Bacillus

Surv Ophthalmol, 2002 May-Jun, 47(3), 275 - 87
The ophthalmic trials of G . H . A . Hansen; Marmor MF; G . H . A . Hansen (1841-1912) is widely known as the discoverer of the infectious cause of leprosy . It is less well known that his career was threatened by an episode involving experimentation on the eye . As a staff physician at the leprosy hospitals of Bergen, Norway, early in his career, Hansen learned about ocular involvement in leprosy and co-authored Leprous Diseases and the Eye . In 1873 he observed bacilli in leprous nodules, but proof of an infectious origin was difficult to obtain because the agent could not be cultured and no one had demonstrated direct transmission . Hansen tried several unsuccessful experiments, and in 1879 he passed a cataract knife that had incised an active leprous nodule into a woman's conjunctiva . No nodule developed, but the woman complained of pain and said she was never asked for permission . Hansen was brought to trial where eminent physicians testified on his behalf-but Hansen himself readily admitted that no permission had been sought for fear the woman would say no . He was convicted, and relieved of his post as staff physician, but he was allowed to retain an appointment as Chief Medical Officer of Health for Leprosy, in which capacity he worked for the rest of his life.

J Am Vet Med Assoc, 2002 Jun 1, 220(11), 1661 - 3, 1650
Pulmonary mycobacteriosis caused by Mycobacterium haemophilum and M . marinum in a royal python; Hernandez-Divers SJ et al.; An adult female royal python was referred with an 18-month history of chronic respiratory tract disease . Anemia and moderate leukocytosis with heterophilia and monocytosis were detected and interpreted as evidence of a chronic inflammatory condition . Evaluation of lateral and dorsoventral radiographic views revealed multiple soft-tissue opacities within the cranial lung fields . Endoscopic evaluation revealed that the normal reticulated pattern on the surface of the lung had been largely replaced by diffuse, granulomatous tissue . Histologic examination of biopsy specimens revealed classic pyogranulomas . Ziehl-Neelsen stains revealed numerous acid-fast bacilli consistent with Mycobacterium spp . Molecular methods including polymerase chain reaction restriction assays and DNA sequencing confirmed the identification of M . haemophilum and M . marinum . The snake was euthanatized . Mycobacteriosis is an uncommon and sporadic pyogranulomatous disease of reptiles . In most cases of reptile mycobacteriosis, treatment is not advised because of the chronic nature and often advanced stage of the disease, long-term and expensive nature of potential treatment regimens, and the risk of spread to other animals, including humans.

J Forensic Sci, 2002 May, 47(3), 640 - 4
Fatal meningitis and encephalitis due to Bartonella henselae bacteria; Gerber JE et al.; Bacterial infection due to Bartonella henselae commonly develops in children and young adults following cat/dog contacts and/or cat/dog scratches . Regional lymphadenopathy is its most common clinical expression . However, encephalitis and Parinaud's syndrome (oculoglandular syndrome) have also been reported as has systemic illness . A review of the international literature in all languages revealed no fatal complications in immunocompetent hosts . A four-year-old white child with no underlying illness began to have seizure-like activity . She was taken to a local hospital and subsequently transferred to a medical center . The child was treated aggressively for seizures and fever of unknown origin . However, her condition rapidly declined and she died without a specific diagnosis . At autopsy there was marked cerebral edema with no gross evidence of acute meningitis . Microscopic exams revealed multiple granulomatous lesions as well as a meningitis and encephalitis . A variety of cultures and stains were negative for acid fast and fungal organisms . Warthin-Starry stains of involved tissue including brain and liver revealed pleomorphic rod shaped bacilli consistent with Barronella henselae . Analysis of brain tissue with polymerase chain reaction (PCR) and Southern blot for the deoxyribonucleic acid (DNA) was definitive for DNA of Bartonella henselae bacteria.

Acta Chir Belg, 2002 Apr, 102(2), 118 - 21
Tuberculous lymphadenopathy in adults: a review of 35 cases; Mert A et al.; We retrospectively reviewed clinical, diagnostic, therapeutic and prognostic features of 35 patients (25 female, 10 male, mean age: 33 years, range: 16-70) with tuberculous lymphadenopathy (TB LAP) which had been followed since 1980 . The diagnosis was established by tissue sampling in 32 cases (caseating granulomatous adenitis in 89%) or presence of acid-fast bacilli (AFB) in the aspirate in 2 cases and in the drainage in 1 case . Paraffin-embedded granulomatous tissues were stained by Ehrlich-Ziehl-Neelsen (EZN) and also Mycobacterium tuberculosis DNA was studied by polymerase chain reaction (PCR) (n = 21) . The patients were admitted with enlarging LAP (34%), draining LAP (9%), and both systemic complaints and enlarging LAP (57%) . Cervical lymph nodes were the most frequently involved site (77%) . Pathologic findings on chest X-ray were seen in 23% . Erythrocyte sedimentation rate (ESR) was higher than 100 mm/hour in 25% and associated with systemic complaints . Tuberculin skin test was positive in 91% . AFB could not be seen in any granulomatous tissue (n: 21), but PCR study was positive in 33% (7/21) . All patients were given anti-TB treatment (INH,RMP,EMB and/or PZA) . Surgical excision of draining LAP with surrounding inflammatory tissues in addition to the medical treatment was needed in 2 cases . Clinical improvement was obtained within 3 months of the treatment and ESR returned to normal within 5 months . After completion of the treatment, 22 patients were followed-up; mean duration was 3 years, and none relapsed . In conclusion, a patient with TB LAP generally presents with a few small, painless, cervical lymph nodes, which are slowly enlarging . For exact diagnosis, excisional biopsy for histologic and microbiologic studies is essential . Use of anti-TB drugs is the main therapeutic option.

Orbit, 2000 Mar, 19(1), 41 - 44
Orbital cellulitis with gas; Cheema RA et al.; Orbital cellulitis due to presumed gas-producing organisms has been associated with poor visual prognosis . We report successful visual recovery in a 14-year-old boy with orbital cellulitis and gas in the orbit on CT scan . The patient was treated with intravenous antibiotics and prompt orbital decompression . Pus from the sinus aspirate and orbital drain grew H . influenzae and Gram-negative bacilli . Repeat orbital surgery with drainage of residual abscess was performed and the patient made a complete recovery .

J Biotechnol, 2002 Jul 3, 96(3), 259 - 70
Cultivation of Mycobacterium bovis BCG in bioreactors; Dietrich G et al.; The Mycobacterium bovis BCG vaccine for commercial use is classically produced as surface pellicles by culture on synthetic medium . Under these conditions, reproducibility of the cultures and quality assessment are hampered by slow growth of the bacilli, the formation of bacterial aggregates and a high proportion of dead bacilli after processing and final formulation of the vaccine . Here, we established dispersed cultures of M . bovis BCG in synthetic media in small-scale bioreactors . These cultures allow recording and adjusting of culture parameters and give rise to single bacilli with a high degree of live bacteria . In the murine model, bioreactor-grown M . bovis BCG exhibited slightly stronger replication and persistence than the vaccine produced under the classical conditions . The protective efficacy against challenge with M . tuberculosis was identical for both vaccine preparations.

Rev Med Chil, 2002 Mar, 130(3), 319 - 21
{Tuberculosis bursitis: report of case}; Perez C et al.; We report a previously healthy 51 years old woman with a one year history of pain in the left hip associated with a mass without fever or local inflammatory changes . Magnetic resonance imaging located the mass in the medial gluteal muscle . The pathological examination of the mass disclosed unspecific inflammatory changes . Due to worsening of pain, a left throchanteric bursitis was diagnosed two weeks later and a throchanteric bursectomy was performed, obtaining a second biopsy . Aerobic, anaerobic and fungal cultures were negative . Lowenstein-Jensen culture showed development of mycobacteria and a genetic probe confirmed the presence of Mycobacterium tuberculosis . The second biopsy also identified several granulomas with areas of caseation and the Ziehl-Nielsen stain was positive for acid fast bacilli . Osteoarticular tuberculosis, specially bursitis, should be suspected in the presence of chronic pain associated with swelling.

Southeast Asian J Trop Med Public Health, 2001 Dec, 32(4), 835 - 43
Simple microplate hybridization assay for detection of amplified products of Mycobacterium tuberculosis; Tansuphasiri U et al.; We describe a simple microplate hybridization assay for the rapid detection of the IS6110 PCR products of Mycobacterium tuberculosis from clinical cultures and from sputum specimens . The assay is based on the specific detection with a fluorescein-labeled detection probe of biotinylated PCR products which are captured on avidin coated microplate . Hybridized products with fluorescein were identified by using anti-fluorescein antibody, horseradish peroxidase conjugate and colorimetric peroxidase substrate . The specificity of the assay was assessed by analysis of 56 bacterial strains: the assay discriminated perfectly between the positive and negative groups when an OD490 of 0.18 was used as the cut-off point . The assay was sensitive enough to detect as little as 1 pg of M . tuberculosis H37Rv DNA, which is equivalent to approximately three bacilli . To evaluate the assay performance clinically, 190 sputum samples from newly diagnosed TB patients were tested; 79 were classified as TB positive, and 111 were classified as TB negative by culture and acid-fast staining as the 'gold standard' . The sensitivity, specificity and accuracy of the PCR-microplate hybridization assay were 90, 100 and 96%, respectively . The total assay time of hybridization following the PCR was 4 hours . The PCR-microplate hybridization assay is fast, simple and accurate and is suitable for use in the microbiology laboratory or for the analysis of large numbers of samples.

Rev Mal Respir, 2002 Apr, 19(2 Pt1), 161 - 5
{Tuberculosis of the lung bases}; Ben Miled-M'rad K et al.; Tuberculosis of the lung bases is a rare condition that can mimic pneumonia, bronchial carcinoma, lung abscess or bronchiectasis . Diagnostic delays, that can lead to serious complications not amenable to surgical resection, are often the result of repeatedly negative bacteriological examinations . We report a series of 10 cases of tuberculosis of the lung bases collected between 1993 and 1998 that were examined by computerised tomography . The chest x-ray most commonly showed a heterogeneous opacity . On the CT scan this corresponded with an area of alveolar consolidation which, when it was associated with centrilobular micronodules and cavitated nodules and masses, suggested the diagnosis of basal tuberculosis . Indeed the absence of apical lesions and the frequency of involvement of mediastinal and bronchial lymph nodes support the hypothesis of bronchial dissemination of the bacilli from tuberculous nodes . This explanation is supported by the high incidence of endobronchial lesions found at bronchoscopy in this form of tuberculosis.

