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Biomedical subjects

J S Ryu

Publications and source records attributed to J S Ryu.

At least 55 records · Page 3Linked to original sources

Biological and biochemical modulation of Trichomonas vaginalis KT9 isolate after shifting of culture medium from TPS-1 into TYM.

To evaluate the biological and biochemical characteristics of Trichomonas vaginalis KT9 isolate, the growth and size of trichomonads, pathogenicity in mouse, protein profiles and proteinase activity were examined after shifting the medium from TPS-1 into TYM. Generation time of trichomonads in TYM medium was 4.5 hr in comparison to TPS-1 with 7.1 hr. Size of trichomonads cultured in TPS-1 medium (8.5 +/- 0.9 x 6.0 +/- 0.9 microns) was significantly smaller than those in TYM medium (10.9 +/- 1.4 x 8.2 +/- 0.9 microns). Trichomonads cultured in TYM medium produced subcutaneous abscess in 9 out of 10 mice, whereas those in TPS-1 medium produced abscesses in 2 out of 10 mice. In SDS-PAGE, trichomonad lysates from both media showed ten common bands. However, trichomonads in TYM medium showed additional bands of 136 kDa, 116 kDa and 40 kDa in comparison to those in TPS-1 with 100 kDa. By immunoblot with T. vaginalis-immunized rabbit sera, T. vaginalis cultivated in both TYM and TPS-1 media showed 5 common bands, and unique bands of 116 kDa, 105 kDa, and 86 kDa were observed in trichomonads in TYM while a 140 kDa band in those in TPS-1. In gelatin SDS-PAGE, trichomonads in TYM degraded gelatin stronger than those in TPS-1. Also protease activity of trichomonads in TYM was significantly higher than that of trichomonads in TPS-1 using Bz-Pro-Phe-Arg-Nan as a substrate. According to the results, it is assumed that the shift from TPS-1 into TYM medium for cultivation of T. vaginalis might modulate the biological and biochemical properties of T. vaginalis in vitro.

Animals↗

Degradations of human immunoglobulins and hemoglobin by a 60 kDa cysteine proteinase of Trichomonas vaginalis.

The present study was undertaken to investigate the role of cysteine proteinase of Trichomonas vaginalis in escaping from host defense mechanism. A cysteine proteinase of T. vaginalis was purified by affinity chromatography and gel filtration. Optimum pH for the purified proteinase activity was 6.0. The proteinase was inhibited by cysteine and serine proteinase inhibitors such as E-64, NEM, IAA, leupeptin, TPCK and TLCK, and also by Hg2+, but not affected by serine-, metallo-, and aspartic proteinase inhibitors such as PMSF, EDTA and pepstatin A. However, it was activated by the cysteine proteinase activator, DTT. The molecular weight of a purified proteinase was 62 kDa on gel filtration and 60 kDa on SDS-PAGE. Interestingly, the purified proteinase was able to degrade serum IgA, secretory IgA, and serum IgG in time- and dose-dependent manners. In addition, the enzyme also degraded hemoglobin in a dose-dependent manner. These results suggest that the acidic cysteine proteinase of T. vaginalis may play a dual role for parasite survival in conferring escape from host humoral defense by degradation of immunoglobulins, and in supplying nutrients to parasites by degradation of hemoglobin.

Animals↗

Aberrant cervical thymus: a case report and review of literature.

This article illustrates a case of an aberrant cervical thymus presented as a neck mass. This is a case of a 4-month-old boy presenting with a right submandibular mass whose preoperative diagnosis was lymphangioma or neoplastic lesion. The mass was successfully removed and the histopathological examination showed normal thymic tissue with no diagnostic abnormality. This paper reviews the embryological background of aberrant cervical thymus, the varying clinical presentations with an emphasis on differential diagnosis, clinical work-up, and surgical treatment.

