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J Hsieh

Publications and source records attributed to J Hsieh.

At least 19 recordsLinked to original sources
PubMed2004

A novel reconstruction algorithm to extend the CT scan field-of-view.

For various reasons, a projection dataset acquired on a computed tomography (CT) scanner can be truncated. That is, a portion of the scanned object is positioned outside the scan field-of-view (SFOV) and the line integrals corresponding to those regions are not measured. A projection truncation problem causes imaging artifacts that lead to suboptimal image quality. In this paper, we propose a reconstruction algorithm that enables an adequate estimation of the projection outside the SFOV. We make use of the fact that the total attenuation of each ideal projection in a parallel sampling geometry remains constant over views. We use the magnitudes and slopes of the projection samples at the location of truncation to estimate water cylinders that can best fit to the projection data outside the SFOV. To improve the robustness of the algorithm, continuity constraints are placed on the fitting parameters. Extensive phantom and patient experiments were conducted to test the robustness and accuracy of the proposed algorithm.

Algorithms↗
PubMed2001

E. coli Rep oligomers are required to initiate DNA unwinding in vitro.

E. coli Rep protein is a 3' to 5' SF1 superfamily DNA helicase which is monomeric in the absence of DNA, but can dimerize upon binding either single-stranded or duplex DNA. A variety of biochemical studies have led to proposals that Rep dimerization is important for its helicase activity; however, recent structural studies of Bacillus stearothermophilus PcrA have led to suggestions that SF1 helicases, such as E. coli Rep and E. coli UvrD, function as monomeric helicases. We have examined the question of whether Rep oligomerization is important for its DNA helicase activity using pre-steady state stopped-flow and chemical quenched-flow kinetic studies of Rep-catalyzed DNA unwinding. The results from four independent experiments demonstrate that Rep oligomerization is required for initiation of DNA helicase activity in vitro. No DNA unwinding is observed when only a Rep monomer is bound to the DNA substrate, even when fluorescent DNA substrates are used that can detect partial unwinding of the first few base-pairs at the ss-ds-DNA junction. In fact, under these conditions, ATP hydrolysis causes dissociation of the Rep monomer from the DNA, rather than DNA unwinding. These studies demonstrate that wild-type Rep monomers are unable to initiate DNA unwinding in vitro, and that oligomerization is required.

Adenosine Triphosphate↗
PubMed2001

High-throughput, semi-automated determination of a cyclooxygenase II inhibitor in human plasma and urine using solid-phase extraction in the 96-well format and high-performance liquid chromatography with post-column photochemical derivatization-fluorescence detection.

Compound I, 5-chloro-3-(4-methanesulfonylphenyl)-6'-methyl-[2,3']bipyridinyl, has been found to be a specific inhibitor of the enzyme cyclooxygenase II (COX II). The anti-inflammatory properties of this compound are currently being investigated. HPLC assays for the determination of this analyte in human plasma and human urine have been developed. Isolation of I and the internal standard (II) was achieved by solid-phase extraction (SPE) in the 96-well format. A C8 SPE plate was used for the extraction of the drug from human plasma (recovery >90%) while a mixed-mode (C8/Cation) SPE plate was used to isolate the analytes from human urine (recovery approximately 71%). The analyte and internal standard were chromatographed on a Keystone Scientific Prism-RP guard column (20 x 4.6 mm) connected to a Prism-RP analytical column (150 x 4.6 mm), using a mobile phase consisting of 45% acetonitrile in 10 mM acetate buffer (pH = 4); the analytes eluted at retention times of 5.2 and 6.9 min for I and II, respectively. Compounds I and II were found to form highly fluorescent products after exposure to UV light (254 nm). Thus, the analytes were detected by fluorescence (lambda(ex) = 260 nm, lambda(em) =375 nm) following post-column photochemical derivatization. Eight point calibration curves over the concentration range of 5-500 ng/ml for human plasma and human urine yielded a linear response (R2>0.99) when a 1/y weighted linear regression model was employed. Based on the replicate analyses (n = 5) of spiked standards, the within-day precision for both assays was better than 7% C.V. at all points on the calibration curve; within-day accuracy was within 5% of nominal at all standard concentrations. The between-run precision and accuracy of the assays, as calculated from the results of the analysis of quality control samples, was better than 8% C.V. and within 8% of nominal. I was found to be stable in human plasma and urine for at least 8 and 2 months, respectively. In addition, the human plasma assay was semi-automated in order to improve sample throughput by utilizing a Packard liquid handling system and a Tom-Tec Quadra 96 SPE system. The precision and accuracy of the semi-automated procedure were comparable to the manual procedure. Over 5000 clinical samples have been analyzed successfully using these methods.