Appl Environ Microbiol, 2002 Jun, 68(6), 3159 - 61
Identification of nontuberculous mycobacteria existing in tap water by PCR-restriction fragment length polymorphism; Chang CT et al.; This paper presents the finding of the possible cause of the high false-positive rate in acid-fast staining in histological examinations . Using acid-fast staining, culture, and PCR, acid-fast bacilli were detected in 83.7% of 49 hospital tap water samples and nontuberculous mycobacteria (NTM) were detected in 20.4% of the same 49 samples . The 10 NTM isolates were also identified to the species level using PCR-restriction fragment length polymorphism . Our findings indicate that NTM in hospital tap water are the possible cause of false positives in acid-fast staining and of nosocomial infection in immunocompromised patients.

Ann Med Interne (Paris), 2002 Mar, 153(2), 75 - 81
{Neuromeningeal tuberculosis in northeastern suburbs of Paris . Nineteen cases}; Grimaud-Ayina M et al.; SUBJECT: Analyze characteristics of neuromeningeal tuberculosis in the northeastern suburbs of Paris . MATERIAL AND METHOD: Retrospective study of 19 observations between 1988 and 1999 . RESULTS: Thirteen cases of meningitis, 3 cases of meningitis associated with a tuberculoma and 3 cases with isolated tuberculoma were described . Young subjects (average age 46.4), foreign born (73.7%) were preferentially affected . Three patients were infected by the human immunodeficiency virus . Weight-loss and weakness (84%), fever (79%), headache (63%) dominate the clinical manifestations . The diagnosis delay was 56.6 days, and the therapeutic time, 8.6 days . The search for acid fast bacilli was positive only in one case on direct examination of the cerebrospinal fluid, whereas the culture was positive in 68.7% of the cases . Cerebral imagery was abnormal in13 patients . Average duration of antituberculous treatment was 13,4 months (range: 9-24) . Corticosteroids were associated in 12 patients . Thirteen patients were cured, 3 died and 3 were lost to follow up . CONCLUSION: The characteristics of neuromeningeal tuberculosis are not modified, but the diagnosis delay of the neuromeningeal tuberculosis, which conditions the prognosis, remains significant . The optimal duration of the antituberculous treatment and the conditions of the steroid therapy have to be specified.

Life Sci Space Res, 1965, 3, 142 - 8
Ultra-high vacuum and microorganisms; Imshenetsky AA et al.; Experimental study of the space environment effect on living organisms is one of the most important tasks of exobiology . However location of the living organisms under study outside space vehicles and sputniks is combined with technical difficulties . Therefore it is much more suitable to reproduce conditions found in space in laboratory and to study its effect on living cells . It was proved that low temperatures, even those approaching absolute zero do not kill microorganisms . The fact of microorganisms surviving radiation of some million roentgen suggests that they would not be killed by ionizing radiation in space . UV rays should be considered as the most active sterilizing agent . The ability of microorganisms to resist high vacuum is studied much worse . Under the action of high vacuum at 10(-8)-10(-9) mm Hg during 72 hours all studied seven species of spore-forming bacilli remained viable . As for nonsporeforming bacteria under conditions mentioned, cells of some species perished (correction of parished) completely while other species retained viable cells . Conidia of fungi and parts of mycelium of fungi which do not form conidia, sustained high vacuum well.

Indian J Pathol Microbiol, 2001 Oct, 44(4), 435 - 8
Prevalence of gram negative anaerobic bacilli in routine clinical specimens; De A et al.; A total of 206 gram negative anaerobic bacilli were isolated from 2591 routine clinical specimens . Majority of them (78.2%) were part of a mixed flora . A high incidence of gram negative anaerobic bacilli was found in post-operative wound infections and diabetic foot infections (30.1%), followed by necrotizing fasciitis and crush injuries (18%), pus of empyema and lung abscesses (14.5%) and aspirates from other deep abscesses (10.7%) . Bacteroides fragilis was the most frequently recognized species (42.2%), followed by Fusobacterium sp . (17.4%), Prevotella sp . (15.1%) and Porphyromonas sp . (11.2%) . Neomycin blood agar plate yielded the maximum number (72.8%) of strains in pure culture . Gram negative anaerobic bacilli in infective processes are significantly on the rise and so suspected cultures should be processed for both aerobic and anaerobic organisms.

Kekkaku, 2002 Apr, 77(4), 361 - 6
{A case of multiple bone and joint tuberculosis which had been misdiagnosed as the rheumatoid arthritis and treated with prednisolone for eleven months}; Tsuduki E et al.; A 34-year-old man had a multiple arthralgia for about eleven months . The swelling of his right wrist and foot had appeared in the dorsal side, and he had been misdiagnosed as the rheumatoid arthritis . He was treated with prednisolone in the dosages of 2.5 mg per day for one month, and 10 mg per day for ten months . When he admitted to our hospital, the bone X-ray examinations of the wrist and foot revealed the marked atrophy and destruction of the carpal and tarsal bones . The aspiration fluid from the swelling around his wrist and foot was positive for acid-fast bacilli on smear and Mycobacterium tuberculosis was found on culture . He was treated with isoniazid, rifampicin, ethambutol and pyrazinamide, however, these medication was not adequately effective to his complications of tuberculous arthritis . Curettage, irrigation and synovectomy of his right carpal and tarsal bone were performed in order to control his bone and joint infection . He recovered from his arthritis and tenosynovitis after these operations . The clinical practitioners should not omit tuberculosis from the differential diagnosis of persistent osteoarthralgia.

Kekkaku, 2002 Apr, 77(4), 329 - 39
{Discussing the current situation of tuberculosis case-finding by mass miniature radiography in Japan}; Ohmori M et al.; The system of tuberculosis (TB) case-finding by mass miniature radiography (MMR) was established and expanded for almost all Japanese citizens in the 1950s . And, as stipulated by the TB Prevention Law, periodic mass screenings for schools, inhabitants, employees and institutions have been carried out . Among those aged over 25 years, the proportion of people screened by MMR was estimated to be 60.3% . This means that about 54 million people aged over 25 years are receiving medical service with MMR every year . However, the detection rates of TB cases by MMR have declined markedly compared with those in 1950s . As of 1998, the detection rate was 0.03 per 1,000 for school children and students, 0.06 per 1,000 for employees, and 0.16 per 1,000 for inhabitants . The proportion of cases detected by MMR among newly notified TB cases was 12.8% in 1998, and this ratio has been almost constant for the last 10 years . This ratio was greater among young adult TB cases . Approximately 20% of notified TB cases aged 20-39 years were detected by MMR for employees . Although the purpose of MMR is to find the cases before discharging TB bacilli, 35.1% of the cases were bacteriologically confirmed, and this proportion was greater among elderly TB cases . The Japan Anti-Tuberculosis Association (JATA) has been carrying out MMR for a long time . Eight selected branches of JATA that has been doing high quality case-finding reported 228 TB cases out of 965,440 inhabitants aged over 40 years examined by MMR in 1996 . Based on these results, the cost per TB case detected by MMR was calculated . The cost was 4.4 millions yen (yen) per case for all forms of TB, yen 2.3 millions for male, yen 8.4 millions for female, yen 7.3 millions for those aged 40-49 years and yen 1.8 millions for those aged over 80 years . TB detection rate by MMR for inhabitants was correlated with TB incidence rate in various areas, and based on this correlation, the cost was calculated for various incidence rates . For all forms of TB, the cost was yen 4.0 millions per case for an incidence rate of 30 per 100,000, and yen 6.7 millions for an incidence rate of 20 per 100,000 . MMR is not economically cost-effective even among elderly people and in areas with incidence rate less than 50 per 100,000, because the medical expense for a TB patient treated under hospitalization for 2 months and outpatient's clinic for 4 months is approximately yen 0.9 millions in 1996 . The decision making in continuation or abolition or limitation of MMR should be discussed from a wide range of cost-effectiveness analyses as well as from the view of public health service and willingness of people . For the purpose of decision making, this study provides the detection rates; the costs stratified by sex, age and incidence; and the proportion of cases detected by MMR among newly notified TB cases by age-group and bacteriological status.

J Lab Clin Med, 2002 Apr, 139(4), 244 - 50
Adherence of Helicobacter pylori to gastric epithelial cells and mucosal inflammation; Kaji T et al.; Adherence of Helicobacter pylori to the gastric epithelium is believed to be an important step in the induction of active inflammation of the mucosal layer . However, structural evidence showing a quantitative relationship between the adherence of H . pylori and severity of gastric mucosal inflammation is lacking . We therefore investigated the correlations between severity of gastritis and adherence of morphologically different forms of H . pylori . Fifty-seven biopsy specimens from the gastric bodies of patients with H . pylori-induced gastritis were examined . The severity of gastritis and the adherence and structure of H . pylori were determined with the use of light and scanning electron microscopy . We also investigated the ability of H . pylori organisms with different structural features to induce interleukin-8 secretion by human gastric adenocarcinoma (AGS) cells in vitro because production of interleukin-8 is related to H . pylori-associated gastritis . Furthermore, serum pepsinogen concentrations and cytotoxin-associated protein status in relation to adherence of H . pylori to the epithelial surface were examined . The results indicated that H . pylori organisms, which adhered firmly to the epithelial surface, were consistently long, tightly coiled bacilli . Histologically, those gastric mucosa samples with H . pylori firmly attached showed severe gastritis . H . pylori bacilli of greater length induced higher levels of interleukin-8 secretion . The serum pepsinogen I/II ratio showed a significant negative correlation with the grade of H . pylori adhesion (r = -0.401, P <.01) . We also noted a significant correlation between cytotoxin-associated protein status and the adherence of H . pylori (r = 0.344, P <.05) . A quantitative correlation was found between adherence of H . pylori and gastric inflammation . Both adherence and the induction of inflammation were found to be related to the structure of H . pylori.

BMC Gastroenterol . 2002 May 13;2(1):10.
Colonic tuberculosis mimicking Crohn's disease: case report; Chatzicostas C et al.; BACKGROUND: Intestinal tuberculosis is a rare disease in western countries, affecting mainly immigrants and immunocompromised patients . Intestinal tuberculosis is a diagnostic challenge, especially when active pulmonary infection is absent . It may mimic many other abdominal diseases . CASE PRESENTATION: Here, we report a case of isolated colonic tuberculosis where the initial diagnostic workup was suggestive of Crohn's disease . Computed tomography findings however, raised the possibility of colonic tuberculosis and the detection of acid-fast bacilli in biopsy specimens confirmed the diagnosis . CONCLUSIONS: In conclusion, this case highlights the need for awareness of intestinal tuberculosis in the differential diagnosis of chronic intestinal disease

Indian J Exp Biol, 2001 Dec, 39(12), 1227 - 34
Evaluation of immune responses directed against 30kDa secretory protein of Mycobacterium tuberculosis H37Ra complexed in different adjuvants; Yadav D et al.; Ability of different adjuvants to promote cell mediated immune responses towards 30 kDa secretory protein of Mycobacterium tuberculosis H37Ra was monitored by assessing the lymphocyte proliferation and IgG1/IgG2a subclass profile in mouse model . Six formulations, viz . poly lactide-co-glycolide (PLG) microspheres, dimethyldioctadecyl ammoniumbromide (DDA), liposomes, liposomes containing monophosphoryl lipid A and coated with alum (L-LIPA-AL) or without alum (L-LIPA) were evaluated in comparison to standard Freund's incomplete adjuvant (FIA) . Two adjuvant formulations of 30kDa-L-LIPA-AL and 30kDa-PLG showed maximum reactivity on VIIIth week post immunization (p.im) in terms of lymphoproliferation w.r.t . other adjuvant formulations . Both the vaccine formulations also exhibited a Th1 shift in terms of higher IgG2a response over IgGI . Flowcytometric analysis in the mesenteric lymph nodes (MLNs) of immunized animals revealed the capacity of 30kDa-PLG and 30kDa-L-LIPA-AL to activate T cell subsets like CD4 and CD8 T cells . The upregulation of B7 costimulatory molecules (B7-1 & B7-2) after immunization further proved the ability of the two vaccine formulations to activate antigen presenting cells . The immunostimulatory nature of the two formulations was also reflected in their capacity to reduce the bacilli load from the lungs of the experimentally infected mice . This study demonstrates PLG and L-LIPA-AL as potent adjuvants and their bioacceptibility and nontoxic nature make them suitable candidates for future subunit vaccine development against tuberculosis.

