The ability to introduce recombinant DNA molecules back into mycobacteria would greatly increase the potential of molecular genetic approaches for the study of mycobacteria as well as for the use in clinical purposes. We have initiated the construction of vectors that facilitates the introduction of recombinant DNA into mycobacteria. The vector was designed to contain replicons for multiplication in mycobacteria and Escherichia coli, a promoter for gene expression, a drug resistant gene for selecting transformants, and a few restriction enzyme sites for convenient cloning. Constructed Mycobacterium-E. coli shuttle vector named p YMC (hsp60) was shown to transform M. smegmatis at high efficiency and maintain plasmid at stable level. The ability of the vector to express cloned foreign gene was also monitored by measuring the expressed level of luciferase gene which was used as a reporter. High level of luciferase activity in M. smegmatis with pYMC (hsp60:luc) was detected confirming successful construction of Mycobacterium-E. coli shuttle vector.
Clone Cells ; Cloning, Organism ; DNA, Recombinant ; Escherichia coli* ; Escherichia* ; Gene Expression ; Genetic Vectors* ; Luciferases ; Molecular Biology ; Mycobacterium* ; Plasmids ; Replicon

The 38 kDa protein of Mycobacterium tuberculosis, which was known previously as antigen 5, has been extensively used in the serodiagnosis of tuberculosis. In an attempt to develop and evaluate a serodiagnostic test using the antigen, we expressed the 38 kDa protein in BCG and its seroreactivity was compared to that expressed in Escherichia coli. The coding region of the 38 kDa protein was amplified by PCR, and the gene was cloned into a Mycobacterium-E. coli shuttle expression vector pYMC-his and pQE30 expression vector and expressed in BCG and E. coli, respectively. Both recombinant 38 kDa proteins showed strong seroreactivity against pooled serum from tuberculosis patients. There was no significant difference in seroreactivity between the two recombinant antigens in sera from the far advanced tuberculosis patients. However, of 25 tuberculosis patients graded as ""minimal"" by chest X-ray, 5 (20.0%) were seropositive by r38 kDa expressed in E. coli, while 8 (32.0%) by that expressed in BCG. Likewise, higher seroreactivity by r38 kDa expressed in BCG was found in sera from the moderately advanced tuberculosis. This study thus indicates that the recombinant 38 kDa expressed in BCG is more effective than that expressed in E. coli in detecting antibodies to the native 38 kDa protein of M. tuberculosis in sera from minimally affected tuberculosis patients.
Antibodies ; Clinical Coding ; Clone Cells ; Escherichia coli ; Humans ; Mycobacterium bovis* ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymerase Chain Reaction ; Serologic Tests* ; Thorax ; Tuberculosis*

No Abstract Available.
Brucella abortus* ; Brucella* ; Immunity, Humoral* ; Jeju-do* ; Recombinant Proteins*

Direct surgical repair of complex intracranial vascular lesions is difficult. Sometimes the neurosurgery is performed under circulatory arrest, profound hypothermia and barbiturates cerebral protection. Total ischemia is tolerated for 30~60 minutes because oxygen requirements of the brain decrease exponentially as body temperature is lowered. We experienced that this technique was successfully used for inoperable basilar artery aneurysm. We reviewed the surgical and anesthetic considerations of basilar artery aneurysm.
Aneurysm* ; Barbiturates ; Body Temperature ; Brain ; Hypothermia* ; Intracranial Aneurysm ; Ischemia ; Neurosurgery ; Oxygen

As an effort to develop a rapid and sensitive diagnostic test, we produced previously a monoclonal antibody (MAb) specific to the lipoarabinomannan (LAM) antigen and used in a sandwich ELISA for detection of mycobacterial antigens in sputum. In this study, we attempted to improve the antigen detection assay by combination of af5nity-purified antibodies against Mycobacterium tuberculosis soluble antigen and anti-LAM MAb. With the new assay, the LAM antigen was detectable as low as 2 ng/ml, and none of 10 gram-negative and gram-positive organisms gave significant absorbance, thus indicating the specific detection of mycobacterial antigens. Sputum samples from 62 patients who were suspected having tuberculosis and from 37 healthy controls were examined. The sensitivity of the antigen detection assay ranged from 0% in the 1+ culture group to 78.8% in the 3+ culture group. When the results were combined, 15 of 24 culture-positive samples were antigen-positive, thus giving an overall sensitivity of 62.5%. The overall specificity was 96.0%, when all the culture-negative samples were combined. The results thus demonstrate that the antigen detection assay can provide a rapid supplemental information for the diagnosis of pulmonary diagnosis.
Antibodies ; Diagnosis* ; Diagnostic Tests, Routine ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mycobacterium tuberculosis* ; Mycobacterium* ; Sensitivity and Specificity ; Sputum* ; Tuberculosis ; Tuberculosis, Pulmonary*

