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Biomedical subjects

D Chien

Publications and source records attributed to D Chien.

At least 19 recordsLinked to original sources

At least five related, but distinct, hepatitis C viral genotypes exist.

Hepatitis C virus, the major causative agent of blood-borne non-A, non-B hepatitis in the world, has been the subject of considerable nucleic acid sequence analysis. Although all reported hepatitis C sequences from the United States have been represented by the prototype hepatitis C virus type 1 sequence, two groups of variant sequences have been reported in Japan. However, we have noted five distinct, but related, genotypes (I-V) throughout the world, based on detailed sequence determination and analysis of the first 1700 nucleotides and part of the nonstructural region 5 at the C terminus of the open reading frame. The nucleotide sequence for a large number of hepatitis C virus isolates spanning six continents was obtained by direct sequence analysis of PCR products after reverse transcription. Genotype was classified by using several distinct sequence motifs. We observed that most genotypes coexist in several geographic regions, including the United States, Japan, Germany, and Italy. So far, genotype V has been found only in South Africa. Interestingly, each distinct genotype seems to be maintained throughout the genome in the segments studied. These genotype distinctions should be considered when designing specific diagnostic tests, developing potential vaccines, and studying viral transmission.

Amino Acid Sequence

Improved detection of hepatitis C virus antibodies in high-risk populations.

Sera from 483 patients at high (group 1, n = 313) and lower (group 2, n = 170) risk for exposure to hepatitis C were tested for antibodies to hepatitis C using first-generation (c100-3) and second-generation enzyme-linked immunosorbent assays and four-antigen recombinant immunoblot assay. The second-generation enzyme-linked immunosorbent assay and nitrocellulose-based immunoblot assay differ from c100-3-based systems in the addition of expression products from the NS3/NS4 (c33c, c200) and putative nucleocapsid (c22-3) region of the hepatitis C genome. In group 1, the sensitivity of detection of hepatitis C antibodies was 45%, 55% and 46% by the first- and second-generation enzyme-linked immunosorbent assays and recombinant immunoblot assay, respectively. In group 2, antibodies were detected by each test system in 26%, 32% and 7% of patients, respectively. Most sera (99%) reactive with the first-generation enzyme-linked immunosorbent assay were reactive with the second-generation enzyme-linked immunosorbent assay (in group 1, 89% of these specimens demonstrated reactivity to at least one antigen with the immunoblot assay, compared with only 31% in group 2). An additional 12% (group 1) and 6% (group 2) of specimens demonstrated reactivity with the second-generation enzyme-linked immunosorbent assay only (of these, 75% [group 1] and 9% [group 2] demonstrated reactivity to at least one antigen with the immunoblot assay). Ninety-eight percent of specimens not reactive with both enzyme-linked immunosorbent assay test systems were also nonreactive by recombinant immunoblot assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Hepatocellular

Comparative evaluation of supplemental hepatitis C virus antibody test systems.

Implementation of routine blood donor screening using anti-hepatitis C virus (HCV) enzyme immunoassay (EIA) has resulted in an urgent need for well-characterized supplemental assays to confirm the presence of HCV antibodies. A comparative study of four commercially available supplemental assays is reported here: first- and second-generation versions of a strip recombinant immunoblot assay (RIBA-1 and RIBA-2), an HCV neutralization EIA, and HCV neutralization plus synthetic peptide EIA. Three hundred sixty-seven blood donor specimens that were repeatedly reactive on HCV EIA were studied. Most specimens (93%) were also evaluated by radioimmunoassay (RIA) with a six-antigen panel, and 60 selected specimens were tested for HCV RNA by the polymerase chain reaction (PCR). RIBA-1 and RIBA-2 gave concordant results with 86 percent of specimens, while an additional 13 percent were correctly classified by RIBA-2 but not RIBA-1. Neutralization EIA alone correctly identified 94 percent of the study group, while the remaining 6 percent required the peptide EIA or the combined neutralization-peptide assay system for correct classification. The RIBA-2 and neutralization-peptide assay system for correct classification. The RIBA-2 and neutralization-peptide assay systems yielded identical results for 86 percent of specimens, and these results were supported by RIA and selected PCR testing. Only 2 specimens (0.5%) were frankly discrepant, while 51 specimens were indeterminate on either (47) or both (4) assays. When either the RIBA-2 or neutralization-peptide assay yielded an indeterminate interpretation, the other system correctly classified the specimen (based on concordance with RIA and PCR data) in a high proportion (92%) of cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Viral

Improved time-of-flight MR angiography of the brain with magnetization transfer contrast.