Curr Infect Dis Rep, 2002 Jun, 4(3), 249 - 256
Aerobic Gram-negative Bacillary Pneumonia; Parodi S et al.; Gram-negative bacilli (GNB) are a common cause of severe hospital-acquired pneumonia . Due to changes in the health care environment and selective antimicrobial pressure, these bacteria also are becoming a more common cause of pneumonia in venues outside of the traditional hospital setting and are increasingly resistant to antimicrobial agents . Risk factors for acquisition of GNB allow the clinician to efficiently identify patients who are likely to have pneumonia due to these pathogens . Available diagnostic techniques have a limited capacity to accurately detect GNB pulmonary infection . Yet, a pathogen specific diagnosis and knowledge of local resistance patterns are quintessential elements in formulating an effective treatment plan . This article reviews the epidemiologic characteristics, pathogenesis, and current management issues of GNB pneumonia.

New Microbiol, 2002 Apr, 25(2), 131 - 7
Diagnosis of paratuberculosis in naturally infected goats; Cerri D et al.; A survey was carried out to verify if an immunohistochemical method associated with agar gel immunodiffusion (AGID) will establish a firm diagnosis of caprine paratuberculosis . One hundred and thirty-six goats were tested by AGID for antibodies against Mycobacterium avium subsp . paratuberculosis at two different times: the first time 22 (19.1%) were positive and the second time 25 (18%) . One seronegative goat with severe diarrhea and 5 seropositive goats, two of which showing similar clinical signs, were sacrificed and necropsied . Samples were taken from small intestine, liver, spleen, mesenteric lymph nodes for bacteriological, histological and immunohistochemical examinations . M.a . paratuberculosis was isolated from intestine samples of 4 seropositive goats and from mesenteric lymph nodes of one seropositive goat; the microorganism was not isolated from samples of one seropositive and the seronegative animals . Ziehl Neelsen staining showed acid-fast bacilli in macrophages of the 5 seropositive animals and the immunohistochemical method for M . a . paratuberculosis detected bacterial antigen in the same samples.

Quintessence Int, 2002 May, 33(5), 376 - 82
Risk and prevention of transmission of infectious diseases in dentistry; Araujo MW et al.; Health care providers are at risk for infection with bloodborne pathogens, including hepatitis B virus, human immunodeficiency virus, and hepatitis C virus . Recommended infection control practices are applicable to all settings in which dental treatment is provided . Dentists remain at low risk for occupationally acquired human immunodeficiency virus . Dental health care workers, through occupational exposure, may have a 10 times greater risk of becoming a chronic hepatitis B carrier than the average citizen . Tuberculosis is caused by Mycobacterium tuberculosis . In general, persons suspected of having pulmonary or laryngeal tuberculosis should be considered infectious if they are coughing, are undergoing cough-inducing or aerosol-generating procedures, or have sputum smears positive for acid-fast bacilli . Although the possibility of transmission of bloodborne infections from dental health care workers to patients is considered to be small, precise risks have not been quantified by carefully designed epidemiologic studies . Emphasis should be placed on consistent adherence to recommended infection control strategies, including the use of protective barriers and appropriate methods of sterilization or disinfection . Each dental facility should develop a written protocol for instrument reprocessing, operatory cleanup, and management of injuries . Such efforts may lead to the development of safer and more effective medical devices, work practices, and personal protective equipment.

Dermatology, 2002, 204 Suppl 1, 15 - 20
Bactericidal activities of commonly used antiseptics against multidrug-resistant mycobacterium tuberculosis; Rikimaru T et al.; Seventeen clinical isolates of Mycobacterium tuberculosis were selected in order to study the bactericidal activities against drug-resistant M . tuberculosis . The effects of different antiseptics against multidrug-resistant M . tuberculosis (MDR-TB) were examined . Each of the test strains was cultured on the surface of an agar slant containing Lowenstein-Jensen medium . 0.05 ml of the bacillary suspension was poured into a test tube, and 0.45 ml of various antiseptics was added . After the bacilli had been exposed to the antiseptic solution with 2% human serum for various periods of incubation time, the antiseptic was inactivated by addition of 0.45 ml neutralizer, a mixture containing 10% Tween 80, 3% soybean lecithin and 0.5% sodium thiosulfate . As the results, povidone-iodine (PVP-I) at a concentration of 0.2% killed 99.9% or more of all strains tested within 30 s . All of the strains tested with PVP-I were killed almost completely within 60 s . There was no difference in bactericidal activities of PVP-I between standard strain H37Rv and MDR-TB . 99.9% or more of all strains tested were killed after exposure to 1.0% cresol for 60 s . In the case of cresol however, the exposure time of 30 s was not enough to get satisfactory effects . 2.0% glutaraldehyde needed 5 min to kill 99.99% or more of the bacilli tested, and 0.2% alkyldiaminoethylglycine hydrochloride required 60 min to do so . The results of bactericidal activities of common antiseptics against MDR-TB were similar to those against H37Rv . We conclude that the commercially available PVP-I product is a useful antiseptic against MDR-TB similar to other M . tuberculosis .

Vet Pathol, 2002 Mar, 39(2), 234 - 9
Clinical, microscopic, and molecular aspects of canine leproid granuloma in the United States; Foley JE et al.; Leproid granulomas from seven dogs in the United States were evaluated . Gross characteristics included nodular and ulcerated dermal and subcutaneous lesions primarily on the caudal aspects of the pinnae and to a lesser extent on the muzzle, face, and forelimbs . In all except one dog, there was complete regression of the lesions within 6 months, either with no therapy or after surgical resection . Cytology or histopathology revealed pyogranulomatous inflammation with few to many acid-fast mycobacterial bacilli within macrophages . The organisms could not be cultivated in vitro . DNA sequencing of part of the 16S ribosomal RNA gene region revealed 99-100% homology among fragments from five of these dogs and fragments from dogs in the south Pacific . This syndrome occurs in dogs in North America and the prognosis is excellent, in contrast to the prognosis for rapid-growing or tuberculous mycobacteriosis.

Int J Antimicrob Agents, 2002 May, 19(5), 361 - 70
New findings in beta-lactam and metronidazole resistant Bacteroides fragilis group; Fang H et al.; Beta-lactam antibiotics and 5-nitroimidazoles have been extensively used against anaerobic bacteria . However, antibiotic resistance is increasingly common among anaerobic Gram-negative bacilli . The classical mechanisms of resistance to beta-lactams are, (1) production of beta-lactamases; (2) alteration of penicillin-binding proteins (PBPs); and (3) changes in outer membrane permeability to beta-lactams . The 5-nitroimidazole molecule is a prodrug whose activation depends upon reduction of the nitro group in the absence of oxygen . Decreased uptake and altered reduction are believed to be responsible for metronidazole resistance . Five nim genes (A, B, C, D and E) have been identified in Bacteroides fragilis group spp . that confer resistance to 5-nitroimidazole antibiotics . Knowledge of the status and the mechanisms of resistance is critical for both the selection of antimicrobial therapy and the design of new antimicrobial agents . The purpose of this article is to review the mechanisms for and the prevalence of beta-lactam and metronidazole resistance in strains belonging to the B . fragilis group.

Chest, 2002 May, 121(5), 1486 - 92
Applying sputum as a diagnostic tool in pneumonia: limited yield, minimal impact on treatment decisions; Ewig S et al.; STUDY OBJECTIVES: We evaluated the role of sputum examination in the management of patients with community-acquired pneumonia (CAP) in a primary-care hospital without microbiologic laboratory facilities . Design and interventions: A diagnostic strategy using regular collection of sputum samples, Gram staining in a local laboratory, and mailing of samples to a commercial laboratory for culture analysis . SETTING: A 200-bed primary-care hospital without subspeciality physicians . PATIENTS: One hundred sixteen consecutive patients with a diagnosis of CAP were prospectively evaluated during a 12-month period . RESULTS: Of 116 patients, 42 patients (36%) were capable of producing a sputum sample . Age > or = 75 years (odds ratio {OR}, 0.4; 95% confidence interval {CI}, 0.18 to 0.93) and prior ambulatory antimicrobial treatment (OR, 3.2; 95% CI, 1.2 to 8.4) were independent predictors of sputum production . A delay in collection and processing of sputum samples of > 24 h was present in 31% and 39%, respectively . A delay in collection yielded an increased number of Gram-negative enteric bacilli and nonfermenters (44% vs . 7%, p = 0.056) . A delay in processing was associated with an increased number of Candida spp isolates (33% vs . 9%, p = 0.16) . The overall diagnostic yield was low (10 of 116 patients, 9%) due to a limited number of valid samples (n = 23 of 42 patients, 55%) and a limited number of definitely or probably positive samples on Gram's stain and culture (n = 10 of 42 patients, 24%) . Prior ambulatory antimicrobial treatment was associated with a reduction in diagnostic yield (14% vs . 56%, p = 0.09) . The impact of diagnostic results on antimicrobial treatment decisions was minimal, with antimicrobial treatment directed to diagnostic results in only one patient . CONCLUSIONS: We conclude that in this setting representative of primary-care hospitals in Germany, sputum had a low diagnostic yield and did not contribute significantly to patient management.