Choristoma↗

Resistance to tobamoviruses in transgenic tobacco plants expressing the coat protein gene of pepper mild mottle virus (Korean isolate).

Red pepper, one of the most important vegetable crops in Korea, is severely affected by viral diseases causing 20-50% reduction in product yield. A pepper strain of tobacco mosaic virus (TMV-p) is the most common virus in red pepper. To study the molecular structure of the TMV-p virus, we generated cDNA clones of the viral genome. Partial sequencing of a few cDNA clones revealed that TMV-p shares a 98% identity at the nucleotide level with the Spanish isolate of pepper mild mottle virus (PMMV-s). This suggests that TMV-p should be reclassified as the Korean isolate of PMMV (PMMV-k). The coat protein (CP) gene together with the 3' untranslated region of the PMMV-k virus was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) using oligomers deduced from the sequence of PMMV-s. The sequence analysis of the CP gene and the 3' untranslated region further confirmed that PMMV-k is highly related to PMMV-s. The CP gene and the 3' untranslated region of PMMV-k were cloned into a plant expression vector and the construct was introduced into tobacco plants. The transgenic plants expressing the PMMV-k CP gene were delayed in developing systemic disease or failed to develop symptoms at all after inoculation with PMMV-k. Delay of symptoms was also observed when the plants were inoculated with TMV-OM which shares a 74% homology with PMMV-k in the amino acid sequence of the CP region. In a local lesion host, the CP expressing plants exhibited a greatly reduced number of necrotic lesions as compared to control plants after inoculation with TMV-OM. Our results show that CP-mediated viral resistance is readily applicable in the case of PMMV-k and can provide resistance to other viruses in the tobamovirus group.

Amino Acid Sequence↗

Cloning of a cysteine proteinase cDNA of adult Paragonimus westermani by polymerase chain reaction.

Cysteine proteinase cDNA fragment from adult mammalian trematode, Paragonimus westermani was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved cysteine proteinase sequences. The 5' and the 3' regions of the cysteine proteinase gene were amplified using the PCR protocol for the rapid amplification of cDNA ends (RACE). It has an open reading frame of 804 bp. The deduced amino acid sequence consists of 268 amino acids. Sequence analysis and alignment showed significant sequence similarity to other eukaryotic cysteine proteinases and conservation of the cysteine, histidine, and asparagine residues that form the catalytic triad. The cysteine proteinase cDNA fragment was also subcloned in the expression vector pET and expressed as a C-terminal His-tag fusion protein in Escherichia coli.

Amino Acid Sequence↗

Diagnosis of orbital cavernous hemangioma with Tc-99m RBC SPECT.

The authors report two cases of orbital cavernous hemangioma diagnosed by Tc-99m RBC SPECT. Tc-99m RBC SPECT showed a typical scintigraphic pattern commonly seen in hepatic hemangioma in which there is intense focally increased uptake on delayed SPECT images. Tc-99m RBC SPECT in orbital cavernous hemangioma may be as useful a diagnostic modality as in hepatic hemangioma.

Adult↗

Degradation of human immunoglobulins and cytotoxicity on HeLa cells by live Trichomonas vaginalis.

The present study was undertaken to determine whether live T. vaginalis degrades human secretory IgA, serum IgA and IgG molecules. Human immunoglobulins were exposed to live trophozoites, parasite lysate, and excretory-secretory product (ESP) of T. vaginalis. To determine the fragmentation of immunoglobulins, the reaction sample was subjected to SDS-PAGE and EITB, and peroxidase conjugated antihuman IgA and IgG were used as probes. Live trophozoites degraded secretory IgA. Serum IgA and IgG, and degradation were pressed forward by the prolongation of the incubation time and by increasing the number of trichomonads respectively. Also the lysates and ESP of trichomonads degraded IgA and IgG. The cysteine and serine proteinase inhibitors such as E-64, antipain, iodoacetic acid, iodoacetamide, TLCK reduced the ability of cleaving immunoglobulins. The proteinase activity and cytotoxicity of T. vaginalis to HeLa cells were decreased when live T. vaginalis was treated with metallo-proteinase inhibitor as well as cysteine and serine proteinase inhibitors. These results suggest that proteinase secreted from live T. vaginalis may play a part role in host pathogenesis by T. vaginalis, and the cleaving ability of host immunoglobulins by the proteinase may contribute as a one of immune evasion mechanism for parasite survival in the host.

