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

No abstract available.
Antibodies* ; Coxiella burnetii* ; Coxiella* ; Korea* ; Prevalence*

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*

Taxol is an active chemotherapeutic agent against a variety of solid tumors and a potentially useful drug for augmenting the cytotoxic action of radiotherapy against certain cancers. Taxol blocks cells in the mitotic phase of cell cycle. The aim of this study was to define the in vivo response of rapidly dividing cells of the small intestinal mucosa to taxol. We studied the numbers of apoptotic and mitotic cells and the expression of bcl-2 and p53 in rat jejunal crypt cells at 1, 2, 4, 8, 12, 16, and 24 hours and 3 and 5 days after intraperitoneal injection of taxol. Mitosis peaked at 2 and 4 hours and 12 and 16 hours. Apoptosis peaked at 16 hours and returned to normal after five days. The glands in crypts showed marked distortion with atypical lining cells after three days, which returned to normal at 5 days. bcl-2 expression was markedly decreased at 8 to 24 hours and subnormally recovered after three to five days. p53 showed no significant changes throughout. The histopathological changes in small intestine due to taxol were transient with complete recovery. bcl-2 expression was inversely corresponded to numbers of apoptosis. The changes were p53 independent. Further studies to understand the conditions that maximize the cell-cycle modulating effects of taxol cl-may greatly enhance its anti-tumor effectiveness.
Animals ; Apoptosis ; Cell Cycle ; Injections, Intraperitoneal ; Intestinal Mucosa ; Intestine, Small* ; Mitosis ; Paclitaxel ; Radiotherapy ; Rats*

Dieulafoy ulcer is an unusual cause of massive, recurrent and frequently fatal gastrointestinal hemorrhage that results from erosion of abnormally large submucosal artery. Although the lesion has been found throughout the gastrointestinal tract, it most commonly occurs in the proximal stomach. Diagnosis depends on the observation of protruding and eroded artery with pulsatile bleeding or adherent thrombus by endoscopy. Even during active bleeding, the endoseopic examination can be negative if intraluminal blood or clots obscure the source of bleeding. If the bleeding has stopped, the small mucosal lesion can be easily overlooked. Unlike peptic ulceration, there is no excavation of the mucosa. A 76-year-old man presented with massive hematemesis and melena. The patient had no previous history of peptic ulcer disease. He did not drink alcohol and use aspirin or NSAIDs. Physical examination revealed a pale, severely diaphoretic male with hypotension and melenic stools. He was found to have hemoglobin 4.0 g/dL and hematocrit 12.7%. We performed emergency endoscopy which showed a pulsatile and bleeding exposed artery without evidence of surrounding ulcerative lesion on the posterior wall of upper body of stomach. Endoscopic ligation using O ring of Stiegman-Goff endoscopic ligator kit was done successfully and the bleeding stopped immediately after ligation. Ten days after treatment, endoscopy showed artificial ulcerative lesion on previous ligated site and no evidence of bleeding. Another endoscopy four days later revealed healing ulcerative lesion. After improvement, the patient was discharged and rebleeding has not occurred to date.
Aged ; Anti-Inflammatory Agents, Non-Steroidal ; Arteries ; Aspirin ; Cytochrome P-450 CYP1A1 ; Diagnosis ; Emergencies ; Endoscopy ; Gastrointestinal Hemorrhage ; Gastrointestinal Tract ; Hematemesis ; Hematocrit ; Hemorrhage ; Humans ; Hypotension ; Ligation* ; Male ; Melena ; Mucous Membrane ; Peptic Ulcer ; Physical Examination ; Stomach ; Thrombosis ; Ulcer*

OBJECTIVE: The implantation failure after embryo-transfer (ET) is a major continuing problem in in vitro fertilization (IVF). This study was undertaken to determine the effectiveness of intravenous immunoglobulin for treatment of individuals experiencing repeated unexplained in vitro fertilization-embryo transfer (IVF-ET) failure. METHODS: A total of nine consecutive infertile patients who failed to become pregnant after previous IVF-ET replacing at least three or more normal developed embryos each were included in our study. During the subsequent new IVF-ET cycle, each women received intravenous immunoglobulin 500mg/kg before the embryo transfer. RESULTS: Only one implantation occurred. There were no remarkable side effects. A specific effect of intravenous immunoglobulin for patients with repeated IVF-ET failure could not be demonstrated. CONCLUSION: High-dose intravenous immunoglobulin may not be useful for patients with repeated failure of embryo transfer.
Embryo Transfer ; Embryonic Structures ; Female ; Fertilization in Vitro ; Humans ; Immunoglobulins*

