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Biomedical subjects

S H Oh

Publications and source records attributed to S H Oh.

At least 73 records · Page 4Linked to original sources

Prokaryotic expression and characterization of human AP DNA endonuclease.

The expression of major human apurinic/apyrimidinic DNA endonuclease (APEX) from its cDNA in E. coli (DH5 alpha) was attempted in order to obtain a biologically active recombinant APEX. E. coli cells were transformed by a prokaryotic translation vector (pGEX-4T-3) harboring APEX cDNA. GST-APEX fusion protein with a molecular weight of 6.3 KDa was induced by IPTG (1.0 mM) treatment. Western blot immunodetection identified the induced protein as the GST-APEX fusion protein. The survival rate of E. coli cells (DH5 alpha) transformed with pGEX-4T-3-APEX increased when the cells were treated with N-diethyl-N-nitrosamine (DENA) or 3'-methyl-4-monomethylaminoazobenzene (3'-MeMAB), indicating that APEX expression had a protective effect on the cytotoxicity of these carcinogens. The fusion protein extracted from E. coli cells and purified by GSH-agarose gel affinity chromatography exhibited APEX activity. Treatment of thrombin to the GST-APEX fusion protein and affinity purification followed by Sephacryl S-100 gel filtration resulted in APEX peptide with MW 36 KDa, which exhibited AP DNA repair activity (8,7000 EU/mg protein). N-ethylmaleimide (0.1 mM) or AMP (0.98 mM) inhibited APEX activity by 50% and kinetic analysis indicated that the recombinant APEX (rAPEX) had a Km value of 0.022 microM (AP sites for AP DNA) and the Ki value was 0.48 mM for AMP. These results indicated that E. coli cells expressing biologically active GST-APEX were resistant to the cell damage caused by chemical carcinogens and that rAPEX purified from E. coli cells transformed with APEX cDNA-inserted translation vector was similar to native APEX in some properties.

Carbon-Oxygen Lyases↗

Simultaneous reconstruction of the Achilles tendon and soft-tissue defect using only a latissimus dorsi muscle free flap.

The combined loss of the Achilles tendon and the overlying soft tissue in the young ambulant patient with expectations of a normal life is a challenging problem. These patients need not only soft tissue but also dynamic and functional reconstruction. Four cases of major defects of the Achilles tendon and overlying soft tissue after trauma are presented. In each case, the tendon and the overlying soft tissues were reconstructed using only a latissimus dorsi muscle free flap and overlying split-thickness skin graft. In conventional methods, evolved in the reconstruction of the Achilles tendon and soft tissue, the size of the defect was a limit. However, this technique can be used to reconstruct an extensive defect, including distal calf muscle to the plantar metatarsal area. In one case, the flap was harvested in a myocutaneous unit, and the skin portion was deepithelialized for the coverage and enough padding on the bony exposure area in reverse position. The purpose of the present study was to reevaluate the potential of denervated muscle flap for a force-bearing conduit as an alternative reconstructive method of the Achilles tendon. The denervated latissimus dorsi muscle in this study eventually experienced the process of atrophy and fibrosis but maintained its original length. Although there remained some atrophic muscle fibers, a fibrosis of the muscle fibers formed a tendon-like fibrous band, and so the action of the posterior calf muscle could be transmitted through the tendon-like fibrotic change of the denervated latissimus dorsi muscle. The advantages of this technique are that (1) it is a single procedure, (2) it is adaptable to a wide range of defect sizes, (3) it allows faster wound healing supported by well-vascularized tissues, (4) it produces satisfactory function of the ankle joint and a padding effect, and (5) it produces good contour of the posterior calf to the sole and an acceptable donor-site morbidity.

Achilles Tendon↗

Cloning and characterization of a tobacco cDNA encoding calcium/calmodulin-dependent glutamate decarboxylase.

