Res Microbiol, 2000 Mar, 151(2), 135 - 42 Comparative genomics of the leprosy and tubercle bacilli; Brosch R et al.; To achieve the quantum leap in understanding required to overcome two major human diseases, leprosy and tuberculosis, systematic and comparative genome analysis has been undertaken . New insight into the biology of their causative agents has been obtained and the principle findings are reported here. Int J Tuberc Lung Dis, 2000 Jun, 4(6), 528 - 36 Quantitative sputum bacillary load during rifampin-containing short course chemotherapy in human immunodeficiency virus-infected and non-infected adults with pulmonary tuberculosis; Joloba ML et al.; SETTING: National Tuberculosis (TB) Treatment Centre, Mulago Hospital and Joint Clinical Research Centre, Kampala, Uganda . OBJECTIVE: To compare the quantitative sputum bacillary load between TB patients infected with the human immunodeficiency virus (HIV) and those non-infected, during treatment with standard short course chemotherapy (SCC) . DESIGN: To compare clinical characteristics and quantitative sputum bacillary load as measured by quantitative acid-fast bacilli (AFB) smears, colony forming unit (cfu) assay and time until positive culture in the BACTEC radiometric liquid system between 14 HIV-infected and 22 non-HIV-infected adults with initial episodes of smear-positive pulmonary TB at baseline and during treatment with standard four-drug SCC . RESULTS: Other than cavitation (P = 0.042) and adenopathy (P = 0.03), which were more common among non-HIV-infected and HIV-infected patients, respectively, there were no significant differences in baseline demographic, clinical, radiological and laboratory characteristics between the groups . Mean pretreatment sputum bacillary burden (6.5+/-0.51 log10 AFB/ml, 5.91+/-0.91 log10 cfu/ml and 1.8+/-1.7 days until positive BACTEC culture for HIV-infected patients and 6.32+/-0.85 log10 AFB/ml, 5.58+/-0.68 log10 cfu/ml and 1+/-1.2 days until positive BACTC culture for non-HIV-infected patients) were comparable between HIV-infected and non-HIV-infected patients . Clinical and bacteriological responses to standard SCC and treatment outcome did not differ between the groups . CONCLUSION: Quantitative sputum bacillary load at baseline and during SCC did not differ significantly between HIV-infected and non-HIV-infected adults with initial episodes of smear-positive TB. Int J Tuberc Lung Dis, 2000 Jun, 4(6), 513 - 8 Trial-of-antibiotic algorithm for the diagnosis of tuberculosis in a district hospital in a developing country with high HIV prevalence; Wilkinson D et al.; OBJECTIVE: To evaluate a diagnostic algorithm for pulmonary tuberculosis based on smear microscopy and objective response to trial of antibiotics . SETTING: Adult medical wards, Hlabisa Hospital, South Africa, 1996-1997 . METHODS: Adults with chronic chest symptoms and abnormal chest X-ray had sputum examined for Ziehl-Neelsen stained acid-fast bacilli by light microscopy . Those with negative smears were treated with amoxycillin for 5 days and assessed . Those who had not improved were treated with erythromycin for 5 days and reassessed . Response was compared with mycobacterial culture . RESULTS: Of 280 suspects who completed the diagnostic pathway, 160 (57%) had a positive smear, 46 (17%) responded to amoxycillin, 34 (12%) responded to erythromycin and 40 (14%) were treated as smear-negative tuberculosis . The sensitivity (89%) and specificity (84%) of the full algorithm for culture-positive tuberculosis were high . However, 11 patients (positive predictive value {PPV} 95%) were incorrectly diagnosed with tuberculosis, and 24 cases of tuberculosis (negative predictive value {NPV} 70%) were not identified . NPV improved to 75% when anaemia was included as a predictor . Algorithm performance was independent of human immunodeficiency virus status . CONCLUSION: Sputum smear microscopy plus trial of antibiotic algorithm among a selected group of tuberculosis suspects may increase diagnostic accuracy in district hospitals in developing countries. Indian J Pathol Microbiol, 1999 Jul, 42(3), 339 - 43 Association of tuberculosis with malignancy at KIMIO--an oncology centre; Kumar RR et al.; The association of tuberculosis and malignancy was studied at an oncology centre in Bangalore . The study period was from January 1981 to December 1995 . A total of 8779 clinical material obtained from patients were screened for Mycobacterium tuberculosis infection . Out of which 675 were positive for acidfast bacilli, 385 from non malignant conditions and 290 from malignant conditions . Highest incidence is seen in Head and Neck cancer (42%) followed by Gastrointestinal cancer (14.1%), Lung cancer (13.8%), Haematological cancer (10.7%), Reproductive cancer (10.3%) and miscellaneous group (9%), Antibiogram of Mycobacterial cultures was done in 282 subjects . Resistance patterns to antitubercular drugs showed highest with Isonicotinic acid hydrochloride (INH) (17.7%), followed by para amino salicylic acid (PAS) (8.5%), Streptomycin (SM) (6.7%), Rifampicin (RIF) (4.6%) and Ethambutol (EM) (0.35%). Biotechnol Bioeng, 2000 Aug 5, 69(3), 301 - 11 Oxygen-transfer strategy and its regulation effects in serine alkaline protease production by Bacillus licheniformis; Calik P et al.; The effects of oxygen transfer on the production and product distribution in serine alkaline protease (SAP) fermentation by Bacillus licheniformis and oxygen-transfer strategy in relation to the physiology of the bacilli were investigated on a defined medium with citric acid as sole carbon source in 3.5-dm(3) batch bioreactor systems . By forming a 3 x 3 matrix with the parameters air-inlet rates of Q(O)/V(R) = 0.2, 0.5, 1.0 vvm, and agitation rates of N = 150, 500, 750 min(-1), the effects of oxygen transfer were investigated at nine different conditions . The concentrations of the product SAP and by-products, i.e., neutral protease, alpha-amylase, amino acids, and organic acids, and SAP activities were determined throughout the bioprocess . Among the constant air-flow and agitation-rate fermentations, Q(O)/V(R) = 0.5 vvm, N = 750 min(-1) oxygen-transfer conditions produced maximum SAP activity that was 500 U cm(-3), at t = 37 h . With the increase in Q(O)/V(R) and/or N, Damkohler number that is the oxygen-transfer limitation decreases; and the process passes from oxygen-transfer limited conditions to biochemical-reaction limited conditions . Further increase in SAP activity, A = 680 U cm(-3) was achieved by applying an oxygen-transfer strategy based on the analysis of the data obtained with the constant oxygen-transfer condition experiments, with a step increase in air-inlet rate, from Q(O)/V(R) = 0.2 to Q(O)/V(R) = 0.5 vvm at N = 750 min(-1) constant agitation rate at t = 24 h . Organic acids and amino acids that were excreted to the fermentation medium varied depending on the oxygen-transfer conditions . With the increase in oxygen-transfer rate acetic acid concentration increased; contrarily, with the decrease in the oxygen-transfer rate the TCA-cycle organic acids alpha-ketoglutaric and succinic acids, and gluconic acid were excreted to the fermentation broth; nevertheless, the application of the oxygen-transfer strategy prevented the increase in acetic acid concentration between t = 35-38 h . Under all the oxygen-transfer conditions, the amino acid having the highest concentration and the amino acid that was not excreted to the fermentation broth were lysine and asparagine, respectively; both of which belong to the aspartic acid-group amino acids . Further, this result indicates the requirement of the genetic regulation directed to the aspartic acid-group enzymes for the progress in SAP production in B . licheniformis . J Immunol, 2000 Jul 1, 165(1), 353 - 63 CD8+ CTL from lungs of Mycobacterium tuberculosis-infected mice express perforin in vivo and lyse infected macrophages; Serbina NV et al.; CD8+ T lymphocytes have been implicated in the protective immune response against human and murine tuberculosis . However, the functional role that this cell subset plays during the resolution of infection remains controversial . In this study, we demonstrate the presence of Mycobacterium tuberculosis-specific CD8+ CTL in the lungs and lung-draining lymph nodes of mice infected with M . tuberculosis via the aerosol or i.v . route . These cells expressed perforin in vivo and specifically recognized and lysed M . tuberculosis-infected macrophages in a perforin-dependent manner after a short period of in vitro restimulation . The efficiency of lysis of infected macrophages was dependent upon the time allowed for interaction between macrophage and M . tuberculosis bacilli . Recognition of infected targets by CD8+ CTL was beta 2-microglobulin and MHC class I dependent and was not CD1d restricted . The presented data indicate that CD8+ T cells contribute to the protective immune response during M . tuberculosis infection by exerting cytotoxic function and lysing infected macrophages. Chest, 2000 Jun, 117(6), 1685 - 9 A 10-year experience with bacteriology of acute thoracic empyema: emphasis on Klebsiella pneumoniae in patients with diabetes mellitus; Chen KY et al.; STUDY OBJECTIVES: To provide an updated evaluation of the bacteriology of acute thoracic empyema for more efficacious treatment . DESIGN:: The medical and microbiological records of all patients who received a diagnosis of acute thoracic empyema were reviewed . Based on the bacteria isolated from the pleural fluid, the patients were classified into the following four groups: aerobic or facultative Gram-positive; aerobic Gram-negative; anaerobic; and mixed . SETTING: A university-affiliated tertiary medical center . PATIENTS AND METHODS: From January 1989 to December 1998, 171 patients with a diagnosis of acute thoracic empyema were treated . A comparative analysis of the isolates from pleural effusions, the mean length of hospital stay, the mean duration of chest tube drainage, the mean duration between the onset of symptoms and the establishment of diagnosis, treatment efficacy, and the need for subsequent intervention was performed . RESULTS: A total of 163 microorganisms were isolated from the pleural fluid of 139 patients . These patients were classified according to the following types of isolates: aerobic or facultative Gram-positive (n = 47); aerobic Gram-negative (n = 59); anaerobic (n = 14); and mixed (n = 19) . Klebsiella pneumoniae was the most commonly isolated pathogen (24 . 4%) and was strongly associated with a diagnosis of diabetes mellitus . The mortality rate of patients with aerobic Gram-negative bacilli isolated was the highest (22.0%), followed by those with mixed pathogens isolated (15.7%), aerobic or facultative Gram-positive (6.4%), and anaerobic (0%) . CONCLUSIONS: The increasing incidence of acute thoracic empyema caused by Gram-negative bacilli, especially by K pneumoniae, has become an increasing problem . The isolation of aerobic Gram-negative bacilli or multiple pathogens from pleural fluid is associated with a poor prognosis and indicates a need for more aggressive antimicrobial chemotherapy. Kekkaku, 2000 May, 75(5), 389 - 94 {An outbreak of pulmonary tuberculosis in the dormitory of construction labors suspected to have been due to exogenous reinfection}; Kurasawa T et al.; We report an outbreak of pulmonary tuberculosis (TB) in a dormitory of construction labors, and this outbreak is suspected to have been caused by exogenous reinfection, based on the restriction fragment length polymorphism (RFLP) analysis and other findings . After a patient entered our hospital with active TB, 12 new other patients were discovered by contacts examination . These patients lived together in the same dormitory . They were all male and single, and were aged from 43 to 63 years old . Except one patient (No . 3) previously treated for TB for three months about 2 years ago and was suspected to be the index case of this outbreak, 12 other patients did not have a medical history of TB . The bacilli cultured from 11 patients (No . 1-11) were tested by RFLP analysis, three patterns were identified, and the fingerprints from 9 patients (No . 1-9) were identical, and the patterns of incomplete resistance of some antituberculous drugs were quite similar between No . 1-9 and No . 12 and between No . 10 and No . 13, respectively . The locations of the main lesions of TB on chest X-ray pictures were the apico-posterior segments of bilateral upper lobes . No signs suspected to indicate primary tuberculosis were detected . Considering the rate of tuberculous infection in Japan among the middle age and above as well as the identical RFLP results, most of patients in this outbreak except the index case No . 