Konomi, N., Lebwohl, E., Mowbray, K., Tattersall, I., and Zhang, D. Detection of mycobacterial DNA in andean mummies. J. Clin. Microbiol. 40(12) (2002) 4738–4740.

The identification of genetic material from pathogenic organisms in ancient tissues provides a powerful tool for the study of certain infectious diseases in historic populations. We have obtained tissue samples from the genital areas of 12 mummies in the American Museum of Natural History collection in New York, N.Y. The mummies were excavated in the Andes Mountain region of South America, and radiocarbon dating estimates that the mummies date from A.D. 140 to 1200. DNAs were successfully extracted from all tissues and were suitable for PCR analysis. PCRs were carried out to detect Mycobacterium tuberculosis complex and mycobacteria other than M. tuberculosis (MOTB). M. tuberculosis complex was detected in 2 out of 12 samples, and MOTB were detected in 7 samples. This study confirmed the adequate preservation of genetic material in mummified tissues and the existence of mycobacteria, including M. tuberculosis, in historic populations in South America.—Authors' Abstract

Stearns, A. T. Leprosy: a problem solved by 2000? Lepr. Rev. 73(3) (2002) 215–224.

It is now the year 2001, and in many endemic regions leprosy remains a public health problem by any definition. It is clear that defining leprosy purely by prevalence side-steps some of the real issues. There is still much to do to solve the problem of leprosy. Control programmes require better tests for early diagnosis if leprosy is to be reduced much further. Treatment of the infection and of reactions is still far from ideal, whilst an effective vaccine would be valuable in high-risk regions. Research into the true incidence in each endemic area is essential, and control programs of the future will need a more detailed understanding of the transmission of M. leprae to permit new logical interventions. Leprosy remains a devastating disease. Much of the damage that it inflicts is irreversible, and leads to disability and stigmatization. This is perhaps the greatest problem posed. It is easy to dwell on the successes of the elimination campaign, so diverting attention from those populations of ‘cured’ patients who still suffer from the consequences of infection. Leprosy should be regarded as a problem unsolved so long as patients continue to present with disabilities. WHO has carried out a highly successful campaign in reducing the prevalence of leprosy, and this needs to be acknowledged, but what is happening to the incidence in core endemic areas? Maintaining this success, however, may be an even greater struggle if funding is withdrawn and vertical programmes are absorbed into national health structures. We must take heed of the historian George Santayana, ‘those who cannot remember the past are condemned to repeat it’. We should take the example of tuberculosis as a warning of the dangers of ignoring a disease before it has been fully controlled, and strive to continue the leprosy elimination programmes until there are no new cases presenting with disability. The World Health Organisation has shown that leprosy is an eminently treatable disease, and has prepared the ground. The leprosy elimination campaigns truly are ‘at a height . . . ready to decline.’ Can it be that this is the chance to take leprosy ‘at the flood’? If so, perhaps an extension of the elimination programs beyond the year 2001 would indeed ‘lead to fortune.’—Authors' Abstract

Alcala, L., Ruiz-Serrano, M. J., Perez-Fernandez Turegano, C., Garcia De Viedma, D., Diaz-Infantes, M., Marin-Arriaza, M., and Bouza, E. In vitro activities of Linezolid against clinical isolates of Mycobacterium tuberculosis that are susceptible or resistant to first-line antituberculous drugs. Antimicrob. Agents Chemother. 47(1) (2003) 416–417.

We evaluated 117 isolates of Mycobacterium tuberculosis for susceptibility to linezolid by the proportion and E-test methods. Linezolid showed high in vitro activity, with all the strains inhibited by 1 micro g of the drug per ml. E-test MICs were at least 4 dilutions lower than their equivalents by the standard proportion method.—Authors' Abstract

van Crevel, R., Alisjahbana, B., Lange, W. C. M., de Borst, F., Danusantoso, H., van der Meer, J. W. M., Burger, D., and Nelwan, R. H. H. Low plasma concentrations of rifampicin tuberculosis patients in Indonesia. Int. J. Tuberc. Lung Dis. 6(6) (2002) 497–502.

Setting: Although rifampicin is a key drug in tuberculosis treatment, little is known about its quality and bioavailability in countries endemic for tuberculosis. High drug levels may lead to increased toxicity, while low drug levels may predispose to treatment failure and relapse. Objective: To investigate possible variations in the bioavailability of plasma rifampicin in tuberculosis patients in Indonesia. Design: Plasma concentrations of rifampicin and the rifampicin content of drug formulations in use were measured among 62 non-selected tuberculosis patients in Jakarta, Indonesia. Results: Plasma concentrations of rifampicin were generally low: 70% of patients had 2-hour plasma concentrations (C_max) below 4 mg/liter. No toxic plasma concentrations of rifampicin (>20 mg/liter) were found. The strongest predictive factor for the magnitude of rifampicin concentrations was the drug manufacturer. The rifampicin content of the different drug preparations used was normal (90.5–103.6% of the reference standard). No association was found between low plasma rifampicin concentrations and delayed sputum conversion or treatment failure. Conclusion: The unexpectedly low plasma concentrations of rifampicin in this setting are most likely due to reduced bioavailability of local drug preparations, as the rifampicin content of the drug preparations was normal. The clinical significance of these findings remains to be determined.—Trop. Dis. Bull.

Feng, Z. and Barletta, R. G. Roles of Mycobacterium smegatis D-Alanine: D-Alanine Ligase and D-Alanine Racemase in the mechanisms of action of and resistance to the peptidoglycan inhibitor D-cycloserine. Antimicrob. Agents Chemother. 47(1) (2003) 283–291.

D-Cycloserine (DCS) targets the peptidoglycan biosynthetic enzymes D-alanine racemase (Alr) and D-alanine:D-alanine ligase (Ddl). Previously, we demonstrated that the overproduction of Alr in Mycobacterium smegmatis determines a DCS resistance phenotype. In this study, we investigated the roles of both Alr and Ddl in the mechanisms of action of and resistance to DCS in M. smegmatis. We found that the overexpression of either the M. smegmatis or the Mycobacterium tuberculosis ddl gene in M. smegmatis confers resistance to DCS, but at lower levels than the overexpression of the alr gene. Furthermore, a strain overexpressing both the alr and ddl genes displayed an eightfold-higher level of resistance. To test the hypothesis that inhibition of Alr by DCS decreases the intracellular pool of D-alanine, we determined the alanine pools in M. smegmatis wild-type and recombinant strains with or without DCS treatment. Alr-overproducing strain GPM14 cells not exposed to DCS displayed almost equimolar amounts of L- and D-alanine in the steady state. The wild-type strain and Ddl-overproducing strains contained a twofold excess of L- over D-alanine. In all strains, DCS treatment led to a significant accumulation of L-alanine and a concomitant decease of D-alanine, with approximately a 20-fold excess of L-alanine in the Ddl-overproducing strains. These data suggest that Ddl is not significantly inhibited by DCS at concentrations that inhibit Alr. This study is of significance for the identification of the lethal target(s) of DCS and the development of novel drugs targeting the D-alanine branch of mycobacterial peptidoglycan biosynthesis.—Authors' Abstract

Itokazu, G. S., Fischer, J. H., Manitpisitkul, P., Hariharan, R., and Danziger, L. H. Lack of effect of nizatidine-induced elevation of gastric pH on the oral bioavailability of dapsone in healthy volunteers. Pharmacotherapy 22(11) (2002) 1420–1425.

STUDY OBJECTIVE: To investigate the effect of histamine2 (H2)-receptor antagonist-induced elevation of gastric pH on oral bioavailability of a single dose of dapsone 100 mg. DESIGN: Prospective, randomized, crossover, open-label, single-dose pharmacokinetic study. SETTING: Teaching hospital. PATIENTS: Sixteen men were enrolled in the study; data from 11 subjects were evaluable. INTERVENTIONS: Participants received two treatments separated by at least 14 days. Treatment A consisted of a single dose of dapsone 100 mg. Treatment B consisted of a single dose of dapsone 100 mg plus two doses of oral nizatidine 300 mg administered 3–4 hours apart to maintain gastric pH above 6.0. Plasma samples collected before and up to 120 hours after dapsone administration were analyzed for dapsone and monoacetyldapsone (MADDS) by high-performance liquid chromatography. Pharmacokinetic parameters were determined by noncompartmental analysis. MEASUREMENTS AND MAIN RESULTS: Gastric pH in the first 6 hours after dapsone administration was above 6.0 for a mean ± S.D. of 1.1% ± 2.9% of the time in the absence of nizatidine and 69.5% ± 18.0% of the time during nizatidine therapy. The geometric mean dapsone maximum plasma concentration (Cmax) declined by 13% (p <0.01), and median time to Cmax occurred 2 hours later (p <0.01) with nizatidine coadministration compared with dapsone alone. Inclusion of the 90% confidence interval for the mean Cmax ratio within the equivalence interval of 0.8–1.25 demonstrated the lack of clinical significance for this modest decrease in Cmax. Neither the area under the dapsone plasma concentration-time curve from zero to infinity nor the elimination half-life of dapsone were significantly altered by nizatidine. No clinically significant changes were observed in the pharmacokinetics of MADDS with regard to coadministration of nizatidine. CONCLUSION: Elevation of gastric pH by H2-receptor antagonists, such as nizatidine, does not result in clinically important changes in the rate or extent of oral dapsone absorption.—Authors' Abstract

Kunichika, N., Miyahara, N., Kotani, K., Takeyama, H., Harada, M., and Tanimoto, M. Pneumonitis induced by rifampicin. Thorax 57(11) (2002) 1000–1001.