In plants, gamma-aminobutyric acid (GABA), a major transmitter in the central nervous system in animals, is synthesized by glutamate decarboxylase (GAD), the activity of which is tightly modulated by Ca2+/calmodulin. To study the molecular mechanism of GAD regulation and examine the physiological role of GABA in plants, we isolated and characterized a 1771 bp tobacco cDNA clone, pNtGAD2. The 496 amino acid sequence deduced from pNtGAD2 showed 97, 92, and 81% identity to NtGAD1, petunia, and tomato GAD, respectively. The 26 amino acid sequence within the putative calmodulin binding domain at the carboxy terminus showed a typical alpha-helical structure with hydrophobic and charged amino acid clusters. The pNtGAD2-encoded 56 kDa protein interacted strongly with a monoclonal antibody against the petunia GAD and its GAD activity was stimulated markedly by the addition of exogenous calcium and calmodulin. The molecular sequence of pNtGAD2 and biochemical characteristics of the pNtGAD2-encoded protein confirmed that pNtGAD2 is a clone encoding a functional calmodulin-binding and Ca2+/calmodulin-dependent tobacco glutamate decarboxylase. This result indicates that tobacco plants also have Ca2+/calmodulin-dependent GADs.

Amino Acid Sequence↗

Rat oocytes fertilized in modified rat 1-cell embryo culture medium containing a high sodium chloride concentration and bovine serum albumin maintain developmental ability to the blastocyst stage.

A suitable chemically defined culture medium for 1-cell rat embryos (mR1ECM) was modified to obtain sperm penetration, and the developmental competence of oocytes fertilized in the medium was compared to that of oocytes fertilized in a traditional medium, modified Krebs-Ringer bicarbonate medium (mKRB). Sperm penetration was not observed when polyvinyl alcohol was replaced with BSA in mR1ECM (mR1ECM-BSA); the incidence was improved only when the osmolarity in mR1ECM-BSA was increased to that in mKRB (310 mOsm) by addition of NaCl. The proportion of oocytes penetrated in mR1ECM-BSA with NaCl increased (71.6 +/- 6.9%), which was not different compared to that in mKRB (76.7 +/- 13.7%). High incidences of sperm penetration (88.8 +/- 4.1% to 93.1 +/- 5.1%) were also observed when NaCl concentration in mR1ECM-BSA was increased from 76.7 mM to 100-130 mM. The incidence of embryos developing to the morula and blastocyst stages was higher when fertilized in mR1ECM-BSA containing 110-130 mM NaCl (91-94%) than in mKRB (70%). A total of 5 offspring were obtained after transfer of the morulae and blastocysts (69 embryos/7 females). These results demonstrate that a high developmental ability of rat embryos to the blastocyst stage is attained when the embryos have been fertilized in mR1ECM-BSA containing 110-130 mM NaCl and then cultured in mR1ECM.

Animals↗

Immunohistochemical study of 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase in the rat cardiovascular system.

The enzyme complex 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. It is known that the enzymatic activity of 3beta-HSD is present not only in classical steroidogenic tissues, but also in many peripheral tissues including cardiac tissue. To determine whether 3beta-HSD is present in rat non-steroidogenic tissues, we examined cardiovascular tissues including the ventricle, atrium, aortic arch, abdominal aorta, and inferior vena cava by immunohistochemistry and Western blotting using polyclonal antibody raised against a synthetic peptide of human placental 3beta-HSD. By Western blotting, protein bands immunoreactive for anti-3beta-HSD were detected at molecular weights of 42 and 37 kDa in both the ventricle and atrium, whereas only a 37 kDa band was recognized in both the aortic arch and abdominal aorta. By immunohistochemistry, immunoreactivity for 3beta,-HSD was detected in both the ventricular and atrial cardiocytes, while immunostaining was also found, though faintly, in the smooth muscles of the aortic arch, abdominal aorta, and inferior vena cava. These results suggest that cardiocytes may synthesize the steroidogenic 3beta,-HSD enzyme.

Animals↗

The causative organisms of bacterial meningitis in Korean children, 1986-1995.