3 were suspected to have TB due to the exogenous reinfection. Diagn Mol Pathol, 2000 Jun, 9(2), 67 - 74 Molecular detection of Mycobacterium tuberculosis in tissues showing granulomatous inflammation without demonstrable acid-fast bacilli; Li JY et al.; Early diagnosis of tuberculosis (TB) is important for early medical intervention and prevention of spread of the bacteria . It is not uncommon to observe granulomatous inflammation but without demonstrable acid-fast bacilli (AFB) on Ziehl-Neelsen (ZN) staining in tissues sent for histologic examination, and the definitive diagnosis of TB cannot be made because no concurrent tissue is sent for TB culture . In this study, the authors explored the feasibility of using polymerase chain reaction (PCR) for early detection of Mycobacterium tuberculosis (Mtb) in formalin-fixed, paraffin-embedded tissues where a definite diagnosis of TB cannot be made . One hundred fifteen patients (131 paraffin blocks of biopsy specimens) with granulomatous inflammation but ZN-negative for AFB were studied . DNA was extracted from paraffin sections and amplified by PCR with the IS6110 primers (specific for the Mtb complex) and the specific 122-base pairs (bp) PCR product was detected by agarose gel electrophoresis . Sixty-eight of the 115 (59%) patients were TB-PCR positive, thus enabling definite diagnosis of TB in significant numbers of these patients in 3 days . The authors conclude that molecular diagnosis by PCR is useful for early detection of TB in histologic material where morphologic features are suggestive but not confirmatory because of negative staining for AFB. Appl Biochem Biotechnol, 2000 Spring, 84-86, 809 - 19 Anaerobic digestion from residue of industrial cassava industrialization with acidogenic and methanogenic physical separation phases; Paixao MA et al.; A trial was carried out in a continuous regimen, using a completely stirred tank reactor, at acidogenic phase, and a hybrid reactor (upflow anaerobic sludge blanket + fixed bed) at methanogenic phase at room temperature . The residue to be treated came from a flour and cassava meal industry, and the reactors operated for 300 d with affluent chemical oxygen demand (COD) concentrations of 7500, 9000, 11,000, and 14,000 mg/L . The final results showed a biogas production with a content of 80% methane and an average reduction of COD and free cyanide of nearly 96 and 98%, respectively . The separation of phases selected bacterial groups . At acidogenic phase, a predominance of propionic, n-butyric, and n-valeric acids, as well as a biomass composed of 95% fermentative bacilli, which were responsible for a 90% reduction in free cyanide concentration, was observed . At methanogenic phase, a predominance of methanogenic bacteria that came only from the Methanothrix genus was observed . The bacteria were responsible for high levels of organic matter removal and methane production. Int J Dermatol, 2000 May, 39(5), 343 - 7 Retrospective study of Mycobacterium marinum skin infections; Ang P et al.; BACKGROUND: Although infection by Mycobacterium marinum is well reported in the literature, there have been few epidemiologic studies . The purpose of this study was to review the epidemiology of patients with cutaneous M . marinum infection over a 3-year period at the National Skin Centre in Singapore . METHODS: Patients with a diagnosis of cutaneous M . marinum infection, confirmed histologically, were collated from computerized data from 1995 to 1997 . Thirty-eight patients were diagnosed as having cutaneous M . marinum infection based on history, and clinical and histologic features . RESULTS: Out of the 38 cases of M . marinum infection, there were 30 men and eight women . The age range was 14-85 years (mean: 44.7 years) . The duration of disease ranged from 1 to 132 months (mean: 19 months) . Thirteen patients (34.2%) had fish rearing as a hobby and four patients (10.5%) had occupational exposure to fish . Twelve patients (31.5%) gave a history of trauma to the disease site . All patients had biopsies of the lesions . All showed infective granulomas/granulomatous inflammation on histology . Acid-fast bacilli were identified in five out of 38 patients (13.2%) and mycobacteria were isolated in one out of 35 patients (2.9%) . Nineteen patients received treatment with cotrimoxazole-trimethoprim alone, three with minocycline alone, five with minocycline and cotrimoxazole-trimethoprim, seven with various combinations of drugs, one with excision, and three defaulted treatment . The duration of treatment ranged from 4 to 38 weeks (mean: 14.9 weeks) . Twenty-six patients (68.4%) showed clinical improvement, two (5.3%) had no response, and 10 (26.3%) were lost to follow-up . None of the patients worsened with treatment . The follow-up period ranged from 1 to 20 months (mean: 6.8 months) . CONCLUSIONS: The diagnosis of cutaneous M . marinum infection is mainly clinical, with supporting evidence from histologic features and the response to therapy . Risk factors include a history of trauma and water/fish-related hobbies or occupations . There is a poor yield of positive isolates in our experience; however, empirical treatment usually produces a good clinical response . In future, the polymerase chain reaction (PCR) technique may become more widely available as a rapid, sensitive, and specific means of diagnosis. Rev Panam Salud Publica, 2000 Apr, 7(4), 232 - 41 {Tuberculosis recurrence and its risk factors}; de Oliveira HB et al.; The persistence of tuberculosis bacilli in patients who are cured, thus causing recurrence, is an important issue . This case-control study investigated individual and institutional risk factors for relapse by analyzing independent variables related to the patient, the use of antituberculosis drugs, and the service delivered at health care institutions; 56 cases and 105 controls were interviewed . Recurrence was defined as a new tuberculosis episode after the patient had been successfully treated . Controls were selected from among patients who were treated and cured of pulmonary tuberculosis and who did not experience a relapse . Regression models were proposed to control confounding factors or effect modifiers . The variables identified as risk factors for relapse were those related to erratic patient behavior (missing medical appointments and therefore not picking up the medication, not taking the medication, taking the wrong dosage), age, and stress from life events; adverse reactions to antituberculosis drugs; and problems in the organization of health care services that resulted in patients receiving insufficient dosages or amounts of antituberculosis drugs . Receiving information regarding treatment duration provided protection against recurrence . The knowledge regarding these risk factors should result in more intensive follow-up and in more use of directly observed treatment of tuberculosis in order to prevent relapse. Nihon Kokyuki Gakkai Zasshi, 2000 Mar, 38(3), 223 - 8 {An adult case of cervico-mediastinal tuberculous lymphadenitis}; Nakamura M et al.; We reported a case of cervico-mediastinal tuberculous lymphadenitis followed by the development of pulmonary lesions and recurrent laryngeal nerve palsy . A 48-year-old man was admitted to our hospital due to fever, dry cough, and loss of body weight . He had no medical history of tuberculosis or other significant diseases . On admission, a right cervical tumor was palpable and chest X-ray films revealed widened superior mediastinum . Chest computed tomography showed multiple swollen mediastinal lymph nodes, including multiple low-density areas and contrast medium-enhanced septa and margins . Gallium-67 scintigraphy demonstrated abnormal uptake in the right cervix and mediastinum . A PPD skin test was strongly positive and ribosomal RNA of tubercle bacilli was detected in aspirated gastric juice . Although anti-tuberculous chemotherapy was initiated, fever and cough persisted, and hoarseness due to left recurrent laryngeal nerve palsy developed . One month later, chest X-ray films showed abnormal infiltration in the left upper lung field . The patient was sero-negative for HIV . To confirm the diagnosis, right supraclavicular lymph node biopsy was performed . Microscopic examination of the biopsy specimen revealed acid-fast bacilli and granuloma with central caseous necrosis surrounded by Langhans' giant cells and epithelioid cells . After 9 months of extended anti-tuberculous chemotherapy, the cervical and mediastinal masses receded and the abnormal chest X-ray shadows disappeared . Because the incidence of tuberculosis in Japan is gradually increasing among young people as well as the elderly, the differential diagnosis of this disease will become more necessary. Int J Syst Evol Microbiol, 2000 May, 50 Pt 3, 979 - 83 Bulleidia extructa gen . nov., sp . nov., isolated from the oral cavity; Downes J et al.; Five strains of anaerobic non-sporing Gram-positive bacilli isolated from advanced periodontitis (four strains) and a dentoalveolar abscess (one strain) that did not correspond to existing species were subjected to phenotypic and genetic characterization . Following 16S rDNA sequence analysis, they were found to constitute a novel branch of the low G+C Gram-positive division of the phylogenetic tree related to Erysipelothrix rhusiopathiae and Holdemania filiformis . A new genus Bulleidia, and the species Bulleidia extructa, are proposed . Growth of B . extructa in broth media was poor but was enhanced by the addition of fructose, glucose or maltose together with Tween 80 . Glucose and maltose were fermented and arginine was hydrolysed . Acetate, lactate and trace amounts of succinate were the end products of glucose fermentation . The G+C content of the DNA of the type strain is 38 mol% . The type strain of Bulleidia extructa is DSM 13220T. Bol Asoc Med P R, 1999 Jul-Dec, 91(7-12), 103 - 5 Clostridial sepsis: is death avoidable? Casiano Torres CF, Jaume Anselmi F, Ramirez Rivera J. Massive intravascular hemolysis is a rare yet often fatal complication of clostridial sepsis . The only chance for survival is an early diagnosis and prompt initiation of treatment . We report a rapidly fatal case who developed electrocardiographic changes of acute myocardial injury . Autopsy showed gas-filled bubbles and cysts in the myocardium partially filled with sporulating bacilli with the morphology of clostridia . Gas filled bubbles were also present in the lungs, liver, kidneys and spleen . The gastric mucosa showed hemorrhagic and necrotizing changes, the probable site of entry of the infection. FEMS Microbiol Rev, 2000 Jul, 24(3), 225 - 49 The roots of microbiology and the influence of Ferdinand Cohn on microbiology of the 19th century; Drews G; The beginning of modern microbiology can be traced back to the 1870s, and it was based on the development of new concepts that originated during the two preceding centuries on the role of microorganisms, new experimental methods, and discoveries in chemistry, physics, and evolutionary cell biology . The crucial progress was the isolation and growth on solid media of clone cultures arising from single cells and the demonstration that these pure cultures have specific, inheritable characteristics and metabolic capacities . The doctrine of the spontaneous generation of microorganisms, which stimulated research for a century, lost its role as an important concept . Microorganisms were discovered to be causative agents of infectious diseases and of specific metabolic processes . Microscopy techniques advanced studies on microorganisms . The discovery of sexuality and development in microorganisms and Darwin's theory of evolution contributed to the founding of microbiology as a science . Ferdinand Cohn (1828-1898), a pioneer in the developmental biology of lower plants, considerably promoted the taxonomy and physiology of bacteria, discovered the heat-resistant endospores of bacilli, and was active in applied microbiology. J Infect, 2000 Mar, 40(2), 187 - 9 Superior vena cava syndrome during the treatment of pulmonary tuberculosis in an HIV-1 infected patient; Minguez C et al.; Tuberculosis is nowadays an uncommon cause of superior vena cava syndrome (SVCS) . We report the case of an HIV-infected patient who presented with respiratory symptoms accompanied by cervical and mediastinal lymphadenopathy . Sputum examination showed acid-fast bacilli, and treatment was instituted with isoniazid, rifampicin, pyrazinamide and ethambutol . A few days later SVCS developed, presumably as a consequence of inflammatory lymphadenitis . With corticosteroids, all symptoms disappeared . To our knowledge, no cases of SVCS provoked by this kind of paradoxical reaction have been described previously. Am J Infect Control, 2000 Jun, 28(3), 244 - 50 Increasing antimicrobial resistance in gram-negative bacilli isolated from patients in intensive care units; Flournoy DJ et al.; BACKGROUND: We investigated gram-negative bacilli from patients in intensive care units to determine whether antimicrobial resistance was increasing . METHODS: Minimal inhibitory concentrations were determined by broth microdilution on 334 gram-negative bacilli collected in 1990, 1995, and 1998 . RESULTS: During the 3 study years, the types of gram-negative bacilli encountered in our intensive care units changed with proportional increases of Pseudomonas sp and decreases of inducible enterics . Dramatic increases in resistance for ceftazidime, cefotaxime, and piperacillin were paralleled between respiratory-tract isolates and inducible enterics . By 1998, ticarcillin was more active than piperacillin against most isolates except Escherichia coli and Klebsiella sp, and most isolates became more resistant to gentamicin and tobramycin . CONCLUSIONS: Continuous changes in the types of gram-negative bacilli and antimicrobial resistance complicate empirical selection of antimicrobials in the intensive care units . These complications will place more emphasis on communication and strategy formations among health care workers (nurses, physicians, laboratorians, and pharmacists) in an effort to treat infections in a timely and effective manner. J Biol Chem, 2000 Sep 8, 275(36), 28201 - 7 Identification and substrate specificity of beta -ketoacyl (acyl carrier protein) synthase III (mtFabH) from Mycobacterium tuberculosis; Choi KH et al.; The long-chain alpha-alkyl-beta-hydroxy fatty acids, termed mycolic acids, which are characteristic components of the mycobacterial cell wall are produced by successive rounds of elongation catalyzed by a multifunctional (type I) fatty acid synthase complex followed by a dissociated (type II) fatty acid synthase . In bacterial type II systems, the first initiation step in elongation is the condensation of acetyl-CoA with malonyl-acyl carrier protein (ACP) catalyzed by beta-ketoacyl-ACP III (FabH) . An open reading frame in the Mycobacterium tuberculosis genome (Rv0533c), now termed mtfabH, was 37.3% identical to Escherichia coli ecFabH and contained the Cys-His-Asn catalytic triad signature . However, the purified recombinant mtFabH clearly preferred long-chain acyl-CoA substrates rather than acyl-ACP primers and did not utilize acetyl-CoA as a primer in comparison to ecFabH . In addition, purified mtFabH was sensitive to thiolactomycin and resistant to