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AIDS vaccine: Intranasal immunization using inactivated HIV-1-capturing core-corona type polymeric nanospheres.

Polymeric nanospheres have been widely used in biomedical applications, such as drug, gene and vaccine delivery systems. Nanospheres with entrapped antigens have recently been shown to possess significant potential as vaccine delivery systems and adjuvants. We previously reported that concanavalin A-immobilized polystyrene nanospheres (Con A-NS) could efficiently capture HIV-1 particles and intranasal immunization with inactivated HIV-1-capturing nanospheres (HIV-NS) induced vaginal anti-HIV-1 IgA antibody responses in mice. In addition, vaginal washes from intranasally immunized mice were capable of neutralizing HIV-1. Moreover, simian/human immunodeficiency virus KU-2-capturing nanospheres (SHIV-NS) immunized macaques exhibited partial protection when vaginally and systemically challenged with pathogenic viruses. HIV-NS is suggested to be particularly suitable to enhance antigen delivery to dendritic cells (DCs). In this study, we investigated the mucosal antibody response in mice after the intravaginal or intranasal immunization in detail with using different sized (360, 660, 940 and 1230 nm) HIV-NS. The amount of immobilized Con A to NS was dependent on the surface area of the particle. Moreover, Con A-NS with different sizes could equally capture inactivated HIV-1. Intravaginal or intranasal immunization by HIV-NS with diameters ranging 360 to 1230 nm significantly induced vaginal antibody responses. However, significant differences on vaginal anti-HIV-1 gp120 IgA and IgG antibodies were not found after intravaginal or intranasal immunization with different sized HIV-NS. These results suggest that HIV-NS provides an efficient vaccine delivery system for the induction of a mucosal immune response and the development of a mucosal vaccine.

AIDS Vaccines↗

Evaluation of a commercial dengue NS1 antigen-capture ELISA for laboratory diagnosis of acute dengue virus infection.

A commercial dengue NS1 antigen-capture ELISA was evaluated to demonstrate its potential application for early laboratory diagnosis of acute dengue virus infection. Dengue virus NS1 antigen was detected in 199 of 213 acute serum samples from patients with laboratory confirmation of acute dengue virus infection but none of the 354 healthy blood donors' serum specimens. The dengue NS1 antigen-capture ELISA gave an overall sensitivity of 93.4% (199/213) and a specificity of 100% (354/354). The sensitivity was significantly higher in acute primary dengue (97.3%) than in acute secondary dengue (70.0%). The positive predictive value of the dengue NS1 antigen-capture ELISA was 100% and negative predictive value was 97.3%. Comparatively, virus isolation gave an overall positive isolation rate of 68.1% with a positive isolation rate of 73.9 and 31.0% for acute primary dengue and acute secondary dengue, respectively. Molecular detection of dengue RNA by RT-PCR gave an overall positive detection rate of 66.7% with a detection rate of 65.2 and 75.9% for acute primary dengue and acute secondary dengue, respectively. The results indicate that the commercial dengue NS1 antigen-capture ELISA may be superior to virus isolation and RT-PCR for the laboratory diagnosis of acute dengue infection based on a single serum sample.

Acute Disease↗

Ambient odors modulate visual attentional capture.

Sudden visual events capture attention involuntarily because they may signal potential threats. Some theoretical accounts consider that the biological significance of these events is established through the limbic structures. Thus, the manipulation of the limbic activity would affect attentional capture. Since these structures are directly linked to the olfactory system, we have tended to modulate their activity with olfactory stimulations. We have examined behavioral performance in a task of attentional capture by luminance under conditions of ambient odors. Our results show that attentional capture is indeed modulated by ambient odors, and that this modulation may depend on the odor's properties.

Adult↗

A Bayesian, combinatorial approach to capture-recapture.

It is shown that, in the capture-recapture method, the widely used formulae of Bailey or Chapman-Seber give the most likely value for the size of the population, but systematically underestimate the probability that the population is larger than any given size. We take here a first step in a combinatorial approach which does not suffer from this flaw: formulae are given which can be used in the closed case (no birth, death or migrations between captures) when at least two animals have been recaptured and when there is homogeneity with regard to capture probability. Numerical and heuristic evidence is presented pointing to the fact that the error incurred when using the formulae of Bailey or Chapman-Seber depends asymptotically only on the number of recaptured animals, and will not diminish if the number of captured animals becomes large while the number of recaptured animals remains constant. A result that was stated and left unproven by Darroch is proven here.