Bacterial meningitis remains a serious cause of morbidity and mortality in childhood. Epidemiologic investigations have shown variability in disease risks among different populations and races. In Korea, however, basic epidemiologic information on bacterial meningitis in children is limited. The main purpose of this study was to analyze bacteriologically proven meningitis cases in terms of the relative frequency of causative organisms, mortality rate, and age distribution beyond the neonatal period. Data was obtained from the hospital records who had been diagnosed with bacterial meningitis at 13 general or university hospitals from 1986 through 1995. The patients had at least one positive CSF culture for bacteria. Of 140 cases of CSF culture-proven bacterial meningitis, 46.4% was < or =1 year, 62.1% was < or =2 years, 81.4% was < or =5 years cumulatively. Streptococcus pneumoniae was the most common bacteria responsible for 48 (35.0%) of all cases regardless of age, followed by Haemophilus influenzae for 48 (34.3%) and Neisseria meningitidis for 8 (6.4%) patients. The case fatality rate was 20.0%, 17.1%, and 16.7% for N. meningitidis, S. pneumoniae, and H. influenzae, respectively. In conclusion, the most common organisms of culture-proven bacterial meningitis in the last 10 years have been S. pneumoniae, H. influenzae, and N. meningitidis in order of frequency. Further study should be extended to nation-wide epidemiologic evaluation to show the incidence of bacterial meningitis caused by these three important organisms.

Adolescent↗

Ultrastructural changes of the internal elastic lamina in experimental hypercholesterolemic porcine coronary arteries.

The internal elastic lamina (IEL) serves as a barrier for cells and macromolecules between the intima and media in the vascular wall. We evaluated the morphological changes and quantitative assessments of the IEL architecture in the coronary circulation of pigs fed with a high cholesterol diet. Transmission electron microscopy (TEM) analysis of the IEL from hypercholesterolemic coronary arteries revealed fragmentation of the IEL associated with a decrease in the thickness. Confocal microscopy and scanning electron microscopy (SEM) revealed an altered pattern characterized by a large oval fenestration in the IEL of hypercholesterolemic vessels. Morphometric analysis of confocal microscopy images demonstrated that the IEL of cholesterol-fed animals were characterized by an increase in the minor diameter of the fenestrae (2.16+/-0.04 microm vs 3.32+/-0.06 microm, p=0.003) and a decrease in the fenestrae density (22,333+/-1,334/mm2 vs 17,552+/-931/mm2, p=0.015) compared to controls. The percentage of the IEL area covered by the fenestrae correlated with the intimal thickness (r=0.79, p=0.004). The immunoreactivity for matrix metalloproteinase-3 (MMP-3) increased in cholesterol-fed coronary arteries, predominantly in the neointima. This study demonstrates experimental hypercholesterolemia induced ultrastructural changes of the IEL in the coronary circulation. The IEL may play an important role in the development of structural changes which characterize the early phase of coronary atherosclerosis.

Animals↗

Experimental hypercholesterolemia induces ultrastructural changes in the elastic laminae of rabbit aortic valve.

Atherosclerosis is the most severe problem in the high-pressure systemic circulation and similar changes also occur in the high-pressure loading valve. This study was designed to test the hypothesis that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. However, it is not known whether this process is also taking place in the cardiac valve at the early stage of atherosclerosis. Animals were fed either a high cholesterol diet (n = 5) or a control diet (n = 5) for 10-12 weeks. Histologic analysis demonstrated that subendothelial thickening and foam-cell infiltration were evident in the arterialis of aortic valves. Confocal microscopy revealed an altered pattern characterized by fragmentation and disorganization of the arterialis elastic laminae of hypercholesterolemic valves. Computerized digital analysis of the images obtained by confocal scanning microscopy demonstrated that compared to normal valves, the arterialis elastic laminae of hypercholesterolemic valves decreased in percentage of their elastin content (29.03 +/- 1.10% vs. 42.94 +/- 1.35%, p = 0.023). Immunohistochemical staining for matrix metalloproteinase-3 (MMP-3) revealed MMP-3 immunoreactivity was increased in hypercholesterolemic valves, predominantly in the arterialis. This study demonstrated that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. The arterialis endothelium of the aortic valve may be a more atherosclerosis-prone area compared with the ventricularis. The presence of ultrastructural defect in the elastic laminae may play a role in chronic degenerative change and a resultant valvular dysfunction.

Animals↗

Multiple cerebellar infarction due to vertebral artery obstruction and bulbar symptoms associated with vertical subluxation and atlanto-occipital subluxation in ankylosing spondylitis.