cerulenin in an in vitro assay . However, mtFabH overexpression in Mycobacterium bovis BCG did not confer thiolactomycin resistance, suggesting that mtFabH may not be the primary target of thiolactomycin inhibition in vivo and led to several changes in the lipid composition of the bacilli . The data presented is consistent with a role for mtFabH as the pivotal link between the type I and type II fatty acid elongation systems in M . tuberculosis . This study opens up new avenues for the development of selective and novel anti-mycobacterial agents targeted against mtFabH. Microb Pathog, 2000 Jun, 28(6), 335 - 42 Identification of viable and non-viable Mycobacterium tuberculosis in mouse organs by directed RT-PCR for antigen 85B mRNA; Pai SR et al.; Mycobacterium tuberculosis (MTB) the causative organism of tuberculosis can remain dormant as a non-culturable organism, reactivate and cause disease in man and animals . There is a need for proof of viability of such organisms in order to understand the process of reactivation . PCR for bacterial DNA cannot distinguish between viable and non-viable bacilli . We have tested a previously described two tube directed reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of mRNA of antigen 85B (Ag85B) of MTB that can distinguish between viable and non-viable organisms . Using a set of external and internal primers for Ag85B, a cDNA amplified product (216 bp) was seen among simulated samples containing only viable cfus at a sensitivity of >10 and <100 cfu/ml . Eucaryotic DNA rich normal mouse lung homogenate did not interfere among these samples . The method amplified the 216 bp product also among cfu positive tissues of naturally infected mice . Finally, in a mouse model of dormancy, direct RT-PCR detected a signal among multiple tissues that were negative for cfus and hence non-culturable . Ag85B is abundantly secreted by MTB and hyper-expressed under stress conditions . Thus the method to identify its mRNA message may be useful to detect viable but dormant bacteria . Indian J Pediatr, 2000 Mar, 67(3), 197 - 9 Stethoscopes and nosocomial infection; Sengupta S et al.; Stethoscopes are an essential tool of the medical profession and can become a source of nosocomial infection . A study conducted in the Department of Pediatrics, Kasturba Medical College and Hospital showed a high carriage of methicillin resistant staphylococcus (69.76%) and multi-drug resistant Gram negative bacilli (20.93%) on regularly used stethoscopes . The antibiogram of the bacterial isolates strongly suggested these to be nosocomial strains . A verbal survey revealed that regular cleaning is not common among doctors . A policy regarding cleaning of stethoscopes with an effective disinfectant may be helpful in reducing hospital-associated infections. J Antimicrob Chemother, 2000 Jun, 45(6), 859 - 70 A multicentre study of the early bactericidal activity of anti-tuberculosis drugs; Sirgel FA et al.; The early bactericidal activities (EBAs) of 300 mg isoniazid, 18.5 mg isoniazid, 600 mg rifampicin and 800 mg ofloxacin given daily to 262 patients with newly diagnosed pulmonary tuberculosis in Cape Town, Nairobi, Madras and Hong Kong were measured by counting cfu and total acid-fast bacilli in sputum collections taken pre-treatment (S1), at 2 days (S3) and at 5 days (S6) . In Cape Town, Nairobi and Madras, the cfu findings suggested that isoniazid produced a massive kill, perhaps of actively growing organisms, during the first 2 days (mean S1-S3 EBAs of 0.636-1.006) but was almost inactive thereafter (mean S3-S6 EBAs of 0.000-0.081), whereas rifampicin maintained moderate activity against slowly growing organisms throughout the 5 days (mean S3-S6 EBAs of 0.242-0.305) . This finding suggests that EBAs measured during the 2-5 day interval might be able to assess the sterilizing activity of drugs . Ofloxacin had moderately high mean S1-S3 EBAs of 0.130-0.391 . However, in Hong Kong rifampicin appeared to be the most bactericidal drug from the start, possibly because patients had more chronic disease . A method of adjusting the cfu EBAs using total counts was devised which decreased the variability between patients within a treatment group without altering the mean cfu EBA . This resulted in a large gain in precision in Hong Kong, suggesting that their estimates were greatly affected by type II variation, due to dilution of pus by saliva and bronchial secretions, whereas small or no gains were obtained in the other three centres, suggesting that the main cause of variability was type I, due to other factors. Medicina (B Aires), 2000, 60(1), 17 - 25 {Tuberculosis focused through the molecular biology zoom}; Barrera L; The sequence of Mycobacterium tuberculosis genome set up a process of systematic research and improved the understanding of the microorganism biology, albeit the clues of its virulence and pathogenicity still remain rather cryptic . Genetic engineering did not succeed in designing more effective or convenient tools to accomplish the control of tuberculosis . Neither the bacillus variants created by mutagenesis and recombination nor the microorganism subunits (antigens, DNA) proved to be significantly better than the BCG vaccine as immunogens . Likewise, the search for novel bacterial targets and more active antibiotics has been unfruitful thus far, even though some advance in drugs formula or delivery systems is in progress . Conversely, new and ingenious instruments have been developed to optimize the epidemiological tracing and diagnosis of the disease . The finding of strain molecular markers consolidated the investigation of tuberculosis spread and revealed the expansion and prevalence of certain lineages of the bacillus in some ecological niches . Molecular methods are specific to immediately characterize the bacilli detected by microscopy or culture which resulted in rapid diagnosis build-up . This improvement is decisive for immunodepressed patients and those affected by multidrug-resistant tuberculosis . To be meaningful and precise the results produced by molecular investigations must be properly backed up by conventional microbiology, pathology and epidemiology. Int J Lepr Other Mycobact Dis, 2000 Mar, 68(1), 11 - 7 The effect of ultraviolet light radiation on Mycobacterium leprae; Truman RW et al.; Ultraviolet (UV) light is recognized as a potent sterilizing aid, but its relative effectiveness against Mycobacterium leprae has not been shown . We examined the influence of UV on the growth and metabolic activity of M . leprae harvested fresh from foot pads of nude mice . Temporary static suspensions were exposed to timed intervals of UV radiation generated from a fixed source to constitute dosages ranging from 0-12.64 x 10(4) erg/cm2 . The metabolic activity of the bacilli was indexed by the oxidation of 14C-palmitate in BACTEC 12-B vials . The long-term effects of irradiation on cell division and growth were assessed by inoculation of BALB/c mouse foot pads . The metabolic activity in BACTEC showed an immediate dose-response-related decline to a maximum of 50% of the control activity after exposure to 6.3 x 10(4) erg/cm2 . Mouse foot pad studies showed a similar dose-response pattern . Effective-dose determinations based on metabolic or foot pad data were similar . UV doses of 3.52 x 10(4) erg/cm2 resulted in an average 50% killing, and 7.73 x 10(4) erg/cm2 killed 84% of the M . leprae exposed . This UV sensitivity is similar to that reported for M . tuberculosis . UV sterilization and disinfection practices suitable for M . tuberculosis are likely to be equally effective for M . leprae. Int J Lepr Other Mycobact Dis, 2000 Mar, 68(1), 1 - 10 Inhibition of metabolism and growth of Mycobacterium leprae by gamma irradiation; Adams LB et al.; Mycobacterium leprae is uncultivable on artificial medium, but viability can be maintained without multiplication for a limited time in vitro . In this study, we evaluated gamma-irradiation (gamma-irr) as a means to kill this slowly growing organism . Freshly harvested, viable, athymic, nu/nu mouse-derived M . leprae were exposed to varying doses of gamma-irr from a 60Co source . Two indicators of bacterial viability were determined: metabolism, measured by oxidation of 14C-palmitic acid to 14CO2 in the BACTEC 460 system, and multiplication, measured by titration in the mouse foot pad . gamma-Irr of both M . leprae and M . lufu, a cultivable control mycobacterium, resulted in a dose-dependent inhibition of viability . gamma-Irr of up to 10(3) rad had little effect on the metabolic activity of either organism . For M . leprae, 10(4)-10(5) rad caused an intermediate inhibitory effect; whereas 10(6) rad yielded almost total inhibition . In the mouse foot pad assay, up to 10(4) rad had little effect on M . leprae growth; however, 10(5) rad resulted in at least a 2-log reduction in the number of bacilli recovered and no M . leprae growth was measurable after exposure to 10(6) rad . With M . lufu, 10(5) rad inhibited metabolic activity by 99% and caused > or = 2-log reduction in the number of colony forming units (CFU) . No CFU of M . lufu were recovered after exposure to 10(6) rad . Scanning electron microscopy revealed the presence of some aberrant protrusions on the cell surface of lethally irradiated M . leprae; whereas boiling and autoclaving caused obvious morphological denaturation . These data suggest that gamma-irr is an effective way to kill M . leprae without causing extensive damage to the cell architecture . Killing M . leprae by gamma-irr may be preferable when comparing cellular responses to live versus dead bacilli in vitro and in vivo. Acta Cytol, 2000 May-Jun, 44(3), 297 - 300 Acid-fast bacilli in fine needle aspiration smears from tuberculous lymph nodes . Where to look for them; Prasoon D; OBJECTIVE: To analyze where acid-fast bacilli (AFB) are most often seen in smears prepared from tuberculous lymph nodes . STUDY DESIGN: Patients referred for fine needle aspiration cytology for evaluation of lymphadenopathy between March 1994 and June 1998 were analyzed . Only those cases clinically and therapeutically proven to be tuberculous were included in the study . RESULTS: Of 783 cases analyzed, 213 (27.2%) were tuberculous . Aspirates obtained were of three types: blood-mixed particles, caseous material and pus . Five cytologic pictures were seen: epithelioid cell granulomas alone or with coexistent necrosis, AFB or both, and necrosis with AFB . AFB were most often seen in purulent aspirates, followed by caseous and least often in blood-mixed particles . Granulomas were most often seen when the aspirate was blood-mixed particles, followed by caseous and, least often, pus . CONCLUSION: AFB detection should be carried out on all suspected tuberculous patients . The relationship between the presence of granuloma and of AFB is inverse . The chance of finding AFB is highest in patients presenting with a cold abscess and yielding pus on aspiration followed by patients who yield caseous material on aspiration. Acta Cytol, 2000 May-Jun, 44(3), 292 - 6 Tuberculosis of the breast . A cytomorphologic study; Kakkar S et al.; OBJECTIVE: Extrapulmonary tuberculosis occurring in the breast is rare despite the fact that 1-2 billion people worldwide suffer from tuberculosis . The aim of this study was to examine the cytomorphology of breast tuberculosis (breast TB) and to review the literature . STUDY DESIGN: Old records from the Cytopathology Laboratory, All India Institute of Medical Sciences, were reviewed from January 1980 to December 1998 . Cases of breast TB where a cytologic diagnosis was rendered or a histologic diagnosis with prior fine needle aspiration cytology (FNAC) was available were selected . These slides were reviewed for determining the cytologic findings . RESULTS: One hundred sixty cases of breast TB were included in the study . Six males and 154 females with a clinical suspicion of carcinoma had undergone FNA that was reported as TB . The majority of the patients (111) were in the reproductive age group, 21-40 years . Of the 160 cases, 118 (73.75%) had cytomorphology diagnostic of tuberculosis--epithelioid cell granulomas with caseous necrosis . Eleven of the remaining 42 cases were positive for acid-fast bacilli (AFB) on Ziehl-Neelsen (ZN) staining, while 31 cases were confirmed to be tubercular on histology . ZN staining was done in 44 cases, and AFB were demonstrated in only 38.6% of cases . CONCLUSION: Up to 73% of breast TB can be confidently diagnosed when both epithelioid cell granulomas and necrosis are present . Also, the possibility that a woman in the reproductive age group who presents with a palpable lump in the breast may have tuberculosis must be kept in mind, especially as the incidence of breast TB may increase in the future with the global spread of AIDS. Tidsskr Nor Laegeforen, 2000 Feb 10, 120(4), 445 - 6 {Tuberculosis of the hand}; Kali L et al.; Since 1985 there has been an increase in the incidence of skeletal tuberculosis in Denmark . This increase is attributed to a high incidence of tuberculosis among HIV-positive patients, elderly people and the increasing number of immigrants from areas with endemic tuberculosis . We present a case of hand tuberculosis in a young Somali male adult, where the tubercle bacilli find their way to the third metacarpal bone of the left hand, without a history of trauma or previous pulmonary tuberculosis . There was a delay in the treatment due to misdiagnosis of just three weeks . The abscess totally vanished two months after start of treatment. Cornea, 2000 May, 19(3), 301 - 6 Clinical and microbiological profile of Bacillus keratitis; Choudhuri KK et al.; PURPOSE: To examine the clinical and microbiological profile of Bacillus keratitis . METHODS: A retrospective review was done of all medical and laboratory records of patients with infectious keratitis in an urban tertiary level eye-care center in South India between January 1991 and June 1997 . RESULTS: Nineteen eyes of 17 patients having microbiologically proven