Analysis of Variance↗

Determination of rat hepatic polyamines by electron-capture gas chromatography.

INTRODUCTION: An efficient and reproducible electron-capture gas chromatographic protocol that allows the simultaneous detection and quantification of the polyamines putrescine (1,4-diaminobutane), cadaverine (1,5-diaminopentane), and spermidine (N-[3-aminopropyl]-1,4-diaminobutane) was developed. METHODS: Hepatic tissue from male Sprague-Dawley rats was used for analysis. The polyamines and the internal standard (sertraline) were extracted and derivatized with pentafluorobenzoyl chloride (PFBC) under basic aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column (narrow-bore fused silica column; 25 mm x 0.32 mm) and an electron-capture detector. RESULTS: PFBC reacts with the amine functions of the polyamines examined here to produce PFB derivatives with high sensitivity on electron-capture detection. The method permitted the quantitative analyses of all three amines in rat hepatic tissue; the concentration of putrescine, but not spermidine, was increased significantly following a 14-day administration of the diamine oxidase (DAO) inhibitor aminoguanidine. Cadaverine was also present at increased concentrations in hepatic homogenates from aminoguanidine-treated rats. DISCUSSION: Extractive derivatization with PFBC followed by gas chromatographic analysis using electron-capture detection results in a rapid and reproducible assay that permits the simultaneous detection and quantification of putrescine, cadaverine, and spermidine in biological tissue.

Animals↗

Prey capture behavior and kinematics of the Atlantic cownose ray, Rhinoptera bonasus.

The structurally reinforced jaws of the cownose ray, Rhinoptera bonasus testify to this species' durophagous diet of mollusks, but seem ill-suited to the behaviors necessary for excavating such prey. This study explores this discordance by investigating the prey excavation and capture kinematics of R. bonasus. Based on the basal suction feeding mechanism in this group of fishes, we hypothesized a hydraulic method of excavation. As expected, prey capture kinematics of R. bonasus show marked differences relative to other elasmobranchs, relating to prey excavation and use of the cephalic lobes (modified anterior pectoral fin extensions unique to derived myliobatiform rays). Prey are excavated by repeated opening and closing of the jaws to fluidize surrounding sand. The food item is then enclosed laterally by the depressed cephalic lobes, which transport it toward the mouth for ingestion by inertial suction. Unlike in most sharks, upper jaw protrusion and mandibular depression are simultaneous. During food capture, the ray's spiracle, mouth, and gill slit movements are timed such that water enters only the mouth (e.g., the spiracle closes prior to prey capture and reopens immediately following). Indigestible parts are then hydraulically winnowed from edible prey portions, by mouth movements similar to those used in excavation, and ejected through the mouth. The unique sensory/manipulatory capabilities of the cephalic lobes, as well as the cownose ray's hydraulic excavation/winnowing behaviors and suction feeding, make this species an effective benthic predator, despite its epibenthic lifestyle.

Analysis of Variance↗

The effects of capture spiral composition and orb-web orientation on prey interception.

Cribellar prey capture threads found in primitive, horizontal orb-webs reflect more light, including ultraviolet wavelengths, than viscous threads found in more derived, vertical orb-webs. Low web visibility and vertical orientation are each thought to increase prey interception and may represent key innovations that contributed to the greater diversity of modern, araneoid orb-weaving spiders. This study compares prey interception rates of cribellate orb-webs constructed by Uloborus glomosus (Uloboridae) with viscous orb-webs constructed by Leucauge venusta (Tetragnathidae) and Micrathena gracilis (Araneidae). We placed sectors of cribellar and viscous threads side by side in frames that were oriented either horizontally or vertically. The webs of both U. glomosus and L. venusta intercepted more prey when vertically oriented. In each orientation L. venusta webs intercepted more insects than did U. glomosus. Although this is consistent with the greater visibility of cribellar threads, the more closely spaced capture spirals of L. venusta may have contributed to this difference. Micrathena gracilis webs intercepted more prey than did U. glomosus webs, although web orientation did not affect the performance of this araneoid species. The stickier and more closely spaced capture spirals of M. gracilis may have enhanced the interception rates of this species and accounted for the greater number of smaller dipterans retained in its webs. The tendency for these slow, weak flight insects to be blown into both horizontal and vertical webs may account for similar interception rates of horizontal and vertical M. gracilis webs. These observations support the enhanced prey interception of vertically oriented orb-webs, but offer only qualified support for the contributions of lower visibility viscous capture threads.

