In this study, we are to produce the single chain variable fragment (scFv) antibodies against surface protein of hepatitis B virus (HBV) using antibody phage display technique. Balb/c mice were immunized with preS1 and cDNAs of heavy and light chains of splenic B cells from immunized mice were prepared using RT-PCR. Two cDNAs were linked with (64S) linker DNA under recombination PCR to produce single chain Fv DNA. After digestion of scFv DNA with Sp 1 and Not 1, the digested DNA was ligated into pCANTAB 5E and electroporated into E. coli XL1-Blue to prepare scFv-library. The size of library was 1 * 10' pfu/ml. Phage antibodies (phabs) against preS1 were rescued with M13K07 helper phages, and preS1-binders were selected through 3 times of panning using 96 well microtitre plates. Phage antibody clones were assayed directly for the ability to bind preS1 by ELISA. And then 7 phage antibody clones had high ELISA signals against preS1. Phabs from preS1-specific pMsc-17 had the strongest ELISA signal to preS1. Phabs from pMsc-17 were used for Western blot to preS1 and the results revealed that it was specific to preS1. To prepare the soluble scFv antibody, phabs from pMsc-17 were transfected into non-suppressor E. coli HB2151, and grown under 1 mM IPTG. Soluble scFv antibody was mainly accumulated in the periplasmic space, but small amount of antibody was secreted into culture media.
Animals ; Antibodies ; B-Lymphocytes ; Bacteriophages* ; Blotting, Western ; Cell Surface Display Techniques ; Clone Cells ; Culture Media ; Digestion ; DNA ; DNA, Complementary ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Isopropyl Thiogalactoside ; Mice* ; Periplasm ; Polymerase Chain Reaction ; Recombination, Genetic ; Single-Chain Antibodies*

No abstract available.
Antibodies* ; Chungcheongnam-do* ; Diagnosis* ; Immunoglobulin G* ; Immunoglobulin M* ; Orientia tsutsugamushi* ; Rickettsia* ; Scrub Typhus*

No abstract available.
Antibodies* ; Chungcheongnam-do* ; Diagnosis* ; Immunoglobulin G* ; Immunoglobulin M* ; Orientia tsutsugamushi* ; Rickettsia* ; Scrub Typhus*

BACKGROUND: The potential adverse effects of ketamine in neurosurgical anesthesia have been well established. However, the effects of ketamine on intracranial pressure (ICP) and hemodynamics during general anesthesia remain unclear. The purpose of this study was to assess the effects of ketamine on hemodynamics and ICP in anesthetized, ventilated rabbits. METHODS: Thirty rabbits were divided into three groups: Group 1 (n=10) received 1 ml/kg normal saline iv; Group 2 (n=10) received 0.5 mg/kg ketamine iv; Group 3 (n=10) received 1.0 mg/kg ketamine iv. After induction with thiopental, anesthesia was maintained with isoflurane and nitrous oxide in oxygen. During controlled ventilation, ICP, mean arterial pressure (MAP), cerebral perfusion pressure (CPP) and heart rate (HR) were measured. The ICP was measured using Ladd ICP monitoring system. All variables were evaluated at baseline and for 30 min following ketamine. RESULTS: In group 1, ICP, MAP, CPP and HR were unchanged over the course of the study. In group 2, ICP, MAP and CPP were unchanged. HR increased at 1, 3 and 5 min (p<0.01), 10 and 20 min (p<0.05) after injection. In group 3, ICP, MAP and CPP increased at 1 and 3 min (p<0.01) after injection. HR increased at 1, 3 and 10 min (p<0.01), 5 min (p<0.05) after injection. CONCLUSIONS: These results suggest that 0.5 and 1.0 mg/kg of ketamine don't significantly affect the hemodynamics and ICP in anesthetized, mechanically ventilated rabbits.
Anesthesia* ; Anesthesia, General ; Arterial Pressure ; Heart Rate ; Hemodynamics* ; Intracranial Pressure* ; Isoflurane ; Ketamine* ; Nitrous Oxide ; Oxygen ; Perfusion ; Rabbits* ; Thiopental ; Ventilation