Automation↗
PubMed2001

Determination of a beta(3)-agonist in human plasma by LC/MS/MS with semi-automated 48-well diatomaceous earth plate.

Methods for the determination of a beta(3)-agonist (A) in human plasma were developed and compared based on high-performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection using a turbo ion spray (TIS) interface. Drug and internal standard were isolated from plasma by three sample preparation methods, liquid-liquid extraction, Chem Elut cartridges and 48-well diatomaceous earth plates, that successively improved sample throughput for LC/MS/MS. MS/MS detection was performed on a PE Sciex API 365 tandem mass spectrometer operated in positive ion mode and using multiple reaction monitoring (MRM). The precursor/product ion combinations of m/z 625/607 and 653/515 were used to quantify A and internal standard, respectively, after chromatographic separation of the analytes. Using liquid-liquid extraction and Chem Elut cartridges, the assay concentration range was 0.5-100 ng/ml. Using diatomaceous earth plates, the concentration range of the assay was extended to 0.5-200 ng/ml. For all three assays, the statistics for precision and accuracy is comparable. The assay accuracy ranged from 91-107% and intraday precision as measured by the coefficient of variation (CV) ranged 2-10%. The sample throughput was tripled when the diatomaceous earth plate method was compared with the original liquid-liquid extraction method.

Adrenergic beta-3 Receptor Agonists↗
PubMed2001

Investigation of the slice sensitivity profile for step-and-shoot mode multi-slice computed tomography.

Multislice compound tomography (MCT) is one of the recent technology advancements in CT. Compared to single slice CT, MCT significantly improves examination time, x-ray tube efficiency, and contrast material utilization. Although the scan mode of MCT is predominately helical, step-and-shoot (axial) scans continue to be an important part of routine clinical protocols. In this paper, we present a detailed investigation on the slice sensitivity profile (SSP) of MCT in the step-and-shoot mode. Our investigation shows that, unlike single slice CT, the SSP for MCT exhibits multiple peaks and valleys resulting from intercell gaps between detector rows. To fully understand the characteristics of the SSP, we developed an analytical model to predict the behavior of MCT. We propose a simple experimental technique that can quickly and accurately measure SSP. The impact of the SSP on image artifacts and low contrast detectability is also investigated.

Image Processing, Computer-Assisted↗
PubMed2001

[Comparison of the effects of two different breastcare methods].

The purpose of this study was to compare the effects of two breastcare methods-one with moist heating pad (the pad group) and the other with moist hot towel (the towel group). Data were collected from May, 1996 to April, 1997 in a southern hospital in Taiwan. Mothers who breastfed, had no postpartum complications and had a healthy newborn were recruited in the study. Mothers who breastfed less than 2 days and those newborns without a complete body weight record were excluded. The numbers of participants were 94 in the towel group and 52 in the pad group. The tools used in this study were the questionnaire developed by the researchers, the heating pad, and the newborn body weight scale. The results revealed that: (1) there is no differences in the changes of newborns' body weight between the two groups; (2) the mothers in the pad group felt more comfortable, convenient and private than those in the towel group; (3) nurses spent less time helping mothers in the pad group than those in the towel group. The conclusion was that the heating pad could substitute for the hot towel in the breastcare method.

Adolescent↗
PubMed2000

High-throughput simultaneous determination of the HIV protease inhibitors indinavir and L-756423 in human plasma using semi-automated 96-well solid phase extraction and LC-MS/MS.

A method for the simultaneous determination of the HIV protease inhibitors indinavir and L-756423, in human plasma has been developed. Plasma samples (0.5 ml) were extracted using a 3M Empore 96-well plate in the mixed phase cation exchange (MPC) format. The extraction method was automated through the application of both the Packard 204DT and TOMTEC Quadra 96 work stations, and the resulting extracts were analyzed using a PE-Sciex API-3000 LC-MS/MS with a heated nebulizer interface (500 degrees C). The assay was linear in the concentration range 1-2500 ng/ml for indinavir and 5 2500 ng/ml for L-756423 when 0.5-ml aliquots of plasma were extracted. Recoveries of indinavir and L-756423 were greater than 76 and 80%, respectively, over the calibration curve range when using the described sample preparation method. Within-batch precision and accuracy for the quantitation of indinavir over the range 1-2500 ng/ml were 5.4% R.S.D. or less and within 4.0%, respectively. Within-batch precision and accuracy for the quantitation of L-756423 over the range 5-2500 ng/ml were 5.3% R.S.D. or less and within 3.4%, respectively. Interbatch variability for the analysis of indinavir QC samples at low (3 ng/ml), middle (250 ng/ml) and high (2250 ng/ml) were 3.2, 2.9, and 1.9%, respectively. Interbatch variability for the analysis of L-756423 QC samples at low (15 ng/ml), middle (250 ng/ml) and high (2250 ng/ml) concentration were 2.0, 2.5, and 3.3%, respectively. The validated assay was used in support of human clinical trials.