BCG has been one of the vehicles for multi-recombinant vaccine. However, low transformation efficiency of BCG with plasmid DNA hampered studies involving expression of foreign antigens in BCG. In an effort to determine the optimal conditions, this study was initiated to investigate factors involved in the transformation of BCG with a Mycobacterium-Escherichia coli shuttle vector, pYUB18, by electroporation. Mycobacterium bovis BCG (strain 1173P2) was grown in Middlebrook (M) 7H9 broth containing albumin-dextrose-catalase and 0.05% tween 80, and transformed BCG was grown in M7H10 agar containing kanamycin for counting viable cells. Pretreatment of BCG with 10 mM CaCl2 improved the transformation efficiency, but overnight incubation of BCG with 1% glycine did not. The transformation efficiency in BCG also varied depending on voltage, resistance, and DNA concentration. The maximum transformation efficiency was obtained when the infinity resistance, 12.5 Kv/cm, and 100 ng of DNA were used, and reached 1.4 x 10(5) CFU/microgram of plasmid DNA, which is about 3-100 times greater than those from previous reports. The transformation conditions described in this study, therefore, will give us a better position for employing BCG as a vehicle for developing multi-recombinant vaccines.
Calcium Chloride/pharmacology ; Comparative Study ; DNA/metabolism ; Electrophysiology ; Electroporation* ; Escherichia coli/genetics* ; Genetic Vectors* ; Glycine/pharmacology ; Mycobacterium/genetics* ; Mycobacterium bovis/genetics* ; Osmolar Concentration ; Transformation, Bacterial/physiology* ; Transformation, Bacterial/drug effects

BACKGROUND: Voiding disturbance is one of the frequent symptoms of myelitis. Clinical manifestations of urinary disturbance in the patients with myelitis are results of the specific neurological lesion of the spinal cord, but can be varied. To determine the relationship between the neurological spinal cord level and bladder dysfunction, we nalyzed the urodynamic study in 30 patients with myelitis with urinary symptoms. Neurological examination and spinal MR imaging determined the clinical and radiological spinal cord levels. Urodynamic findings were classified as detrusor hyperreflexia (DH), detrusor-external sphincter dyssynergia (DSD), detrusor areflexia/hyporeflexia (DA) or normal. We analyzed the correlation between the neurological spinal level and urodynamic findings. RESULTS: Among the 30 patients, 14 patients were revealed DA, 8 were DH, 2 were DSD, and 6 were normal. Of 22 cervicothoracic lesions, 9 had DA, 7 had DH and 2 had DSD, but 4 had normal urodynamic findings. Four lumbar and sacral lesions had DA. CONCLUSIONS: The Urodynamic study finding in the lumbosacral lesion revealed DA, but, in those with cervicothoracic lesion are varied including DA, DH, DSD and normal. Urodynamic evaluation may provide more information for management of neurourological dysfunction.
Ataxia ; Humans ; Magnetic Resonance Imaging ; Myelitis* ; Neurologic Examination ; Reflex, Abnormal ; Spinal Cord ; Urinary Bladder ; Urodynamics*

BACKGROUND: Voiding disturbance is one of the frequent symptoms of myelitis. Clinical manifestations of urinary disturbance in the patients with myelitis are results of the specific neurological lesion of the spinal cord, but can be varied. To determine the relationship between the neurological spinal cord level and bladder dysfunction, we nalyzed the urodynamic study in 30 patients with myelitis with urinary symptoms. Neurological examination and spinal MR imaging determined the clinical and radiological spinal cord levels. Urodynamic findings were classified as detrusor hyperreflexia (DH), detrusor-external sphincter dyssynergia (DSD), detrusor areflexia/hyporeflexia (DA) or normal. We analyzed the correlation between the neurological spinal level and urodynamic findings. RESULTS: Among the 30 patients, 14 patients were revealed DA, 8 were DH, 2 were DSD, and 6 were normal. Of 22 cervicothoracic lesions, 9 had DA, 7 had DH and 2 had DSD, but 4 had normal urodynamic findings. Four lumbar and sacral lesions had DA. CONCLUSIONS: The Urodynamic study finding in the lumbosacral lesion revealed DA, but, in those with cervicothoracic lesion are varied including DA, DH, DSD and normal. Urodynamic evaluation may provide more information for management of neurourological dysfunction.
Ataxia ; Humans ; Magnetic Resonance Imaging ; Myelitis* ; Neurologic Examination ; Reflex, Abnormal ; Spinal Cord ; Urinary Bladder ; Urodynamics*

Coral is a member of the class Anthozoa, which cause injury due to stings from their tentacles containing nematocytes or from their sharp-edged calcified outer skeletons. Sting from their nematocytes may cause coral poisoning and cause a rare type of contact dermatitis. A 24-year-old woman, who had been contacted to coral during skin-scuba diving in the Philippines 7 days before, was presented with multiple brownish plaques and patches with small vesicles on the dorsum of the left hand. Herein, we report a case of contact dermatitis to coral by the nematocyst stings of corals.
Anthozoa ; Bites and Stings ; Dermatitis, Contact ; Diving ; Female ; Hand ; Humans ; Nematocyst ; Philippines ; Skeleton ; Young Adult

Coral is a member of the class Anthozoa, which cause injury due to stings from their tentacles containing nematocytes or from their sharp-edged calcified outer skeletons. Sting from their nematocytes may cause coral poisoning and cause a rare type of contact dermatitis. A 24-year-old woman, who had been contacted to coral during skin-scuba diving in the Philippines 7 days before, was presented with multiple brownish plaques and patches with small vesicles on the dorsum of the left hand. Herein, we report a case of contact dermatitis to coral by the nematocyst stings of corals.
Anthozoa ; Bites and Stings ; Dermatitis, Contact ; Diving ; Female ; Hand ; Humans ; Nematocyst ; Philippines ; Skeleton ; Young Adult