TUBERCULOSIS

TUBERCULOSIS

Diagnosis, Diagnostics, Immunodiagnosis & Immunodiagnostics:

ABSTRACT

January 2003

6311. Anuradha S, Ravinder Kaur, Singh NP, Baveja UK. Serodiagnosis of extra pulmonary tuberculosis using A-60 antigen. J Common Dis. 2001; 33(1): 12-16.

Abstract: Diagnosis of extrapulmonary tuberculosis including neurotuberculosis is difficult because of the low yield of culture positivity for Mycobacterium tuberculosis (M. tb.). Serodiagnosis has emerged as a useful aid to the diagnosis of extrapulmonary tuberculosis. The utility and efficacy of detection of antimycobacterial antibodies to A-60 antigen in serum and/or cerebrospinal fluid (CSF) was analysed in 100 patients-neurotuberculosis-72, abdominal tuberculosis-12 and others-16. The overall positivity rate for the test was 75%. The positivity rate of the test in serum and/or CSF was 79.2% (57 of 72) in neurotuberculosis and 62.5% (10 of 16) for other forms of extrapulmonary tuberculosis. The positivity rate for antimycobacterial antibodies was higher for patients with tubercular meningitis –94.7%. To conclude, testing for antimycobacterial antibodies to A-60 antigen is a useful adjunct in the diagnosis of extrapulmonary tuberculosis especially neurotuberculosis.

6312. Baker CA, Cartwright CP, Williams DN, Nelson SM, Peterson PK. Early detection of central nervous system tuberculosis with the gen-probe nucleic Acid amplification assay: utility in an inner city hospital. Clin Infect Dis. 2002 Aug 1;35(3):339-42.

Central nervous system tuberculosis is a serious clinical problem, the treatment of which is sometimes hampered by delayed diagnosis. We investigated the utility of the Gen-Probe nucleic acid amplification assay for the rapid diagnosis of tuberculous meningitis and as a noninvasive method of identifying intracranial tuberculoma.

6313. Burgess LJ, Reuter H, Carstens ME, Taljaard JJ, Doubell AF. The use of adenosine deaminase and interferon-gamma as diagnostic tools for tuberculous pericarditis. Chest. 2002 Sep;122(3):900-5.

BACKGROUND: Traditional diagnostic tests for pericardial tuberculosis (TB) are insensitive and often require long culture periods, and this has led to more emphasis being placed on biochemical tests such as the pericardial adenosine deaminase (ADA) test. However, controversy exists as to its diagnostic utility. In addition, the use of interferon (IFN)-gamma, which is a reliable indicator of pleural and peritoneal TB, has not been explored in pericardial effusions. We investigated ADA and IFN-gamma levels in pericardial effusions of different etiologies. METHODS AND RESULTS: A prospective study was carried out from February 1995 to February 1998 at Tygerberg Hospital (South Africa), with pericardial taps being performed under echocardiographic guidance. During this period, 110 consecutive patients presenting with large pericardial effusions were included in the study. Diagnoses were made according to predetermined criteria, and they included TB (n = 64), malignancy (n = 12), nontuberculous infections (n = 5), other effusions (n = 19), and effusions of uncertain origin (n = 10). The median ADA level in the tuberculous group was 71.7 U/L (range, 10.3 to 303.6 U/L), which was significantly higher than that in any other group (p < 0.05). With a cutoff level for ADA activity of 30 U/L, sensitivity was 94%, specificity was 68%, and positive predictive value was 80%. IFN-gamma levels were determined in 30 subjects. The median IFN-gamma concentration in the tuberculous group was > 1,000 pg/L, which was significantly higher than in any other diagnostic group (p < 0.0005). A cutoff value of 200 pg/L for IFN-gamma resulted in a sensitivity and specificity of 100% for the diagnosis of pericardial TB. CONCLUSION: Pericardial fluid levels of ADA and IFN-gamma are useful in the diagnosis of tuberculous pericarditis.

6314. Cavusoglu C, Cicek-Saydam C, Karasu Z, Karaca Y, Ozkahya M, Toz H, Tokat Y, Bilgic A. Mycobacterium tuberculosis infection and laboratory diagnosis in solid-organ transplant recipients. Clin Transplant. 2002 Aug;16(4):257-61.

Tuberculosis (TB) is an unusual infection in transplant recipients. We evaluated (i) the frequency of TB, (ii) the duration to develop the TB infection, and (iii) clinical consequences, in 380 solid-organ recipients from January 1995 to December 2000. A total of 10 (2.63%) patients (eight renal, two liver transplant recipients) were found to have post-transplantation TB. The frequency of TB in this patient population is 8.5-fold higher than the prevalance in the general Turkish population. Tuberculosis developed within 2-33 months after transplantation, with a median of 15 months. In all of these 10 patients, Mycobacterium tuberculosis (MTB) was isolated from the culture. All the patients continued to have low dose immunosuppressive treatment, and also quadriple antituberculosis treatment [isoniazid (INH), rifampin (RIF), pyrazinamide (PRZ) and ethambutol (ETB)] has been given. The two recipients had died of disseminated form of TB. Relapse was detected in one patient 6 months after the completion of the treatment. As post-transplant TB infection develops mostly within the first year after transplantation, clinicians should be more careful for early and fast diagnosis and treatment should be started immediately.

6315. Chandramuki A, Lyashchenko K, Kumari HB, Khanna N, Brusasca P, Gourie-Devi M, Satishchandra P, Shankar SK, Ravi V, Alcabes P, Kanaujia GV, Gennaro ML. Detection of antibody to Mycobacterium tuberculosis protein antigens in the cerebrospinal fluid of patients with tuberculous meningitis. J Infect Dis. 2002 Sep 1;186(5):678-83.

Antibodies against Mycobacterium tuberculosis antigens were detected by enzyme-linked immunosorbent assay in cerebrospinal fluid (CSF) samples obtained from 442 patients with tuberculous meningitis (TBM) and 102 control patients. Antibodies were found in the CSF of 87% of patients with clinical (culture-negative) TBM, 72% of patients with culture-positive TBM, and 65% of patients with autopsy-proven TBM. That anti-M. tuberculosis antibodies were detected in the CSF of patients with clinically diagnosed cases more frequently than in patients with culture-positive cases suggests that the detection of antibodies in CSF tends to decrease as bacillary load increases. Of the patients with clinical TBM who were coinfected with human immunodeficiency virus (HIV), 70% exhibited anti-M. tuberculosis antibody in CSF, which suggests that antibody responses in this group were substantially weaker than those in HIV-negative patients with clinical TBM. Some groups showed a stronger response to certain antigens, which suggests that antigen recognition patterns may be specific for the stage of disease.

6316. Conde MB, Marinho SR, Pereira Mde F, Lapa e Silva JR, Saad MH, Sales CL, Ho JL, Kritski AL. The usefulness of serum adenosine deaminase 2 (ADA2) activity in adults for the diagnosis of pulmonary tuberculosis. Respir Med. 2002 Aug;96(8):607-10.

Rapid diagnosis of Mycobacterium tuberculosis remains an obstacle for therapy of tuberculosis (TB). Adenosine deaminase isoform 2 (ADA2) is produced by activated macrophages and has been used for diagnosis of TB from extra-pulmonary sites. However, few studies adequately address whether serum ADA2 activity is useful for diagnosis of active pulmonary tuberculosis (PTB). We prospectively measured serum ADA2 activity in 110 patients with pulmonary disease (65 cases with active PTB and 45 cases with other respiratory diseases) and 78 healthy volunteers (eight with tuberculin skin test positive). The serum ADA2 for the diagnosis of PTB had the sensitivity of 36.9%, the specificity of 84.5%, the positive predictive value of 10.9% and the negative predictive value of 96.2%. We concluded that serum ADA2 activity is neither useful to diagnosis of active PTB nor to differentiate from other respiratory diseases.

6317. Dhar S; Dhar S. Histopathological features of granulomatous skin diseases : an analysis of 22 skin biopsies Indian Journal of Dermatology . 2002; 47(2): 88-90.

ABSTRACT: In the present study, skin biopsies were analysed for histopathological (HP) changes in 22 patients with various granulomatous dermatoses. In 6 specimens, HP features were diagnostic of BT leprosy, in 1 each of BB, BL and histoid LL. The diagnosis was lupus vulgaris (LV) in 6 biopsies, tuberculosis verrucosa cutis (TBVC) in 2, sarcoidosis in 3 and sporotrichosis in remaining 2. The study reiterated the usefulness of HP examination of all suspected cases of granulomatous skin diseases.

6318. Dilmac A, Ucoluk GO, Ugurman F, Gozu A, Akkalyoncu B, Eryilmaz T, Samurkasoglu B. The diagnostic value of adenosine deaminase activity in sputum in pulmonary tuberculosis. Respir Med. 2002 Aug;96(8):632-4.

This study was carried out in Ataturk Chest Diseases and Surgery Center. It's aim to determine and compare sputum adenosine deaminase (ADA) activity in pulmonary tuberculosis (tb), lung cancer and chronic obstructive pulmonary disease (COPD) patients in order to assess its diagnostic value. PATIENTS AND METHOD: Eighty-four patients (25 tb, 30 lung cancer and 29 COPD) were included in the study. ADA activity in sputum and serum was measured. Sputum ADA activities of tb patients were significantly higher than the other two groups (P < 0.05). Sputum/serum ADA ratios were similar in all groups. Sputum ADA activities between 150 and 200 U/L were the measurements with the best test performance according to the ROC curve. Sensitivity, specificity, positive predictive value, and negative predictive value were 44.0, 86.4, 57.8, 78.4% for 150 U/L and 32.0, 96.6, 80.0, 77.0% for 200 U/L, respectively. Area under the curve was 0.663. Because of low sensitivity, routine determination of ADA activity in sputum for the diagnosis of pulmonary tb is not recommended. However, it can be helpful in the diagnosis of smear-negative cases who are strongly suspected of tb.

6319. Eugen-Olsen J, Gustafson P, Sidenius N, Fischer TK, Parner J, Aaby P, Gomes VF, Lisse I. The serum level of soluble urokinase receptor is elevated in tuberculosis patients and predicts mortality during treatment: a community study from Guinea-Bissau. Int J Tuberc Lung Dis. 2002 Aug;6(8):686-92.

OBJECTIVE: To investigate whether the serum level of soluble urokinase plasminogen activator receptor (suPAR) carries prognostic information in individuals infected with Mycobacterium tuberculosis. DESIGN: suPAR was measured by ELISA in 262 individuals at the time of enrolment into a cohort based on suspicion of active tuberculosis and in 101 individuals after 8 months of follow-up. RESULTS: The suPAR levels were elevated in patients with active TB compared to TB-negative individuals (P < 0.001). suPAR levels were highest in patients positive for TB on direct microscopy (n = 84, median suPAR 3.17 ng/ml, P < 0.001), followed by patients negative on direct microscopy but culture positive (n = 35, median suPAR 2.41 ng/ml, P = 0.005) and by patients diagnosed on clinical grounds (n = 63, median suPAR 2.13 ng/ml, P = 0.06) compared to 64 TB-negative individuals (median suPAR 1.73 ng/ml). During the 8-month treatment period, 23 TB cases died. In a multivariate Cox model controlling for HIV status, age, sex, CD4 count and type of TB diagnosis, the mortality increase per ng suPAR was 1.25 (95%CI 1.12-1.40). After treatment, suPAR levels had decreased to the levels of TB-negative individuals. CONCLUSIONS: suPAR levels are elevated in TB patients and associated with mortality. Furthermore, suPAR may be a potential marker of treatment efficacy.

6320. Florio W, Bottai D, Batoni G, Esin S, Pardini M, Maisetta G, Campa M. Identification, Molecular Cloning, and Evaluation of Potential Use of Isocitrate Dehydrogenase II of Mycobacterium bovis BCG in Serodiagnosis of Tuberculosis. Clin Diagn Lab Immunol. 2002 Jul;9(4):846-51.

Diagnosis of tuberculosis is time-consuming and requires infrastructures which are often not available in countries with high incidences of the disease. In the present study, an 82-kDa protein antigen was isolated by affinity chromatography and was identified by peptide mass fingerprinting as isocitrate dehydrogenase II, which is encoded by the icd2 gene of Mycobacterium bovis BCG. The icd2 gene of BCG was cloned by PCR, and the product of recombinant gene expression was purified and analyzed by two-dimensional polyacrylamide gel electrophoresis. The recombinant protein, named rICD2, was tested for its recognition by immunoglobulin G (IgG) antibodies from the sera of 16 patients with tuberculosis (TB) and 23 healthy individuals by Western blotting. The results showed that rICD2 is recognized by IgG antibodies from the sera of all TB patients tested at serum dilutions of >/=1:640. At a serum dilution of 1:1,280, the sensitivity was 50% and the specificity was 86.9%. These results indicate that rICD2 might represent a candidate for use in a new assay for the serodiagnosis of TB.

6321. Gayathri Devi DR; Belwadi S; Beena PM. Department of Microbiology, M.S. Ramaiah Medical College, Bangalore-560002 Diagnostic value of IgA in tuberculosis Indian Practitioner. 2002 Feb; 55(2): 97-9

ABSTRACT: The aim of the study was evolution of the utility of ELISA test using IgA anti-KP-90 antigen for the rapid diagnosis of pulmonory tuberculosis in adults. Thirty one, healthy adult volunteers were considered as subjects for control group. Likewise, twenty clinically suspected cases based on clinical signs and symptoms of tuberculosis were selected for the study. This study showed that sensitivity for diagnosing TB by ZN staining of sputum, culture technique, mantoux test and IgA assay in sera as 100 percent, 65 percent, 40 percent, and 75 percent respectively with positive predictive value of 100 percent of ZN staining of sputum and culture technique, 57 percent for mantoux test, and 74 percent for IgA assay. It is concluded from this study that diagnosing TB by ZN staining of sputum is still the best method.

6322. Handa U, Palta A, Mohan H, Punia RP. Fine needle aspiration diagnosis of tuberculous lymphadenitis. Trop Doct. 2002 Jul;32(3):147-9.

The diagnosis and management of peripheral lymph node tuberculosis remains a major problem in most of the developing countries. We retrieved 584 cases of tuberculous lymphadenitis from a total 1124 lymph node aspirations done over a period of 3 years (1995-1998). Overall acid-fast bacillus positivity was 37.4%, being highest in the cases in which purulent material was aspirated. Fine needle aspiration (FNA) of tuberculous lymphadenopathy provided a high level of diagnostic accuracy as shown by 1.7% false negative and a zero false positive rate. FNA is reliable as an initial evaluating procedure for diagnosis of tuberculous lymphadenitis making it suitable for wider application in developing countries with scant resources.

6323. Houghton RL, Lodes MJ, Dillon DC, Reynolds LD, Day CH, McNeill PD, Hendrickson RC, Skeiky YA, Sampaio DP, Badaro R, Lyashchenko KP, Reed SG. Use of multiepitope polyproteins in serodiagnosis of active tuberculosis. Clin Diagn Lab Immunol. 2002 Jul;9(4):883-91.

Screening of genomic expression libraries from Mycobacterium tuberculosis with sera from tuberculosis (TB) patients or rabbit antiserum to M. tuberculosis led to the identification of novel antigens capable of detecting specific antibodies to M. tuberculosis. Three antigens, Mtb11 (also known as CFP-10), Mtb8, and Mtb48, were tested together with the previously reported 38-kDa protein, in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in TB patients. These four proteins were also produced as a genetically fused polyprotein, which was tested with two additional antigens, DPEP (also known as MPT32) and Mtb81. Sera from individuals with pulmonary and extrapulmonary TB, human immunodeficiency virus (HIV)-TB coinfections, and purified protein derivative (PPD)-positive and PPD-negative status with no evidence of disease were tested. In samples from HIV-negative individuals, the ELISA detected antibodies in >80% of smear-positive individuals and >60% smear-negative individuals, with a specificity of approximately 98%. For this group, smears detected 81.6% but a combination of smear and ELISA had a sensitivity of approximately 93%. The antigen combination detected a significant number of HIV-TB coinfections as well as antibodies in patients with extrapulmonary infections. Improved reactivity in the HIV-TB group was observed by including the antigen Mtb81 that was identified by proteomics. The data indicate that the use of multiple antigens, some of which are in a single polyprotein, can be used to facilitate the development of a highly sensitive test for M. tuberculosis antibody detection.

6324. Hrabec E, Strek M, Zieba M, Kwiatkowska S, Hrabec Z. Circulation level of matrix metalloproteinase-9 is correlated with disease severity in tuberculosis patients. Int J Tuberc Lung Dis. 2002 Aug;6(8):713-9.

SETTING: Parenchymal lung destruction accompanied by active tuberculosis is, at least in part, caused by host as well as bacillus metalloproteinases. Mycobacterium tuberculosis has been shown to stimulate MMP-9 expression in the lung of infected organisms. DESIGN: We have used quantitative zymography and computer-assisted image analysis to measure the levels of type IV collagenases in 20 serum samples of patients with active tuberculosis and in 23 serum samples of healthy volunteers. RESULTS: Mean levels of the serum MMP-9 were over three-fold higher in tuberculous samples compared with normal serum (P < 0.0001), whereas the MMP-2 levels did not differ in these two groups. The levels of MMP-9 were significantly higher in subjects with advanced disease than in those with only limited disease changes (P < 0.05). CONCLUSIONS: We suppose that the elevation of serum MMP-9 levels in patients with tuberculosis is affected by the augmentation of synthesis and/or secretion of this enzyme by inflammatory cells in response to M. tuberculosis infection. The observed association between the serum MMP-9 level and the extent of radiological change suggests that the quantification of the serum level of this enzyme may constitute a supplementary test in pulmonary tuberculosis diagnostics.

6325. Iyer Ranganathan N, Menon MM. Evaluation of varies modalities for the diagnosis of tuberculosis in body fluids. Indian J Path Microbiol. 1999; 42(4): 435-9. No abstract.

6326. Kafwabulula M, Ahmed K, Nagatake T, Gotoh J, Mitarai S, Oizumi K, Zumla A. Evaluation of PCR-based methods for the diagnosis of tuberculosis by identification of mycobacterial DNA in urine samples. Int J Tuberc Lung Dis. 2002 Aug;6(8):732-7.

SETTING: The Chest Clinic and the JICA (Japan International Cooperation Agency) Molecular Laboratories, University Teaching Hospital, Lusaka, Zambia, and the Department of Internal Medicine, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan. OBJECTIVE: To evaluate the polymerase chain reaction (PCR) as a laboratory test for the rapid diagnosis of pulmonary tuberculosis in the African situation by identifying mycobacterial DNA in urine samples using two commonly described molecular methods. DESIGN: Prospective collection and laboratory analysis of urine samples from adult Zambian patients with culture-confirmed pulmonary tuberculosis and healthy controls. METHODS: Urine was obtained from 63 patients with culture-confirmed active pulmonary tuberculosis and 63 'healthy' control patients with no active tuberculosis. DNA was isolated from urine sediment and subjected to analyses by two well-described PCR-based methods, 'the Sechi method' and 'the Githui method', for the identification of Mycobacterium tuberculosis DNA. The sensitivity and specificity of the two tests were determined. RESULTS: The sensitivity and specificity of the Githui method were 55.6% (35/63) and 98.4% (62/63), respectively. The sensitivity and specificity of the Sechi method were 28.6% (18/63) and 98.4% (62/63), respectively. Of the 63 patients, 50 (79%) were HIV sero-positive and the frequency of positive PCR urines using the Githui method was greater in HIV-positive patients than in HIV-negative patients (32/50 = 64% vs. 3/13 = 23%; P = 0.05). CONCLUSIONS: Neither the Githui method nor the Sechi method was sensitive enough to be recommended for routine use in clinical practice. PCR-based assays for the detection of M. tuberculosis DNA in urine will require further refinement before they can be recommended for use in clinical practice in Africa. The presence of mycobacterial DNA in urine samples of patients with pulmonary tuberculosis also requires further study.

6327. Kaminogo M, Ishimaru H, Morikawa M, Suzuki Y, Shibata S. Proton MR spectroscopy and diffusion-weighted MR imaging for the diagnosis of intracranial tuberculomas. Report of two cases. Neurol Res. 2002 Sep;24(6):537-43.

With the current prevalence of tuberculosis, the incidence of intracranial tuberculoma may be on the rise in industrialized nations. However, clinical findings suggestive of systemic tuberculosis are frequently subtle or absent in patients with intracranial tuberculoma, and no specific neuroradiologic characteristics of tuberculoma have been defined as yet. We report two cases of ring-enhanced intracranial tuberculoma in which magnetic resonance (MR) proton spectroscopy and diffusion-weighted (DW) imaging were useful in the differential diagnosis between tuberculoma and other ring-enhanced mass lesions. Pulmonary tuberculosis had been diagnosed in one patient, but radiologic lung study and tuberculin skin test were negative in the other. DW imaging showed bright signal intensity in the core of all lesions in both patients. Malignant gliomas and metastatic brain tumors do not have this characteristic. Proton MR spectroscopy of lesion cores showed lipid peaks and a choline peak in one, and a lipid/lactate mixture pattern in the other, which differed distinctively from those of the pyogenic brain abscess. in each case, one lesion was surgically removed. Antituberculosis drugs were started before surgery for one patient and after surgery for the other. In both, the remaining lesions were reduced significantly in size. We discuss the diagnostic potential of these MR techniques and management options of intracranial tuberculoma.

6328. Kejariwal D, Sarkar N, Chakraborti SK, Agarwal V, Roy S. Pyrexia study of 100 cases. J Postgrad Med 2001; 47 (2): 104-107. No abstract.

6329. Kruuner A, Pehme L, Ghebremichael S, Koivula T, Hoffner SE, Mikelsaar M. Use of molecular techniques to distinguish between treatment failure and exogenous reinfection with Mycobacterium tuberculosis. Clin Infect Dis. 2002 Jul 15;35(2):146-55.

We investigated the means by which drug resistance emerges among drug susceptible Mycobacterium tuberculosis strains during antituberculosis therapy. Patients who experienced failure of treatment for active pulmonary tuberculosis, who initially received diagnoses of infection with drug susceptible M. tuberculosis, and who had had at least 3 isolates tested for drug susceptibility were selected from a 6-year period in the Estonian National Reference Laboratory archive. Eleven patients from whom 35 sequential isolates of M. tuberculosis had been obtained were recruited into the study. Their clinical data and treatment charts were analyzed and correlated with drug-susceptibility patterns and IS6110 restriction fragment-length polymorphism (RFLP) profiles. Six patients excreted isogenic drug-susceptible M. tuberculosis strains, whereas, in the other 5 patients, the isolated strain shifted from a susceptible to a resistant phenotype. In all cases, this shift correlated to a shift in RFLP pattern, which showed reinfection with a new strain. Exogenous reinfection with drug-resistant M. tuberculosis may be misinterpreted as the emergence of drug resistance if molecular testing techniques are not used.

6330. Kumar P, Sharma N, Sharma NC, Patnaik S. Clinical profile of tuberculosis in patients with HIV Infection/AIDS. Indian J Chest Dis Allied Sci. 2002 Jul-Sep;44(3):159-63.

Tuberculosis is said to be one of the commonest opportunistic infection in patients with HIV/ AIDS. A study was carried out to study the clinical, bacteriological and radiological features of HIV/TB patients. Over a period of two years, a total of 301 tuberculosis patients were suspected to have HIV/AIDS co-infection, and upon testing, 42 patients were found to be HIV seropositive. Most of the study patients were manual labourers followed by truck drivers. Sexual (heterosexual) route was found to be the major risk factor for HIV/AIDS. The most common symptom in these patients was cough and expectoration, followed by fever and weight loss. Acid-fast bacilli (AFB) smear positivity was found in 21.4% patients. On chest skiagram, infiltrative lesions were commonly seen in 61.9% patients. Extra-pulmonary tubercular manifestations were seen in 45.6% of HIV/TB cases.

6331. Maheshwari V, Srivastava VK, Prasad S, Alam K. Tuberculoma--A significant diagnostic entity in brain biopsies of intracranial space occupying lesions in children. J Trop Pediatr. 2002 Aug;48(4):242-4. No abstract.

6332. Mathew S, Paramasivan C N. Use of vancomycin in the culture of Mycobacterium tuberculosis from gastric lavage. Indian J med Res 2001; 113(Apr), 125-8. No abstract.

6333. Moonan PK, Quitugua T, Cox RA, Weis SE. Associate investigations: detection of tuberculosis infections in children resulting in discovery of undiagnosed tuberculosis in adults. J Am Osteopath Assoc. 2002 Jul;102(7):397-400. The authors present the design and implementation of associate investigations of young children with positive tuberculin skin test results. Case study analysis of an associate investigation was done using epidemiologic surveillance techniques, medical interviewing, sociogram mapping, tuberculin skin testing, radiographic evidence, and bacteriologic analysis. Deoxyribonucleic acid fingerprinting of the Mycobacterium tuberculosis isolates using a standardized IS6110-based restriction fragment length polymorphism analysis and IS6110-independent DNA spoligotyping methods was done to track and identify specific bacterial strains. Deoxyribonucleic acid fingerprinting and spoligotyping done on isolates obtained from family members demonstrated same-strain transmission of M. tuberculosis. Three adults with active pulmonary disease and six individuals with latent tuberculosis (TB) were discovered during this investigation. The arrival of a family member from Mexico who had the same strain suggests that the source case lives in Mexico. A child with positive tuberculin skin test results indicates recent and potentially ongoing transmission of TB in the community. Targeted tuberculin skin testing performed on high-risk groups by primary care physicians allows for detection of TB infections. When TB infections are discovered in children, associate investigations can result in the discovery of undiagnosed adult cases and prevent further transmission within the community.

6334. Muzaffar R, Batool S, Aziz F, Naqvi A, Rizvi A. Evaluation of the FASTPlaqueTB assay for direct detection of Mycobacterium tuberculosis in sputum specimens. Int J Tuberc Lung Dis. 2002 Jul;6(7):635-40.

SETTING: Sputum samples were collected from suspected tuberculosis patients attending out-patient clinics at the Ojha Institute of Chest Diseases, Karachi, Pakistan. OBJECTIVE: To evaluate the performance of the FASTPlaqueTB assay for rapid diagnosis of pulmonary tuberculosis. DESIGN: A comparative study of 584 sputum samples using acid-fast smear microscopy, Lowenstein-Jensen culture and FASTPlaqueTB. RESULTS: A total of 514 samples yielded complete results. Seventy specimens were lost to analysis due to the overgrowth of contaminants. The addition of antimicrobials inhibited growth of gram-positive contaminants, and reduced the overall contamination rate from 18.2% to 7.2%. Mycobacterium tuberculosis was isolated from 175 smear-positive and 70 smear-negative specimens. FASTPlaqueTB detected M. tuberculosis in 81.6% of specimens, with a specificity of 97.7%. The sensitivity and specificity of the assay for smear-positive specimens were respectively 87.4% and 88.2%. For smear-negative specimens, the sensitivity of the assay was 67.1% and the specificity was 98.4%. The combined sensitivity of smear and FASTPlaqueTB for M. tuberculosis was 90%. Test results were available in 48 hours. CONCLUSION: FASTPlaqueTB is a sensitive and specific test for rapid diagnosis of pulmonary tuberculosis in high prevalence areas. The test is sensitive enough to confirm a large number of clinically suspected smear-negative cases and improve case finding.

6335. Panda D, Lahiri A, Bharracharya I, Ladiri M, Maitra MK. Humoral immune responses in different clinical forms of tuberculosis. J Indian med Ass 2001; 99(8): 424-7. No abstract.

6336. Patzina RA, de Andrade HF Jr, de Brito T, Filho HC, Kauffman MR, Pagliari C, Lucena A, Ribeiro da Matta VL, Seixas Duarte MI. Molecular and standard approaches to the diagnosis of mycobacterial granulomatous lymphadenitis in paraffin-embedded tissue. Lab Invest. 2002 Aug;82(8):1095-7. No abstract.

6337. S. Banerjee, S. Gupta , N. Shende, S. Kumar & B.C. Harinath, Cocktail of ES-31 and ES-41 antigens for screening of pulmonary and extrapulmonary tuberculosis Biomedical Research 2002; 13(2).

6338. Selvakumar N, Gomathi M, Rehman F, Narayanan PR. Evaluation of a two-reagent cold staining method for detection of acid-fast bacilli. Int J Tuberc Lung Dis. 2002 Aug;6(8):728-31.

SETTING: Tuberculosis Research Centre, Chennai, India. OBJECTIVE: To evaluate a two-reagent cold staining method for detection of acid-fast bacilli in sputum smears. SPUTUM SAMPLES: Two hundred and forty-four samples from pulmonary tuberculosis patients attending Tuberculosis Research Centre were used. METHODS AND DESIGN: Two smears were prepared from each of the samples, of which one was allotted to the two-reagent cold staining method and the other to the Ziehl-Neelsen (Z-N) method. The smears were read blind by a single technician. To ensure correct grading, a senior technician checked all positives and 20% of the negative smears. All the samples were processed by modified Petroff's method for culture of Mycobacterium tuberculosis. RESULTS: The concordance (smear grade one above and one below) between the methods was 90% (kappa value, 0.7). The performance of the cold method and the Z-N method was similar when their smear results were compared with culture results (cold method vs. culture, kappa = 0.61; ZN method vs. culture, kappa = 0.67) CONCLUSION: The two-reagent cold staining method was found to be as sensitive and specific as the Z-N method. However, large-scale multicentric studies in different climatic conditions need to be conducted to assess its efficacy in the diagnosis of pulmonary tuberculosis.