Human monoclonal antibodies have considerable potential in the prophylaxis and treatment of viral disease. By cloning human Ig gene segments from the B cells of volunteer into pComb3 phagemid vector, antibody library was created of filamentous phage particles displaying Fab fragments on their surface after being rescued with M13KO7 helper phages. The size of library was 7x10' pfu. Phage antibodies (phabs) were panned against biotinylated preS1 using streptavidine coated Dynabead. The soluble Fab antibodies were prepared from phagemid colonies and assayed directly for the ability to bind preS1 by ELISA. And then 3DW and SGW specific to preS1 which have both heavy and light chain to form Fab fragment, were selected. The soluble Fab antibody from 3DW was expressed highly at the concentration of 0.1 - 1.0 mM of IPTG, and 5 hours postinduction. The soluble antibodies from 3DW and SGW showed their relative affinities of 2x10' M ', and Sx10 M ', respectively, and the specificities to preS1 on ELISA. Our results suggest that antibody phage display library is very useful method to generate the human monoclonal antibody and that the human Fab monoclonal antibodies specific to preS1 selected in this study open the way to treat hepatitis B as a component of passive irnmunotherapeutics.
Antibodies ; Antibodies, Monoclonal ; B-Lymphocytes ; Bacteriophages* ; Clone Cells ; Cloning, Organism ; Enzyme-Linked Immunosorbent Assay ; Genes, Immunoglobulin ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Humans* ; Immunoglobulin Fab Fragments ; Isopropyl Thiogalactoside ; Streptavidin ; Virus Diseases ; Volunteers

Secondary osteoarthritis of hip joints is a common disease and is frequently followed by chronic sublexation and dislocation. Twenty four case of the secondary osteoarthritis associated with chronic subluxation and dislocation of the hip joints were evaluated with plain radiography and computed tomography. We retrospectively analyzed 1) the ossification and calcification of the acetabular labrum, 2) the thickeness of the quadrilateral plate of the ilium, and 3) anteroposterior diameter of the acetabulum. The changes of the hip joints in subluxation (n=14) revealed ossification of the acetabular labrum in 12 cases (86%), thickening of the quadrilateral plate of the ilium in 11 cases (78%) but anteroposterior diameter of the acetabulum was not changed. The changes of the hip joints in dislocation (n=10) revealed no evidence of the ossification of the acetabular labrum, thickening of the quadrilateral plate of the ilium in 10 cases (100%) and decreased anteroposterior diameter of the acetabulum. We conclude that CT findings of subluxation and dislocation of the hip joints can be helpful in the evaluation of the secondary osteoarthritis of the hip joints.
Acetabulum ; Dislocations* ; Hip Joint* ; Hip* ; Ilium ; Joints ; Osteoarthritis ; Osteoarthritis, Hip ; Radiography ; Retrospective Studies

Dental restorative materials must have the physical properties to withstand wear and corrosion. Base metal alloys possess better mechanical properties and lower price than the gold alloys. For these reason such alloys have largely replaced the precious metal alloys. One aspect to consider is the release of metal substances to oral environment. The release of elements from dental alloys is a continuing concern because the elements may have the potentially harmful biological effects on local tissue. The purpose of this study was to minimize metal release on the nonprecious metal surfaces by ion bea assisted deposition(IBAD) of titanium nitride (TiN). Ni-Cr-Be alloys with and without TiN coatings were secured in an wear test machine opposing ruby ball to determine their relative resistance to wear with 100m, 200m, 300m and 400m sliding distance. And the corrosion behavior of the Ni-Cr-Be alloys with and without TiN coating and 3 dental noble alloys have been studied. Potentiodynamic curves were used to analyse the corrosion characteristics of the alloys. The measurement of the released Ni and Ci ions was conducted by analysis of the electrolyte solution with atomic absorption spectroscopy. The results were as follows : 1. The critical sliding distance that wore down TiN coating of 2.5micrometer thickness in this study condition was 300m. 2. Ion beam assisted deposition of TiN showed a good surface modification with respect to the properties of wear and corrosion resistance. 3. X-ray diffraction showed that the strongest peak of TiN(111) in the coatings. 4. The release of Ni and Cr ions from alloys measured by means of atomic absorption spectroscopy was reduced by ion beam assisted deposition of TiN.
Absorption ; Alloys* ; Corrosion ; Dental Alloys ; Gold Alloys ; Ions ; Spectrum Analysis ; Tin* ; Titanium ; X-Ray Diffraction

