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At least 1,531 records · Page 85Linked to original sources

PubMed2003

Analysis of complement-bound hepatitis B virus complexes by an immuno-capture polymerase chain reaction method.

Hepatitis B Virus (HBV) can be present in the circulating blood either as a free virus or as a virion-immunoglobulin (Ig) complex with or without complement. However, information regarding the complement-bound HBV circulating immune complexes (CIC) in HBV-infected patients is currently not available. In the present study, we have combined immuno-capture and polymerase chain reaction (PCR) and developed a quick method of high specificity for the detection of complement-bound HBV CIC. We found that the frequency of HBV-factor B was associated with clinical types of hepatitis B (HB) but not with that of HBV-C1q. Moreover, increased frequency at P < 0.05 were found for HBV/C1q-CIC in the group with normal total bilirubin (TBIL) and for HBV/factor B-CIC in the group with positive hepatitis B e antigen (HBeAg). These findings suggest that the immuno-capture PCR (iPCR) for the detection of HBV-bound CIC is a valuable method for analysis of the composition of the immune complexes and for the understanding of host immune response and immune pathogenesis in HBV-infected individuals. In summary, iPCR is a valuable method for analysis of the composition of the immune complexes, which may provide new and valuable insights into HBV pathogenesis.

Adolescent↗

PubMed2001

Enumeration of non-communicable disease in rural South Africa by electronic data linkage and capture-recapture techniques.

Non-communicable diseases (NCDs) are becoming increasingly common and important in developing countries, yet their enumeration is problematic. We have attempted to enumerate NCD patients in a rural district of KwazuluNatal, South Africa, using the techniques of electronic data linkage and capture-recapture (CR). We examined four major NCDs (hypertension, diabetes, asthma and epilepsy). Basic patient details were recorded onto EpiInfo software over a 6-week period, from the main hospital clinic at Hlabisa, as well as the 10 outlying peripheral health clinics. Using electronic data linkage of lists from the main hospital, the peripheral clinics, and repeat prescription cards, a district NCD register was produced of 2455 patients. The mean age was 51 +/- 16 years (1 SD) and 76% were female. Of the total NCD patients, 62% had hypertension (age 57 +/- 12 years, 82% female), 16% epilepsy (age 35 +/- 17 years, 49% female), 13% asthma (age 45 +/- 19 years, 60% female) and 12% diabetes (age 54 +/- 13 years, 61% female). Estimated population crude prevalence rates for known NCD cases on the register were 7.4% for hypertension, epilepsy 0.2%, asthma 0.2% and diabetes 0.2%. We also attempted a CR analysis to assess completeness of data, by comparing overlap between patients attending peripheral clinics, and the central Hlabisa Hospital clinic. Matching by name, age, and diagnosis proved feasible, but there was little overlap, and CR calculations were invalid because of the relative independence of sources. We conclude that NCDs are common in rural Africa, and that a simple NCD district register is a potentially feasible and inexpensive option. Capture-recapture analysis is feasible, but requires suitable lists with acceptable overlap of patients.

Adolescent↗

PubMed2003

Ultrasound for primary confirmation of mechanical capture in emergency transcutaneous pacing.

Clinical confirmation of mechanical capture in emergency transcutaneous pacing is sometimes difficult because of weak pulse volume or interference from skeletal muscle contractions. This report is of a case of using bedside ultrasound for primary confirmation of emergency transcutaneous pacemaker capture, and for differentiating between causes of persistent hypotension despite successful pacing. The technique of ultrasound-guided transcutaneous pacing is described.

Aged↗

PubMed2003

Association between esophageal squamous cell carcinoma and human papillomavirus detected by Hybrid Capture II assay.