An 81-year-old man was admitted to hospital with pulmonary Mycobacterium tuberculosis infection and was treated with rifampicin (RFP), isoniazid (INH), and ethambutol (EB). On day 9 he developed fever and dyspnoea. Chest radiographs showed new infiltration shadows in the right lung. Bronchoalveolar lavage (BAL) was performed and increased numbers of lymphocytes were recovered. Drug induced pneumonitis was suspected so the antituberculous regimen was discontinued and methylprednisolone was administered. The symptoms and infiltration shadows improved. INH and EB were reintroduced without any recurrence of the abnormal shadows. T cell subsets in the BAL fluid and a positive lymphocyte stimulation test for RFP suggest that RFP induced pneumonitis may be related to a complex immunological response.—Authors' Abstract

Otten, T. F., Solov'eva, N. S., and Vishnevskii, B. I. [Sensitivity to levofloxacin of various types of non-tuberculosis Mycobacterium] Antibiot. Khimioter 47(6) (2002) 34–37.

Activity of levofloxacin (Tavanic) against 10 species of nontuberculosis of mycobacteria was investigated by indirect method of absolute concentrations on Levenstain-Jensen media (levofloxacin concentration 5 and mcg/mL). The investigation was performed on 71 strains of nontubercolosis mycobacteria: Mycobacterium avium-intracellulare—24 strains, M. fortuitum—17 strains, M. chelonae—10 strains, M. malmoense—13 strains and 6 other species of mycobacteria. Susceptible to critical levofloxacin concentration were 8 species of 10. Resistance to levofloxacin (10 mcg/mL) was estimated for 16.7 per cent of M. avium-intracellulare and 30 per cent of M. chelonei strains. It is concluded that levofloxacin may be a drug of choice for management of mycobacteriosis caused by M. fortuitum, M. kansasii, M. xenopi, M. malmoense, and in the most of cases due to M. avium-intracellulare and M. chelonae.—Authors' Abstract

Park, W. G., Bishai, W. R., Chaisson, R. E., and Dorman, S. E. Performance of the Microscopic Observation Drug Susceptibility Assay in Drug Susceptibility Testing for Mycobacterium tuberculosis. J. Clin. Microbiol. 40(12) (2002) 4750–4752.

The drug susceptibility testing performance of a broth-based method with microscopic reading of bacillary growth, the microscopic observation drug susceptibility (MODS) assay, was compared to that of the reference 7H10 agar method of proportion by using 53 isolates of Mycobacterium tuberculosis from persons at risk for multidrug-resistant TB. For isoniazid (0.1 micro g/ml) and rifampin (2.0 micro g/ml), there was 100% agreement between MODS results read at day 11 and the reference method. Levels of agreement for ethambutol tested at 2.5 and 7.5 micro g/ml were 70 and 58%, respectively. Levels of agreement for streptomycin tested at 2.0 and 6.0 micro g/ml were 77 and 51%, respectively. For isoniazid and rifampin drug susceptibility testing, MODS is as accurate as and more rapid than the reference method.—Authors' Abstract

Rawat, M., Newton, G. L., Ko, M., Martinez, G. J., Fahey, R. C., and Av-Gay, Y. Mycothiol-deficient Mycobacterium smegmatis mutants are hypersensitive to alkylating agents, free radicals, and antibiotics. Antimicrob. Agents Chemother. 46(11) (2002) 3348–3355.

Mycothiol (MSH; 1D-myo-inosityl 2-[N-acetyl-L-cysteinyl]amido-2-deoxy-alpha-D-glucopyranoside) is the major low-molecular-weight thiol produced by mycobacteria. Mutants of Mycobacterium smegmatis mc(2)155 deficient in MSH production were produced by chemical mutagenesis as well as by transposon mutagenesis. One chemical mutant (mutant I64) and two transposon mutants (mutants Tn1 and Tn2) stably deficient in MSH production were isolated by screening for reduced levels of MSH content. The MSH contents of transposon mutants Tn1 and Tn2 were found to be less than 0.1% that of the parent strain, and the MSH content of I64 was found to be 1 to 5% that of the parent strain. All three strains accumulated 1D-myo-inosityl 2-deoxy-alpha-D-glucopyranoside to levels 20- to 25-fold the level found in the parent strain. The cysteine:1D-myo-inosityl 2-amino-2-deoxy-alpha-D-glucopyranoside ligase (MshC) activities of the three mutant strains were 2% that of the parent strain. Phenotypic analysis revealed that these MSH-deficient mutants possess increased susceptibilities to free radicals and alkylating agents and to a wide range of antibiotics including erythromycin, azithromycin, vancomycin, penicillin G, rifamycin, and rifampin. Conversely, the mutants possess at least 200-fold higher levels of resistance to isoniazid than the wild type. We mapped the mutation in the chemical mutant by sequencing the mshC gene and showed that a single amino acid substitution (L205P) is responsible for reduced MSH production and its associated phenotype. Our results demonstrate that there is a direct correlation between MSH depletion and enhanced sensitivity to toxins and antibiotics.—Authors' Abstract

Ridtitid, W., Wongnawa, M., Mahatthanatrakul, W., Punyo, J., and Sunbhanich, M. Rifampin markedly decreases plasma concentrations of praziquantel in healthy volunteers. Clin. Pharmacol. Ther. 72(5) (2002) 505–513.

BACKGROUND AND OBJECTIVE: Praziquantel is extensively metabolized by the hepatic cytochrome P450 (CYP) enzymes. The CYP3A isoforms are likely to be major enzymes responsible for praziquantel metabolism. Rifampin (INN, rifampicin), a potent enzyme inducer of CYP-mediated metabolism (especially CYP2C9, CYP2C19, and CYP3A4), is known to markedly decrease plasma concentrations and effects of a number coadministered drugs. The aim of this investigation was to study the possible pharmacokinetic interaction between rifampin and praziquantel. METHODS: An open, randomized, 2-phase crossover design was used in each study of single or multiple doses. In the single-dose study, 10 healthy Thai male volunteers ingested single doses of 40 mg/kg praziquantel alone (phase 1) or after pretreatment with 600 mg of oral rifampin once daily for 5 days (phase 2). In the multiple-dose study, all participants received multiple doses of 25 mg/kg praziquantel alone (phase 1) or after 5-day pretreatment with 600 mg of oral rifampin once daily (phase 2). Plasma concentrations of praziquantel in each phase were determined by the HPLC method. RESULTS: In the single-dose study, rifampin decreased plasma praziquantel concentrations to undetectable levels in 7 of 10 subjects, whereas praziquantel concentrations were reduced by rifampin to undetectable levels in 5 of 10 subjects in the multiple-dose study. In 3 subjects with measurable concentrations in the single-dose study, rifampin significantly decreased the mean maximum plasma concentration (C(max)) and area under the plasma concentration-time curve from 0 to 24 hours [AUC(0–24)] of praziquantel by 81% (p <.05) and 85% (p <.01), respectively, whereas rifampin significantly decreased the mean C(max) and AUC(0–24) of praziquantel by 74% (p <.05) and 80% (p <.01), respectively, in 5 subjects with measurable concentrations in the multiple-dose study. The mean C(max) and AUC(0–24) of praziquantel in subjects whose praziquantel concentrations could not be detected in the single-dose study (7 subjects) after rifampin pretreatment were reduced by approximately 99% (p <.001) and 94% (p <.001), respectively, and in the multiple-dose study (5 subjects), they were reduced by 98% (p <.05) and 89% (p <.01), respectively. CONCLUSIONS: Rifampin greatly decreased plasma concentrations of single and multiple oral doses of praziquantel to levels lower than that of the minimum therapeutic concentration. Because praziquantel and rifampin are widely used in the treatment of liver flukes (Opisthorchis viverrini) and Mycobacterium tuberculosis, respectively, in Thailand and in some other countries in southeast Asia, the possibility of one drug influencing the pharmacokinetics of the other must be considered. Therefore simultaneous use of rifampin and praziquantel must be avoided in medical practice to optimize the therapeutic efficacy of praziquantel.—Authors' Abstract

Sekar, B., Elangeswaran, N., Jayarama, E., Rajendran, M., Kumar, S. S., Vijayaraghavan, R., Anandan, D., and Arunagiri, K. Drug susceptibility of Mycobacterium leprae: a retrospective analysis of mouse footpad inoculation results from 1983 to 1997. Lepr. Rev. 73(3) (2002) 239–244.

We analyzed the results of mouse foot pad (MFP) tests performed between 1983 and 1997 in our laboratory for the cases referred with clinical suspicion of relapse/drug resistance. A total of 214 cases, with clinical suspicion of relapse/drug resistance were investigated for susceptibility to the drugs of MDT by MFP inoculation. Among 96 inoculations that showed conclusive results, 81 (84%) were fully sensitive to dapsone, suggesting that most of the clinically suspected relapse is due to drug susceptible Mycobacterium leprae. Of the remaining 15 strains (16%) found resistant to dapsone, 13 (87%) were of high grade resistance and one strain each of intermediate grade and low grade dapsone resistance, suggesting that most of the dapsone resistance is secondary in nature. No case of rifampicin resistance was found. Only one case of combined dapsone and unconfirmed clofazimine resistance was found. No other combined multidrug resistance was observed in our analysis.—Authors' Abstract

Sims, E. J., Goyal, M., and Arnold, C. Experimental versus in silico fluorescent amplified fragment length polymorphism analysis of Mycobacterium tuberculosis: improved typing with an extended fragment range. J. Clin. Microbiol. 40(11) (2002) 4072–4076.

