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Biomedical subjects

H S Ko

Publications and source records attributed to H S Ko.

At least 19 recordsLinked to original sources

[Negative endoscopy and MSCT findings in patients with acute lower gastrointestinal hemorrhage. Value of (99m)Tc erythrocyte scintigraphy].

PURPOSE: To evaluate the clinical use of (99m)Tc red blood cell imaging (RBC imaging) in patients presenting with acute lower gastrointestinal (GI) bleeding and negative endoscopy and multislice computed tomography (MSCT) findings. PATIENTS AND METHODS: In 31 consecutive patients with acute lower GI bleeding in whom the endoscopy findings were negative or the procedure was not feasible, dual-phase MSCT of the abdomen was performed [collimation 4x1 mm (arterial phase), 4x2.5 mm (venous phase)]. MSCT was followed by a (99m)Tc red blood cell scan in patients in whom no active bleeding was visible by CT. Images were created within 24 h after administration of the tracer, depending on the clinical symptoms. The results of the imaging modalities were correlated with clinical course and surgical treatment. RESULTS: In 20 of 31 patients MSCT showed no active bleeding and a (99m)Tc red blood cell scan was performed. In 8 of 20 patients RBC imaging was also negative. Of these eight patients five were stable and did not require further diagnostic work-up; in the other three bleeding persisted and these patients required surgical treatment. In 12 of 20 patients active bleeding was demonstrated using a (99m)Tc red blood cell scan. Of 12 patients with positive RBC scintigraphy findings, 8 underwent surgery, where the site of bleeding was confirmed. CONCLUSION: In patients with acute lower GI bleeding with negative or nondiagnostic endoscopy or MSCT findings, (99m)Tc red blood cell imaging is a useful tool in an emergency algorithm, improving the overall bleeding detection rate.

Acute Disease↗

Mitochondrial DNA content is decreased in autosomal dominant optic atrophy.

BACKGROUND: Autosomal dominant optic atrophy (ADOA) is the commonest form of inherited optic neuropathy. Mutations in the OPA1 gene encoding a dynamin-related mitochondrial protein underlie ADOA and may perturb the biogenesis and maintenance of mitochondria. OBJECTIVE: To investigate the mutation spectrum of the OPA1 gene and assess alterations in mitochondrial content caused by OPA1 mutations. METHODS: Sixteen Korean patients with clinically suspected ADOA were studied. The mutation spectrum of the OPA1 gene was analyzed by PCR single-strand conformation polymorphism and sequencing, and mitochondrial DNA (mtDNA) content was quantified by real-time PCR. RESULTS: Eight different mutations were found, including five novel mutations. Quantitative real-time PCR analysis showed excellent linearity and precision for the determination of mtDNA copy numbers. The number of mtDNA copies per cell in patients with OPA1 gene mutations (ages 7 to 40) was significantly lower than those in all normal control subjects (p = 0.037), particularly lower than in normal control subjects ages 10 to 39 (p = 0.022). CONCLUSION: The mutation spectrum of the OPA1 gene disclosed marked genetic heterogeneity and the mitochondrial DNA content was found to be lower in autosomal dominant optic neuropathy, which provides direct evidence for a pathogenetic role of mutations of the OPA1 gene.

Adolescent↗

[MRI-morphology and staging of congenital mesoblastic nephroma: evaluation of a collection with 20 patients].

PURPOSE: To differentiate classic and cellular type of congenital mesoblastic nephroma (CMN) in MRI and to evaluate MRI for staging according to the Societe Internationale de Oncologie Pediatrique (SIOP). MATERIAL AND METHODS: MRI examinations of 20 children with CMN (age 1st to 16th months, classic type n = 11, cellular type n = 7, mixed type n = 2) were analyzed retrospectively. Cysts, necrosis, hemmorhage in the tumor, signal intensity, tumor structure, thrombosis and dilatation of renal vein, crossing of the body midline, peripheral contrast-enhancement, tumor volume and existence of a tumor pseudocapsule in contrast to the residual kidney were described. The radiologic stage was compared with the histopathologic stage (infiltration of perirenal fat and infiltration of the renal sinus). RESULTS: Tumors of the classic type (mean volume 67.9 ml) had necrosis in 1 case, crossed the midline in 1 case, had no cysts or bleeding, and had a peripheral contrast-enhancement in 1 case, and were heterogeneous in 9 cases. The cellular type (mean volume 302.8 ml) had tumor necrosis in 6 cases, bleeding in 3 cases, cysts in 3 cases, crossed the midline in 4 cases, and peripheral contrast enhancement in 2 cases, and was predominantly heterogeneous. Mixed tumor types (7 ml and 202 ml) had tumor necrosis in 1 case and crossed the midline in 1 case, a peripheral contrast enhancement in 2 cases and a homogenous structure in 1 case. The signal intensity in T1 w and T2 w images was not specific. The renal vein was inconspicuous in all children. The evaluation of the infiltration in perirenal fatty tissue was true positive in 1 case, true negative in 10 cases, false negative in 4 cases and false positive in 5 cases. The infiltration of the renal hilus was true positive in 10 children, false positive in 8 cases and true negative in 2 cases. CONCLUSION: A typical finding of CMN in MRI is a heterogeneous tumor without demarcation from the rest of the kidney parenchyma by a pseudocapsule. The cellular type of CMN tends to have a higher tumor volume and shows more necrosis, bleeding and cysts than the classic type in MRI. A peripheral contrast-enhancement in MRI is not characteristic for any type of CMN. Local tumor staging is not possible with MRI.