Animals↗

Age-related effects of attentional and oculomotor capture by onsets and color singletons as a function of experience.

The present experiment examined the degree to which experience with different stimulus characteristics affects attentional capture, particularly as related to aging. Participants were presented with onset target/color singleton distractor or color singleton target/onset distractor pairs across three experimental sessions. The target/distractor pairs were reversed in the second session such that the target in the first session became the distractor in the second and third sessions. For both young and old adults previous experience with color as a target defining feature influenced oculomotor capture with task-irrelevant color distractors. Experience with sudden onsets had the same effect for younger and older adults, although capture effects were substantially larger for onset than for color distractors. Experience-based capture effects diminished relatively rapidly after target and distractor-defining properties were reversed. The results are discussed in terms of top-down and stimulus-driven effects on age-related differences in attentional control.

Adult↗

Capture and reconstitution of G protein-coupled receptors on a biosensor surface.

Surface plasmon resonance (SPR) biosensors offer a unique opportunity to study the binding activity of G protein-coupled receptors (GPCRs) in real time with minimal sample preparation. Using two chemokine receptors (CXCR4 and CCR5) as model systems, we captured the proteins from crude cell preparations onto the biosensor surface and reconstituted a lipid environment to maintain receptor activity. The conformational states of the receptors were probed using conformationally dependent antibodies, and by characterizing the binding properties of a native chemokine ligand (stromal cell-derived factor 1alpha). The results suggest that the detergent-solubilized receptors are active for ligand binding in the presence and absence of a reconstituted bilayer. There are three advantages to using this receptor-capturing approach: (1) there is no need to purify the receptor prior to immobilization on the biosensor surface, (2) the receptors are homogeneously immobilized through the capturing step, and (3) the receptors can be captured at high enough densities to allow the study of relatively low-molecular-mass ligands (2000-4000Da). We also demonstrated that the receptors are sensitive to the solubilizing conditions, which illustrates the potential for using SPR biosensors to rapidly screen solublization conditions for GPCRs.

Antibodies, Monoclonal↗

Comparison of electron-capture and electrolytic conductivity detection for the gas-liquid chromatographic analysis of some perfluoro derivatives of four agricultural chemicals.

The gas chromatographic response of four perfluoro derivatives, of four agricultural chemical [diethylstilbestrol] (DES) clopidol, linuron and carbofuran] was examined. The derivatives varied in fluorine content from 3 to 30 atoms per molecule. The sensitivities of the derivatives were found to be ca. 10-100 times greater by electron-capture detection than by electrolytic conductivity detection (halogen mode). The sensitivity also was found to increase with increasing fluorine content for all derivatives by electrolytic conductivity, whereas by electron capture, DES and clopidol exhibited similar responses with either the penta-, hepta-, or pentadeca-fluoro derivatives. The sensitivity of linuron and carbofuran derivatives by electron capture varied with increasing fluorine content. For detection by electron capture, the hepta-fluorobutyryl derivatives were preferred for DES and clopidol, and either the hepta-fluorobutyryl or the pentadecafluorooctanoyl derivatives for linuron and carbofuran. For detection by electrolytic conductivity, the pentadecafluorooctanoyl derivatives were superior for all four compounds.

Carbofuran↗

A generic capture ELISA for recombinant proteins fused to glutathione S-transferase: validation for HPV serology.

An enzyme-linked immunosorbent assay (ELISA) system has been developed that uses glutathione crosslinked to casein as capture protein to bind recombinant protein antigens fused to N-terminal glutathione S-transferase (GST). The method allows simple and efficient immobilization and one-step purification of overexpressed recombinant antigens from crude lysates on ELISA plates coated with glutathione casein. Several antigens can be tested in parallel under the same conditions without the need to biochemically purify or renature the proteins. An additional undecapeptide epitope fused to the C-terminus of each antigen permits the detection and quantification of any full-length protein antigen bound to the ELISA plate with one single monoclonal antibody. The ELISA system was applied with four antigens to detect antibodies against E6 and E7 proteins of human papillomavirus types 16 and 18. Antibody reactivities of 164 sera from patients with cervical carcinoma and healthy individuals were in good agreement with those determined using a previously established capture ELISA with biochemically purified and renatured proteins as antigens although the GST capture ELISA was more sensitive with no loss of specificity. The GST capture ELISA could be adapted to provide standardized antibody assays for many protein antigens.