Curr Microbiol, 2002 Jun, 44(6), 450 - 9
Isolation and screening of brittlestar-associated bacteria for antibacterial activity; Strahl ED et al.; Many microbes associated with marine organisms have antimicrobial activity . We report the isolation of bacteria associated with Amphipholis gracillima that have broad-spectrum antibacterial activity against a number of common bacterial strains . Fifty-eight isolates of bacilli obtained from A . gracillima arm homogenates, from excised wound tissue, or from swabs of arm stumps exhibited 20-100% inhibition of one or more of 16 test bacteria at 35% salinity . Forty-one of the isolates were capable of 20-100% inhibition of one or more of 19 subject bacteria at 10% salinity at 37 degrees C . Three isolates, BE37, BE52, and BE53, exhibited the greatest range of antibacterial activity at both 10% and 35% salinity . Our results suggest that some of the bacteria associated with A . gracillima may provide the animal with chemical defenses against adverse bacterial infection . The water-soluble inhibitory chemicals produced by the bacteria could potentially function as antimicrobial compounds against human pathogenic bacteria.

Pediatr Dermatol, 2002 Mar-Apr, 19(2), 122 - 6
Lichen scrofulosorum: a rare manifestation of a common disease; Thami GP et al.; Tuberculids develop as hypersensitive immunologic reactions in the skin to an occult internal focus of tuberculosis . These eruptive lesions are due to hematogenous dissemination of bacilli in a host with a high degree of immunity against Mycobacterium tuberculosis . Although rare, these specific lesions are important diagnostic markers of tuberculosis . Lichen scrofulosorum is one of the recognized tuberculids, usually seen in children and young adults . We report three children with lichen scrofulosorum; in two children it developed during treatment of tuberculosis . The appearance of lichen scrofulosorum after initiation of treatment due to a probable increase in cell-mediated immunity is emphasized.

Poult Sci, 2002 Apr, 81(4), 585 - 9
Oral lavage with polyethylene glycol reduces microbial colonization in the gastrointestinal tract of broilers; Farhat A et al.; Oral lavage of 7-wk-old broiler chickens with polyethylene glycol (PEG) was conducted to induce rapid evacuation of the gastro-intestinal tract (GIT) before processing . Three groups of birds were fed ad libitum (FF), subjected to feed withdrawal 12 h before processing (FW), or received 115 mL of PEG by oral lavage 3 h before processing (PEG(L)) . The crop, a section of small intestine, and the right cecum were aseptically collected for determination of contents and Gram-negative bacilli (GNB) counts . The PEG(L) effectively cleansed the upper GIT and intestine of treated birds within 3 h before processing . However, the ceca were not completely empty for all the birds subjected to PEG(L) . The PEG(L) group had a significantly lower crop and cecal GNB concentrations than the FW group . Birds on FW and PEG(L) treatments had significantly lower GNB concentrations in the small intestine than FF birds . Polyethylene glycol resulted in complete evacuation of the upper GIT, as well as the intestine, within 3 h of use, and significantly reduced GNB concentrations.

Am J Infect Control, 2002 May, 30(3), 158 - 64
Hospital-acquired meningitis in patients undergoing craniotomy: incidence, evolution, and risk factors; Reichert MC et al.; OBJECTIVES: To establish the incidence of postcraniotomy meningitis, identify etiologic agents, assess patients' medical progress in relation to both length of hospitalization and mortality, and analyze risk factors . MATERIAL AND METHODS: This study was developed at Hospital Sao Paulo, a tertiary university hospital, between August 1995 and January 1998 . We conducted a case-control trial, in which 50 pairs of patients were matched . RESULTS: An 8.9%-postcraniotomy meningitis incidence was found . Gram-negative bacilli were the most common etiologic agents isolated . Mortality among the patients was 30% . Mean hospital stay for the patients was 42.9 +/- 22.1 days; for the controls, mean hospital stay was 19.0 +/- 11.4 days (P =.00001) . Although several risk factors were identified by univariate analysis, including postoperative external ventricular shunt (OR = 2.92, CI 95% = 1.245-6.865, P =.014), remote site infection (OR = 2.85, CI 95% = 0.995-8.173, P =.051), and repeat operation (OR = 5.02, CI 95% = 1.569-16.066, P =.007), only repeat operation remained in the multivariate analysis model (OR = 3.68, CI 95% = 1.158-11.700, P =.027) . CONCLUSION: Postcraniotomy meningitis resulted in a high mortality rate and a longer hospital stay, with repeat operation identified among the risk factors.

J Basic Microbiol, 2002, 42(2), 105 - 10
Toxicity of Bacillus thuringiensis and B . sphaericus to laboratory populations of Drosophila melanogaster (Diptera: Drosophilidae); Khyami-Horani H; A total of 65 samples, consisting of 8 sample types, collected from the Jordan Valley, were examined for the presence of Bacillus thuringiensis and B . sphaericus and for their toxicity against the larvae of the fruit fly Drosophila melanogaster . The frequency of samples containing toxic aerobic spore-forming bacilli was 12%; of which 21.7% belonged to B . thuringiensis and 17.4% to B . sphaericus . The B . thuringiensis populations consisted of 5 serogroups: thuringiensis (H1), entomocidus (H6), pakistani (H13), autoagglunated, in addition to a new serotype . The B . sphaericus population consisted of 3 serogroups, and belonged to serovars H5, H9, and H13.All B . thuringiensis and B . sphaericus local isolates, in addition to the reference strains B . thuringiensis kuristaki, and B . thuringiensis israelensis, showed high toxicity towards 3(rd) instar larvae of D . melanogaster . The toxic concentrations ranged between 2.0 x 10(6) and 4.4 x 10(7) viable spores ml(-1).

Childs Nerv Syst, 2002 Apr, 18(3-4), 118 - 23 Epub 2002 Mar 22.
Tuberculous brain abscess: clinical presentation, pathophysiology and treatment (in children); Kumar R et al.; BACKGROUND: Tubercular brain abscess (TBA) is a rare manifestation of CNS tuberculosis . It is characterised by an encapsulated collection of pus, containing viable tubercular bacilli without evidence of tubercular granuloma.PRESENTATION AND HISTORY: Patients may present with features of raised intracranial pressure and focal neurological deficit commensurate with the site of the abscess . A history of pulmonary tuberculosis may be present, as documented in one of our six cases; three of our six children developed TBA despite 3-weeks to 12-month courses of antitubercular chemotherapy prescribed for post-TBM hydrocephalus.DIAGNOSIS: Contrast CT head, MRI, MR spectroscopy is helpful in making the diagnosis and planning the treatment . TBA may be unilocular or multilocular on contrast CT scan . A relatively long clinical history and an enhancing capsule with thick wall are suggestive of TBA . Pyogenic abscess, however, has a thin rim on contrast CT . The capsule of TBA is formed of vascular granulation tissue containing acute and chronic inflammatory cells, particularly polymorphs . Proof of tubercular origin must be demonstrated either by presence of acid fast bacilli in culture or staining of pus or wall.TREATMENT: Treatment options include simple puncture, continuous drainage, fractional drainage, repeated aspiration through a burr hole, stereotactic aspiration and total excision of the abscess . Total excision usually becomes necessary in multilocular noncommunicating and thick-walled abscesses . Antitubercular therapy is the mainstay of management . The development of fulminant tubercular meningitis is sometimes problematic following surgical excision of TBA, as seen in one of our four operated cases . Mortality is reported to be high despite progress in treatment, while five of the six children treated by us responded well to the treatment.

Ann Plast Surg, 2002 May, 48(5), 557 - 61
Nonhealing ulcerative mass of the elbow: do not forget tuberculosis; Ayhan S et al.; Skeletal tuberculosis is a rare condition and is seen in only 1 to 2% of all cases of tuberculosis . The authors present a 69-year-old white woman with tuberculosis of the left elbow joint whose disease was suspicious for synovial sarcoma . Skeletal tuberculosis usually conflicts with neoplastic or inflammatory diseases . One should remember the possibility of bone-joint tuberculosis and send biopsy specimens for culture to determine the presence of acid-fast bacilli . However, neither culture nor polymerase chain reaction nor histological examination may be capable of showing the bacilli . Only a suspicion may be enough . The treatment protocol is usually a combination of surgical debridement and multidrug antituberculous chemotherapy.

Curr Opin Infect Dis, 2001 Dec, 14(6), 711 - 5
Evidence for antibiotic cycling in control of resistance; Pujol M et al.; There is growing concern about the increasing rates of resistance among human pathogens . These high rates of resistance, which are a consequence of antibiotic misuse, highlight the need for a thorough revision of strategies that deal with the proper use of antibiotics . The so-called 'optimal use of all antibiotics', restriction guidelines and use of a combination of antibiotics are well established strategies in most hospitals but new potential strategies, such as dosage optimization and antibiotic cycling, require further study and evaluation . Cycling antibiotic therapy consists of the scheduled replacement of one antibiotic for another, in order to avoid the development of bacterial resistance . Initial studies suggest that this strategy could be useful in reducing the rates of bacterial resistance as well as the incidence of nosocomial infections caused by Gram-negative bacilli in intensive care unit patients . This strategy, however, does not prevent antibiotic misuse and needs to be applied in a complete antibiotic policy program.

Clin Orthop, 2002 May, (398), 107 - 13
Tuberculosis of the foot and ankle; Dhillon MS et al.; Tuberculosis of the foot and ankle is an uncommon presentation of skeletal tuberculosis . The uncommon site, lack of awareness, and ability to mimic other disorders clinically and on radiographs, leads to diagnostic and therapeutic delays . In the early stages and when the disease is limited to bone, medical treatment leads to excellent healing and limited residual disabilities . Joint involvement occurs because of spread from a periarticular bony focus, and in the midfoot, the disease may spread to involve all the interconnected joints, leading to a stiff foot and residual deformities . Of 74 cases reviewed, the calcaneus was involved most commonly, followed by infection of the midtarsal, Lisfranc joints, and ankle . The most common radiologic finding is that of osteoporosis, which may be intense: cancellous bone involvement may present as a cystic lesion with or without sequestrum . Computed tomography scans and magnetic resonance imaging are helpful examinations . Because the disease is paucibacillary, a positive acid fast bacilli culture is rare and the diagnosis usually is confirmed by obtaining granulomatous tissue on biopsy . The treatment basically is medical, with surgical intervention being reserved for patients with intractable disease or as a salvage procedure for patients with deformed hindfoot joints.

Appl Microbiol Biotechnol, 2002 Mar, 58(4), 547 - 53
Bacillus naphthovorans sp . nov . from oil-contaminated tropical marine sediments and its role in naphthalene biodegradation; Zhuang WQ et al.; A Bacillus sp., designated as strain MN-003, was isolated as the dominant cultivatable naphthalene-degrading organism from oil-contaminated tropical marine sediments . Strain MN-003 is strictly aerobic, rod-shaped, Gram-positive, catalase positive, oxidase negative, and forms endospores . Strain MN-003 grew at salinities ranging from 0.28 to 7.00% and temperatures ranging from 15 to 41 degrees C . Phylogenetic analyses reveal that strain MN-003 is most similar to Bacillus sp . VAN14, with a 16S rRNA sequence identity of 97.9% . Based on taxonomic and 16S rRNA data, strain MN-003 was named Bacillus naphthovorans sp . nov . When grown with naphthalene as sole carbon source, strain MN-003 had a maximal specific growth rate (mu(max)) of 0.32 +/- 0.03 h(-1), and a half-saturation constant (K(S)) of 22.3 +/-4.2 microM . A batch study of the tropical marine sediments enriched with naphthalene showed that cells of the Bacillus genus grew to become dominant members of the microbial community . The bacilli comprised 39.5 +/- 6.5% of the microbial fraction after 20 days of enrichment.

