BACKGROUND: Although there are many methods including AFB smear and culture, and the analysis of pleural fluid in the etiological diagnosis of pleural effusion, it is sometimes difficult to confirm a diagnosis especially in cases of incomplete pleural biopsies. Moreover, the high incidence of tuberculous pleuritis in young people caused confusion in the differential diagnosis of pleural effusion in Korea. The pathognomonic finding of tuberculous pleuritis in pleural biopsy is chronic granulomatous pleuritis (CGP) with caseous necrosis. But a biopsy does not always provide a definitive diagnosis, which shows in only 60~70% of all biopsies, because of either limitations in blind biopsies or inadequate specimens. An adequate biopsy also gives only limited information, such as chronic or nonspecific pleuritis. METHODS: We compared the clinical diagnosis, pathologic findings and detection of mycobacterial DNA using nested PCR of pleural biopsy tissues. We carried out the nested PCR for IS6110 insertion sequence of Mycobacterium tuberculosis using outer primer IS-1/IS-2 (5'-AGGCGTTGGTTCGCGAGGG-3' /5'-TGATGACGCCCTCGTTGCC-3') and inner primer IS-3/IS-4 (5'-CCAACCCGCTCGGTCTCAA-3' /5'-ACCGATGGACTGGTCACCC-3') in 52 pleural biopsy tissues which were pathologically diagnosed as tuberculous pleuritis, malignant pleuritis or non-specific pleuritis. RESULTS: Five (71.4%) of 7 cases clinically and pathologically confirmed tuberculous pleuritis diagnosed as chronic granulomatous pleuritis (CGP) with caseous necrosis revealed positive in nested PCR for M. tuberculosis. Seven (36.8%) of 19 cases diagnosed as CGP without caseous necrosis were positive. However, only 3 (25%) of 12 cases diagnosed as non-specific chronic pleuritis were positive by PCR for M. tuberculosis. Neither congestive heart failure nor malignancies with pleurisy showed a positive reaction. CONCLUSION: In this study, pathologic findings were significantly associated with the detection rate of mycobacterial DNA. And, even in patients with nonspecific or chronic inflammatory pleuritis, mycobacterial DNA could be detected by using nested PCR in pleural biopsy tissue with good specificity. Detection of mycobacterial DNA in pleural tissue might provide additional information for etiological diagnosis in patients with pleural effusion.
Biopsy ; Comparative Study ; DNA, Bacterial/*isolation & purification ; Human ; Lung/microbiology/*pathology ; Mycobacterium tuberculosis/*genetics/isolation & purification ; *Polymerase Chain Reaction ; Support, Non-U.S. Gov't ; Tuberculosis, Pulmonary/*diagnosis/genetics/microbiology

Epstein-Barr Virus Infections ; Genes, p53 ; genetics ; Humans ; Lung Diseases, Interstitial ; pathology ; Lung Neoplasms ; genetics ; pathology ; virology ; Mutation ; Pleural Neoplasms ; genetics ; pathology ; Pulmonary Disease, Chronic Obstructive ; pathology ; Severe Acute Respiratory Syndrome ; pathology ; Tuberculosis, Pulmonary ; pathology