The diagnosis and assessment of pulmonary arterial hypertension (PAH) is a rapidly evolving area, with changes occurring in the definition of the disease, screening and diagnostic techniques, and staging and follow-up assessment. After 4th World Symposium on pulmonary hypertension (PH) which took place in Dana Point in early 2008, the definition of PH has been simplified (mean pulmonary artery pressure > or =25 mmHg) based on currently available evidences. The diagnosis of PH involves two stages: detection (determining the cause of a patient's symptoms, or to detect the presence of PAH in a high-risk patient) and characterization (determining the specific clinical context of the PH, including causal factors, associated diseases or substrates and hemodynamic perturbations). Echocardiography and right heart catheterization have been main diagnostic method in PAH. Also, there has been progress in imaging techniques and biomarkers used to screen patients for the disease and to follow up their response to therapy. Useful tools to predict outcome include functional class, exercise capacity, pulmonary hemodynamics, acute vasoreactivity, right ventricular function, as well as brain natriuretic peptide, endothelin-1, and so on. As new therapies have been developed for PAH, screening, prompt diagnosis, and accurate assessment of disease severity have become increasingly important. A clear definition of PH and the development of a rational approach to diagnostic assessment and follow-up using both conventional and new tools will be essential to advance proper treatment of patients.
Biomarkers ; Cardiac Catheterization ; Cardiac Catheters ; Echocardiography ; Endothelin-1 ; Follow-Up Studies ; Hemodynamics ; Humans ; Hydrogen-Ion Concentration ; Hypertension ; Hypertension, Pulmonary ; Hypogonadism ; Mass Screening ; Mitochondrial Diseases ; Natriuretic Peptide, Brain ; Ophthalmoplegia ; Pulmonary Artery ; Ventricular Function, Right

Blue toe syndrome is characterized by tissue ischemia secondary to cholesterol crystal or atherothrombotic embolization. It leads to the occlusion of small vessels. The treatment option is usually surgery for most causes of blue toe syndrome. However, endovascular aortic repair by aorto-iliac stent graft become more and more popular because of its effectiveness and its less invasive characteristic. We present a 57-year-old man who suffered from blue toes syndrome on both legs caused by embolizing aorto-iliac lesions. Successful Endurant stent graft (Medtronic Vascular, Santa Rosa, CA, USA) was performed on infrarenal abdominal aorta and on proximal portion of right and left common iliac artery.
Aorta, Abdominal ; Blood Vessel Prosthesis ; Blue Toe Syndrome* ; Cholesterol ; Humans ; Iliac Artery ; Ischemia ; Leg ; Middle Aged ; Rosa ; Thromboembolism ; Toes

Familial juvenile polyposis is a rare intestinal polyposis characterized by the occurrence of multiple juvenile polyps in the gastrointestinal tract. We report a case of familial juvenile polyposis in a 17-year-old man with a review of the literature. This patient underwent total colectomy due to a 6 years history of rectal bleeding. Grossly, the colon showed 36 variable sized pedunculated polyps, measuring 2.5cm x 2cm from the largest size and 0.2cm x 0.2cm to the smallest size. Histologically, the polyps consisted of cystically dilated glands, lined by normal colonic epithelial cells, scattered in loose, edematous stroma containing inflammatory cell infiltration. There were no areas of tubular adenoma or malignancy in any of the polyp.
Adenoma ; Adolescent ; Colectomy ; Colon ; Epithelial Cells ; Gastrointestinal Tract ; Hemorrhage ; Humans ; Intestinal Polyposis ; Polyps

BACKGROUND: Histologic progressive changes of bile duct epithelium with hyperplasia, dysplasia and cholangiocarcinoma could be caused by hepatolithiasis. To be clarified as a neoplastic process, this histologic process should be evaluated with various aspects of cell biology. METHODS: Immunohistochemical study of proliferating cell nuclear antigen (PCNA) was performed on 45 cases (10; normal, 15; hyperplasia, 14; low-grade dysplasia:LGD, 6; high-grade dysplasia: HGD) of hepatolithiasis and 10 cases (all HGD) of hepatolithiasis with cholangiocarcinoma. RESULTS: In the hepatolithiasis, mean PCNA labelling indices (LI) of normal, hyperplasia, LGD and HGD of major intrahepatic bile duct epithelium were 24.5+/-4.3, 51.5+/-0.1, 62.0+/-.4 and 84.7+/-.3, respectively and gradually increased. Mean LI of PCNA in HGD of major intrahepatic bile duct epithelium of hepatolithiasis with cholangiocarcinoma was 68.7+/-.7, which was similar to that of LGD in hepatolithiasis without cholangiocarcinoma. CONCLUSIONS: Histologic transformation through hyperplasia, dysplasia and carcinoma in major intrahepatic bile duct epithelium of hepatolithiasis may be a neoplastic process if these histologic changes are evaluated in the cellular proliferation aspect.
Bile Ducts ; Bile Ducts, Intrahepatic* ; Cell Proliferation ; Cholangiocarcinoma* ; Epithelium* ; Hyperplasia ; Liver* ; Proliferating Cell Nuclear Antigen*