Infect Immun, 2002 May, 70(5), 2566 - 75
Glucan is a component of the Mycobacterium tuberculosis surface that is expressed in vitro and in vivo; Schwebach JR et al.; The outermost layer of Mycobacterium tuberculosis is composed primarily of two polysaccharides, glucan (GC) and arabinomannan . To analyze the surface polysaccharide composition of M . tuberculosis, we generated a monoclonal antibody (MAb) that binds M . tuberculosis GC and is known as MAb 24c5 . Immunofluorescence and whole-mount immunoelectron microscopy indicated that GC is on the outermost portion of the bacteria . M . tuberculosis strains Erdman and CDC 1551 were analyzed for their ability to bind MAb 24c5 after in vitro growth in media with and without the detergent Tween 80 . MAb 24c5 bound to Erdman and CDC 1551 at all culture times with only slightly greater apparent affinity after extended culture in the absence of Tween 80, indicating that a stable amount of GC polysaccharide antigen is associated with the cell surface of M . tuberculosis . An enzyme-linked immunosorbent assay indicated that GC is antigenically similar to glycogen, and the amount of GC antigen increased in the media of M . tuberculosis cultures grown either with or without the detergent Tween 80 . Other nontuberculosis mycobacteria have antigenically similar GCs on their surfaces after in vitro growth . Inoculation of mice with live bacilli but not inoculation with dead bacilli elicited a strong antibody response to GC consistent with production of this antigen in vivo . Our results provide a more comprehensive picture of the M . tuberculosis cell envelope and the conditions that allow expression of M . tuberculosis GC.

Zhonghua Jie He He Hu Xi Za Zhi, 2002 Jan, 25(1), 21 - 4
{Experimental study on prevention of diabetes complicated with tuberculosis by Mycobacterium vaccae}; Chen H et al.; OBJECTIVE: To study if Mycobacterium vaccae has any effect in prevention of rats with diabetes from tuberculosis . METHODS: Wistar rats with diabetes which were produced by injection of streptozotocin one week before were divided into four groups . Mycobacterium vaccae was injected into Wistar rats with diabetes and one month later H(37)R(v) was injected intravenously . The rats were sacrificed six weeks later to observe pathomorphologic changes in lung tissues . Indexes for evaluation included quantitative culture of Mycobacterium tuberculosis of lung tissue, tubercular lesion, count of Mycobacterium tuberculosis in alveolar macrophages and histopathological changes . RESULTS: Pathological change index of tubercular lesion, quantitative culture of Mycobacterium tuberculosis in lung tissue, count of Mycobacterium tuberculosis in alveolar macrophages were 2.5 +/- 0.7, (4.1 +/- 0.6) x 10(4) cfu, 2.6 +/- 0.9 respectively in group injected with Mycobacterium vaccae, while 3.3 +/- 0.5, (9.9 +/- 1.0) x 10(4) cfu, 7.2 +/- 0.7 respectively in the control group . Statistically significant differences were found between the two groups (P < 0.05, < 0.01, < 0.01 respectively) . The histopathological changes of lung tissue in group of diabetes complicated with tuberculosis revealed that the lesions were predominant with necrosis, while in group injected with Mycobacterium vaccae, the lesions showed proliferation mainly . This indicated that Mycobacterium vaccae could decrease the damage to the lung tissue of the rat with diabetes by tubercle bacilli . CONCLUSION: Mycobacterium vaccae might play a role in prevention of rat with diabetes from tuberculosis.

Mikrobiol Z, 2002 Jan-Feb, 64(1), 31 - 6
{Screening of collagenase and keratinase producers}; Ivanko OV et al.; The study of the capacity of 310 strains of microorganisms from different taxonomic groups (40 bacilli, 43 yeast, 105 streptomycetes, 12 micromycetes) to hydrolyze collagen and keratin allowed to establish that the highest level of collagenase (KA) and keratinase (KerA) activity is inherent in representatives of streptomycetes . Two strains of Streptomyces sp.--1349 and 1382 with the highest KA and KerA indices--1.9 and 1.85 un./mg of protein, respectively, have been chosen . It has been established that collagenase activity in the medium without adding the inducers decreases 4.76 times, while that of keratinase--5.71 times, i.e . the above enzymes are inducible . The investigation of the spectrum of activities has demonstrated that the both strains possess low level of the general proteolytic and elastase activities and high level of collagenase and keratinase activities . Partial purification of the enzyme complex of Streptomyces sp . 1349 by the successive precipitation by ammonium sulphate with 30, 60 and 80% saturation and a single precipitation by ammonium sulphate with 80% saturation helped to increase the level of KA 5.6-5.9 times, and that of KerA--4.2-4.5 times.

Hunan Yi Ke Da Xue Xue Bao, 1999, 24(2), 150 - 2
{Intestinal bacterial translocation in patients with biliary tract diseases}; Yu M et al.; To determine whether intestinal bacterial translocation occurs in man, the authors cultured the tissues of mesenteric lymph nodes(MLNs), bile, portal and peripheral venous blood obtained prior to and during operation in 35 patients with biliary diseases . No positive peritoneal swab, portal blood and peripheral blood culture were shown . Thirteen of 20 patients with biliary obstruction (65%) had bacteria in their MLNs, while no positive MLN culture was found in 12 patients without biliary obstruction . The most common bacteria recovered from the MLNs were Gram-negative enteric bacilli . Thus, it is considered that extrahepatic biliary obstruction may induce intestinal bacterial translocation in man.

Int J Tuberc Lung Dis, 2002 Mar, 6(3), 270 - 2
Washing of new microscopic glass slides in dichromate solution does not influence sputum AFB smear results; Selvakumar N et al.; SETTING: Microscopy centres in Tiruvallur District, Tamil Nadu, India, implementing DOTS . OBJECTIVE: To know whether washing new glass slides in dichromate solution is essential for effective sputum acid-fast bacilli microscopy . METHODS: Two direct smears were prepared from each of 1750 sputum samples . One was made on dichromate solution-cleaned new glass slides and the other was made on unwashed new glass slides . The smears were blinded and examined . RESULTS: Of the 1750 specimens, 13.5% and 13.08% were positive for AFB using washed and unwashed slides, respectively (P = 0.12) . The concordance between these two (including one grade above and one below) was 98.7% . CONCLUSION: Washing of new glass slides in dichromate solution is not essential for AFB microscopy.

Int J Tuberc Lung Dis, 2002 Mar, 6(3), 222 - 30
Optimal tuberculosis case detection by direct sputum smear microscopy: how much better is more?
Van Deun A, Salim AH, Cooreman E, Hossain MA, Rema A, Chambugonj N, Hye MA, Kawria A, Declercq E.
SETTING: A tuberculosis control project in Bangladesh . OBJECTIVE: To define the efficiency of numbers of microscopic fields screened and the sputum collection scheme used for diagnostic smear examination . DESIGN: Quality controllers noted cumulative numbers of acid-fast bacilli per 100 fields screened . The incremental diagnostic yield of different sputum sampling strategies was determined . Doubtful series were re-checked and/or further samples examined . RESULTS: Acid-fast bacilli were found in 99.6% of 1412 positive and in 79.3% of 576 scanty slides in the first 100 fields . Examination of a third specimen yielded a maximum of 2.7% positives incrementally . The most efficient strategy, using three morning specimens, yielded 94.2% positives on the first and 1.0% on the third sputum; although 10% of suspects did not return, only 1.5% of the positives were among them and more cases were confirmed and treated . The positive predictive value of a single positive or scanty smear was very high (99.2%) . CONCLUSIONS: Reading more than 100 fields per smear or examining a third sputum has insufficient marginal returns to justify the workload . Examining morning samples only is more efficient, and their collection does not necessarily inconvenience patients . Treatment can be started on the basis of one positive smear . Provided that a well functioning system of smear-microscopy quality control is in place, we propose a strategy based on examination of two morning sputum samples for negative suspects, with the diagnosis based on a single positive result.

Int J Tuberc Lung Dis, 2002 Feb, 6(2), 91 - 7
Relation of grading of sputum smears with clinical features of tuberculosis patients in routine practice in Sudan; El-Sony A et al.; SETTING: Health services in Khartoum, Red Sea, Gadarif, Kassala, Bahr el Jebel, Gezira and North Kordofan States of Sudan . OBJECTIVE: To investigate the association of presenting symptoms and clinical history with grade of sputum smear positivity in tuberculosis patients . METHODS: The 5338 tuberculosis suspects aged 15 to 49 years who consulted between March 1998 and March 1999 underwent sputum smear microscopy, with smears graded by a standardised method . Patients who gave consent had a structured interview concerning the nature and duration of their symptoms and whether they had a sick relative at home . Statistical analysis determined the association of symptoms and home characteristics with the results of smear examination . RESULTS: A total of 514 suspects (9.6%) demonstrated acid-fast bacilli: 45 (8.8%) grade +1, 167 (32.5%) grade +2 and 302 (58.8%) grade +3 . Most of the patients had a duration of symptoms of 6-9 weeks, while 100 had symptoms of more than one year's duration, many (n = 47) with other chronic respiratory diseases . Multiple chest symptoms were positively, and duration of symptoms negatively associated with grade of positivity (P = 0.018 and 0.028, respectively) . A high grade of smear positivity (+3) and the longest duration of symptoms, among those reporting symptoms of less than one year's duration, were significant predictors of the presence of a sick household member (OR = 1.99, 95%CI 1.12-3.55; OR = 1.004, 95%CI 1.000-1.008) . CONCLUSION: This study demonstrates a relation between the grade of smear positivity and the clinical features of tuberculosis patients.

Int J Tuberc Lung Dis, 2002 Feb, 6(2), 150 - 4
Mycobacterium tuberculosis typing: usefulness of DRE-PCR to confirm cross-contamination in the mycobacteriology laboratory of a general reference hospital for AIDS; Filho LA et al.; In this study two molecular typing methods, a simple double repetitive element PCR-based assay and the standardized restriction fragment length polymorphism (RFLP), were used to confirm cross-contamination in the mycobacteriology laboratory . Clinical specimens from 12 patients, submitted for acid-fast bacilli stain smear and processed for culture in Lowenstein-Jensen on the same day, resulted in positive bacterioscopy (+++) and confluent growth only for one of the patients . The specimens from all the other patients but two were smear-negative and culture-positive, with one or two colonies . None of them had clinical symptoms and radiological findings for active tuberculosis (TB) . The suspicion of false-positive cultures arose when a health care worker who had had a PPD skin test conversion, claimed to be healthy and had no TB symptoms, was found to have a positive sputum culture . DRE-PCR demonstrated that all nine cultures typed belonged to one cluster, further confirmed by RFLP . Although DRE-PCR has been found to be poorly reproducible, it has enough discriminatory power to be useful for rapid epidemiological investigation in selected settings.