Diagnosis, Differential↗

[Localization of bleeding using 4-row detector-CT in patients with clinical signs of acute gastrointestinal hemorrhage].

PURPOSE: There is no gold-standard regarding the diagnostic work-up and therapy of an acute gastrointestinal (GI) hemorrhage. In most cases endoscopy provides the diagnosis but in a low percentage this modality is not feasible or negative. Purpose of this study was to evaluate the role of multi-phase Multi-Slice-Computertomography (MSCT) as a modality to diagnose and locate the site of acute GI hemorrhage in case of unfeasible or technically difficult endoscopy. MATERIALS AND METHODS: 58 patients, presenting with clinical signs of lower GI hemorrhage, were examined through a 24-month period. Preliminary endoscopy was either negative or unfeasible. Images were obtained with a four-detector row CT with an arterial (4 x 1 mm collimation, 0.8 mm increment, 1.25 mm slice width, 120 kV, 165 mAs) and portal venous series (4 x 2,5 mm collimation, 2 mm increment, 3 mm slice width, 120 kV, 165 mAs). Time interval between endoscopy and CT varied between 30 minutes and 3 hours. The results of the MSCT were correlated with clinical course and surgical or endoscopical treatment. RESULTS: 20 of the 58 patients (34 %) undergoing MSCT had a bleeding site identified, thus providing decisive information for the following intervention. In case of a following therapeutic intervention there was 100 % correlation regarding the bleeding site. In 38 of the 58 patients (66 %), a bleeding site was not identified by MSCT. Twenty of these 38 patients (53 %) were stable and required no further treatment. In 18 of these 38 patients further interventional therapy was required due to continuing hemorrhage and in all of those patients the bleeding site was detected by intervention. CONCLUSION: Compared to other diagnostic methods MSCT is a fast, widely-available and low-risk technique for the localization of active GI hemorrhage. The clinical use seems to be justified since in more than one third of the patients, MSCT demonstrates the site of bleeding and provides decisive information for further interventional therapy. Concerning those patients, in whom MSCT is negative (38 out of 58 patients), only every second patient requires any additional diagnostic work-up.

Acute Disease↗

Head-column field-amplified sample stacking in capillary electrophoresis for the determination of cimetidine, famotidine, nizatidine, and ranitidine-HCl in plasma.

In this study, low concentrations of histamine2-receptor (H2-)antagonists were effected across a water plug, with separation taking place in a binary buffer comprising ethylene glycol and NaH2PO4 (pH 5.0), and detection at 214 nm. Liquid-liquid extraction with ethyl acetate- isopropanol is shown to provide extracts that are sufficiently clean. The calibration curves were linear over a concentration range of 0.1-2.00 microg/mL cimetidine, 0.2-5.0 microg/mL ranitidine-HCl, 0.3-5.0 microg/mL nizatidine, and 0.1-3.0 microg/mL famotidine. Mean recoveries were > 82%, while the intra- and interday relative standard deviations (RSDs) and relative errors (REs) were all < 13%. The method is sensitive with a detection limit of 3 ng/mL cimetidine, 30 ng/mL ranitidine HCl, 50 ng/mL nizatidine and 10 ng/mL famotidine (S/N = 3, electric-driven injection 90 s). This newly developed capillary electrophoresis (CE) method was applied for the determination of analytes extracted from plasma taken from a volunteer dosing a cimetidine, ranitidine, and nizatidine tablet simultaneously. These three H2-antagonists can be detected in real samples by this method, excluding the low dosing of famotidine tablet.