From July 1999 to July 2000, 40 patients from the Surgical Oncology and Gastroenterology Departments of Irmandade da Santa Casa de Misericórdia de Porto Alegre were enrolled in a prospective study in order to find the frequency of human papillomavirus DNA (HPV-DNA) in esophageal squamous cell carcinoma. They were compared to 10 normal individuals (controls). Digene Inc. (Digene Corporation, Gaithersburg MD, USA) Hybrid Capture II assay, a chemoluminescent method, was used in both groups. The samples were collected from the tumor group by endoscopic biopsy in 20 cases, and by surgical specimens in the other 20 cases; the control group all underwent endoscopic biopsies. The cut-off point in the analytical assay for HPV-DNA was 1.0 pg/mL. Digene Corp. Hybrid Capture II assay tests HPV-DNA and differentiates high and low-risk for malignization virus groups. Gender, age, location of the tumor in esophagus and degree of differentiation on histology were also analyzed. There was one positive case (1.14 pg/mL), a 2.5% frequency of low-risk HPV-DNA type in the esophageal squamous cell carcinoma group (CI 95%: 0.00-7.34%). This group of patients consisted of 30 male and 10 female individuals, similar to the distribution seen in large series of such patients from the Brazilian population. The mean age was 63 years. Endoscopic samples of 10 patients who underwent endoscopic biopsy procedure for aleatory reasons, with normal looking esophageal mucosa, were tested to detect HPV-DNA, as controls. There was one positive case (1.17 pg/mL) of high-risk HPV-DNA type, resulting in a 10% positive rate. HPV-DNA is rarely found in esophageal squamous cell carcinomas: we found a 2.5% frequency, with confidence interval of 0-7.34%; and this is in the same range of controls (10% frequency, with CI: 9.1-10.9%). Both patients had a viral load quite near the 1.0 pg/mL cut-off point.

Adult↗

PubMed2000

IF-LCM: laser capture microdissection of immunofluorescently defined cells for mRNA analysis rapid communication.

BACKGROUND: The next phase of the molecular revolution will bring functional genomics down to the level of individual cells in a tissue. Laser capture microdissection (LCM) coupled with reverse transcription-polymerase chain reaction (RT-PCR) can measure gene expression in normal, cancerous, injured, or fibrotic tissue. Nevertheless, targeting of specific cells may be difficult using routine morphologic stains. Immunohistochemistry can identify cells with specific antigens; however, exposure to aqueous solutions destroys 99% of the mRNA. Consequently, there is an overwhelming need to identify specific tissue cells for LCM without mRNA loss. We report on a rapid immunofluorescent LCM (IF-LCM) procedure that allows targeted analysis of gene expression. METHODS: A LCM microscope was outfitted for epifluorescence and light level video microscopy. Heat filters were added to shield the image intensifier from the laser. Frozen sections were fluorescently labeled by a rapid one minute incubation with anti-Tamm-Horsfall antibody and an ALEXA-linked secondary antibody. Fluorescently labeled thick ascending limb (TAL) cells were detected by low light level video microscopy, captured by LCM, and mRNA was analyzed by RT-PCR for basic amino acid transporter, Tamm-Horsfall protein, and aquaporin-2. RESULTS: The immunofluorescently identified TAL could be cleanly microdissected without contamination from surrounding tubules. The recovery of RNA following rapid immunofluorescence staining was similar to that obtained following hematoxylin and eosin staining, as assessed by RT-PCR for malate dehydrogenase. CONCLUSIONS: We conclude that the new apparatus and method for the immunofluorescent labeling of tissue cells targeted for LCM can isolate pure populations of targeted cells from a sea of surrounding cells with highly acceptable preservation of mRNA. Since the TAL is minimally injured following ischemia, identification of the different responses between TAL and surrounding tissue in damaged kidneys may provide new therapeutic targets or agents for the treatment of acute renal failure.

Acridine Orange↗

PubMed2000

Analysis of segmental renal gene expression by laser capture microdissection.

UNLABELLED: Analysis of segmental renal gene expression by laser capture microdissection. BACKGROUND: The study of normal renal physiology has been greatly aided by microdissection techniques that have delineated the exceptional functional and cellular heterogeneity both along the nephron and between different nephron populations. These techniques are not widely used to study renal injury as microdissection is difficult because of tissue necrosis or fibrosis. We developed a procedure to detect specific gene expression in specific locations of the kidney in histologic sections. METHODS: The anatomic specificity of laser capture microdissection (LCM) was employed with the sensitivity of reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: LCM/RT-PCR detected mRNA for podoplanin in 2% of a single glomerulus, rat basic amino acid transporter in 6% of a single cross-section of proximal straight tubule, and renin in eight proximal convoluted tubule cross-sections. LCM/RT-PCR could isolate pure populations of proximal convoluted tubules, proximal straight tubules, and thick ascending limbs from renal histologic sections, although pure collecting ducts could not be isolated. LCM/RT-PCR localized ischemia-reperfusion-induced induction of KC/interleukin-8 primarily to the medullary thick ascending limb, and detected transforming growth factor-beta (TGF-beta) mRNA in glomeruli of a patient with membranous glomerulonephropathy. CONCLUSIONS: When used with an appropriate laser spot size, LCM/RT-PCR can measure gene expression in glomeruli or specific parts of the nephron and can study alterations in steady-state mRNA levels in animal models of renal disease. The applications, limitations, and refinements of this approach are discussed.