PURPOSE: Guinea pig is one of the most suitable animal models for Mycobacterium tuberculosis (M. tb) infection since it shows similarities to pulmonary infection in humans. Although guinea pig shows hematogenous spread of M. tb infection into the whole body, immunological studies have mainly focused on granulomatous tissues in lungs and spleens. In order to investigate the time-course of disease pathogenesis and immunological profiles in each infected organ, we performed the following approaches with guinea pigs experimentally infected with M. tb over a 22-week post-infection period. MATERIALS AND METHODS: We examined body weight changes, M. tb growth curve, cytokine gene expression (IFN-gamma and TNF-alpha), and histopathology in liver, spleen, lungs and lymph nodes of infected guinea pigs. RESULTS: The body weights of infected guinea pigs did not increase as much as uninfected ones and the number of M. tb bacilli in their organs increased except bronchotracheal lymph node during the experimental period. The gene expression of IFN-gamma and TNF-alpha was induced between 3 and 6 weeks of infection; however, kinetic profiles of cytokine gene expression showed heterogeneity among organs over the study period. Histophathologically granulomatous lesions were developed in all four organs of infected guinea pigs. CONCLUSION: Although IFN-gamma and TNF-alpha gene expression profiles showed heterogeneity, the granuloma formation was clearly observed in every organ regardless of whether the number of bacilli increased or decreased. However, this protective immunity was accompanied with severe tissue damage in all four organs, which may lead to the death of guinea pigs.
Animals ; Body Weight ; *Disease Progression ; Female ; Gene Expression ; Gene Expression Regulation ; Guinea Pigs ; Interferon-gamma/genetics/*metabolism ; Kinetics ; Liver/metabolism/pathology ; Lung/metabolism/pathology ; Lymph Nodes/metabolism/pathology ; Mycobacterium tuberculosis ; Spleen/metabolism/pathology ; Tuberculosis/*genetics/pathology ; Tumor Necrosis Factor-alpha/genetics/*metabolism

A case of tuberculosis is reported in an eight-year-old, male, elk (Cervus elaphus nelsoni). The elk showed severe coughing, respiratory distress, abdominal breathing, anorexia, and severe progressive emaciation in the elk farm. At necropsy, the elk appeared in poor body condition. Mild enlargement of retropharyngeal and submandibular lymph node was observed in the head. Diffuse fibrinous pleuritis and purple red lobar pneumonia were found in the thorax. Well demarcated numerous dark yellow discrete or confluent nodules from 0.3 to 2 cm in diameter were scattered in the whole lung. Bronchial and mediastinal lymph nodes were also enlarged. Histopathologically, lungs had typical classical tuberculous granulomas, multiple abscesses, and numerous macrophages and Langhans giant cells infiltration in alveolar lumen. In the lymph nodes, there were small clusters of necrosis and infiltration of numerous macrophages, epithelioid cells, and Langhans giant cells. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. According to this study, there are differences of the histopathologic lesions and the numbers of acid-fast bacilli in the lesions between this elk and cattle. Mycobacterium bovis was confirmed as a causative agent in this elk using bacterial isolation, biochemical characteristics, and PCR technique. The isolate was negative for niacin test, nitrate reductase, and pyrazinamidase. This is a first report for bovine tuberculosis of farmed elk in Asia.
Animals ; DNA, Bacterial/chemistry/genetics ; Deer/*microbiology ; Fatal Outcome ; Korea ; Lung/microbiology/pathology ; Male ; Mycobacterium bovis/genetics/*isolation&purification ; Polymerase Chain Reaction ; Tuberculosis/microbiology/*veterinary

OBJECTIVETo study the effection of suppression murine melanoma growth by Intratumor injection of recombinant attenuated salmonella carrying heat shock protein 70 and herpes simplex virus thymidine kinase genes.

METHODSPlasmids PCMV-mtHSP70-IRES-TK were electro-transferred into salmonella typhimurium SL7207 to construct recombinant salmonella typhimurium. In vivo, Recombinant bacteria were injected into the mouse melanoma and the antitumor effection was observed. The survival period was recorded and safety analysis for this vaccine in each group.

RESULTSIn vivo, the mtHSP70/HSV-tk recombinant bacteria can suppress tumor growth significantly and extend survival. After recombinant Salmonella, 10(9) CFU/mL, was administered as an intratumoral injection, No diarrhea were observed. During therapy, body weight did not change markedly.

CONCLUSIONResults of the animal experiment suggests intratumor injection of recombinant attenuated salmonella typhimurium containing mtHSP70 and HSV-tk genes, has targeting ability against B16 tumor cell and could significantly inhibit tumor growth .