Whole-genome fingerprinting fluorescent amplified fragment length polymorphism (FAFLP) data were compared with in silico data for the sequenced strains of Mycobacterium tuberculosis (H37Rv and CDC1551). For this G+C-rich genome, many predicted fragments were not detected experimentally. For H37Rv, only 108 (66%) of the 163 predicted EcoRI-MseI fragments between 100 and 500 bp were visualized in vitro. FAFLP was also used to identify polymorphism in 10 clinical isolates of M. tuberculosis characterized previously by IS6110 typing, examining fragments of up to 1000 bp in size rather than up to 500 bp as was done previously. Five isolates had unique IS6110 profiles and were not known to be epidemiologically related, two isolates were the same single-band IS6110 type but were not known to be epidemiologically related, and the remaining three isolates were epidemiologically related with identical IS6110 profiles. Analysis of fragments in the 500- to 1000-bp range using nonselective primers differentiated better between strains than analysis of fragments in the 50- to 500-bp range using a set of four selective primers. Seventeen polymorphic fragments were identified between 500 and 1000 bp in size compared with nine polymorphic fragments between 50 and 500 bp. Using the 500- to 1000-bp analysis, a level of discrimination similar to that of IS6110 typing was achieved which, unlike the IS6110 typing, was able to differentiate the two M. tuberculosis strains, each of which had only a single copy of IS6110.—Authors' Abstract

Bastuji-Garin, S., Ochonisky, S., Bouche, P., Gherardi, R. K., Duguet, C., Djerradine, Z., Poli, F., and Revuz, J. Incidence and risk factors for thalidomide neuropathy: a prospective study of 135 dermatologic patients. J. Invest. Dermatol. 119(5) (2002) 1020–1026.

Thalidomide is effective in several cutaneous diseases. Peripheral neuropathy is the most important adverse event limiting its use. Its incidence rate and its relation to thalidomide doses remain unclear. We prospectively monitored 135 patients treated with thalidomide for various dermatologic diseases for 2 yrs in order to estimate the annual incidence rate and risk factors for neuropathy. Patients underwent standardized neurologic examination and nerve conduction studies prior to, and regularly during treatment. Risk factors for neuropathy were assessed using a Cox proportional-hazards model. Clinical and electrophysiologic evidence of a thalidomide-induced neuropathy were present in 25.2% of the patients; however, when considering all potential cases, this rate reached 55.6%. The incidence rate was maximal during the first year of treatment (20%). The risk of neuropathy was related to the daily dose whatever the duration of treatment (p <10–3). Considering a daily dose 50 mg per day as reference, the relative risk for thalidomide neuropathy was 8.2 for a daily dose comprised between 50 and 75 mg per day and 20.2 for a daily dose >75 mg per day (p <10–3). No neuropathy occurred for daily doses 25 mg per day. The neuropathy was subclinical in nearly a quarter of patients with such an adverse event. These data confirm the high rate of thalidomide neuropathy and identify the daily dose as the main risk factor. The risk of neuropathy seems to be negligible for doses less than 25 mg per day, whatever the duration of therapy.—Authors' Abstract

Chaudhry, V., Cornblath, D. R., Corse, A., Freimer, M., and Simmons-O'Brien, E., and Vogelsang, G. Thalidomide-induced neuropathy. Neurology 59(12) (2002) 1872–1875.

BACKGROUND: Thalidomide is effective for the treatment of some refractory dermatologic and oncologic diseases. Toxic neuropathy limits its use, as embryopathy can be avoided by contraceptive measures. OBJECTIVE: To describe the clinical, electrophysiologic, and pathologic features of thalidomide-induced peripheral neuropathy. METHODS: Clinical and electrophysiologic examinations were performed in seven patients with thalidomide-induced peripheral neuropathy. Thalidomide was used for graft-vs-host disease, pyoderma gangrenosum, and discoid lupus with dosages ranging from 100 to 1200 mg/day for 5 to 16 months (cumulative dosages of 24 to 384 g). RESULTS: All seven patients had clinical and electrophysiologic evidence of a sensory more than motor, axonal, length-dependent polyneuropathy that presented as painful paresthesias or numbness. Sural nerve biopsies, done in three patients, showed evidence of Wallerian degeneration and loss of myelinated fibers. The symptoms, signs, and electrophysiologic data correlated with total cumulative dose of thalidomide. CONCLUSIONS: Thalidomide induces a dose-dependent sensorimotor length-dependent axonal neuropathy; it should be judiciously used with close neurologic monitoring.—Authors' Abstract

Courtright, P., Daniel, E., Sundarrao, Ravanes, J., Mengistu, F., Belachew, M., Celloria, R. V., and Ffytche, T. Eye disease in multibacillary leprosy patients at the time of their leprosy diagnosis: findings from the Longitudinal Study of Ocular Leprosy (LOSOL) in India, the Philippines and Ethiopia. Lepr. Rev. 73(3) (2002) 225–238.

Existing prevalence surveys do not provide adequate information to estimate the magnitude of ocular pathology or vision loss in leprosy patients. We sought to determine the prevalence of ocular findings and related risk factors in leprosy patients at the time of their disease diagnosis. We also sought to determine if there were geographic differences and whether these were due to different demographic characteristics of the populations. The study was undertaken at Schieffelin Leprosy Research & Training Centre (Karigiri, India), Leonard Wood Memorial Leprosy Institute (Cebu, Philippines), and (for 3 yrs only) ALERT (Addis Ababa, Ethiopia). Newly diagnosed multibacillary (MB) leprosy patients as well as MB cases relapsed after dapsone monotherapy were eligible for enrollment. In each study site, the target population was 300. Standardized examinations were conducted between 1991 and 1998. Patient enrollment included 301 patients in Karigiri, 289 patients in Cebu, and 101 patients in Addis Ababa. The age-adjusted prevalence of blindness (< 6/60 in the better eye) and visual impairment (6/24–6/60) was 2.8% and 5.2%, respectively. Lagophthalmos and leprosy related uveal changes were detected in 3.3% (95% CI 2.0–4.7%) and 4.1% (95% CI 2.4–5.7) of patients, respectively. Overall, 11% (95% CI 8.5–13.2%) of newly enrolled MB patients had potentially blinding leprosy related ocular pathology. Lagophthalmos was associated with increasing age, a short duration between onset and diagnosis, and a previous reaction involving the face. Uveal conditions were associated with increasing age. Overall, eye disease was more common in Indian and Ethiopian patients compared to Filipino patients; however, differences were not significant when controlling for age and clinical (non-ocular) factors. Patients with potentially blinding leprosy related pathology were over three times more likely to have other (hand and foot) disabilities than patients without pathology. Differences in the prevalence of blindness and potentially blinding leprosy related ocular pathology between the sites could be accounted for by the differences in age and other clinical factors of the patients at the different sites. Findings suggest that, even in the face of active leprosy control efforts, around 11% of patients will have potentially blinding pathology at the time of their diagnosis and 2.8% will be blind. If those patients with lagophthalmos or blindness are considered appropriate for referral for more detailed assessment, approximately 4% of newly diagnosed leprosy patients will require active follow-up for eye care; including those with reaction involving the face will result in 9.4% of patients requiring active follow-up. These people are likely to be older, with a reaction involving the face, and/or with other disabilities than those not requiring active follow-up.—Authors' Abstract

Daniel, E., Koshy, S., Rao, G. S., and Rao, P. S. Ocular complications in newly diagnosed borderline lepromatous and lepromatous leprosy patients: baseline profile of the Indian cohort. Br. J. Ophthalmol. 86(12) (2002) 1336–1340.

Aim: To describe ocular manifestations in newly diagnosed borderline lepromatous (BL) and lepromatous leprosy (LL) patients in India. METHODS: Ocular complications, at enrollment, occurring in all new borderline lepromatous and lepromatous leprosy patients detected by active case finding within the geographically defined leprosy endemic area of the Gudiyattam Taluk in India from 1991 to 1997 who consented to ocular examinations every 6 months, during and 5 yrs after treatment with multidrug therapy (MDT), were studied. RESULTS: Orbicularis oculi weakness (4.62%), lagophthalmos (4.20%), ectropion (0.42%), trichiasis (0.84%), blocked nasolacrimal ducts (1.68%), pterygium (11.34%), impaired corneal sensation (53%), corneal opacity (10.5%), corneal nerve beading (1.68%), punctate keratitis (1.26%), keratic precipitates (4.62%), iris atrophy (1.68%), and cataract (12.6%) were ocular complications seen in the 301 lepromatous patients at enrolment. 4.6% had blind eyes. Increasing age was associated with ocular complications. 80% of patients were skin smear acid fast bacilli (AFB) positive. The LL/BL ratio was 1:6.4. 71% had some limb deformity. 44% had only leprosy related ocular complications (LROC), 28% had only general ocular complications (GOC) while 14% had both LROC and GOC. Ocular complications were significantly related to leg deformities. Corneal nerve beading was seen most in LL patients (100%) having high bacterial content. Lagophthalmos and muscle weakness were associated with reversal reactions. CONCLUSIONS: Corneal nerve beading occurs in LL patients with high bacillary count. Patients with reversal reaction are more likely to present with orbicularis oculi weakness and lagophthalmos. Leprosy related ocular complications and general ocular complications are significant problems in newly diagnosed lepromatous patients. Elderly, deformed, skin smear positive, lepromatous patients are associated with increased ocular morbidity and form a group that require acceptable and accessible eye care.—Authors' Abstract

Ghorpade, A. Inoculation (tattoo) leprosy: a report of 31 cases. J. Eur. Acad. Dermatol. Venereol. 16(5) (2002) 494–499.