Animals↗

The role of nitric oxide as an effector of macrophage-mediated cytotoxicity against Trichomonas vaginalis.

The purpose of this study is to determine whether nitric oxide is involved in the extracellular killing of Trichomonas vaginalis by mouse (BALB/c) peritoneal macrophages and RAW264.7 cells activated with LPS or rIFN-gamma and also to observe the effects of various chemicals which affect the production of reactive nitrogen intermediates (RNI) in the cytotoxicity against T. vaginalis. The cytotoxicity was measured by counting the release of [3H]-thymidine from labelled protozoa and NO2- was assayed by Griess reaction. NG-monomethyl-L-arginine (L-NMMA), NG-nitro-L-arginine methyl ester (NAME) and arginase inhibited cytotoxicity to T. vaginalis and nitrite production by activated mouse perioneal macrophages and RAW 264.7 cells. The addition of excess L-arginine competitively restored trichomonacidal activity of macrophages. Exogenous addition of FeSO4 inhibited cytotoxicity to T. vaginalis and nitric products of macrophages. From above results, it is assumed that nitric oxide plays an important role in the host defense mechanism of macrophages against T. vaginalis.

Animals↗

Imaging and quantitation of dopamine transporters with iodine-123-IPT in normal and Parkinson's disease subjects.

UNLABELLED: Iodine-123-N-(3-iodopropene-2-yl)-2beta-carbomethoxy-3beta-( 4-chlorophenyl) tropane (123I-IPT) is a new dopamine transporter ligand that selectively binds the dopamine reuptake sites. Transporter concentrations have been known to decrease in Parkinson's disease patients. The purpose of this study was to evaluate the usefulness of IPT as an imaging agent for measuring changes in transporter concentrations in Parkinson's disease. METHODS: IPT labeled with 6.78 +/- 0.67 mCi 123I was injected intravenously as a bolus into eight normal controls (mean age 41 +/- 12 yr) and 17 Parkinson's disease patients (mean age 55 +/- 9 yr). Dynamic SPECT scans of the brain were then performed for 5 min each over 120 min on a triple-headed gamma camera equipped with medium-energy collimators. Regions of interest were drawn on the middle set of the image at the level of the basal ganglia (BG) for each subject. Time-activity curves were generated for the left BG, right BG and occipital cortex (OCC). The empirical ratios between BG-OCC and OCC, which represent specific-to-nonspecific binding ratios, were computed at various time points. The statistical parameter k3/k4 was estimated by two methods: a variation of the graphic method that derives the ratio of ligand distribution volumes (R[V]) and the area ratio method (R[A]), in which the ratio is calculated from the areas under the specific and nonspecific binding activity curves. RESULTS: The mean (BG-OCC)/OCC ratio for normal controls (3.07 +/- 0.73) was significantly higher than that for Parkinson's disease patients at 115 min (1.10 +/- 0.56) (p = 2.76 x 10[-5]). The mean R(V) and R(A) for normal controls were 2.06 +/- 0.27 and 1.50 +/- 0.15, respectively. The mean R(V) and R(A) for Parkinson's disease patients were 0.78 +/- 0.31 and 0.65 +/- 0.24, respectively. Both R(V) and R(A) for normal controls were significantly higher than those for Parkinson's disease patients (p values for R(V) and R(A) were 1.91 x 10(-8) and 3.46 x 10(-10), respectively). The R(V) has linear relationships with both R(A) and (BG-OCC)/OCC ratio at 115 min. The R(V) has a higher correlation (r = 0.99) with R(A) than it does with (BG-OCC)/OCC (r = 0.93). CONCLUSION: The R(V), R(A) and (BG-OCC)/OCC for Parkinson's disease patients were clearly separated from those of normal controls, and they may be useful outcome measures for clinical diagnosis. The simplest (BG-OCC)/OCC ratio, requiring a single late time point, could be useful in clinical situations, whereas R(V) or R(A) is preferred when the dynamic data are available. The findings suggest that 123I-IPT is a useful tracer for diagnosing Parkinson's disease and studying dopamine reuptake sites.