The authors hypothesized that magnetization transfer contrast (MTC) could be used to improve flow contrast in time-of-flight (TOF) magnetic resonance (MR) angiography. Two- and three-dimensional flow-compensated gradient-echo images were obtained with and without MTC. MTC images were obtained by applying low-power radio-frequency (RF) radiation with a frequency offset from the bulk "free" water resonance frequency before the excitation RF pulse. The signal intensity of stationary tissue decreased as the power applied for the MTC pulse was increased. A smaller decrease occurred in venous signal intensity as measured in the superior sagittal sinus, and less change was seen in the arterial signal intensity as measured in the middle cerebral artery. Cerebrospinal fluid showed no MTC effect. The use of MTC improved small-vessel depiction on maximum-intensity projection images. The authors conclude that use of MTC can substantially enhance the quality of TOF MR angiography of the brain.

Adult

Fast magnetic resonance diffusion-weighted imaging of acute human stroke.

Rapid MRI of the molecular diffusion of water demonstrated cerebral infarcts in 32 patients. We studied these patients at various times following the onset of ischemic symptoms and found that diffusion-weighted imaging revealed the infarcts sooner than conventional T2-weighted spin-echo imaging did; four hyperacute infarcts were shown only by diffusion-weighted imaging. Acute infarcts had lower apparent diffusion coefficients (ADCs) than noninfarcted regions did. This relative difference in ADC reached a nadir in the first 24 hours and rose progressively thereafter. Chronic infarcts showed a relative increase in diffusion and were readily distinguishable from acute infarcts. The technique takes less than 2 minutes to apply using a standard 1.5-tesla scanner in the clinical setting. Diffusion-weighted imaging has the potential to play a role in improving the early anatomic diagnosis of stroke and therefore in the development and implementation of early stroke interventions.

Adult

Image artifacts in fast magnetic resonance imaging.

Image artifacts are unwanted, spurious signal intensities that interfere with clinical diagnosis. This article gives an overview of image artifacts in magnetic resonance imaging. We discuss the causes of these artifacts, provide clinical examples, and offer solutions to avoid them.

Artifacts

Reliable confirmation and quantitation of human immunodeficiency virus type 1 antibody using a recombinant-antigen immunoblot assay.

The recombinant DNA-derived, human immunodeficiency virus (HIV) antigen-based immunoblot assay (RIBA-HIV216) is a new supplemental (confirmatory) test developed to detect antibodies to HIV-1. The assay employs four recombinant viral antigens, corresponding to HIV-1 p24, p31, p41 and gp120 proteins, in an immunoblot format. With this assay, HIV-1 antigen reactivity was detected in all 683 infected patient serum or plasma specimens evaluated; 665 (97.6%) of these sera met the criteria for a positive interpretation, and 18 (2.6%) were classified as indeterminate. All 683 samples reacted with the recombinant gp41-equivalent protein. The first sequential enzyme immunoassay (EIA)-reactive samples collected from 33 seroconverting homosexual men reacted on RIBA-HIV216. Eleven (1.1%) of 999 EIA-negative blood donor sera reacted weakly with a single recombinant antigen (p24 or p31), whereas 13 to 48 percent had indeterminate reactions on viral lysate Western blots. One (1.5%) of 66 EIA-positive, Western blot-negative blood donor samples and 19 (29%) of 66 EIA-positive, Western blot-indeterminate blood donor samples scored indeterminate on RIBA-HIV216. Nonspecific reactivity was seen with only 1 (0.8%) of 114 patient sera containing possible interfering antibodies, whereas 33 percent of these samples had indeterminate reactions on Western blot and 35 percent had equivocal reactions on immunofluorescence assay (IFA). We conclude that the RIBA-HIV216 is approximately as sensitive as and significantly more specific than virus-derived Western blot and IFA. The RIBA-HIV216 also allows for semiquantitation of specific antibodies that may be of value in clinical staging and therapeutic monitoring.