Animals↗

PubMed1990

Detection of drug-dependent, platelet-reactive antibodies by antigen-capture ELISA and flow cytometry.

The effectiveness of flow cytometry in the detection of drug-dependent, platelet-reactive antibodies was investigated. In studies of seven sera known to contain quinine- or quinidine-dependent, platelet-reactive antibodies, flow cytometry was 5 to 10 times more sensitive in detecting drug-dependent antibodies (DDAbs) than the 51Cr release assay, antigen-capture enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence microscopic assay. With flow cytometry, DDAbs could be detected at drug concentrations as low as 0.1 microM, or less than one-tenth the level required with other methods. Antigen-capture ELISA was not as sensitive as flow cytometry in DDAb detection, but it did allow identification of the DDAbs' target molecules. With this assay, five of the seven DDAbs recognized both the glycoprotein Ib/IX (GPIb/IX) and glycoprotein IIb/IIIa (GPIIb/IIIa) complexes, while the remaining two sera reacted only with GPIb/IX. Of 44 consecutive patients who developed thrombocytopenia while taking quinidine, DDAbs were detected by flow cytometry in 11 (25%), more than twice the number detected by other methods. In one patient who developed thrombocytopenia while taking trimethoprim/sulfamethoxazole, DDAbs could be detected only by flow cytometry. It can be concluded that flow cytometry is highly sensitive in detecting DDAbs and allows their detection at pharmacologic concentrations of the drug. Most quinidine-dependent antibodies recognize at least two different glycoprotein complexes in the platelet membrane.

Antibodies↗

PubMed2005

Termination of epicardial left ventricular tachycardia by pacing without global capture.

It is generally accepted that the diagnosis of an epicardial origin of ventricular tachycardia (VT) can be made indirectly by observing VT termination during ablation on the epicardial surface of the heart. There is a caveat, however, which is that termination of VT during radiofrequency current application on the epicardial surface could be due to extension of the lesion beyond the epicardium. Therefore, successful ablation of VT using an epicardial approach does not necessarily prove the reentrant circuit is located superficially. We present a case of a 44-year-old man with VT storm who demonstrated successful termination of VT with radiofrequency current application on the epicardial surface of the heart. This site corresponded to a site where pacing during VT resulted in termination of VT without global capture. Isolated mid-diastolic potentials were only seen at this site as well. We hypothesize that the finding of termination of VT by pacing without global capture supports the argument that the site of pacing is a critical part of the VT circuit.

Adult↗

PubMed2001

[Clinical investigations: innovative management of data. Electronic Data Capture: IOStudy Office].

In the future clinical trials will increasingly be conducted using "electronic data capture EDC" in place of documentation on paper. The utilization of computer systems and special software programs will make quality optimization possible while speeding up the documentation and data processing. In the publication the software system IOStudy Office and its modules are presented. The facilitation of the data capturing process (physician), data monitoring (monitor), and project managment are discussed in detail.

Clinical Trials as Topic↗

PubMed1995

Logarithmic correction to the probability of capture for dissipatively perturbed Hamiltonian systems.