Anti-HIV Agents↗
PubMed2000

Sparing of neuronal function postseizure with gene therapy.

Numerous studies have demonstrated that gene therapy interventions can protect neurons from death after neurological insults. In nearly all such studies, however, "protection" consists of reduced neurotoxicity, with no demonstrated preservation of neuronal function. We used a herpes simplex virus-1 system to overexpress either the Glut-1 glucose transporter (GT) (to buffer energetics), or the apoptosis inhibitor Bcl-2. Both decreased hippocampal neuron loss to similar extents during excitotoxic insults in vitro and in vivo. However, the mediating mechanisms and consequences of the two interventions differed. GT overexpression attenuated early, energy-dependent facets of cell death, blocking oxygen radical accumulation. Bcl-2 expression, in contrast, blocked components of death downstream from the energetic and oxidative facets. Most importantly, GT- but not Bcl-2-mediated protection preserved hippocampal function as assessed spatial maze performance. Thus, gene therapeutic sparing of neurons from insult-induced death does not necessarily translate into sparing of function.

Animals↗
PubMed2000

Using technetium-99M dimercaptosuccinic acid renal cortex scintigraphy to differentiate acute pyelonephritis from other causes of fever in patients with spinal cord injury.

OBJECTIVES: To differentiate acute pyelonephritis (APN) from fever due to other sources in patients with spinal cord injury by using technetium-99m dimercaptosuccinic acid (DMSA) renal cortex scintigraphy (DMSA scan). METHODS: A total of 24 patients with spinal cord injury were admitted with fever. DMSA scans were performed on all patients. The final determination of the cause of the fever was based on the medical history, physical examination, laboratory evaluation, and imaging studies. RESULTS: DMSA scan accurately diagnosed APN in 12 patients with inflammation on the DMSA scan. In 5 patients with scarred lesions on the DMSA scans and in 7 patients with negative DMSA scan, the fever was attributed to other causes. The sensitivity and specificity of the DMSA scan for detecting APN were both 100%. CONCLUSIONS: The DMSA scan is a valuable adjunct in the evaluation of fever in patients with SCI.

Acute Disease↗
PubMed2000

Ofloxacin for the treatment of urinary tract infections and biofilms in spinal cord injury.

Forty two paraplegic and quadriplegic hospitalized spinal cord injured patients with urinary tract infections (UTI) were included in a double blind, randomized treatment study comparing 7 days ofloxacin (300 mg bd) with trimethoprim-sulphamethoxazole (TMPSMX; 160-800 mg bd) or an alternative, chosen because of resistance to TMPSMX. The 4-day clinical cure rate, defined as an asymptomatic patient with sterile urine, was 90% (19/21) with ofloxacin, significantly greater than 48% (10/21) for the comparison group (P=0.003) and the rate at end of therapy was 90% (19/21) with ofloxacin, against 57% (12/21) (P=0.015). Bacterial biofilms were detected on bladder epithelial cells in 39/41 (95%) patients. The biofilm score fell significantly following ofloxacin therapy (P < 0.001) or alternative therapy (P < 0.001). Ofloxacin treatment led to significantly more biofilm eradication than the other antibiotic group on day 4 (62 vs. 24%) (P=0.005) and day 7 (67 vs. 35%) (P=0.014). The study showed that ofloxacin was better than TMPSMX and alternatives at relieving clinical infection and eradicating bladder cell biofilms.

Administration, Oral↗
PubMed2000

Tomographic reconstruction for tilted helical multislice CT.

One of the most recent technical advancements in computed tomography (CT) is the introduction of multislice CT (MCT). Because multiple detector rows are used for data acquisition, MCT offers higher volume coverage, faster scan speed, and reduced X-ray tube loading. Recognizing its unique data-sampling pattern, several image reconstruction algorithms were developed. These algorithms have been shown to be adequate in producing clinically acceptable images. Recent studies, however, have revealed that the image quality of MCT can be significantly degraded when helical data are acquired with a tilted gantry. The degraded image quality has rendered this feature unacceptable for clinical usage. In this paper, we first present a detailed investigation on the cause of the image quality degradation. An analytical model is derived to provide a mathematical basis for correction. Several compensation schemes are subsequently presented, and a detailed performance comparison is provided in terms of spatial resolution, noise, computation efficiency, and image artifacts.

