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Assaying phage-borne peptides by phage capture on fibrinogen or streptavidin.

There is no simple and efficient method for assaying phage isolated from libraries without having to resort to PEG purification of the phage, or to the biotinylation or other labelling of the target molecule. We report here a method for producing 'bifunctional' phage that express two types of peptide; one peptide, fused to pVIII, will bind to immobilized fibrinogen, allowing capture of the phage out of culture supernatants; this allows the other peptide, fused to pIII or pVIII to be assayed by simple ELISA. This system has also been developed for the capture of phage bearing a streptavidin-binding peptide. The bifunctional phage are produced by bacterial cells bearing a plasmid that expresses pVIII fused either to the fibrinogen-binding peptide or to the streptavidin-binding one. Thus, when these cells are infected with a phage clone or pool to be assayed, phage will be produced whose 'capture-peptide' is produced from the plasmid and whose 'assay-peptide' is produced from the phage genome. We show here that, by this method, bifunctional phage can be produced that will bind to immobilized streptavidin or fibrinogen.

Antibody Affinity↗

Enzyme immunoassay with captured hapten. A sensitive gastrin assay with biotinyl-gastrin derivatives.

Two N-terminal biotinyl-gastrin derivatives were synthesized to investigate the effect of the length and chemical properties of the biotin-spacer on both the capture of the hapten by streptavidin or avidin adsorbed on polystyrene, and the antigenicity of the captured peptide. The observed full retainment of antibody binding capacity of the biotinyl-gastrins upon their immobilization, allowed to develop a sandwich-type ELISA with a sensitivity of one order of magnitude better than the standard ELISA with polystyrene-adsorbed gastrin. This hapten capture system reduces desorption particularly pronounced for low mass peptides, and avoids possible modifications or suppression of epitopes by the adsorption process with concomitant reduction of antibody binding affinity of the antigen. This new type of assay procedure may also represent a useful tool particularly for epitope mapping with relatively low mass synthetic protein fragments.

Amino Acid Sequence↗

Homosynaptic and heterosynaptic inhibition of synaptic tagging and capture of long-term potentiation by previous synaptic activity.

Long-term potentiation (LTP) is an enhancement of synaptic strength that may contribute to information storage in the mammalian brain. LTP expression can be regulated by previous synaptic activity, a process known as "metaplasticity." Cell-wide occurrence of metaplasticity may regulate synaptic strength. However, few reports have demonstrated metaplasticity at synapses that are silent during activity at converging synaptic inputs. We describe a novel form of cell-wide metaplasticity in hippocampal area CA1. Low-frequency stimulation (LFS) decreased the stability of long-lasting LTP ["late" LTP (L-LTP)] induced later at the same inputs (homosynaptic inhibition) and at other inputs converging on the same postsynaptic cells (heterosynaptic inhibition). Significantly, heterosynaptic inhibition of L-LTP also occurred across basal and apical dendrites ("heterodendritic" inhibition). Because transient early LTP (E-LTP) was not affected by previous LFS, we examined the effects of LFS on the consolidation of E-LTP to L-LTP. The duration of E-LTP induced at one set of inputs can be extended by capturing L-LTP-associated gene products generated by previous activity at other inputs to the same postsynaptic neurons. LFS applied homosynaptically or heterosynaptically before L-LTP induction did not impair synaptic capture by subsequent E-LTP stimulation, suggesting that LFS does not impair L-LTP-associated transcription. In contrast, LFS applied just before E-LTP (homosynaptically or heterosynaptically) prevented synaptic tagging, and capture of L-LTP expression. Thus, LFS inhibits synaptic tagging to impair expression of subsequent L-LTP. Such anterograde inhibition represents a novel way in which synaptic activity can regulate the expression of future long-lasting synaptic plasticity in a cell-wide manner.

Animals↗

[Capture and recapture method: a new methodology for epidemiological research].