Bacillus keratitis were reviewed . The mean age of the patients was 32.64 years (range, 3-70) . The duration of symptoms ranged from 1 day to 3 months, with 11 eyes seen within a week of onset of symptoms . Trauma (five eyes), lagophthalmos (two eyes), topical corticosteroid therapy (one eye), bullous keratopathy (two eyes), previous corneal scars (two eyes), and diabetes (one eye) were identified as predisposing factors . Severe corneal features, disproportionate to the duration of symptoms, were present in most of the eyes . Gram stain of corneal scrapings showed variably stained bacilli in eight (42.1%) cases . Polymicrobial infection was present in six eyes (two fungal, four bacterial) . Of the 16 isolates tested for in vitro antibiotic susceptibility, 100% were sensitive to gentamicin, 15 (93.75%) were sensitive to ciprofloxacin and norfloxacin, 14 (87.5%) were sensitive to chloramphenicol, and 10 (62.5%) were sensitive to cefazolin . Whereas 12 (63.1%) eyes required only medical therapy, adjunctive procedures were required in seven (36.8%) eyes . The ulcers healed (mean time to healing, 37.4+/-28.6 days) in 16 eyes (lost to follow-up, three) . Visual acuity had improved after treatment in 10 (71.4%) of 14 eyes in whom vision could be recorded . CONCLUSION: Bacillus is an unusual pathogen in the clinical setting of infectious keratitis . The infection is mostly amenable to treatment with commonly used antibiotics, and the final outcome is often satisfactory. Gynecol Oncol, 2000 Jun, 77(3), 471 - 2 A patient presenting with a pelvic mass, elevated CA-125, and fever; Straughn JM et al.; BACKGROUND: Tuberculous peritonitis is a rare event which can mimic advanced stage ovarian cancer . A pelvic mass and an elevated CA-125 is suggestive of an ovarian malignancy; however, benign conditions may be discovered, especially in the premenopausal patient . CASE: A patient with a pelvic mass, ascites, and an elevated CA-125 underwent an exploratory laparotomy for presumed ovarian cancer . Final pathology revealed pelvic tuberculosis without any pulmonary involvement . Acid-fast bacilli were confirmed with polymerase chain reaction in the surgical specimen . DISCUSSION: Pelvic tuberculosis is an uncommon gynecologic condition that presents with ascites, a pelvic mass, and fever . An elevated CA-125 is not specific for ovarian malignancy . Lab Invest, 2000 May, 80(5), 663 - 9 Association of Mycobacterium leprae with human endothelial cells in vitro; Scollard DM; Endothelial cell infection by Mycobacterium leprae has long been described histologically in all types of leprosy and in some of the acute reactions occurring in this disease . Recent evidence from experimental lepromatous neuritis indicates that M . leprae colonizes endothelial cells of epineural blood vessels even in sites of minimal infection, suggesting that interaction between these cells and M . leprae may play an important role in the selective localization of this organism to peripheral nerve . To begin to study the mechanisms involved, we have examined the interaction between M . leprae and human umbilical vein endothelial cells (HUVEC) in vitro using light microscopy, scanning and transmission electron microscopy, and confocal laser scanning microscopy . When M . leprae were added to confluent monolayers of HUVEC, uptake increased slowly to a maximum at 24 hours . Maximal percentages of infected cells were similar at ratios of organisms:cell over a range of 25:1 to 100:1 . The bacilli appeared to lie within membrane-bound vacuoles at all time points . The kinetics of association of M . leprae with HUVEC are much slower than has previously been observed with macrophages, possibly due to differences in the binding of M . leprae . Compared with other pathogens that infect endothelial cells, M . leprae also appear to be ingested more slowly, and to a more limited degree . The receptors involved in M . leprae binding to endothelial cells and the impact of intracellular infection by M . leprae on these cells remain to be determined. Mt Sinai J Med, 2000 May, 67(3), 174 - 89 Inflammatory bowel disease up to 1932; Baron JH; Inflammatory bowel diseases have been a major interest of generations of Mount Sinai Hospital gastroenterologists . Although clinical descriptions of diarrhea with or without blood go back thousands of years, clear distinctions between enteritis and ulcerative colitis were possible only in the 19th century . At that time, many case reports were published of, in retrospect, classical regional enteritis . The term "ulcerative colitis" dates from 1888; the introduction of the electric sigmoidoscope soon after made it possible to make proper diagnosis of ulcerative colitis and distinguish it from infective dysentery, membranous mucous or catarrhal colitis, and nervous diarrhea . Doctors at The Mount Sinai Hospital adopted this diagnostic approach in the 1870s and 1880s, and were particularly interested in patients with tuberculosis-like ileocecal disease without tubercle bacilli . Articles were written by Weiner in 1914, Moschcowitz and Wilensky in 1923 and 1927, and Goldfarb and Suissman in 1931 . Dr . A.A . Berg, in 1925, encouraged his assistant Leon Ginzburg to conduct a study of the inflammatory granulomatous diseases of the bowel, when Ginzburg and Gordon Oppenheimer were working in Dr . Paul Klemperer's laboratory . Initial reports came in 1927 and 1928, but Ginzburg and Oppenheimer "in conjunction with Dr . Burrill B . Crohn" presented a definitive paper, "Non-specific Granulomata of the Intestine," on May 2, 1932, to the American Gastro-Enterological Association . On May 13, 1932, Dr . Crohn presented a paper on "Terminal Ileitis" to the American Medical Association; this was published later that year with the title "Regional Ileitis: A Pathologic and Chronic Entity," under the authorship of Crohn, Ginzburg and Oppenheimer. Theor Popul Biol, 2000 May, 57(3), 235 - 47 A model for tuberculosis with exogenous reinfection; Feng Z et al.; Following primary tuberculosis (TB) infection, only approximately 10% of individuals develop active T.B . Most people are assumed to mount an effective immune response to the initial infection that limits proliferation of the bacilli and leads to long-lasting partial immunity both to further infection and to reactivation of latent bacilli remaining from the original infection . Infected individuals may develop active TB as a consequence of exogenous reinfection, i.e., acquiring a new infection from another infectious individual . Our results in this paper suggest that exogenous reinfection has a drastic effect on the qualitative dynamics of TB . The incorporation of exogenous reinfection into our TB model allows the possibility of a subcritical bifurcation at the critical value of the basic reproductive number R(0)=1, and hence the existence of multiple endemic equilibria for R(0)<1 and the exogenous reinfection rate larger than a threshold . Our results suggest that reducing R(0) to be smaller than one may not be sufficient to eradicate the disease . An additional reduction in reinfection rate may be required . These results may also partially explain the recently observed resurgence of TB . J Am Vet Med Assoc, 2000 May 15, 216(10), 1589 - 91, 1569 Mycobacterial neuritis in a cat; Paulsen DB et al.; A 2-year-old spayed female domestic shorthair cat was referred for evaluation of rapidly progressive lameness of the right hind limb, which was paralyzed . Histologic examination of biopsy specimens revealed pyogranulomatous inflammation affecting the sciatic, common peroneal, and tibial nerves, and slender, beaded, acid-fast bacilli within macrophages, nerve fibers, and degenerate axons . A diagnosis of mycobacterial neuritis was made and the cat was treated with clofazimine and enrofloxacin for extended periods . Treatment was partially effective; the goal of returning normal function to the limb was not achieved, but disease progression was halted and the cat remained a viable pet. Hiroshima J Med Sci, 2000 Mar, 49(1), 83 - 92 Light and electron microscopic study of peripheral nerve damage in patients with lepromatous leprosy (LL) and borderline lepromatous leprosy (BL); Kajihara H et al.; Cutaneous branches of radial nerves in patients with lepromatous leprosy (LL) and borderline lepromatous (BL) were studied by light and electron microscopy . Foamy macrophages were found more or less in the nerve fibers of all leprosy patients and distributed in the epineurial, perineurial and endoneurial areas . In the endoneurium, the foamy macrophages were mainly located in the subperineurial and perivascular spaces . Vacuolated Schwann cells were also found in the nerve fasciculus . In electron microscopy, these foamy macrophages and vacuolated Schwann cells contained numerous small dense materials, irregular in size and shape, considered to be degenerated and fragmented mycobacterium leprae . These dense materials were found also in the cytoplasm of vascular endothelial cells . These findings suggest that mycobacteria enter into the endoneurium via the blood vessels . In our present study, on the other hand, it was very difficult to find the intact mycobacteria in the cytoplasm of the foamy macrophages, Schwann cells or endothelial cells, as well as in the Ziehl-Neelsen staining of paraffin sections . The disappearance of intact bacilli in our present study might have been caused by multi drug therapy . The myelinated nerve fibers were degenerated and disappeared in variable degrees . Degenerative changes of the myelin sheath developed from the outer layer to the inner layer with disarrangement of the lamellar structure . These findings were different from myelin destruction of peripheral nerves in Wallerian degeneration . The degenerative changes of the myelin sheath are caused by degeneration and destruction of Schwann cells in leprosy patients . Fibrosis surrounding myelinated and unmyelinated nerve fibers, i.e., periaxonal fibrosis, was found to a greater or lesser extent in the endoneurium . In the present study, it is still unclear whether the periaxonal fibrosis was due to necrosis of the Schwann cells by infection of mycobacteria or to an autoimmune mechanism such as antiperipheral nerve antibody . However, lamellated concentric fibrosis surrounding regenerative myelinated and unmyelinated nerve fibers with the disappearance of mycobacteria suggests that degenerations and regenerations of nerve axons were repeated during clinical cause . These findings indicated that autoimmune mechanisms play an important role in the pathogenesis of periaxonal fibrosis. Lepr Rev, 2000 Mar, 71(1), 34 - 42 Which physical signs help most in the diagnosis of leprosy? A proposal based on experience in the AMFES project, ALERT, Ethiopia; Saunderson P et al.; As integration of leprosy control programmes proceeds, general health staff will have responsibility for the diagnosis of most new cases of leprosy . The training required by these workers has not yet been set out in detail . In this paper the criteria for making the diagnosis of leprosy in the AMFES cohort of 594 new cases are examined . Since this study does not include details of suspects in whom leprosy was excluded on clinical grounds, true sensitivity and specificity values cannot be calculated, but the positive predictive value of the diagnostic criteria can be measured . Sensory loss in a typical skin patch is the most important sign of early leprosy, but was not present in 132 (49%) of the 268 cases with a positive skin smear . Thickening of the ulnar nerve is a valuable sign of leprosy in Ethiopia . It can be taught to health workers, who can practise by examining their own ulnar nerves . It is more likely to be present than nerve function impairment and is particularly important when skin smears are difficult to do or are unreliable . We recommend that five basic signs are used, the presence of any two being diagnostic of leprosy: Skin lesion(s) consistent with leprosy . Loss of sensation in such a lesion . Thickening of either ulnar nerve . Loss of sensation in the palm of the hand or the sole of the foot . The presence of acid-fast bacilli in skin smears . Exact policies for the diagnosis of leprosy should be worked out and validated for each national programme. Eur J Immunol, 2000 May, 30(5), 1512 - 9 Vgamma9/Vdelta2 T lymphocytes reduce the viability of intracellular Mycobacterium tuberculosis; Dieli F et al.; An effective immune response against the intracellular pathogen Mycobacterium tuberculosis is strictly dependent on T cell activation . Although this protective response mainly depends on local release of pro-inflammatory cytokines by Th1 CD4(+) T cells, contribution of Vgamma9 / Vdelta2 T lymphocytes to immune protection against this pathogen is suggested by the antimycobacterial reactivity of this subset and its ability to produce large amounts of Th1 cytokines . Here we show that Vgamma9 / Vdelta2 T lymphocytes kill macrophages harboring live M . tuberculosis . The cytotoxic activity of Vgamma9 / Vdelta2 T lymphocytes was not MHC class I or class II restricted but was blocked by anti-TCR monoclonal antibodies, thus indicating that it involved specific interaction between the TCR and the target cell . The cytotoxicity of Vgamma9 / Vdelta2 T lymphocytes was not mediated by TNF-alpha or Fas-Fas ligand, but was shown to occur through a granule-dependent mechanism that resulted in reduction of the viability of intracellular bacilli . Perforin was shown to play an important role in killing of both infected macrophages and intracellular mycobacteria . These data strongly suggest that Vgamma9 / Vdelta2 T lymphocytes contribute to the host defense against M . tuberculosis infection. Int J Tuberc Lung Dis, 2000 May, 4(5), 409 - 13 DOT or not? Direct observation of anti-tuberculosis treatment and patient outcomes, Kerala State, India; Balasubramanian VN et al.; SETTING: The Pathanamthittha District of Kerala State, India, where the directly observed treatment, short-course (DOTS) programme was started