Head↗
PubMed2000

Investigation of a solid-state detector for advanced computed tomography.

Utilization of solid-state detectors for computed tomography (CT) has been the focus of many studies. Previous phantom and clinical experiments have shown that one of the important performance parameters for the solid-state detector is the primary speed and afterglow. In this paper, we present a detailed investigation on the signal decay characteristics of the HiLight (GE Medical Systems, Milwaukee, WI) scintillating detector. The detector primary speed and afterglow are modeled by a multiexponential function and fully characterized by a set of time constants and relative strengths. The sensitivity of these parameters to X-ray photon energy, detector aging, and radiation exposure is then established and analyzed. No statistically significant variation is observed in these parameters due to changes in the above external variables. The impact of various decay time constants on CT image quality, such as spatial resolution, noise, and artifacts, is subsequently illustrated with computer simulations and phantom experiments. Finally, an algorithmic correction scheme is derived to compensate for detector afterglow. The correction scheme employs a recursive filter to remove adverse effects of the detector decay on image quality. Experimental results have shown that the correction scheme successfully restores system spatial resolution, produces a more homogeneous noise pattern, and eliminates ring-band image artifacts due to detector afterglow. The effectiveness and robustness of the correction scheme are demonstrated by extensive phantom and clinical experiments.

Artifacts↗
PubMed2000

Tumor stroma as the main source of inhibin production in ovarian epithelial tumors.

PROBLEM: Elevated serum inhibin levels have been found in ovarian cancer patients; however, the source of the elevated inhibin is uncertain. Previous studies of activin in human ovarian cancer suggest that activin may promote the growth of the tumor. The aims of this study were to examine the source of elevated inhibin from ovarian epithelial tumors (OETs) and to preliminarily investigate the role of the gonadotropin-inhibin/activin relationship in the development of OET. METHOD OF STUDY: The protein and mRNA expression of alpha and betaA subunits of inhibin/activin were examined by immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR) in 120 OETs, including 30 benign cystadenomas, 30 borderline tumors, and 60 carcinomas. Stromal and epithelial cells were microdissected from 23 OETs to further examine the expression of alpha and betaA subunits by RT-PCR. Dimeric inhibin A and activin A production were measured by using the two-site ELISA from three OET cell lines in culture under treatment of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). RESULTS: betaA subunit was expressed in the epithelial component of 100% of the cystadenomas, in 80% of borderline tumors, and in 75% of the carcinomas, but not in tumor stroma. Inhibin alpha expression was not found in the epithelium of all OETs studied, but focal inhibin alpha immunoreactivity was seen in the tumor stroma (mainly luteinized stromal cells) in the majority of cases. Dimeric activin A was produced by all of the three OET cell lines with a 1.5-1.9-fold increment after FSH stimulation. However, activin A production was not augmented by LH treatment. No inhibin A was produced by the three OET cell lines with or without gonadotropin stimulation. CONCLUSIONS: The stroma of OET is the major source in the production of inhibin alpha (monomer). Dimeric inhibin A production may be the result of combined efforts of the tumor stroma (alpha subunit) and epithelium (betaA subunit). Cellular, compartmental expression of inhibin and activin subunits may play a role in the development of OET, although the mechanism remains undefined. The unopposed activin A production stimulated by FSH in OET cell lines suggests that activin production may represent one of the cellular mechanisms of growth promotion by FSH.

Activins↗
PubMed2000

An iterative approach to the beam hardening correction in cone beam CT.

In computed tomography (CT), the beam hardening effect has been known to be one of the major sources of deterministic error that leads to inaccuracy and artifact in the reconstructed images. Because of the polychromatic nature of the x-ray source used in CT and the energy-dependent attenuation of most materials, Beer's law no longer holds. As a result, errors are present in the acquired line integrals or measurements of the attenuation coefficients of the scanned object. In the past, many studies have been conducted to combat image artifacts induced by beam hardening. In this paper, we present an iterative beam hardening correction approach for cone beam CT. An algorithm that utilizes a tilted parallel beam geometry is developed and subsequently employed to estimate the projection error and obtain an error estimation image, which is then subtracted from the initial reconstruction. A theoretical analysis is performed to investigate the accuracy of our methods. Phantom and animal experiments are conducted to demonstrate the effectiveness of our approach.

Algorithms↗
PubMed2000

Recognition and silencing of repeated DNA.