6339. Selvakumar N, Rahman F, Garg R, Rajasekaran S, Mohan NS, Thyagarajan K, Sundaram V, Santha T, Frieden TR, Narayanan PR. Evaluation of the phenol ammonium sulfate sedimentation smear microscopy method for diagnosis of pulmonary tuberculosis. J Clin Microbiol. 2002 Aug;40(8):3017-20.

We compared the sensitivity and specificity of the phenol ammonium sulfate (PhAS) sediment smear microscopy method for detection of acid-fast bacilli with those of direct smear microscopy, using culture results for Mycobacterium tuberculosis as the "gold standard." The sensitivities of the PhAS and direct smear methods were 85% (465 of 547) and 83% (454 of 547), respectively, and the specificity of each method was 97%. The PhAS method was better accepted by the laboratory technicians and safer but necessitates an overnight sedimentation, which delays reporting of results until 1 day after sputum collection.

6340. Shenai S, Rodrigues C, Almeida A, Mehta A. Rapid diagnosis of tuberculosis using transcription mediated amplification. Indian J med Microbiol. 2001; 19(4): 184-9. No abstract.

6341. Southey A, Costello E, Gormley E. Detection of Mycobacterium bovis infection and production of interleukin-2 by in vitro stimulation of badger lymphocytes. Vet Immunol Immunopathol. 2002 Aug;87(1-2):73-8.

The Eurasian badger (Meles meles) is considered to be an important wildlife reservoir for Mycobacterium bovis infection of cattle in Ireland and in GB. However, rapid diagnosis of tuberculosis in live badgers has been constrained through a lack of suitable immuno-diagnostic reagents for detection of M. bovis-infected animals. To date, there have been no reports of cytokine activity in badgers that might be associated with specific immune responses to M. bovis infection. In this study, nine badgers were removed from an area with a persistent tuberculosis problem in cattle herds and tuberculosis was confirmed in four of the animals by "post-mortem" examination and M. bovis culture. In preliminary investigations of interleukin-2 (IL-2) activity, we were able to demonstrate that lymphoblasts prepared from badger peripheral blood mononuclear cells (PBMCs) proliferated when cultured in the presence of human recombinant IL-2 (HrIL-2). Supernatants derived from purified protein derivative of tuberculin (PPD-bovine) stimulated PBMC cultures also induced blastogenesis of badger-derived lymphoblasts. The results demonstrate that badger lymphocytes are responsive to HrIL-2 and that PPD-bovine stimulation of badger PBMC results in production of bio-active IL-2.

6342. Sriram U, Selvaraj P, Kurian S M, Reetha A M, Narayanan P R. HLA-DR2 subtypes and immune responses in pulmonary tuberculosis. Indian J med Res 2001, 113(Apr), 117-24. No abstract.

6343. Srivastava L, Srivastava VK. Detection of mycobacterial antigen in circulating immune complexes in patients with chidhood tuberculosis. Indian J Path Microbiol. 1999; 42(4): 405-09. No abstract.

6344. Walravens K, Marche S, Rosseels V, Wellemans V, Boelaert F, Huygen K, Godfroid J. IFN-gamma diagnostic tests in the context of bovine mycobacterial infections in Belgium. Vet Immunol Immunopathol. 2002 Sep 10;87(3-4):401-6.

In countries where cattle tuberculosis caused by Mycobacterium bovis (Mbov) and paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (Mptb) are present, testing strategies for the Mbov eradication have to discriminate between these two infections. Present indirect tests are based on the analysis of the specific cellular immune response (DTH, IFN-gamma) against crude mycobacterial antigens (avian and bovine PPD). In this study, we compared the evolution of the IFN-gamma responses of animals experimentally infected with Mbov, Mptb, or inoculated with Mycobacterium phlei. Mbov inoculation induced a strong IFN-gamma response that allows rapid classification of the status of the animals following interpretation criteria set up by us. Experimental inoculation with M. phlei induced sensitisation to mycobacterial antigens as detected by the IFN-gamma test but these reactions were of short duration, therefore, repeated testing allows us to define these animals as aspecific reactors. IFN-gamma response induced after oral inoculation of calves with Mptb was of low intensity and ratio of responses measured against avian versus bovine PPD did not allow a clear diagnostic at least for the six first month of infection.

6345. Yilmaz T, Sever A, Gur S, Killi R, Elmas N. CT findings of abdominal tuberculosis in 12 patients. Comput Med Imaging Graph. 2002 Sep-Oct;26(5):321.

Our purpose was to evaluate the Computed Tomography (CT) findings of the abdominal tuberculosis (TBC) retrospectively which was diagnosed histopatologically. This study included 12 patients. All patients were evaluated by abdominal CT study. Most findings of CT studies were mesenteric calcified or noncalcified lymphadenopathies, ascites, thickened intestinal wall located on the right lower quadrant of abdomen, thickening of peritoneum, mottled soft-tissue densities in omentum and mesenterium. In addition, one of the patients had bilateral calcified adrenal glands and one of them had calcified mass in adrenal gland. If peritoneal thickening, ascites, abdominal lymphadenophaties and thickened intestinal walls are obtained, TBC should be considered in differential diagnosis in developing countries.

Pahtogenesis:

6346. Ameixa C, Friedland JS. Interleukin-8 secretion from Mycobacterium tuberculosis-infected monocytes is regulated by protein tyrosine kinases but not by ERK1/2 or p38 mitogen-activated protein kinases. Infect Immun. 2002 Aug;70(8):4743-6.

Mycobacterium tuberculosis upregulates NF-kappaB binding and interleukin-8 (IL-8) gene expression and secretion in primary human monocytes. Inhibition of tyrosine protein kinases but not of ERK1/2 or p38 mitogen-activated protein kinases downregulates tuberculosis-induced IL-8 secretion. The inhibitor genistein decreased NF-kappaB nuclear translocation and IL-8 gene transcription in addition to acting on posttranscriptional processing.

6347. Boechat N, Lagier-Roger B, Petit S, Bordat Y, Rauzier J, Hance AJ, Gicquel B, Reyrat JM. Disruption of the gene homologous to mammalian Nramp1 in Mycobacterium tuberculosis does not affect virulence in mice. Infect Immun. 2002 Aug;70(8):4124-31.

Natural-resistance-associated macrophage protein 1 (Nramp1) is a divalent cation transporter belonging to a family of transporter proteins highly conserved in eukaryotes and prokaryotes. Mammalian and bacterial transporters may compete for essential metal ions during mycobacterial infections. The mycobacterial Nramp homolog may therefore be involved in Mycobacterium tuberculosis virulence. Here, we investigated this possibility by inactivating the M. tuberculosis Nramp1 gene (Mramp) by allelic exchange mutagenesis. Disruption of Mramp did not affect the extracellular growth of bacteria under standard conditions. However, the Mramp mutation was associated with growth impairment under conditions of limited iron availability. The Mramp mutant displayed no impairment of growth or survival in macrophages derived from mouse bone marrow or in Nramp1(+/+) and Nramp1(-/-) congenic murine macrophage cell lines. Following intravenous challenge in BALB/c mice, counts of parental and Mramp mutant strains were similar in the lungs and spleens of the animals at all time points studied. These results indicate that Mramp does not contribute to the virulence of M. tuberculosis in mice.

6348. Chackerian AA, Alt JM, Perera TV, Dascher CC, Behar SM. Dissemination of Mycobacterium tuberculosis is influenced by host factors and precedes the initiation of T-cell immunity. Infect Immun. 2002 Aug;70(8):4501-9.

We report that dissemination of Mycobacterium tuberculosis in the mouse is under host control and precedes the initiation of T-cell immunity. Nine to eleven days after aerosol inoculation, M. tuberculosis disseminates to the pulmonary lymph nodes (LN), where M. tuberculosis-specific T cells are detected 2 to 3 days thereafter. This indicates that the initial spread of bacteria occurs via lymphatic drainage and that the acquired T-cell immune response is generated in the draining LN. Dissemination to peripheral sites, such as the spleen and the liver, occurs 11 to 14 days postinfection and is followed by the appearance of M. tuberculosis-specific T cells in the lung and the spleen. In all cases studied, dissemination to the LN or the spleen preceded activation of M. tuberculosis-specific T cells in that organ. Interestingly, bacteria disseminate earlier from the lungs of resistant C57BL/6 mice than from the lungs of susceptible C3H mice, and consequently, C57BL/6 mice generate an immune response to M. tuberculosis sooner than C3H mice generate an immune response. Thus, instead of spreading infection, early dissemination of M. tuberculosis may aid in the initiation of an appropriate and timely immune response. We hypothesize that this early initiation of immunity following inoculation with M. tuberculosis may contribute to the superior resistance of C57BL/6 mice.

6349. Fisher MA, Plikaytis BB, Shinnick TM. Microarray analysis of the Mycobacterium tuberculosis transcriptional response to the acidic conditions found in phagosomes. J Bacteriol. 2002 Jul;184(14):4025-32.

We used microarrays and real-time reverse transcription-PCR to analyze the global transcriptional response of Mycobacterium tuberculosis to low pH in vitro, which may mimic an environmental signal encountered by phagocytosed mycobacteria. Eighty-one genes were differentially expressed >1.5-fold, including many involved in fatty acid metabolism. The most highly induced genes showed homology with nonribosomal peptide synthetases/polyketide synthases.

6350. Garton NJ, Gilleron M, Brando T, Dan HH, Giguere S, Puzo G, Prescott JF, Sutcliffe IC. A novel lipoarabinomannan from the equine pathogen Rhodococcus equi. Structure and effect on macrophage cytokine production. J Biol Chem. 2002 Aug 30;277(35):31722-33.

Rhodococcus equi is a major cause of foal morbidity and mortality. We have investigated the presence of lipoglycan in this organism as closely related bacteria, notably Mycobacterium tuberculosis, produce lipoarabinomannans (LAM) that may play multiple roles as virulence determinants. The lipoglycan was structurally characterized by gas chromatography-mass spectrometry following permethylation, capillary electrophoresis after chemical degradation, and (1)H and (31)P and two-dimensional heteronuclear nuclear magnetic resonance studies. Key structural features of the lipoglycan are a linear alpha-1,6-mannan with side chains containing one 2-linked alpha-d-Manp residue. This polysaccharidic backbone is linked to a phosphatidylinositol mannosyl anchor. In contrast to mycobacterial LAM, there are no extensive arabinan domains but single terminal alpha-d-Araf residue capping the 2-linked alpha-d-Manp. The lipoglycan binds concanavalin A and mannose-binding protein consistent with the presence of t-alpha-d-Manp residues. We studied the ability of the lipoglycans to induce cytokines from equine macrophages, in comparison to whole cells of R. equi. These data revealed patterns of cytokine mRNA induction that suggest that the lipoglycan is involved in much of the early macrophage cytokine response to R. equi infection. These studies identify a novel LAM variant that may contribute to the pathogenesis of disease caused by R. equi.

6351. Heldwein KA, Fenton MJ. The role of Toll-like receptors in immunity against mycobacterial infection. Microbes Infect. 2002 Jul;4(9):937-44. Review.

Recent work implicates Toll-like receptor (TLR) proteins as regulators of innate immune cell activation induced by Mycobacterium tuberculosis, which continues to ravage nearly one-third of the world's population. Novel insights into how TLR proteins may dictate the nature and extent of cellular immune responses against this pathogen will be discussed.

6352. Johansen IS, Lundgren BH, Thyssen JP, Thomsen VO. Rapid differentiation between clinically relevant mycobacteria in microscopy positive clinical specimens and mycobacterial isolates by line probe assay. Diagn Microbiol Infect Dis. 2002 Aug;43(4):297-302.

The Inno LiPA Mycobacteria assay, based on PCR amplification of the 16-23S rRNA spacer region of Mycobacterium species, has been designed for identification of mycobacteria grown in culture media and discrimination between Mycobacterium tuberculosis complex, M. avium, M. intracellulare, M. kansasii, M. gordonae, M. xenopi, scrofulaceum and M. chelonae group including M. abscessus. In order to evaluate the system as a fast diagnostic tool, the assay was for the first time used directly on 14 microscopy positive clinical specimens and 71 isolates and the results were compared to those of conventional identification using 16S rDNA analysis and biochemical properties. The assay only misidentified one strain, which was found to be M. avium complex instead of M. intracellulare as found by the conventional tests. The assay allows rapid discrimination of the eight most clinically relevant mycobacteria in microscopy positive clinical specimens and isolates within 6 h in the same procedural run.

6353. Kinjo Y, Kawakami K, Uezu K, Yara S, Miyagi K, Koguchi Y, Hoshino T, Okamoto M, Kawase Y, Yokota K, Yoshino K, Takeda K, Akira S, Saito A. Contribution of IL-18 to Th1 response and host defense against infection by Mycobacterium tuberculosis: a comparative study with IL-12p40. J Immunol. 2002 Jul 1;169(1):323-9.

The present study was conducted to critically determine the protective role of IL-18 in host response to Mycobacterium tuberculosis infection. IL-18-deficient (knockout (KO)) mice were slightly more prone to this infection than wild-type (WT) mice. Sensitivity of IL-12p40KO mice was lower than that of IL-12p40/IL-18 double KO mice. IFN-gamma production caused by the infection was significantly attenuated in IL-18KO mice compared with WT mice, as indicated by reduction in the levels of this cytokine in sera, spleen, lung, and liver, and its synthesis by spleen cells restimulated with purified protein derivatives. Serum IL-12p40 level postinfection and its production by peritoneal exudate cells stimulated with live bacilli were also significantly lower in IL-18KO mice than WT mice, suggesting that attenuated production of IFN-gamma was secondary to reduction of IL-12 synthesis. However, this was not likely the case, because administration of excess IL-12 did not restore the reduced IFN-gamma production in IL-18KO mice. In further studies, IL-18 transgenic mice were more resistant to the infection than control littermate mice, and serum IFN-gamma level and its production by restimulated spleen cells were increased in the former mice. Taken together, our results indicate that IL-18 plays an important role in Th1 response and host defense against M. tuberculosis infection although the contribution was not as profound as that of IL-12p40.

6354. Kisich KO, Higgins M, Diamond G, Heifets L. Tumor necrosis factor alpha stimulates killing of Mycobacterium tuberculosis by human neutrophils. Infect Immun. 2002 Aug;70(8):4591-9.

The ability of human neutrophils to aid in defense against pulmonary infection with Mycobacterium tuberculosis is controversial. In this study, we have shown that neutrophils respond to and phagocytose M. tuberculosis in human lesions. Neutrophils from healthy individuals were able to kill significant fractions of an inoculum of M. tuberculosis within 1 h of phagocytosis, and this ability was enhanced by tumor necrosis factor alpha but not by gamma interferon. The mycobactericidal mechanism was nonoxidative, as inhibitors of reactive oxygen or reactive nitrogen intermediates did not interfere with killing. However, the mycobactericidal mechanism was associated with increased exposure of intracellular M. tuberculosis to neutrophil defensins. In vitro, human neutrophil peptides 1 to 3 were not able to kill the bacilli even at much higher levels. These studies support the concept that human neutrophils are directly involved in defense against infection with M. tuberculosis.

6355. Mokrousov I, Otten T, Filipenko M, Vyazovaya A, Chrapov E, Limeschenko E, Steklova L, Vyshnevskiy B, Narvskaya O. Detection of isoniazid-resistant Mycobacterium tuberculosis strains by a multiplex allele-specific PCR assay targeting katG codon 315 variation. J Clin Microbiol. 2002 Jul;40(7):2509-12.

We describe a simple multiplex allele-specific (MAS)-PCR assay to detect mutations in the second base of the katG gene codon 315, including AGC- >ACC and ACA (Ser-->Thr) substitutions that confer resistance to isoniazid (INH) in Mycobacterium tuberculosis clinical isolates. The 315 ACC allele is found in the majority of Inh(r) strains worldwide, especially in areas with a high incidence of tuberculosis. The 315 ACA allele is characteristic of the New York City multidrug-resistant (MDR) strain W and its progenies in the United States. The mutations in katG315 are revealed depending on the presence or absence of an indicative fragment amplified from the wild-type allele of this codon. Initially optimized on the purified DNA samples, the assay was then tested on crude cell lysates and auramine-stained sputum slide preparations with the same reproducibility and interpretability of profiles generated by agarose gel electrophoresis. The MAS-PCR assay can be used for the detection of resistance to INH in clinical laboratories in regions with a high prevalence of MDR M. tuberculosis strains.

6356. Nathan C. Inducible nitric oxide synthase in the tuberculous human lung. Am J Respir Crit Care Med. 2002 Jul 15;166(2):130-1. No abstract.

6357. Paine K, Flower DR. Bacterial bioinformatics: pathogenesis and the genome. J Mol Microbiol Biotechnol. 2002 Jul;4(4):357-65.

As the number of completed microbial genome sequences continues to grow, there is a pressing need for the exploitation of this wealth of data through a synergistic interaction between the well-established science of bacteriology and the emergent discipline of bioinformatics. Antibiotic resistance and pathogenicity in virulent bacteria has become an increasing problem, with even the strongest drugs useless against some species, such as multi-drug resistant Enterococcus faecium and Mycobacterium tuberculosis. The global spread of Human Immunodeficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS) has contributed to the re-emergence of tuberculosis and the threat from new and emergent diseases. To address these problems, bacterial pathogenicity requires redefinition as Koch's postulates become obsolete. This review discusses how the use of bacterial genomic information, and the in silico tools available at present, may aid in determining the definition of a current pathogen. The combination of both fields should provide a rapid and efficient way of assisting in the future development of antimicrobial therapies.

6358. Pathania R, Navani NK, Gardner AM, Gardner PR, Dikshit KL. Nitric oxide scavenging and detoxification by the Mycobacterium tuberculosis haemoglobin, HbN in Escherichia coli. Mol Microbiol. 2002 Sep;45(5):1303-14.

Nitric oxide (NO), generated in large amounts within the macrophages, controls and restricts the growth of internalized human pathogen, Mycobacterium tuberculosis H37Rv. The molecular mechanism by which tubercle bacilli survive within macrophages is currently of intense interest. In this work, we have demonstrated that dimeric haemoglobin, HbN, from M. tuberculosis exhibits distinct nitric oxide dioxygenase (NOD) activity and protects growth and cellular respiration of heterologous hosts, Escherichia coli and Mycobacterium smegmatis, from the toxic effect of exogenous NO and the NO-releasing compounds. A flavohaemoglobin (HMP)-deficient mutant of E. coli, unable to metabolize NO, acquired an oxygen-dependent NO consumption activity in the presence of HbN. On the basis of cellular haem content, the specific NOD activity of HbN was nearly 35-fold higher than the single-domain Vitreoscilla haemoglobin (VHb) but was sevenfold lower than the two-domain flavohaemoglobin. HbN-dependent NO consumption was sustained with repeated addition of NO, demonstrating that HbN is catalytically reduced within E. coli. Aerobic growth and respiration of a flavohaemoglobin (HMP) mutant of E. coli was inhibited in the presence of exogenous NO but remained insensitive to NO inhibition when these cells produced HbN, VHb or flavohaemoglobin. M. smegmatis, carrying a native HbN very similar to M. tuberculosis HbN, exhibited a 7.5-fold increase in NO uptake when exposed to gaseous NO, suggesting NO-induced NOD activity in these cells. In addition, expression of plasmid-encoded HbN of M. tuberculosis in M. smegmatis resulted in 100-fold higher NO consumption activity than the isogenic control cells. These results provide strong experimental evidence in support of NO scavenging and detoxification function for the M. tuberculosis HbN. The catalytic NO scavenging by HbN may be highly advantageous for the survival of tubercle bacilli during infection and pathogenesis.

6359. Repique CJ, Li A, Collins FM, Morris SL. DNA immunization in a mouse model of latent tuberculosis: effect of DNA vaccination on reactivation of disease and on reinfection with a secondary challenge. Infect Immun. 2002 Jul;70(7):3318-23.

Individuals who are latently infected with Mycobacterium tuberculosis can develop active disease via either endogenous reactivation of the latent bacilli or exogenous reinfection with a second mycobacterial strain. In this study, we investigated whether immunization with a tuberculosis DNA vaccine cocktail that induces significant protective responses in mice could prevent reactivation of disease in a murine latent-tuberculosis model. In addition, we assessed whether DNA vaccination could retard the growth of a secondary aerogenic infection with M. tuberculosis (exogenous reinfection) in latently infected mice. In the reactivation studies, administration of the DNA vaccine combination did not prevent recrudescence of the latent infection after injection of dexamethasone. Moreover, for the reinfection experiments, only a modest decrease in the growth of a secondary M. tuberculosis challenge in DNA-vaccinated animals, compared to controls, was observed 14 and 28 days after the reinfection of previously exposed mice. Interestingly, although proliferation of the secondary challenge was reduced significantly in a nonvaccinated chronic-infection group relative to the naive controls, the number of bacilli still increased by 500-fold 1 month after the secondary challenge in mice with active tuberculosis. These results indicate that novel immunotherapeutic approaches will be required to prevent reactivation of infection or reinfection of individuals with latent tuberculosis.

6360. Sanchez-Rodriguez C, Estrada-Chavez C, Garcia-Vigil J, Laredo-Sanchez F, Halabe-Cherem J, Pereira-Suarez A, Mancilla R. An IgG antibody response to the antigen 85 complex is associated with good outcome in Mexican Totonaca Indians with pulmonary tuberculosis. Int J Tuberc Lung Dis. 2002 Aug;6(8):706-12.

SETTING: It is generally accepted that antibodies do not protect against Mycobacterium tuberculosis infection, as this role relies upon T-cell reactivity. Hence, most studies on antimycobacterial antibodies have been aimed at developing serologic tests, and few explore their role in disease pathogenesis. OBJECTIVE: To determine the IgG antimycobacterial antibody response of 55 Mexican Totonaca Indians with pulmonary tuberculosis and its correlation with some features of the disease. DESIGN: Study of the profile of antigen recognition by immunoblot and ELISA with isolated antigen 85 complex (Ag85) and whole culture filtrate proteins. Correlation of immunoblot and ELISA results with BCG vaccination, tuberculin reactivity, extent of the disease, clinical setting, and response to treatment. RESULTS: On immunoblot, band reactivity was very poor and the most frequently recognized antigen was the 30-32 kDa, antigen 85 complex (45.8% of serum samples). ELISA with this antigen showed a sensitivity of 72% and a specificity of 100%. Positive antibody titers to Ag85 were observed in 79.4% of patients with non-cavitary tuberculosis (P = 0.012) and in 95.8% of patients who were cured with anti-tuberculosis chemotherapy (P = 0.0001). By contrast, an antibody response to whole culture filtrate antigens had no correlation with the presence of cavitations or with prognosis. CONCLUSIONS: Our data show that an antibody response to Ag85, aside from having great potential to develop a serologic test for tuberculosis, was associated with a positive outcome in a cohort of tuberculous Mexican Indians.

6361. Soborg C, Andersen AB, Madsen HO, Kok-Jensen A, Skinhoj P, Garred P. Natural resistance-associated macrophage protein 1 polymorphisms are associated with microscopy-positive tuberculosis. J Infect Dis. 2002 Aug 15;186(4):517-21.

The natural resistance-associated macrophage protein 1 (NRAMP1) is implicated in the pathophysiology of mycobacterial infections. We investigated by polymerase chain reaction previously published Nramp1 genotypes at 4 loci-INT4, N543D, 3'UTR, and 5'(CA)(n) microsatellite markers-in 104 human immunodeficiency virus-negative patients with tuberculosis and 176 healthy control subjects living in Denmark. No significant difference in genotype frequency was found between white patients with tuberculosis and control subjects (P>.16), but carriage of Nramp1 variant alleles at loci INT4 and 5'(CA)(n) conferred a significantly increased risk of having microscopy-positive compared with microscopy-negative tuberculosis (65% vs. 35% [P=.0004] and 63% vs. 38% [P=.047], respectively). The Nramp1 alleles were not associated with increased risk for the development of cavities seen on chest radiographs, or with extrapulmonary tuberculosis. These results indicate that variant alleles in the Nramp1 gene are associated with increased mycobacterial replication rather than susceptibility for tuberculosis and may thus confer increased risk of severe disease.

6362. Verreck FA, de Boer T, Hoeve M, van Dissel JT, Sanal O, Kurimoto M, Ottenhoff TH. Human host defense and cytokines in mycobacterial infectious diseases: interleukin-18 cannot compensate for genetic defects in the interleukin-12 system. Clin Infect Dis. 2002 Jul 15;35(2):210-2. No abstract.

6363. von Reyn CF, Vuola JM. New vaccines for the prevention of tuberculosis. Clin Infect Dis. 2002 Aug 15;35(4):465-74.

Mycobacterium bovis, bacille Calmette-Guerin (BCG) is administered widely to newborns throughout the world and has been shown to be effective in preventing childhood tuberculosis but not reactivation pulmonary disease or human immunodeficiency virus-associated tuberculosis. Development of a more effective, better standardized, affordable vaccine with durable activity and fewer side effects is a major priority. Contemporary molecular techniques have identified promising immunodominant antigens and novel immunization strategies. Vaccine development has also been informed by an improved understanding of the role of nontuberculous mycobacteria in the efficacy of BCG and in the prevention of tuberculosis. Vaccines under investigation include attenuated or enhanced whole-cell live, whole-cell inactivated, subunit, DNA, and prime-boost vaccines. Several candidate vaccines have demonstrated activity in animal models that is equal to or superior to that of BCG, and human trials are under way. Because there is no identified surrogate marker for protection, identification of an improved vaccine will require long-term efficacy trials in humans.

Vaccines:

6364. Cheon SH, Kampmann B, Hise AG, Phillips M, Song HY, Landen K, Li Q, Larkin R, Ellner JJ, Silver RF, Hoft DF, Wallis RS. Bactericidal activity in whole blood as a potential surrogate marker of immunity after vaccination against tuberculosis. Clin Diagn Lab Immunol. 2002 Jul;9(4):901-7.

The development of new tuberculosis (TB) vaccines will require the identification of correlates of human protection. This study examined the balance between immunity and virulence in a whole blood infection model in which intracellular mycobacterial survival was measured using BACTEC. In the blood of tuberculin-negative donors, counts of Mycobacterium tuberculosis H(37)Ra organisms fell by 0.14 log(10) CFU during 96 h of whole blood culture, whereas counts of Mycobacterium bovis BCG, M. tuberculosis H(37)Rv, and a clinical TB isolate's organisms increased by 0.13, 0.43, and 1.04 log(10) CFU, respectively (P < 0.001), consistent with their relative virulence. Inhibition of tumor necrosis factor alpha by the addition of methylprednisolone or pentoxifylline or removal of CD4(+) or CD8(+) T cells by magnetic beads had deleterious effects on immune control of intracellular growth only in the blood of tuberculin-positive donors. Repeated vaccination of eight tuberculin-negative volunteers with M. bovis BCG resulted in a 0.3 log (50%) reduction in BCG CFU counts in the model compared to baseline values (P < 0.05). Three of the volunteers responded only after the second vaccination. These experiments indicate that whole blood culture may be used to measure immunity to M. tuberculosis and that further studies of repeated BCG vaccination are warranted.

6365. D'Souza S, Rosseels V, Denis O, Tanghe A, De Smet N, Jurion F, Palfliet K, Castiglioni N, Vanonckelen A, Wheeler C, Huygen K. Improved tuberculosis DNA vaccines by formulation in cationic lipids. Infect Immun. 2002 Jul;70(7):3681-8.

Mice were vaccinated with plasmid DNA (pDNA) encoding antigen 85A (Ag85A), Ag85B, or PstS-3 from Mycobacterium tuberculosis either in saline or formulated for intramuscular injections in VC1052:DpyPE (aminopropyl-dimethyl-myristoleyloxy-propanaminium bromide-diphytanoylphosphatidyl-ethanolamine) (Vaxfectin; Vical, Inc., San Diego, Calif.) or for intranasal instillations in GAP-DLRIE:DOPE (aminopropyl-dimethyl-bis-dodecyloxy-propanaminium bromide-dioleoylphosphatidyl-ethanolamine). These two novel cationic and neutral colipid formulations were previously reported to be effective adjuvants for pDNA-induced antibody responses. The levels of Ag85-specific total immunoglobulin G (IgG) and IgG isotypes were all increased 3- to 10-fold by formulation of pDNA in Vaxfectin. The level of production of splenic T-cell-derived Th1-type cytokines (interleukin-2 and gamma interferon) in response to purified Ag85 and to synthetic peptides spanning the entire Ag85A protein was also significantly higher in animals vaccinated with pDNA formulated in Vaxfectin. Cytolytic T-lymphocyte responses generated by pDNA encoding phosphate-binding protein PstS-3 in Vaxfectin were better sustained over time than were those generated by PstS-3 DNA in saline. Intranasal immunization with Ag85A DNA in saline was completely ineffective, whereas administration in GAP-DLRIE:DOPE induced a positive Th1-type cytokine response; however, the extent of the latter response was clearly lower than that obtained following intramuscular immunization with the same DNA dose. Combined intramuscular and intranasal administrations in cationic lipids resulted in stronger immune responses in the spleen and, more importantly, in the lungs as well. Finally, formulation in Vaxfectin increased the protective efficacy of the Ag85B DNA vaccine, as measured by reduced relative light unit counts and CFU counts in the spleen and lungs from mice challenged with bioluminescent M. tuberculosis H37Rv. These results may be of importance for future clinical use of DNA vaccines in humans.