Thirty-one female patients with leprosy lesions starting over tattoo marks observed over a period of 16 yrs are reported. All the patients belonged to the Chhattisgarh State, which is highly endemic for leprosy. Most of the patients were in the third decade of life. All of them had ornamental tattooing done by roadside tattoo artists, who used unsterile needles for tattooing a large gathering one after another with the same needles. In all of them, the first lesion of leprosy started over a tattoo mark. Twenty-five cases had only single lesion of leprosy exclusively confined to tattoo marks. The duration between tattooing and appearance of first lesion in most of the cases varied from 10 to 20 yrs. Paucibacillary leprosy was the commonest type observed in 29 cases, while two had multibacillary leprosy. The diagnosis was confirmed by histopathology in all cases. The present report supports the hypothesis of transmission of leprosy in these cases through tattooing. To the best of our knowledge, such a large collection of leprosy cases subsequent to tattooing has not been reported so far.—Authors' Abstract

Halim, N. K. and Ogbeide, E. Haematological alterations in leprosy patients treated with dapsone. East Afr. Med. J. 79(2) (2002) 100–102.

OBJECTIVE: To evaluate the hemoglobin concentration (Hb); total white blood cell count (WBC), differential WBC count; platelet count and reticulocyte count in leprosy patients already treated with dapsone. DESIGN: A case-control study. SETTING: Specialist Hospital Ossiomo, which is a Leprosarium and Haematology laboratory, University of Benin Teaching Hospital (UBTH), Nigeria. SUBJECTS: Seventy six leprosy patients (forty males and thirty six females) age range 13–40 yrs on single dose dapsone. RESULTS: The hemoglobin concentration showed a marked decrease while the reticulocyte count was markedly elevated which was suggestive of hemolytic anemia. There was also lymphocytosis in patients during pre and post dapsone therapy. CONCLUSION: Leprosy patients on a dosage of 100 mg dapsone, are prone to hemolytic anemia. Leprosy patients should routinely have their Hb, WBC, platelet count and reticulocyte count determined, while on dapsone therapy in order to ascertain the presence of hemolysis.—Authors' Abstract

Hegazy, A. A., Abdel-Hamid, I. A., Ahmed el, S. F., Hammad, S. M., and Hawas, S. A. Leprosy in a high-prevalence Egyptian village: epidemiology and risk factors. Int. J. Dermatol. 41(10) (2002) 681–686.

BACKGROUND: The epidemiology of leprosy in rural Egypt is unknown. This prospective household survey was conducted in a high-prevalence Egyptian village in order to explore the epidemiologic characteristics of the disease and to determine the possible socioeconomic and HLA genotype risk factors. METHODS: The subjects of the study were the residents of Kafr-Tambul village in the Dakahlia governorate, Egypt. There were 10,503 inhabitants of the village, of whom 9643 (91.8%) had a complete visual skin examination, and suspected leprosy patients were subjected to histopathological examination and slit skin smears. Each household was interviewed to record personal data on family members, family size, education, occupation, crowding index at sleep, social score and source of water supply. Human leukocyte antigen (HLA) class II genotypes were analyzed in all leprosy patients and in a number of both household (N = 124) and non-household (N = 30) contacts. RESULTS: The overall prevalence of clinical leprosy in the village studied was 24.9/10,000 (95%CI = 16.3–37.6). Individuals above the age of 40 yrs were 4 times more likely to develop leprosy (OR = 4, p = 0.01). The degree of education, crowding index at sleep, social score and source of water supply were found to be unlikely to increase the risk of leprosy (p >0.05). The frequencies of HLA-DR2 and -DQ1 were significantly associated with leprosy (OR = 3.33 and 5.4; CI = 0.95–12.07 and 1.08–30.19, respectively, all p <0.05). CONCLUSIONS: Our study provides the first picture of the epidemiology of leprosy in a high-prevalence village in rural Egypt. Leprosy detection campaigns should be initiated and directed towards high-prevalence villages. Provision of leprosy control activities in rural health units is necessary in order to detect new cases. The risk for leprosy is associated with HLA-DR2 and -DQ1 markers, and these markers appear to increase personal susceptibility to leprosy in this village.—Authors' Abstract

Jain S., Reddy, R. G., Osmani, S. N., Lockwood, D. N., and Suneetha, S. Childhood leprosy in an urban clinic, Hyderabad, India: clinical presentation and the role of household contacts. Lepr. Rev. 73(3) (2002) 248–253.

A retrospective case note study was done of children below the age of 14 yrs who attended Dhoolpet Leprosy Research Centre (DLRC) over the decade 1990–1999. The aim of the study was to describe the pattern of clinical presentation, the role of household or near neighbour contacts and the incidence of neuritis and reactions. In all, 3118 leprosy patients were registered during this period, of whom 306 were children [182 (60%) male]; 95 children had a single patch, 159 had five or fewer than five patches and 37 had multiple patches. The youngest case detected was 9 months old. The spectrum of leprosy in these children was: TT 62 (20.3%); BT 203 (66.3%); BB 3 (1%); BL 23 (7.5%); LL 5 (1.6%) and PNL 10 (3.3%). Twenty-nine cases (9.4%) were smear positive. Ninety-one children (29.7%) developed a reaction, 86 type I and five type II. A history of contact was present in 119 (38.8%) cases, family contact in 113 (95%) and other than family in six (5%). Classification of the contact was available in only 60 patients. Among the contacts of the index case, 21 (35%) suffered from PB leprosy and 39 (65%) from MB leprosy. All contacts were from the immediate family. This study shows that childhood leprosy cases continue to present in significant numbers to this outpatient clinic. There is a high level of family contact with leprosy in these cases, strengthening the strategy of screening children in leprosy-affected households. The high incidence of reactions and nerve damage in children emphasizes the importance of early detection and treatment.—Authors' Abstract

Keita, S., Tiendrebeogo, A., Berthe, D., Faye, O., and N'diaye, H. T. [Predictive value of consultation reasons in the diagnosis of leprosy in Bamako (Mali)]. Ann. Dermatol. Venereol. 129(8–9) (2002) 1009–1011.

INTRODUCTION: One of the weak points in the strategy for eliminating leprosy is the poor quality of screening. To overcome this, the World Health Organization (WHO) encourages endemic countries to run campaigns for the elimination of leprosy by circulating educational messages and mobilizing the medical community for early screening of cases. The aim of our study was to identify the motives for consultation with high predictive value for the diagnosis of leprosy and to determine the late diagnosis factors and hence assist the staff on site to improve the results of their leprosy elimination campaigns. PATIENTS AND METHODS: The study consisted, during the second trimester of 1999, in interviewing all the patients consulting for the first time the Marchoux Institute or the units screening for leprosy in the Bamako area. The interview recorded the motives for consultation, the delay before consulting and the reasons for late consulting. To assess their positive predictive value, the motives for consultation were related to the diagnosis retained (leprosy or not). RESULTS: One thousand one hundred and seventy seven patients were interviewed. The motive for consulting, “suspected leprosy,” scored the highest positive predictive value (PPV) (80 p. 100): 12 cases of leprosy were diagnosed by 15 consultants having suspected leprosy. Neurological problems were the second motive for consultation (PPV = 61.9 p. 100). The most frequent motive for consultation was spots or “macules” (20 p. 100 of consultations), but only provided a positive predictive value of 19 p. 100. Prior consultations and non-specialized treatments were identified as factors of delay in diagnosing leprosy (p <0.001). CONCLUSIONS: Diagnosis of leprosy cannot be based on the motives for dermatological consultation alone. The macules are the most apparent signs, but of low predictive value. Nevertheless, they are an early but non-specific sign of leprosy and are often neglected by the patient. Other than macules, attention must be paid to the neurological signs (dysesthesia, motor disorders) when screening for leprosy. These signs may appear early on, or be observed at a late stage in the progression of the disease.—Authors' Abstract

Lawn, S. D., Wood, C., and Lockwood, D. N. Borderline tuberculoid leprosy: an immune reconstitution phenomenon in a human immunodeficiency virus-infected person. Clin. Infect. Dis. 36(1) (2003) E5–E6.

Two months after starting highly active antiretroviral treatment (HAART), an individual with human immunodeficiency virus type 1 (HIV-1) infection and profound CD4+ T lymphocytopenia developed several erythematous plaques on his face, which were due to borderline tuberculoid leprosy with reversal reaction. The temporal association between the development of these lesions and changes in blood CD4+ lymphocyte count and plasma HIV-1 load observed during HAART strongly suggests that the presentation of leprosy resulted from immune reconstitution.—Authors' Abstract

Pan Shu, Pan XiaoFeng, and Liu TongKui. Analysis on nerve impairment of the upper limb in 641 leprosy patients. J. Clin. Dermatol. 31(7) (2002) 424–425.