Buffers↗

Simultaneous determination of cimetidine, famotidine, nizatidine, and ranitidine in tablets by capillary zone electrophoresis.

A simple capillary zone electrophoresis (CZE) method is described for the simultaneous determination of cimetidine (CIM), famotidine (FAM), nizatidine (NIZ), and ranitidine (RAN). The analysis of these drugs was performed in a 100 mM phosphate buffer, pH 3.5. Several parameters were studied, including wavelength for detection, concentration and pH of phosphate buffer, and separation voltage. The quantitative ranges were 100-1,000 microM for each analyte. The intra- and interday relative standard deviations (n = 5) were all less than 4%. The detection limits were found to be about 10 microM for CIM, 20 microM for RAN, 20 microM for NIZ, and 10 microM for FAM (S/N = 3, injection 1 s) at 214 nm. All recoveries were greater than 92%. Applications of the method to the assay of these drugs in tablets proved to be feasible.

Anti-Ulcer Agents↗

Hypoxia decreases proteins involved in epithelial electrolyte transport in A549 cells and rat lung.

Fluid reabsorption from alveolar space is driven by active Na reabsorption via epithelial Na channels (ENaCs) and Na-K-ATPase. Both are inhibited by hypoxia. Here we tested whether hypoxia decreases Na transport by decreasing the number of copies of transporters in alveolar epithelial cells and in lungs of hypoxic rats. Membrane fractions were prepared from A549 cells exposed to hypoxia (3% O(2)) as well as from whole lung tissue and alveolar type II cells from rats exposed to hypoxia. Transport proteins were measured by Western blot analysis. In A549 cells, alpha(1)- and beta(1)-Na-K-ATPase, Na/K/2Cl cotransport, and ENaC proteins decreased during hypoxia. In whole lung tissue, alpha(1)-Na-K-ATPase and Na/K/2Cl cotransport decreased. alpha- and beta-ENaC mRNAs also decreased in hypoxic lungs. Similar results were seen in alveolar type II cells from hypoxic rats. These results indicate a slow decrease in the amount of Na-transporting proteins in alveolar epithelial cells during exposure to hypoxia that also occurs in vivo in lungs from hypoxic animals. The reduced number of transporters might account for the decreased transport activity and impaired edema clearance in hypoxic lungs.

Adenocarcinoma, Bronchiolo-Alveolar↗

Total hip reconstruction in the anatomically distorted hip. Cemented versus hybrid total hip arthroplasty.

Recent reports in the literature strongly support the idea that cement is the optimum form of fixation of the femoral component in total hip replacement. For hybrid total hip arthroplasty, we used a cemented cup instead of an uncemented cup since this was inevitable in cases of poorly developed acetabulum. The uncemented cone femoral component is also beneficial in cases of extremely narrow and cylindrical configuration of the medullary cavity of untreated congenital dislocation of the hips (CDH) and tuberculosis or septic arthritis in childhood. We reviewed the clinical result of a consecutive series of patients with cemented total hip arthroplasty (THA) compared with recent studies on hybrid reconstruction using survivorship analysis. This subsequent study involved a hybrid uncemented Wagner cone femoral component and a cemented acetabular component with roof reinforcement by additional impacted cancellous allograft with hydroxyapatite (HA). We believe that early failure of the cemented components was due to an adverse effect of thin cement mantles around cemented femoral stems as well as the cemented cup in THA. In addition, 8 patients who received our modification of the Charnley CDH component had poor results even though we reduced the stem geometry and thickness. Furthermore, intraoperative fracture and splitting of the proximal femur was a major complication during implantation. Contrary to expectations, the results of these hybrid reconstructions were extremely encouraging (average follow-up period of 2-4.5 years). In addition to our experience of the cemented versus hybrid THA involving the uncemented Wagner femoral component, we have determined positive indications for the untreated and severely distorted anatomy of bilateral CDH for which surgical interventions for reconstruction were not recommended before.

Adult↗

A rapid and sensitive cellular enzyme-linked immunoabsorbent assay (CELISA) for the detection and quantitation of antibodies against cell surface determinants. I. A comparison of cell fixation and storage techniques.