6366. Lalvani A. CD8 cytotoxic T cells and the development of new tuberculosis vaccines. Am J Respir Crit Care Med. 2002 Sep 15;166(6):789-90. No abstract.

Therapy:

6367. Abel B, Thieblemont N, Quesniaux VJ, Brown N, Mpagi J, Miyake K, Bihl F, Ryffel B. Toll-like receptor 4 expression is required to control chronic Mycobacterium tuberculosis infection in mice. J Immunol. 2002 Sep 15;169(6):3155-62.

Endotoxin from Gram-negative bacteria bound to CD14 signals through Toll-like receptor (TLR) 4, while components of Gram-positive bacteria, fungi, and Mycobacterium tuberculosis (M.tb.) preferentially use TLR2 signaling. We asked whether TLR4 plays any role in host resistance to M.tb. infection in vivo. Therefore, we infected the TLR4 mutant C3H/HeJ mice and their controls, C3H/HeN mice, with M.tb. by aerosol. TLR4 mutant mice had a reduced capacity to eliminate mycobacteria from the lungs, spread the infection to spleen and liver, with 10-100 times higher CFU organ levels than the wild-type mice and succumbed within 5-7 mo, whereas most of the wild-type mice controlled infection and survived the duration of the experiment. The lungs of TLR4 mutant mice showed chronic pneumonia with increased neutrophil infiltration, reduced macrophages recruitment, and abundant acid-fast bacilli. Furthermore, the pulmonary expression of TNF-alpha, IL-12p40, and monocyte chemoattractant protein 1 was significantly lower in C3H/HeJ mice when compared with the wild-type controls. C3H/HeJ-derived macrophages infected in vitro with M.tb. produced lower levels of TNF-alpha. Finally, the purified mycobacterial glycolipid, phosphatidylinositol mannosides, induced signaling in both a TLR2- and TLR4-dependent manner, thus suggesting that recognition of phosphatidylinositol mannosides in vivo may influence the development of protective immunity. In summary, macrophage recruitment and the proinflammatory response to M.tb. are impaired in TLR4 mutant mice, resulting in chronic infection with impaired elimination of mycobacteria. Therefore, TLR4 signaling is required to mount a protective response during chronic M.tb. infection.

6368. Sanklecha M. B.C.G. and T.T. Indian Pediatr. 2002 Sep;39(9):880-1; discussion 881. No abstract.

6369. Shiffman J, Beer T, Wu Y. The emergence of global disease control priorities. Health Policy Plan. 2002 Sep;17(3):225-34.

How do global disease control priorities emerge? This paper examines the post-World War II histories of efforts to control three diseases--polio, malaria and tuberculosis--to investigate this issue. The paper draws from the policy studies literature to evaluate three models of the priority generation process. A rational model suggests logical selection based on global burden and the availability of cost-effective interventions. An incremental model suggests a drawn out process in which health priorities emerge gradually and interventions reach affected populations through slow diffusion. A punctuated equilibrium model suggests a more complex pattern: long periods of stability during which interventions are available only to select populations, punctuated by bursts of attention as these interventions spread across the globe in concentrated periods of time. The paper finds that the punctuated equilibrium model corresponds most closely to efforts to control these three diseases. Bursts are associated with the convergence of three conditions: the widespread acceptance of the disease as a threat; a perception that human interventions can control disease transmission; and the formation of a transnational coalition of health actors concerned with fighting the disease. The generation of each condition requires considerable groundwork, the reason for long periods of stability. Initiatives take off rapidly when the conditions couple, the reason for bursts. The paper aims to spark additional research on the subject of global disease control agenda setting, a neglected issue in the health policy literature.

6370. Singh N, Wagener MM, Gayowski T. Safety and efficacy of isoniazid chemoprophylaxis administered during liver transplant candidacy for the prevention of posttransplant tuberculosis. Transplantation. 2002 Sep 27;74(6):892-5.

BACKGROUND: Optimal timing of initiation of isoniazid chemoprophylaxis in liver transplant recipients who test positive on the tuberculin skin test has not been defined. We sought to determine whether isoniazid prophylaxis administered during liver transplant candidacy was safe and effective for the prevention of posttransplantation tuberculosis. METHODS: During a 9-year period, 18 liver transplant candidates with tuberculin skin test greater than 5 mm or recent conversion to positive tuberculin skin test were identified and received isoniazid chemoprophylaxis for 12 months. For each case, a control matched with the patient for underlying liver disease and age (within 5 years of the case) was included. Liver function tests were assessed monthly. The median follow-up was 55 months and ranged up to 107 months for the cases. RESULTS: At baseline, the cases had a total bilirubin of 2.2 mg/dL, alanine aminotransferase of 106 IU/L, prothrombin time of 14.2 sec, and serum albumin of 2.9 gm/dL (mean values). Hepatic function tests did not differ significantly for the cases at 3, 6, 9, and 12 months when compared with those at baseline or between the cases and controls at each of the above time points. Discontinuation of prophylaxis was not required in any of the patients. The outcome (proportion of patients who underwent transplantation or were dead or alive at the last follow-up) and survival time for the cases did not differ significantly from those of the controls (P >0.20). CONCLUSION: In liver transplant candidates at risk for infection after transplantation, isoniazid chemoprophylaxis used during candidacy was well tolerated and did not adversely effect hepatic function or outcome as compared with the control patients.

6371. Tuberculosis Research Centre. Shortening short course chemotherapy: a randomised clinical trial for treatment of smear positive pulmonary tuberculosis with regimens using ofloxacin in the intensive phase Indian Journal of Tuberculosis. 2002 Jan; 49(1): 27-38. No abstract.

April 2003

6903. Agrawala S Abdominal Tuberculosis .Hosp Today 2002 ,7(2),65-70. (014349, Vol 38 No14 ,16 July 2002)

6904. Agrawala S. Childhood Tuberculosis: surgical aspects . Hosp Today 2002 ,7(2),81-7. (014350, Vol 38 No14 ,16 July 2002).

6905. Ariyurek MO, Karcaaltincaba M, Demirkazik FB, Akay H, Gedikoglu G, Emri S. Bilateral multiple pulmonary tuberculous nodules mimicking metastatic disease. Eur J Radiol. 2002 Oct;44(1):33-6.

We present CT findings of a young woman who has bilateral pulmonary nodules mimicking metastases. Clinical presentation with active multiple pulmonary macronodules without cavitation responsive to treatment is an atypical manifestation of pulmonary tuberculosis. We reviewed the causes of multiple pulmonary nodules, role of radiological findings in differential diagnosis and parenchymal manifestations of pulmonary tuberculosis in this report.

6906. Asnis DS, Cherian S, Sun T, Shrestha S, Santucci T Jr.Pulmonary tuberculosis presenting as community-acquired pneumonia. Clin Infect Dis. 2002 Dec 15;35(12):1574-5. No abstract.

6907. Barrett A, Magee JG, Freeman R. An evaluation of the BD ProbeTec ET system for the direct detection of Mycobacterium tuberculosis in respiratory samples. J Med Microbiol. 2002 Oct;51(10):895-8.

In controlling the spread of tuberculosis, early detection of disease caused by organisms of the Mycobacterium tuberculosis complex (MTBC) is vital. The BD ProbeTec ET system provides a method for the direct detection of MTBC by strand displacement amplification. Two hundred and five respiratory samples from patients with a high probability of tuberculosis were assessed by ProbeTec and by microscopy and culture for mycobacteria. ProbeTec positive results were obtained with 101 of 109 samples from which MTBC organisms were isolated. ProbeTec correctly signalled 78 of 81 samples that gave growths of mycobacteria other than tubercle bacilli (MOTT) as negative. Three samples gave false-positive results, corrected on repeat testing. Positive and negative predictive values (PPV, NPV) were 0.97 and 0.90 and the system showed a sensitivity and specificity of 92.7% and 96.0%, respectively. These values rose to PPV 0.97, NPV 0.96, sensitivity 97.1% and specificity 96.0% when data from the small number of gastric lavage samples tested were removed from the analysis. The BD ProbeTec ET system offers a robust and reliable molecular biological approach to the detection of MTBC organisms in respiratory samples in a semi-automated format.

6908. Beg M,Singh M,Azhari S, Bhargav R ,Ali A .Comparative analysis of immunological diagnostic test of mycobactorium tuberculosis .Indian med Gaz 2001 ,135(7),210-13 (ISA 013301, Vol 38 No13 ,1 July 2002)

6909. Biswas D; Agarwal M; Ali A; Bhargava R.Antigen ES-31 : observation from use of antigen, antibody and immune complexed antigen in sero-diagnosis of tuberculosis Indian Journal of Tuberculosis. 2002 Jul; 49(3): 129-31

ABSTRACT: A group of 40 patients suffering from pulmonary tuberculosis comprising adults and children, bacteriologically positive and negative cases was examined for the presence in their sera of anti ES-31 IgG antibody and free and immune complexed ES-31 antigen by stick penicillinase ELISA test. The control population comprised 25 patients suffering from non-tuberculosis pulmonary diseases and 15 healthy individuals. The cut-off titer for antibody detection was 1:600 and for free and immune complexed antigen, 1:300. Using the ES-31 antigen, antibody could be detected in 30 of 40 patients (75 Percent), while among the bacteriologically negative adult patients, antibody could be detected in 84.6 percent of the cases. Using affinity purified anti ES-31 antibody, free antigen could be detected in 25 of the 40 cases (62.5 percent) whereas complexed antigen could be detected in 20 of them (50 percent). Respective specificity values obtained with free, immune complexed antigen and antibody detection tests were 85 percent, 95 percent and 80 percent

6910. Bose M. Genomics and tuberculosis. Indian J Chest Dis Allied Sci. 2002 Oct-Dec;44(4):221-3. No abstract.

6911. Deivanayagam CN, Rajasekaran S, Venkatesan R, Mahilmaran A, Ahmed PR, Annadurai S, Kumar S, Chandrasekar C, Ravichandran N, Pencillaiah R. Prevalence of acquired MDR-TB and HIV co-infection. Indian J Chest Dis Allied Sci. 2002 Oct-Dec;44(4):237-42.

BACKGROUND: Mounting prevalence of primary and acquired multidrug-resistant tuberculosis in India is a sorry reminder of all round failure in our fight against tuberculosis and also of the necessity for new effective strategies. OBJECTIVES: (1) To assess the prevalence and pattern of drug resistant pulmonary tuberculosis among treated patients or on those on treatment without adequate response and (2) to evaluate HIV seropositivity among MDR-TB patients. METHODS: Pulmonary TB patients, who had at least six months of unsuccessful anti-tuberculous treatment were selected for the study. Their sputum specimens were examined for M. tuberculosis culture and drug sensitivity pattern and serological examinations for HIV infection were carried out. RESULTS: Sputum specimens of 618 patients' (61.8%) of a total of 1000 examined had shown culturable M. tuberculosis. Four hundred ninty-five patients (49.5%) were found to expectorate tubercle bacilli resistant to one or more anti TB drugs. MDR-TB was detected in 339 patients (33.9%). HIV seropositivity among MDR-TB was 4.42%. Significantly, 245 patients (24.5%) had tubercle bacilli resistant to one or more reserve drugs too (ethionamide, kanamycin and/or ofloxacin). CONCLUSIONS: Prevalence of MDR-TB was high in the study population. It is essentially an acquired condition. Its association with HIV disease was at present on the lower side, an observation contrary to published western literature. Higher rates of resistance for reserve drugs (ethionamide, kanamycin and/or ofloxacin) in patients who never had these drugs in their earlier treatment schedules suggest the possibility of emerging spontaneous drug resistant mutants.

6912. Deodhar L ,Goghate A .HIV status of culture positive tuberculosis cases.Indian J med Microbiol 1998,16(2).84-5. (ISA 013340, Vol 38 No13 ,1 July 2002)

6913. Dhar S; Dhar S.Histopathological features of granulomatous skin diseases : an analysis of 22 skin biopsies Indian Journal of Dermatology . 2002; 47(2): 88-90

6914. Efrati O, Barak A. Pleural effusions in the pediatric population. Pediatr Rev. 2002 Dec;23(12):417-26. No abstract.

6915. Furst DE, Cush J, Kaufmann S, Siegel J, Kurth R.Preliminary guidelines for diagnosing and treating tuberculosis in patients with rheumatoid arthritis in immunosuppressive trials or being treated with biological agents. Ann Rheum Dis. 2002 Nov;61 Suppl 2:ii62-3. No abstract.

6916. Gayathri Devi DR; Belwadi S; Beena PM..Diagnostic value of IgA in tuberculosis Indian Practitioner. 2002 Feb; 55(2): 97-9

6917. Goel A Seth A . Genitourinary Tuberculosis .Hosp Today 2002,7(2),75-8 (ISA 014380, Vol 38 No14 ,16 July 2002)

6918. Gupta S; Shende N; Banerjee S; Kumar S; Reddy MVR; Harinath BC. Analysis of seva TBES-31 antigen specific immunoglobulins IgM, IgA and IgG in sera of sputum and culture positive pulmonary tuberculosis Indian Journal of Clinical Biochemistry. 2002 Jan; 17(1): 5-8

6919. Hancox M.Cattle TB crisis--cause and cure, and further risk to human health. Respir Med. 2002 Oct;96(10):842-5. No abstract.

6920. Jasmer RM, Gotway MB, Creasman JM, Webb WR, Edinburgh KJ, Huang L.Clinical and radiographic predictors of the etiology of computed tomography-diagnosed intrathoracic lymphadenopathy in HIV-infected patients. J Acquir Immune Defic Syndr. 2002 Nov 1;31(3):291-8.

In HIV-infected patients with intrathoracic lymphadenopathy, it is not known whether clinical and radiographic findings are useful in predicting a specific diagnosis. We determined the etiology and predictors of the etiology of computed tomography (CT)-diagnosed intrathoracic lymphadenopathy in HIV-infected patients evaluated from June 1993 through April 1999. Multivariate analyses were performed to determine clinical and radiographic predictors of the three most common diagnoses. Of 318 patients, 110 (35%) had lymphadenopathy on chest CT. Among these 110 patients, tuberculosis/nontuberculous mycobacterial disease ( = 31), bacterial pneumonia ( = 26), and lymphoma ( = 21) were the most common diagnoses. Multivariate analysis identified cough and necrosis of lymph nodes on chest CT as independent predictors of tuberculosis/nontuberculous mycobacterial disease. African-American race, symptoms for 1 to 7 days, dyspnea, and presence of airways disease on chest CT were independent predictors of bacterial pneumonia; symptoms for >7 days, absence of cough, and absence of pulmonary nodules on CT independently predicted lymphoma. Intrathoracic lymphadenopathy is a frequent chest CT finding in HIV-infected patients. Opportunistic infections and lymphoma are the most common causes, and specific clinical and radiographic features can suggest these particular diagnoses.

6921. Jasmer RM, Nahid P, Hopewell PC. Clinical practice. Latent tuberculosis infection. N Engl J Med. 2002 Dec 5;347(23):1860-6. No abstract.

6922. Jasmer RM, Saukkonen JJ, Blumberg HM, Daley CL, Bernardo J, Vittinghoff E, King MD, Kawamura LM, Hopewell PC.Short-course rifampin and pyrazinamide compared with isoniazid for latent tuberculosis infection: a multicenter clinical trial. Ann Intern Med. 2002 Oct 15;137(8):640-7.

BACKGROUND: Rifampin and pyrazinamide are recommended for treatment of latent tuberculosis infection in adults without HIV infection, but reports of severe hepatotoxicity have raised concerns about its safety. Clinical trials have not compared this treatment with isoniazid in adults without HIV infection. OBJECTIVE: To compare the safety and tolerance of a 2-month regimen of rifampin and pyrazinamide with that of a 6-month regimen of isoniazid for treatment of latent tuberculosis infection. DESIGN: Multicenter, prospective, open-label trial. SETTING: Three urban public health tuberculosis clinics in the United States. PATIENTS: 589 adults with latent tuberculosis infection who met U.S. criteria for treatment. INTERVENTION: Patients were assigned in alternate weeks to receive rifampin and pyrazinamide daily for 2 months (n = 307) or isoniazid daily for 6 months (n = 282). MEASUREMENTS: Primary end points were hepatotoxicity, other adverse events, and percentage of patients who completed treatment. RESULTS: Sixteen of 207 (7.7%) patients assigned to rifampin and pyrazinamide developed grade 3 or 4 hepatotoxicity compared with 2 of 204 (1%) patients assigned to isoniazid (odds ratio, 8.46 [95% CI, 1.9 to 76.5]; P = 0.001). The rifampin plus pyrazinamide regimen was more likely than the isoniazid regimen to be discontinued because of hepatotoxicity (odds ratio, 5.19; P = 0.033). The overall percentage of nonhepatotoxic adverse events was 20% in the rifampin-pyrazinamide group and 16% in the isoniazid group. The proportion of patients who completed the study treatment was 61% and 57%, respectively. CONCLUSIONS: A 2-month regimen of rifampin and pyrazinamide was associated with an increased risk for grade 3 or 4 hepatotoxicity compared with a 6-month regimen of isoniazid. Liver enzymes should be measured routinely during treatment to screen for liver injury and prevent progression to severe toxicity.

6923. Julian E, Matas L, Perez A, Alcaide J, Laneelle MA, Luquin M. Serodiagnosis of tuberculosis: comparison of immunoglobulin A (IgA) response to sulfolipid I with IgG and IgM responses to 2,3-diacyltrehalose, 2,3,6-triacyltrehalose, and cord factor antigens. J Clin Microbiol. 2002 Oct;40(10):3782-8.

Nonpeptidic antigens from the Mycobacterium tuberculosis cell wall are the focus of extensive studies to determine their potential role as protective antigens or serological markers of tuberculous disease. Regarding this latter role and using an enzyme-linked immunosorbent assay, we have made a comparative study of the immunoglobulin G (IgG), IgM, and IgA antibody responses to four trehalose-containing glycolipids purified from M. tuberculosis: diacyltrehaloses, triacyltrehaloses, cord factor, and sulfolipid I (SL-I). Sera from 92 tuberculosis patients (taken before starting antituberculosis treatment) and a wide group of control individuals (84 sera from healthy donors, including purified protein derivative-negative, -positive, healed, and vaccinated individuals, and 52 sera from nontuberculous pneumonia patients), all from Spain, were studied. The results indicated a significantly elevated IgG and IgA antibody response in tuberculosis patients, compared with controls, with all the antigens used. SL-I was the best antigen studied, showing test sensitivities and specificities for IgG of 81 and 77.6%, respectively, and of 66 and 87.5% for IgA. Using this antigen and combining IgA and IgG antibody detection, high test specificity was achieved (93.7%) with a sensitivity of 67.5%. Currently, it is widely accepted that it is not possible to achieve sensitivities above 80% in tuberculosis serodiagnosis when using one antigen alone. Thus, we conclude that SL-I, in combination with other antigenic molecules, could be a useful antigen for tuberculosis serodiagnosis.

6924. Kang EY, Choi JA, Seo BK, Oh YW, Lee CK, Shim JJ.Utility of polymerase chain reaction for detecting Mycobacterium tuberculosis in specimens from percutaneous transthoracic needle aspiration. Radiology. 2002 Oct;225(1):205-9.

PURPOSE: To determine the clinical utility of polymerase chain reaction (PCR) for detecting Mycobacterium tuberculosis in specimens from percutaneous transthoracic needle aspiration (PTNA). MATERIALS AND METHODS: PCR for M tuberculosis detection in specimens from PTNA was performed prospectively in addition to cytologic and histologic analyses in 45 patients. On computed tomographic (CT) scans, tuberculosis (TB) versus malignant neoplasm or other infection was diagnosed in 28 patients; possible malignancy was diagnosed in 11, but TB was considered clinically because of young patient age or presence of tuberculous lesions in other areas of the lungs. In six of these patients, TB was diagnosed, but bacteriologic results were negative. PTNA was performed with a 21-gauge needle by one chest radiologist by using CT (n = 25), ultrasonographic (n = 5), or fluoroscopic guidance (n = 15). Final diagnoses were malignant neoplasm (n = 19), hamartoma (n = 1), TB (n = 17), and pneumonia (including actinomycosis and aspergillosis) (n = 8). Sensitivity, specificity, and positive predictive values of PCR in PTNA specimens for diagnosis of TB were calculated. RESULTS: In 17 patients with TB, PCR results were positive in 11 and negative in six. PCR results were negative in all cases of malignant neoplasm, hamartoma, and pneumonia. Cytologic and histologic analysis of PTNA specimens resulted in a specific diagnosis of TB in two (12%) of 17 patients. By adding the PCR results, diagnosis of TB was established in 12 (71%) of 17 patients. Sensitivity, specificity, and positive and negative predictive values of PCR for diagnosis of TB in PTNA specimens were 65% (11 of 17), 100% (28 of 28), 100% (11 of 11), and 82% (28 of 34), respectively. CONCLUSION: PCR for detection of M tuberculosis in PTNA specimens is a useful adjuvant to cytologic and histologic analysis for diagnosis of TB.

6925. Khan K, Muennig P, Behta M, Zivin JG.Global drug-resistance patterns and the management of latent tuberculosis infection in immigrants to the United States. N Engl J Med. 2002 Dec 5;347(23):1850-9.

BACKGROUND: In the United States, an increasingly disproportionate burden of tuberculosis among the foreign-born population has led to calls for improvements in the detection and treatment of latent infection in new immigrants. Current treatment guidelines do not take into account global differences in drug-resistance patterns or their implications for the treatment of immigrants. The use of multinational surveillance systems to guide the management of latent infection according to region-specific drug-resistance profiles could improve the efficiency of efforts to reduce the burden of tuberculosis in immigrants to the United States. METHODS: We constructed a decision-analysis model by using a hypothetical cohort of all documented immigrants entering the United States from developing nations. Region-specific drug-resistance profiles were derived from data on 30,388 cases of infection. The model examined the effectiveness and cost effectiveness of four strategies: no intervention or tuberculin skin testing followed by treatment with isoniazid, treatment with rifampin, or treatment with rifampin plus pyrazinamide for those with a positive test result. RESULTS: A strategy of detecting and treating latent tuberculosis infection was cost-saving among immigrants from Mexico, Haiti, sub-Saharan Africa, South Asia, and developing nations in East Asia and the Pacific. This strategy was highly cost effective among immigrants from other developing nations. Rifampin plus pyrazinamide was the preferred strategy for treating latent infection in immigrants from Vietnam, Haiti, and the Philippines. CONCLUSIONS: For new immigrants to the United States from developing nations, a strategy of detecting and treating latent tuberculosis infection would lead to substantial health and economic benefits. Because of the high prevalence of resistance to isoniazid, treatment with a rifampin-containing regimen should be strongly considered for immigrants from Vietnam, Haiti, and the Philippines. Copyright 2002 Massachusetts Medical Society

6926. Khan SA, Zahid M, Asif N, Hasan AS.Tuberculosis of the sternoclavicular joint. Indian J Chest Dis Allied Sci. 2002 Oct-Dec;44(4):271-3.

A rare case of tuberculosis of the stemoclavicular joint in a 13-year-old girl is presented. The occurrence of tubercular infection in the sternoclavicular joint is extremely rare.

6927. Khare K C , Datawani N .HIV infection amongest pulmonary tuberculosis.Hosp Today 2002,7(2),91-3. (ISA 014390, Vol 38 No14 ,16 July 2002)

6928. Khatri GR, Frieden TR. Controlling tuberculosis in India. N Engl J Med. 2002 Oct 31;347(18):1420-5.

BACKGROUND: Tuberculosis kills nearly 500,000 people in India each year. Until recently, less than half of patients with tuberculosis received an accurate diagnosis, and less than half of those received effective treatment. METHODS: We analyzed the effects of new policies introduced in 1993 that have resulted in increased resources, improved laboratory-based diagnosis, direct observation of treatment, and the use of standardized antituberculosis regimens and reporting methods. RESULTS: By September 2001, more than 200,000 health workers had been trained, and 436 million people (more than 40 percent of the entire population) had access to services. About 3.4 million patients had been evaluated for tuberculosis, and nearly 800,000 had received treatment, with a success rate greater than 80 percent. More than half of all those treated in the past 8 years were treated in the past 12 months. CONCLUSIONS: India's tuberculosis-control program has been successful in improving access to care, the quality of diagnosis, and the likelihood of successful treatment. We estimate that the improved program has prevented 200,000 deaths, with indirect savings of more than $400 million--more than eight times the cost of implementation. It will be a substantial challenge to sustain and expand the program, given the country's level of economic development, limited primary health care system, and large and mostly unregulated private health care system, as well as the dual threats of the human immunodeficiency virus and multidrug-resistant tuberculosis. Copyright 2002 Massachusetts Medical Society

6929. Kitada S, Maekura R, Toyoshima N, Fujiwara N, Yano I, Ogura T, Ito M, Kobayashi K. Serodiagnosis of pulmonary disease due to Mycobacterium avium complex with an enzyme immunoassay that uses a mixture of glycopeptidolipid antigens. Clin Infect Dis. 2002 Dec 1;35(11):1328-35.

It is difficult to distinguish pulmonary disease due to Mycobacterium avium complex (MAC) from that due to other mycobacteria, such as Mycobacterium tuberculosis and Mycobacterium kansasii. We developed an enzyme immunoassay (EIA) for diagnosis of MAC pulmonary diseases that uses glycopeptidolipid (GPL) antigens specific for MAC, and we used it for diagnosis in immunocompetent patients. The mean optical densities (+/- standard deviation) of serum immunoglobulin G antibodies to GPLs in patients with MAC disease, MAC colonization, M. kansasii disease, and tuberculosis and in healthy subjects were 0.778+/-0.784, 0.042+/-0.035, 0.059+/-0.035, 0.071+/-0.035, and 0.030+/-0.027, respectively. A significant increase in the level of anti-GPL antibodies was detected in patients with MAC disease. The level of anti-GPL antibodies reflected disease activity, because the level was decreased in culture-negative patients who had conversion of culture results. When a cutoff level of seropositivity (0.119) was defined, the sensitivity of EIA for diagnosis of MAC disease was 92.3%, and the specificity was 96.7%. Measurement of serum anti-GPL antibodies is useful for both the diagnosis of and assessment of activity in MAC disease.

6930. Kulkarni AG; Prabhu VV. Tuberculosis presenting as carpal tunnel syndrome: a case report with review of literature Bombay Hospital Journal. 2002 Jan; 44(1): 102-3

ABSTRACT: We present a case of tuberculosis presenting as carpal tunnel syndrome. Though it is not unusual, the aim is to draw attention that tuberculosis should be considered in the differential diagnosis on finding nonspecific tenosynovitis during carpal tunnel release and should be excluded by performing a biopsy.

6931. Kurth R, Haas WH.Epidemiology, diagnostic possibilities, and treatment of tuberculosis.Ann Rheum Dis. 2002 Nov;61 Suppl 2:ii59-61. No abstract.

6932. Larsen NM, Biddle CL, Sotir MJ, White N, Parrott P, Blumberg HM. Risk of tuberculin skin test conversion among health care workers: occupational versus community exposure and infection. Clin Infect Dis. 2002 Oct 1;35(7):796-801.

A prospective observational cohort study to assess rates of and risk factors for tuberculin skin test (TST) conversion among health care workers (HCWs) was conducted at an urban hospital located in a high tuberculosis-incidence area in 1994-1998. All hospital employees undergoing required testing every 6 months were included. A total of 69 (1.2%) of 5773 susceptible employees had a documented TST conversion (overall rate, 0.38 per 100 person-years worked). No significant difference existed in conversion rates among employees with frequent, limited, or no patient contact. HCWs with a TST conversion lived in zip codes with higher tuberculosis case rates (P< or =.05). In multivariate analysis, TST conversion was associated with history of bacille Calmette-Guerin vaccination (relative risk [RR], 11.63), annual salary <$20,000 (RR, 3.67), and increasing age. In the setting of an effective tuberculosis infection-control program, TST conversion rates were low, and risk of conversion among HCWs was associated most strongly with nonoccupational factors.

6933. Lee JH, Lee CW, Lee SG, Yang HS, Hong MK, Kim JJ, Park SW, Chi HS, Park SJ. Comparison of polymerase chain reaction with adenosine deaminase activity in pericardial fluid for the diagnosis of tuberculous pericarditis. Am J Med. 2002 Oct 15;113(6):519-21. No abstract.

6934. McCaughan F, Holmes A, Lynn WA, Friedland JS.Mycobacterium tuberculosis infection complicated by Eales disease with peripheral neuropathy. Clin Infect Dis. 2002 Oct 15;35(8):e89-91.