Although capture-recapture method is not new, their use in epidemiological studies has so far been limited. The method was developed by animal ecologists to estimate the size of various species populations, but in recent years it has been used to measure the incidence and prevalence of a variety of non-communicable diseases. Capture-recapture allows more accurate estimates to be made than is the case with traditional methods and is more cost-effective. The method is ideally suited to studying rare or elusive populations, such as intravenous drug misusers; but they are highly adaptable and have been used to study populations as diverse as street working prostitutes and the human red-cell volume. The objectives of this review are to explain the mathematical theory behind capture-recapture, give examples of its use and application in clinical epidemiology and out line some of the modifications that have been made to the theory, which take into account the complex nature of samples used in clinical research. An increasing number of medical epidemiologists have started to use the method, and its potential for future epidemiological research is enormous.

Epidemiologic Methods↗

The epidemiologic importance of Triatoma brasiliensis as a chagas disease vector in Brazil: a revision of domiciliary captures during 1993-1999.

To clarify the epidemiologic importance of Triatoma brasiliensis, the most important Chagas disease vector in the Northeastern of Brazil, capture data related to this species, its distribution, capture index, and percentages of natural infection by Trypanosoma cruzi were examined in 12 different Brazilian states. The Brazilian National Health Foundation collected these data from 1993 to 1999, a period during which a total of 1,591,280 triatomines (21 species) were captured in domiciles within the geographic range of T. brasiliensis. Of this total, 422,965 (26.6%) were T. brasiliensis, 99.8% of which were collected in six states, and 54% in only one state (Ceará). The percentage of bugs infected with T. cruzi varied significantly among states, ranging from 0% (Goiás, Maranhão, Sergipe, and Tocantins) to more than 3% (Alagoas, Minas Gerais, and Rio Grande do Norte) with an average of 1.3%. This latter value represents a dramatic reduction in the natural infection percentages since 1983 (6.7%) suggesting that, despite the impossibility of eradicating this native species, the control measures have significantly reduced the risk of transmission. However, the wide geographic distribution of T. brasiliensis, its high incidence observed in some states, and its variable percentages of natural infection by T. cruzi indicate the need for sustained entomological surveillance and continuous control measures against this vector.

Animals↗

[Capture-recapture methodology: an option for surveillance of non-communicable diseases in the elderly].

Capture-recapture methodology is used in Ecology to estimate the total size of wild animal populations. This method can be used in Epidemiology to estimate the incidence and prevalence of communicable and non-communicable diseases in a relatively inexpensive and quick way. Surveillance systems based on the use of this methodology are a potential alternative for monitoring non-communicable diseases in Brazil. This paper presents a brief review of fundamental capture-recapture methodology and its applications to Epidemiology. We also present a theoretical model for implementation of a diabetes mellitus surveillance system in the elderly using publicly available morbidity and mortality data sources and the capture-recapture methodology.

Aged↗

[Performance of cervical cytology with review by different observers and hybrid capture II in the diagnosis of cervical intraepithelial neoplasia grades 2 and 3].

UNLABELLED: To evaluate the performance of initial cervical cytology and that collected at the reference service with a review conducted by different observers and techniques, as well as hybrid capture II, in the diagnosis of high-grade cervical intraepithelial neoplasia (CIN), 105 women attended from August 2000 to June 2001 for preneoplastic atypia upon cervical cytology were included. A new cervical cytology and hybrid capture II for DNA-HPV were conducted in all the patients. Cervical biopsies were taken in 91 women. Performance of the investigative procedures was described by estimating the sensitivity, specificity, and positive likelihood ratio (PLR), with a 95% confidence interval (95% CI), considering histological diagnosis the gold standard. RESULTS: initial cervical cytology showed sensitivity and specificity of 57% and 82% for diagnosis of CIN grades 2 and 3, with a PLR of 3.2 (95% CI: 1.5-6.8). Referral cervical cytology showed a sensitivity and specificity for CIN 2 and 3 of 79% and 84%, respectively, with a PLR of 5.0 (95% CI: 2.5-10.0). Sensitivity (86%), specificity (80%), and PLR (4.3) were similar when a second observer (using a routine technique) reviewed the slide. Using rapid revision by a third observer, the sensitivity was significantly lower (64%). Hybrid capture II showed a high sensitivity (100%), low specificity (43%), and low PLR (1.7, 95% CI: 1.4-2.2).

Colposcopy↗

[Enterobacteria isolated from cockroaches (Periplaneta americana) captured in a Brazilian hospital].