Adult↗

Serial measurement of interleukin-6, transforming growth factor-beta, and S-100 protein in patients with acute stroke.

BACKGROUND AND PURPOSE: Cytokine changes in patients with acute stroke have been insufficiently studied. The purpose of this study was to delineate the characteristics of serial changes of serum interleukin-6 (IL-6) and transforming growth factor-beta (TGF-beta) in patients with cerebral infarction and intracerebral hemorrhage. METHODS: We serially (within 24 hours, at day 3, and at day 7) measured the serum levels of IL-6 and TGF-beta in 29 patients with acute stroke (10 with large cortical cerebral infarction, 9 with subcortical small infarction, and 10 with intracerebral hemorrhage). As an index of brain damage. S-100 protein was also measured. Twelve age-matched healthy subjects were tested as a control. RESULTS: S-100 protein was detected in only 11 patients with large infarction or intracerebral hemorrhage. Its level peaked at day 3 in patients with infarction, whereas it peaked within 24 hours in those with intracerebral hemorrhage. The level of IL-6 was most markedly elevated at day 1, which tended to decrease thereafter. However, its level remained significantly elevated compared with that of the control group even at day 7. The level of TGF-beta was significantly decreased at day 1 and day 3 and tended to return toward the control value thereafter. The levels of both cytokines were not significantly different among the three different stroke subtypes and were not correlated with the number of blood leukocytes and platelets. CONCLUSIONS: The alteration of IL-6 and TGF-beta levels, which occurs rapidly after acute stroke regardless of the subtype, may reflect the changing immunological-inflammatory status of these patients and does not appear to reflect merely the consequence of the brain damage.

Acute Disease↗

[Characterization of the partially purified proteinase from Trichomonasvaginalis].

Characterization of a purified proteinase from Trichomonas vaginalis was carried out using bacitracin-sepharose affinity chromatography. Trichomonas vaginalis KT-9 isolate was used as a source of enzyme study. Proteinase activity was determined using Bz-Pro-Phe-Arg-Nan as the substrate. Optimum pH for the purified proteinase activity was 7.0 and 6.0, 9.0 with DTT. Optimum temperature was 37 degrees C and isoelectric point was 7.2. Activity of this proteinase was inhibited by E-64, antipain, leupeptin, Hg2+ and Zn2+ and activated by DTT and cysteine. Activity of the purified proteinase was visualized by gelatin SDS-PAGE. The gelatinolytic activity of the purified proteinase was inhibited by E-64, antipain, leupeptin, and IAA, but not by PMSF and EDTA. On SDS-PAGE, the molecular weight of the purified proteinase was 60,000 daltons. Sera of rabbits infected with T. vaginalis reacted specifically in immunoblots with this proteinase. These results indicate that 60 kDa of purified proteinase was cysteine proteinase with antigenicity.

Animals↗

Toxoplasma antibody titers by ELISA and indirect latex agglutination test in pregnant women.

