A. ostoyae is slightly more competitive than A. cepistipes in primary resource capture.

Agaricales↗

A resonant electron capture time-of-flight MS with trochoidal electron monochromator.

A prototype electron monochromator (EM) reflectron time-of-flight (TOF) mass spectrometer has been constructed and demonstrated to record resonant electron capture (REC) mass spectra of electron-capturing compounds. The electron energy is ramped from -1.7 to +25 eV at a preset frequency, and the energy spread of the electron beam at 15 nA is 100 meV or better. Ions are orthogonally extracted into the analyzer at a frequency of up to 80 kHz while maintaining an upper m/z-limit of at least 300 and a mass resolving power of approximately 1000. A complete REC mass spectrum, which includes an effective yield versus electron energy curve for each negative ion formed from the compound being analyzed, typically takes several days to produce with a quadrupole or magnetic sector mass spectrometer. With the EM TOF described in this work, three-dimensional negative ion electron capture spectra are recorded in an interval on the order of only 1 s and displayed in real time. This new analytical capability could make it possible to perform GC REC mass spectrometry as well as easier (a) to measure the temperature dependence of REC cross sections, (b) to determine enthalpies of negative ion formation (accurate determination of the enthalpy of ion formation requires knowledge of the translational energy released during a dissociative capture event), and (c) to provide complete thermochemical descriptions of dissociative electron attachment by measuring ion lifetimes.

Anions↗

Analysis for TNF-alpha using solid-phase affinity capture with radiolabel and MALDI-MS detection.

Screening of mutant mice for subtle phenotypes requires sensitive, high-throughput analyses of sentinel proteins in functional pathways. The cytokine TNF-alpha is upregulated during inflammatory reactions associated with autoimmune diseases. We have developed a method to monitor the concentration of TNF-alpha under physiological conditions. TNF-alpha is captured, purified, and concentrated using monoclonal antibody-coated microbeads. The capture is efficient (> 80%) and can be used in the concentration range < 100 pg/mL to > 50 ng/mL, as determined by detection of 125I-labeled TNF-alpha. The bead capture of TNF-alpha can be combined with direct detection by MALDI-MS for sample concentrations of > 10 ng/mL. TNF-alpha can be captured and detected from diluted mouse serum, with minimal interferences observed in the MALDI spectrum. This method is adaptable to high-throughput sample handling with microfluidic devices and automated mass spectrometric analysis.

Animals↗