Hamiltonian systems are analyzed with a double homoclinic orbit connecting a saddle to itself. Competing centers exist. A small dissipative perturbation causes the stable and unstable manifolds of the saddle point to break apart. The stable manifolds of the saddle point are the boundaries of the basin of attraction for the competing attractors. With small dissipation, the boundaries of the basins of attraction are known to be tightly wound and spiral-like. Small changes in the initial condition can alter the equilibrium to which the solution is attracted. Near the unperturbed homoclinic orbit, the boundary of the basin of attraction consists of a large sequence of nearly homoclinic orbits surrounded by close approaches to the saddle point. The slow passage through an unperturbed homoclinic orbit (separatrix) is determined by the change in the value of the Hamiltonian from one saddle approach to the next. The probability of capture can be asymptotically approximated using this change in the Hamiltonian. The well-known leading-order change of the Hamiltonian from one saddle approach to the next is due to the effect of the perturbation on the homoclinic orbit. A logarithmic correction to this change of the Hamiltonian is shown to be due to the effect of the perturbation on the saddle point itself. It is shown that the probability of capture can be significantly altered from the well-known leading-order probability for Hamiltonian systems with double homoclinic orbits of the twisted type, an example of which is the Hamiltonian system corresponding to primary resonance. Numerical integration of the perturbed Hamiltonian system is used to verify the accuracy of the analytic formulas for the change in the Hamiltonian from one saddle approach to the next. (c) 1995 American Institute of Physics.

Journal Article↗

PubMed2003

Folliculotropic mycosis fungoides with central nervous system involvement: demonstration of tumor clonality in intrafollicular T cells using laser capture microdissection.

BACKGROUND: Folliculotropic mycosis fungoides (MF) is a rare variant of cutaneous T-cell lymphoma, MF type, characterized by atypical lymphocytes preferentially infiltrating the hair-follicle epithelium relative to the epidermis. OBSERVATIONS: We describe a rare case of folliculotropic MF involving the central nervous system. This is also the first case in which laser capture microdissection was used to show that the atypical lymphocytes within the hair-follicle epithelium were part of the same tumor clone present in other tissue compartments. CONCLUSIONS: In reviewing the literature describing atypical lymphocytes infiltrating hair-follicle epithelium relative to the epidermis, we encourage the use of the term folliculotropic mycosis fungoides. Our case also supports previous findings that central nervous system involvement can occur in advanced MF. The successful procurement and analysis of atypical lymphocytes from hair-follicle epithelium by laser capture microscopy ushers in a new era in molecular diagnostics.

Aged↗

PubMed1987

Attention-dependent visual capture in double vision.

When the visual image of a body part, such as a finger, is doubled by a prism, the 'felt' position of that body part is captured by one of its visual images. Moving eye fixation from one to the other visual image is accompanied by a quick shift of the felt position. When focal attention is dissociated from foveation, the former determines visual capture. These new observations underline an active role of focal attention in intersensory integration and sensory-motor coordination of body parts.

Attention↗

PubMed1993

Asymmetrical effect of crossed and uncrossed disparity on stereoscopic capture.

When a real or subjective contour is superimposed on a repetitive texture or 'wallpaper pattern', the apparent depth of the elements of the wallpaper pattern may be influenced by the contour. When the contour has crossed disparity, the elements inside the contour are seen floating in the plane of the contour. This is called 'stereoscopic capture'. On the other hand, when the contour has uncrossed disparity a different, somewhat unstable, percept is seen. The wallpaper elements are either seen to form a transparent surface floating in front of the contour, or all of the elements inside and outside of the contour are seen to lie in the uncrossed plane of the inducing contour. We suggest that the asymmetry between crossed and uncrossed stereo capture is a by-product of the different roles played by crossed and uncrossed disparity in stereoscopic surface construction: specifically, crossed and, perhaps, zero disparity spreads one-directionally into a figure from a contour whereas uncrossed disparity spreads out from a figure in all directions. These points are illustrated with a series of demonstrations.

Adult↗

PubMed1994

Texture capture in Necker cubes.

Texture capture can be based on illusory contours, stereopsis, outline figures, real movement, and apparent movement. Several figures are presented that demonstrate texture capture by the use of the pictorial depth cues of the Necker cube. The textures seem affixed to the sides of the cube, follow the alternations of the cube, and extend off the picture plane. A concomitant phenomenon is a subjective rarefaction of the texture in the cube. It is suggested that boundary cues determine the assignation of texture to surfaces.

Form Perception↗

PubMed1999

Motion capture of stationary lines by apparently moving terminators.

Several studies and observations of a new form of motion capture are reported: frames containing identical rows of evenly spaced vertical lines are alternated in a standard apparent-motion paradigm. However, one vertical line in the first frame has short horizontal 'terminators' attached; the terminators are shifted to a different line in the second frame. Alternation that includes an unpatterned, nonzero interstimulus interval results in perceived motion of a vertical line along with the terminators. This motion can 'cross over' other stationary vertical lines and persists when light-filled interstimulus intervals and gaps between lines and terminators are introduced. It can also be obtained with different line sizes and spacings. The present motion capture does not appear to rely on a global-frame effect. Alternative explanations are considered.

