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*

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

Santonin-kainic acid complex was evaluated as a chemotherapeutic of the mass treatment of the Ascaris lumbricoides infection in Korea. The results could be summarized as follows: The negative conversion rate was 82.9% in average in 4 treated groups. Some variations of negative conversion rate among the treated groups were noticed. The egg reduciton rate was 97.7% in average and the results were rather uniform among the three evaluated groups. By the analysis of egg reduction, it seems that the lightly infected cases whose E.P.G. were under 5,000 were resistant to treatment with the less reduced egg output. After the treatment with this complex, the number of egg discharged cases were reduced to 1.84% and the average number of discharge eggs per incompletely treated or untreated cases were reduced to 12.5% compared with the level of before-treatment egg output. The successive observations of the pattern of worm expulsion after drug intake was made. The worms were mostly expelled in the stool within 3 days, and 58.9% of total expelled worms were collected within 24-hour stool. The minimum length of the immature worms expelled was 7.6cm. Among the 659 Ascaris collected in the first-day stool from 91 rural people, 8.5% were in the range of 7.6-12.2 cm-long, immature worms. The sex ratio, male: female= 0.69: 1.
parasitology-helminth-nematoda ; Ascaris lumbricoides ; chemotherapy ; Santonin ; kainic acid

No abstract available.
Antigen-Antibody Complex* ; Syphilis*

No abstract available.
Heart Septal Defects, Atrial* ; Hemodynamics*

Various neurotransmitters have been proposed as possible mediators of penile erection. Especially, norepinephrine and serotonin might have a important role in sexual arousal and penile erection. And it could be hypothesized that the psychogenic impotence is associated with the depletion or imbalance of norepinephrine and serotonin from evidences such as the symptomatic manifestation of depression and the antidepressant-induced sexual dysfunction. The authors investigates the association of norepienphrine and serotonin with psychogenic impotence. The psychogenic impotent group(PIG) consisted of twenty-three patients with psychogenic impotence and the controlled group(CG) consisted of twenty-seven patients without psychogenic impotence. PIG had no organic cause accounting for their erectile dysfunction. The Beck Depression Inventory(BDI) and the State-Trait Anxiety Inventory(STAI) were applied to each subject to assess mood, state anxiety(SA) and trait anxiety(TA). Plasma norepinephrine level from systemic blood and 5-hydroxyindoleacetic acid(HIAA) levels from 24-hours urine were measured in each subject. The mean score of BDI of PIG was significantly higher than that of CG(p=0.015). PIG had a tendency of higher TA compared with CG(p=0.054). And also SA was higher in PIG, bud did not show significant difference(p=0.193). The level of norepinephrine was significantly lower in patient with psychogenic impotence(p=0.000). And the level of 24-hours urine 5-HIAA was lower in PIG but did not show significant difference(p=0.494). Although the authors did not exclude depressive disorders in PIG, the present findings suggest that psychogenic impotence might have higher depressive mood and trait anxiety, and be associated with the depletion of norepinephrine in systemic blood.
Anxiety ; Arousal ; Depression ; Depressive Disorder ; Erectile Dysfunction* ; Humans ; Hydroxyindoleacetic Acid ; Male ; Neurotransmitter Agents ; Norepinephrine* ; Penile Erection ; Plasma ; Serotonin*

Left atrial myxoma is well-known mimicker of systemic vasculitis. In general, however, these patients showed some symptoms and signs of cardiovascular disease. We experienced a patient with left atrial myxoma who first presented as vasculitis without any symptoms and signs suggesting cardiovascular disorders. A 45 year-old lady showed tender skin nodules, livedo reticuiaris-like skin lesions, severe constitutional symptoms, multiple cerebral infarctions, and elevated ESR and globulin. Skin biopsy findings were thought to be consistent with polyarteritis nodosa. Echocardiography was performed as a routine test for evaluating suspected vasculitis patients and it revealed left atrial myxoma. However, we were not able to detect any auscultation abnormality, even after echocardiography, After echocardiography, we found the tumor emboli in skin biopy specimen with deeper section. This case demonstrated the importance of suspecting the possibility of left atrial myxoma when performing diagnostic work-up for vasculitis.
Auscultation ; Biopsy ; Cardiovascular Diseases ; Cerebral Infarction ; Echocardiography ; Humans ; Middle Aged ; Myxoma ; Polyarteritis Nodosa ; Skin ; Systemic Vasculitis ; Vasculitis