A solid phase cellular ELISA was designed and evaluated for the detection of antibodies specific for cell surface determinants. It was hypothesized that certain fixation and freezing procedures would result in stabilization of cell structures for prevention of antigen diffusion and extraction during washing procedures. This would assure assay accuracy and convenient sample management. It was hypothesized that fixation with certain reagents prior to analysis would not alter antigenicity of antibody targeted epitopes. In order to improve the preservation of the cells following cell binding to the solid phase matrix while still retaining antigenicity and morphology, a series of fixatives and storage procedures were screened to determine which were best suited for CELISA. Methanol, washing buffer (WB), Hanks' balanced salt solution (HBSS), and 0.5% formalin in HBSS were examined by comparing their relative cell binding capacity and the subsequent cell morphology. In consideration of all variables, fixation in 0.5% formalin provided the best maintenance of cell antigenicity, morphology, binding, and was associated with consistent results. Cells used immediately after fixation and fixed cells used after storage at -80 degrees C for up to 12 months were compared to determine if long term storage affected antigenicity. Since frozen cells and fresh cells demonstrated statistically identical positive to negative ratios and consistency of antibody binding, it was determined that long term frozen storage of formalin-fixed cells did not adversely affect antibody binding capacity to cell surface determinants.

Animals↗

Induction of the POU domain transcription factor Oct-2 during T-cell activation by cognate antigen.

Oct-2 is a transcription factor that binds specifically to octamer DNA motifs in the promoters of immunoglobulin and interleukin-2 genes. All tumor cell lines from the B-cell lineage and a few from the T-cell lineage express Oct-2. To address the role of Oct-2 in the T-cell lineage, we studied the expression of Oct-2 mRNA and protein in nontransformed human and mouse T cells. Oct-2 was found in CD4+ and CD8+ T cells prepared from human peripheral blood and in mouse lymph node T cells. In a T-cell clone specific for pigeon cytochrome c in the context of I-Ek, Oct-2 was induced by antigen stimulation, with the increase in Oct-2 protein seen first at 3 h after activation and continuing for at least 24 h. Oct-2 mRNA induction during antigen-driven T-cell activation was blocked by cyclosporin A, as well as by protein synthesis inhibitors. These results suggest that Oct-2 participates in transcriptional regulation during T-cell activation. The relatively delayed kinetics of Oct-2 induction suggests that Oct-2 mediates the changes in gene expression which occur many hours or days following antigen stimulation of T lymphocytes.

Animals↗

Mechanism of bcl-2 activation in human follicular lymphoma.

The t(14; 18) chromosomal translocation of human follicular lymphoma recombines the bcl-2 gene from chromosome 18 with the immunoglobulin heavy chain joining region. In the t(14; 18) translocation bearing cell line SU-DHL-6, this results not only in an inappropriately high rate of bcl-2 transcription for a mature B cell, but also in two potentially critical point mutations. To determine the relative importance of these mutations, we searched for their presence in DNA from the involved lymph nodes of 12 patients with t(14; 18) follicular lymphoma. bcl-2 genomic sequences were specifically amplified by the polymerase chain reaction technique and then directly sequenced. None of the 12 samples analysed revealed the codon 7 or codon 129 mutation detected in SU-DHL-6. We conclude that abnormal expression of bcl-2 rather than structural alterations at codon 7 or 129 play an important role in the disordered growth and differentiation of follicular B-cell lymphoma.

B-Lymphocytes↗

A human protein specific for the immunoglobulin octamer DNA motif contains a functional homeobox domain.

The homeobox domain is shared by Drosophila homeotic proteins, yeast mating type proteins, and some functionally uncharacterized mammalian proteins. A lymphoid-restricted human protein that binds to the immunoglobulin octamer regulatory motif was shown to contain an amino acid sequence that has 33% amino acid identity with the consensus sequence of the previously cloned homebox domains. This homeobox gene was localized to chromosome 19, thus mapping separately from other human homebox genes. A mutant protein containing amino acid substitutions within a putative helix-turn-helix motif in the homeobox domain did not bind DNA detectably. This human homeobox protein was shown to bind the same DNA sequence as the homeobox domains of the yeast mating type proteins and Drosophila homeotic protein, suggesting that homeobox proteins may have closely related DNA binding characteristics.

Amino Acid Sequence↗

Consequences of the t(14;18) chromosomal translocation in follicular lymphoma: deregulated expression of a chimeric and mutated BCL-2 gene.