Animals ; Bacterial Proteins ; genetics ; immunology ; Cancer Vaccines ; genetics ; immunology ; pharmacology ; Genetic Therapy ; methods ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Melanoma, Experimental ; microbiology ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Mycobacterium tuberculosis ; genetics ; Salmonella typhimurium ; genetics ; immunology ; Simplexvirus ; enzymology ; genetics ; Skin Neoplasms ; therapy ; Thymidine Kinase ; genetics ; immunology ; Vaccines, Attenuated ; genetics ; immunology ; pharmacology ; Vaccines, DNA ; genetics ; immunology ; pharmacology

Tuberculous liver abscesses are rare. Paradoxical response in tuberculosis is common and occurred between 2 weeks and 12 weeks after anti-tuberculous medication. We report here a case of tuberculous liver abscess that developed in a paradoxical response during chemotherapy for tuberculous peritonitis in a 23-year-old male. He was hospitalized, complaining of ascites, epigastric pain. He was diagnosed tuberculous peritonitis by expiratory laparoscopic biopsy and took medication for tuberculosis. After 2 months, a hepatic lesion was detected with CT scan incidentally. Chronic granulomatous inflammation was seen in ultrasound-guided liver biopsy, and tuberculous liver abscess was diasnosed. It was considered as paradoxical response, rather than treatment failure or other else because clinical symptoms of peritoneal tuberculosis and CT scan improved. After continuing initial anti-tuberculous medication, he was successfully treated. Herein, we report a case of tuberculous liver abscess as paradoxical response while treating peritoneal tuberculosis without changing anti-tuberculous treatment regimen.
Antitubercular Agents/*adverse effects/*therapeutic use ; DNA, Bacterial/analysis ; Humans ; Laparoscopy ; Liver/pathology/ultrasonography ; Liver Abscess/*chemically induced/*diagnosis/microbiology ; Male ; Mycobacterium tuberculosis/genetics/isolation & purification ; Necrosis/pathology ; Peritoneum/pathology ; Peritonitis, Tuberculous/*drug therapy ; Tomography, X-Ray Computed ; Tuberculosis/*diagnosis/microbiology ; Young Adult

Adult ; Aged ; Aged, 80 and over ; Buruli Ulcer ; pathology ; surgery ; DNA, Bacterial ; analysis ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mycobacterium ulcerans ; genetics ; isolation & purification ; Tuberculosis, Cutaneous ; pathology

OBJECTIVETo study the role of Mycobacterium tuberculosis in the pathogenesis of sarcoidosis.

METHODSArchival material of 22 patients with a histologic diagnosis of sarcoidosis were retrieved. Real-time fluorescent polymerase chain reaction (PCR) was used to detect DNA fragments of the complex-specific insertion sequence IS6110 of Mycobacterium tuberculosis in formalin-fixed and paraffin-embedded biopsy samples.

RESULTSAmong the 22 samples studied, Mycobacterium tuberculosis DNA was detected in 11 cases. The sequence of PCR amplified IS6110 DNA fragments completely matched with the related sequence in Mycobacterium tuberculosis gene.

CONCLUSIONSMycobacterium tuberculosis DNA is identified in a certain proportion of patients with a clinicopathologic diagnosis of sarcoidosis. Mycobacterium tuberculosis may be an important etiologic agent, at least in some of these patients.

Adult ; DNA, Bacterial ; analysis ; Female ; Fluorescence ; Follow-Up Studies ; Humans ; Lymph Nodes ; microbiology ; pathology ; Male ; Middle Aged ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Sarcoidosis ; microbiology ; pathology

OBJECTIVETo investigate the relationship between the polymorphisms of UGT1A6 genes and anti-tuberculosis drug induced hepatic-injury (ADIH).