Ankylosing spondylitis (AS) results in disease-specific inflammation at the site of ligamentous insertion into the bone. Atlantoaxial joint subluxation and vertical subluxation of the axis may occur as a consequence of instability resulting from the inflammatory process. Spontaneous anterior atlantoaxial subluxation is a well recognized complication in about 2% of patients with AS, and presents with or without signs of spinal cord compression. Vertical subluxation may follow anterior or posterior subluxation. It was noted in 3-8% of patients with rheumatoid arthritis, but is an exceedingly rare complication of AS. Moreover, it has never been reported that multiple cerebellar infarction and bulbar symptoms developed spontaneously due to atlanto-occipital subluxation and vertical subluxation in a patient with a long [corrected] history of AS. We describe a man with AS who developed multiple cerebellar infarction due to vertebral artery obstruction and bulbar symptoms associated with atlanto-occipital subluxation and vertical subluxation.

Arterial Occlusive Diseases↗

Transgenic tobacco expressing a foreign calmodulin gene shows an enhanced production of active oxygen species.

A strategy for elucidating specific molecular targets of calcium and calmodulin in plant defense responses has been developed. We have used a dominant-acting calmodulin mutant (VU-3, Lys to Arg115) to investigate the oxidative burst and nicotinamide co-enzyme fluxes after various stimuli (cellulase, harpin, incompatible bacteria, osmotic and mechanical) that elicit plant defense responses in transgenic tobacco cell cultures. VU-3 calmodulin differs from endogenous plant calmodulin in that it cannot be methylated post-translationally, and as a result it hyperactivates calmodulin-dependent NAD kinase. Cells expressing VU-3 calmodulin exhibited a stronger active oxygen burst that occurred more rapidly than in normal control cells challenged with the same stimuli. Increases in NADPH level were also greater in VU-3 cells and coincided both in timing and magnitude with development of the active oxygen species (AOS) burst. These data show that calmodulin is a target of calcium fluxes in response to elicitor or environmental stress, and provide the first evidence that plant NAD kinase may be a downstream target which potentiates AOS production by altering NAD(H)/NADP(H) homeostasis.

Calcium↗

Effect of mandibular setback surgery on occlusal force.

PURPOSE: This study investigated the effect of mandibular setback surgery on occlusal force and evaluated the extent to which postsurgical changes in such force can be explained by the type of operation, the duration of maxillo-mandibular fixation (MMF), and the changes in the mechanical advantage of the jaw musculature. MATERIALS AND METHODS: Maximal molar bite force was measured before surgery and at MMF removal, and 3, 6, and 12 months thereafter in 26 patients with mandibular prognothism. To correlate bite force and skeletal change, the cephalometric tracings were measured, tabulated, and statistically analyzed. RESULTS: Mean bite force was 13.7 kg before surgery, 7.6 kg at MMF removal, 14.2 kg at 3 months, 19.7 kg at 6 months, and 26.1 kg at 12 months post-surgery. The bite force was positively correlated with the surgical change in mandibular plane angle and mandibular body length. The recovery of bite force was significantly affected by the type of operation and duration of MMF. CONCLUSIONS: To hasten recovery and increase bite force after orthognathic surgery, long periods of MMF and injury to the masticatory muscles should be avoided.

Adolescent↗

Denaturation of circular or linear DNA facilitates targeted integrative transformation of Streptomyces coelicolor A3(2): possible relevance to other organisms.

Using Streptomyces coelicolor A3(2) protoplasts, the number of transformants obtained by homologous recombination of incoming double-stranded circular DNA with the recipient chromosome was greatly stimulated by simple denaturation of the donor DNA. This procedure was very effective with inserts over a ca. 100-fold size range, the largest tested being ca. 40-kb inserts in cosmids. These observations led to transformation experiments with linearized cloned DNA and randomly sheared genomic DNA. In both cases, DNA denaturation led to significant levels of transformation. Most of the transformants had resulted from the predicted homologous recombination events. A number of genetic manipulations will be made easier or possible by these procedures.

Bacterial Proteins↗

Increase in expression of IL-4 and IL-5 mRNA in the nasal mucosa of patients with perennial allergic rhinitis during natural allergen exposure.