Eales disease, which is reported mainly in patients from the Indian subcontinent, is characterized by ophthalmic abnormalities that are sometimes followed by neurologic sequelae, and it is associated with previous Mycobacterium tuberculosis infection. We describe the first patient, to our knowledge, to receive a diagnosis of active tuberculosis and concurrent, severe neurological Eales disease, including peripheral neuropathy. The patient recovered completely after receiving steroid therapy.

6935. Michael JS; Lalitha MK; Cherian T; Thomas K; Mathai D; Abraham OC; Brahmadattan KN Evaluation of polymerase chain reaction for rapid diagnosis of tuberculosis meningitis Indian Journal of Tuberculosis. 2002 Jul; 49(3): 133-7

ABSTRACT: Background: Rapid and specific diagnosis of tuberculous meningitis is of utmost importance. Aim: To evaluate polymerase chain reaction (PCR) using primers directed against the IS6110 gene of M.tuberculosis in the diagnosis of tuberculous meningitis (TBM). Method: PCR was performed on CSF samples from 34 patients suspected to have tuberculous meningitis (TBM) and 68 likely to be having meningitis due to other causes(controls). The first group was further divided into definite TBM and probable TBM using culture as the gold standard. Results: In 17 culture positive patients (definite TBM), PCR was positive in 13 while in 17 patients who were diagnosed clinically, PCR picked up 11 cases all of whom were culture negative. There were 2 PCR positives among the control group: one patient was HIV positive having cryptococcal meningitis, with disseminated tuberculosis in whom true positivity could not be ruled out. For final diagnosis of tuberculous meningitis, the overall sensitivity of microscopy, culture and PCR were 3 percent, 50 percent and 71 percent and specificity 100 percent, 100 percent and 97 percent respectively. Conclusion: PCR is valuable in the rapid diagnosis of tuberculous meningitis.

6936. Mohapatra PR, Janmeja AK, Saini V, Das SK, Deb A.Second-line treatment for chronic tuberculosis.Lancet. 2002 Nov 2;360(9343):1430. No abstract.

6937. Mwinga A, Nunn A, Ngwira B, Chintu C, Warndorff D, Fine P, Darbyshire J, Zumla A. Mycobacterium vaccae (SRL172) immunotherapy as an adjunct to standard antituberculosis treatment in HIV-infected adults with pulmonary tuberculosis: a randomised placebo-controlled trial. Lancet. 2002 Oct 5;360(9339):1050-5.

BACKGROUND: Mortality rates of HIV-infected patients treated for tuberculosis remain high. This study aimed to assess the effect on mortality of immunotherapy with single-dose SRL172 added to standard antituberculosis chemotherapy in such patients. METHODS: The double-blind trial enrolled 1229 patients aged 18-60 years, who had never received antiretroviral treatment and who presented with newly diagnosed, sputum-smear-positive pulmonary tuberculosis to referral centres in Lusaka, Zambia, and Karonga, Malawi. Both HIV-positive and HIV-negative patients were enrolled, to avoid stigmatisation. Participants were randomly assigned a single injection of SRL172 or matching placebo within 2 weeks of starting 8 months of antituberculosis chemotherapy and followed up for at least 12 months. The primary endpoint was time to death in the HIV-infected population. Analyses were based on 760 HIV-positive patients after exclusion of 84 patients with errors in storage of the injection, no bacteriological confirmation, or no HIV result. FINDINGS: Of 760 HIV-infected patients, 374 received SRL172 and 386 received placebo. SRL172 did not cause any serious adverse events. The follow-up rate was 88% at 12 months in both groups. Of the HIV-positive patients, 109 (19.5 per 100 person-years) of 372 assigned SRL172 and 107 (19.3 per 100 person-years) of 386 assigned placebo died. In the Cox's regression analysis, stratified by centre, the hazard ratio of deaths (SRL172/placebo) was 1.03 (95% CI 0.79-1.35). There was no evidence of benefit to the group assigned SRL172. INTERPRETATION: Immunotherapy with single-dose SRL172 as an adjunct to standard antituberculosis treatment in HIV-positive adults with pulmonary tuberculosis had no significant effect on survival or bacteriological outcome, though the treatment was safe and well tolerated.

6938. Raut WK; Sawaitu VK; Bobhate SK; Fule RP; Salodkar AD.Acute miliary tuberculosis of skin-a case report and review of literature Indian Journal of Dermatology . 2002 Jan-Mar; 47(1): 57-9

ABSTRACT: Acute miliary tuberculosis of skin is a rare manifestation of tuberculosis. A 7 month old female presented with multiple subcutaneous nodules all over the body. Fine needle aspiration cytology and biopsy from the nodule showed tuberculous lesion with demonstration of acid-fast bacilli (AFB). Atypical presentation of cutaneous tuberculosis in HIV era and its diagnosis by AFB stain is stressed.

6939. Rege SA; Umman P; Nunes Q; Joshi A; Rohandia O.Rectal tuberculosis simulating malignancy: a case report and review Bombay Hospital Journal. 2002 Apr; 44(2): 280-1 .

ABSTRACT: Tuberculosis is known to involve any segment of gastrointestinal tract, however involvement distal to ileocaecal junction is rare. We report a case of rectal tuberculosis, which simulated malignancy, clinically and radiologically. Histopathology confirmed the diagnosis. Though treated with antitubercular treatments, patient had to be subjected to definitive surgical intervention.

6940. Senthil Kumar KS, Uma Devi KR, Alamelu R.Isolation and evaluation of diagnostic value of two major secreted proteins of Mycobacterium tuberculosis. Indian J Chest Dis Allied Sci. 2002 Oct-Dec;44(4):225-32.

Two secreted antigens of Mycobacterium tuberculosis, namely the antigen 85 complex (30/31) and 38kDa antigens, were purified from the whole culture filtrate by using two dimensional preparative electrophoresis and anion exchange chromatography, respectively. Individual components of the antigen 85 complex namely, antigen 85A, 85B and 85C, were separated using hydrophobic interaction chromatography. The humoral antibody activity to these antigens in sputum positive cases of active pulmonary tuberculosis and normal healthy volunteers was determined by enzyme linked immunosorbent assay (ELISA) and immunoblot. Recombinant 38kDa and antigen 6 were used as reference antigens for the assay. None of the healthy volunteers reacted with the 38kDa antigen, while 52% of the TB sera reacted with it. Of the three components of the antigen 85 complex, 85B gave the highest positivity of 40 per cent. The results of combination of 38kDa with antigen 6 offered better results with 76% positivity.

6941. Shenai S; Rodrigues C.Evaluation a new phage amplification technology for rapid diagnosis of tuberculosis Indian Journal of Medical Microbiology. 2002 Oct; 20(4): 194-90

ABSTRACT: Purpose: Rapid diagnosis of tuberculosis is essential to initiate timely and appropriate treatment to curb the spread of this potentially life threatening disease. The purpose of this study was to evaluate a phage amplification technology viz., FAST Plaque TB, TM for the diagnosis of tuberculosis. Methods: We evaluated the clinical utility of this new assay by analyzing 50 respiratory and 40 non-respiratory specimens, using FAST Plaque TBTM kit (Biotec Laboratories, UK) and the performance was compared with TB Bactec 460 semi-automated liquid culture system and conventional LJ culture method. Results: In case of respiratory specimens phage assay gave good specificity (100 percent) compared with TB Bactec whereas with respect to LJ method the sensitivity and specificity were 93.1 percent and 88.2 percent respectively. In case of non-respiratory specimens comparison of results obtained by phage assay showed sensitivity of 90.9 percent and specificity of 88.8 percent with respect to TB Bactec and 87.5 percent and 93.8 percent with respect to LJ method. Conclusions: We believe that this new low cost assay may have widespread applicability, especially in developing countries, due to its] manual format and rapid reporting of results.

6942. Venugopal P; Karunakaran R. Alappuzha Imaging diagnosis: a case report Indian Practitioner. 2002 Jan; 55(1): 46-8

ABSTRACT: A 43 year old lady presented with atypical radiological features of Bronchiolo alveolar carcinoma. Because of the unusual radiological picture and failure to recognize the symptom of bronchorrhoea, a proper diagnosis was delayed and the patient was misdiagnosed and treated as pulmonary tuberculosis.

Pathogenesis:

6943. Chan ED, Iseman MD. Current medical treatment for tuberculosis.BMJ 2002 Nov 30;325(7375):1282-6. No abstract.

6944. Cockle PJ, Gordon SV, Lalvani A, Buddle BM, Hewinson RG, Vordermeier HM. Identification of novel Mycobacterium tuberculosis antigens with potential as diagnostic reagents or subunit vaccine candidates by comparative genomics. Infect Immun 2002 Dec;70(12):6996-7003

An independent review for the British government has concluded that the development of a cattle vaccine against Mycobacterium bovis holds the best long-term prospects for tuberculosis control in British herds. The development of complementary diagnostic tests to differentiate between vaccinated and infected animals is necessary to allow the continuation of test-and-slaughter-based control policies alongside vaccination. Vaccination with M. bovis bacillus Calmette-Guerin (BCG), the only available vaccine, results in tuberculin purified protein derivative sensitivity and has shown varying vaccine efficacies in cattle. Thus, identification of more-specific reagents to distinguish between vaccination and infection, as well as the identification of subunit vaccine candidates for improved tuberculosis vaccines, is a research priority. In the present study, we applied comparative genomics to identify M. bovis-Mycobacterium tuberculosis antigens whose genes had been deleted in BCG Pasteur. In total, 13 open reading frames (ORFs) from the RD1, RD2, and RD14 regions of the M. tuberculosis genome were selected. Pools of overlapping peptides spanning these ORFs were tested in M. bovis-infected (n = 22), BCG-vaccinated (n = 6), and unvaccinated (n = 10) control cattle. All were recognized in infected cattle, with responder frequencies varying between 16 and 86%. In particular, eight highly immunogenic antigens were identified whose potentials as diagnostic reagents or as subunit vaccines warrant further study (Rv1983, Rv1986, Rv3872, Rv3873, Rv3878, Rv3879c, Rv1979c, and Rv1769).

6945. Gordon S.Positive about HIV--an immunological education project in South Africa. Nat Immunol 2002 Dec;3(12):1115-7. No abstract.

6946. Hughes-Davies TH. Changes in the transmission of tuberculosis in New York. N Engl J Med 2002 Oct 31;347(18):1453-5. No abstract.

6947. Larsen MH, Vilcheze C, Kremer L, Besra GS, Parsons L, Salfinger M, Heifets L, Hazbon MH, Alland D, Sacchettini JC, Jacobs WR Jr. Overexpression of inhA, but not kasA, confers resistance to isoniazid and ethionamide in Mycobacterium smegmatis, M. bovis BCG and M. tuberculosis. Mol Microbiol 2002 Oct;46(2):453-66

The inhA and kasA genes of Mycobacterium tuberculosis have each been proposed to ncode the primary target of the antibiotic isoniazid (INH). Previous studies nvestigating whether overexpressed inhA or kasA could confer resistance to INH ielded disparate results. In this work, multicopy plasmids expressing either nhA or kasA genes were transformed into M. smegmatis, M. bovis BCG and three ifferent M. tuberculosis strains. The resulting transformants, as well as reviously published M. tuberculosis strains with multicopy inhA or kasAB lasmids, were tested for their resistance to INH, ethionamide (ETH) or thiolactomycin (TLM). Mycobacteria containing inhA plasmids uniformly exhibited 20-fold or greater increased resistance to INH and 10-fold or greater increased resistance to ETH. In contrast, the kasA plasmid conferred no increased resistance to INH or ETH in any of the five strains, but it did confer resistance to thiolactomycin, a known KasA inhibitor. INH is known to increase the expression of kasA in INH-susceptible M. tuberculosis strains. Using molecular beacons, quantified inhA and kasA mRNA levels showed that increased inhA mRNA levels corre--lated with INH resistance, whereas kasA mRNA levels did not. In summary, analysis of strains harbouring inhA or kasA plasmids yielded the same conclusion: overexpressed inhA, but not kasA, confers INH and ETH resistance to M. smegmatis, M. bovis BCG and M. tuberculosis. Therefore, InhA is the primary target of action of INH and ETH in all three species.

6948. Orellana C. Immune system stimulator shows promise against uberculosis. Lancet Infect Dis 2002 Dec;2(12):711. No abstract.

6949. Quintana FJ, Carmi P, Cohen IR. DNA vaccination with heat shock protein 60 inhibits cyclophosphamide-accelerated diabetes. J Immunol 2002 Nov 15;169(10):6030-5

Nonobese diabetic (NOD) mice spontaneously develop diabetes as a consequence of an autoimmune process that can be inhibited by immunotherapy with the 60-kDa heat shock protein (hsp60), with its mycobacterial counterpart 65-kDa (hsp65), or with other Ags such as insulin and glutamic acid decarboxylase (GAD). Microbial infection and innate signaling via LPS or CpG motifs can also inhibit the spontaneous diabetogenic process. In addition to the spontaneous disease, however, NOD mice can develop a more robust cyclophosphamide- ccelerated diabetes (CAD). In this work, we studied the effect on CAD of DNA vaccination with constructs encoding the Ags human hsp60 (phsp60) or mycobacterial hsp65 (phsp65). Vaccination with phsp60 protected NOD mice from CAD. In contrast, vaccination with phsp65, with an empty vector, or with a CpG-positive oligonucleotide was not effective, suggesting that the efficacy of the phsp60 construct might be based on regulatory hsp60 epitopes not shared with its mycobacterial counterpart, hsp65. Vaccination with phsp60 modulated the T cellresponses to hsp60 and also to the GAD and insulin autoantigens; T cell proliferative responses were significantly reduced, and the pattern of cytokine secretion to hsp60, GAD, and insulin showed an increase in IL-10 and IL-5 secretion and a decrease in IFN-gamma secretion, compatible with a shift from a Th1-like toward a Th2-like autoimmune response. Our results extend the role of specific hsp60 immunomodulation in the control of beta cell autoimmunity and demonstrate that immunoregulatory networks activated by specific phsp60 vaccination can spread to other Ags targeted during the progression of diabetes, like insulin and GAD.

Vaccines :

6950. Hoft DF, Worku S, Kampmann B, Whalen CC, Ellner JJ, Hirsch CS, Brown RB, Larkin R, Li Q, Yun H, Silver RF. Investigation of the relationships between immune-mediated inhibition of mycobacterial growth and other potential surrogate arkers of protective Mycobacterium tuberculosis immunity. J Infect Dis. 2002 Nov 15;186(10):1448-57.

Therapy :

6951. Biet F, Kremer L, Wolowczuk I, Delacre M, Locht C.Mycobacterium bovis BCG producing interleukin-18 increases antigen-specific gamma interferon production in mice. Infect Immun. 2002 Dec;70(12):6549-57.

Interleukin-18 (IL-18) and IL-12 play a critical role in the expression of cell-mediated immunity involved in host defense against intracellular pathogens. Both cytokines are produced by macrophages and act in synergy to induce gamma interferon (IFN-gamma) production by T, B, and natural killer cells. In the present study, we analyzed both cellular and humoral responses upon infection with IL-18-secreting BCG of BALB/c and C3H/HeJ mice, two strains known to differ in their ability to support the growth of BCG. The cDNA encoding mature IL-18 was fused in frame with the alpha-antigen signal peptide-coding sequence, cloned downstream of the mycobacterial hsp60 promoter and expressed in BCG. IL-18 produced by the recombinant BCG strain was functional, as judged by NF-kappaB-mediated luciferase induction in a tissue culture assay. When susceptible mice were infected with IL-18-producing BCG, their splenocytes were found to produce higher amounts of Th1 cytokines after stimulation with mycobacterial antigens than the splenocytes of mice infected with the nonrecombinant BCG. This was most prominent for IFN-gamma, although the mycobacterial antigen-specific secretion of granulocyte-macrophage colony-stimulating factor and IL-10 was also augmented after infection with the recombinant BCG compared to infection with nonrecombinant BCG. In contrast, the immunoglobulin G levels in serum against mycobacterial antigens were lower when the mice were infected with IL-18-producing BCG compared to infection with nonrecombinant BCG. The IL-18 effect was delayed in BALB/c compared to C3H/HeJ mice. These results indicate that the production of IL-18 by recombinant BCG may enhance the immunomodulatory properties of BCG further toward a Th1 profile. This may be particularly useful for immunotherapeutic or prophylactic interventions in which a Th1 response is most desirable.

6952. Bose M. Genomics and tuberculosis. Indian J Chest Dis Allied Sci. 2002 Oct-Dec;44(4):221-3. No abstract.

6953. Collins CD, Green AT, Newell JN. The relationship between disease control strategies and health system development: the case of TB. Health Policy. 2002 Nov;62(2):141-60.

This paper focuses on the lack of dialogue and policy consonance between those taking the lead in health systems change and those developing specific disease control strategies. In the first part, the origins and characteristics of this situation are explained using, as an example, TB control. Attention is then paid to the development of disease control friendly health systems. Four aspects of policy development are analysed paying particular attention to TB control: analysis of policy context, mechanisms for collaboration between policy actors; agreement on decision-making processes; development of common aims and objectives. Although the focus is on TB control, the principles illustrated carry some relevance for other disease control programmes.

6954. Kang EY, Choi JA, Seo BK, Oh YW, Lee CK, Shim JJ. Utility of polymerase chain reaction for detecting Mycobacterium tuberculosis in specimens from percutaneous transthoracic needle aspiration. Radiology. 2002 Oct;225(1):205-9.

6955. Narita M, Kellman M, Franchini DL, McMillan ME, Hollender ES, Ashkin D. Short-course rifamycin and pyrazinamide treatment for latent tuberculosis infection in patients with HIV infection: the 2-year experience of a comprehensive community-based program in Broward County, Florida.Chest. 2002 Oct;122(4):1292-8.

OBJECTIVES: To determine the completion rate and tolerability of short-course rifamycin and pyrazinamide treatment of latent tuberculosis infection (LTBI) in HIV-infected patients through a comprehensive community-based program. DESIGN: Prospective cohort, with comparison to a historical control group. PATIENTS: Of 3,118 patients with HIV infection screened for LTBI between February 1999 and March 2001, 135 patients were placed on rifamycin/pyrazinamide for 2 months under directly observed therapy and were compared to a historical group comprised of 93 HIV-infected patients who were placed on self-administered treatment of isoniazid for 12 months between 1996 and 1998. RESULTS: Of 135 patients receiving rifamycin/pyrazinamide, 124 patients (92%) completed treatment; 5 patients had to discontinue treatment due to side effects (allergic skin reactions [n = 4], hepatitis [n = 1]). The completion rate of the historical group who received isoniazid therapy was 61% (57 of 93 patients; p < 0.001); none of those who received isoniazid experienced significant side effects. CONCLUSION: In our experience, a comprehensive, community-based program of rifamycin/pyrazinamide for LTBI achieved significantly higher adherence than that of traditional isoniazid therapy, and thus may provide improved tuberculosis prevention in a community with high prevalence of HIV-infected patients.

6956. Shukla SJ, Warren DK, Woeltje KF, Gruber CA, Fraser VJ.Factors associated with the treatment of latent tuberculosis infection among health-care workers at a midwestern teaching hospital. Chest. 2002 Nov;122(5):1609-14.

STUDY OBJECTIVE: To assess factors associated with initiating therapy and compliance with treatment for latent tuberculosis infection among health-care workers with positive tuberculin skin test results. DESIGN: Prospective cohort study. SETTING: An urban midwestern teaching hospital in St. Louis, MO. Study population: Health-care workers with positive tuberculin skin test results. MEASUREMENTS: (1) Rates of initiating therapy for latent tuberculosis infection among all health-care workers with positive tuberculin skin test results, and (2) compliance rates with therapy for latent tuberculosis infection among health-care workers with recent tuberculin skin test conversion. RESULTS: A total of 440 tuberculin skin test-positive health-care workers were evaluated from January 1, 1994, to May 1, 2000. Of those evaluated, 1 health-care worker had presumed active tuberculosis, 1 had no record of being evaluated, 1 had missing records, and 33 were not recommended isoniazid therapy, leaving 404 workers for analysis. Overall, 396 of 404 health-care workers (98%) with positive tuberculin skin test results initiated isoniazid therapy. In univariate analysis, bacille Calmette-Guerin (BCG) vaccination (p = 0.02) and foreign birth (p = 0.03) were significantly associated with not initiating isoniazid therapy. Compliance data were available for 388 of 404 health-care workers (96%). Of these, 318 of 388 health-care workers (82%) were compliant with 6 months of therapy. BCG vaccination (odds ratio [OR], 3.5; 95% confidence interval [CI], 1.8 to 7.1) and symptoms while receiving therapy (OR, 4.5; 95% CI, 2.0 to 10.1) were significantly associated with noncompliance in multivariate analysis. Among new converters, Asian race (p = 0.006), foreign birth (p = 0.01), BCG vaccination (p = 0.006), and symptoms while receiving therapy (p < 0.001) were significantly associated with noncompliance in univariate analysis. CONCLUSION: This hospital had a high rate of initiating isoniazid therapy for tuberculosis infection among their health-care workers, and a high rate of compliance with therapy. These rates of initiation and completion of isoniazid therapy were much higher than those previously reported in the literature. This may be largely due to a focused program, which includes active follow-up of health-care workers with positive tuberculin skin test results, consisting of physician counseling and monthly phone consultations by nurses, along with free services and medications provided on-site.

July 2003

7484. Agyekum S, Church A, Sohail M, Krausz T, Van Noorden S, Polak J, Cohen J. Expression of lymphotoxin-beta (LT-beta) in chronic inflammatory conditions. J Pathol. 2003 Jan;199(1):115-21.

Functional studies in gene-knockout and transgenic mice systems have shown that lymphotoxin-alpha and lymphotoxin-beta (LT-alpha and LT-beta) are of fundamental importance in peripheral lymphoid organ development, but it remains unclear what role these cytokines have to play in the adult immune response and in the pathogenesis of disease. In this study, a polyclonal anti-serum to human LT-beta was used to investigate the distribution of LT-beta by immunohistochemistry in normal and diseased tissues. In the gut, lymph nodes, spleen, and tonsil, there was some LT-beta present on a variety of lymphoid cell types. In contrast, strong staining for LT-beta was observed on plasma cells and a subpopulation of CD4+ T cells in tissues affected by chronic inflammatory disease or infection, for example in inflammatory bowel disease, and in lymph nodes obtained from patients with sarcoidosis and tuberculosis. In tuberculous and sarcoid lymph nodes, LT-beta expression also occurred on some but not all epithelioid histiocytes within granulomas and on multi-nucleated giant cells. These findings support a role for LT-beta in human disease and suggest that it might represent a therapeutic target in a variety of common infective or inflammatory disorders. Copyright 2002 John Wiley & Sons, Ltd.

7485. Ahmed Z, Sachan A S, Gupta R K. comparative evaluation of Elisa (A-60 test) with FNAC and mantoux test in the diagnosis of tuberculosis lymphadenitis. Indian med Gaz 2002, 136(2), 64-7. (19639)Vol 38 No 19 1 Oct 2002.

7486. Akahoshi M, Nakashima H, Miyake K, Inoue Y, Shimizu S, Tanaka Y, Okada K, Otsuka T, Harada M. Influence of interleukin-12 receptor beta1 polymorphisms on tuberculosis. Hum Genet. 2003 Mar;112(3):237-43.

Host genetic factors may be important determinants of susceptibility to tuberculosis, and several candidate gene polymorphisms have been shown to date. A series of recent reports concerning rare human deficiencies in the type-1 cytokine pathway suggest that more subtle variants of relevant genes may also contribute to susceptibility to tuberculosis at the general population level. To investigate whether polymorphisms in the interleukin-12 receptor (IL-12R) gene predispose individuals to tuberculosis, we studied these genes by single-strand conformational polymorphism analysis and direct sequencing. Although no common polymorphisms could be identified in the IL-12R beta 2 gene ( IL-12RB2), we confirmed four single nucleotide polymorphisms (SNPs; 641A-->G, 684C-->T, 1094T-->C, and 1132G-->C) causing three missense variants (Q214R, M365T, G378R) and one synonymous substitution in the extracellular domain of the IL-12R beta 1 gene ( IL12RB1). All SNPs were in almost perfect linkage disequilibrium (D'=0.98), and two common haplotypes of IL12RB1(allele 1: Q214-M365-G378; allele 2: R214-T365-R378) were revealed. Polymerase chain reaction/restriction fragment length polymorphism and sequence analyses were used to type IL12RB1polymorphisms in 98 patients with tuberculosis and 197 healthy controls in Japanese populations. In our case-control association study of tuberculosis, the R214-T365-R378 allele (allele 2) was over-represented in patients with tuberculosis, and homozygosity for R214-T365-R378 (the 2/2 genotype) was significantly associated with tuberculosis (odds ratio: 2.45; 95% CI: 1.20-4.99; P=0.013). In healthy subjects, homozygotes for R214-T365-R378 had lower levels of IL-12-induced signaling, according to differences in cellular responses to IL-12 between two haplotypes. These data suggest that the R214-T365-R378 allele, i.e., variation in IL12RB1, contribute to tuberculosis susceptibility in the Japanese population. This genetic variation may predispose individuals to tuberculosis infection by diminishing receptor responsiveness to IL-12 and to IL-23, leading to partial dysfunction of interferon-gamma-mediated immunity.

7487. Aoe K, Hiraki A, Murakami T, Eda R, Maeda T, Sugi K, Takeyama H. Diagnostic significance of interferon-gamma in tuberculous pleural effusions. Chest. 2003 Mar;123(3):740-4.

STUDY OBJECTIVES: Tuberculosis (TB), the single most frequent infectious cause of death worldwide, also is a major cause of pleural effusion, which in TB usually has lymphocytic and exudative characteristics. Differential diagnosis between TB and nontuberculous pleural effusion can be sometimes difficult, representing a critically important clinical problem. METHODS: We studied 46 patients presenting with pleural effusion to the National Sanyo Hospital between April 2000 and January 2001 (34 men and 12 women; mean age, 64 years). Ten patients (22%) had tuberculous pleurisy, 19 patients (41%) had malignant pleuritis, and 17 patients (37%) had pleural effusion due to an etiology other than tuberculosis or cancer. Pleural fluid concentrations of four suggested markers were measured using commercially available kits. RESULTS: The pleural fluid levels (mean +/- SE) of adenosine deaminase (83.3 +/- 18.2 U/L vs 25.8 +/- 20.4 U/L, p < 0.0001), interferon-gamma (137 +/- 230 IU/mL vs 0.41 +/- 0.05 IU/mL, p < 0.0001), immunosuppressive acidic protein (741 +/- 213 micro g/mL vs 445 +/- 180 micro g/mL, p < 0.001) and soluble interleukin 2 receptor (7,618 +/- 3,662 U/mL vs 2,222 +/- 1,027 U/mL, p < 0.0001) were significantly higher for tuberculous pleuritis than for other causes of effusion. Receiver operating characteristic analysis demonstrated that pleural fluid content INF-gamma was the best indicator of tuberculous pleurisy among four relevant biological markers. CONCLUSIONS: INF-gamma in pleural fluid is the most sensitive and specific among four biological markers for tuberculous pleuritis. Thus, our results suggest that determination of INF-gamma at the onset of pleural effusion is informative for the diagnosis of tuberculous pleuritis. Further studies including larger numbers of patients are needed to verify this result.

7488. Arunkumar M J, Rajshekar V. Intrasellar tuberculoma presenting as pituitary apoplexy. Neurol India 2001, 49(4), 407-10. (19650)Vol 38 No 19 1 Oct 2002.

7489. Bardarov S Jr, Dou H, Eisenach K, Banaiee N, Ya S, Chan J, Jacobs WR Jr, Riska PF. Detection and drug-susceptibility testing of M. tuberculosis from sputum samples using luciferase reporter phage: comparison with the Mycobacteria Growth Indicator Tube (MGIT) system. Diagn Microbiol Infect Dis. 2003 Jan;45(1):53-61. Rapid diagnosis of drug-resistant M.tuberculosis (Mtb) is desirable worldwide. We (i) describe a new luciferase reporter phage (LRP), phAE142 for this purpose; (ii) compare it to the automated MGIT 960 for time-to-detection of Mtb in clinical specimens; and (iii) evaluate its use for species confirmation and antibiotic susceptibility testing(AST) of Mtb. Twenty sputum samples were inoculated for testing by LRP, or by MGIT 960. After "positives" were identified by either method, the LRP was used for confirmation of Mtb complex (TBC) and for AST. The LRP method proved comparably efficient to MGIT 960 at detecting Mtb. Using an antibiotic uniquely inhibiting TBC with LRP provided species assignment, concurrently with AST, in a median of 3 days, with a sensitivity of 97%. Overall agreement in susceptibility results was 96%. Reliable susceptibility results and identification of TBC can be completed in a median of 12 days (range 8 to 16d) with LRP applied to sputum samples.

7490. Barthel R, Tsytsykova AV, Barczak AK, Tsai EY, Dascher CC, Brenner MB, Goldfeld AE. Regulation of tumor necrosis factor alpha gene expression by mycobacteria involves the assembly of a unique enhanceosome dependent on the coactivator proteins CBP/p300. Mol Cell Biol. 2003 Jan;23(2):526-33.