The seroepidemiologic studies on anti-Toxoplasma antibody titers were carried out using ELISA and indirect latex agglutination test. Among 899 sera prepared from pregnant women, 39 cases (4.3%) revealed positive reaction and 218 sera from middle school students showed 4 positive reaction (1.8%) by ELISA. By LAT (newly established by National Veterinary Research Institute. Korea), the sera of 7 pregnant women (0.8%) showed positive reaction. When 80 sera showing > or = 1:8 by LAT were used for comparing the results obtained from LAT and Toxotest-MT (Eiken Chemical Co., Japan), 7 cases and 8 sera were positive, respectively. All of 11 sera of proven toxoplasmosis patients showed positive reaction in both tests. Overall proportion of agreement between LAT kit and Toxotest-MT was 0.94 (kappa-index = 0.632, p < 0.011, and LAT was considered to be useful for the screening of toxoplasmosis.

Adolescent↗

Cell cytotoxicity of sodium nitrite, sodium nitroprusside and Roussin's black salt against Trichomonas vaginalis.

We have investigated the action of sodium nitrite and other nitrosyl complexes, such as sodium nitroprusside and Roussin's black salt, on the growth of metronidazole-sensitive and resistant strains of Trichomonas vaginalis and their hydrogenosomal enzymes. All three chemicals inhibited the growth of T. vaginalis: sodium nitrite at 8 mM, sodium nitroprusside at 1.2 mM and Roussin's black salt at 0.2 mM. Metronidazole-sensitive (KT9) and resistant (CDC85) isolates showed similar cytotoxicity against these molecules. Specific activities of pyruvate:ferredoxin oxidoreductase and hydrogenase and oxygen uptake rates were decreased in the T. vaginalis isolate treated with sodium nitrite and sodium nitroprusside. However, Roussin's black salt increased the specific activity of pyruvate:ferredoxin oxidoreductase or hydrogenase in CDC85 or KT9 cells and increased the oxygen uptake rate in the KT9 isolate.

Animals↗

Bone SPECT imaging of vertebral hemangioma correlation with MR imaging and symptoms.

To evaluate the bone SPECT findings of vertebral hemangioma and their relationship to symptoms and MRI findings, the authors reviewed planar and SPECT bone scans and MR images of 15 vertebral hemangioma in 10 patients. All planar bone scan images demonstrated normal findings, except one, SPECT images demonstrated normal findings in 11 vertebral hemangioma less than 3 cm in diameter. Three of four vertebral hemangioma that were 3 cm or larger showed abnormally increased or decreased uptake on SPECT images. There was a disparity between SPECT imaging and MRI.

Adult↗

[Effect of sodium nitrite on Trichomonas vaginalis].

We have investigated the action of sodium nitrite on the growth and morphologic changes of T. vaginalis and on the treatment of subcutaneous abscess by trichomonad in mice. Sodium nitrite inhibited the growth of metronidazole-sensitive KT9 isolate and metronidazole-resistant CDC85 strain of T. vaginalis as concentration of 6 mM and 10 mM, respectively. Intraperitoneal injection of sodium nitrite (70 micrograms, 100 micrograms, 130 micrograms/g body weight) did not reduce the size of abscess produced by subcutaneous inoculation of T. vaginalis in mice. T. vaginalis, treated with sodium nitrite at concentration giving about 50% inhibition of growth, showed fissures, many vacuoles and electron-translucent zone in the cytoplasm by transmission electron microscopy. In the case of CDC85 treated with 9 mM sodium nitrite, hydrogenosomal matrical change, destruction of hydrogenosomal membrane, autophagic vacuoles, disappearance of Golgi complex and polysome were notably observed. With above results, it is assumed that sodium nitrite inhibits the growth of metronidazole-sensitive and--resistant strains of T. vaginalis and induces the morphological changes of T. vaginalis although it does not affect in reducing of abscess size by T. vaginalis in mice.

Abscess↗

Clinical value of DMSA planar and single photon emission computed tomography as an initial diagnostic tool in adult women with recurrent acute pyelonephritis.

