Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the B subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.
Base Sequence ; Classification ; Complement System Proteins ; Diagnosis ; DNA ; DNA-Directed RNA Polymerases ; Mycobacterium tuberculosis ; Polymerase Chain Reaction* ; Polymorphism, Restriction Fragment Length* ; Residence Characteristics ; Sequence Analysis*

Child ; Cranial Nerves ; pathology ; Eyelids ; Female ; Humans ; Male ; Ophthalmoplegia ; etiology ; Polyradiculopathy ; complications

Cyclosporine(CsA) has been known to cause an endothelial dysfunction following its use as an immunosuppressive agent. On the other hand, the vascular endothelium has been recognized as an endocrine organ in its own right, i.e., it releases vasoactive factors such as nitric oxide(NO) and hyperpolarizing factor(EDHF). NO is synthesized by at least three isoforms of NO synthases(NOS), among which ecNOS is constitutively expressed in the endothelium. The principle of EDHF has not been determined. The present study was aimed at further investigating the mechanisms underlying the CsA-induced vasculopathy. Rats were treated with CsA (25mg/kg/day, subcutaneous) for one week and their thoracic aortae were isolated. Their changes of iso-metric tension in responses to acetylcholine, diazoxide, and high concentrations of calcium were recorded. The expression of ecNOS mRNA and protein was determined by reverse transcription-polymerase reaction and Western blot analysis, respectively. The acetylcholine-induced relaxation of the aortic rings was significantly diminished follawing the CsA-treatment, which was prevented by L-arginine supplemented along with the CsA-treatment. The relaxation of the thoracic aorta in response to either diazoxide or high concentrations of extracellular calcium was not affected by CsA-treatrnent. The vascular tissue contents of NO metabolites were significantly decreased following the CsA-treatment, which was also prevented by L-arginine-supple-mentation. Neither ecNOS mRNA nor protein expression was significantly altered following the CsA-treatment. It is suggested that CsA induces an endothelial dysfunction, which cannot be attributed to an altered role of EDHF, but to an impairment in L-arginine/NO pathway at the steps beyond NOS protein expression.
Acetylcholine ; Animals ; Aorta, Thoracic ; Arginine ; Blotting, Western ; Calcium ; Diazoxide ; Endothelium ; Endothelium, Vascular ; Hand ; Protein Isoforms ; Rats* ; Relaxation ; RNA, Messenger

No abstract available in English.
Humans ; Infant ; Sarcoma, Ewing ; Scapula

OBJECTIVES: This study evaluates the change of the left atrial appendage function before and after electrical cardioversion to understand the mechanism involved in systemic thromboembolism of atrial fibrillation. BACKGROUND: Systemic thromboembolism associated with electrical cardioversion of atrial fibrillation is thought to originate from the left atrium or left atrial appendage, or both.However, the mechanism involved is poorly understood. METHOD: We studied left atrial appendage function funcction with transesophageal echocardiography in 15 patients with atrial fibrillation before and after successful electrical cardioversion. We measured left atrial appendage emptying and filling velocities and left atrial appendage areas. Also we analysed the characteristic Dopper flow pattern of LAA. RESULT: Left atrial appendage emptying velocities before cardioversion were greater in patients without(32.0+/-13.2cm/sec) than in those with(21.4+/-7.6cm/sec) spontaneous echo contrast(SEC). Furthermore emptying velocities after cardioversion were significantly reduced group with (21.4+/-7.6 vs 12.2+/-9.6, p<0.05) and the groupwithout(32.0+/-13.2 vs 18.1+/-10.2, p<0.05)SEC. CONCLUSION: After electrical cardioversion for atrial fibrillation left atrial appendage function is impaired. These observations suggest that stunned left atrial appendage after cardioversion may predispose to thrombus formation, which may play a role in the mechanism involved in the occurrence of thromboembolism after cardioversion.
Atrial Appendage* ; Atrial Fibrillation* ; Echocardiography, Transesophageal* ; Electric Countershock* ; Heart Atria ; Humans ; Thromboembolism ; Thrombosis

INTRODUCTIONFor many years, ophthalmologists have looked at the optic nerve head to evaluate the status of glaucoma. Clinical examination of the optic nerve head and retinal nerve fibre layer (RNFL) is however, subjective and sometimes variable. Recent developments in computer-based imaging technologies have provided a means of obtaining quantitative measurements of the optic nerve head topography and peripapillary retinal nerve fibre layer thickness.

METHODSMultiple searches using Medline were carried out. Additional searches were made using reference lists of published papers and book chapters.

RESULTSStudies involving three imaging technologies namely, confocal scanning laser ophthalmoscopy, scanning laser polarimetry and optical coherence tomography were reviewed. Overall, these technologies were reproducible and demonstrate good sensitivity and specificity in the range of 70 to 80%. Inclusion of age and ethnicity normative database will make these technologies more effective in screening and diagnosis. Quantitative measurements provide useful parameters for monitoring of patients.

CONCLUSIONThere is no consensus on the best technology for assessing structural damage in glaucomatous optic neuropathy. Therefore, as with any investigation, the clinician should exercise clinical correlation and judgment before instituting the appropriate treatment.