No abstract available.
Seizures* ; Sparganosis*

PURPOSE: MR findings of the spinal metastasis and the tuberculosis are well known, but sometimes it might be difficult to differentiate these lesions. Therefore we reviewed and analyzed the MR findings which would be useful for the differentiation. MATERIALS AND METHODS: T1- and T2-weighted spin echo images and gadolinium-enhanced Tl-weighted images were obtained with 1. 5T and 1. 0T superconductive MR imager. We reviewed MR findings in 16 cases of spinal metastases and 24 cases of spinal tuberculosis in terms of signal intensity, contrast enhancement pattern, disc space involvement, spinal canal compressing feature and paraspinal soft tissue mass. RESULTS: The signal intensities of both lesions were hypointense on T1WI and hyperintense on T2WI except those of the metastatic lesions from the prostatic carcinoma. Heterogeneous enhancement was noted in 63% of metastasis, whereas peripheral rim enhancement was noted in 83% of spinal tuberculosis(p (.001). Spinal canal compression by collapsed vertebra was only noted in spinal metastasis, and that by paraspinal soft tissue was noted in both spinal metastasis and tuberculosis(p (.001). Disc space invasion was noted in 19% of spinal metastasis and 88% of spinal tuberculosis. Spinal tuberculosis was common at lower thoracic spine(T10) and typically involved two or more adjacent vertebral bodies(96%). CONCLUSION: The important differential point between spinal metastasis and tuberculosis was the enhancement pattern, involvement of two or more contiguous vertebral bodies and the feature of spinal canal compressing. The secondary importance was the disc space involvement pattern.
Neoplasm Metastasis* ; Spinal Canal ; Spine ; Tuberculosis ; Tuberculosis, Spinal*

BACKGROUND: Hereditary spherocytosis(HS) is a clinically and biochemically very heterogeneous disorder The purpose of this study is to detect erythrocyte membrane protein abnormalities by SDS-PAGE and to investigate the frequency of erythrocyte membrane protein defects in hereditary spherocytosis and correlation between some of the hereditary spherocytosis biochemical subsets and the selected clinical phenotype. METHODS: We evaluated the clinical and laboratory characteristics of 14 normal healthy persons and 23 hereditary spherocytosis patients and 8 their family members. The patients were divided into three groups based on clinical and hematological severity(mild, typical, severe). In addition to routine hematologic determlnatlons, osmotic fragility and autohemolysis, RBC membrane protein analysis were performed in all patients by densitometric tracing of SDS-PAGE(sodium dodecyl sulphate polyacrylamide gel electrophoresis) stained by Coomassle blue utilizing both the discontinuous buffer system of Laemmli with acrylamide linear gradient from 4% to 12% and the continuous buffer system of Fairbank with exponential gradient of acrylamide from 3.5% to 17%. RESULTS: 1) The patients could be seperated into three classes of different clinical severity as mild(3 cases), moderate(16 cases) and severe(4 cases) on the clinical feature. 2) Eighteen patients(82.6%) among 23 hereditary spherocytosis revealed abnormal erythrocyte membrane protein and we detected six patients(26.1%) with spectrin deficiency combined with ankyrin reduction, 4 patients(17.4%) with ankyrin deficiency, 4 patients(17.4%) with isolated spectrin deficiency and 3 patients(13.0%) with band 3 deficiency. Five HS patients(21.7%) showed normal RBC membrane protein. 3) Eight HS and their family members showed same RBC membrane protein deficiency. 4) The type and degree of RBC membrane protein reduction were variale with spectrin at 66~94%, with ankyrin at 48~82% of normal levels. These showed that each patient had different clinical severities according to different RBC membrane protein levels and type. CONCLUSION: RBC membrane protein abnormalities were observed in 82.6% of HS patients. The combined spectrin and ankyrin deficiency is the most common molecular defect in HS. The clinical severity and biochemical expression is heterogeneous. SDS-PAGE analysis of RBC membrane protein was provided the diagnosis of RBC membrane defects and basic molecular studies. We believed that the early identification of the biochemical defect responsible for HS is important because it is helpful starling point for the identification of the primary molecular defect, and it could help to anticipate the clinical outcome of the disease. For these reasons, we consider the SDS-PAGE of the red cell membrane to be of crucial importance for a complete evaluation of children with HS. Further studies with more cases would be to clarify the correlation between clinical and biochemical phenotypes.
Acrylamide ; Ankyrins ; Cell Membrane ; Child ; Diagnosis ; Electrophoresis, Polyacrylamide Gel* ; Erythrocyte Membrane* ; Erythrocytes* ; Erythrocytes, Abnormal ; Humans ; Membrane Proteins ; Membranes ; Osmotic Fragility ; Phenotype ; Population Characteristics* ; Spectrin ; Starlings