Routine DMSA scintigraphy, ultrasound (US) of the kidney, intravenous pyelography (IVP) and voiding cystoureterography (VCU) were performed in 27 consecutive adult women with recurrent acute pyelonephritis (APN) during a 12-month follow-up. Both planar and single photon emission computed tomography (SPECT) images were obtained for DMSA scan. DMSA scans were repeated in those patients with abnormal initial scan. DMSA-SPECT showed normal findings in 2, single renal cortical detect (RCD) in 9 and multiple RCD in 16 (including nonvisualization in 2). Of the 11 kidneys with normal findings or single RCD on DMSA-SPECT, only 1 (9%) showed vesicoureteral reflux (VUR) on VCU (grade I). A large proportion of those with multiple RCDs showed abnormal findings on IVP (44%, 7/16), US (38%, 6/16) or VCU (31%, 5/16); 63% in any of these three studies. 5 of 6 patients with VUR had multiple RCDs on DMSA-SPECT, and 3 of these 5 showed no abnormality on IVP or US. 7 patients who needed other managements besides initial standard antibiotic treatment had multiple RCDs on DMSA-SPECT. 15 normal women were also studied and showed normal DMSA-SPECT, US and IVP, in all cases. Follow-up DMSA-SPECT was done in 16 patients (7 with single RCD, 9 with multiple RCD). All 7 patients with single RCD showed improvement, in those with multiple RCDs improvement was observed in 2, no change in 7 on follow-up studies. We conclude: (1) DMSA-SPECT is a useful initial diagnostic tool in adult women with recurrent APN to identify patients who need more extensive radiological studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Identification of surface antigen of Trichomonas vaginalis.

Plasma membrane proteins of a Korean isolate of Trichomonas vaginalis HY-1 were fractionated for antigen analysis. Homogenates of T. vaginalis were fractionated by the differential centrifugation using sucrose step-gradient method. The interface layer from the 25%/45% sucrose was collected as a plasma membrane fraction and its purity was examined by transmission electron microscopy. The antigenicity of plasma membrane fraction was analysed by enzyme-linked immunoelectrotransfer blot technique with immune rabbit serum and compared with surface antigen labelled with N-hydroxysuccinimide-biotin. The fluffy fraction of 25%/45% sucrose interface was homogeneous and membrane particles were present as extended sheet and concentric vesicles showing typical trilamellar appearance under transmission electron microscope. Seven fractions at 40, 50, 60, 110, 130, 140 and 150 kDa were identified as the antigenic membrane proteins in EITB with anti HY-1 rabbit serum. The common band at 60 kDa was detected both in antigenic fractions of plasma membrane and surface protein labelled with NHS-biotin. This result indicates that this protein is considered as a major surface antigen of T. vaginalis. The role of this surface antigen at 60 kDa should be studied further.

Animals↗

Changes of IgE production, splenic helper and suppressor T lymphocytes in mice infected with Paragonimus westermani.

Effects of Paragonims westermani infection were observed in mice on the change of serum IgE level, the number of peripheral eosinophils and the distribution of Thy 1.2+ (CD3), L3T4+ (CD4), and Lyt-2+ (CD8) splenic T lymphocytes without mitogen stimulation. BALB/c mice were infected with 20 metacercariae of P. westermani each. Total serum IgE increased at 3 weeks after the infection and reached a peak on week 4 and maintained high levels of IgE until the 23rd week. Peripheral eosinophil numbers increased at the second week and attained peak level on week 9. The frequency of L3T4+ (CD4) and Lyt-2+ (CD8) T lymphocytes decreased slightly until 4 weeks after the infection, but not significantly. Absolute number of L3T4+ and Lyt-2+ T lymphocytes, and the ratio of L3T4/Lyt-2 were not markedly changed over the period of observation. The frequency of Thy 1.2+ (CD3) T lymphocytes in the infected group slightly decreased until 4 weeks after the infection and showed significant reductions at the 2nd and 4th week of the infection (p < 0.05).

Animals↗