Tumor necrosis factor alpha (TNF-alpha) plays an important role in host containment of infection by Mycobacterium tuberculosis, one of the leading causes of death by an infectious agent globally. Using the pathogenic M. tuberculosis strain H37Rv, we present evidence that upon stimulation of monocytic cells by M. tuberculosis a unique TNF-alpha enhanceosome is formed, and it is distinct from the TNF-alpha enhanceosome that forms in T cells stimulated by antigen engagement or virus infection. A distinct set of activators including ATF-2, c-jun, Ets, Sp1, Egr-1 and the coactivator proteins CBP/p300 are recruited to the TNF-alpha promoter after stimulation with M. tuberculosis. Furthermore, the formation of this enhanceosome is dependent on inducer-specific helical phasing relationships between transcription factor binding sites. We also show that the transcriptional activity of CBP/p300 is potentiated by mycobacterial stimulation of monocytes. The identification of TNF-alpha regulatory elements and coactivators involved in M. tuberculosis-stimulated gene expression thus provides potential selective molecular targets in the modulation of TNF-alpha gene expression in the setting of mycobacterial infection.

7491. Bhattacharya B, Karak K, Ghosal AG, Roy A, Das S, Dandapat P, Khetawat D, Mondal DK, Bhattacharya S, Chakrabarti S. Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species. Trop Med Int Health. 2003 Feb;8(2):150-7.

For early detection and species differentiation of mycobacteria, polymerase chain reaction (PCR) techniques are currently in wide use. However, individual techniques using amplification of different targets with appropriate primers still have some limitations, which have to be overcome. The ideal technique would use DNA sequences which should be present in all mycobacteria and absent in others and would be able to discriminate one species from the other, as non-tuberculous mycobacteria (NTM) are on rise in terms of frequency of detection. We developed a multiplex PCR based on amplification of 165, 365 and 541 bp target fragments of unrelated genes, hsp 65 coding for 65 kDa antigen, dnaJ gene of mycobacteria and insertion element IS 6110 of Mycobacterium tuberculosis, respectively. This multiplex PCR was tested over 5 years from 1996 to 2001 with 411 clinical specimens from suspected cases of tuberculosis and mycobacterioses and compared with standard laboratory techniques. The multiplex PCR was positive for 379 cases compared with 280 cases by standard techniques (P < 0.0001). It could distinguish between strains of the M. tuberculosis complex and NTM; the results are comparable with standard techniques. Thus the multiplex PCR can be useful in early detection, species differentiation and epidemiology.

7492. Borelli V, Vita F, Shankar S, Soranzo MR, Banfi E, Scialino G, Brochetta C, Zabucchi G. Human eosinophil peroxidase induces surface alteration, killing, and lysis of Mycobacterium tuberculosis. Infect Immun. 2003 Feb;71(2):605-13.

The antimycobacterial role of eosinophil peroxidase (EPO), one of the most abundant granule proteins in human eosinophils, was investigated. Our data indicate that purified EPO shows significant inhibitory activity towards Mycobacterium tuberculosis H37Rv. On a molar basis, this activity was similar to that exhibited by neutrophil myeloperoxidase (MPO) and was both dose and time dependent. In contrast to the activity of MPO, which requires H(2)O(2), EPO also exhibited anti-M. tuberculosis activity in the absence of exogenously added peroxide. Morphological evidence confirmed that the mechanism of action of EPO against mycobacteria differs from that of MPO. While MPO kills M. tuberculosis H37Rv exclusively in the presence of hydrogen peroxide, it does not induce morphological changes in the pathogen. In contrast, EPO-treated bacteria frequently had cell wall lesions and eventually underwent lysis, either in the presence or in the absence of H(2)O(2).

7493. Byrd RP Jr, Mehta JB, Roy TM. Delay in diagnosis among hospitalized patients with active tuberculosis--predictors and outcomes. Am J Respir Crit Care Med. 2003 Jan 15;167(2):278 No abstract available

7494. Cherian G, Habashy AG, Uthaman B, Cherian JM, Salama A, Anim JT. Detection and follow-up of mediastinal lymph node enlargement in tuberculous pericardial effusions using computed tomography. Am J Med. 2003 Mar;114(4):319-22. No abstract available

7495. Deswal B S, Bhatnagar D, Kumar D, Deshpande V R. Prevalence of HIV in tuberculosis cases. Indian J Commun Med 2002, 27(2), 80-4. (21850) Vol 38, No. 21, 1 Nov 2002.

7496. Drobniewski FA, Caws M, Gibson A, Young D. Modern laboratory diagnosis of tuberculosis. Lancet Infect Dis. 2003 Mar;3(3):141-7.

One-third of the global population is believed to be infected with bacteria of the Mycobacterium tuberculosis complex, the causative agent of tuberculosis. More than 8 million new cases of tuberculosis occur annually leading to 2 million deaths. Mortality is particularly high in those coinfected with HIV and where the bacteria are multiple-drug-resistant strains--ie, strains resistant to at least isoniazid and rifampicin. Early diagnosis of tuberculosis and drug resistance improves survival and by identifying infectious cases promotes contact tracing, implementation of institutional cross-infection procedures, and other public-health actions. This review addresses significant advances made in the diagnosis of infection, clinical disease, and drug resistance over the past decade. It proposes operational criteria for a modern diagnostic service in the UK (as a model of a low-incidence country) and explores some of the economic issues surrounding the use of these techniques.

7497. Dutta B. chatterjee G, Das T K. Primary tuberculosis of the glans penis. Indian J Derm 2001, 46(4), 245-7. (20700) Vol 38, No. 20, 16 oct 2002.

7498. Eidlitz-Markus T, Zeharia A, Baum G, Mimouni M, Amir J. Use of the urine color test to monitor compliance with isoniazid treatment of latent tuberculosis infection. Chest. 2003 Mar;123(3):736-9.

STUDY OBJECTIVE: To apply the Arkansas color method in order to evaluate drug compliance and factors that can predict treatment adherence in patients being treated for latent tuberculosis infection (LTBI) with a single daily dose of isoniazid (INH). DESIGN: Prospective study of adherence of 105 patients aged 1 to 75 years who were treated with a single daily dose of INH for LTBI. INTERVENTIONS: Patients or their parents were interviewed regarding parameters that may affect compliance. Urine samples were collected and tested for INH metabolites with the Arkansas color method. RESULTS: Nonadherence to treatment was found in 28.5% of patients. There was no statistically significant correlation among the following parameters: gender; age; diagnosis; mode of administration (self or parents); duration of treatment; dose of INH per weight; or interval since last intake of dose. Twenty-six patients were randomly checked for treatment adherence on two separate visits, and nonadherent patients were informed immediately and their condition was fully explained to them. Five of six patients who were nonadherent in the first visit and were examined twice became adherent in the second visit. Three of 20 patients who were adherent in the first visit became nonadherent. CONCLUSION: Almost one third of the patients who received LTBI treatment with INH were nonadherent to treatment. No factor was found to predict adherence. The Arkansas method can be used by the family physician and is a simple, immediate method to follow-up patients with LTBI who are treated with INH.

7499. Erdal Yilmal, Gurgoze K M, Ilhan N, Dogan Y, Aydinoglu H. Interleukin-8 levels in children with tuverculosis and aseptic meningitis. Indian J Padiat 2002, 69(3), 219-21. (207020 Vol 38, No. 20, 16 oct 2002.

7500. Ezung T, Devin N T, Tombi Singh N, Biren Singh T. Pulmonary tuberculosis and diabetes mellitus- a study. J Indian Med Ass 2002, 100(6), 376-9. Vol 38, No. 20, 16 oct 2002. (24875) Vol 38, No. 20, 16 oct 2002.

7501. Ferreira WA, Mayrink W, dos Mares-Guia ML, Tavares CA.Detection and characterization of leishmania antigens from an American cutaneous leishmaniasis vaccine for diagnosis of visceral leishmaniasis. Diagn Microbiol Infect Dis. 2003 Jan;45(1):35-43.

Antigens were isolated from vaccines against American Cutaneous Leishmaniasis (ACL) and their reactivity tested against nine different groups of human sera and two groups of dog sera. These antigens react specifically with human and dog visceral leishmaniasis sera when compared to sera from non-infected individuals. Sera from humans from endemic areas of ACL before, or one year after, vaccination, and ACL patients treated and cured by immunotherapy with polyvalent vaccine, did not display significant differences of reactivity to these antigens. In contrast, they displayed a significantly higher reactivity to the antigens when compared to sera from healthy humans from non-endemic areas. No sera reactivity was observed with patients carrying Chagas' disease or tuberculosis. These antigens are polysaccharides aggregates and present molecular masses ranging from 90 to over 200 KDa. These data suggest the use of these antigens for sero-diagnosis of human and canine visceral leishmaniasis.

7502. Fritscher-Ravens A, Schirrow L, Pothmann W, Knofel WT, Swain P, Soehendra N. Critical care transesophageal endosonography and guided fine-needle aspiration for diagnosis and management of posterior mediastinitis. Crit Care Med. 2003 Jan;31(1):126-32.

BACKGROUND: Acute mediastinitis is a serious complication; it occurs after esophageal perforation and thoracic surgery and is rarely due to infections. Clinical and computed tomographic scan signs may be nonspecific, especially in postoperative patients. DESIGN: We prospectively evaluated the value of transesophageal endosonography with guided fine-needle aspiration in the diagnosis and identification of etiologic agents in critically ill patients with suspected posterior mediastinitis. SETTING: University hospital. PATIENTS AND METHODS: Transesophageal endosonography/fine-needle aspiration was performed at the bedside in the intensive care unit with a Pentax 34UX echo-endoscope and a portable Hitachi console (EUB 525). Eighteen patients with clinically suspected mediastinitis were examined with intensive care team support. RESULTS: Computed tomography was performed before transesophageal endosonography in all 18 patients and was inconclusive in 9. Transesophageal endosonography detected mediastinal lesions in 16 (89%) of 18 patients and was more accurately diagnostic than computed tomography (p =.0082). Fifteen patients had undergone surgery (11 esophagectomy, 1 other esophageal surgery, 1 head/neck cancer surgery, 1 complication after dilatational tracheostomy, and 1 with intervention after polytrauma). Three patients were suspected to have nonpostoperative mediastinitis. In 16 patients, infectious organisms were detected (bacterial, n = 14; fungal, n = 1; tuberculosis, n = 1). Culture and sensitivity of transesophageal endosonography/fine-needle aspiration specimens led to appropriate drug therapy. In two patients, methicillin-resistant Staphylococcus aureus was detected, leading to isolation care. Twelve patients improved; six died. Of the two patients in whom transesophageal endosonography did not detect a mediastinitis, one was false negative on autopsy. There were no complications. CONCLUSION: Bedside transesophageal endosonography/fine-needle aspiration of posterior mediastinal lesions in critically ill patients was an effective and relatively noninvasive way to detect mediastinitis and provide material to identify the etiologic agent. It was particularly useful in postesophagectomy patients.

7503. Gupta R C, Dave L, Gupta M L, Vats M, Gupta N. Primary tuberculosis of middle ear. Antiseptic, Madurai 2002, 99(7), 254. (24891) Vol 38, No. 20, 16 oct 2002.

7504. Gurbuz AK, Yazgan Y, Ozel MA, Cavuslu S, Altunay H, Yildirim S. Does high albumin gradient ascites accompany tuberculous peritonitis? J Clin Gastroenterol. 2003 Jan;36(1):82-3.No abstract available

7505. Harries AD, Hargreaves NJ, Kwanjana JH, Salaniponi FM. The diagnosis of extrapulmonary tuberculosis in Malawi. Trop Doct. 2003 Jan;33(1):7-11.

There is little information on a country-wide basis in sub-Saharan Africa about how the diagnosis of extra-pulmonary tuberculosis (EPTB) is made. A country-wide cross-sectional study was carried out in 40 non-private hospitals in Malawi which register and treat (TB) patients in order to assess diagnostic practices in adults registered with EPTB. All patients aged 15 years and above in hospital on treatment for EPTB were reviewed usingTB registers, case note files and clinical assessment. There were 244 patients, 132 men and 112 women whose mean age was 36 years. In 138 (57%) patients, all appropriate procedures and investigations, commensurate with hospital resources, had been carried out. Of 171 EPTB patients with cough for 3 weeks or longer, 138 (81%) submitted sputum specimens for smear microscopy of acid-fast bacilli (AFB). A confirmed diagnosis ofTB was made in 15 (6%) patients based on finding AFB or caseating granulomas in specimens. In 157 (64%) patients, the diagnosis of EPTB was considered to be correct. In 46 (19%) patients the diagnosis was considered to be TB, although different from the type of EPTB with which the patient was registered. In 39 (16%) patients an alternative non-TB diagnosis was made and in two (1%) patients it was not possible to make a decision. Diagnostic practices need to be improved, and ways of doing this are discussed.

7506. Hartman EE, Delbanco T. A 52-year-old man with a positive PPD, 1 year later. JAMA. 2003 Jan 22-29;289(4):476. No abstract available

7507. Kontos F, Petinaki E, Gitti Z, Costopoulos C, Anagnostou S, Tselentis I,Maniatis AN. Combined use of the fully automated Bactec MGIT 960 system and a PCR-restriction fragment length polymorphism analysis for routine detection and identification of mycobacteria from clinical samples. J Microbiol Methods. 2003 Jan;52(1):137-40.

The combined use of Bactec MGIT 960 system and a PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis was assessed for the rapid detection and identification of mycobacteria from clinical samples. The diagnostic sensitivity and the time of detection of MGIT 960 system were significantly higher than of Lowenstein-Jensen medium. PCR-RFLP identification analysis results were in concordance with those obtained by the conventional biochemical tests.

7508. Koppaka VR, Harvey E, Mertz B, Johnson BA.Risk factors associated with tuberculin skin test positivity among university students and the use of such factors in the development of a targeted screening program. Clin Infect Dis. 2003 Mar 1;36(5):599-607.

The present study evaluated the accuracy of a risk assessment questionnaire (RAQ) for identifying candidates for tuberculin testing. A 33-question RAQ was administered to students before they underwent tuberculin screening at Virginia Commonwealth University (Richmond). Test operating characteristics for the complete and abbreviated RAQs compared to tuberculin skin test (TST) results were determined. Of 5382 students screened, 125 (2.3%) had a positive TST result; 113 (90.4%) of these 125 students had >or=1 affirmative response on the RAQ (i.e., a "positive RAQ"). The prevalence of TST positivity among students not born in the United States was 33.2-fold higher than that among students born in the United States. A 2-question RAQ had a sensitivity of 81.6%, a specificity of 91.0%, and positive and negative predictive values of 17.7% and 99.5%, respectively. Risk assessment can be an accurate means of identifying candidates for tuberculin screening.

7509. Kuruvilla A. Ocular tuberculosis. Lancet. 2003 Jan 18;361(9353):260-1; No abstract available.

7510. Lata Kumar, Singh M. Respiratory allergy. Indian J Padiat 2002, 69(3), 237-44. No abstract available.

7511. Lewis KN, Liao R, Guinn KM, Hickey MJ, Smith S, Behr MA, Sherman DR. Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette- uerin attenuation. J Infect Dis. 2003 Jan 1;187(1):117-23.

The tuberculosis (TB) vaccine bacille Calmette-Guerin (BCG) is a live attenuated organism, but the mutation responsible for its attenuation has never been defined. Recent genetic studies identified a single DNA region of difference, RD1, which is absent in all BCG strains and present in all Mycobacterium tuberculosis (MTB) strains. The 9 open-reading frames predicted within this 9.5-kb region are of unknown function, although they include the TB-specific immunodominant antigens ESAT-6 and CFP-10. In this study, RD1 was deleted from MTB strain H37Rv, and virulence of H37Rv:DeltaRD1 was assessed after infections of the human macrophage-like cell line THP-1, human peripheral blood monocyte-derived macrophages, and C57BL/6 mice. In each of these systems, the H37Rv:DeltaRD1 strain was strikingly less virulent than MTB and was very similar to BCG controls. Therefore, it was concluded that genes within or controlled by RD1 are essential for MTB virulence and that loss of RD1 was important in BCG attenuation.

7512. Magadevan S. Serodiagnosis of tuberculosis is children. J Indian Med Ass 2002, 100(2), 99-102. (20760) Vol 38, No. 20, 16 oct 2002.

7513. Mazurek GH, Villarino ME. Guidelines for using the QuantiFERON-TB test for diagnosing latent Mycobacterium tuberculosis infection. Centers for Disease Control and Prevention. MMWR Recomm Rep. 2003 Jan 31;52(RR-2):15-8.

Until 2001, the only test used to diagnose latent tuberculosis infection (LTBI) was the tuberculin skin test (TST). However, in 2001, a new test (QuantiFERON-TB or QFT; manufactured by Cellestis Limited, Carnegie, Victoria, Australia) that measures the release of interferon-gamma in whole blood in response to stimulation by purified protein derivative was approved by the Food and Drug Administration. This statement provides interim recommendations for using and interpreting QFT. As with TST, interpretation and indicated applications of QFT differ for persons according to their risk for LTBI and for developing tuberculosis (TB). This report provides guidance for public health officials, health-care providers, and laboratorians with responsibility for TB control activities in the United States in their efforts to incorporate QFT testing for detecting and treating LTBI. Regardless of the test used to identify LTBI, testing should be primarily targeted at diagnosing infected patients who will benefit from treatment.

7514. Menzies D, Fanning A, Yuan L, FitzGerald JM. Factors associated with tuberculin conversion in Canadian microbiology and pathology workers. Am J Respir Crit Care Med. 2003 Feb 15;167(4):599-602.

The risk of occupational tuberculosis (TB) infection and associated factors was estimated among all microbiology and pathology technicians and compared with a sample of nonclinical personnel in 17 Canadian acute care hospitals. Participants underwent tuberculin skin testing and completed questionnaires. Prior skin tests and vaccinations and all patients with TB hospitalized in the preceding 3 years were reviewed. Of the work areas where direction of air flow and air changes per hour were measured, only 51% were adequately ventilated. Among participating lab workers the average annual risk of tuberculin conversion was 1.0%. This was associated with lower hourly air exchange rates (16.7 versus 32.5 in workers with no conversion, p < 0.001) work in pathology (adjusted odds ratio [OR]: 5.4; [95% confidence interval: 1.3, 22], higher proportion of patients with missed diagnosis in the first 24 hours (per 20% increase-OR: 2.0; [1.3, 3.2], treatment delayed 1 week or more (per 20% increase-OR: 2.0; [3.2, 3.2]), and higher mortality (per 20% increase-OR: 2.5; [1.1, 5.6]). We conclude that laboratory workers, with no direct patient contact, have increased risk of tuberculin conversion in hospitals where a greater proportion of patients with TB die, or have delayed, or missed diagnosis, although this may be modified by workplace ventilation.

7515. Menzies D. Sputum induction: simpler, cheaper, and safer--but is it better? Am J Respir Crit Care Med. 2003 Mar 1;167(5):676-7. No abstract available.

7516. Mishra OP, Batra P, Ali Z, Anupurba S, Das BK. Cerebrospinal fluid lysozyme level for the diagnosis of tuberculous meningitis in children.J Trop Pediatr. 2003 Feb;49(1):13-6.

Lysozyme activity was assayed in the cerebrospinal fluid (CSF) of 32 tuberculous meningitis (TBM), 17 bacterial meningitis, 10 partially treated bacterial meningitis, 18 encephalitis and 18 control subjects. The mean CSF lysozyme activity was significantly raised (p < 0.001) in TBM patients compared with other study groups. A cut-off CSF lysozyme level of > or = 26 U/l had a sensitivity and specificity of 93.7 and 84.1 per cent, respectively for the diagnosis of TBM. Overall, it was found to be a better test than any other single test and thus can be used for rapid and early diagnosis of TBM inchildren.

7517. Montenegro SH, Gilman RH, Sheen P, Cama R, Caviedes L, Hopper T, Chambers R, Oberhelman RA. Improved detection of Mycobacterium tuberculosis in Peruvian children by use of a heminested IS6110 polymerase chain reaction assay. Clin Infect Dis. 2003 Jan 1;36(1):16-23.

A novel heminested IS6110 polymerase chain reaction (PCR) assay was evaluated as a tool for diagnosing tuberculosis in 222 children. In an analysis of 392 specimens (gastric aspirates, nasopharyngeal aspirates, and sputum samples), results of PCR were compared with those of 3 culture methods, acid-fast bacillus (AFB) staining, and clinical assessment by the Stegen-Toledo score. The sensitivity of PCR (67%) was comparable to that of the 3-culture method (71%) and was significantly higher than that of Lowenstein-Jensen culture (54%) or AFB stain (42%) for children with highly probable tuberculosis. PCR detection rates for culture-positive specimens were 100% for smear-positive samples and 76.7% for smear-negative samples. The specificity of PCR was 100% in control children. Compared with culture, PCR demonstrated a sensitivity of 90.4%, a positive predictive value of 89%, a specificity of 94%, and a negative predictive value of 95% (kappa=.85). With clinical assessment as the standard, PCR had asensitivity of 71%, a positive predictive value of 92%, a specificity of 95%, and a negative predictive value of 79% (kappa=.67). PCR is a rapid and sensitive method for the early diagnosis of pediatric tuberculosis.

7518. Mustafa AS, Shaban FA, Al-Attiyah R, Abal AT, El-Shamy AM, Andersen P, Oftung F. Human Th1 cell lines recognize the Mycobacterium tuberculosis ESAT-6 antigen and its peptides in association with frequently expressed HLA class II molecules. Scand J Immunol. 2003 Feb;57(2):125-34.

We have used a synthetic-peptide approach to map epitope regions of the Mycobacterium tuberculosis ESAT-6 antigen recognized by human T cells in relation to major histocompatibility complex (MHC) restriction. ESAT-6-specific CD4+ T-cell lines were established by stimulating peripheral blood mononuclear cells from 25 HLA-DR-typed tuberculosis patients with complete antigen in vitro. The established T-cell lines were then screened for proliferation and interferon-gamma (IFN-gamma) secretion in response to eight overlapping 20-mer peptides covering the ESAT-6 sequence. The response of the T-cell lines to ESAT-6 and peptides from a human leucocyte antigen (HLA)-heterogeneous group of donors suggested the presence of multiple epitopes and promiscuous recognition of the antigen. Analysis of antigen and peptide recognition in the presence of anti-HLA class I and class II antibodies suggested that the T-cell lines recognized ESAT-6 in association with HLA-DR and -DQ molecules. Furthermore, testing of selected T-cell lines with ESAT-6 and the peptides in the presence of autologous and allogeneic HLA-DR- and -DQ-typed antigen-presenting cells identified HLA-DR2, -DR52 and -DQ2 amongst the HLA molecules involved in the presentation of ESAT-6 and its peptides to human Th1 cells. In addition, the T-cell lines were cytotoxic for monocytes and macrophages pulsed with ESAT-6 and peptides. In conclusion, the recognition of ESAT-6 by IFN-gamma-secreting and cytotoxic CD4+ T cells in association with frequently expressed HLA class II molecules supports the application of this antigen to either specific diagnosis or subunit vaccine design.

7519. Naik M B, Chaturvedi R M. Occupational profile of defaulting tuberculosis patients. Indian J Occup Hlth 2001, 44(4), 175-9. (20781)Vol 38, No. 20, 16 oct 2002

7520. Park do Y, Kim JY, Choi KU, Lee JS, Lee CH, Sol MY, Suh KS. Comparison of polymerase chain reaction with histopathologic features for diagnosis of tuberculosis in formalin-fixed, paraffin-embedded histologic specimens. Arch Pathol Lab Med. 2003 Mar;127(3):326-30.

Objective.-To investigate the relationship between various histopathologic features and the results of the tuberculosis (TB)-polymerase chain reaction (PCR) method in routinely submitted histologic specimens for the histopathologic diagnosis of TB. Design.-We used 95 formalin-fixed, paraffin-embedded tissue blocks from 81 patients who were clinically suspected of having TB. We assessed the presence of histopathologic features including well-formed granuloma, poorly formed granuloma, caseous necrosis, and Langhans-type giant cells. We performed nested PCR for IS6110 and Ziehl-Neelsen staining for acid-fast bacilli (AFB). Results.-Of the 81 patients studied, 53 patients had chronic granulomatous inflammation, whereas 28 patients had only chronic inflammation without definite granulomatous inflammation. Of the 53 cases with chronic granulomatous inflammation, 17 (32%) were AFB positive and 36 (68%) were TB-PCR positive. Among cases with chronic granulomatous inflammation, the percentage that were positive and negative by TB-PCR differed significantly with the presence of various histopathologic features. All of the 13 cases with well-formed granuloma, caseous necrosis, and Langhans-type giant cells were TB-PCR positive; however, 10 (36%) of the 28 cases with chronic inflammation without granulomatous lesions were also TB-PCR positive. Conclusions.-TB-PCR is a rapid, sensitive method for the diagnosis of TB in routinely processed formalin-fixed, paraffin-embedded histologic specimens and is readily available in histopathology laboratories. We recommend use of TB-PCR when TB is suspected clinically, especially in cases of chronic inflammation without definite evidence of granulomatous inflammation.

7521. Perkins MD, Conde MB, Martins M, Kritski AL.Serologic diagnosis of tuberculosis using a simple commercial multiantigen assay. Chest. 2003 Jan;123(1):107-12.

SETTING: Seven primary health clinics and a pulmonary disease specialty clinic in Rio de Janeiro City, Brazil. OBJECTIVE: To evaluate a commercial immunochromatographic test kit (ICT Tuberculosis; AMRAD-ICT; Sidney, Australia) employing five recombinant Mycobacterium tuberculosis proteins for the detection of pulmonary tuberculosis (TB). DESIGN: Serology test results were compared with duplicate sputum microscopy and culture in 277 patients with symptomatic pulmonary disease (243 with pulmonary TB and 34 with nontuberculous disease). An additional 110 healthy control subjects were also tested. RESULTS: The serology test was simple and rapid to perform and detected 64.2% of smear-positive and 46.3% of smear-negative TB patients overall. HIV co-infection was present in 15.3% of TB patients, and serology was much less sensitive (overall 27.6%) in this small group, as was microscopy (13.8%). Specificity of the serology test was 100% in healthy control subjects and 85.2% in the small number of control patients with pulmonary disease, including those with prior TB. Combined with microscopy, serology detected 72.8% of TB patients. CONCLUSION: Depending on the population studied, multiantigen serologic tests for TB may be as sensitive as microscopy, but detect a different and overlapping subset of patients. The use of multiple antigens in this kit increased test sensitivity without significant loss of specificity. Bacille Calmette-Guerin vaccination and tuberculin sensitivity did not affect serology results. Estimating specificity in clinical use will require testing a much larger cohort of symptomatic patients with nontuberculous disease. The TB diagnostic performance of this group of antigens in HIV co-infected individuals was poor.

7522. Rajesh M, Sulochana KN, Sundaram AL, Krishnakumar S, Biswas J, Ramakrishnan S. Presence of a 88 kDa Eales protein in uveitis, tuberculosis, leprosy and rheumatoid arthritis. Med Sci Monit. 2003 Feb;9(2):CR95-9.

BACKGROUND: Eales disease (ED) is an idiopathic retinal vasculitis affecting young adult males. We have earlier reported the identification, purification and partial characterization of a novel 88 kDa protein found in the serum of patients with ED. The aim of the present study was to look for the 88 kDa protein in serum samples obtained from cases of retinal vasculitis mimicking ED and in other systemic inflammatory diseases. MATERIAL/METHODS: Serum samples from healthy volunteers and from patients with ED, uveitis, parsplanitis ocular sarcoidosis, toxoplasmosis, leprosy, diabetic retinopathy, viral hepatitis, and rheumatoid arthritis were analyzed for the presence of the 88 kDa protein by polyacralymide gel electrophoresis (PAGE). The immunological identity of the 88 kDa protein found in ED and in other diseases was investigated by Western blot. Immunohistochemistry was performed on epiretinal membranes (ERM) obtained from ED patients to localize the 88 kDa protein. RESULTS: 88 kDa protein were detected in serum samples obtained from patients with posterior uveitis, tuberculosis, leprosy and rheumatoid arthritis. The 88 kDa protein found in serum from patients with ED is immunologically identical to that found in other systemic inflammatory conditions. 88 kDa protein was localized in inflammatory cells and in nonvascular endothelium in ERMs obtained from patients with ED. CONCLUSIONS: We have identified a novel acute phase reactant, which is elaborated in ocular and systemic inflammatory conditions other than Eales disease. Further work is necessary to decipher the precise role of the 88 kDa protein in the pathophysiology of these inflammatory diseases.

7523. San Gabriel P, Saiman L, Kaye K, Silin M, Onorato I, Schulte J. Completeness of pediatric TB reporting in New York City. Public Health Rep. 2003 Mar-Apr;118(2):144-53.