in October 1994 . OBJECTIVE: To determine the frequency with which direct observation actually occurred within a district-level DOTS programme, and the association of treatment observation with treatment outcome under programme conditions . DESIGN: This retrospective study included 200 consecutive, newly-detected, smear-positive patients registered under the project between February 1995 and February 1996 at the District Tuberculosis Centre, as well as health workers responsible for providing directly observed treatment (DOT) who were separately and confidentially interviewed . Treatment outcomes were identified from results of sputum smear examinations for acid-fast bacilli . RESULTS: Although all patients were recorded as having received DOT, more than a quarter of patients (26.5%) did not actually receive it . The 53 patients who were not directly observed were much more likely to have treatment failure or relapse, as compared to those who had received DOT (45% vs 3%, relative risk 16.6, 95% confidence intervals 6-46, P < 0.001) . Women were somewhat less likely than men (61% vs 76%, P = 0.06) to receive DOT . Non-receivers of DOT accounted for 86% (24/28) of treatment failures or relapses . CONCLUSION: Patients treated without direct observation have a substantially higher risk of adverse outcome than those treated under direct observation . To be maximally effective, the DOTS programme must be both confidential and convenient. Aust Vet J, 1999 Jun, 77(6), 392 - 7 Detection of Mycobacterium avium subsp paratuberculosis in formalin-fixed paraffin-embedded intestinal tissue by IS900 polymerase chain reaction; Whittington RJ et al.; OBJECTIVES: To evaluate and compare methods for DNA extraction from formalin-fixed, paraffin-embedded tissues and methods for detection of Mycobacterium avium subsp paratuberculosis by IS900 PCR for confirmation of Johne's disease in ruminants . DESIGN: A laboratory study . PROCEDURE: Three methods of DNA extraction of differing complexity and two PCR protocols using different pairs of IS900 primers were compared . Sensitivity and specificity were assessed using samples from ruminants with and without histological evidence of Johne's disease . RESULTS: The simplest method of DNA extraction, which involved two cycles of boiling and freezing followed by centrifugation, gave more consistent results than two methods that required solvent extraction of paraffin, proteinase digestion and DNA purification . The sensitivity of detection of M avium subsp paratuberculosis in paraffin blocks stored for 1 to 6 years from 34 cases of Johne's disease in sheep, cattle and goats was 88% for a 229 bp IS900 PCR assay and 71% for a 413 bp assay, using the detection of acid-fast bacilli by Ziehl Neelsen staining of histological sections from the same blocks as the gold standard test . PCR results correlated with the abundance of acid-fast organisms in the tissues . No false positive reactions were detected . CONCLUSION: PCR for identification of M avium subsp paratuberculosis in formalin-fixed, paraffin-embedded intestinal tissues from ruminants is a rapid and useful method . A simple method of sample preparation is effective . Amplification of short fragments of IS900 is more effective than amplification of longer fragments. Rev Inst Med Trop Sao Paulo, 2000 Mar-Apr, 42(2), 115 - 7 Cyclospora cayetanensis in sputum and stool samples; Di Gliullo AB et al.; We report the observation of acid-fast Cyclospora cayetanensis oocysts in a sputum sample . The patient, a 60 year-old, HIV negative man, was successfully treated for pulmonary tuberculosis during 1997 . On February 1998, he was admitted to our center due to loss of weight, cough with purulent expectoration, dysphonia and a radiological picture of pulmonary fibrosis . Bacilloscopic study of sputum (negative for acid-fast bacilli) stained with Ziehl-Neelsen technique showed large (8-10 microm) spherical, acid-fast Cyclospora cayetanensis oocysts . No other pathogens were isolated on cultures from this sample or from laryngeal biopsy . Serial parasitologic studies showed C . cayetanensis and also eggs of Trichuris trichiura, Ascaris lumbricoides and Hymenolepis nana and of Entamoeba coli cysts . The patient lives in the outskirts of Buenos Aires in a brick-made house with potable water and works as builder of sewers . He travelled in several occasions to the rural area of province of Tucuman which has poor sanitary conditions . C . cayetanensis is an emergent agent of diarrhea and as far as we know this is the first time the parasite is observed in respiratory samples. Pneumonol Alergol Pol, 1999, 67(9-10), 477 - 80 {Tuberculosis of the tongue}; Panek B et al.; A case of coexisting tongue and pulmonary tuberculosis is presented . A 55 year old man, farmer, an alcohol abuser, suffered from respiratory symptoms infection was admitted to Dental Surgery Department because of carcinoma of tongue suspicion . His tongue was oedematous, stiff with numerous painful ulcerations . Thin-needle biopsy of the tongue revealed granuloma typical for tuberculosis . Chest x-ray examination revealed disseminated lesions and cavernous infiltration in the lungs . Tuberculosis of lungs and tongue was recognised . Antimycobacterial treatment was introduced . 2 weeks later, tuberculous bacilli were cultured . Excellent clinical and mainly-local (tongue) improvement has been achieved. Kekkaku, 2000 Apr, 75(4), 369 - 74 {Pulmonary infection caused by Mycobacterium gordonae in immunocompetent patient}; Fujita Y et al.; We report a case of 67-year-old female immunocompetent patient admitted to our hospital because of hemoptysis . Computed tomography (CT) of the lung showed bronchiectasis in the right S5 and small nodules in the right S6 and left S5 . The cultures of sputum and bronchial washing specimen repeatedly revealed acid-fast bacilli identified as Mycobacterium gordonae (M . gordonae) by DNA-DNA hybridization (DDH) method . Thus, she was diagnosed to be infected with M . gordonae . She was treated with isoniazid, rifampicin, ethambutol and streptomycin . After treatment, the cultures of sputum and bronchial washing specimen converted to negative, and the chest CT showed improvement of small nodules . M . gordonae is a nontuberculous mycobacterium of very low pathogenic potency . Recently there have been a few reports of infection by M . gordonae not only in immunocompromised patients but also in immunocompetent patients . These cases were considered to be sensitive to initial standard antimycobacterial therapy, therefore, it is important to examine for M . gordonae in cases suspected of nontuberculous mycobacterial infection. Am J Respir Crit Care Med, 2000 May, 161(5), 1713 - 9 Diagnosis of tuberculosis by a visually detectable immunoassay for lipoarabinomannan; Chan ED et al.; Recovery of tubercle bacilli from sputum, tissue, or body fluid is the standard for the diagnosis of tuberculosis (TB) although this process is technically demanding and relatively insensitive . We have developed a simplified, visually detectable, colloidal gold-based serological assay to qualitatively detect IgG directed against the mycobacterial cell wall component lipoarabinomannan (LAM) . The objective of this investigation is to determine the accuracy of this assay in patients with active pulmonary TB and in control patients with or without latent infection . In patients with active TB, the sensitivity of anti-LAM IgG was 85 to 93% . In five patients with active TB who were smear-negative, all tested positive for anti-LAM IgG . The specificity of the test depended on the presence of tuberculous infection . In U.S . citizens comprised of young healthy adults and rheumatology patients, the specificity was 100% . In an at-risk population for tuberculous infection who were either tuberculin skin test-negative or positive, the specificity was 89% . The negative and positive predictive values of the test were 98% and 52%, respectively . We conclude that anti-LAM IgG immunoassay is relatively sensitive and specific for active TB and thus, a potentially useful screening test for active TB. Am J Respir Crit Care Med, 2000 May, 161(5), 1572 - 7 Antituberculosis activity of once-weekly rifapentine-containing regimens in mice . Long-term effectiveness with 6- and 8-month treatment regimens; Daniel N et al.; The effectiveness of various once-weekly 10 mg/kg rifapentine (P)- containing regimens for treatment of tuberculosis was assessed in mice infected intravenously with 4.3 x 10(6) colony-forming units (cfu) of Mycobacterium tuberculosis H37Rv, and treated 14 d later with various combinations of rifampin (R), P, isoniazid (H), pyrazinamide (Z), ethambutol (E), or streptomycin (S) . Control mice treated daily with either 2-mo HRZ + 4-mo HR or 2-mo HRZ + 6-mo HE were rendered spleen and lung culture-negative at 6 mo and 8 mo, respectively . Treatment failure with emergence of R-resistant bacilli occurred in all mice given once-weekly monotherapy with P for 6 mo . Once-weekly PH treatment was successful at 6 mo when it was preceded by a 2-mo daily phase with HRZ . When the initial daily phase was reduced to 2 wk, once-weekly PH-containing treatment was successful, at 6 mo, only if it was supplemented with S during the initial daily and the once-weekly phases, and at 8 mo if it was supplemented with daily H during the once-weekly phase . Without these supplements, once-weekly treatment failed in some mice with selection of R-resistant or H-resistant mutants. Am J Respir Crit Care Med, 2000 May, 161(5), 1559 - 62 A minimum 5.0 ml of sputum improves the sensitivity of acid-fast smear for Mycobacterium tuberculosis; Warren JR et al.; Detection of acid-fast bacilli (AFB) by sputum smear supports treatment decisions with pulmonary tuberculosis (TB), but smear sensitivity for Mycobacterium tuberculosis is only approximately 45 to 75% . In an effort to increase sensitivity, smears were prepared using a minimum sputum volume of 5.0 ml . Sensitivity of smears during a 39-mo period (n = 1,849) using >/= 5.0 ml of sputum was 92 . 0%, significantly greater (p < 0.001) than a sensitivity of 72.5% in a previous 24-mo period (n = 3,486) when all specimens were processed regardless of volume . All new cases of TB (n = 18) were smear-positive with >/= 5.0 ml of sputum before treatment, and all were receiving antituberculosis drugs at hospital discharge . In contrast, significantly fewer new cases of TB (14 of 26, p = 0.002) were positive before treatment when smears were prepared using sputum of any volume, and significantly fewer of these new TB cases (18 of 26, p = 0.03) were receiving treatment at hospital discharge . The eight cases without treatment were smear-negative . These results indicate that acid-fast smear using >/= 5.0 ml of sputum increases sensitivity for M . tuberculosis and accelerates treatment of TB. Cell Immunol, 2000 Apr 10, 201(1), 63 - 74 Mycobacterium tuberculosis inhibits MHC class II antigen processing in murine bone marrow macrophages; Noss EH et al.; Infection of murine bone-marrow-derived macrophages with viable Mycobacterium tuberculosis (MTB) H37Ra inhibited surface expression of MHC class II (MHC-II) molecules and processing of exogenous antigens for presentation to CD4(+) T hybridoma cells . The inhibition was not dependent on bacterial viability, since it was also produced by exposure to dead bacilli and MTB cytosol preparations, suggesting that it was initiated by a constitutively expressed bacterial component . Northern blot analysis demonstrated that MTB bacilli or cytosol decreased MHC-II mRNA, and immunoprecipitation of biosynthetically labeled molecules confirmed that MHC-II protein synthesis was diminished . Exposure to MTB or MTB cytosol also decreased expression of H2-DM, but H2-DM expression was still sufficient to catalyze conversion of MHC-II to SDS-stable dimers, a measure of MHC-II peptide loading . Thus, infection with MTB decreased both MHC-II and H2-DM expression, but diminished MHC-II synthesis provided the major limitation to antigen processing . Rev Epidemiol Sante Publique, 2000 Apr, 48(2), 127 - 36 {Transmission of tuberculosis in Gironde: epidemiologic investigation by genomic analysis of Mycobacterium tuberculosis}; Elia-Pasquet S et al.; BACKGROUND: Over the past few years, epidemiologic surveys of tuberculosis have been strengthened by new biologic technology, in particularly using RFLP (Restriction Fragment Length Polymorphism) . This technique, which identifies Mycobacterium tuberculosis patterns, has allowed to study thoroughly tuberculosis bacilli transmission and pathogenesis . First applied on tuberculosis epidemics in at risk groups, RFLP has now an interest in the epidemiologic molecular survey of urbans populations . The aim of this study is to identify, in a French department, the proportion of clustering cases of tuberculosis, suspected of recent contamination . METHODS: An active surveillance of tuberculosis allows to record systematically the cases of tuberculosis-disease in Gironde . All M . tuberculosis isolates from the patients reported in this surveillance system were processed through IS6110 based RFLP analysis . Patients were interviewed face to face before this analysis, using a standardised data collection instrument . RESULTS: 102 patients were included in 1997; the RFLP analysis of all available strains identifies a high degree of polymorphism with 71 unique patterns; twelve groups with clustering patterns were found, grouping two (nine clusters), three (two clusters) and seven patients (one cluster) each . Those cases suspected of recent transmission were younger (age<60 years) and lived in poorer conditions . Epidemiologic links were confirmed in only 35% of the 31 patients clustered . CONCLUSION: This community survey analysis has allowed to identify at risk groups for tuberculosis transmission and to strengthen tuberculosis control in Gironde. J Perinatol, 2000 Apr-May, 20(3), 151 - 6 Comparison of a closed (Trach Care MAC) with an open endotracheal suction system