Blotting, Western

Fast time-of-flight MR angiography with improved background suppression.

A new technique for improving contrast in time-of-flight magnetic resonance (MR) angiography is described. A selective 180 degrees radio-frequency pulse was applied before data acquisition to suppress the signal intensity of stationary tissues. Vascular images were obtained in 1 second or less by using a single-shot, rapid gradient-echo sequence in conjunction with a very short echo time to minimize flow-related dephasing. Alternatively, the data acquisition could be divided into several segments that were combined to create an image. Because of the short imaging times, abdominal single-shot images were relatively insensitive to motion. The major drawback was decreased spatial resolution and limited signal-to-noise ratio. Spatial resolution of segmented flow images was comparable with that of standard gradient-echo images, but there was a marked reduction in the signal intensity of stationary tissues. For high-resolution MR angiography, the effective background suppression obtained by means of the segmented approach can improve the quality and reliability of images created by the maximum intensity projection algorithm.

Abdomen

Fast selective black blood MR imaging.

To overcome the problems associated with gradient-echo (GRE) magnetic resonance (MR) angiography ("bright blood" imaging) and "black blood" imaging with presaturated spin-echo (SE) pulse sequences, the authors devised a new approach for black blood imaging. Their method, selective preinversion fast imaging with steady precession (turboFISP), uses a segmented GRE sequence for fast data acquisition. Nulling of vascular signal results, and stationary tissue appears bright. The method was compared with flow-compensated GRE imaging in a phantom and with GRE imaging and presaturated SE imaging in seven healthy volunteers and nine patients with various cardiac diseases. With phantoms, the selective preinversion turboFISP sequence produced better flow contrast than did GRE sequences. Selective preinversion turboFISP was often superior to SE imaging for depicting vessel lumina, particularly in patients with slowly flowing blood. Arteries appeared dark in selective black blood angiograms, but veins did not. Selective preinversion turboFISP can be used with bright blood GRE imaging to depict vessel lumina, and its capability for image acquisition within a breath hold and with cardiac gating minimizes artifacts from respiration and motion of the vessel wall.

Adult

Ultrafast imaging using gradient echoes.

Ultrafast magnetic resonance (MR) imaging techniques can reduce scan times to less than 1 s. The rapid acquisition minimizes motion artifacts that have plagued MR studies of the heart and abdomen, and facilitates dynamic studies to observe physiological function. We first discuss fast gradient-echo methods, including various spoiled and steady-state gradient-echo techniques. Ultrafast methods are then considered, with the focus on turbo-fast low-angle shot (FLASH) (also known as snapshot or subsecond FLASH) imaging. Although turbo-FLASH is a subset of gradient echo methods, there are several distinguishing features. For instance, with T1- or T2-weighted turbo-FLASH, the magnetization never reaches a steady state, so that the phase encode order becomes an important imaging parameter. Furthermore, image contrast is obtained by adjusting the magnetization preparation module, which is independent of the data acquisition module that follows. The signal behavior and strategies for contrast optimization are discussed. Potential clinical applications, including perfusion imaging, cardiac cine, breath-hold abdominal imaging, angiography, diffusion imaging, and three-dimensional studies, are explored.

Fourier Analysis

0.3-second FLASH MRI of the human heart.

Flow-suppressed FLASH MR images of the human heart have been recorded within a measuring time of 0.3 s using a 2.0-T whole-body research system (Siemens Magnetom) equipped with a conventional 10 mT m-1 gradient system. Subsecond imaging times have been achieved by reducing the repetition time to TR = 4.8 ms and by lowering the spatial resolution to 64 X 128 measured data points. The flip angle of the slice-selective radiofrequency (rf) pulses was adjusted to 10 degrees. Cardiac chambers, ventricular walls, and valves are well delineated in images from a single cardiac cycle using a field of 250 mm and a slice thickness of 8 mm. No motion artifacts were observed as a consequence of the short echo time of TE = 2.8 ms. Distinction between flowing blood and solid structures has been achieved by spatial presaturation of adjacent slices using two slice-selective 90 degrees rf pulses preceding the entire imaging sequence.