Out of 1575 patients from Xinghua, Jiangsu, China, with active and non-active leprosy, 641 (40.7%) that had nerve impairment in the upper limbs were included in the study. Lateral nerve impairment was seen in 23.17%, which was higher than the incidence of bilateral nerve impairment (17.52%). Nerve impairment was present in 69.23% of active and relapse cases and in 40.46% of non-active cases. 36.63% involved the ulnar nerve, 16.95% involved the median nerve, and 2.35% involved the radial nerve. Claw hand was seen in 73.03% of the cases. Most of the active and relapse cases had single nerve involvement and two-thirds were irreversible. Nerve involvement differs due to delay in diagnosis, different leprosy reactions and different clinical types.—Tropical Diseases Bulletin

Pattyn, S. and Grillone, S. Relapse rates and a 10-year follow-up of a 6-week quadruple drug regimen for multibacillary leprosy. Lepr. Rev. 73(3) (2002) 245–247.

Between 1989 and 1993, 136 multibacillary leprosy patients received a 6-week treatment regimen consisting of daily rifampicin 600 mg, ofloxacin 400 mg, clofazimine 100 mg and a weekly dose of 100 mg minocycline. A previous analysis after a mean follow-up of 4–7 yrs revealed a relapse rate of 2%, involving six late (after more than 5 yrs of follow-up) relapses. During the following years, 12 more relapses appeared during years 8–9 of follow-up. A mean follow-up period of 5 yrs is insufficient to evaluate treatment regimens in multibacillary leprosy. The present regimen cannot be recommended.—Authors' Abstract

Peters, E. S. and Eshiet, A. L. Male-female (sex) differences in leprosy patients in south eastern Nigeria: females present late for diagnosis and treatment and have higher rates of deformity. Lepr. Rev. 73(3) (2002) 262–267.

A study was undertaken to investigate the possibility that female leprosy patients in South Eastern Nigeria may be at a disadvantage with regard to early presentation for diagnosis and the prevention of disability. A hospital-based retrospective examination of case notes for the period 1988–1997 was undertaken, totalling 2309 adult patients of whom 1527 (66 degrees/a) were male and 782 (33%) were female (confirming the usual 2:1 male : female ratio for this disease). Data were collected on 1) the clinical type of leprosy, 2) the interval between the onset of symptoms or signs and presentation for diagnosis and treatment and 3) the patterns of physical deformity/disability. The results indicate that in this part of Nigeria, female leprosy patients have a much longer period (duration of illness) between first symptoms or signs and presentation for diagnosis, compared with males; on average, the period before diagnosis in women was almost twice as long as that in men. Furthermore, they suffered a higher proportion of disabilities. There was no evidence to support discrimination against females with leprosy by the health staff or community and female health workers were available in both hospital and primary health care centres to receive and examine female patients. The Discussion refers to the many studies already published on gender issues, identifying a wide range of social, cultural and economic variables attributed by social structure to men and women, and including the impact of stigma, which may be particularly damaging to women in some situations. The main factors that account for late presentation of females with leprosy in this area have however still to be defined. The consequent higher proportion of disability/deformity in women is obviously of considerable concern, underlining the need for further clinical and social research in this part of Nigeria.—Authors' Abstract

Reichart, P. A., Samaranayake, L. P., Samaranayake, Y. H., Grote, M., Pow, E., and Cheung, B. High Oral Prevalence of Candida krusei in Leprosy Patients in Northern Thailand. J. Clin. Microbiol. 40(12) (2002) 4479–4485.

Although Candida albicans is the most common human yeast pathogen, other Candida species such as C. krusei are now recognized as emerging agents, especially in patients with human immunodeficiency virus (HIV) disease. C. krusei is inherently resistant to the widely used triazole antifungal fluconazole and poses therapeutic problems, especially in systemic candidiasis. In a surveillance study of leprosy patients (with arrested or burnt-out disease) in a leprosarium in northern Thailand, we found a rate of oral carriage of C. krusei (36%) significantly (p <0.05) higher than that for a healthy control group (10%). Among the Candida-positive patients, 16 of 35 (46%) carried C. krusei, while C. albicans was the second most common isolate (12 of 35 patients; 34%). The corresponding figures for the control group were 2 of 13 (15%) and 6 of 13 (46%), respectively. Studies of the antifungal resistance of the C. krusei isolates from patients indicated that all except one of the isolates were resistant to fluconazole, two isolates were resistant to ketoconazole, and all isolates were sensitive to amphotericin B. Evaluation of their genetic profiles by randomly amplified polymorphic DNA analysis with three different primers and subsequent analysis of the gel profiles by computerized cluster-derived dendrograms revealed that the C. krusei isolates from patients belonged to 10 disparate clusters, despite the origin from a single locale. These nascent findings indicate an alarmingly high prevalence of a Candida species resistant to a widely used antifungal in a part of the world where HIV disease is endemic.—Authors' Abstract

Shaw, I. N., Ebenezer, G., and Rao, G. S. Leprosy lesion on the prepuce of the male genitalia: a case report. Lepr. Rev. 73(3) (2002) 276–278.

A case of borderline leprosy in type I reaction with cutaneous lesions on the prepuce is reported. The need to examine the genitalia in all male leprosy patients is stressed.—Authors' Abstract

Zamir, E., Hudson, H., Ober, R. R., Kumar, S. K., Wang, R. C., Read RW, and Rao, N. A. Massive mycobacterial choroiditis during highly active antiretroviral therapy: another immune-recovery uveitis? Ophthalmology 109(11) (2002) 2144–2148.

PURPOSE: To describe the ocular presentation of disseminated mycobacterial disease occurring during immune-recovery in a patient with acquired immune deficiency syndrome (AIDS). STUDY DESIGN: Case report and literature review. PARTICIPANTS: A 41-year-old AIDS patient with a prior diagnosis of cytomegalovirus retinitis. METHODS: The patient developed progressive, bilateral multifocal choroiditis with panuveitis 2 months after beginning and responding to highly active antiretroviral therapy. His left eye became blind and painful and was enucleated. Pathologic examination revealed massive choroiditis with well-formed, discrete granulomas and multiple intracellular and extracellular acid-fast organisms within the choroidal granulomas. Culture and polymerase chain reaction of vitreous specimens revealed Mycobacterium avium complex (MAC). RESULTS: Empiric, and later sensitivity-guided, local and systemic antibiotic therapy was used to treat the remaining right eye, but it continued to deteriorate. Despite medical therapy, three vitrectomies and repeated intravitreal injections of amikacin, a total retinal detachment ensued. One week after the third vitrectomy, the patient died from mesenteric artery thrombosis in the setting of disseminated mycobacterial disease. CONCLUSIONS: This is the first report of ocular inflammation as the presenting finding in the recently recognized syndrome of immune-recovery MAC disease. Pathogenesis of this entity is related to an enhanced immune response to a prior, subclinical, disseminated infection. The formation of discrete granulomas, normally absent in MAC infections in AIDS, reflects this mechanism.—Authors' Abstract

Hostetter, J. M., Steadham, E. M., Haynes, J. S., Bailey, T. B., and Cheville, N. F. Cytokine effects on maturation of the phagosomes containing Mycobacteria avium subspecies paratuberculosis in J774 cells. FEMS Immunol. Med. Microbiol. 34(2) (2002) 127–134.

Mycobacterium avium subspecies paratuberculosis (M. a. ptb) is an intracellular pathogen of macrophages. Intracellular survival of several species of pathogenic mycobacteria is dependent on inhibition of maturation of the phagosomes containing these pathogens into functional phagolysosomes. In activated macrophages, however, this capacity is reduced, leading to increased bacterial killing. It is the hypothesis of this study that there is increased acidification and maturation of the phagosome containing M. a. ptb in interferon gamma and lipopolysaccharide (IFN-gamma/LPS) activated macrophages. In activated macrophages colocalization of M. a. ptb with either a marker of acidic compartments (Lysotracker Red) or compartments containing a late phagosome maturation marker lysosome-associated membrane protein-1 (Lamp-1) were evaluated by laser confocal microscopy. Intracellular survival of M. a. ptb in activated macrophages was evaluated directly using differential fluorescent live/dead staining. The results of this study demonstrated increased colocalization of both Lysotracker Red and Lamp-1 with FITC labeled M. a. ptb, which correlated with decreased survival of M. a. ptb within activated macrophages.—Authors' Abstract

Jacobs, M., Fick, L., Allie, N., Brown, N., and Ryffel, B. Enhanced immune response in Mycobacterium bovis Bacille Calmette Guerin (BCG)-infected IL-10-deficient mice. Clin. Chem. Lab. Med. 40(9) (2002) 893–902.