Antibodies, Viral↗

Factors influencing the measurement of oestrone sulphate by dipstick particle capture immunoassay.

Factors influencing the performance of a dipstick competitive particle capture immunoassay (PCI) for the steroid oestrone sulphate (OS) were investigated. Appropriate 'blocking' of the nitrocellulose dipstick membrane was necessary for the upward flow of microsphere particles. Traditional protein blocking agents including BSA, gelatin and casein were unsatisfactory while synthetic polymers and surfactants were effective in promoting microsphere movement. A simple buffer consisting of 1% aqueous NaCl containing 0.05% Tween 20 was suitable for carrying the components up the dipstick and facilitating the antibody-antigen interactions. Increasing microsphere diameters from 0.3 to 0.8 microm allowed the microsphere antibody coating concentration to be reduced which enabled lower concentrations of OS to be measured. However, upward flow rate and the maximum signal attainable was compromised as a consequence. Enlarging the dipstick membrane nominal pore size from 3 to 12 microm increased the speed of test dot development, but assay sensitivity suffered as a result in some instances. Changing the capture antigen markedly influenced the dose-response lines. No dose-response was achieved with OG-BSA as the capture antigen while OHS-BSA and OCMO-BSA as capture antigens produced dose-response lines with means +/- S.E.M. EC(50) values of 140 +/- 16 and 19 +/- 1 ng/ml, respectively.

Antigens↗

Capture-ELISA based on recombinant PR3 is sensitive for PR3-ANCA testing and allows detection of PR3 and PR3-ANCA/PR3 immunecomplexes.

Proteinase 3 (PR3), a constituent of azurophil granules of neutrophils (polymorphonuclear cells, PMNs), is the target antigen for most anti-neutrophil cytoplasmic antibodies (c-ANCA) in Wegener's granulomatosis (WG). We have recently developed an expression system for recombinant PR3 (rPR3) that is recognized by c-ANCA. Here, we report on the development and characterization of two monoclonal antibodies (moABs) and a rabbit polyclonal antiserum generated against this rPR3. Epitope competition analysis indicates that the moABs MCPR3-1 and MCPR3-2 recognize overlapping epitopes on the PR3 molecule that are distinct from the ones recognized by moABs 4A5 and 6A6 developed by others. Since MCPR3-2 does not appear to compete for epitopes recognized by a sizable proportion of PR3-ANCA, we used it to develop a sensitive capture enzyme linked immunosorbent assay (ELISA) for clinical PR3-ANCA testing. Both purified PMN PR3 and crude human mast cell line (HMC-1)/PR3-S176A cell lysates were used as sources of PR3 target antigen in this assay with equal analytical sensitivity and specificity. Of 109 patients with ANCA-associated disease, 91 (83.5%) and 90 (82.6%) were PR3-ANCA positive by capture ELISA when PMN-PR3 and HMC-1/PR3-S176A cell lysates were used as antigen, respectively. When HMC-1/PR3 and HMC-1/PR3-S176A cells were used as indirect immunofluorescence (IIF) substrate, 88 (80.7%) and 92 (84.4%) were PR3-ANCA positive, respectively. These differences were not statistically significant. Only 1 of 151 controls without defined ANCA-associated disease tested positive by capture ELISA with either target antigen (both negative by PR3-ANCA specific IIF). The capture ELISA can also be used to detect of PR3-ANCA immunecomplexes and, in combination with the rabbit antiserum, for the quantitative measurement of PR3 in biological fluids.

Animals↗

Flux ratio theorems for nonstationary membrane transport with temporary capture of tracer.