Int J Tuberc Lung Dis, 2002 Feb, 6(2), 143 - 9
A retrospective study for the outcome of pulmonary resection in 49 patients with multidrug-resistant tuberculosis; Park SK et al.; BACKGROUND: Outcome of treatment using only chemotherapy for multidrug-resistant tuberculosis is usually considered not to be satisfactory . The combination of chemotherapy with surgical treatment has been producing higher acid-fast bacilli (AFB) negative sputum conversion and longer survival rates . This treatment strategy may be the solution for patients with multidrug-resistant tuberculosis . MATERIALS AND METHODS: A retrospective review was performed of the medical records and laboratory findings of 49 patients with multidrug-resistant tuberculosis among 130 patients who underwent pulmonary resection for pulmonary tuberculosis between January 1995 and December 1999 at National Masan Tuberculosis Hospital, Korea . RESULTS: The mean number of drugs to which the patients were resistant was 4.5 . Patients had a mean age of 35 years . Cavitary lesions on plain chest X-ray were shown in 43/49 patients (87.8%); 31 had positive sputum cultures preoperatively (63.3%) . The surgical techniques used were as follows: 12 pneumonectomies, 28 lobectomies, seven lobectomies with segmentectomies or wedge resections, one wedge resection and one cavernoplasty . The AFB negative sputum conversion rate was 93.5% with continuous postoperative chemotherapy . There were no deaths after surgery . Postoperative complications that developed were six cases of air leakage over a week, one of postoperative bleeding and one of wound infection . CONCLUSIONS: This study demonstrated the effectiveness of pulmonary resection with postoperative chemotherapy in cases of multidrug-resistant tuberculosis . Although there are different ideas about indications for surgery and the length and content of postoperative drug regimens, pulmonary resection should be considered an effective measure in combination with chemotherapy for treatment of multidrug-resistant pulmonary tuberculosis.

Int J Tuberc Lung Dis, 2002 Jan, 6(1), 47 - 54
Quality assessment of sputum transportation, smear preparation and AFB microscopy in a rural district in Malawi; Mundy CJ et al.; SETTING: Ntcheu District, Central Region of Malawi . OBJECTIVES: To assess 1) the feasibility of introducing simple internal quality control procedures for acid-fast bacilli (AFB) microscopy, and 2) the quality of the district sputum smear microscopy service . DESIGN: A simple internal quality control system was piloted in which district laboratory staff assessed: 1) specimen suitability, 2) time between sputum submission and smear examination, 3) smear preparation and staining, and 4) microscopy . Actual times for processing specimens were compared with recommended times . External quality validation was carried out . RESULTS: Of 4805 sputum specimens: 1) documentation was complete in 95%, 2) 93% reached the laboratory within 7 days of collection, 3) 96% of smears were well prepared and stained, and 4) 97% concordance (96.4% smear-positive and 97.6% smear-negative) was demonstrated when 208 smears were re-examined by a second technician . The aggregate index of reliability was 86% . The mean time spent on microscopic examination was 3.8 minutes, compared with the recommended time of 10 minutes . When all smears from 164 patients were assessed externally, 98.2% concordance (98.1% smear-positive and 98.2% smear-negative) was demonstrated . False smear-negative and smear-positive rates were less than 2% each . CONCLUSION: District laboratory staff were able to incorporate simple quality control procedures for AFB microscopy into their routine practice, resulting in a reliable service . The lessons learnt are widely relevant and potentially useful for implementation of a national quality assurance scheme.

Mol Microbiol, 2002 Feb, 43(3), 717 - 31
Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling; Betts JC et al.; The search for new TB drugs that rapidly and effectively sterilize the tissues and are thus able to shorten the duration of chemotherapy from the current 6 months has been hampered by a lack of understanding of the metabolism of the bacterium when in a 'persistent' or latent form . Little is known about the condition in which the bacilli survive, although laboratory models have shown that Mycobacterium tuberculosis can exist in a non-growing, drug-resistant state that may mimic persistence in vivo . Using nutrient starvation, we have established a model in which M . tuberculosis arrests growth, decreases its respiration rate and is resistant to isoniazid, rifampicin and metronidazole . We have used microarray and proteome analysis to investigate the response of M . tuberculosis to nutrient starvation . Proteome analysis of 6-week-starved cultures revealed the induction of several proteins . Microarray analysis enabled us to monitor gene expression during adaptation to nutrient starvation and confirmed the changes seen at the protein level . This has provided evidence for slowdown of the transcription apparatus, energy metabolism, lipid biosynthesis and cell division in addition to induction of the stringent response and several other genes that may play a role in maintaining long-term survival within the host . Thus, we have generated a model with which we can search for agents active against persistent M . tuberculosis and revealed a number of potential targets expressed under these conditions.

Pneumonol Alergol Pol, 2001, 69(9-10), 564 - 7
{Tuberculous peritonitis as still probable cause of ascites}; Ucinska R et al.; A 38-year-old man was admitted to hospital because of ascites, fever, weakness, abdominal pain . Chest x-ray revealed old post-tuberculous opacities in the left subclavicular region and fluid in left pleural cave . Cirrhosis of the liver and malignant disease were suspected . Treatment with antibiotics and evacuation of pleural fluid caused short-lasting improvement . After 6 weeks of diagnostic procedures parenchymal infiltrations in left lung were confirmed . In sputum tuberculous bacilli were found . Tuberculous peritonitis was confirmed histopathologically . Antituberculous treatment was successful.

Nihon Kokyuki Gakkai Zasshi, 2002 Jan, 40(1), 35 - 9
{A case of mixed pulmonary infection with Mycobacterium tuberculosis and M . intracellulare due to familial prevalence}; Kobashi Y et al.; A 76-year-old woman was admitted to our hospital because of an abnormal shadow on chest radiography and a suspected familial prevalence of pulmonary tuberculosis . Her son had previously been admitted to our hospital with pulmonary tuberculosis and tuberculous pleuritis . In the present case, a diagnosis of pulmonary tuberculosis and atypical pulmonary mycobacteriosis was made on the basis of detection of both Mycobacterium tuberculosis and M . intracellulare several times in a sputum culture of anti-fast bacilli . Restriction fragment length polymorphism (RFLP) analysis was then performed for both the woman and her son . The RFLP analysis revealed the same pattern in both mother and son, and confirmed that this was a case of mixed pulmonary infection passed from son to mother.

Comp Med, 2001 Apr, 51(2), 171 - 5
Atypical piscine mycobacteriosis in Japanese medaka (Oryzias latipes); Sanders GE et al.; Japanese medaka, (Oryzias latipes), small, freshwater, tropical cyprinodonts, are principally used for toxicologic and carcinogenicity assays, but are finding more applications in developmental genetic and biological research . An increase in mortality began in brood stock of adult medaka that had been shipped and housed separately by sex . Initially, mortality averaged one fish daily and began in females two weeks after they were received . Cohabitation began eight weeks after arrival . After four to six weeks of cohabitation in different spawning aquaria, mortality was observed in males . Clinical signs of disease included loss of scale luster and color, with subsequent blanching of dorsal flank musculature, small raised nodules on various external surfaces, emaciation, fraying of fin tips, and equilibrium disturbances . Histologic examination of affected adults revealed multi-organ granulomatous inflammation with intracellular acid-fast bacilli . Specimens from 46 juvenile medaka that were spawned from affected adults, were submitted for culture and histologic evaluation . Of 18 fish, two had lesions similar to those of adults . The organism isolated from the remaining fish was identified as Mycobacterium fortuitum . Due to atypical rapid progression of disease, spread of M . fortuitum to progeny, and poor prognosis, the entire colony was euthanized.

Niger Postgrad Med J, 2001 Dec, 8(4), 199 - 202
Tuberculosis myocarditis: a case report; Danbauchi SS et al.; A 35-year old man presented with fever, weight loss, drenching night sweats and symptoms of cardiac failure for three months . Examination revealed wasting, peripheral oedema, bilateral pleural effusion and constrictive pericarditis . A diagnosis of constrictive pericarditis with bilateral pleural effusion probably due to tuberculosis was made . Human immunodeficiency virus antibodies and six sputum for acidfast bacilli were negative . Electrocardiograph revealed low voltages globally and echocardiography showed global myocardial hypokinesia . He had pericardiectomy, pericardial and pleural histology was non-specific inflammatory reaction but myocardial histology showed granulomatous changes of tuberculous myocarditis . We suggest that in experienced hands myocardial biopsy could be useful in making the diagnosis.

J Clin Microbiol, 2002 Apr, 40(4), 1219 - 24
Relatively alcohol-resistant mycobacteria are emerging pathogens in patients receiving acupuncture treatment; Woo PC et al.; Acupuncture has been gaining popularity as a form of alternative medicine . In the past, only blood-borne viruses and anecdotal reports of bacterial infections have been associated with acupuncture . We report on four patients with mycobacterial infections complicating acupuncture who were encountered in a 2-year period . All had clinical and/or radiological lesions at acupuncture point- and meridian-specific locations . There was no other history of trauma or other clinical foci of infections, and the chest radiographs were normal . Histological studies of biopsy specimens of all four patients showed changes compatible with chronic inflammation, with granulomatous inflammation present in three patients and acid-fast bacilli present in two . Conventional biochemical tests and whole-cell fatty acid analysis for identification were inconclusive for all four nonpigmented mycobacteria recovered from tissue biopsies . 16S rRNA gene sequencing showed that the strains from two patients were Mycobacterium chelonae and that those from the other two were Mycobacterium nonchromogenicum . Alcohol resistance assay using the quantitative suspension test revealed that all four strains showed prolonged survival in 75% alcohol compared to other skin flora . Mycobacterial infections transmitted by acupuncture are an emerging problem . A high index of suspicion is essential to recognize this clinical syndrome, and strict implementation of proper infection control guidelines for acupuncture is mandatory.