OBJECTIVE: Accurate surveillance of tuberculosis (TB) in children is critical because such cases represent recent transmission, but surveillance is difficult as only 10% to 50% of cases are culture-confirmed. Hospital-based sources were used to develop alternative surveillance to assess completeness of reporting for pediatric TB in northern Manhattan and Harlem from 1993 through 1995. METHODS: Alternative surveillance sources included ICD-9-CM hospital discharge codes for active TB and gastric aspirate reports. Cases identified by alternative surveillance were compared with cases previously reported to the New York City Department of Health (NYC DOH). RESULTS: Alternative surveillance detected 25 cases of possible pediatric TB, of which four (16%) had never been reported to the NYC DOH and three (12%) had been reported as suspect cases, but had not fulfilled the criteria for a reportable case of pediatric TB. Of these seven newly counted cases, three were detected by ICD-9-CM codes, three by a gastric aspirate log book, and one by both. In contrast, 13 other cases had been reported to the NYC DOH, but were undetected by our alternative surveillance; eight of these could be verified with available medical records. Thus, the demographic and clinical characteristics of the 25 detected and the eight undetected cases with available medical records were evaluated in this study. CONCLUSION: Alternative surveillance proved effective, was complementary to the NYC DOH surveillance efforts, and increased the number of pediatric TB cases identified during the study period by 21%.

7524. Soman D, Davies SJ. A suspected case of tuberculosis of the temporomandibular joint. Br Dent J. 2003 Jan 11;194(1):23-4.

Primary tuberculosis is symptomless in the great majority of individuals. The clinical appearance of tuberculosis (TB) of the temporomandibular joint is similar to that of ordinary arthritis of this joint and so, is unlikely to be diagnosed early in non-destructive disease, especially when there are no symptoms and signs of TB elsewhere in the body. The role of Mycobacterium tuberculosis and other opportunistic organisms should not be overlooked in complicated head and neck infections as this is an ever increasing problem today, especially due to the emergence of multi-drug resistant strains of tuberculosis in some urban areas.

7525. Stavri H, Moldovan O, Mihaltan F, Banica D, Doyle RJ. Rapid dot sputum and serum assay in pulmonary tuberculosis. J Microbiol Methods. 2003 Mar;52(3):285-96.

A rapid direct sputum (Sp.) and/or antibody assay, based on immunoblotting and enzyme immunoassay is described. The test can detect mycobacterial antigens or antibodies in clinical specimens from pulmonary tuberculosis (TB) patients. In this study, 87 sputa, 87 sera and 40 paired sputa and sera were utilized from smear-positive and smear-negative, culture-positive patients; 59 sputa, 37 sera and 22 paired sputa and sera from nontuberculosis respiratory disease patients and 68 sera from healthy controls. The antigen detection in sputum by dot assay has 86.1% sensitivity on active tuberculosis patients, 92.9% specificity, 91.6% positive predictive value (PPV), 88.2% negative predictive value (NPV) and 10.3% error. The antibody assay has 83.6% sensitivity, 95.4% specificity, 94.4% positive predictive value, 85.6% negative predictive value and 11% error. The test performed on paired sputum and serum (Sr.) samples has a sensitivity of 93.3%, which rose to 96.1% on smear-positive and culture-positive patients, but the specificity decreased to 83% in sputum, whereas in serum it was 92%. The results of the assay, combined with clinical and radiological data, could formthe basis for starting an earlier course of treatment for tuberculosis.

7526. Tailleux L, Schwartz O, Herrmann JL, Pivert E, Jackson M, Amara A, Legres L, Dreher D, Nicod LP, Gluckman JC, Lagrange PH, Gicquel B, Neyrolles O. DC-SIGN is the major Mycobacterium tuberculosis receptor on human dendritic cells. J Exp Med. 2003 Jan 6;197(1):121-7.

Early interactions between lung dendritic cells (LDCs) and Mycobacterium tuberculosis, the etiological agent of tuberculosis, are thought to be critical for mounting a protective anti-mycobacterial immune response and for determining the outcome of infection. However, these interactions are poorly understood, at least at the molecular level. Here we show that M. tuberculosis enters human monocyte-derived DCs after binding to the recently identified lectin DC-specific intercellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN). By contrast, complement receptor (CR)3 and mannose receptor (MR), which are the main M. tuberculosis receptors on macrophages (Mphis), appeared to play a minor role, if any, in mycobacterial binding to DCs. The mycobacteria-specific lipoglycan lipoarabinomannan (LAM) was identified as a key ligand of DC-SIGN. Freshly isolated human LDCs were found to express DC-SIGN, and M. tuberculosis-derived material was detected in CD14(-)HLA-DR(+)DC-SIGN(+) cells in lymph nodes (LNs) from patients with tuberculosis. Thus, as for human immunodeficiency virus (HIV), which is captured by the same receptor, DC-SIGN-mediated entry of M. tuberculosis in DCs in vivo is likely to influence bacterial persistence and host immunity.

7527. Underwood BR, White VL, Baker T, Law M, Moore-Gillon JC. Contact tracing and population screening for tuberculosis--who should be assessed? J Public Health Med. 2003 Mar;25(1):59-61.

BACKGROUND: The aim of the study was to investigate the relative effectiveness of four strategies in detecting and preventing tuberculosis: contact tracing of smear-positive pulmonary disease, of smear-negative pulmonary disease and of non-pulmonary disease, and screening new entrants. METHODS: An analysis of patient records and a TB database was carried out for an NHS Trust-based tuberculosis service in a socio-economically deprived area. Subjects were contacts of all patients treated for TB between 1997 and 1999. New entrants were screened in 1999. Outcomes measured were numbers of cases of active tuberculosis detected and numbers of those screened given chemoprophylaxis. RESULTS: A total of 643 contacts of 227 cases of active TB were seen, and 322 new entrants to the United Kingdom. The highest proportion of contacts requiring full treatment or chemoprophylaxis were contacts of smear-positive index cases (33 out of 263 contacts; 12.5 per cent). Tracing contacts of those with smear-negative pulmonary tuberculosis (12 out of 156; 7.7 per cent) and non-pulmonary disease (14 out of 277; 6.2 per cent) was significantly more effective in identifying individuals requiring intervention (full treatment or chemoprophylaxis) than routine screening of new entrants (10 out of 322; 3.1 per cent). CONCLUSIONS: Screening for TB of new entrants to the United Kingdom is part of the national programme for control and prevention of TB, whereas tracing contacts of those with smear-negative and non-pulmonary disease is not. This study demonstrates that, in our population, the contact-tracing strategy is more effective than new entrant screening. It is not likely that the contacts have caught their disease from the index case, but rather that in high-incidence areas such as ours such tracing selects extended families or communities at particularly high risk.

7528. Wang LJ, Wu CF, Wong YC, Chuang CK, Chu SH, Chen CJ. Imaging findings of urinary tuberculosis on excretory urography and computerized tomography. J Urol. 2003 Feb;169(2):524-8.

PURPOSE: We performed a retrospective study to analyze findings on excretory urography (IVP) and computerized tomography (CT) in patients with urinary tuberculosis. MATERIALS AND METHODS: In a 16-year period 53 patients with a total of 47 IVP and 33 CT examinations were diagnosed with urinary tuberculosis at our hospital. IVP and CT were reviewed and compared for certain imaging findings, including moth-eaten calices, renal parenchymal masses, an amputated infundibulum, autonephrectomy, thick urinary tract walls, urinary tract calcifications, renal parenchymal cavities, hydrocalycosis, hydronephrosis or hydroureter due to stricture, extra-urinary tubercular manifestations and renal parenchymal scarring. RESULTS: The most common finding on IVP was hydrocalycosis, hydronephrosis or hydroureter due to stricture, whereas renal parenchymal scarring was the most common finding on CT. Imaging findings of renal parenchymal masses and scarring, thick urinary tract walls and extra-urinary tubercular manifestations were significantly more common on CT than on IVP. Three imaging patterns were noted on all 44 IVPs (100%) and 31 of 33 CTs (94%) with multiple imaging findings, including multiple stricture sites, a single stricture with 1 other imaging finding and autonephrectomy with another imaging finding other than stricture. CONCLUSIONS: When the 3 imaging patterns are shown on IVP and CT, tubercular cultures or biopsies are suggested to make the definite diagnosis of urinary tuberculosis. Thus, treatment can be initiated as early as possible.

7529. Yilmaz E, Balci A, Sal S, Cakmakci H. Tuberculous ileitis in a renal transplant recipient with familial Mediterranean fever: Gray-scale and power Doppler sonographic findings. J Clin Ultrasound. 2003 Jan;31(1):51-4.

The ileocecal area is the most common site of involvement of intestinal tuberculosis. We report the case of a 26-year-old renal transplant recipient with familial Mediterranean fever who developed tuberculous ileitis. Gray-scale sonography and CT showed circumferential thickening of the bowel wall and enlargement of the mesenteric lymph nodes. Power Doppler sonography revealed markedly increased vascularity in the wall of the affected ileal segment and in the mesenteric nodes. Some nodes had no flow at the center owing to caseation necrosis, a finding consistent with the diagnosis of tuberculous ileitis. Colonoscopy was performed, and histopathologic examination of biopsy specimens revealed acute inflammatory changes. Cultures of the specimens confirmed the presence of Mycobacterium tuberculosis. We conclude that findings on power Doppler sonography may support a diagnosis of tuberculous ileitis and avoid clinical mismanagement. Copyright 2002 Wiley Periodicals, Inc.

7530. Zacharia TT, Shah JR, Patkar D, Kale H, Sindhwani V. MRI in ankle tuberculosis: Review of 14 cases. Australas Radiol. 2003 Mar;47(1):11-6.

We reviewed the MR imaging features of ankle tuberculosis and determined the role of MR in its diagnosis. A retrospective analysis of 14 cases of ankle tuberculosis imaged with MRI was performed. Plain radiographs were also reviewed where available, and the imaging characteristics were noted. We also reviewed the medical records in order to assess the impact of the imaging findings on management of these patients. Magnetic resonance imaging is extremely helpful for detection, mapping the extent and resolution of the disease. It can identify cases, enables early institution of antituberculous chemotherapy and might obviate the need for surgery.

7531. Zhou D, Lai X, Shen Y, Sehgal P, Shen L, Simon M, Qiu L, Huang D, Du GZ, Wang Q, Letvin NL, Chen ZW. Inhibition of adaptive Vgamma2Vdelta2+ T-cell responses during active mycobacterial coinfection of simian immunodeficiency virus SIVmac-infected monkeys. J Virol. 2003 Mar;77(5):2998-3006.

Adaptive immune responses of gammadelta T cells during active mycobacterial coinfection of human immunodeficiency virus-infected humans have not been studied. Macaques infected with the simian immunodeficiency virus (SIV) SIVmac were employed to determine the extent to which a coincident AIDS virus infection might compromise immune responses of mycobacterium-specific Vgamma2Vdelta2(+) T cells during active mycobacterial infection. Control SIVmac-negative macaques developed primary and recall expansions of phosphoantigen-specific Vgamma2Vdelta2(+) T cells after Mycobacterium bovis BCG infection and BCG reinfection, respectively. In contrast, SIVmac-infected macaques did not exhibit sound primary and recall expansions of Vgamma2Vdelta2(+) T cells in the blood and pulmonary alveoli following BCG infection and reinfection. The absence of adaptive Vgamma2Vdelta2(+) T-cell responses was associated with profound CD4(+) T-cell deficiency and subsequent development of SIVmac-related tuberculosis-like disease in the coinfected monkeys. Consistently, Vgamma2Vdelta2(+) T cells from coinfected monkeys displayed a reduced capacity to expand in vitro following stimulation with phosphoantigen. The reduced ability of Vgamma2Vdelta2(+) peripheral blood lymphocytes (PBL) to expand could be restored to some extent by coculture of these cells with CD4(+) T cells purified from PBL of SIV-negative monkeys. Furthermore, naive monkeys inoculated simultaneously with SIVmac and BCG were unable to sustain expansion of Vgamma2Vdelta2(+) T cells at the time that the coinfected monkeys developed lymphoid depletion and a fatal tuberculosis-like disease. Nevertheless, no deletion in Vdelta2 T-cell receptor repertoire was identified in SIVmac-BCG-coinfected macaques, implicating an SIVmac-induced down-regulation rather than a clonal exhaustion of these cells. Thus, an SIVmac-induced compromise of the adaptive Vgamma2Vdelta2(+) T-cell responses may contribute to the immunopathogenesis of the SIV-related tuberculosis-like disease in macaques.

Pathogenesis:

7532. Berrebi D, Bruscoli S, Cohen N, Foussat A, Migliorati G, Bouchet-Delbos L, Maillot MC, Portier A, Couderc J, Galanaud P, Peuchmaur M, Riccardi C, Emilie D. Synthesis of glucocorticoid-induced leucine zipper (GILZ) by macrophages: an anti-inflammatory and immunosuppressive mechanism shared by glucocorticoids and IL-10. Blood. 2003 Jan 15;101(2):729-38.

Glucocorticoids and interleukin 10 (IL-10) prevent macrophage activation. In murine lymphocytes, glucocorticoids induce expression of glucocorticoid-induced leucine zipper (GILZ), which prevents the nuclear factor kappaB (NF-kappaB)-mediated activation of transcription. We investigated whether GILZ could account for the deactivation of macrophages by glucocorticoids and IL-10. We found that GILZ was constitutively produced by macrophages in nonlymphoid tissues of humans and mice. Glucocorticoids and IL-10 stimulated the production of GILZ by macrophages both in vitro and in vivo. Transfection of the macrophagelike cell line THP-1 with the GILZ gene inhibited the expression of CD80 and CD86 and the production of the proinflammatory chemokines regulated on activation normal T-cell expressed and secreted (CCL5) and macrophage inflammatory protein 1alpha (CCL3). It also prevented toll-like receptor 2 production induced by lipopolysaccharide, interferongamma, or an anti-CD40 mAb, as well as NF-kappaB function. In THP-1 cells treated with glucocorticoids or IL-10, GILZ was associated with the p65 subunit of NF-kappaB. Activated macrophages in the granulomas of patients with Crohn disease or tuberculosis do not produce GILZ. In contrast, GILZ production persists in tumor-infiltrating macrophages in Burkitt lymphomas. Therefore, GILZ appears to play a key role in the anti-inflammatory and immunosuppressive effects of glucocorticoids and IL-10. Glucocorticoid treatment stimulates GILZ production, reproducing an effect of IL-10, a natural anti-inflammatory agent. The development of delayed-type hypersensitivity reactions is associated with the down-regulation of GILZ gene expression within lesions. In contrast, the persistence of GILZ gene expression in macrophages infiltrating Burkitt lymphomas may contribute tothe failure of the immune system to reject the tumor.

7533. Biswas P, Ferrarini M, Mantelli B, Fortis C, Poli G, Lazzarin A, Manfredi AA. Double-edged effect of Vgamma9/Vdelta2 T lymphocytes on viral expression in an in vitro model of HIV-1/mycobacteria co-infection. Eur J Immunol. 2003 Jan;33(1):252-63.

A reciprocal influence exists between mycobacteria and HIV: HIV-infected individuals are more susceptible to mycobacterial infections and, on the other hand, mycobacterial infection results inacceleration of HIV disease progression. Vgamma9/Vdelta2 T lymphocytes are known to participate in the defense against intracellular pathogens, including Mycobacterium tuberculosis. Indeed, they kill mycobacteria-infected macrophages and, upon recognition of mycobacterial Ag, release TNF-alpha and IFN-gamma, which are also up-regulators of HIV expression. To assess whether mycobacteria-activated gamma delta T lymphocytes contribute to the enhancement of HIV replication, we established an in vitro model mimicking HIV and mycobacteria co-infection with the latently HIV-infected promonocytic U1 cell line and Vgamma9/Vdelta2 peripheral lymphocytes stimulated with mycobacterial Ag. gamma delta T cell activation determined two distinct, but connected effects, namely U1cell death and HIV expression. Both effects were mainly mediated by release of TNF-alpha and IFN-gamma from activated gamma delta lymphocytes, although Fas-FasL interaction also contributed to U1 apoptosis. The final outcome on U1 survival, and thus, on HIV expression, highly depended on mycobacterial Ag concentration coupled to the differential secretory potency of gamma delta cells. In particular, the induction of viral expression prevailed at low Ag concentration and with lower cytokine production by mycobacteria-activated gamma delta cells. Notably, during the course of HIV infection, Vgamma9/Vdelta2 lymphocytes are reported to be functionally impaired and may thus indirectly influence the progression of HIV disease. In addition, a predominant inhibition of viral replication was encountered when mycobacteria-activated gamma delta T cells were co-cultured with primary HIV-infected macrophages. Thus, we suggest that specific recognition of mycobacterial Ag by gamma delta T lymphocytes in co-infected individuals may modulate viral replication through the complex array of soluble factors released.

7534. Brown RM, Cruz O, Brennan M, Gennaro ML, Schlesinger L, Skeiky YA, Hoft DF. Lipoarabinomannan-reactive human secretory immunoglobulin A responses induced by mucosal bacille Calmette-Guerin vaccination.J Infect Dis. 2003 Feb 1;187(3):513-7.

The ability of 17 recombinant mycobacterial proteins, native antigen 85 complex, lipoarabinomannan (LAM), and Mycobacterium tuberculosis lysate to detect antibody responses induced by bacille Calmette-Guerin (BCG) vaccination and active tuberculosis infection were studied in enzyme-linked immunosorbent assays. Only LAM-reactive serum immunoglobulin G responses were significantly increased in both BCG-vaccinated patients and patients with active tuberculosis (P<.05), and oral BCG vaccination also induced significant increases in LAM-reactive secretory immunoglobulin A (P<.05). LAM-reactive antibody assays can serve as markers of humoral and mucosal immunity in future trials of BCG and newer attenuated mycobacterial vaccines.

7535. Cheadle EJ, Selby PJ, Jackson AM. Mycobacterium bovis bacillus Calmette-Guerin-infected dendritic cells potently activate autologous T cells via a B7 and interleukin-12-dependent mechanism. Immunology. 2003 Jan;108(1):79-88.

Mycobacteria are potent adjuvants, can survive intracellularly and have been safely used for many years as vaccines against tuberculosis and leprosy. They are thus important potential vectors for recombinant vaccines. Many of their adjuvant properties are mediated following phagocytosis by dendritic cells (DC), which are in turn critical for priming naive T cells. Although the maturation of DC in response to mycobacteria, such as Mycobacterium bovis bacillus Calmette-Guerin (BCG), is well described the subsequent responses of autologous T cells to mycobacterium-infected DC remains uncharacterized. In our experiments DC infected with BCG expressed more co-stimulatory molecules than tumour-necrosis factor-alpha (TNF-alpha) -treated DC and stimulated more potent mixed leucocyte reactions. When autologous T cells were co-cultured with BCG-exposed DC they became highly activated, as determined by display of CD25, CD54 and CD71 on both CD4+ and CD8+ cells. In contrast, the response of T cells to TNF-alpha-matured DC was significantly less. Cytokine production from T cells cultured with BCG-exposed DC was enhanced with elevated secretion of interleukin-2 (IL-2), IL-10 and interferon-gamma (IFN-gamma) and was produced by both CD4+ and CD8+ lymphocytes as determined by intracellular staining. In particular, IFN-gamma secretion was increased from 50 pg/ml to 25 000 pg/ml and IL-10 secretion increased from 20 pg/ml to 300 pg/ml in BCG-exposed DC co-cultures. Blocking antibodies to B7.1 and B7.2 or IL-12 significantly reduced the secretion of IFN-gamma and reductions were also seen in the expression of CD25 and CD71 by CD4+ cells. These data demonstrate that mycobacterially infected DC are particularly potent activators of autologous T cells compared to TNF-alpha-exposed DC and that the resultant T cells are functionally superior.

7536. Cubillas-Tejeda AC, Ruiz-Arguelles A, Bernal-Fernandez G, Quiroz-Compean L, Lopez-Davila A, Reynaga-Hernandez E, Gonzalez-Amaro R. Cytokine production and expression of leucocyte-differentiation antigens by human mononuclear cells in response to mycobacterium tuberculosis antigens. Scand J Immunol. 2003 Feb;57(2):115-24.

The aim of this work was to characterize a leucocyte-differentiation antigen or chemokine receptor that allows the identification of type 1 (T helper 1 (Th1), Tc1) and type 2 (Th2, Tc2) lymphocytes in short-term-cultured human peripheral blood mononuclear cells. In addition, we assessed the type of response induced by mycobacterial antigens in tuberculosis patients and healthy contacts. Cells were stimulated with an unfractionated culture filtrate or 30 kDa antigen from Mycobacterium tuberculosis. Then, CD4 and CD8 cell labelling was combined with CD30, CD27, CD28, CD45RA or CD45R0 staining, detection of intracellular interferon-gamma (IFN-gamma) or interleukin-4 (IL-4) and analysis by three-colour flow cytometry. In separate experiments, the expression of different chemokine receptors (CCR1, CCR3, CCR5, CXCR3 and CXCR4) was also studied. We found that none of the cell-surface molecules studied was preferentially expressed by Th1 or Th2 cells. Thus, our results indicate that these lymphocyte subsets cannot be identified in short-term-cultured mononuclear cells on the basis of preferential expression of the cell markers studied, and that it is necessary to look for additional molecules that allow the discrimination of Th1 and Th2 cells.

7537. Day FH, Zhang Y, Clair P, Grabstein KH, Mazel M, Rees AR, Kaczorek M, Temsamani J. Induction of antigen-specific CTL responses using antigens conjugated to short peptide vectors. J Immunol. 2003 Feb 1;170(3):1498-503.

Linear peptides (SynB vectors) with specific sequence motifs have been identified that are capable of enhancing the transport of a wide range of molecules into cells. These peptide vectors have been used to deliver exogenous peptides and protein Ags across the cell membrane and into the cytoplasm of cells. Specifically, in vitro analysis indicated that these SynB peptides enhanced the uptake of two 9-mer peptide Ags, NP(147-155) and Mtb(250-258) (T cell epitopes of influenza nucleoprotein and Mycobacterium tuberculosis, respectively) and the M. tuberculosis Ag Mtb8.4 protein, into K562 cells when covalently linked to the respective Ags. Furthermore, selected SynB vectors, when conjugated to these same Ags and used as immunogens, resulted in considerably enhanced Ag-specific CTL responses. Several SynB vectors were tested and resulted in varying levels of cellular uptake. The efficiency of uptake correlated with the ability of the SynB construct to deliver each epitope in vivo and induce specific CTL responses in mice. These data suggest that peptide vectors, such as SynB that transport target Ags across the cell membrane in a highly efficient manner, have significant potential for vaccine delivery.

7538. Fletcher HA, Donoghue HD, Holton J, Pap I, Spigelman M. Widespread occurrence of Mycobacterium tuberculosis DNA from 18th-19th century Hungarians. Am J Phys Anthropol. 2003 Feb;120(2):144-52.

A large number (265) of burials from 1731-1838 were discovered in sealed crypts of the Dominican Church, Vac, Hungary in 1994. Many bodies were naturally mummified, so that both soft tissues and bones were available. Contemporary archives enabled the determination of age at death, and the identification of family groups. In some cases, symptoms before death were described and, occasionally, occupation. Initial radiological examination of a small number of individuals had indicated calcified lung lesions and demonstrable acid-fast bacteria suggestive of tuberculosis infection. Tuberculosis was endemic in 18th-19th century Europe, so human remains should contain detectable Mycobacterium tuberculosis complex (MTB) DNA, enabling comparisons with modern isolates. Therefore, a comprehensive examination of 168 individuals for the presence of MTB DNA was undertaken. Specific DNA amplification methods for MTB showed that 55% of individuals were positive and that the incidence varied according to age at death and sampling site in the body. Radiographs were obtained from 27 individuals and revealed an association between gross pathology and the presence of MTB DNA. There was an inverse relationship between PCR positivity and MTB target sequence size. In some cases, the preservation of MTB DNA was excellent, and several target gene sequences could be detected from the same sample. This information, combined with MTB DNA sequencing data and molecular typing techniques, will enable us to study the past epidemiology of TB infection, and extends the timeframe for studying changes in molecular fingerprints. Copyright 2003 Wiley-Liss, Inc.

7539. Fletcher HA, Donoghue HD, Taylor GM, van der Zanden AG, Spigelman M. Molecular analysis of Mycobacterium tuberculosis DNA from a family of 18th century Hungarians. Microbiology. 2003 Jan;149(Pt 1):143-51.

The naturally mummified remains of a mother and two daughters found in an 18th century Hungarian crypt were analysed, using multiple molecular genetic techniques to examine the epidemiology and evolution of tuberculosis. DNA was amplified from a number of targets on the Mycobacterium tuberculosis genome, including DNA from IS6110, gyrA, katG codon 463, oxyR, dnaA-dnaN, mtp40, plcD and the direct repeat (DR) region. The strains present in the mummified remains were identified as M. tuberculosis and not Mycobacterium bovis, from katG and gyrA genotyping, PCR from the oxyR and mtp40 loci, and spoligotyping. Spoligotyping divided the samples into two strain types, and screening for a deletion in the MT1801-plcD region initially divided the strains into three types. Further investigation showed, however, that an apparent deletion was due to poor DNA preservation. By comparing the effect of PCR target size on the yield of amplicon, a clear difference was shown between 18th century and modern M. tuberculosis DNA. A two-centre system was used to confirm the findings of this study, which clearly demonstrate the value of using molecular genetic techniques to study historical cases of tuberculosis and the care required in drawing conclusions. The genotyping and spoligotyping results are consistent with the most recent theory of the evolution and spread of the modern tuberculosis epidemic.

7540. Gardam MA, Keystone EC, Menzies R, Manners S, Skamene E, Long R, Vinh DC. Anti-tumour necrosis factor agents and tuberculosis risk: mechanisms of action and clinical management. Lancet Infect Dis. 2003 Mar;3(3):148-55.

Cases of active tuberculosis have been reported worldwide with the use of therapeutic agents that inhibit tumour necrosis factor (TNF) alpha. TNFalpha has a central role in mycobacterial infection and disease. Accordingly, progression of recently acquired tuberculosis infection or reactivation of remotely acquired infection should be expected with the use of anti-TNF agents. The available in-vitro and epidemiological evidence for the two currently approved agents, infliximab and etanercept, shows that the risk of development of active tuberculosis is greater with infliximab. Tuberculin skin testing (TST) should be undertaken before any significant immunosuppressive therapy including these agents, though the possibility of false-negative reactions in immunocompromised populations must be borne in mind. A positive TST should be followed by medical assessment and chest radiography, as well as by other tests judged appropriate by the physician to identify active disease. Active tuberculosis must be treated appropriately before initiation of treatment with an anti-TNF agent. Treatment of latent tuberculosis can be considered on an individual basis for TST-negative patients receiving anti-TNF agents when significant risk factors for infection are present.

7541. Gordon S. Alternative activation of macrophages. Nat Rev Immunol. 2003 Jan;3(1):23-35.

The classical pathway of interferon-gamma-dependent activation of macrophages by T helper 1 (T(H)1)-type responses is a well-established feature of cellular immunity to infection with intracellular pathogens, such as Mycobacterium tuberculosis and HIV. The concept of an alternative pathway of macrophage activation by the T(H)2-type cytokines interleukin-4 (IL-4) and IL-13 has gained credence in the past decade, to account for a distinctive macrophage phenotype that is consistent with a different role in humoral immunity and repair. In this review, I assess the evidence in favour of alternative macrophage activation in the light of macrophage heterogeneity, and define its limits and relevance to a range of immune and inflammatory conditions.

7542. Hwang HY, Chang CY, Chang LL, Chang SF, Chang YH, Chen YJ. Characterization of rifampicin-resistant Mycobacterium tuberculosis in Taiwan. J Med Microbiol. 2003 Mar;52(Pt 3):239-45.

Sixty-three rifampicin-resistant (Rif(r)) isolates of Mycobacterium tuberculosis from Kaohsiung, Taiwan, were analysed for mutations in the core region (69 bp, codons 511-533) of the rpoB gene. Some 84.1 % (53/63) of the resistant isolates showed mutations in this region, especially in codons 531 (41.5 %), 526 (18.9 %), 516 (15.1 %) and 533 (7.5 %). Five novel alleles of a total of 16 different types of mutations were identified in Rif(r) isolates. Ten Rif(r) isolates (15.9 %) exhibited no mutations in the core region of rpoB. Also, they did not show mutations in another 365 bp fragment (codons 99-220) of rpoB. The agar proportion method was used to determine the relationship between the degree of rifampicin resistance and alterations in the core region of rpoB. The results revealed that the mean MIC was 92.38 micro g ml(-1) for the 53 isolates with a mutation in the core region, whereas the mean MIC of the other 10 isolates without mutations was only 24.8 micro g ml(-1). This indicates that the isolates with mutations in the core region had higher levels of resistance than those without mutations in this region. IS6110 restriction fragment length polymorphism (RFLP) was used for typing of 55 Rif(r) M. tuberculosis isolates. Isolates contained two to 19 copies of IS6110, with sizes ranging from 600 to 16000 bp. The majority (85 %) contained six to 16 copies. No strains lacking IS6110 were found. A total of 54 of 55 RFLP types were defined at the 90 % similarity level. The observation of varied IS6110-associated banding patternsindicates that an outbreak of drug-resistant tuberculosis did not occur in this area.

7543. Jackson CJ, Lamb DC, Marczylo TH, Parker JE, Manning NL, Kelly DE, Kelly SL. Conservation and cloning of CYP51: a sterol 14 alpha-demethylase from Mycobacterium smegmatis. Biochem Biophys Res Commun. 2003 Feb 7;301(2):558-63.