It has been shown recently that the ratio of unidirectional tracer fluxes, passing in opposite directions through a membrane which has transport properties varying arbitrarily with the distance from a boundary, is independent of time from the very first appearance of the two outfluxes from the membrane. This surprising proposition has been proved for boundary conditions defining standard unidirectional fluxes, and then generalized to classes of time-dependent boundary conditions. The operational meaning of all the resulting theorems is that when any of them appear to be refuted experimentally, the presence of more than one parallel transport pathway (that is, of membrane heterogeneity transverse to the direction of transport) can be inferred and analyzed. Recent experimental data have been interpreted accordingly. However, the proofs of the theorems given so far have not taken into account the possibility of temporary capture of tracer at sites fixed in the membrane (including also entrances to microscopic culs-de-sac). The possible presence of such a process, which would not affect fluxes in the steady state, left a fundamental gap in the aforementioned inferences. It is shown here that all the theorems previously proved for the flux ratio under unsteady conditions remain valid when temporary capture of tracer is admitted, no matter how the rate of capture, and the probability distribution of residence times of tracer at capture sites, may depend on the distance from a membrane boundary. The validity of the aforementioned inferences from observed time-dependence of the flux ratio is thereby extended to a much wider class of membrane transport processes.

Biological Transport↗

Color, form and luminance capture attention in visual search.

Extant models of visual attention predict that a salient element should produce a bottom-up activation leading to a stimulus-driven attentional capture (e.g. Cave, 1999). However, apart from onset, previous works manipulating set-size in visual search failed to provide empirical evidence for this kind of capture. By varying target-singelton distance method, based on a single set-size, we explored whether, in a serial search task, an attentional capture is triggered by static discontinuities such as those generated through the manipulation of color, form, and luminance. The results suggest that those physical properties are indeed able to capture attention automatically.

Adult↗

Stereo capture: local rematching driven by binocularly attended 3-D configuration rather than retinal images.

Previous explanations for stereo capture were mainly based on the low-level perceptual processing of binocular stereopsis which usually shows that one pair of retinal images corresponds to only one 3-D perceptual configuration. Stereo capture, however, may encounter multiple perceptual configurations due to the matching ambiguity of wallpaper elements that may not be solved merely by bottom-up processing of the retinal stimuli. The present study suggests that binocular attention plays an important role in stereo capture by way of selecting and enhancing a perceptual configuration that is often ambiguous without attention involved. Stereo capture results from wallpaper's local rematching driven by binocularly attended 3-D configuration rather than retinal images.

Attention↗

Three-dimensionally slanted illusory contours capture stereopsis.

In stereo capture a stereogram of a crossed illusory figure pulls a texture bounded by the illusory contours to the same depth plane with the illusory figure. We investigated whether three-dimensionally curved and slanted illusory figures could capture a repeating background texture. According to results, stereoscopic capture was perceived when a disparate illusory contour was slanted provided that the period of the background texture was consistent with the three-dimensional geometry of the illusory surface. We suggest that stereo capture is actually induced by disparate rows of points defined by vertical cut-out sectors and the induced disparity spread is constrained by illusory contours and possible discrete matches of the background texture.

Adult↗

Posterior capsulorhexis with optic capture in pediatric cataract and intraocular lens surgery.

BACKGROUND: A major concern in pediatric intraocular lens implantation surgery is the high incidence of posterior capsule opacification. The purpose of this article is to investigate a surgical technique for maintaining a clear visual axis after pediatric cataract and intraocular lens surgery. METHODS: In a prospective evaluation, posterior capsulorhexis with optic capture as a surgical technique for preventing secondary membrane formation after pediatric cataract extraction is investigated. This technique involves capture of an intraocular lens optic through a posterior curvilinear capsulorhexis opening. Perioperative complications and incidence of secondary cataract are presented. Results will be compared to an historic cohort of patients with pediatric cataract-intraocular lens who did not have posterior continuous curvilinear capsulorhexis with optic capture. RESULTS: In 13 consecutive eyes operated on in children aged 2 to 12 years (mean, 5.8 years), there was no opacification of the visual axis at a mean follow-up of 19 months after surgery (range, 8-30 years). CONCLUSION: Posterior capsulorhexis with optic capture holds promise as a technique for preventing secondary membrane formation. This maneuver ensures centration of the posterior chamber intraocular lens while eliminating the need for an anterior vitrectomy. Apposition of the anterior and posterior capsule leaflets anterior to the optic may limit the migration of Elschnig pearls, reducing the incidence of secondary membranes and the need for additional procedures.

Cataract↗