The role of the endogenous interleukin-10 (IL-10) in the control of Mycobacterium bovis Bacille Calmette Guerin (BCG) infection was assessed using IL-10-deficient (IL-10/) mice. Similar to wild-type (WT) mice, IL-10/ mice were resistant to intravenous challenge with Mycobacterium bovis BCG. Significantly higher plasma concentrations of IL-12 and tumour necrosis factor (TNF) indicated an elevated protective immune response of IL-10/ mice. Determination of bacilli burden in IL-10/ mice showed accelerated clearance in the lungs, spleen and the liver in comparison to WT mice. Enhanced inflammation and a vigorous granulomatous response accompanied accelerated mycobacterial clearance. Immunohistochemical analysis of hepatic granulomas from IL-10/ mice revealed augmented lymphocyte recruitment and macrophage activation, such as increased major histocompatibility complex (MHC) class II and inducible nitric oxide synthase (iNOS) expression. Further, it was found that enlarged granulomas persisted subsequent to mycobacterial clearance and failed to resolve in the absence of IL-10. In conclusion, endogenous IL-10 dampens the cell-mediated immune response to mycobacterial infection.—Authors' Abstract

Jason, J., Archibald, L. K., Nwanyanwu, O. C., Kazembe, P. N., Chatt, J. A., Norton, E., Dobbie, H., and Jarvis, W. R. Clinical and immune impact of Mycobacterium bovis BCG vaccination scarring. Infect. Immun. 70(11) (2002) 6188–6195.

The World Health Organization recommends Mycobacterium bovis BCG vaccination in areas of high tuberculosis prevalence. BCG's clinical and immune effects, not necessarily Mycobacterium tuberculosis specific, are unclear. BCG vaccine scarring often is used as a surrogate marker of vaccination or of effective vaccination. We evaluated BCG scarring status in relation to clinical findings and outcome in 700 hospitalized Malawians, of whom 32 had M. tuberculosis bloodstream infections (BSI) (10 of whom had cellular immune studies done) and of whom 48 were infants <6 months old and therefore recently vaccinated (19 of whom had immune studies). In the patients 6 months old, scarring was not related to the presence of pulmonary symptoms (35 versus 30%), chronic cough or fever, mortality, or M. tuberculosis BSI. In M. tuberculosis BSI patients, scarring was unrelated to mortality, vital signs, or clinical symptoms but those with scarring had higher proportions of memory and activated T cells and more type 2-skewed cytokine profiles. Infants with either BCG scarring (N = 10) or BCG lesional inflammation (N = 5) had no symptoms of sepsis, but 18 of 33 infants without BCG vaccination lesions did. Those with BCG lesions had localized infections more often than did those without BCG lesions. These infants also had lower median percentages of lymphocytes spontaneously making interleukin-4 (IL-4) or tumor necrosis factor alpha (TNF-alpha) and lower ratios of T cells spontaneously making IL-4 to T cells making IL-6. Thus, we found that, in older patients, BCG vaccine scarring was not associated with M. tuberculosis-specific or nonspecific clinical protection. Those with M. tuberculosis BSI and scarring had immune findings suggesting previous M. tuberculosis antigen exposure and induction of a type 2 cytokine pattern with acute reexposure. It is unlikely that this type 2 pattern would be protective against mycobacteria, which require a type 1 response for effective containment. In infants <6 months old, recent BCG vaccination was associated with a non-M. tuberculosis-specific, anti-inflammatory cytokine profile. That the vaccinated infants had a greater frequency of localized infections and lesser frequency of sepsis symptoms suggests that this postvaccination cytokine pattern may provide some non-M. tuberculosis-specific clinical benefits.—Authors' Abstract

Oriani, D. S. and Sagardoy, M. A. Nontuberculous mycobacteria in soils of La Pampa province (Argentina). Rev. Argent. Microbiol. 34(3) (2002) 132–137.

The presence of nontuberculous mycobacteria (NTM) was investigated in forty soil samples belonging to the four physiographic regions (Eastern, Central, Southern and Western) that constitute La Pampa province. The presence of NTM in 67.5% of these soil samples was determined. The density of mycobacteria ranged 25–4500 mycobacteria g¹ dry soil (mean = 516 CFU g¹). Significant differences were found in relation to both the investigated regions (p <0.01) and the soil pH (r = 0.44*) (p = 0.02). The mycobacteria represented less than 0.00001% of the total aerobic bacteria found in the soils. Twenty-seven isolated mycobacteria were classified according to the culture, biochemical, enzymatic characteristics and antibiotic sensitivity. Mycobacterium fortitium was the dominant mycobacterium and was detected in 63% of the positive soils. This species showed ability for living in sandy to sandy loam soils, within a wide pH range (6.5–9.7) and organic matter (4.15–83.63 g kg¹). Two other species were M. phlei (range = 50–4500 CFU g¹) and M. kansasii (range = 50–500 CFU g¹).—Authors' Abstract

Orrell, R. W., King, R. H., Bowler, J. V., and Ginsberg, L. Peripheral nerve granuloma in a patient with tuberculosis. J. Neurol. Neurosurg. Psychiatry 73(6) (2002)769–771.

The cause of peripheral neuropathy associated with tuberculosis is controversial. Possibilities include an immune mediated neuropathy, direct invasion of nerves, vasculitic neuropathy, compressive neuropathy, a meningitic reaction, and the toxic effects of antituberculous chemotherapy. This report describes the unusual finding of granulomas in the peripheral nerve of a patient with tuberculosis. The pathological findings were of a delayed hypersensitivity reaction, but with no more specific indications of the mechanism of the neuropathy.—Authors' Abstract

Sandor, M., Weinstock, J. V., and Wynn, T. A. Granulomas in schistosome and mycobacterial infections: a model of local immune responses. Trends Immunol. 24(1) (2003) 44–52.

Granulomatous immune responses are interesting models for the effector phase of immunity, in that granulomas can be part of both immune protection and disease pathology during the course of various infectious and autoimmune diseases. Focusing mainly on granulomas induced by Schistosoma or Mycobacterium infection, this article reviews T-cell recruitment, local cytokine networks and the regulation of cytokine networks by neuropeptides in granulomas. In addition, different activation pathways for macrophages residing in granulomas are discussed. Granulomas are unique inflammatory sites that offer a challenging and rewarding model to study immunity.—Authors' Abstract

Saunders, B. M., Dane, A., Briscoe, H., and Britton, W. J. Characterization of immune responses during infection with Mycobacterium avium strains 100, 101 and the recently sequenced 104. Immunol. Cell. Biol. 80(6) (2002) 544–549.

Mycobacterium avium strain 104 was chosen as the M. avium isolate to sequence, as it is virulent to humans, stable and readily transfectable. As this strain has not been widely studied we sought to investigate the pattern of 104 infection in mice. Bacterial growth and the immune response generated were compared with infection with the low virulence M. avium strain 100, and the high virulence common laboratory strain, 101. Mycobacterium avium strains 104 and 101 grew progressively within mice, while strain 100 was gradually cleared. Strains 104 and 101 induced strong T cell activation and spleen cell cultures produced similar levels of IFN-gamma. In mice infected with strain 100 no significant T cell activation or IFN-gamma production was measured. Further, mice infected with strain 104 or 101 also displayed comparable inflammatory responses and similar granuloma formation, while only minimal inflammation was in mice infected with strain 100. Strains 101 and 104 also grew in a similar fashion in bone-marrow-derived macrophages and induced significant levels of TNF and nitric oxide. Thus infection with M. avium strain 104 induced an immunological response comparable to M. avium strain 101 and, with the availability of its sequence, should be a useful tool for designing new vaccines or drugs therapies to treat the increasing incidence of M. avium infection in humans.—Authors' Abstract

Barbosa, A. A. Jr., Guimaraes, N. S., Follador, I., Sarno, L. S., and Pereira, C. P. Leprosy combined with elastolytic granuloma. An. Bras. Dermatol. 77(5) (2002) 585–592.

Two cases of leprosy combined with elastolytic giant cell granuloma are reported. Though a coincidental occurrence cannot be excluded, a possible pathogenic relationship between the two conditions is postulated. It is possible that an immunological mechanism plays a role in the elastolytic process, which could also be caused by actinic damage in the skin altered by leprosy.—An. Bras. de Dermatol.

Kang, T. J., Lee, S. B., and Chae, G. T. A polymorphism in the toll-like receptor 2 is associated with IL-12 production from monocyte in lepromatous leprosy. Cytokine 20(2) (2002) 56–62.

Toll-like receptor 2 (TLR2) is critical in the immune response to mycobacterial infections, and the mutations in the TLR2 have been shown to confer the susceptibility to infection with mycobacteria. We previously reported the detection of TLR2 Arg677Trp mutation in lepromatous leprosy. Here, the events triggered by TLR2 in response to cell lysate of Mycobacterium leprae (MLL), the causative agent of leprosy, were investigated. Upon stimulation with MLL, monocytes produced TNF-alpha and Interleukin-12 (IL-12), which play a role in the innate immune response to infection. Anti-TLR2 mAb blocked greater than 50% of the MLL-induced production of IL-12. We also performed the functional study on TLR2 by measurement of IL-12 production in serum and monocytes from leprosy patients with TLR2 mutation (Arg677Trp). The monocytes obtained from patients with the TLR2 mutation, in comparison to the wild-type TLR2, is significantly less responsive to MLL. It was also confirmed that patients with TLR2 mutation showed significantly lower serum levels of IL-12, in comparing with TLR2 wild-type. Our results reveal that innate immune response of monocytes against M. leprae is mediated by TLR2, and suggest that the mutation in the intracellular domain of TLR2 gene is associated with IL-12 production in lepromatous leprosy.—Authors' Abstract

Moudgil, K. D., Gupta, S. K., Naraynan, P. R., Srivastava, L. M., Mishra, R. S., and Talwar, G. P. Antibody response to phenolic glycolipid I and Mycobacterium w antigens and its relation to bacterial load in M. leprae-infected mice and leprosy patients. Clin. Exp. Immunol. 78(2) (1989) 214–218.