Mechanisms for repetition of DNA pose both opportunities and challenges to a functional genome: opportunities for increasing gene expression by amplification of useful sequences, and challenges of controlling amplification by unwanted sequences such as transposons and viruses. Experiments in numerous organisms have suggested the likely existence of a general mechanism for recognition of repeated character in DNA. This review focuses (a) on the nature of these recognition mechanisms, and (b) on types of chromatin modification and gene silencing that are used to control repeated DNA.

Animals↗
PubMed1999

The RING finger/B-box factor TAM-1 and a retinoblastoma-like protein LIN-35 modulate context-dependent gene silencing in Caenorhabditis elegans.

Context-dependent gene silencing is used by many organisms to stably modulate gene activity for large chromosomal regions. We have used tandem array transgenes as a model substrate in a screen for Caenorhabditis elegans mutants that affect context-dependent gene silencing in somatic tissues. This screen yielded multiple alleles of a previously uncharacterized gene, designated tam-1 (for tandem-array-modifier). Loss-of-function mutations in tam-1 led to a dramatic reduction in the activity of numerous highly repeated transgenes. These effects were apparently context dependent, as nonrepetitive transgenes retained activity in a tam-1 mutant background. In addition to the dramatic alterations in transgene activity, tam-1 mutants showed modest alterations in expression of a subset of endogenous cellular genes. These effects include genetic interactions that place tam-1 into a group called the class B synMuv genes (for a Synthetic Multivulva phenotype); this family plays a negative role in the regulation of RAS pathway activity in C. elegans. Loss-of-function mutants in other members of the class-B synMuv family, including lin-35, which encodes a protein similar to the tumor suppressor Rb, exhibit a hypersilencing in somatic transgenes similar to that of tam-1 mutants. Molecular analysis reveals that tam-1 encodes a broadly expressed nuclear protein with RING finger and B-box motifs.

Amino Acid Sequence↗
PubMed1999

A two-site kinetic mechanism for ATP binding and hydrolysis by E. coli Rep helicase dimer bound to a single-stranded oligodeoxynucleotide.

Escherichia coli Rep helicase catalyzes the unwinding of duplex DNA in reactions that are coupled to ATP binding and hydrolysis. We have investigated the kinetic mechanism of ATP binding and hydrolysis by a proposed intermediate in Rep-catalyzed DNA unwinding, the Rep "P2S" dimer (formed with the single-stranded (ss) oligodeoxynucleotide, (dT)16), in which only one subunit of a Rep homo-dimer is bound to ssDNA. Pre-steady-state quenched-flow studies under both single turnover and multiple turnover conditions as well as fluorescence stopped-flow studies were used (4 degrees C, pH 7.5, 6 mM NaCl, 5 mM MgCl2, 10 % (v/v) glycerol). Although steady-state studies indicate that a single ATPase site dominates the kinetics (kcat=17(+/-2) s-1; KM=3 microM), pre-steady-state studies provide evidence for a two-ATP site mechanism in which both sites of the dimer are catalytically active and communicate allosterically. Single turnover ATPase studies indicate that ATP hydrolysis does not require the simultaneous binding of two ATP molecules, and under these conditions release of product (ADP-Pi) is preceded by a slow rate-limiting isomerization ( approximately 0.2 s-1). However, product (ADP or Pi) release is not rate-limiting under multiple turnover conditions, indicating the involvement of a second ATP site under conditions of excess ATP. Stopped-flow fluorescence studies monitoring ATP-induced changes in Rep's tryptophan fluorescence displayed biphasic time courses. The binding of the first ATP occurs by a two-step mechanism in which binding (k+1=1.5(+/-0.2)x10(7) M-1 s-1, k-1=29(+/-2) s-1) is followed by a protein conformational change (k+2=23(+/-3) s-1), monitored by an enhancement of Trp fluorescence. The second Trp fluorescence quenching phase is associated with binding of a second ATP. The first ATP appears to bind to the DNA-free subunit and hydrolysis induces a global conformational change to form a high energy intermediate state with tightly bound (ADP-Pi). Binding of the second ATP then leads to the steady-state ATP cycle. As proposed previously, the role of steady-state ATP hydrolysis by the DNA-bound Rep subunit may be to maintain the DNA-free subunit in an activated state in preparation for binding a second fragment of DNA as needed for translocation and/or DNA unwinding. We propose that the roles of the two ATP sites may alternate upon binding DNA to the second subunit of the Rep dimer during unwinding and translocation using a subunit switching mechanism.

Adenosine Diphosphate↗