in small premature infants; Cordero L et al.; OBJECTIVE: To determine whether ventilated, low birth weight infants treated with closed versus open tracheal suction in a neonatal intensive care unit (NICU) differ as to airway bacterial colonization, nosocomial pneumonia, bloodstream infection (BSI), incidence and severity of bronchopulmonary dysplasia (BPD), neonatal mortality, frequency of suction, reintubation, and nurse preference . STUDY DESIGN: A total of 175 low birth weight infants (< or = 1250 gm) consecutively born (1997 to 1999), intubated, and ventilated in the delivery room were randomized on admission to the NICU to a closed (Trach Care MAC) or open suction group . Closed multi-use catheters were changed daily; open catheters were changed after every use . Two-pass endotracheal suctioning (both groups) was performed every 8 hours or as needed . Side-port connectors were not used; thus open suction required disconnection from ventilators . Tracheal aspirate cultures were obtained on admission and weekly thereafter . Nosocomial BSI (occurring after 48 hours of life) was documented by positive blood cultures . Radiographs taken before, during, and after tracheal aspirate cultures or BSIs were graded using a semiquantitative system for pneumonia and a modified score for BPD . Nurse preference regarding suction method was recorded . RESULTS: Of the original 175 patients, 10 (5 from each group) died and 32 others (16 form each group) were extubated at or before 7 days of life . The study population comprised 67 patients in the closed group and 66 in the open group who were ventilated longer than 1 week . Groups were not statistically different in terms of demographic and clinical characteristics, such as birth weight (837 vs 876 gm), ventilation (27 vs 26 days), and length of stay (49 vs 40 days) . Airway colonization with Gram-positive cocci occurred in the majority of patients by 2 weeks of life, regardless of group . A total of 39% of infants in the closed group and 44% of infants in the open group became airway colonized with Gram-negative bacilli; differences were statistically significant . No Gram-negative bacilli species was more likely to be associated with either suction . Nosocomial pneumonia was diagnosed in five patients from each group . Nosocomial BSIs occurred in six closed suction infants and five open suction infants . A comparable number of infants in each group developed severe BPD and were discharged from the hospital on oxygen . A total of 28% of closed suction patients and 27% of open suction patients died . Infants in the closed versus open group were suctioned on average 4.4 and 4.1 times per day and were reintubated 9.7 and 8.6 times per 100 ventilator days, respectively . A total of 40 of 44 NICU nurses considered closed suction to be easier to use, less time-consuming, and better tolerated by the patient . CONCLUSIONS: Closed suction obviates the physiological disadvantage of ventilator disconnection without increasing the rate of bacterial airway colonization, frequency of endotracheal suction and reintubation, duration of mechanical ventilation, length of hospitalization, incidence of nosocomial pneumonia, nosocomial BSI, severity of BPD, and neonatal mortality . Although slightly more expensive, closed suction is perceived by nursing staff to be easier, less time-consuming, and better tolerated by small premature infants requiring mechanical ventilation for > or = 1 week. Intern Med, 2000 Apr, 39(4), 309 - 12 Pulmonary mycobacteriosis caused by rifampicin-resistant Mycobacterium szulgai; Nakayama S et al.; We describe a rare case of pulmonary mycobacteriosis infected with rifampicin (RFP)-resistant Mycobacterium szulgai that was successfully eradicated with clarithromycin (CAM) treatment . An 80-year-old man was admitted to our hospital with a 4-week history of high fever, productive cough and malaise . Chest roentgenogram showed an infiltrative shadow in the left lower lung field . Isolated bacteria from sputum were acid-fast bacilli and identified as M . szulgai by the DNA-DNA hybridization method . Drug susceptibility tests showed resistance to RFP (MIC>100 microg/ml) . Combined treatment with ethionamide, streptomycin and isoniazid based on the results of drug susceptibility tests resulted in clinical and radiologic improvement within two years . Additional treatment with CAM for another year resulted in complete eradication of the mycobacterium. J Oral Maxillofac Surg, 2000 May, 58(5), 477 - 81 Diagnosis and treatment of cervical tuberculous lymphadenitis; Weiler Z et al.; PURPOSE: This study presents the long-term results of treatment of a series of patients with tuberculous mycobacterial lymphadenitis of the head and neck . PATIENTS: Twenty-one patients were seen in a 10-year period . The median age at onset was of 41.2 years (range, 4 to 79 years), and the male-to-female ratio was 11:10 . Sixteen patients were of Ethiopian origin, 3 from the former USSR, and 2 were Israeli women (1 of Indian and 1 of Morrocan origin) . Symptoms started between 2 weeks and 6 months before presentation (mean, 5.8 weeks) . Most patients had negative chest radiographs, a variable response to the tuberculin skin test, and a negative culture for mycobacterial organisms . RESULTS: Fine-needle aspiration (FNA) of the cervical lymph nodes was the most reliable method to confirm the bacteriologic agent causing the lymphadenopathy . Acid-fast bacilli smears of the aspirate were positive in all but 3 patients, whereas histologic examination of the lymph nodes gave diagnostic results in only two thirds of cases examined . All patients were treated with antituberculous chemotherapy . Sixteen patients also underwent surgical excision of their cervical lymph nodes, and all of them showed a complete response to the combined treatment . The remaining patients reacted to chemotherapy alone with complete cure . One patient died of gastric carcinoma, and the only acquired immune deficiency syndrome (AIDS) patient died a year later of cytomegalovirus encephalitis . CONCLUSION: The most reliable indicator of cervical mycobacterial infection is an acid-fast smear from the FNA specimen . Antituberculous chemotherapy, with or without surgical excision of the involved cervical lymph nodes, is the method of choice for treatment of this disease. Am J Clin Pathol, 2000 May, 113(5), 703 - 8 Pathologic findings for bacille Calmette-Guérin infections in immunocompetent and immunocompromised patients; al-Bhlal LA; The pathologic findings from biopsy specimens from 9 patients with postvaccination bacille Calmette-Guerin (BCG) infection are presented . The patients were vaccinated with BCG during the first 2 days of life . Four patients had normal immunity and 5 patients were immunocompromised . The pathologic findings in both groups were different . Biopsy specimens from patients with normal immunity showed multiple epithelioid granulomas and Langhans giant cells with or without suppuration . Caseous necrosis was minimal . Ziehl-Neelsen stain for acid-fast bacilli showed a few bacilli in 2 cases and was negative in the remaining 2 cases . Biopsy specimens from the second group of patients, who were immunosuppressed, consisted mainly of skin and subcutaneous tissue . These revealed diffuse infiltrates of histiocytes with plump nuclei and abundant "dirty" grayish cytoplasm, which was full of numerous acid-fast bacilli . The clinical course for the 2 groups also was different . Patients with normal immunity generally recover completely, spontaneously or after excision of the suppurative lymph node and usually do not require antibiotic chemotherapy . In immunosuppressed patients, disseminated BCG infection, which may prove fatal, may develop . These patients should receive a full course of antituberculous chemotherapy and, in addition, treatment of the underlying immunologic disorder. Microb Pathog, 2000 May, 28(5), 313 - 8 Secretion of cytokines by human macrophages upon infection by pathogenic and non-pathogenic mycobacteria; Beltan E et al.; The interaction of various pathogenic (Mycobacterium tuberculosis, M . avium, M . kansasii, M . xenopi), and non-pathogenic mycobacteria (M . smegmatis, M . phlei) with human macrophages at the level of macrophage cytokine expression (TNFalpha, IL1, IL6 and GM-CSF) was investigated . Both for TNFalpha and GM-CSF, the lowest levels were obtained with pathogenic mycobacterial species, whereas about 2-8 times higher levels were observed for non-pathogenic species . Contrary to the above, the differences for IL6 and IL1 were not marked, although IL6 appeared to be more elevated for non-pathogenic species . Heat-killed bacteria induced a lower level of the cytokines for all the three cytokines assayed (TNFalpha, IL6 and IL1), except for M . tuberculosis for whom a significantly higher proportion of TNFalpha was induced by killed bacilli . The RT-PCR experiments performed on M . avium (as a low inducer of the cytokines) and M . smegmatis (as a high inducer of the cytokines) showed that the differences observed among pathogenic vs non-pathogenic strains were also reflected at the transcriptional level for TNFalpha and to a lesser extent for IL6, but not for IL1 . This investigation underlined important differences existing between the pathogenic and non-pathogenic species, particularly as regards TNFalpha and GM-CSF . Indian J Pediatr, 1999 Jan-Feb, 66(1), 148 - 50 Congenital tuberculosis; Balasubramanian S et al.; A 29 day old male infant presented with a history of fever, cough, increasing respiratory distress and abdominal distension from the 10th day of life . Examination revealed failure to thrive, marked tachypnea, pallor, hepatosplenomegaly and harsh vesicular breath sounds . Chest skiagram showed extensive broncho-pneumonic changes . As the infant did not respond to antibiotics, he was investigated for tuberculosis . The gastric aspirate smear showed plenty of acid fast bacilli (AFB) and culture showed M . tuberculosis growth . Endometrial biopsy of the mother showed tuberculous granuloma and acid fast bacilli and culture of the aspirate from endometrium grew M . tuberculosis . The need for endometrial biopsy of mothers of infants with congenital tuberculosis is highlighted. Dakar Med, 1999, 44(1), 28 - 31 {Pneumocystosis in HIV infected patients presenting with acid-fast bacilli negative pneumopathy at the Central University Hospital at Dakar}; Dieng Y et al.; Pneumocystosis is an opportunist parasitic disease which occurs currently at Europe and United States in HIV infected patients . In Africa, the disease is not current . Pneumocystosis has been detected in the Fann medical universitary center at Dakar from HIV infected patients with acido alcoholo resistant bacilli negative pneumopathy . Analysis of broncho alveolar liquid(BAL) of 29 patients after Giemsa and Blue of Toluidin O staining allowed isolating of two cases of pneumocystosis . A man and a woman were the patients . They were HIV1 positive with at X ray bilateral interstitial syndrome . The CD4 lymphocytes count of the one was lower than 200/mm3 and for the other it was higher than 200/mm3. Diagn Microbiol Infect Dis, 2000 May, 37(1), 31 - 6 Evaluation of the BACTEC MGIT 960 system for the recovery of mycobacteria; Kanchana MV et al.; We evaluated the BACTEC MGIT 960 system, which is a fully automated, non-invasive, continuous monitoring system for the growth and detection of mycobacteria . Including respiratory and other specimens, 1,742 specimens were processed and inoculated into the BACTEC MGIT 960 and the BACTEC 460 TB Systems, as well as onto Lowenstein-Jensen (L-J) media . A total of 104 isolates of mycobacteria were recovered from all culture systems . This included Mycobacterium tuberculosis complex, Mycobacterium avium-intracellulare (MAI) complex and other mycobacteria (MOTT) . The isolation rates for M . tuberculosis complex and MAI complex were comparable for the BACTEC 460 (54.8% and 13.5%) and the BACTEC MGIT 960 (51.9% and 13.5%) . The overall isolation rate was less for BACTEC MGIT 960 (76.9%) which was due to lesser number of MOTT isolates recovered from this system . The mean times to detection (TTD) for all mycobacteria were 9.3 days for the BACTEC MGIT, 14.6 days for the BACTEC 460 and 21.6 days for L-J . A significant difference was observed when TTD was tested in relation to degree of positivity in smears, with the BACTEC MGIT maintaining the short TTD even with less number of bacilli in the smear . The contamination rates were, 6.4% for BACTEC MGIT, 2.9% for BACTEC 460 and 12.1% for L-J medium . The BACTEC MGIT 960 system shows performances comparable to the BACTEC 460 and seems to be a dependable, user friendly system. Scand J Immunol, 2000 May, 51(5), 502 - 10 Mechanisms involved in protective immune response generated by secretory proteins of Mycobacterium habana against experimental tuberculosis; Divya Jyothi M et al.; Live mycobacteria secrete a number of unique proteins early in their multiplication which are important for both the pathogenesis and the stimulation of specific host responses . We have investigated the mechanisms by which the host mounts immune response against tuberculosis after vaccination with secretory proteins (SP) of a vaccine candidate Mycobacterium habana TMC 5135 . Mice vaccinated with SP of 10th day growth of M . habana, either alone or emulsified in Freund's incomplete adjuvant (FIA) possessed antituberculous resistance and cellular immune responses against M . tuberculosis H37Rv . These proteins induced a significant cutaneous delayed type hypersensitivity response in guinea pigs vaccinated with heat killed M . tuberculosis H37Rv, which was equivalent to that observed with a standard purified protein derivative (PPD) . The splenocytes of these guinea pigs have shown higher proliferative response after stimulation with SP than with PPD . The SP + FIA immunization has been found to exert maximum prophylactic effect by potentiating both the oxygen dependent arms and enzymatic activities of macrophages . Macrophages from mice vaccinated with SP of M . habana produced enhanced levels of interleukin(IL)-2, interleukin-12 and interferon(IFN)-gamma . The protective as well as cell mediated immune responses were upregulated in SP immunized animals when compared to whole cell (M . habana) vaccinated animals . SDS-PAGE of SP from M . habana showed the prominent bands of 60, 32, 31 and 30 kDa . Furthermore, the western analysis of SP with pulmonary tuberculosis patient's serum has revealed the presence of immunoreactive antigens of 36, 35, 33/32 kDa . Overall study demonstrated that the secretory antigens released by actively growing M . habana bacilli could activate different arms of effective immune response. Pediatr Dermatol, 2000 Mar-Apr, 17(2), 133 - 5 Disseminated lupus vulgaris and papulonecrotic tuberculid: case report; Senol M et al.; The incidence of tuberculosis and extrapulmonary forms of this disease is increasing all over the world . Lupus vulgaris is the most prevalent form of cutaneous tuberculosis in Europe and the Middle East . Papulonecrotic tuberculid, the most common form of hyperergic response to mycobacteria or their fragments, is uncommon in children . We report lupus vulgaris with papulonecrotic tuberculid in a 12-year-old girl who had a 3-year history of slowly growing plaques on her trunk, extremities, and the tip of her nose and papuloulcerative lesions over her entire body . A skin biopsy specimen showed minimally caseating granulomatous inflammation . Staining for acid-fast bacilli was negative in both plaques and papules . Polymerase chain reaction identified Mycobacterium tuberculosis DNA in the patient's sputum, gastric fluid, and plaques and was negative in the papules . She was started on antituberculous therapy with four drugs and her lesions responded rapidly. Lett Appl Microbiol, 2000 May, 30(5), 385 - 9 Effect of ammonia on the growth of Bacillus species and some other bacteria; Leejeerajumnean A et al.; The tolerance of 26 Bacillus species isolated from alkaline fermented foods, five other bacilli and nine non spore-forming bacteria to alkaline pH and ammonia was determined . All grew at pH 7, 8 and 9 in the presence of 930 mmol l-1 NH4 + at pH 7.0, and in the presence of NH3 concentrations up to 5 mmol l-1 at pH 7.0 and 8.0 . At higher NH3 concentrations, growth of some of the bacteria was inhibited and at 500 mmol l-1 only B . pasteurii and B . pumilus grew . Bacteria from alkaline food fermentations included strains relatively sensitive to NH3 (inhibited by 50 mmol l-1) and relatively tolerant strains (grew in the presence of 300 mmol l-1) and there was no evidence that they were more tolerant to NH3 than bacteria not associated with these fermentations. J Appl Microbiol, 2000 Apr, 88(4), 669 - 77 The physiology and pathogenicity of Mycobacterium tuberculosis grown under controlled conditions in a defined medium; James BW et al.; A chemically-defined culture medium was developed which supported batch growth of Mycobacterium tuberculosis, strain H37Rv, at a minimum doubling time of 14.7 h . This medium also facilitated chemostat culture of M . tuberculosis at a constant doubling time of 24 h . Chemostat growth was optimized at a dissolved oxygen tension of 20% (v/v) and 0.2% (v/v) Tween-80 . Chemostat cultures were dispersed suspensions of single bacilli (1.5-3 microm long), or small aggregates, at a mean density of log10 8.3 cfu ml-1 . A limited number of amino acids was utilized (alanine, asparagine, aspartate and serine were depleted by >50%; glycine, arginine, isoleucine, leucine and phenylalanine, by approximately 40%) . Chemostat-grown cells were pathogenic in aerosol-infected guinea pigs, producing disseminated infection similar to that caused by plate-grown cells . Cells from chemostat culture were significantly more invasive for J774A.1 mouse macrophages than agar- or batch-grown cells . This study demonstrates the suitability of chemostat culture for the growth of pathogenic mycobacteria in a defined physiological state with potential applications for the controlled production of mycobacterial components for therapeutic and vaccine applications. Lupus, 2000, 9(2), 147 - 50 Intramedullary tuberculosis manifested as Brown-Sequard syndrome in a patient with systemic lupus erythematosus; Kim WU et al.; A 25-year-old girl presented with progressive deterioration of right side weakness with decreased sensation on the left trunk . She had been treated with high dose steroid due to autoimmune thrombocytopenia for 2 months . Clinical, laboratory and immunologic studies revealed that she had systemic lupus erythematosus (SLE), MRI of spinal cord showed marginal contrast enhancing and fluid containing mass in the cord of the C5-6 level, suggesting intramedullary abscess . She underwent surgery of mass removal with biopsy . The pathologic findings from cord tissues revealed numerous acid fast bacilli (AFB) in necrotic tissues . After surgery and anti-tuberculous treatment, her neurologic symptoms were markedly improved with restoration of right side motor weakness . To our knowledge, this is the first case report of intramedullary tuberculosis in a patient with SLE . Since intramedullary tuberculosis may sometimes mimic neurologic complication of SLE itself, it may pose diagnostic and therapeutic confusion for clinicians . We report a case of spinal cord tuberculosis affecting C5, 6 level which was manifested as Brown-Sequard syndrome in a patient with SLE. Kansenshogaku Zasshi, 2000 Mar, 74(3), 245 - 9 {Why there were few isolations of tubercle bacilli in young children with pulmonary tuberculosis?}; Ibe M et al.; We retrospectively evaluated the factors influencing the isolation of tubercle bacilli in 51 children under 14 years of age with pulmonary tuberculosis who were admitted to Yokohama City University Hospital from 1975 to 1998 . Young children (0-6 years of age) with pulmonary tuberculosis were significantly less positive by smear and culture than elder children (7-14 years of age) with pulmonary tuberculosis . According to the Japanese Society for Tuberculosis classification of finding on chest X-ray film for pulmonary tuberculosis, the culture-positive patients with type II (cavitary lesions) were found in all, the culture-positive patients with type III (non-cavitary lesions) in 39.3%, and the culture-positive patients with type H (hilar and mediastinal lymphadenopathys) in 35.3% . Patients with cavitary lesions (type II) were significantly more positive by smear and culture than patients with non-cavitary lesions (type III + type H) . Only 15.8% of the young children with pulmonary tuberculosis had received BCG vaccine and all had non-cavitary lesions (type III and type H) . But, 84.6% out of the older children had received BCG vaccine and half had cavitary lesions . Taken together, the result was that there were few isolation of tubercle bacilli in young children with pulmonary tuberculosis because they had non-cavitary tuberculosis without delayed-type hypersensitivity to tubercle bacilli. Indian Pediatr, 2000 Apr, 37(4), 375 - 82 Polymerase chain reaction in the diagnosis of tuberculosis; Jatana SK et al.; OBJECTIVES: To evaluate efficacy of polymerase chain reaction (PCR), using the insertion sequence IS6110 as target for DNA, to detect Mycobacterium tuberculosis in body fluids of children with suspected tuberculosis (TB) . SETTING: Hospitalized patients . METHODS: A comparison of PCR on body fluids, Acid Fast Bacilli staining (AFB), mycobacterial culture and clinical features, with special emphasis on central nervous system (CNS) TB was done over 18 month period . A total of 80 children were evaluated, 41 with probable TB disease and 39 controls . Cases were defined by specific clinical criteria . Controls included patients free of clinical TB . PCR was done on the clinical specimens and compared with clinical findings, radiological features, Mantoux (Mx) testing, AFB staining and culture on Lowenstein-Jensen (LJ) medium . RESULTS: Sensitivity of PCR in CSF samples was 100%, in gastric aspirate samples was 20% and in pleural fluid samples was 100% . CONCLUSION: PCR technique may become a valuable diagnostic tool for the diagnosis of tuberculosis in children especially in CNS TB. Haemophilia, 2000 Mar, 6(2), 66 - 70 The use of central venous catheters (portacaths) in children with haemophilia; Bollard CM et al.; The experience with central venous implantable devices (portacaths) has been reviewed in children attending the Auckland Hospital Haemophilia Centre . Fourteen children had 23 portacaths inserted . Thirteen had severe Haemophilia A, of whom five had high responding inhibitors to factor VIII . All the children were HIV negative . Ages ranged from 4 months to 13 years at the time of initial placement and 12 were under 5 years . Indications for portacath placement included primary and secondary prophylaxis, induction of immune tolerance, prophylactic therapy post intracranial haemorrhage and poor venous access . Catheter-related infections occurred in 48% of cases . Staphylococcal species were the most common organisms isolated followed by gram-negative bacilli . 63% of the infections were successfully cleared with antibiotics . Haematoma formation occurred in 17% of catheters, primarily in patients who had high factor VIII inhibitor levels . Mechanical problems including blockage, leakage and extrusion of the portacath occurred less frequently (13%) . The significant rate of infection in this immunocompetent population is consistent with other reports . Despite the obvious benefits of portacaths this complication is potentially serious and causes appreciable morbidity . In contrast, bleeding complication rates were relatively low. Int J Tuberc Lung Dis, 2000 Apr, 4(4), 371 - 6 Bleach sedimentation method for increased sensitivity of sputum smear microscopy: does it work? Van Deun A, Maug AK, Cooreman E, Hossain MA, Chambuganj N, Rema V, Marandi H, Kawria A, Portaels F. SETTING: A non-governmental organisation (NGO) supported tuberculosis control programme in Bangladesh with good smear microscopy . OBJECTIVE: To verify whether bleach sedimentation method increases the sensitivity of sputum smear microscopy for acid-fast bacilli (AFB), and if so, how . DESIGN: Duplicate smears from successive routine specimens, peripheral centres examining direct smears, and blind examination of bleach sediment smears at central laboratories . RESULTS: When all 3,287 sputum samples were examined in duplicate and the International Union Against Tuberculosis and Lung Disease cut-off for positivity was applied, more positives were not found by bleach sedimentation . Using the much lower American Thoracic Society (ATS) threshold, the percentage positives rose slightly from 15.5% for direct smear to 16.6% after bleach . The gain was more evident when suspect examinations only were taken into consideration, as bleach missed many positives identified by direct follow-up smear . When patients rather than individual smears were counted, more suspects were detected by bleach (10% gain on average), but with considerable variation between the centres (range 6-16%) . To arrive at this gain, the ATS cut-off was used, with corrections for false results . Under routine conditions, however, this threshold is too low in view of possible transfer of AFB . CONCLUSIONS: Bleach sedimentation can increase the diagnostic yield, but only to a minor extent if all other factors have been optimised already; it is not a panacea . Precautions against false negatives as well as false positives should be taken, and the additional workload is not negligible. Mikrobiologiia, 2000 Mar-Apr, 69(2), 266 - 9 {Effect of chitosan derivatives on the development of phage infection in cultured Bacillus thuringiensis}; Kochkina ZM et al.; The influence of chitosan fragments with different degrees of polymerization and some chemical chitosan derivatives on the infection of Bacillus thuringiensis by phage 1-97A was studied . It was shown that chitosan inhibits phage infection and inactivates phage particles . The extent of inhibition of phage infection inversely depended on the degree of polymerization of chitosan fragments . On the contrary, the extent of inactivation of phage virulence was proportional to the degree of polymerization . Chitosan derivatives did not inhibit the growth of bacilli . Deaminated chitosan derivatives at a concentration of 100 mg/ml efficiently inhibited phage reproduction, exhibiting no correlation between the degree of deamination and antiviral activity . The anionic derivative chitosan sulfate and N-succinate-6-O-sulfate did not inactivate phage, did not influence bacterial growth, and did not inhibit the process of viral infection. Southeast Asian J Trop Med Public Health, 1999 Sep, 30(3), 539 - 41 Prevalence of communicable disease among restaurant workers along a highway in Assam, India; Biswas D et al.; 105 workers of 29 roadside restaurants along a highway in Assam were interviewed for their habits, educational status and were examined for their hygienic status and presence of communicable diseases (STDs,TB etc) . Most of them were young, males with 40% literacy; more than half were unmarried . About 30% of them were alcoholic and smokers and 2.9% were addicted to cannabis . The hygienic conditions of the workers were poor . More than one third had sexual contact with multiple sex partners or commercial sex workers and 2% were engaged in homosexual activity . Most of them did not use condom . 25.7% of them had genital lesions suggestive of sexually transmitted disease, 11.8% showed gram negative diplococci in urethral smears and 5.1% were VDRL reactive . Skin infections followed by gastrointestinal disorders and respiratory tract infections were other prevalent problems . 