The genetic locus encoding cytochrome P450 51 (CYP51; P450(14DM)) in Mycobacterium smegmatis is described here together with confirmation of activity in lanosterol 14 alpha-demethylation. The protein bound azole antifungals with high affinity and the rank order based on affinity matched the ranked order for microbiological sensitivity of the organism, thus supporting a possible role for CYP51 as a target in the antimycobacterial activity of these compounds. Non-saponifiable lipids were extracted from the bacteria grown on minimal medium. Unlike a previous report using growth on complex medium, no cholesterol was detected in two strains of M. smegmatis, but a novel lipid was detected. The genetic locus of CYP51 is discussed in relation to function; it is conserved as part of a putative operon in M. smegmatis, Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium bovis and consists of six open-reading frames including two CYPs and a ferredoxin under a putative Tet-R regulated promoter.

7544. Mayer K. Application of the protein truncation test (PTT) for the detection of tuberculosis sclerosis complex type 1 and 2 (TSC1 and TSC2) mutations. Methods Mol Biol. 2003;217:329-44. 65: No abstract available.

7545. Melmed G, Thomas LS, Lee N, Tesfay SY, Lukasek K, Michelsen KS, Zhou Y, Hu B, Arditi M, Abreu MT. Human intestinal epithelial cells are broadly unresponsive to Toll-like receptor 2-dependent bacterial ligands: implications for host-microbial interactions in the gut.J Immunol. 2003 Feb 1;170(3):1406-15.

Intestinal epithelial cells (IEC) interact with a high density of Gram-positive bacteria and are active participants in mucosal immune responses. Recognition of Gram-positive organisms by Toll-like receptor (TLR)2 induces proinflammatory gene expression by diverse cells. We hypothesized that IEC are unresponsive to Gram-positive pathogen-associated molecular patterns and sought to characterize the functional responses of IEC to TLR2-specific ligands. Human colonic epithelial cells isolated by laser capture microscopy and IEC lines (Caco-2, T84, HT-29) were analyzed for expression of TLR2, TLR6, TLR1, and Toll inhibitory protein (Tollip) mRNA by RT-PCR and quantitative real-time PCR. Response to Gram-positive bacterial ligands was measured by NF-kappa B reporter gene activation and IL-8 secretion. TLR2 protein expression was analyzed by immunofluorescence and flow cytometry. Colonic epithelial cells and lamina propria cells from both uninflamed and inflamed tissue demonstrate low expression of TLR2 mRNA compared with THP-1 monocytes. IECs were unresponsive to TLR2 ligands including the staphylococcal-derived Ags phenol soluble modulin, peptidoglycan, and lipotechoic acid and the mycobacterial-derived Ag soluble tuberculosis factor. Transgenic expression of TLR2 and TLR6 restored responsiveness to phenol soluble modulin and peptidoglycan in IEC. In addition to low levels of TLR2 protein expression, IEC also express high levels of the inhibitory molecule Tollip. We conclude that IEC are broadly unresponsive to TLR2 ligands secondary to deficient expression of TLR2 and TLR6. The relative absence of TLR2 protein expression by IEC and high level of Tollip expression may be important in preventing chronic proinflammatory cytokine secretion in response to commensal Gram-positive bacteria in the gut.

7546. Mirdha BR. Mycobacterium avium-intracellulare in stool in HIV-seropositive man. Indian J Gastroenterol. 2003 Jan-Feb;22(1):25.

Mycobacterium tuberculosis is the predominant acid-fast bacillus causing diarrhea in HIV-seropositive patients in India. We report a 27-year-old HIV-seropositive man with diarrhea in whom M. avium-intracellulare was isolated on stool culture.

7547. Riendeau CJ, Kornfeld H. THP-1 cell apoptosis in response to Mycobacterial infection. Infect Immun. 2003 Jan;71(1):254-9.

We previously reported that Mycobacterium tuberculosis infection primes human alveolar macrophages (HAM) for tumor necrosis factor alpha (TNF-alpha)-mediated apoptosis and that macrophage apoptosis is associated with killing internalized bacilli. Virulent mycobacterial strains elicit much less apoptosis than attenuated strains, implying that apoptosis is a defense against intracellular infection. The present study evaluated the potential for phorbol myristate acetate-differentiated THP-1 cells to mimic this response of primary macrophages. Consistent with the behavior of alveolar macrophages, attenuated M. tuberculosis H37Ra and Mycobacterium bovis BCG strongly induce THP-1 apoptosis, which requires endogenous TNF. THP-1 apoptosis is associated with reduced viability of infecting BCG. In contrast, virulent wild-type M. tuberculosis H37Rv and M. bovis do not increase THP-1 apoptosis over baseline. BCG induced early activation of caspase 10 and 9, followed by caspase 3. In contrast, wild-type M. bovis infection failed to activate any caspases in THP-1 cells. BCG-induced THP-1 apoptosis is blocked by retroviral transduction with vectors expressing crmA but not bcl-2. We conclude that differentiated THP-1 cells faithfully model the apoptosis response of HAM. Analysis of the THP-1 cell response to infection with virulent mycobacteria suggests that TNF death signals are blocked proximal to initiator caspase activation, at the level of TNF receptor 1 or its associated intracytoplasmic adaptor complex. Interference with TNF death signaling may be a virulence mechanism that allows M. tuberculosis to circumvent innate defenses leading to apoptosis of infected host cells.

7548. Sese E, Xiol X, Castellote J, Rodriguez-Farinas E, Tremosa G. Low complement levels and opsonic activity in hepatic hydrothorax: its relationship with spontaneous bacterial empyema. J Clin Gastroenterol. 2003 Jan;36(1):75-7.

GOALS: To analyze the pleural fluid factors that might cause spontaneous bacterial empyema (SBEM) in patients with cirrhotic hydrothorax. BACKGROUND: Pathogenic mechanism of SBEM of cirrhotic patients is probably similar to that of spontaneous bacterial peritonitis, but local factors affecting pleural fluid have not been studied. STUDY: Determination of C3, C4, and opsonic activity levels of pleural fluid in a cohort of patients with pleural effusions of different causes. RESULTS: Forty-eight patients had hepatic hydrothorax; 8, heart failure and 45, exudates (9, tuberculosis; 21, malignancies; 10, other). Of the 48 cirrhotic patients, 15 developed SBEM on admission. The pleural fluid of cirrhotic patients showed significantly lower levels of total protein, complement, and opsonic activity than did the fluids of patients with other causes of SBEM. Patients who developed SBEM had lower concentrations of pleural fluid total protein and C3 and had a higher Child-Pugh score than patients who did not develop the infection. CONCLUSION: Cirrhotic patients with hepatic hydrothorax have lower pleural fluid opsonic activity and C3 levels than those found in the pleural fluid of patients with other causes. Patients who develop SBEM have lower levels of pleural fluid C3, pleural fluid total protein, and a higher Child-Pugh score than those who do not develop SBEM.

7549. Sieling PA, Chung W, Duong BT, Godowski PJ, Modlin RL. Toll-like receptor 2 ligands as adjuvants for human Th1 responses. J Immunol. 2003 Jan 1;170(1):194-200.

Bacterial lipopeptides (bLPs) are increasingly used as adjuvants to activate cell-mediated immune responses to foreign Ags. To explore mechanisms whereby bLPs adjuvant T cell responses, we stimulated human PBMCs with bLPs. We found that bLPs stimulate T cells to proliferate and produce IFN-gamma in an accessory cell-dependent manner and in the absence of exogenous protein Ags. The ability of bLPs to stimulate T cell proliferation was Toll-like receptor 2 dependent and required IL-12, interaction with costimulatory molecules, and MHC proteins. Our data suggest that bLPs adjuvant adaptive Th1 responses by enhancing Agpresentation of endogenous peptides.

7550. Sly LM, Hingley-Wilson SM, Reiner NE, McMaster WR. Survival of Mycobacterium tuberculosis in host macrophages involves resistance to apoptosis dependent upon induction of antiapoptotic Bcl-2 family member Mcl-1.J Immunol. 2003 Jan 1;170(1):430-7.

Mcl-1 protein expression was found to be up-regulated during infection with virulent Mycobacterium tuberculosis strain H37Rv. Mcl-1 induction in THP-1 cells was optimal at a multiplicity of infection of 0.8-1.2 bacilli per macrophage and was independent of opsonin coating of the bacteria. Mcl-1 expression was elevated as early as 4 h, peaked at 5.8-fold above control cells at 24 h, and remained elevated at 48 h after infection. In THP-1 cells, mMcl-1 mRNA was induced by infection with live H37Rv but not with attenuated M. tuberculosis strain H37Ra, heat-killed H37Rv, or latex beads. In THP-1 cells and monocyte-derived macrophages (MDMs), Mcl-1 protein was induced by infection with live H37Rv but not with attenuated M. tuberculosis strain H37Ra, heat-killed H37Rv, or latex beads. Treatment of uninfected, H37Ra-infected, and H37Rv-infected THP-1 cells and MDMs with antisense oligonucleotides to mcl-1 reduced Mcl-1 expression by >84%. This resulted in an increase in apoptosis of both MDMs and THP-1 cells that were infected with H37Rv, but not cells that were uninfected or infected with H37Ra. Increased apoptosis correlated with a decrease in M. tuberculosis CFUs recovered from antisense-treated, H37Rv-infected cells at 4 and 7 days after infection. In contrast, CFU recoveries from sense-treated, H37Rv-infected cells or from antisense- or sense-treated, H37Ra-infected cells were unchanged from controls. Thus, the antiapoptotic effect of the induction of Mcl-1 expression in H37Rv-infected macrophages promotes the survival of virulent M. tuberculosis.

7551. Vankayalapati R, Wizel B, Weis SE, Klucar P, Shams H, Samten B, Barnes PF. Serum cytokine concentrations do not parallel Mycobacterium tuberculosis-induced cytokine production in patients with tuberculosis. Clin Infect Dis. 2003 Jan 1;36(1):24-8.

We measured serum cytokine concentrations and Mycobacterium tuberculosis-stimulated cytokine production by peripheral blood mononuclear cells (PBMCs) obtained from persons infected with M. tuberculosis. Serum interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) concentrations were elevated in patients with tuberculosis compared with healthy persons who had reactions to tuberculin skin tests, but IL-18 concentrations were not. In contrast, M. tuberculosis-stimulated PBMCs from patients with tuberculosis produced less IFN-gamma and IL-18 but similar amounts of IL-10, compared with PBMCs from healthy subjects who had reactions to tuberculin skin tests. Pretreatment of PBMCs from healthy subjects with reaction to tuberculin with serum from patients with tuberculosis inhibited IFN-gamma production in response to M. tuberculosis, and inhibition was blocked by anti-IL-10. Thus, serum concentrations of IFN-gamma, IL-18, and IL-10 do not parallel M. tuberculosis-induced cytokine levels, and increased IL-10 serum levels in patients with tuberculosis inhibit IFN-gamma production in response to mycobacterial antigens.

Therapy:

7552. Hollender ES, Ashkin D, Narita M. Urine color test to monitor isoniazid compliance: "pissin' in the wind"? Chest. 2003 Mar;123(3):668-70. No abstract available

7553. Saracli MA, Albay A, Guney C, Alpt A, Ciftci F, Doganci L. Resistance patterns of Mycobacterium tuberculosis complex isolates in the Turkish Army from 1998 to 2000. Mil Med. 2003 Jan;168(1):24-7.

The aim of this retrospective study was to investigate susceptibility rates of Mycobacterium tuberculosis complex (MTBC) isolates against streptomycin, rifampicin, isoniazid, and ethambutol between January 1998 and December 2000 in the Turkish Army. Specimens collected from patients were cultured both conventionally and radiometrically. Differentiation of MTBC bacteria from Mycobacteria other than tuberculosis bacilli was made by the BACTEC p-nitro-alpha-acetyl-amino-beta-hydroxypropiophenone test. Susceptibility testing of MTBC isolates was performed using the BACTEC radiometric susceptibility assay for mycobacteria. Most of the specimens originated from respiratory system. A total of 98 isolates in 1998, 123 isolates in 1999, and 84 isolates in 2000 were obtained and identified as MTBC using the radiometric BACTEC TB460 system. Initial resistance was most frequent to isoniazid followed by ethambutol, streptomycin , and rifampicin in this study period. The differences between resistance rates were not statistically significant on an annual basis. None of these isolates was resistant to all four antimycobacterial agents. Although resistance rates of our isolates were not as high as previously reported by some authors from Turkey and there was no significant difference between the annual susceptibility rates, routine screening of antituberculosis drug susceptibility should be continued to control the resistance development and its spread.

7554. Schafer MP, Martinez KF, Mathews ES. Rapid detection and determination of the aerodynamic size range of airborne mycobacteria associated with whirlpools. Appl Occup Environ Hyg. 2003 Jan;18(1):41-50.

Novel environmental air and water mycobacteria sampling and analytical methods are needed to circumvent difficulties associated with the use of culture-based methodologies. To implement this objective, a commercial, clinical, genus DNA amplification method utilizing the polymerase chain reaction (PCR) was interfaced with novel air sampling strategies in the laboratory. Two types of air samplers, a three-piece plastic, disposable filter cassette and an eight-stage micro-orifice uniform deposit impactor (MOUDI), were used in these studies. In both samplers, 37-mm polytetrafluoroethylene (PTFE) filters were used. Use of the MOUDI sampler permitted the capture of airborne mycobacteria in discrete size ranges, an important parameter for relating the airborne mycobacteria cells to potential respirable particles (aerodynamic diameter <10 microm) capable of causing health effects. Analysis of the samples was rapid, requiring only 1-1.5 days, as no microbial culturing or DNA purification was required. This approach was then used to detect suspected mycobacteria contamination associated with pools at a large public facility. PCR was also used to analyze various water samples from these pools. Again, no culturing or sample purification was required. Water samples taken from all ultraviolet light/hydrogen peroxide-treated whirlpools tested positive for the presence of mycobacteria. No mycobacteria were detected in the chlorine-treated pools and the water main supply facility. All air samples collected in the proximity of the indoor whirlpools and the associated changing rooms were strongly positive for airborne mycobacteria. The airborne mycobacteria particles were predominantly collected on MOUDI stages 1-6 representing an aerodynamic size range of 0.5 to 9.9 microm. In conclusion, using this approach permits the rapid detection of mycobacteria contamination as well as the routine monitoring of suspected pools. The approach circumvents problems associated with culture-based methods such as fungal overgrowth on agar plates, and the presence of nonculturable or difficult to culture mycobacteria strains.

October 2003

8167. Agadi S. Diagnosis of tuberculosis. Lancet. 2003 Jun 14;361(9374):2082; author reply 2082-3. No abstract.

8168. Ahmed N, Caviedes L, Alam M, Rao KR, Sangal V, Sheen P, Gilman RH, Hasnain SE. Distinctiveness of Mycobacterium tuberculosis genotypes from human immunodeficiency virus type 1-seropositive and -seronegative patients in Lima, Peru. J Clin Microbiol. 2003 Apr;41(4):1712-6.

Genotypic analysis of Mycobacterium tuberculosis isolates obtained from human immunodeficiency virus type 1 (HIV-1)-seropositive (n = 80) and -seronegative (n = 25) patients from Lima, Peru, revealed two distinct genotypes correlating with the host immune status. While the level of intrastrain diversity of DNA fingerprints of HIV-seropositive isolates was less pronounced, these isolates showed many clonal groupings.

8169. Banerji S, Gupta S, Shende N, Satish K & Harinath BC. Serodiagnosis of tuberculosis using two ELISA systems. Indian J. of Clin. Biochem 2003, 18(2): 48-53.

8170. Bilen CY, Inci K, Erkan I, Ozen H. The predictive value of purified protein derivative results on complications and prognosis in patients with bladder cancer treated with bacillus Calmette-Guerin. J Urol. 2003 May;169(5):1702-5.

PURPOSE: We investigate the correlation of purified protein derivative (PPD) results before intravesical bacillus Calmette-Guerin (BCG) instillations with prognosis and complications of BCG. MATERIALS AND METHODS: A total of 57 men and 4 women with proven intermediate or high risk superficial bladder cancer received 6 courses of intravesical BCG instillations following complete resection of tumors. Skin reactivity to a PPD derivative of Mycobacterium tuberculosis was tested before starting and 1 week after BCG. The test was considered positive if the induration was 10 mm. or more in diameter after 48 or 72 hours. The patients were grouped according to PPD responses and symptoms. The statistical analyses were performed between PPD positive and negative groups, and also between symptomatic and asymptomatic patients. The groups were compared for relapse rates, time to first recurrence, complication rates and clinical outcome. RESULTS: Most of the patients with systemic side effects were in the PPD positive group but only fever had a statistically significant difference and was more frequent in the positive group (p <0.05). The recurrence-free intervals after intravesical BCG therapy did not differ significantly between PPD positive and negative groups. However, the trend of longer recurrence-free survival was evident for symptomatic patients (p = 0.056). The numbers of tumor recurrences were 10 (52%) in the PPD negative group and 19 (51%) in the PPD positive group, which was statistically insignificant. CONCLUSIONS: Patients with systemic reactions to BCG had the longest disease-free survival. It seems that patients with an augmented reaction to BCG probably have better antitumor activity. Furthermore, although larger groups of patients are mandatory for statistical analysis, this study shows that hypersensitivity reaction against tuberculin could alert physicians of severe complications.

8171. Bhatia AS, Satish K & Harinath BC. Immunodiagnosis of tuberculosis: An update. Indian J. of Clin Biochem 2003; 18 (2): 1-5.

8172. Brun V, Revel MP, Danel C, Fournier LS, Souilamas R, Frija G. Case report. Pyothorax-associated lymphoma: Diagnosis at percutaneous core biopsy with CT guidance. AJR Am J Roentgenol. 2003 Apr;180(4):969-71. No abstract.

8173. Candela A, Andujar J, Hernandez L, Martin C, Barroso E, Arriero JM, Romero S. Functional sequelae of tuberculous pleurisy in patients correctly treated. Chest. 2003 Jun;123(6):1996-2000.

STUDY OBJECTIVES: To assess the functional sequelae (FS) of patients with tuberculous pleurisy (TP), to analyze the influence of different factors in the occurrence of these FS, and, finally, to evaluate the relationship between the FS and roentgenographic sequelae. DESIGN: An observational, retrospective study. SETTING: A community teaching hospital in Alicante, Spain. PATIENTS AND METHODS: From April 1986 to July 2000, all patients with a firmly established diagnosis of TP, who had been functionally studied at the end of follow-up, were included in the study. A diagnosis of TP was considered to be definitive when the presence of granuloma on a pleural biopsy specimen was demonstrated or when a culture was positive for Mycobacterium tuberculosis in pleural fluid (PF) or tissue. The general characteristics of the study population and PF were compared in patients with or without restrictive FS (ie, FVC or TLC < 80%), looking for risk factors for developing this complication. RESULTS: Eighty-one of 150 patients who had been treated for TP were eligible for the study. At the end of follow-up, eight patients (10%) had a restrictive FS. These patients had a lower PF lactate dehydrogenase concentration (p < 0.001), a higher PF concentration of cholesterol (p < 0.03) and triglycerides (p < 0.03), and a higher percentage of lymphocytes (p < 0.04). A weak correlation was found between the FVC and the intensity of radiographic pleural thickening (r = - 0.298; p < 0.01). CONCLUSIONS: The FS in patients with TP is restrictive in type, infrequent, and usually mild. A higher PF lipid content or a more chronic inflammatory pleural reaction at diagnosis appear to be risk factors for developing a FS. The correlation between FS and roentgenographic sequelae is poor.

8174. Cataloluk O, Cakmak EA, Buyukberger N, Barlas O. Formalin fixing and paraffin embedding may lead to extra band development in PCR. New Microbiol. 2003 Apr;26(2):193-8.

The molecular biological analysis of infectious agents requires the availability of a reliable source of microorganisms to be used to recover DNA. Clinical samples can be obtained directly from infected patients or can be propagated using in vitro or in vivo systems. However, repeated sampling from patients is not always possible as the procedure may be invasive or unpleasant, or it is not possible to catch the same agent at the time of second sampling. Moreover, the techniques used may also produce false-positive and false-negative results. We therefore studied the impact of formalin-fixing and paraffin embedding on tissue sampling, and the methodologies such as DNA isolation and PCR amplification of DNAs from archival materials in the diagnosis of Mycobacterium tuberculosis. PCR analyses were done according to standard methods with some modifications. Demonstration of mycobacteria was successful both in tissue sections of the formalin-fixed lymph nodes and in stained fresh materials from patients. However, the results showed the presence of two extra bands in the gel. We accounted for extra band development due to the harshness of the methodology used to isolate nucleic acids from formalin-fixed and paraffin embedded tissue samples or the nature of the fixation procedure, or because of the time passed during storage in which alteration in the chromosomal DNA would take place. Thus, if disease- and tissue specific morphological features, such as sample size, type of fixation, and intralesional heterogeneity are ignored, errors because of sampling and methodologies used may lead to false-positive and false-negative results.

8175. Chow KM, Chow VC, Szeto CC. Indication for peritoneal biopsy in tuberculous peritonitis. Am J Surg. 2003 Jun;185(6):567-73. Review.

BACKGROUND: With the introduction of effective antituberculous chemotherapy, the clinical outcome of tuberculous peritonitis depends much on the diagnostic accuracy of this disease entity. This review summarizes the current state-of-the-art thinking regarding the protean manifestation and diagnostic modalities of this major infectious disease. DATA SOURCES: This review was compiled after an extensive search of the current and historical literature, comprising 1,070 cases of tuberculous peritonitis. A number of important areas were highlighted, with emphasis on the diagnostic value and clinical impact of peritoneal biopsy. CONCLUSIONS: We believe an aggressive diagnostic approach, particularly with peritoneal biopsy, is warranted for the diagnosis and timely treatment of tuberculous peritonitis.

8176. Cobos-Marin L, Montes-Vargas J, Rivera-Gutierrez S, Licea-Navarro A, Gonzalez-y-Merchand JA, Estrada-Garcia I. A novel multiplex-PCR for the rapid identification of Mycobacterium bovis in clinical isolates of both veterinary and human origin. Epidemiol Infect. 2003 Jun;130(3):485-90.

Bovine tuberculosis is a zoonotic disease that not only causes huge economic losses but also poses an important risk for human infection. The definitive identification of a clinical isolate relies on time-consuming, highly specialized and laborious biochemical tests. We have developed a method for the rapid and reliable identification of Mycobacterium bovis and for its simultaneous differentiation from other members of the M. tuberculosis complex. Furthermore, the technique also allowed us to distinguish M. tuberculosis complex members from other Mycobacterial species. The method comprises both a single PCR and a multiplex-PCR and can be confidently applied to samples of both veterinary and human origin.

8177. Cohen-Gadol AA, Zikel OM, Miller GM, Aksamit AJ, Scheithauer BW, Krauss WE. Spinal cord biopsy: a review of 38 cases. Neurosurgery. 2003 Apr;52(4):806-15; discussion 815-6.

OBJECTIVE: Neurosurgeons are frequently asked to evaluate patients for spinal cord biopsies when preoperative magnetic resonance imaging studies demonstrate nonspecific features. These lesions often appear unresectable, but surgeons must decide whether a biopsy is warranted. To determine the best approach to these cases, we evaluated the clinicopathological findings for patients with unknown spinal cord lesions who underwent spinal cord biopsies. METHODS: Thirty-eight consecutive patients who underwent spinal cord biopsies at the Mayo Clinic (Rochester, MN) between August 1988 and July 1998 were studied. A detailed review of the case histories, radiological results, surgical notes, histological findings, and outcomes was performed. RESULTS: Spinal cord biopsies were performed for 21 male and 17 female patients (mean age, 42.1 yr) with progressive neurological deficits related to spinal cord lesions. All patients underwent preoperative magnetic resonance imaging evaluations. High T2-weighted signal intensity and spinal cord expansion were identified in 92 and 87% of cases, respectively. After gadolinium infusion, the majority (94%) of the inflammatory lesions demonstrated patchy and often peripherally situated enhancement. This neuroradiological pattern was less common for neoplasms (50%) and benign lesions (40%). The most common pathological findings were inflammatory changes of demyelination or sarcoidosis, which together accounted for 13 cases (34%). Nonspecific changes or benign lesions were observed in 10 cases (26%). Neoplasms were identified in eight cases (21%). One case of tuberculosis and one of schistosomiasis were found. Overall, 47% of the preoperative diagnoses made by the attending surgeon were correct. For 26% of the patients, specific treatment was based on the biopsy results. The average follow-up period was 12 months (standard deviation, 14 mo; range, 0-50 mo). CONCLUSION: Preoperative laboratory and imaging studies are often diagnostically inconclusive in cases of spinal cord lesions with nonspecific features. Biopsies should be considered for patients with progressive symptomatic lesions.

8178. Drancourt M, Carrieri P, Gevaudan MJ, Raoult D. Blood agar and Mycobacterium tuberculosis: the end of a dogma. J Clin Microbiol. 2003 Apr;41(4):1710-1.

Incidental blood agar-based recovery of Mycobacterium tuberculosis led us to further investigate this routine medium for primary isolation and culture of M. tuberculosis. Fifteen respiratory tract and eight lymph node Ziehl-Neelsen-positive specimens were inoculated in parallel into tubes containing egg-based medium and 5% sheep blood agar. Colonies appeared sooner on this medium than on the egg-based medium, but this difference was not significant (P = 0.11, analysis of variance [ANOVA] test). Further experiments compared the growth of 38 respiratory and lymph node M. tuberculosis isolates when subcultured on the two media. After 6 days of incubation, 21 of 38 isolates had grown on blood agar, and the mean number of colonies was significantly greater on blood agar than on the egg-based medium (P < 0 0.001, ANOVA test). These results demonstrate that M. tuberculosis grows easily on blood agar within 1to 2 weeks, indicating that this basic medium is suitable for laboratory diagnosis of tuberculosis in addition to other media. Laboratories that routinely use prolonged incubations of blood plates, for example, for the recovery of Bartonella species, should consider the potential safety implications of encountering this highly infectious pathogen.

8179. Ewer K, Deeks J, Alvarez L, Bryant G, Waller S, Andersen P, Monk P, Lalvani A. Comparison of T-cell-based assay with tuberculin skin test for diagnosis of Mycobacterium tuberculosis infection in a school tuberculosis outbreak. Lancet. 2003 Apr 5;361(9364):1168-73.

BACKGROUND: The diagnosis of latent tuberculosis infection relies on the tuberculin skin test (TST), which has many drawbacks. However, to find out whether new tests are better than TST is difficult because of the lack of a gold standard test for latent infection. We developed and assessed a sensitive enzyme-linked immunospot (ELISPOT) assay to detect T cells specific for Mycobacterium tuberculosis antigens that are absent from Mycobacterium bovis BCG and most environmental mycobacteria. We postulated that if the ELISPOT is a more accurate test of latent infection than TST, it should correlate better with degree of exposure to M tuberculosis. METHODS: A large tuberculosis outbreak in a UK school resulted from one infectious index case. We tested 535 students for M tuberculosis infection with TST and ELISPOT. We compared the correlation of these tests with degree of exposure to the index case and BCG vaccination. FINDINGS: Although agreement between the tests was high (89% concordance,kappa=0.72, p<0.0001), ELISPOT correlated significantly more closely with M tuberculosis exposure than did TST on the basis of measures of proximity (p=0.03) and duration of exposure (p=0.007) to the index case. TST was significantly more likely to be positive in BCG-vaccinated than in non vaccinated students (p=0.002), whereas ELISPOT results were not associated with BCG vaccination (p=0.44). INTERPRETATION: ELISPOT offers a more accurate approach than TST for identification of individuals who have latent tuberculosis infection and could improve tuberculosis control by more precise targeting of preventive treatment.

8180. Fisk TL, Hon HM, Lennox JL, Fordham von Reyn C, Horsburgh CR Jr. Detection of latent tuberculosis among HIV-infected patients after initiation of highly active antiretroviral therapy. AIDS. 2003 May 2;17(7):1102-4.

This cross-sectional study of 110 individuals examined skin testing for latent tuberculosis infection (LTBI) after the initiation of highly active antiretroviral therapy. Skin test reactivity to one or more of four antigens was found in 98 out of 110 subjects (89%), and was maximal in those whose CD4 cell counts recovered to >= 100 cells/mm3. Skin testing is reliable for the identification or exclusion of LTBI once the CD4 cell count recovers to >= 100 cells/mm3.

8181. Hoskyns W. Paediatric tuberculosis. Postgrad Med J. 2003 May;79(931):272-8. Review.

Children are important in the epidemiology of tuberculosis as a marker of recent disease transmission and a reservoir for the future. Once infected they have a higher risk of progressing to tuberculous disease. Chest radiography and tuberculin testing with or without tissue for culture are still the standard tools for confirming the diagnosis once this is considered. Well researched treatment protocols are available but multidrug resistant tuberculosis and coexistent HIV are a challenge. Ensuring compliance with treatment is a major concern. Controversy still surrounds the place of BCG. Advances in the molecular genetics of tuberculosis hold out the possibility of better vaccines.