Female ; Humans ; Middle Aged ; Ophthalmoscopy ; methods ; Optic Nerve ; Retina ; Sensitivity and Specificity ; Tomography, Optical ; methods ; Tomography, Optical Coherence

The gene encoding the 66 kilodalton (kDa) protein of Borrelia hermsii HS1 was cloned and sequenced. Chromosomal DNA was prepared from purified B. hermsii and used in construction of genomic library. The library was screened for positive clones by 314 bp DIG-labeled probe synthesized on the basis of the part of the sequence of B. hermsii. Positive clone was subcloned into p2ErO vector and was designated as pBH11. pBH11 were subcloned into pBluscript vector and were designated as pBH11-1 (500 bp), pBH11-2 (800 bp), pBH11-3 (600 bp) and pBH11-4 (800 bp). The plasmids were sequenced and determined the nucleotide sequence of p66. The open reading frame of the p66 consisted of 1803 base pairs coding for 600 amino acid protein. The basic information on the p66 gene of B. hermsii HS1 obtained from this study will be useful for further analysis and experiment of pathogenesis of the borrelia.
Base Pairing ; Base Sequence ; Borrelia* ; Clinical Coding ; Clone Cells* ; Cloning, Organism* ; DNA ; Genomic Library ; Open Reading Frames ; Plasmids

Fifteen women with symptomatic uterine myomas, diagnosed by clinical examination and confirmed by pelvic ultrasonography, were treated with intranasal insufflation of Nafarelin acetate, 200 micrograms, twice a day, for a total treatment period of 6 months. Treatment was evaluated with respect to subjective symptoms, changes in myoma size and uterine volume, variations in blood estradiol, FSH, LH and CA 125, and side effects. The following results were obtained: 1. All patients showed a marked reduction(p<0.001) in uterine volume. Before treatment volume measured 312.3+/-24.2cm3, after 12 weeks volume was 132.4+/-40.6cm3, and after 24 weeks it was 123.6+/-48.3cm3. 2. Blood estradiol and LH levels were decreased significantly(p<0.05) after treatment. The FSH level was decreased, but not significantly. 3. Blood CA 125 levels were increased 6 cases(40.0%) before treatment. The levels were normalized in all 6 cases after treatment. 4. Symptoms of uterine myoma disappeared or decreased. 5. Minor side effects, such as hot flushes, headache, general myalgia and fatigue, and vaginal dryness were encountered frequently although none necessitating discontinuation of treatment, These data suggest that Nafarelin acetate is useful for the treatment of uterine myoma. However, appropriate indications should be selected in the treatment of uterine myoma because the possible regrowth of uterine myoma after treatment limits the use of GnRH agonist.
Estradiol ; Fatigue ; Female ; Gonadotropin-Releasing Hormone* ; Headache ; Humans ; Insufflation ; Leiomyoma* ; Myalgia ; Myoma ; Nafarelin ; Ultrasonography

OBJECTIVE: The differential expression patterns of the two progesterone receptor (PR) isoforms, PRA and PRB were examined using immunohistochemistry and real time quantitative RT-PCR in normal and neoplastic ovarian tissues, and in cell lines derived from epithelial ovarian cancer and breast cancer as a positive control in order to further elucidate the possible involvement of progesterone in the development of ovarian neoplasia. METHODS: Breast cancer cell line MCF-7 and ovarian cancer cell line SNU-8 were seeded to 24-well plate with 5 X 10(5) cell/well and incubated overnight. Those cell lines were treated with 17-beta-estradiol and incubated for another 24 hrs. RNA were purified for RT-PCR and whole cell prepared total proteins were subject to immunoblot with DAB-staining. DNA ladder pattern and flowcytometry were studied to evaluate progestins induced apoptosis in the ovarian epithelium. RESULTS: The median H-scores for PR isoforms in normal (n=8), benign (n=10), borderline (n=8) and malignant (n=24) ovarian tissues were as follows; PRA: 194.0, 171.0, 49.5, 0.0 (P<0.05), and PRB: 175.0, 180.5, 251.5, 168.5, respectively. In ovarian cancer cell line (SNU-8), PRB/PRAB mRNA ratio was not increased by 17-beta-estradiol, but that ratio was increased by 17-beta-estradiol in breast cancer cell line (MCF 7). Immunoblotting analysis demonstrated that PRB protein expression was markedly up- regulated in SNU-8, whereas the PRA and PRB isoforms both appeared to be increased in MCF-7. DNA ladder pattern was increased in dose and time related exposure of progestins and apoptotic cells were markedly increased during exposure progestins in ovarian cancer cell line were found by flowcytometry. CONCLUSION: These results suggest that down-regulation of PRA is associated with the development of ovarian epithelial carcinoma. Progestins can activate the apoptotic pathway in the ovarian epithelium for protection of normal tissues from neoplatic transformation suggests that progestins deserve further evaluation as potential ovarian cancer preventive agents.
Apoptosis ; Breast Neoplasms ; Cell Line ; DNA ; Down-Regulation ; Epithelium ; Female ; Immunoblotting ; Immunohistochemistry ; Ovarian Neoplasms ; Ovary* ; Progesterone* ; Progestins ; Protein Isoforms* ; Receptors, Progesterone* ; RNA ; RNA, Messenger

No abstract available.