Heart

Advances in cardiac applications of subsecond flash MRI.

Flow-suppressed, subsecond FLASH MR images of the normal human heart have been obtained from single cardiac cycles using a 2.0-T whole-body MRI/MRS system (Siemens Magnetom) equipped with conventional 10 mT m-1 gradients. The present results demonstrate further technical improvements as compared to a previous report on the same subject (Magn. Reson. Med. 13:150-157; 1990). Measuring times of 139 msec and 209 msec were achieved by reducing the repetition time to TR = 4.36 msec (TE = 2.8 msec) and the spatial resolution to 32 x 128 or 48 x 128 measured data points, respectively. The flip angle was optimized to 12 degrees. Spatial pre-saturation of 60 mm thick sections adjacent to the imaging plane resulted in a suppression of the blood signal and a clear delineation of the myocardium. Oblique rotation of the imaging slice provides convenient access to the anatomical long axis and short axis views of the heart. EKG-triggered images from separate heartbeats but at different cardiac phases demonstrate that the effective time resolution is considerably less than the actual imaging time.

Electrocardiography

Localized NMR spectroscopy in vivo. Progress and problems.

Metabolites in brain and muscle of normal human volunteers have been studied by localized 1H and 31P NMR spectroscopy in vivo. Localization was achieved by means of stimulated echo (STEAM) sequences for both water-suppressed 1H NMR (TE = 20 ms) and 31P NMR (TE = 3 ms). Volumes-of-interest and measuring times selected for brain spectroscopy were 8 mL and 6.5 min for 1H NMR and 125 mL and 13 min for 31P NMR, respectively. General problems relating to spatial localization, spectral resolution, and quantitation of in vivo NMR data are discussed with respect to the nucleus and organs under investigation. They are correlated to studies of tissue extracts obtained at field strengths of 2.35 T (Bruker Biospec) and 7.0 T (Bruker MSL 300). Human studies were performed at 2.0 T on a whole-body research system (Siemens Magnetom).

Brain

Strategies to improve contrast in turboFLASH imaging: reordered phase encoding and k-space segmentation.

TurboFLASH (fast low-angle shot) sequences enable the acquisition of an image in a fraction of a second. However, unique to T1-weighted ultrafast imaging, the magnetization variation during image acquisition can produce artifacts along the phase-encoding direction. In this study, the signal behavior and nature of these artifacts were analyzed with various acquisition schemes to improve image contrast. The magnetization variation during image acquisition and its filtering effect on the image were simulated for three different approaches to T1-weighted turboFLASH imaging: standard turboFLASH with (a) monotonically ascending phase-encoding steps, (b) reordered phase encoding, and (c) k-space segmentation. Each of the modified data acquisition schemes has advantages. However, for subsecond imaging, reordered phase encoding produced improved image contrast over that of standard turboFLASH, and segmented k-space imaging gave superior tissue contrast compared with that of both standard and reordered turboFLASH, with imaging time that permits breath-hold studies.

Computer Simulation

High-speed black blood imaging of vessel stenosis in the presence of pulsatile flow.

Stenosis phantoms were created to study the ability of "black blood" methods to image a vessel stenosis in the presence of pulsatile flow. Black blood images were acquired with a modified TurboFLASH (fast low-angle shot) method that eliminates flow signal by applying a set of prepulses before segmented data acquisition. With this high-speed approach, imaging can be completed within 16 seconds. This technique was compared with conventional spin-echo black blood, gradient-echo black blood, and gradient-echo bright blood methods. Loss of flow signal, which extended beyond the site of the stenosis, was seen on the gradient-echo bright blood images. The pattern of signal loss varied with the type of stenosis. Flow voids were achieved with spin-echo black blood imaging; however, substantial ghosting artifacts were seen. With gradient-echo black blood imaging, it was difficult to eliminate all flow signal, particularly for in-plane flow. The modified TurboFLASH method produced high-quality black blood images in a fraction of the time needed for spin-echo imaging. It showed no ghosting artifacts even in the presence of pulsatile flow.

Blood Flow Velocity