8182. Jain A, Verma RK, Tiwari V, Goel MM. Development of a new antigen detection dot-ELISA for diagnosis of tubercular lymphadenitis in fine needle aspirates. J Microbiol Methods. 2003 Apr;53(1):107-12.

A sandwich dot enzyme-linked immunosorbent assay (ELISA) was standardized to detect mycobacterial antigen in fine needle aspirates of patients with tubercular lymphadenitis (TBLN). The assay was performed on nitrocellulose paper by using antibodies raised in mice and rabbits against crude soluble protein (CSP) of Mycobacterium tuberculosis. The test was able to detect as low as 5 ng protein/ml. A total of 225 suspected cases of tubercular lymphadenopathy were screened, out of which 96 were cytomorphologically confirmed as cases of tubercular lymphadenitis (50 acid-fast bacilli (AFB)-positive and 46 AFB-negative). These were considered as positive controls. Only 28 cases were proven to be of nontubercular etiology and were considered as negative controls. In the remaining 101 (39 scanty) aspirates, tubercular etiology could neither be ruled out nor confirmed. Out of 50 AFB-positive confirmed cases of tubercular lymphadenitis, 46 were ELISA-positive. Out of 46 AFB-negative but cytomorphologically confirmed aspirates, antigen could be demonstrated in only 42 aspirates. Four samples from patients with nontubercular etiology were also found to be ELISA-positive. Antigen was picked up in a total of 90.3% of aspirates with suspicion of tuberculosis and 79.5% of scanty aspirates. The assay was found to be 91.6% sensitive and 85.7% specific. The assay was found to be simple and rapid, and hence, could be performed in areas where health facilities are rudimentary.

8183. Janmeja AK; Das SK; Kochhar S; Handa U. Tuberculosis of the parotid gland. Indian Journal of Chest Diseases and Allied Sciences. 2003 Jan-Mar; 45(1): 67-9.

Tuberculosis of the parotid gland is a rare entity. Only about a hundred cases have been reported till date, mostly from parotidectomy specimens. The present case was diagnosed by fine needle aspiration and treated successfully by short-course antitubercular chemotherapy. An early diagnosis can avoid parotidectomy, which can be a hazardous procedure in a medically treatable condition.

8184. Kerleguer A, Koeck JL, Fabre M, Gerome P, Teyssou R, Herve V. Use of equivocal zone in interpretation of results of the amplified mycobacterium tuberculosis direct test for diagnosis of tuberculosis. J Clin Microbiol. 2003 Apr;41(4):1783-4.

The sensitivity and specificity of the Amplified Mycobacterium Tuberculosis Direct (AMTD) test, evaluated with 1,363 respiratory samples (128 from tuberculous patients), were 92.97 and 98.7%, respectively. When an equivocal zone (30,000 to 1,000,000 relative light units [RLU]) was used instead of a 30,000-RLU cutoff, the sensitivity and specificity of the AMTD test were 92.97 and 100%, respectively.

8185. Kim SJ, Seok JW, Kim IJ, Kim YK, Kim DS. Multifocal Pott's disease (tuberculous spondylitis) incidentally detected on Tc-99m MDP bone and Ga-67 citrate scintigraphy in a patient with diabetes. Clin Nucl Med. 2003 Apr;28(4):286-9.

Pott's disease is an uncommon extrapulmonary form of tuberculosis. Delay in diagnosis and management may cause serious complications. The authors describe Pott's disease incidentally detected on Tc-99m MDP bone and Ga-67 imaging in a patient with diabetes. Tc-99m MDP bone scintigraphy showed intensely increased uptake in the lower cervical spine and lumbosacral regions. Ga-67 scintigraphy revealed intensely increased uptake corresponding to the areas noted on Tc-99m MDP bone scintigraphy. Magnetic resonance imaging showed destructive lesions in the C5-C6 and L5-S1 intervertebral discs with destruction of adjacent end plates. Biopsy of the lumbosacral area was guided by computed tomography, and histologic examination of the bone specimen showed caseation, giant cells, and acid-fast bacilli. Posterior decompression and posterolateral spinal fusion with bone grafts were performed. Antituberculous chemotherapy with isoniazid, rifampicin, pyrazinamide, and ethambutol was started. The patient showed remarkable relief of symptoms during a period of 9 months of therapy. Both Tc-99m MDP bone and Ga-67 imaging can offer the convenience of screening the entire body to detect multiple sites of Pott's disease.

8186. Koguchi Y, Kawakami K, Uezu K, Fukushima K, Kon S, Maeda M, Nakamoto A, Owan I, Kuba M, Kudeken N, Azuma M, Yara S, Shinzato T, Higa F, Tateyama M, Kadota J, Mukae H, Kohno S, Uede T, Saito A. High plasma osteopontin level and its relationship with interleukin-12-mediated type 1 T helper cell response in tuberculosis. Am J Respir Crit Care Med. 2003 May 15;167(10):1355-9. Epub 2003 Feb 05.

Osteopontin (OPN, also known as Eta-1), a noncollagenous matrix protein produced by macrophages and T lymphocytes, is expressed in granulomatous lesions caused by Mycobacterium tuberculosis infection. In the present study, we compared plasma concentrations of OPN in patients with active pulmonary tuberculosis with those of healthy control subjects and patients with sarcoidosis, another disease associated with granuloma formation. Plasma OPN levels were significantly higher in patients with tuberculosis (n = 48) than in control subjects (n = 34) and patients with sarcoidosis (n = 20). OPN levels correlated well with severity of pulmonary tuberculosis, as indicated by the size of lung lesions on chest X-ray films. Furthermore, chemotherapy resulted in a significant fall in plasma OPN levels. In patients with tuberculosis, plasma OPN concentrations correlated significantly with those of interleukin (IL)-12. In vitro experiments showed that OPN production by peripheral blood mononuclear cells infected with Mycobacterium bovis bacillus Calmette-Guerin preceded the synthesis of IL-12 and interferon-gamma and that the neutralizing anti-OPN monoclonal antibody significantly reduced the production of IL-12 and interferon-gamma. Our results suggest that OPN may be involved in the pathologic process associated with active pulmonary tuberculosis by inducing IL-12-mediated type 1 T helper cell responses.

8187. Long R, Gardam M. Tumour necrosis factor-alpha inhibitors and the reactivation of latent tuberculosis infection. CMAJ. 2003 Apr 29;168(9):1153-6. Review. No abstract.

8188. Marei AM, El-Behedy EM, Mohtady HA, Afify AF. Evaluation of a rapid bacteriophage-based method for the detection of Mycobacterium tuberculosis in clinical samples. J Med Microbiol. 2003 Apr;52(Pt 4):331-5.

Rapid, sensitive and low-cost methods are needed urgently for the detection of Mycobacterium tuberculosis in clinical samples, especially in developing countries. To this end, the clinical performance of FASTPlaqueTB(TM) (a bacteriophage-based method) has been studied in parallel with microscopy, standard microbiological culture and in-house IS6110-based PCR methods. A total of 64 samples, including 42 sputum samples and 22 urine samples, were tested in this study. The sensitivity, specificity and overall accuracy values for the FASTPlaqueTB assay relative to that of culture were respectively 76.5, 95 and 90 %. The corresponding values for the in-house IS6110-based PCR assay were 88, 91 and 90 % and, for Ziehl-Neelsen staining, were 59, 95 and 85 %. FASTPlaqueTB gave better clinical performance with urine samples than with sputum samples (sensitivity, specificity and overall accuracy were 100 % with urine samples and 64, 93 and 84 % with sputum samples). The 100 % sensitivity of FASTPlaqueTB was higher than that of the corresponding values for PCR (67 %) with urine samples. In conclusion, FASTPlaqueTB proved to be sensitive, cheap relative to the PCR and rapid. It is able to detect M. tuberculosis in clinical samples within 1 day, reducing the time to diagnosis in comparison with culture.

8189. Mazurek GH, LoBue PA, Daley CL, Bernardo J, Lardizabal AA, Iademarco MF. Detection of Mycobacterium tuberculosis infection by whole-blood interferon-gamma release assay. Clin Infect Dis. 2003 May 1;36(9):1207-8; author reply 1209-10. No abstract.

8190. Ojha UK, Study of serum magnesium estimation in pulmonary tuberculosis of different severity. Indian med J 2002, 96(9), 245-6. ISA 005711, Vol 39, No 6, 16 Mar 2003.

8191. Park JS, Kim YS, Jee YK, Myong NH, Lee KY. Interleukin-8 production in tuberculous pleurisy: role of mesothelial cells stimulated by cytokine network involving tumour necrosis factor-alpha and interleukin-1 beta. Scand J Immunol. 2003 May;57(5):463-9.

Interleukin-8 (IL-8) plays an important role in the host immune response to Mycobacterium tuberculosis by recruiting inflammatory cells to the site of infection. Here, we investigated the role of pleural macrophages and mesothelial cells in the production of IL-8 in tuberculous pleurisy. Large concentrations of IL-8 were detected in tuberculous pleural effusions, but not in pleural effusions associated with congestive heart failure (CHF). Tuberculous pleural macrophages and M. tuberculosis-infected CHF pleural macrophages produced large concentrations of IL-8. When immunohistochemistry was performed on pleural tissues, antigenic IL-8 was detected in the mesothelial cells lining the tuberculous pleura. Direct stimulation of cultured CHF pleural mesothelial cells with M. tuberculosis induced IL-8 secretion. However, conditioned media from M. tuberculosis-infected pleural macrophages (CoMTB) induced greater mesothelial cell IL-8 secretion. Tumour necrosis factor-alpha (TNF-alpha) and IL-1beta induced mesothelial cell IL-8 mRNA expression, and neutralizing anti-TNF-alpha antibody and IL-1 receptor antagonist nearly completely obliterated CoMTB-induced mesothelial cell IL-8 mRNA expression and protein secretion. These findings demonstrate that both pleural macrophages and mesothelial cells produce IL-8 in tuberculous pleurisy, and cytokines produced by M. tuberculosis-infected macrophages mediate mesothelial cell IL-8 production.

8192. Radford AJ, Rothel JS. Diagnosis of tuberculosis. Lancet. 2003 Jun 14;361(9374):2083. No abstract.

8193. Rodger A, Jaffar S, Paynter S, Hayward A, Carless J, Maguire H. Delay in the diagnosis of pulmonary tuberculosis, London, 1998-2000: analysis of surveillance data. BMJ. 2003 Apr 26;326(7395):909-10. No abstract.

8194. Rothel JS, Radford AJ. Comparison of tuberculosis tests: finding truth or confirming prejudice? Clin Infect Dis. 2003 May 1;36(9):1206-7; author reply 1209-10. No abstract.

8195. Shenai S, Rodrigues C, Mehta AP. Evaluation of a new phage amplification technology for rapid diagnosis of tuberculosis. Indian J med Microbiol 2002, 20(4), 194-9. ISA 000730, Vol 39, No 1, 1 Jan 2003

8196. Siddiqi K, Lambert ML, Walley J. Clinical diagnosis of smear-negative pulmonary tuberculosis in low-income countries: the current evidence. Lancet Infect Dis. 2003 May;3(5):288-96. Review.

Sputum smear examination for acid-fast bacilli (AFB) can diagnose up to 50-60% of cases of pulmonary tuberculosis in well-equipped laboratories. In low-income countries, poor access to high-quality microscopy services contributes to even lower rates of AFB detection. Furthermore, in countries with high prevalence of both pulmonary tuberculosis and HIV infection, the detection rate is even lower owing to the paucibacillary nature of pulmonary tuberculosis in patients with HIV infection. In the absence of positive sputum smears for AFB, at primary care level, most cases of pulmonary tuberculosis are diagnosed on the basis of clinical and radiological indicators. This review aims to evaluate various criteria, algorithms, scoring systems, and clinical indicators used in low-income countries in the diagnosis of pulmonary tuberculosis in people with suspected tuberculosis but repeated negative sputum smears. Several algorithms and clinical scoring systems based on local epidemiology have been developed to predict smear-negative tuberculosis. Few of these have been validated within the local context. However, in areas where smear-negative tuberculosis poses a major public-health problem, these algorithms may be useful to national tuberculosis programmes by providing a starting point for development their own context-specific diagnostic guidelines.

8197. Usha K, Saroja S. Nutritional and Biochemical Profiles in Children afflicated with pulmonary tuberculosis. Indian J Nutr Diet 2002, 39(10), 439-43. ISA 004785, Vol 39, No 5, 1 Mar 2003.

8198. Wood PR, Jones SL. Diagnosis of tuberculosis. Lancet. 2003 Jun 14;361(9374):2081-2; author reply 2082-3. No abstract.

Pathogenesis:

8199. Adams CH, Werely CJ, Victor TC, Hoal EG, Rossouw G, van Helden PD. Allele frequencies for glutathione S-transferase and N-cetyltransferase 2 differ in African population groups and may be associated with oesophageal cancer or tuberculosis incidence. Clin Chem Lab Med. 2003 Apr;41(4):600-5.

Glutathione S-transferase (GST) and arylamine N-acetyltransferase 2 (NAT2) metabolise many environmental and chemotherapeutic agents, which influence susceptibility to disease. Polymorphisms in these enzymes result in different host phenotypes and contribute to different disease profiles or responses to toxic or chemotherapeutic agents, depending on their frequency in different populations. GST and NAT2 polymorphisms were investigated in different population groups, including African populations, and a range of allelic frequencies have been observed. The GSTM1 null genotype frequency, reported in this paper in two South African ethnic groups, is the lowest reported (0.19-0.21). In contrast, these same groups have a high GSTT1 null frequency (0.41-0.54), which is considerably higher than in African-Americans, or other Africans. The GSTT1 null frequency is comparable to the Chinese, a population with a very high oesophageal cancer incidence, similar to that in the African group. The frequency of the GSTPi Val105 variant in the South African Xhosas was also high (0.53), differing significantly from the low frequency in other Africans. These variants could therefore be associated with high cancer susceptibility. In addition, the high proportion of NAT2 "fast" alleles may partially explain the high tuberculosis prevalence in South Africans, due to reduced isoniazid efficacy in the presence of rapid acetylation.

8200. Al-Attiyah R, Shaban FA, Wiker HG, Oftung F, Mustafa AS. Synthetic peptides identify promiscuous human Th1 cell epitopes of the secreted mycobacterial antigen MPB70. Infect Immun. 2003 Apr;71(4):1953-60.

MPB70 is a secreted protein of Mycobacterium bovis and Mycobacterium tuberculosis which stimulates both cellular and humoral immune responses during infection with bovine and human tubercle bacilli. In addition, vaccination with MPB70 has been shown to induce Th1 cell responses and protection in animal models of tuberculosis. The present study was carried out to map the dominant human Th1 cell epitopes of MPB70 in relation to major histocompatibility complex (MHC) class II restriction in healthy subjects showing strong T-cell responses to complex mycobacterial antigens. Peripheral blood mononuclear cells (PBMC) from HLA-DR-typed donors were tested with complex mycobacterial antigens (whole-cell M. tuberculosis and M. tuberculosis culture filtrates), with MPB70 purified from the culture filtrate of M. bovis BCG Tokyo, and with 13 synthetic peptides (25-mers overlapping by 10 residues) covering the sequence of MPB70. The donors that responded to the complex antigens and MPB70 also responded to the cocktail of synthetic MPB70 peptides. Testing of PBMC with individual peptides showed that peptides p5 (amino acids [aa] 61 to 85), p6 (aa 76 to 100), p8 (aa 106 to 130), p12 (aa 166 to 190), and p13 (aa 181 to 193) were most frequently recognized in proliferation and gamma interferon (IFN-gamma) assays. Testing of antigen-specific CD4(+) T-cell lines with the individual peptides of MPB70 confirmed that peptides p8, p12, and p13 contain immunodominant Th1 cell epitopes of MPB70. MHC restriction analysis with HLA-typed donors showed that MPB70 and its immunodominant peptides were presented to T cells promiscuously. The T-cell lines responding to MPB70 and peptides p8, p12, and p13 in IFN-gamma assays mediated antigen-peptide-specific cytotoxic activity against monocytes/macrophages pulsed with the whole-protein antigen or the peptides. In conclusion, the promiscuous recognition of MPB70 and its immunodominant peptide defined epitopes (aa 106 to 130 and 166 to 193) by IFN-gamma-producing Th1 cells supports possible application of this secreted antigen to subunit vaccine design.

8201. Corbett EL, Watt CJ, Walker N, Maher D, Williams BG, Raviglione MC, Dye C. The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. Arch Intern Med. 2003 May 12;163(9):1009-21. Review.

BACKGROUND: The increasing global burden of tuberculosis (TB) is linked to human immunodeficiency virus (HIV) infection. METHODS: We reviewed data from notifications of TB cases, cohort treatment outcomes, surveys of Mycobacterium tuberculosis infection, and HIV prevalence in patients with TB and other subgroups. Information was collated from published literature and databases held by the World Health Organization (WHO), the Joint United Nations Programme on HIV/Acquired Immunodeficiency Syndrome (UNAIDS), the US Census Bureau, and the US Centers for Disease Control and Prevention. RESULTS: There were an estimated 8.3 million (5th-95th centiles, 7.3-9.2 million) new TB cases in 2000 (137/100,000 population; range, 121/100,000-151/100,000). Tuberculosis incidence rates were highest in the WHO African Region (290/100,000 per year; range, 265/100,000-331/100,000), as was the annual rate of increase in the number of cases (6%). Nine percent (7%-12%) of all new TB cases in adults (aged 15-49 years) were attributable to HIV infection, but the proportion was much greater in the WHO African Region (31%) and some industrialized countries, notably the United States (26%). There were an estimated 1.8 million (5th-95th centiles, 1.6-2.2 million) deaths from TB, of which 12% (226 000) were attributable to HIV. Tuberculosis was the cause of 11% of all adult AIDS deaths. The prevalence of M tuberculosis-HIV coinfection in adults was 0.36% (11 million people). Coinfection prevalence rates equaled or exceeded 5% in 8 African countries. In South Africa alone there were 2 million coinfected adults. CONCLUSIONS: The HIV pandemic presents a massive challenge to global TB control. The prevention of HIV and TB, the extension of WHO DOTS programs, and a focused effort to control HIV-related TB in areas of high HIV prevalence are matters of great urgency.

8202. Dale JW, Al-Ghusein H, Al-Hashmi S, Butcher P, Dickens AL, Drobniewski F, Forbes KJ, Gillespie SH, Lamprecht D, McHugh TD, Pitman R, Rastogi N, Smith AT, Sola C, Yesilkaya H. Evolutionary relationships among strains of Mycobacterium tuberculosis with few copies of IS6110. J Bacteriol. 2003 Apr;185(8):2555-62.

Molecular typing of Mycobacterium tuberculosis by using IS6110 shows low discrimination when there are fewer than five copies of the insertion sequence. Using a collection of such isolates from a study of the epidemiology of tuberculosis in London, we have shown a substantial degree of congruence between IS6110 patterns and both spoligotype and PGRS type. This indicates that the IS6110 types mainly represent distinct families of strains rather than arising through the convergent insertion of IS6110 into favored positions. This is supported by identification of the genomic sites of the insertion of IS6110 in these strains. The combined data enable identification of the putative evolutionary relationships of these strains, comprising three lineages broadly associated with patients born in South Asia (India and Pakistan), Africa, and Europe, respectively. These lineages appear to be quite distinct from M. tuberculosis isolates with multiple copies of IS6110.

8203. de Oliveira MM, da Silva Rocha A, Cardoso Oelemann M, Gomes HM, Fonseca L, Werneck-Barreto AM, Valim AM, Rossetti ML, Rossau R, Mijs W, Vanderborght B, Suffys P. Rapid detection of resistance against rifampicin in isolates of Mycobacterium tuberculosis from Brazilian patients using a reverse-phase hybridization assay. J Microbiol Methods. 2003 Jun;53(3):335-42.

The main objective of this study was to evaluate INNO-LiPA Rif.TB and to determine the frequency of mutations in rpoB in rifampicin-resistant Mycobacterium tuberculosis isolates of Brazilian tuberculosis patients. We used the reverse hybridization assay on 113 resistant and 15 sensitive clinical isolates of M. tuberculosis and on reference strains belonging to 37 different species. All MTB complex strains and none of the other strains reacted with the MTB complex-specific probe, meaning that the assay is 100% specific and 100% sensitive for detection of strains of the MTB complex. In 80 resistant strains, mutations causing S531L (n=55), H526Y (n=9), H526D (n=12) or D516V (n=9) were detected while in 30 strains, mutations were present but their exact nature was not determined by the assay (DeltaS patterns). All sensitive strains had the sensitive genotype while among resistant isolates, a sensitive genotype was obtained in three due to the absence of mutations in the hot spot region, demonstrating an assay accuracy of 97.6% for detection of drug susceptibility. In 10 resistant cultures, two or more mutations were detected and in five, mixed sensitive and resistant genotypes were observed. The sensitivity of the assay for detection of resistant organisms in a mixture with sensitive ones were 2% and 70%, respectively, considering the appearance and disappearance of the R2 and S2 bands. The sensitivity to detect heteroresistance is similar to that of the proportion method when a specific probe for the mutation is present but the performance of the assay in the patient population will depend on the frequency of mutation distribution.

8204. Hovell M, Blumberg E, Gil-Trejo L, Vera A, Kelley N, Sipan C, Hofstetter CR, Marshall S, Berg J, Friedman L, Catanzaro A, Moser K. Predictors of adherence to treatment for latent tuberculosis infection in high-risk Latino adolescents: a behavioral epidemiological analysis. Soc Sci Med. 2003 Apr;56(8):1789-96.

The objective was to test whether theoretical variables predict adherence to treatment for latent tuberculosis infection in high-risk Latino adolescents. 286 Latino adolescents, age 13-18 years, were recruited from 10 middle/high schools in San Diego County, San Diego, USA. Participants completed a baseline interview and up to 9 monthly interviews. The cumulative number of pills consumed in 9 months was regressed on 16 independent variables, entered hierarchically in seven blocks. The final model accounted for 25% of the variance in adherence to isoniazid (INH), F (16, 230)=4.69, p<0.001. Adherence counseling (+), age (-), grades (+), being bicultural (+), and risk behaviors (-) were significantly related to adherence. Learning theories presume that adherence to medical regimens requires social support and freedom from physical and social barriers. Results support these theories. Future studies should explore additional precepts in order to identify additional predictors and to maximize adherence to INH among Latino adolescents and other high-risk populations. Doing so should decrease the risk of active TB among high-risk racial/ethnic and foreign-born populations.

8205. Mazzarelli G, Rindi L, Piccoli P, Scarparo C, Garzelli C, Tortoli E. Evaluation of the BDProbeTec ET system for direct detection of Mycobacterium tuberculosis in pulmonary and extrapulmonary samples: a multicenter study. J Clin Microbiol. 2003 Apr;41(4):1779-82.

We evaluated the BDProbeTec ET system (Becton Dickinson, Sparks, Md.), a strand displacement amplification-based technique, for direct detection of Mycobacterium tuberculosis in 867 clinical samples. Of 294 extrapulmonary specimens, 52 had positive results by both BDProbeTec ET and culture and 209 had negative results by both methods; sensitivity and specificity were 76.5 and 95.9%, respectively. After resolution of discrepancies, the sensitivity rose to 77.8%.

8206. Venketaraman V, Dayaram YK, Amin AG, Ngo R, Green RM, Talaue MT, Mann J, Connell ND. Role of glutathione in macrophage control of mycobacteria. Infect Immun. 2003 Apr;71(4):1864-71.

Reactive oxygen and nitrogen intermediates are important antimicrobial defense mechanisms of macrophages and other phagocytic cells. While reactive nitrogen intermediates have been shown to play an important role in tuberculosis control in the murine system, their role in human disease is not clearly established. Glutathione, a tripeptide and antioxidant, is synthesized at high levels by cells during reactive oxygen intermediate and nitrogen intermediate production. Glutathione has been recently shown to play an important role in apoptosis and to regulate antigen-presenting-cell functions. Glutathione also serves as a carrier molecule for nitric oxide, in the form of S-nitrosoglutathione. Previous work from this laboratory has shown that glutathione and S-nitrosoglutathione are directly toxic to mycobacteria. A mutant strain of Mycobacterium bovis BCG, defective in the transport of small peptides such as glutathione, is resistant to the toxic effect of glutathione and S-nitrosoglutathione. Using the peptide transport mutant as a tool, we investigated the role of glutathione and S-nitrosoglutathione in animal and human macrophages in controlling intracellular mycobacterial growth.

Vaccines:

8207. Young DB. Building a better tuberculosis vaccine. Nat Med. 2003 May;9(5):503-4. No abstract.

Therapy:

8208. Almeida D, Rodrigues C, Udwadia ZF, Lalvani A, Gothi GD, Mehta P, Mehta A. Incidence of multidrug-resistant tuberculosis in urban and rural India and implications for prevention. Clin Infect Dis. 2003 Jun 15;36(12):e152-4. Epub 2003 Jun 03.

We compared the incidence of multidrug resistance in 150 consecutive Mycobacterium tuberculosis isolates obtained from a rural center (in Sakawar, India) and an urban tertiary care center (in Mumbai, India). The study highlights an alarmingly high percentage of multidrug-resistant M. tuberculosis isolates in Mumbai (51%) as compared with that at the rural center (2%).

8209. Praveen Kumar; Niraj Sharma. Primary MDR TB of the breast. Indian Journal of Chest Diseases and Allied Sciences. 2003 Jan-Mar; 45(1): 63-5

ABSTRACT: A-28-year-old, lactating lady presented to us with left-sided breast abscess and lymph node enlargement in the left axillary region for the past one and a half months. Investigations revealed the breast abscess and axillary lymphadenopathy were tubercular in origin. The patient was put on standard four-drug anti-tubercular treatment (rifampicin, isoniazid, ethambutol and pyrazinamide). The patients did not respond to the intensive four-drug therapy, which was continued for three months. The culture isolate of the breast abscess grew M. tuberculosis, which was resistant to isoniazid, rifampicin and streptomycin. The patient was then retreated with a regimen comprising kanamycin, ofloxacin, ethionamide para-amino salicyclic acid (PAS), pyrazinamide and isoniazid, from which the patient benefited and recovered.

8210. Puri MM; Arora VK. Role of gallium arsenide laser irradiation at 890 nm as an adjunctive to anti-tuberculosis drugs in the treatment of pulmonary tuberculosis. Indian Journal of Chest Diseases and Allied Sciences. 2003 Jan-Mar; 45(1): 19-23.

ABSTRACT: Background. Tuberculosis is a global emergency with about nine million people developing disease every year. The long duration of treatment has emerged as a major obstacle in the control of tuberculosis. There is a need for development of new drugs and or shortened therapy. Methods. The present study was carried out to explore whether any benefit could be achieved by the addition of low level energy laser therapy (LLLT) to the conventional anti-tubercular chemotherapy. One-hundred-thirty new sputum smear positive patients of pulmonary tuberculosis were enrolled to evaluate the bio-stimulatory effects of Gallium Arsenide laser irradiation at 890 nm, as an adjuvant therapy. These patients were randomly divided into two groups to received either LLLT or sham irradiation (Control) concomitantly with anti-tuberculosis chemotherapy. Results. The patients treated with semiconductor laser as an adjuvant therapy along with antituberculosis drugs had a faster clearance of tubercle bacilli from the sputum as compared to the control group (P value at:45 days=0.1392, 60days=0.0117,75days=0.00805,90days=0.00739). Conclusions. These findings provide preliminary evidence that low level laser therapy with Gallium Arsenide laser may be a promising adjunctive therapy for patients with tuberculosis. Faster conversion of sputum should prevent the development of resistant mutants.

8211. Sharma DC. India makes poor tuberculosis progress in 2002. Lancet Infect Dis. 2003 May;3(5):265. No abstract.

8212. Zittermann A. Vitamin D in preventive medicine: are we ignoring the evidence? Br J Nutr. 2003 May;89(5):552-72. Review.

Vitamin D is metabolised by a hepatic 25-hydroxylase into 25-hydroxyvitamin D (25(OH)D) and by a renal 1alpha-hydroxylase into the vitamin D hormone calcitriol. Calcitriol receptors are present in more than thirty different tissues. Apart from the kidney, several tissues also possess the enzyme 1alpha-hydroxylase, which is able to use circulating 25(OH)D as a substrate. Serum levels of 25(OH)D are the best indicator to assess vitamin D deficiency, insufficiency, hypovitaminosis, adequacy, and toxicity. European children and young adults often have circulating 25(OH)D levels in the insufficiency range during wintertime. Elderly subjects have mean 25(OH)D levels in the insufficiency range throughout the year. In institutionalized subjects 25(OH)D levels are often in the deficiency range. There is now general agreement that a low vitamin D status is involved in the pathogenesis of osteoporosis. Moreover, vitamin D insufficiency can lead to a disturbed muscle function. Epidemiological data also indicate a low vitamin D status in tuberculosis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases, hypertension, and specific types of cancer. Some intervention trials have demonstrated that supplementation with vitamin D or its metabolites is able: (i) to reduce blood pressure in hypertensive patients; (ii) to improve blood glucose levels in diabetics; (iii) to improve symptoms of rheumatoid arthritis and multiple sclerosis. The oral dose necessary to achieve adequate serum 25(OH)D levels is probably much higher than the current recommendations of 5-15 microg/d.

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