70.6% were positive for intestinal parasites and 22.2% were sputum positive for acid fast bacilli . However all the collected blood samples were negative for HIV. Southeast Asian J Trop Med Public Health, 1998 Dec, 29(4), 860 - 3 The value of fluorescence microscopy of auramine stained sputum smears for the diagnosis of pulmonary tuberculosis; Singh NP et al.; Laboratory diagnosis of pulmonary tuberculosis rests on the bacteriological examination of sputum smears stained by the Ziehl-Neelsen (ZN) method for acid fast bacilli (AFB) . In the present study, we have compared light microscopy of ZN stained smears with that of fluorescence microscopy of sputum smears stained by auramine-phenol flurochrome dye for detection of AFB in sputum specimens . Sputum specimens from a total of 2,600 clinically suspected and diagnosed cases of pulmonary tuberculosis were examined by both the methods . Sputum specimens from a total of 1,104 patients were found to be positive for AFB . These included sputa from 975 (37.5%) patients positive for AFB by both ZN and auramine staining methods and sputa from an additional 129 (4.96%) patients positive for AFB by auramine staining only . Thus auramine staining of sputum smears in comparison to that of ZN staining is a better method of sputum microscopy for demonstration of AFB in sputum specimens . Fluorescence microscopy is relatively more sensitive and has the added advantage of allowing a large number of sputum specimens to be examined in a given time, in laboratories equipped with a fluorescent microscope. Biochem J, 2000 May 1, 347 Pt 3, 725 - 31 The domains of human fibronectin mediating the binding of alpha antigen, the most immunopotent antigen of mycobacteria that induces protective immunity against mycobacterial infection; Naito M et al.; We have recently shown that alpha antigen (alpha-Ag), the immunodominant antigen of mycobacteria, has a novel fibronectin (FN)-binding motif that is unique among mycobacteria {Naito, Ohara, Matsumoto and Yamada (1998) J . Biol . Chem . 273, 2905-2909} . In this study, we examined the domains of human FN that interacted with alpha-Ag . Fragments of FN generated by either proteolysis or recombinant DNA techniques were compared for their ability to bind to alpha-Ag . Fragments containing either the C-terminal heparin-binding domain or the central cell-binding domain consistently bound to alpha-Ag . The fragment of the C-terminal heparin-binding domain, upon mutation that resulted in the loss of its heparin-binding activity, could not bind with alpha-Ag . These findings suggested that the mutated site, i.e . the main heparin-binding site of FN, was also the principal site for binding to alpha-Ag . The alpha-Ag-binding domains of FN could bind whole mycobacterial bacilli, suggesting that these two domains are important contributors to mycobacterial infection. J Acquir Immune Defic Syndr, 1999 Nov 1, 22(3), 302 - 7 Gender difference in skin reactivity to purified protein derivative among carriers of HTLV-I in Japan; Hisada M et al.; The incidence of malignancies due to oncogenic virus infections tends to be higher in men than in women . Gender-related differences in cell-mediated immunity, which plays a role in viral pathogenesis, may explain this observation . To explore this possibility in the context of HTLV-I infection, we examined skin reactivity to purified protein derivative (PPD) among 128 residents of an HTLV-I endemic area in Japan, who were born before 1921 and are assumed to have been exposed to M . tuberculosis bacilli . The odds ratio (OR) for reduced PPD reactivity (erythema <10 mm in diameter) was calculated by multiple logistic regression analysis . Men were significantly less likely than women to have reduced PPD reactivity among HTLV-I-negative individuals (26% versus 59%; p < .01); whereas this gender difference was not apparent among HTLV-I carriers (63% versus 62%; p = .87) . HTLV-I positivity was strongly associated with reduced PPD reactivity in men, but not in women (odds ratio {OR}, 7.3 versus 1.2; p = .05) . Although this observation may be due, in part, to a longer average duration of HTLV-I infection in men compared with women, the finding also raises the possibility that men may be inherently more susceptible to loss of PPD reactivity by HTLV-I infection. Infect Immun, 2000 May, 68(5), 2854 - 62 Capsular polysaccharide and O-specific antigen divergently modulate pulmonary neutrophil influx in an Escherichia coli model of gram-negative pneumonitis in rats; Russo TA et al.; Enteric gram-negative bacilli cause a severe, often life-threatening pneumonia . An improved understanding of the pathogenesis of this infection may lead to improved treatment . Nearly all of the responsible gram-negative bacilli possess capsular polysaccharides and/or an O-specific antigen as part of their lipopolysaccharide (LPS) . We hypothesized that these surface polysaccharides may modulate the pulmonary host response . To investigate this, a rat pneumonitis model was used, and pulmonary neutrophil influx, a critical aspect of host defense, was measured . To assess for the effect of the capsule and O-specific antigen on this host response, three proven, isogenic derivatives that are deficient in capsular polysaccharide alone (CP9.137), the O-specific antigen moiety of the LPS alone (CP921), and both the capsular polysaccharide and O-specific antigen (CP923), as well as their wild-type parent (CP9), were used as challenge strains at various intratracheal challenge inocula (CI) . Total lung myeloperoxidase (MPO), a surrogate marker for neutrophils, was measured for 15 h post-bacterial challenge . To determine the effect of capsule and the O-specific antigen on the measured MPO levels, a mathematical model was developed and used to describe the MPO levels as a function of time for each CI of each of the four strains . The results from this analysis demonstrated that in the absence of the K54 capsule, 80.7 times the CI is necessary to achieve the same maximum MPO level relative to K54 positive strains (P < 0.0001) . In contrast, a diametric effect was observed in the absence of the O-specific antigen, where 0.13 times the CI was necessary to achieve the same maximum MPO level relative to O4-positive strains (P = 0.0032) . No interactive effect was observed between the capsule and the O-specific antigen . These findings demonstrate that these surface polysaccharides modulate pulmonary neutrophil influx and suggest that the K54 capsular polysaccharide is a proinflammatory mediator and that the O4-specific antigen attenuates the proinflammatory response . If these speculations are substantiated, an understanding of how the capsule and the O-specific antigen modulate host response could have significant therapeutic implications . The potential use of biologic modulators directed against the host response, as well as approaches based on inactivating bacterial components (e.g., surface polysaccharides) in attempts to modify sepsis syndromes, could be developed. Am J Respir Crit Care Med, 2000 Apr, 161(4 Pt 1), 1076 - 80 Predicting the outcome of therapy for pulmonary tuberculosis; Wallis RS et al.; Patients vary considerably in their response to treatment of pulmonary tuberculosis . Although several studies have indicated that adverse outcomes are more likely in those patients with delayed sputum sterilization, few tools are available to identify those patients prospectively . In this study, multivariate models were developed to predict the response to therapy in a prospectively recruited cohort of 42 HIV-uninfected subjects with drug-sensitive tuberculosis . The cohort included 2 subjects whose initial response was followed by drug-sensitive relapse . The total duration of culture positivity was best predicted by a model that included sputum M . tuberculosis antigen 85 concentration on Day 14 of therapy, days-to-positive in BACTEC on Day 30, and the baseline radiographic extent of disease (R = 0.63) . A model in which quantitative AFB microscopy replaced BACTEC also performed adequately (R = 0.58) . Both models predicted delayed clearance of bacilli in both relapses (> 85th percentile of all subjects) using information collected during the first month of therapy . Stratification of patients according to anticipated response to therapy may allow TB treatment to be individualized, potentially offering superior outcomes and greater efficiency in resource utilization, and aiding in the conduct of clinical trials. Am J Otolaryngol, 2000 Mar-Apr, 21(2), 122 - 6 Laryngeal tuberculosis; Yencha MW et al.; Since the introduction of antituberculous medications, the incidence of laryngeal tuberculosis (TB) has decreased and remains stable . However, with the incidence of TB increasing, mainly caused by the acquired immunodeficiency syndrome epidemic, the incidence of laryngeal involvement may be on the rise . The main presenting symptom of laryngeal TB is dysphonia . The diagnosis is confirmed with the identification of granulomatous inflammation, caseating granulomas, and acid-fast bacilli on histopathologic examination of biopsied laryngeal tissue . However, making the diagnosis difficult can be the presence of pseudoepitheliomatous hyperplasia, which mimics squamous cell carcinoma . Treatment is primarily with antituberculous medications with surgery reserved for those cases of airway compromise . Laryngeal complications can occur; thus, long-term follow-up is recommended . We report a case of laryngeal TB in a human immunodeficiency virus-negative patient and review the literature. Rev Med Chir Soc Med Nat Iasi, 1999 Jul-Dec, 103(3-4), 206 - 9 {Gram-negative anaerobic bacilli identified in the subgingival microflora associated with fixed dentures}; Stadoleanu C et al.; The prosthodontics devices have a major role in healthy state or disease of periodontal structures . They may initiate or amplify tissues destruction when technical features or the used material are not the most appropriate . The aim of this study was to establish the influence of different materials on the evolution of chronic periodontal diseases by comparison to lesions associated with natural teeth . Lot of study was represented by 21 partial edentulous patients with periodontitis in both natural teeth and fixed prosthodontics . Periodontal pockets were sampled and then Gram stained smear and cultivation onto selective media (Schadler + 5% sheep blood + nalidixic acid + vancomycin) were done . The isolated bacteria were identified by standard methods . We observed quantitative and qualitative differences between studied patients according to the material used for making the fixed prosthodontics. Int J Tuberc Lung Dis, 2000 Mar, 4(3), 272 - 4 Viability of stored sputum specimens for smear microscopy and culture; Banda HT et al.; A laboratory study was performed to determine how long sputum specimens from smear-positive tuberculosis patients can be stored at room temperature or in the refrigerator and retain a positive acid-fast bacilli (AFB) smear or a positive mycobacterial culture . Sputum samples from 30 patients were examined up to 4 weeks and samples from 13 patients examined up to 8 weeks . Provided samples had not dried out, all sputum smears remained AFB positive up to 4 and 8 weeks . In both patient groups, at 4 weeks 37-39% of specimens at room temperature grew mycobacteria compared with 54-67% of specimens stored in the refrigerator . These results have implications for tuberculosis programme policy. Int J Tuberc Lung Dis, 2000 Mar, 4(3), 246 - 51 An incremental cost-effectiveness analysis of the first, second and third sputum examination in the diagnosis of pulmonary tuberculosis; Walker D et al.; SETTING: St . Francis Hospital in Katete District, Eastern Province, Zambia . OBJECTIVE: To compare the incremental cost-effectiveness of examining serial sputum smears for screening suspects for pulmonary tuberculosis at a rural district hospital in Zambia . DESIGN: An incremental cost-effectiveness analysis of serial sputum smear examinations for diagnosing pulmonary tuberculosis based on laboratory results collected during 1997 and 1998 in a rural district hospital in Zambia . The cost analysis took a health service provider perspective, and used the ingredients approach . The cost-effectiveness is expressed in terms of the incremental cost per tuberculosis case diagnosed . Relevant information was obtained from various sources, including administrative records, interviews and direct observation . RESULTS: Of a total of 166 acid-fast bacilli positive suspects who had three sputum smears examined sequentially, 128 (77.1%) were found on the first smear, a further 25 (15%) on the second smear and 13 (7.9%) additional cases were identified on the third smear . The economic analysis shows that the incremental cost of performing a third test, having already done two, increases rapidly with only a small gain in terms of additional cases of tuberculosis identified . CONCLUSION: A policy of examining two samples should be considered in resource-poor settings, if the remaining steps of the national diagnostic algorithm can be adhered to with respect to smear-negative suspects. Mol Pathol, 1999 Oct, 52(5), 283 - 8 PCR based detection of mycobacteria in paraffin wax embedded material routinely processed for morphological examination; Frevel T et al.; BACKGROUND: The incidence of mycobacterial infections has increased during the past five years . A prompt diagnosis is indispensable for initiating appropriate treatment . Because culturing of mycobacteria takes three to six weeks and sensitivity of microscopic detection of acid fast bacilli is low, amplification methods provide promising possibilities . Recently, the polymerase chain reaction (PCR) has been shown to be useful for confirming a mycobacterial infection, especially in cases with unexpected histological findings or lack of suitable material for culturing . AIMS: To evaluate the impact of PCR based techniques in the detection of mycobacterial infections in uncultured routine histological specimens as an alternative to surgical pathology . METHODS: Two hundred and twenty nine formalin fixed and paraffin wax embedded samples from 141 patients with clinical or histological suspicion of a mycobacterial infection