BACKGROUND: Since the demonstration of the fact that vascular relaxation by acetylcholine(Ach) results from the release of relaxing factor from the endothelium, the identity and physiology of this endothelium-derived relaxing factor(EDRF) has been the target for many researches. EDRF has been identified as nitric oxide(NO). With the recent evidences that EDRF is an important mediator of vascular tone, there have been increasing interests in defining the role of the EDRF as a potential mediator of hypoxic pulmonary vasoconstriction. But the role of EDRF in modulating the pulmonary circulation is not compeletely clarified. To investigate the endotbelium-dependent pulmonary vasodilation and the role of EDRF during hypoxic pulmonary vasoconstriction, we studied the effects of N(G)-monornethyl-L-arginine(L-NMMA) and L-arginine on the precontracted pulmonary arterial rings of the rat in normoxia and hypoxia. METHODS: The pulmonary arteries of male Sprague Dawley(300~350g) were dissected free of surrounding tissue, and cut into rings. Rings were mounted over fine rigid wires, in organ chambers filled with 20ml of Krebs solution bubbled with 95 percent oxygen and 5 percent carbon dioxide and maintained at 37℃. Changes in isometric tension were recorded with a force transducer(FT. 03 Grass, Quincy, USA). RESULTS: 1) Precontraction of rat pulmonry artery with intact endothelium by phenylephrine(PE, 10(-6)M) was relaxed completely by acetylcholine(Ach, 10(-9) -10(-5)M) and sodium nitroprusside (SN, 10(-9) -10(-5)M), but relaxing response by Ach in rat pulmonary artery with denuded endothelium was significantly decreased. 2) L-NMMA(10-4M) pretreatment inhibited Ach(10(-9) -10(-5)M)-induced relaxation, but L-NMMA(10-4M) had no effect on relaxation induced by SN(10(-9) -10(-5)M). 3) Pretreatment of the L-arginine(10(-4)M) significantly reversed the inhibition of the Ach(10(-9) -10(-5)M)-induced relaxation caused by L-NMMA(10(-4)M). 4) Pulmonary arterial contraction by PE(10(-6)M) was stronger in hypoxia than normoxia but relaxing response by Ach(10(-9) -10(-5)M) was decreased. 5) With pretreatment of L-arginine(10(-4)M), pulmonary arterial relaxation by Ach(10(-9) -10(-5)M) in hypoxia was reversed to the level of relaxation in normoxia. CONCLUSION: It is concluded that rat pulmonary arterial relaxation by Ach is dependent on the intact endothelium and is largely mediated by NO. Acute hypoxic pulmonary vasoconstriction is related to the suppression on NO formation in the vascular endothelium.
Animals ; Anoxia ; Arginine ; Arteries ; Carbon Dioxide ; Endothelium ; Endothelium, Vascular ; Humans ; Male ; Nitric Oxide* ; Nitroprusside ; omega-N-Methylarginine ; Oxygen ; Physiology ; Poaceae ; Pulmonary Artery ; Pulmonary Circulation ; Rats* ; Relaxation* ; Vasoconstriction* ; Vasodilation

OBJECTIVE: To investigate the characteristics of the motor cortex map for abductor pollicis brevis muscle (APB) using transcranial magnetic stimulation (TMS) in normal subjects. METHOD: Ten adults without neurological disorder were studied. A piece of cloth which marked at 1 cm interval was fixed on the head of the subject. The motor cortex mapping for APB was done with butterfly magnetic stimulator, and then with round magnetic stimulator. RESULTS: The average optimal scalp position for left APB was located on lateral 6.2 cm, anterior 0.1 cm from Cz and that for right APB was located on lateral 6.0 cm, anterior 0.1 cm from Cz when stimulated with butterfly magnetic stimulator. The differences between hemispheres were less than 1 cm in the location of optimal scalp position and less than 10% in excitatory threshold (ET) irrespective of magnetic stimulator. The ipsilateral motor evoked potential (MEP) was not evoked in all subjects. The ET when stimulated with butterfly magnetic stimulator was higher to that when stimulated with round magnetic stimulator. CONCLUSION: We conclude that TMS using butterfly and round magnetic stimulator is useful for the motor cortex mapping.
Adult ; Butterflies ; Evoked Potentials, Motor ; Head ; Humans ; Motor Cortex* ; Nervous System Diseases ; Scalp ; Transcranial Magnetic Stimulation*

OBJECTIVE: To investigate the utilization of medical rehabilitation services and the degree of satisfaction about rehabilitation services in patients with brain disorders. METHOD: A total of 1903 patients agreed to participate in this study and were interviewed from September 2005 to May 2006. The subjects completed the questionnaires about the utilization of medical rehabilitation services and the degree of satisfaction with those treatments. Pearson's chi-square test, Student t-test and frequency analysis were used for statistical analysis. RESULTS: A 78.0 percent of patients received inpatient rehabilitation treatment. A 66.9 percent of all patients were served of only physical therapy and 31.6 percent received both physical and occupational therapy. The main reason why patients could not have a chance to experience rehabilitation treatment was associated with environmental problems, such as the ignorance of the need about rehabilitation treatment, or the insufficient communication between doctors and patients. Most patients (54.6%) were satisfied with the rehabilitation treatment. However, as the number of admission was increased, patients tended to be less satisfied with the rehabilitation treatment. CONCLUSION: The inpatient rehabilitation treatment was limitedly served to patients with brain disorders mostly when the patients required rehabilitation services. It is necessary to provide more effective and various rehabilitation services to patients under the comprehensive guideline of the process of rehabilitation services.
Brain ; Brain Diseases ; Humans ; Inpatients ; Korea ; Occupational Therapy ; Surveys and Questionnaires