Twenty-six inbred BALB/cBy mice were infected with live Mycobacterium leprae by injecting 6 × 10(3) bacilli in the hind footpad. Bleeds were collected at monthly intervals. After 6 months, acid-fast bacilli (AFB) were harvested monthly from the footpad of mice. The sera were analysed in enzyme immunoassay for antibodies against phenolic glycolipid I (PGL-I) of M. leprae and antigens of Mycobacterium w (M. w); 21 out of 26 (80.7%) mice demonstrated the presence of antibodies against PGL-I and M. w. Anti-M. w antibodies appeared slightly earlier than did anti-PGL-I antibodies. The titre of anti-M. w antibodies was higher than that of anti-PGL-I antibodies. The mice giving a positive antibody response had more than 7 × 10(5) AFB/footpad. The coefficient of correlation (r) between the number of AFB and antibody titres at the time of harvest was 0.566 for PGL-I and 0.628 for M. w. The value of r for bacterial index and antibody titres in 188 leprosy patients was 0.510 for PGL-I and 0.418 for M. w; these values were statistically significant (p <0.001). The decrease in bacterial index and antibody titres in treated lepromatous leprosy patients correlated with increase in the duration of chemotherapy. The measurement of anti-PGL-I antibodies of IgM class may serve as an adjunct to skin biopsy and skin-slit smear for serial monitoring of the bacterial load in the course of chemotherapy in leprosy control programs.—Authors' Abstract

Santos, A. R., Suffys, P. N., Vanderborght, P. R., Moraes, M. O., Vieira, L. M., Cabello, P. H., Bakker, A. M., Matos, H. J., Huizinga, T. W., Ottenhoff, T. H., Sampaio, E. P., and Sarno, E. N. Role of tumor necrosis factor-alpha and interleukin-10 promoter gene polymorphisms in leprosy. J. Infect. Dis. 186(11) (2002) 1687–1691.

Single-nucleotide polymorphisms within the genes coding for tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 have been associated with several infectious diseases. To determine whether such polymorphisms are associated with leprosy, genotyping was performed at the 308 and 238 positions of the promoter of the TNF-alpha gene in 210 and 191 patients with multibacillary (MB) leprosy, respectively; 90 and 79 patients with paucibacillary; and 92 control subjects. for the 592 and 819 positions within the promoter of the IL-10 gene, 143 patients with MB leprosy, 79 patients with PB leprosy, and 62 control subjects were included in the analysis. TNF2 allele frequency was significantly higher among control subjects than among all aptients with leprosy or in the MB group (p <.05 and p <.01). For the IL-10 gene, the frequency of the homozygous 819TT genotype was significantly higher among patients than among control subjects. These data indicate that a relationship exists between TNF-alpha and IL-10 promoter polymorphisms and the development of PB leprosy.—Authors' Abstract

Teles, R. M., Moraes, M. O., Geraldo, N. T., Salles, A. M., Sarno, E. N., and Sampaio, E. P. Differential TNFalpha mRNA regulation detected in the epidermis of leprosy patients. Arch. Dermatol. Res. 294(8) (2002) 355–362.

The epidermis is an important site of the immunoinflammatory response in the skin. In the present study, the expression of cytokine and ICAM-1 (intercellular adhesion molecule-1) genes was evaluateed by RT-PCR in the epidermis isolated from biopsies from 25 reactional leprosy patients. TNFalpha and IL-6 mRNAs were detected in all individuals during the reactional state (reversal reaction or erythema nodosum leprosum), IL-8 message was detected in 66.6% and 62.5% of the patients, IL-12 mRNA was present in 91.6% and 62.5% and ICAM-1 in 100% and 71.4%, respectively. In addition, when skin biopsies were obtained from the same patients before and during the reactional episode, an enhancement in cytokine mRNA, but not in ICAM-1 mRNA, was observed. Seven patients were also evaluated at the onset of reaction and during antiinflammatory treatment. In contrast to a preferential disease in the TNFalpha gene detected in the dermis, during the treatment phase, persistent/enhanced TNFalpha mRNA expression was detected in the epidermis in six out of the seven patients assessed. This peculiar pattern of expression might reflect a differential impact that in vivo antiinflammatory therapy has on the epidermis. The present findings indicate that the epidermis plays an important role in the local inflammatory response in leprosy and that the profile of response detected in the epidermis during the reactions may be regulated differently from that in the dermis.—Authors' Abstract

Arend, S. M., Van Meijgaarden, K. E., De Boer, K., De Palou, E. C., Van Soolingen, D., Ottenhoff, T. H., and Van Dissel, J. T. Tuberculin skin testing and in vitro T-cell responses to ESAT-6 and culture filtrate protein 10 after infection with Mycobacterium marinum or Mycobacterium kansasii. J. Infect. Dis. 186(12) (2002) 1797–1807.

T cell responses to ESAT-6 and culture filtrate protein 10 (CFP-10), antigens expressed by Mycobacterium tuberculosis but not by M. bovis bacille Calmette-Guerin (BCG), were found to discriminate reliably between infection with M. tuberculosis and BCG vaccination. Because the esat-6 and cfp-10 genes occur in M. kansasii and M. marinum, T cell responses to ESAT-6 and CFP-10 were investigated in patients infected with M. kansasii or M. marinum, persons intensively exposed to environmental mycobacteria, and unexposed control subjects. Tuberculin skin tests were performed, and peripheral blood mononuclear cells were cocultured with ESAT-6, CFP-10, peptide mixtures of ESAT-6 and CFP-10, and control antigens. When enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot assay (ELISPOT) were used to measure interferon-gamma production, most M. kansasii- or M. marinum-infected patients and several persons exposed to environmental mycobacteria were found to respond to ESAT-6 and/or CFP-10. ELISA and ELISPOT yielded comparable results, as did whole antigen and peptides (p <.0001). These results may be relevant for the development of novel assays for diagnosis of tuberculosis.—Authors' Abstract

Biet, F., Kremer, L., Wolowczuk, I., Delacre, M., and Locht, C. Mycobacterium bovis BCG producing interleukin-18 increases antigen-specific gamma interferon production in mice. Infect. Immun. 70(12) (2002) 6549–6557.

Interleukin-18 (IL-18) and IL-12 play a critical role in the expression of cell-mediated immunity involved in host defense against intracellular pathogens. Both cytokines are produced by macrophages and act in synergy to induce gamma interferon (IFN-gamma) production by T, B, and natural killer cells. In the present study, we analyzed both cellular and humoral responses upon infection with IL-18-secreting BCG of BALB/c and C3H/HeJ mice, two strains known to differ in their ability to support the growth of BCG. The cDNA encoding mature IL-18 was fused in frame with the alpha-antigen signal peptide-coding sequence, cloned downstream of the mycobacterial hsp60 promoter and expressed in BCG. IL-18 produced by the recombinant BCG strain was functional, as judged by NF-kappaB-mediated luciferase induction in a tissue culture assay. When susceptible mice were infected with IL-18-producing BCG, their splenocytes were found to produce higher amounts of Th1 cytokines after stimulation with mycobacterial antigens than the splenocytes of mice infected with the nonrecombinant BCG. This was most prominent for IFN-gamma, although the mycobacterial antigen-specific secretion of granulocyte-macrophage colony-stimulating factor and IL-10 was also augmented after infection with the recombinant BCG compared to infection with nonrecombinant BCG. In contrast, the immunoglobulin G levels in serum against mycobacterial antigens were lower when the mice were infected with IL-18-producing BCG compared to infection with nonrecombinant BCG. The IL-18 effect was delayed in BALB/c compared to C3H/HeJ mice. These results indicate that the production of IL-18 by recombinant BCG may enhance the immunomodulatory properties of BCG further toward a Th1 profile. This may be particularly useful for immunotherapeutic or prophylactic interventions in which a Th1 response is most desirable.—Authors' Abstract

Cardoso, F. L., Antas, P. R., Milagres, A. S., Geluk, A., Franken, K. L., Oliveira, E. B., Teixeira, H. C., Nogueira, S. A., Sarno, E. N., Klatser, P., Ottenhoff, T. H., and Sampaio, E. P. T-cell responses to the Mycobacterium tuberculosis-specific antigen ESAT-6 in Brazilian tuberculosis patients. Infect. Immun. 70(12) (2002) 6707–6714.