The t(14;18) chromosomal translocation of human follicular lymphoma recombines the candidate transforming gene bcl-2, located at 18q21, with the immunoglobulin (Ig) H-chain joining region (JH) at 14q32. To elucidate the consequences of this translocation, we cloned bcl-2 cDNAs from a pre-B cell line (Nall-1) and a t(14;18) lymphoma cell line (SU-DHL-6) and compared these sequences with their genomic counterparts. These studies revealed the complexity of bcl-2 gene expression in which six potential polyadenylation signals in exon 3 and two different 5' exons (exons 1 and 2) and promoters are alternatively used to generate different sized bcl-2 mRNAs. A single open reading frame (ORF), at the junction of exons 2 and 3, predicts a 239 amino acid, 26 kD protein. Most chromosome 18 breakpoints cluster within a 150 bp region of exon 3. In SU-DHL-6 the t(14;18) translocation juxtaposes a truncated bcl-2 gene with J6 in a tail-to-head configuration, resulting in the deregulated expression of chimeric bcl-2/Ig transcripts. Importantly, the SU-DHL-6 bcl-2 cDNA also contained several point mutations in the ORF, two of which altered the primary amino acid sequence. The deregulated expression of an altered bcl-2 gene may play a critical role in the disordered growth and differentiation of follicular B cell lymphoma.

Amino Acid Sequence↗

IgG and IgE antibody response following Pollinex-R immunotherapy.

Specific antibodies to ragweed pollen antigens were studied in 22 patients given preseason Pollinex-R (glutaraldehyde-modified ragweed tyrosine-adsorbate) (PR), 17 without immunotherapy, and 8 on maintenance doses of aqueous ragweed extracts. The PR-group showed about fourfold increases in IgG antibodies in season when compared with pretreatment levels (P less than .001). IgG antibody changes from before season to season in other groups and IgE antibody changes in all three groups were not significant. Despite this, IgG antibodies in season had fallen from their peak achieved with PR. About one-third of PR-treated patients still manifested significant symptomatology irrespective of changes in IgG antibodies. We thus conclude that although changes in specific IgG may be an important correlate in PR-immunotherapy, additional mechanisms for clinical responsiveness remain to be clarified.

Adolescent↗

Serum thymosin-alpha 1: lack of association between elevated levels and HIV infection.

Serum thymosin-alpha 1 levels as detected by radioimmunoassay (RIA) have been reported to be elevated in AIDS. We studied 143 individuals in two risk groups for AIDS (male homosexuals and haemophiliacs) for serum thymosin-alpha 1 and antibodies to HIV. RIA for thymosin-alpha 1 was performed in Dr A. Goldstein's laboratory at George Washington University, Washington DC. We found that similar proportions of seropositive and seronegative subjects within each risk group showed elevated thymosin-alpha 1 levels. Retesting after 6 months did not reveal significant increments over previous levels in seropositive subjects or in those developing HIV antibodies in the repeat samples. Thus, although elevated thymosin-alpha 1 levels might be a possible serum marker for AIDS, their association with HIV infection was not demonstrated. In addition, the male homosexuals we studied showed significantly lower thymosin-alpha 1 levels than haemophiliacs but this difference remains to be explained.

Acquired Immunodeficiency Syndrome↗

Evidence for a defect in "switch" T cells in patients with immunodeficiency and hyperimmunoglobulinemia M.

Immunodeficiency with hyperimmunoglobulinemia M is a syndrome characterized by normal to elevated serum levels of IgM and low levels or absence of IgG and IgA. The defect in this syndrome is thought to reside within the B lymphocyte, which may be unable to undergo a "switch" in immunoglobulin class from IgM to IgG or IgA. To address this question more directly, we cultured B cells from nine patients with this syndrome with pokeweed mitogen and either "switch" T cells or normal control T cells. In cultures with normal T cells, only IgM was secreted, whereas in cultures with switch T cells, IgG as well as IgM, or IgM, IgG, and IgA were secreted. Furthermore, analysis of the immunoglobulin heavy-chain genes in these B cells by means of genetic probes of constant and switch regions revealed normal gene patterns. These data suggest that B cells from patients with hyperimmunoglobulinemia M may not be abnormal, as previously proposed, and that, at least in some patients with this syndrome, a defect in switch T cells may be pathogenic.

Adolescent↗

Responsiveness of human T-lymphocyte subpopulations in autologous mixed-lymphocyte reaction using xenoprotein-free separated cells: autologous reactivity lies chiefly in a low density T-cell fraction.

The responsiveness in the autologous mixed-lymphocyte reaction (AMLR) by human T cells separated using two different methods not involving xenoantigen contract was examined. Although T cells from nylon-wool columns were active in AMLR, T cells separated by a Percoll gradient method responded poorly. Further separation of T cells from nylon-wool columns into low density (TL) and high density (TH) fractions by Percoll revealed that TL was enriched, while TH was depleted, in AMLR responsiveness. This difference could not be accounted for by differences in the helper or suppressor cells in the fractions. Moreover, TH responded well in secondary AMLR. Therefore the T cells reactive in AMLR reside chiefly, although not exclusively, in the low density fraction.

Cell Separation↗