Adult↗

PubMed2006

Transcriptome of Salmonella enterica serovar Typhi within macrophages revealed through the selective capture of transcribed sequences.

The cDNA obtained by selective capture of transcribed sequences is a complex mixture that can be used in conjunction with microarrays to determine global gene expression by a pathogen during infection. We used this method to study genes expressed by Salmonella enterica serovar Typhi, the etiological agent of typhoid fever, within human macrophages. Global expression profiles of Typhi grown in vitro and within macrophages at different time points were obtained and compared. Known virulence factors, such as the SPI-1- and SPI-2-encoded type III secretion systems, were found to be expressed as predicted during infection by Salmonella, which validated our data. Typhi inside macrophages showed increased expression of genes encoding resistance to antimicrobial peptides, used the glyoxylate bypass for fatty acid utilization, and did not induce the SOS response or the oxidative stress response. Genes coding for the flagellar apparatus, chemotaxis, and iron transport systems were down-regulated in vivo. Many cDNAs corresponding to genes with unknown functions were up-regulated inside human macrophages and will be important to consider for future studies to elucidate the intracellular lifestyle of this human-specific pathogen. Real-time quantitative PCR was consistent with the microarray results. The combined use of selective capture of transcribed sequences and microarrays is an effective way to determine the bacterial transcriptome in vivo and could be used to investigate transcriptional profiles of other bacterial pathogens without the need to recover many nanograms of bacterial mRNA from host and without increasing the multiplicity of infection beyond what is seen in nature.

Cell Line↗

PubMed2006

Tactile guidance of prey capture in Etruscan shrews.

Whereas visuomotor behaviors and visual object recognition have been studied in detail, we know relatively little about tactile object representations. We investigate a new model system for the tactile guidance of behavior, namely prey (cricket) capture by one of the smallest mammals, the Etruscan shrew, Suncus etruscus. Because of their high metabolic rate and nocturnal lifestyle, Etruscan shrews are forced to detect, overwhelm, and kill prey in large numbers in darkness. Crickets are exquisitely mechanosensitive, fast-moving prey, almost as big as the shrew itself. Shrews succeed in hunting by lateralized, precise, and fast attacks. Removal experiments demonstrate that both macrovibrissae and microvibrissae are required for prey capture, with the macrovibrissae being involved in attack targeting. Experiments with artificial prey replica show that tactile shape cues are both necessary and sufficient for evoking attacks. Prey representations are motion- and size-invariant. Shrews distinguish and memorize prey features. Corrective maneuvers and cricket shape manipulation experiments indicate that shrew behavior is guided by Gestalt-like prey descriptions. Thus, tactile object recognition in Etruscan shrews shares characteristics of human visual object recognition, but it proceeds faster and occurs in a 20,000-times-smaller brain.

Animals↗

PubMed2002

Selective immobilization of proteins to self-assembled monolayers presenting active site-directed capture ligands.

This paper describes a method for the selective and covalent immobilization of proteins to surfaces with control over the density and orientation of the protein. The strategy is based on binding of the serine esterase cutinase to a self-assembled monolayer presenting a phosphonate ligand and the subsequent displacement reaction that covalently binds the ligand to the enzyme active site. Surface plasmon resonance (SPR) spectroscopy showed that cutinase binds irreversibly to a monolayer presenting the capture ligand at a density of 1% mixed among tri(ethylene glycol) groups. The covalent immobilization is specific for cutinase, and the glycol-terminated monolayer effectively prevents unwanted nonspecific adsorption of proteins. To demonstrate that the method could be used to immobilize proteins of interest, a cutinase-calmodulin fusion protein was constructed and immobilized to the monolayer. SPR showed that calcineurin selectively associated with the immobilized calmodulin. This capture ligand immobilization method combines the advantages that the immobilization reaction is highly selective for the intended protein, the tether is covalent and, hence, stable, and the method avoids the need for synthetic modification and rigorous purification of proteins before immobilization. These characteristics make the method well suited to a range of applications and, in particular, for constructing protein microarrays.

Binding Sites↗