OBJECTIVE: To isolate and identify microorganisms from cockroaches that were captured in a public hospital and to test the antimicrobial susceptibility of these microorganisms. METHODS: Cockroaches were captured in the morning and at night. They were placed in flasks rinsed with 70% alcohol, transferred to sterilized flasks, and then taken to the laboratory. Only cockroaches captured whole and live were utilized for the study. After being immobilized at 0 degree C, each cockroach was placed in a test tube with sterile saline solution (0.8%) and then homogenized. The resulting solution was then placed in the following five culture media: MacConkey agar, nutrient broth, brain-heart infusion agar, Sabouraud agar, and mannitol. The cultures were examined using a stereomicroscope, and colony-forming units were counted. The disk diffusion test was used to determine antimicrobial susceptibility. RESULTS: We found a 56% prevalence of enterobacteria and an 18% prevalence of coagulase-negative staphylococci. Fifteen species of enterobacteria were identified. The most frequent were Klebsiella pneumoniae (17%), Enterobacter aerogenes (14%), Serratia marcescens (13%), Hafnia alvei (12%), Enterobacter gergoviae and Enterobacter cloacae (each 9%), and Serratia spp. (6%). Both the enterobacteria and the coagulase-negative staphylococci showed significant resistance to antimicrobials, including oxacillin. CONCLUSIONS: The prevalence of enteropathogenic bacteria and coagulase-negative staphylococci isolated from Periplaneta americana cockroaches in the studied hospital reflects the weakness of the measures adopted both for vector control and for antimicrobial use. The results show the need to implement effective health-institution programs focusing on hygiene and the rational use of antimicrobials.

Animals↗

Mesh size and bird capture rates in Mato Grosso do Sul State, Brazil.

Mist-nets alternating 36-mm and 61-mm mesh in woods and low vegetation of "cerrado" (Brazilian savanna) tested bird-capture efficiency relative to bird length and mass. Of 1,296 birds captured and 102 species, 785 (93 species) were with 36-m mesh and 511 (69 species) with 61-mm mesh. The 61-mm mesh improved capture rates only for some larger species; so, in general, 36-mm mesh mist-nets are more appropriate for field work in "cerrado" areas.

Animals↗

Effects of routine late-season field operations on numbers of boll weevils (Coleoptera: Curculionidae) captured in large-capacity pheromone traps.

Flat and cylindrical adhesive boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), pheromone traps captured significantly more (P < or = 0.05) boll weevils than the Hercon (Hercon Environmental, Emigsville, PA) trap during the late cotton-growing season, and larger adhesive areas were associated with higher captures; a flat plywood board collected the most boll weevils because it had the largest surface area. The flat board trap, chosen for measuring large late-season adult boll weevil populations common to the Lower Rio Grande Valley of Texas in 2000 and 2001, collected more (P < or = 0.05) weevils when deployed in proximity to natural and cultivated perennial vegetation, and mean numbers of captured boll weevils were higher (P < or = 0.0001) on the leeward sides of the board traps than on the windward sides. The board trap had an estimated potential capacity of approximately 27,800 boll weevils, and the large capacity of the board trap allowed for more accurate measurements of large adult boll weevil populations than the more limited Hercon trap. Measurement of adult boll weevil numbers after the routine field operations of defoliation, harvest, shredding, and stalk-pulling, demonstrated that large populations of boll weevils persist in cotton fields even after the cotton crop has been destroyed. Increases (P < or = 0.05) in the percentage variation of trapped boll weevils relative to the numbers collected just before each field operation were observed after defoliation, harvest, shredding, and stalk-pulling, but the percentage variations followed a quadratic pattern with significant correlation (P < 0.0001; 0.59 < adjusted r2 < 0.73). Numbers of adult boll weevils caught on board traps deployed at 15.24-m intervals on windward and leeward edges of cotton fields suggested that boll weevil populations in flight after field disturbances might be affected by large-capacity trapping.

Agriculture↗

Trap height and orientation of yellow sticky traps affect capture of Chaetocnema pulicaria (Coleoptera: Chrysomelidae).