To understand the role of cytokines in allergic nasal disease in humans, we designed this study to investigate the expression of cytokine mRNA in the nasal mucosa of 10 control subjects and 10 patients with perennial allergic rhinitis using a reverse transcription-polymerase chain reaction (RT-PCR) for IL-4, IL-5 (Th2 cytokines), and IFN-gamma (Th1 cytokine). Both IL-4 and IL-5 mRNAs were expressed in a significantly larger number of patients with allergic rhinitis than in control subjects. When the levels of expression of cytokines and beta-actin mRNA were quantitated, the mean density ratios (cytokine/beta-actin) for IL-4 and IL-5 were higher in patients with allergic rhinitis than in control subjects. In contrast, there were no differences in the number of subjects who showed expression of IFN-gamma mRNA, or in mean density ratios for IFN-gamma between the two groups. This study suggests that both IL-4 and IL-5 mRNAs are expressed almost universally in the nasal mucosa of patients with perennial allergic rhinitis during natural allergen exposure and also that Th2-type cytokines play a crucial role in the development of allergic nasal diseases in vivo.

Adult↗

EBV-elicited familial hemophagocytic lymphohistiocytosis.

Familial hemophagocytic lymphohistiocytosis (FHL) is a rapidly fatal illness, usually encountered in infancy, characterized by fever, hepatosplenomegaly, pancytopenia, and central nervous system involvement. Microscopic examination of tissue shows a non-malignant lymphohistiocytic infiltrate, with prominent erythrophagocytosis. FHL is an autosomal recessive hereditary disorder but may develop secondarily to other conditions such as immunosuppression, malignancies, fat overload and certain infections. We recently experienced a case of siblings developing FHL, which may be associated with EBV infection.

Child, Preschool↗

Sensory organ generation in the chick inner ear.

There are a total of eight sensory organs in the chick inner ear. Each sensory organ has a distinct structure tailored for its function, and its morphology is well characterized. However, the origin of these sensory organs and the lineage relationships among them are largely unknown. In this report, we show that BMP4 (bone morphogenetic protein), a secreted protein of the TGF-beta gene family, is the earliest sensory marker identified to date for the chick inner ear. In addition to BMP4, we show that Msx-1 is a sensory marker for the three cristae, the lagena, and macula neglecta. P75NGFR (nerve growth factor receptor) is a marker for the three cristae only. Based on the expression pattern of these three genes-BMP4, Msx-1, and p75NGFR-it is estimated that the first sensory organs to be generated were the superior and posterior cristae at stage 19, followed by the macula sacculi at stage 20, the lateral crista at stage 22, the basilar papilla and lagena at stage 23, the macula utriculi at stage 24, and the macula neglecta at stage 29. The age of generation of each sensory organ as defined by the first appearance of these molecular markers is well in advance of the histological differentiation. In addition, the differential gene expressions in each presumptive sensory organ may contribute to the distinct structure of the mature organ.

Animals↗

Differential expression of bone morphogenetic proteins in the developing vestibular and auditory sensory organs.

The genes responsible for the formation of various sensory organs in the inner ear are not known. There are eight sensory organs in the chick inner ear, and our previous study showed that all presumptive sensory organs initially express bone morphogenetic protein 4 (BMP4), a member of the transforming growth factor (TGF)-beta gene family. To address the potential role of BMPs in the patterning of different sensory organ structures, we investigated the expression of BMP4, BMP5, and BMP7 during sensory organ differentiation in the chick inner ear. The gene expression pattern of BMP5, although similar to that of BMP4, was transient and disappeared by embryonic day 3.5 (E3.5). In contrast, BMP7 gene expression was quite extensive, starting in the otic placode. By E5, gene expression patterns of BMP4 and BMP7 differed among vestibular and auditory sensory organs. In the vestibular sensory organs, BMP7 gene expression segregated from the main sensory tissue areas at the onset of differentiation, whereas BMP4 expression concentrated in supporting cells. In the cochlea, however, BMP7 gene expression became restricted to sensory tissue over time and eventually concentrated in supporting cells, whereas BMP4 gene expression was localized to hair cells. The different BMP expression patterns in developing auditory and vestibular sensory organs may help to shape each respective sensory structure. Furthermore, the expression of BMP4 in the cochlea also revealed an interesting pattern of sensory cell differentiation: the distal portion of the cochlea differentiates first, and the tall hair cells develop before the short hair cells.

Amino Acid Sequence↗