METHODS202 cases and 239 controls were collected and a case-control study was conducted. Information on related risk factors of tuberculosis was collected. The genotypes of UGT1A6-19T/G, UGT1A6-308C/A and UGT1A6-541A/G genetic polymorphisms were detected by polymerase chain reaction and restriction fragment length polymorphism technique (PCR-RFLP) in patients received anti-tuberculosis therapy. The Hha I, Dpn II and Nsi I enzyme were employed. Univariate and multivariate conditional logistic analyses were conducted using SPSS13.0 for windows software.

RESULTS

RESULTSThe allele frequency of gene UGT1A6-19T/T, UGT1A6-19T/G, UGT1A6-19G/G, GT1A6-308C/C, UGT1A6-308C/A, UGT1A6-308A/A, UGT1A6-541AA, UGT1A6-541A/G and UGT1A6-541G/G in ADIH group were 51.5%, 39.6%, 8.9%, 52.0%, 40.6%, 7.4%, 57.9%, 33.7%, 8.4% and 71.1%, 25.5%, 3.3%, 79.1%, 19.2%, 1.7%, 79.5%, 19.2%, 1.3% in control group, respectively. Univariate analysis demonstrated that the frequency of UGT1A6-19T/G, UGT1A6-308C/A and UGT1A6-541A/G genotype in cases were significantly higher than that in controls (P less than 0.05).

CONCLUSIONA positive association is found between UGT1A6 genotype and the occurrence of ADIH. The synergetic effect is proved on susceptibility to pulmonary tuberculosis between UGT1A6 mutant genotypes.

Adult ; Aged ; Antitubercular Agents ; adverse effects ; Case-Control Studies ; Chemical and Drug Induced Liver Injury ; genetics ; Female ; Genotype ; Glucuronosyltransferase ; genetics ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Polymorphism, Genetic ; Tuberculosis, Pulmonary ; drug therapy

BACKGROUND: Conventional acid-fast bacilli (AFB) staining cannot differentiate viable from dead cells. Propidium monoazide (PMA) is a photoreactive DNA-binding dye that inhibits PCR amplification by DNA modification. We evaluated whether PMA real-time PCR is suitable for the early detection of viable Mycobacterium tuberculosis (MTB) in clinical respiratory specimens. METHODS: A total of 15 diluted suspensions from 5 clinical MTB isolates were quadruplicated and subjected to PMA treatment and/or heat inactivation. Eighty-three AFB-positive sputum samples were also tested to compare the DeltaC(T) values (C(T) value in PMA-treated sputum samples-C(T) value in non-PMA-treated sputum samples) between culture-positive and culture-negative specimens. Real-time PCR was performed using Anyplex MTB/NTM Real-Time Detection (Seegene, Korea), and the C(T) value changes after PMA treatment were compared between culture-positive and culture-negative groups. RESULTS: In MTB suspensions, the increase in the C(T) value after PMA treatment was significant in dead cells (P=0.0001) but not in live cells (P=0.1070). In 14 culture-negative sputum samples, the median DeltaC(T) value was 5.3 (95% confidence interval [CI], 4.1-8.2; P<0.0001), whereas that in 69 culture-positive sputum samples was 1.1 (95% CI, 0.7-2.0). In the ROC curve analysis, the cutoff DeltaC(T) value for maximum sensitivity (89.9%) and specificity (85.7%) for differentiating dead from live cells was 3.4. CONCLUSIONS: PMA real-time PCR is a useful approach for differentiating dead from live bacilli in AFB smear-positive sputum samples.
Adult ; Aged ; Area Under Curve ; Azides/*chemistry ; DNA, Bacterial/*analysis ; Female ; Humans ; Lung Diseases/diagnosis/*microbiology/pathology ; Male ; Middle Aged ; Mycobacterium tuberculosis/genetics/*isolation & purification ; Pilot Projects ; Propidium/*analogs & derivatives/chemistry ; ROC Curve ; *Real-Time Polymerase Chain Reaction ; Sputum/microbiology ; Tuberculosis/*diagnosis/microbiology