Field studies were conducted in Iowa during 2001 and 2002 to determine the optimal sampling height and orientation for using yellow sticky cards to monitor populations of Chaetocnema pulicaria Melsheimer, the vector of the bacterial pathogen Pantoea stewartii subsp, stewartii, the causal organism of Stewart's disease of corn, Zea mays L.. Sticky cards were placed at five different heights (0.15, 0.30, 0.45, 0.60, and 0.90 m) and three orientations (horizontal, vertical, and 30 degree angle) at three locations (Ames, Crawfordsville, and Sutherland) in 2001 and two locations (Crawfordsville and Johnston) in 2002. No statistical differences were observed among the placement combinations for individual sampling periods or for the total number of C. pulicaria captured in 2001. In 2002, the 0.30 m and vertical cards captured significantly (1.1-35 times) more C. pulicaria than any other placement combination during sampling throughout August at both Crawfordsville and Johnston. Also, the cumulative number of C. pulicaria captured by the 0.30 m and vertical cards was significantly higher than all other placement combinations. This information is important in the development of sampling protocols to aid growers in making management decisions. These management decisions include where and when to apply foliar insecticides during the corn growing season to control C. pulicaria populations, thereby reducing the risk for Stewart's disease of corn.

Animals↗

Broadcasts of wing-fanning vibrations recorded from calling male Ceratitis capitata (Diptera: Tephritidae) increase captures of females in traps.

Female Mediterranean fruit flies, Ceratitis capitata (Wiedemann), from the sterile-male rearing facility in El Pino, Guatemala, were exposed to broadcasts of wing-fanning vibrations recorded from males engaged in calling behavior to investigate the feasibility of developing a female-selective acoustic trap. The recorded signals had frequent amplitude fluctuations and peak frequencies approximately 350 Hz, typical of signals observed in previous studies of Mediterranean fruit fly acoustic behavior. Females did not exhibit long-distance phonotaxis, but remained near a speaker significantly longer when the sounds were broadcast at 103-107 dB than when the speaker was silent. In addition, significantly higher percentages of females were captured by yellow adhesive traps next to a broadcasting speaker than by traps next to a silent mimic. Additional bioassays were conducted with synthetic, 350-Hz tones produced by a thermoacoustic tube as well as with silent mimics of the different sound sources to examine the relative responsiveness of female Mediterranean fruit flies to traps with different acoustic and visual features. The visual attributes of the different sound source assemblies significantly affected capture rates. The range over which the broadcast significantly increased the percentage of female captures was <0.5 m, which may limit the utility of these acoustic cues in large-scale trapping programs. However, the findings of this study do justify further testing of whether optimized short-range acoustic signals could be used to augment longer range pheromonal and visual cues to improve the efficacy of female-selective traps.

Acoustics↗

Physiologic manifestations of stress from capture and restraint of free-ranging male African green monkeys (Cercopithecus aethiops).

Adrenal gland weights, stomach mucosal lesions, and morning serum cortisol and prolactin levels were measured in 15 juvenile and adult male African green monkeys (Cercopithecus aethiops) that were shot by a hunter, euthanized after 24 hr of captivity, or euthanized after 45 days of captivity and intermittent blood sampling. Hormone levels were measured in seven additional males that had been in captivity for 7 mo. Mean serum cortisol concentrations were significantly lower in free-ranging wild monkeys at the time they were shot than in the monkeys after 1 day in captivity. Cortisol concentrations were significantly higher in wild-caught monkeys on the day after capture than they were in the same animals after 18 and 26 days of captivity. Cortisol concentrations were also significantly higher in the wild-caught monkeys 18 days after capture than in the laboratory-habituated monkeys in captivity for 7 mo. Mean prolactin concentration was significantly lower in the wild-caught monkeys on day 2 after capture, and the levels increased gradually to 45 days in captivity and was highest in monkeys that had been captive for 7 mo.

Adrenal Glands↗

Development of an antibody-based capture enzyme-linked immunosorbent assay for detecting Echinostoma caproni (Trematoda) in experimentally infected rats: kinetics of coproantigen excretion.

The present study reports on the development of a coproantigen capture enzyme-linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory-secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49-56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections.

Animals↗

Electron-capturing derivatization of neutral steroids for increasing sensitivity in liquid chromatography/negative atmospheric pressure chemical ionization-mass spectrometry.