Infect Control Hosp Epidemiol, 2002 Mar, 23(3), 141 - 4
Preventing nosocomial transmission of pulmonary tuberculosis: when may isolation be discontinued for patients with suspected tuberculosis?
Siddiqui AH, Perl TM, Conlon M, Donegan N, Roghmann MC.
OBJECTIVE: The Centers for Disease Control and Prevention and the American Thoracic Society recommend obtaining cultures of at least three sputum specimens for acid-fast bacilli (AFB) from patients in whom tuberculosis (TB) is suspected . On the basis of this, most hospitals isolate patients with suspected TB for 3 days or more until three smear (not culture) results are negative . Our objective was to evaluate the predictive value and sensitivity of these smears . DESIGN: Observational study . SETTING: Four urban medical centers . METHODS: The posttest probability of TB given sequential negative AFB smears from 274 patients isolated for suspected TB and the sensitivity of sequential AFB smears from 209 patients with positive results on culture for pulmonary TB were measured . RESULTS: The posttest probabilities of having TB given one, two, and three negative AFB smears were low: 1.1% (3 of 265; 95% confidence interval {CI95}, 0.23% to 3.27%), 0.4% (1 of 262; CI95 0% to 2.1%), and 0% (0 of 260; CI95, 0% to 1.4%), respectively . Among the 209 patients with positive results on culture for pulmonary TB, 169 (81%) had an expectorated sputum specimen sent, of which 91 (54%) were positive for AFB . Forty (24%) of the 169 patients had a second expectorated sputum specimen sent after the results of the first specimen were negative; only 6 (15%) of these had positive AFB smears . None of the 10 patients in whom the first two expectorated sputum samples yielded an AFB smear without an organism had a third AFB smear that was positive . CONCLUSION: Unless there is high clinical suspicion of pulmonary TB in a specific patient, the use of three AFB smears on expectorated sputa is a rational approach to discontinuing isolation for patients with suspected TB.

Acta Cytol, 2002 Mar-Apr, 46(2), 386 - 8
Cytodiagnosis of erythema nodosum leprosum . A case report; Anshu et al.; BACKGROUND: The application of cytology in leprosy has been restricted to the evaluation of morphologic and bacterial indices by slit skin smears to facilitate diagnosis of cases according to the Ridley-Jopling scale . Isolated reports have now documented the use of fine needle aspiration cytology (FNAC) in the diagnosis of leprosy . CASE: A 45-year-old male presented with the abrupt onset of multiple nodular eruptions all over the body . The clinical diagnosis was Sweet's syndrome . FNAC showed numerous neutrophils in a background of foamy macrophages . Special stains revealed the presence of a large number of fragmented acid-fast bacilli in the smears . A diagnosis of erythema nodosum leprosum (ENL) was made on FNAC . CONCLUSION: The presence of neutrophils in a characteristic milieu of foamy macrophages is seen in lesions of ENL . Such a picture should prompt the cytologist to use a modified Ziehl-Neelsen stain to demonstrate acid-fast bacilli, as ENL can present as an acute episode in patients without a previous diagnosis of leprosy.

Acta Crystallogr D Biol Crystallogr, 2002 Apr, 58(Pt 4), 615 - 21 Epub 2002 Mar 22.
Specificity of TRAP-RNA interactions: crystal structures of two complexes with different RNA sequences; Hopcroft NH et al.; The trp RNA-binding attenuation protein (TRAP) regulates expression of the tryptophan biosynthetic genes in bacilli by binding to the leader region of the nascent trp operon mRNA . When activated by binding tryptophan, the 11-subunit circular TRAP molecule binds to a target sequence consisting of 11 (G/U)AG repeats, separated by two or three variable 'spacer' nucleotides . Reported here are two crystal structures of TRAP bound to RNAs containing 11 GAG repeats separated by UU and CC spacer nucleotides, determined at 1.75 and 2.50 A resolution, respectively . These show the spacer regions of the RNA molecules to be highly flexible, making no direct hydrogen-bonding contacts with the protein . Comparison of these structures with the previous structure of TRAP bound to (GAGAU)(10)GAG RNA, in which the spacer nucleotides stack with each other close to the protein surface, shows that the RNA can adopt different conformations depending on the sequence of the spacer regions . This gives insight into the structural basis of the specificity of TRAP and into the mechanism of binding.

Tuberculosis (Edinb), 2002, 82(1), 31 - 6
Interaction of Mycobacterium tuberculosis with human respiratory epithelial cells (HEp-2); Reddy VM et al.; SETTING: Mycobacterium tuberculosis (MTB) is known to adhere to, invade, multiply inside, and modulate respiratory epithelial cell functions . However, the mode and the molecules involved in the interaction of MTB with respiratory cells are not known . OBJECTIVE: To study post-adherence morphological changes in human respiratory epithelial cells and identify MTB components involved in the binding to these cells . DESIGN: The mechanism of interaction of MTB human respiratory epithelial cells (HEp-2) was investigated by incubating MTB with HEp-2 cells, whereupon the morphological changes were examined under scanning electron microscopy . MTB components involved in the interaction with HEp-2 cells were identified by probing Western blots of mycobacterial sonicates with biotinylated HEp-2 extracts . RESULTS: MTB readily bound to the surface of HEp-2 cells . Adherence of tubercle bacilli on the surface caused membrane perturbation resulting in increased surface membrane projections, particularly towards the margins of the cells . Long thin epithelial membrane projections formed loops around the bacilli holding them on the surface . Membrane projections fused to form a web in which the bacilli were entrapped . Concomitant membrane cavitations seen beneath the site of attachment of the bacilli conceivably caused internalization of the organisms . At least five MTB proteins ( approximately 112, 35, 28, 21 and 15 kDa) specific for HEp-2 cells were identified . Of these, the 28 kDa protein was the predominant HEp-2-binding protein and was found to be heparin-binding hemagglutinin (HBHA) . While M . bovis BCG strain showed the same five HEp-2-binding proteins, M . smegmatis, displayed only one (31 kDa) HEp-2-specific protein . CONCLUSION: MTB binds to HEp-2 cells through multiple proteins, and the binding causes membrane perturbations that result in internalization of the organisms .

Mikrobiologiia, 2002 Jan-Feb, 71(1), 37 - 48
{Comparative study of the elemental composition of vegetative and dormant microbial cells}; Muliukin AL et al.; X-ray microanalysis showed that vegetative cells, viable resting forms, and nonviable forms (micromummies) of the bacteria Bacillus cereus and Micrococcus luteus and the yeast Saccharomyces cerevisiae differ in the contents of bioelements S, P, Ca, and K and the Ca/K and P/S ratios . Viable resting forms (cystlike refractory cells and bacillar endospores) had more calcium and less phosphorus and potassium than vegetative cells, the difference being higher for bacilli than for micrococci and yeasts . The distinctive feature of all viable resting microbial forms was their low P/S ratios and high Ca/K ratios . The differences revealed in the cellular content and ratios of bioelements probably reflect changes in ionic homeostasis accompanying the transition of vegetative microbial cells to the dormant state . Relevant potassium parameters indicate that the membranes of viable resting forms retain their barrier function . At the same time, the nonviable forms, even morphologically intact, of B . cereus and S . cerevisiae exhibited an anomalously low content of potassium, while those of M . luteus had an anomalously high content of this element . This suggests that the cellular membranes of micromummies lose their barrier function, which results in a free diffusion of potassium ions across the membranes . The possibility of using the elemental composition parameters for quick analysis of the physiological state of microorganisms in natural environments is discussed.

Tunis Med, 2001 Oct, 79(10), 491 - 7
{Basis and guidelines for empirical antibody therapy in the management of community-acquired pneumonia}; Louzir B et al.; The choice of the antibiotic should be based on clinical, chest X-ray radiography and essentially microbiologic criteria . Incurrent practice treatment is more often empiric based on epidemiologic characteristics of the microbiologic agents and the particularities of each patient . A satisfactory approach requires, in addition, a perfect knowledge of different available antibiotics and the resistance of certain etiologic pathogens to these latters . because S . Pneumoniae is the most frequently encountered pathogen, B lactams and especially Penicillin G . and amoxicillin remain the most useful drugs prescribed for adults with risk factors . However, in advanced age patients and those with comorbidity, the spectrum should be enlarged and should include, besides S . Pneumoniae, H . influenzae and other Gram negative bacilli . When the pneumonia is more severe and has required hospitalization, the antimicrobial therapy must be immediate, multiple and large . The causal agent must be searched for desperately, so that the antimicrobial therapy can be adapted secondary to the results of the antibiogram.

Kekkaku, 2002 Feb, 77(2), 73 - 7
{A case of disseminated tuberculosis requiring extended period for the identification of Mycobacterium tuberculosis on culture}; Ashino J et al.; A 80-year-old male visited an outpatient department of a nearby hospital complaining of fever, cough, and poor appetite on June 2000 . The patient was diagnosed as bacterial pneumonia and was treated with antibiotics although specific cause could not be identified . After one month, he was hospitalized due to lack of improvement . After admission, acid-fast bacilli (AFB) was found from the bronchial washing . The patient was then transferred to our hospital . Upon admission, sputum smear examination was positive for AFB and MTB was confirmed by PCR . Therapy was initiated with INH 300 mg, RFP 450 mg, EB 1000 mg, and PZA 1000 mg, orally daily . However, on the day following the admission, he became unconscious . Brain MRI showed several small granulomas on the cortex of the bilateral anterior and temporal brain . Although AFB was not detected from the cerebrospinal fluid, tuberculous meningitis was suspected and steroid was given . Nine days after admission, the patient died due to tuberculous meningitis . The isolation of MTB had been attempted on Ogawa culture medium using patient's sputum and liquor, and it took 14 weeks to find colony growth both from sputum and liquor . In the autopsy, numerous granulomas were detected in his lung, liver, kidney, and pancreas . These findings indicate that disseminated growth of MTB occurred in vivo in spite of very slow growth of MTB in vitro.

Kekkaku, 2002 Feb, 77(2), 51 - 9
{Tuberculosis in the patients undergoing haemodialysis in Japan, 1996}; Sasaki Y et al.; We conducted a questionnaire survey on patients undergoing haemodialysis about the present situation of tuberculous incidence . They are immunocompromised hosts and are said to be at high risk of developing tuberculosis in many reports . (1) DESIGN Of the 167,192 patients on haemodialysis registered on December 31, 1996 in Japan, 71,411 patients were available for the questionnaire survey . Of the 2,893 hospitals used as the study subjects, 1,108 hospitals gave satisfactory replies . Of them, 141 hospitals reported that they had patients with tuberculosis in 1996, and 79 cases were collected by the detailed survey on tuberculosis patients conducted later . They included 45 male cases, 34 female cases for tuberculosis of all forms, 28 male cases, 15 female cases for pulmonary tuberculosis (PTB), 13 male cases, 4 female cases for tuberculosis bacilli positive pulmonary tuberculosis (TB positive PTB), and 17 male cases, 19 female cases for extrapulmonary tuberculosis . (2) RESULTS: In tuberculosis of all forms, the number of observed patients (O) against the number of patients expected (E) was calculated, and the standardized patients ratio (O/E ratio) was computed . It was 1.55 for male, 2.79 for female and 1.99 for total . The incidence of tuberculosis haemodialysis patients was significantly higher compared with the general population (p < 0.01) . As to PTB, the O/E ratio was 1.01 for male, 1.40 for female and 1.16 for total; the incidence of PTB was not significantly higher compared with the general population . With TB positive PTB, the O/E ratio was 0.96 for male, 0.80 for female and 0.97 for total, and no significant difference was found . As for extrapulmonary tuberculosis, the O/E ratio was 13.45 for male, 13.07 for female and 12.97 for total; the incidence of extrapulmonary tuberculosis in haemodialysis patients was significantly higher (p < 0.01), but it was lower than these reported in the past literature . The seventy nine cases consisted of 52 primary treatment cases, 23 retreatment cases, and 4 unknown cases . Out of 79 cases, 36 cases developed tuberculosis almost at the same time or within 1 year after undergoing haemodialysis, and thereafter it decreased gradually . Underlying diseases for haemodialysis were mainly glomerulonephritis and diabetic nephropathy . There were many patients who failed to notify to the public health centers after the diagnosis of tuberculosis was made, and it is needed to improve such a situation in the future . The prognosis of tuberculosis undergoing haemodialysis was poor . Three out of 43 patients with PTB and 2 out of 13 tuberculosis pleurisy cases died . (3) CONCLUSION: The risk of developing PTB in patients undergoing haemodialysis was not high compared with the general population, however, the risk was much higher for extrapulmonary tuberculosis . Moreover, the treatment outcome was not satisfactory in patients with PTB and pleurisy . As patients undergoing haemodialysis have the factors which suppress the cell-mediated immunity, it is required to restudy the measures to prevent development of tuberculosis, management and treatment in the future.