The Mycobacterium tuberculosis-specific ESAT-6 antigen induces highly potent T-cell responses and production of gamma interferon (IFN-gamma), which play a critical role in protective cell-mediated immunity against tuberculosis (TB). In the present study, IFN-gamma secretion by peripheral blood mononuclear cells (PBMCs) in response to M. tuberculosis ESAT-6 in Brazilian TB patients was investigated in relation to clinical disease types, such as pleurisy and cavitary pulmonary TB. Leprosy patients, patients with pulmonary diseases other than TB, and healthy donors were assayed as control groups. Sixty percent of the TB patients indeed recognized M. tuberculosis ESAT-6, as did 50% of the leprosy patients and 60% of the non-TB controls. Nevertheless, the levels of IFN-gamma in response to the antigen ESAT, but not to antigen 85B (Ag85B) and purified protein derivative (PPD), were significantly lower in controls than in patients with treated TB or pleural or cavitary TB. Moreover, according to Mycobacterium bovis BCG vaccination status, only 59% of the vaccinated TB patients responded to ESAT in vitro, whereas 100% of them responded to PPD. Both CD4 and CD8 T cells were able to release IFN-gamma in response to ESAT. The present data demonstrate the specificity of ESAT-6 of M. tuberculosis and its ability to discriminate TB patients from controls, including leprosy patients. However, to obtain specificity, it is necessary to include quantitative IFN-gamma production in response to the antigen as well, and this might limit the use of ESAT-6-based immunodiagnosis of M. tuberculosis infection in an area of TB endemicity.—Authors' Abstract

Chackerian, A., Alt, J., Perera, V., and Behar, S. M. Activation of NKT cells protects mice from tuberculosis. Infect. Immun. 70(11) (2002) 6302–6309.

The T-cell immune response to Mycobacterium tuberculosis is critical in preventing clinical disease. While it is generally accepted that both major histocompatibility complex class I (MHC-I)-restricted CD8(+) and MHC-II-restricted CD4(+) T cells are important for the immune response to M. tuberculosis, the role of non-MHC-restricted T cells is still not clearly delineated. We have previously reported that CD1d(/) mice do not differ from CD1d(+/+) mice in their survival following infection with M. tuberculosis. We now show that, although CD1d-restricted NKT cells are not required for optimum immunity to M. tuberculosis, specific activation of NKT cells by the CD1d ligand alpha-galactosylceramide protects susceptible mice from tuberculosis. Treatment with alpha-galactosylceramide reduced the bacterial burden in the lungs, diminished tissue injury, and prolonged survival of mice following inoculation with virulent M. tuberculosis. The capacity of activated NKT cells to stimulate innate immunity and modulate the adaptive immune response to promote a potent antimicrobial immune response suggests that alpha-galactosylceramide administration could have a role in new strategies for the therapy of infectious diseases.—Authors' Abstract

Ciaramella, A., Cavone, A., Santucci, M. B., Amicosante, M., Martino, A., Auricchio, G., Pucillo, L. P., Colizzi, V., and Fraziano, M. Proinflammatory cytokines in the course of Mycobacterium tuberculosis-induced apoptosis in monocytes/macrophages. J. Infect. Dis. 186(9) (2002) 1277–1282.

Mycobacterium tuberculosis (MTB) can induce apoptosis in monocytes/macrophages both in vitro and in vivo, and this phenomenon is associated with mycobacterial survival. The present study addresses the possibility that apoptotic and inflammatory pathways could coexist through a caspase-1-mediated mechanism. In this context, a caspase-1 inhibitor (YVAD), but not caspase-3 (DEVD) or caspase-4 (LEVD) inhibitors, was able to strongly inhibit MTB-induced apoptosis. Moreover, caspase-1 activity was confirmed by the increased maturation of interleukin (IL)-1beta. Of interest, IL-1beta and tumor necrosis factor (TNF)-alpha were produced massively in the course of infection, and both were inhibited by YVAD pretreatment. To determine whether TNF-alpha was produced actively by apoptotic cells, the intracytoplasmatic cytokine content and apoptotic phenotype were analyzed at the single-cell level. Results showed a progressive increase of TNF-alpha production in annexin V-positive cells. These results indicate that MTB-induced apoptosis is associated with proinflammatory cytokine production.—Authors' Abstract

Davis, J. M., Clay, H., Lewis, J. L., Ghori, N., Herbomel, P., and Ramakrishnan, L. Real-time visualization of mycobacterium-macrophage interactions leading to initiation of granuloma formation in zebrafish embryos. Immunity 17(6) (2002) 693–702.

Infection of vertebrate hosts with pathogenic Mycobacteria, the agents of tuberculosis, produces granulomas, highly organized structures containing differentiated macrophages and lymphocytes, that sequester the pathogen. Adult zebrafish are naturally susceptible to tuberculosis caused by Mycobacterium marinum. Here, we exploit the optical transparency of zebrafish embryos to image the events of M. marinum infection in vivo. Despite the fact that the embryos do not yet have lymphocytes, infection leads to the formation of macrophage aggregates with pathological hallmarks of granulomas and activation of previously identified granuloma-specific Mycobacterium genes. Thus, Mycobacterium-macrophage interactions can initiate granuloma formation solely in the context of innate immunity. Strikingly, infection can redirect normal embryonic macrophage migration, even recruiting macrophages seemingly committed to their developmentally dictated tissue sites.—Authors' Abstract

Dieli, F., Ivanyi, J., Marsh, P., Williams, A., Naylor, I., Sireci, G., Caccamo, N., Di Sano, C., and Salerno, A. Characterization of lung gammadelta T-cells following intranasal infection with Mycobacterium bovis Bacillus Calmette-Guerin. J. Immunol. 170(1) (2003) 463–469.

The lungs are considered to have an impaired capacity to contain infection by pathogenic mycobacteria, even in the presence of effective systemic immunity. In an attempt to understand the underlying cellular mechanisms, we characterized the gammadelta T cell population following intranasal infection with Mycobacterium bovis bacillus Calmette-Guerin (BCG). The peak of gammadelta T cell expansion at 7 days postinfection preceded the 30 day peak of alphabeta T cell expansion and bacterial count. The expanded population of gammadelta T cells in the lungs of BCG-infected mice represents an expansion of the resident Vgamma2 T cell subset as well as an influx of Vgamma1 and of four different Vdelta gene-bearing T cell subsets. The gammadelta T cells in the lungs of BCG-infected mice secreted IFN-gamma following in vitro stimulation with ionomycin and PMA and were cytotoxic against BCG-infected peritoneal macrophages as well as against the uninfected J774 macrophage cell line. The cytotoxicity was selectively blocked by anti-gammadelta TCR mAb and strontium ions, suggesting a granule-exocytosis killing pathway. Depletion of gammadelta T cells by injection of specific mAb had no effect on the subsequent developing CD4 T cell response in the lungs of BCG-infected mice, but significantly reduced cytotoxic activity and IFN-gamma production by lung CD8 T cells. Thus, gammadelta T cells in the lungs might help to control mycobacterial infection in the period between innate and classical adaptive immunity and may also play an important regulatory role in the subsequent onset of alphabeta T lymphocytes.—Authors' Abstract

Dieli, F., Sireci, G., Caccamo, N., Di Sano, C., Titone, L., Romano, A., Di Carlo, P., Barera, A., Accardo-Palumbo, A., Krensky, A. M., and Salerno, A. Selective depression of interferon-gamma and granulysin production with increase of proliferative response by Vgamma9/Vdelta2 T-cells in children with tuberculosis. J. Infect. Dis. 186(12) (2002) 1835–1839.

Vgamma9/Vdelta2 T cells can contribute to protective immune response against Mycobacterium tuberculosis, although the extent to which and mechanisms by which they could actually protect against human tuberculosis remain unclear. We have previously reported that Vgamma9/Vdelta2 T cells from tuberculin purified protein derivative (PPD)-positive children, either healthy or affected by different clinical forms of tuberculosis, strongly proliferate to different phosphoantigens in vitro, whereas Vgamma9/Vdelta2 T cells from PPD-negative healthy subjects proliferate very poorly. We report here that Vgamma9/Vdelta2 T cells from tuberculous children have an increased proliferative activity, but decreased interferon (IFN)-gamma production and granulysin expression. After successful chemotherapy, the Vgamma9/Vdelta2 T cell proliferative response strongly decreased, whereas IFN-gamma and granulysin production consistently increased. Disease-associated changes in Vgamma9/Vdelta2 T cell effector functions in patients with tuberculosis are consistent with the possibility that these T cells may play a protective role in immune response against M. tuberculosis infection.—Authors' Abstract

D-Souza, S., Rosseels, V., Romano, M., Tanghe, A., Denis, O., Jurion, F., Castiglione, N., Vanonckelen, A., Palfliet, K., and Huygen, K. Mapping of murine Th1 helper T-cell epitopes of mycolyl transferases Ag85A, Ag85B, Ag85C from Mycobacterium tuberculosis. Infect. Immun. 71(1) (2003) 483–493.

BALB/c (H-2(d)) and C57BL/6 (H-2(b)) mice were infected intravenously with Mycobacterium tuberculosis H37Rv or vaccinated intramuscularly with plasmid DNA encoding each of the three mycolyl transferases Ag85A, Ag85B, and Ag85C from M. tuberculosis. Th1-type spleen cell cytokine secretion of interleukin-2 (IL-2) and gamma interferon (IFN-gamma) was analyzed in response to purified Ag85 components and synthetic overlapping peptides covering the three mature sequences. Tuberculosis-infected C57BL/6 mice reacted strongly to some peptides from Ag85A and Ag85B but not from Ag85C, whereas tuberculosis-infected BALB/c mice reacted only to peptides from Ag85A. In contrast, spleen cells from both mouse strains produced elevated levels of IL-2 and IFN-gamma following vaccination with Ag85A, Ag85B, and Ag85C DNA in response to peptides of the three Ag85 proteins, and the epitope repertoire was broader than in infected mice. Despite pronounced sequence homology, a number of immunodominant regions contained component specific epitopes. Thus, BALB/c mice vaccinated with all three Ag85 genes react