Derivatization of neutral steroids for increasing sensitivity in liquid chromatography/negative atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) has been examined. Under APCI conditions, gas-phase electrons are provided by the corona discharge and captured by electron-affinitive compounds. In negative APCI-MS, therefore, ultrahigh sensitivity can be obtained by tagging neutral steroids, whose ionization efficiencies are low in the conventional APCI-MS, with electron-capturing moieties, such as a nitro group. We synthesized various boronic acid and hydrazine derivatives having electron-capturing moieties as derivatization reagents for 1,2-diol compounds and oxosteroids, respectively. Among reagents examined, those having the 2-nitro-4-trifluoromethylphenyl moiety were most effective in increasing sensitivity. That is, the detection responses of the derivatives with these reagents were increased by several to more than 200-fold over intact steroids, where limits of detection were some picograms. The developed derivatization procedures were applied to analyses of small amounts of steroids in human plasma and gave satisfactory results.

Boronic Acids↗

A simple method of capturing PACS and other radiographic images for digital teaching files or other image repositories.

OBJECTIVE: We sought to develop an easy-to-use method of capturing and storing radiographic images. CONCLUSION: The method that we developed can capture any digital image-including an image from a picture archiving and communication system (PACS)-using widely available, inexpensive software. Our method is easy to learn, simple to use, and inexpensive to implement. It is adaptable in a wide range of networking environments and can capture and store images rapidly for a variety of uses. It can be used without interfering with clinical workflow at the PACS workstation.

Humans↗

Capture-recapture method in the epidemiology of type 2 diabetes: a contribution from the Verona Diabetes Study.

OBJECTIVE: The present investigation used data from the Verona Diabetes Study to verify a main assumption of the capture-recapture method (source independence) and to characterize the subgroup of known diabetic patients missed by all sources whose number is estimated by the capture-recapture method. RESEARCH DESIGN AND METHODS: The Verona Diabetes Study identified 7,148 type 2 diabetic patients on 31 December 1986 using 3 different sources: family physicians, a diabetes center, and a drug prescription database. Completeness of ascertainment was estimated with traditional methods based on the hypergeometric distribution and with a log-linear model. RESULTS: Identification sources were not independent because the drug prescription database was positively related to family physicians and negatively related to the diabetes center (P < 0.001). Thus, completeness of ascertainment was overestimated (87.5% [95% CI 86.3-88.8]) when using only family physicians and the drug prescription database and underestimated (45.9% [43.9-48.1]) when using only the diabetes center and the drug prescription database. Because of characteristics contributing to variable "catchability" (probability of ascertainment), the estimated proportion of ascertainment increased with increasing time since diagnosis from 65.6% in the first tertile (<6 years) to 91.5% in the third tertile (>12 years); moreover, the ascertainment was estimated to be nearly complete (97.9%) for insulin-treated patients and scanty (28.9%) for diet-treated patients. CONCLUSIONS: Because identification sources are likely to be dependent, the capture-recapture method should be used with caution in diabetes epidemiology and possibly when at least 3 sources are available. The subgroup of diabetic patients whose existence is inferred by this technique likely consists of newly diagnosed patients with mild disease severity.

Aged↗

Prevalence of Salmonella spp in selected birds captured on California dairies.

OBJECTIVE: To determine the prevalence of Salmonella spp in wild birds commonly found on California dairies. DESIGN: Prospective study. ANIMALS: 7 selected species of birds were captured on 9 dairies in Kings and Tulare counties, California. PROCEDURE: Birds were captured (using traps and nets) and euthanatized, and the entire gastrointestinal tract was removed. Contents of the gastrointestinal tract were subjected to culture for Salmonella spp. RESULTS: 892 birds were captured, and Salmonella spp were isolated from 22 birds. The prevalence by dairy ranged from 0.7 to 16.7%, whereas the prevalence by bird species ranged from 1.2 to 3.2%. Cowbirds and English sparrows had the highest prevalence of Salmonella organisms. Five serotypes of Salmonella organisms were isolated, including Meleagridis, Montevideo, Muenster, Typhimurium, and an untyped serotype. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of our findings, birds that commonly inhabit California dairies harbor Salmonella organisms. However, because of the low prevalence of Salmonella organisms in birds and the Salmonella serotypes isolated, birds are not important reservoirs of Salmonella organisms on California dairies.

Animals↗