Res Microbiol, 2002 Mar, 153(2), 81 - 8
Spoligotyping and IS6110-RFLP typing of Mycobacterium tuberculosis from French Guiana: a comparison of results with international databases underlines interregional transmission from neighboring countries; Filliol I et al.; In this investigation, 94 clinical isolates of Mycobacterium tuberculosis obtained over a 3-year period (1996-1998) from French Guiana were characterized by spoligotyping and IS6110-RFLP and the patterns obtained were compared with genotypes representing the worldwide diversity in an international spoligotyping database (n = 4269) and a IS6110-RFLP database (n = 4189) . All the clustered isolates giving < or = 6 copies of IS6110 were further typed using the double-repetitive element (DRE)-PCR . The results obtained underlined the highly diverse nature of the M . tuberculosis population in French Guiana with potential links to neighboring countries within the Americas . It may be hypothesized that the genetic heterogeneity of tubercle bacilli in French Guiana is linked to the high number of imported cases of tuberculosis, that may account for as high as 68% of all tuberculosis cases . Although an epidemiological investigation based on direct interrogation of patients was not performed, available medical records suggested that the clustering of isolates was mostly linked to the following risk factors: pulmonary tuberculosis, smear-positive samples, foreign-born nationals and/or immigrants, and a high rate of HIV-TB coinfection . Thus the persisting foci of endemic disease and increased active transmission due to high population flux and HIV coinfection may be largely responsible for the relatively high incidence of tuberculosis in French Guiana.

Acta Cient Venez, 2001, 52 Suppl 1, 50 - 1
{Transcription regulation in the ribosomal RNA operon of mycobacteria}; Gonzalez Rico S et al.; Tuberculosis is a extremely important infectious disease, caused by the bacilli Mycobacterium tuberculosis . One of the characteristic of this bacteria is its very slow rate of growth, that allows it to survive for long periods of time inside the host cells . Among the genetic elements involved in growth regulation the operon rrn is of extreme importance . This operon contains the genes that code the three rRNA molecules, essential components of the bacterial ribosome . The tuberculosis bacilli, differently from most of the microorganisms, has a single copy of the rrn operon per genome, meaning that it must be submitted to very strict control mechanisms . Another important conclusion is that the sequences of the rrn operon constitute ideal targets for anti-mycobacterial drugs . In this work we have studied some of the elements involved in transcription control in M . tuberculosis, particularly those present in the leader region of the operon . By using basic molecular biology techniques we have identified sequence elements in the leader region that seem to be involved in the control of transcription elongation, by a mechanism related to anti-termination.

Acta Cient Venez, 2001, 52 Suppl 1, 36 - 9
Resistance to antimicrobial agents in Mycobacterium tuberculosis; Pfyffer GE; According to the World Health Organization (WHO) more people will currently die of tuberculosis (TB) than in any other year in history . Of equal concern are the emergence and nosocomial transmission of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis . Only recently, with the advent of new molecular biological techniques, the mechanisms of drug resistance in TB bacilli are more and more understood . In M . tuberculosis, the primary mechanism of drug resistance seems to be exclusively confined to chromosomal DNA and not, as in other bacteria, to mobile genetic elements as well.

Folia Microbiol (Praha), 2001, 46(5), 363 - 70
Nontraditional microbial bioactive metabolites; Behal V; Microorganisms produce low-molar-mass secondary metabolites exhibiting different biological activities, which are used . e.g., in medicine as antimicrobial and antifungal agents, alkaloids and toxins . Some of these substances have highly diverse biological activities and unusual structures . They are produced by streptomycetes, fungi, and bacilli, but interesting products have also been obtained from microorganisms growing in extreme conditions . Several thousands of microbial products have so far been discovered and many other, which can be potentially useful and/or prospective for human use, can still be in the offing.

Am J Respir Crit Care Med, 2002 Mar 15, 165(6), 812 - 7
Limited transmission of multidrug-resistant tuberculosis despite a high proportion of infectious cases in Los Angeles County, California; Nitta AT et al.; Preventing transmission of multidrug-resistant tuberculosis is critical because of treatment toxicity, cost, and the lack of effective therapy for latent infection . We attempted to determine the extent of transmission in Los Angeles County by comparing relatedness of multidrug-resistant tuberculosis cases using restriction fragment length polymorphism and by cross-matching contact information to the Tuberculosis Registry . Strain typing was done on isolates of 102 pulmonary multidrug-resistant cases identified between August 1993 and 1998 . Seventy-one (70%) of the cases had cavitary lesions on chest radiograph, and 94 (92%) had sputa smear-positive for acid fast bacilli . Fifteen (15%) of the cases were known to be infected with human immunodeficiency virus . Four molecular clusters of two cases each and one closely related pair were identified among the 102 cases; contact investigation successfully identified all clusters but one . Among 946 contacts identified and cross-matched with the county's Tuberculosis Registry, one secondary case due to drug-resistant Mycobacterium bovis was found . To summarize, a very high proportion of pulmonary multidrug-resistant tuberculosis cases in Los Angeles County were infectious . Molecular strain typing indicated limited spread of disease, although it underestimated transmission compared with contact investigation . We believe aggressive surveillance and case management were critical to limiting the spread of multidrug- resistant tuberculosis.

Proc Natl Acad Sci U S A, 2002 Mar 19, 99(6), 3684 - 9 Epub 2002 Mar 12.
A new evolutionary scenario for the Mycobacterium tuberculosis complex; Brosch R et al.; The distribution of 20 variable regions resulting from insertion-deletion events in the genomes of the tubercle bacilli has been evaluated in a total of 100 strains of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canettii, Mycobacterium microti, and Mycobacterium bovis . This approach showed that the majority of these polymorphisms did not occur independently in the different strains of the M . tuberculosis complex but, rather, resulted from ancient, irreversible genetic events in common progenitor strains . Based on the presence or absence of an M . tuberculosis specific deletion (TbD1), M . tuberculosis strains can be divided into ancestral and "modern" strains, the latter comprising representatives of major epidemics like the Beijing, Haarlem, and African M . tuberculosis clusters . Furthermore, successive loss of DNA, reflected by region of difference 9 and other subsequent deletions, was identified for an evolutionary lineage represented by M . africanum, M . microti, and M . bovis that diverged from the progenitor of the present M . tuberculosis strains before TbD1 occurred . These findings contradict the often-presented hypothesis that M . tuberculosis, the etiological agent of human tuberculosis evolved from M . bovis, the agent of bovine disease . M . canettii and ancestral M . tuberculosis strains lack none of these deleted regions, and, therefore, seem to be direct descendants of tubercle bacilli that existed before the M . africanum-->M . bovis lineage separated from the M . tuberculosis lineage . This observation suggests that the common ancestor of the tubercle bacilli resembled M . tuberculosis or M . canettii and could well have been a human pathogen already.

Ann Rheum Dis, 2001 Nov, 60 Suppl 3, iii6 - 12
Differential roles of Toll-like receptors in the elicitation of proinflammatory responses by macrophages; Jones BW et al.; BACKGROUND: Mammalian Toll-like receptor (TLR) proteins are pattern recognition receptors for a diverse array of bacterial and viral products . Gram negative bacterial lipopolysaccharide (LPS) activates cells through TLR4, whereas the mycobacterial cell wall glycolipids, lipoarabinomannan (LAM) and mannosylated phosphatidylinositol (PIM), activate cells through TLR2 . Furthermore, short term culture filtrates of M . tuberculosis bacilli contain a TLR2 agonist activity, termed soluble tuberculosis factor (STF), that appears to be PIM . It was recently shown that stimulation of RAW264.7 murine macrophages by LPS, LAM, STF, and PIM rapidly activated NF-kappaB, AP1, and MAP kinases . RESULTS: This study shows that signalling by TLR2 and TLR4 also activates the protein kinase Akt, a downstream target of phosphatidylinositol-3'-kinase (PI-3-K) . This finding suggests that activation of PI-3-K represents an additional signalling pathway induced by engagement of TLR2 and TLR4 . Subsequently, the functional responses induced by the different TLR agonists were compared . LPS, the mycobacterial glycolipids, and the OspC lipoprotein (a TLR2 agonist) all induced macrophages to secrete tumour necrosis factor alpha (TNFalpha), whereas only LPS could induce nitric oxide (NO) secretion . Human alveolar macrophages also exhibited a distinct pattern of cellular response after stimulation with TLR2 and TLR4 agonists . Specifically, LPS induced TNFalpha, MIP-1beta, and RANTES production in these cells, whereas the TLR2 agonists induced only MIP-1beta production . CONCLUSION: Together, these data show that different TLR proteins mediate the activation of distinct cellular responses, despite their shared ability to activate NF-kappaB, AP1, MAP kinases, and PI-3-K.

Chest, 2002 Mar, 121(3), 774 - 81
Adequately washed bronchoscope does not induce false-positive amplification tests on bronchial aspirates in the diagnosis of pulmonary tuberculosis; Shim TS et al.; STUDY OBJECTIVES: To investigate the clinical usefulness of amplification (COBAS AMPLICOR; Roche Diagnostics Systems; Branchburg, NJ) on bronchoscopic aspirate specimens in the diagnosis of pulmonary tuberculosis, with particular regard to the possibility of false-positive results in subsequent specimens due to residual Mycobacterium tuberculosis DNA . DESIGN AND SETTING: A prospective clinical study at a tertiary referral medical center . PARTICIPANTS AND METHODS: Four hundred fiberoptic bronchoscopic procedures were performed, using seven bronchoscopes on 335 consecutive patients, for therapeutic or diagnostic purposes . Serial bronchial aspirates were collected and tested for M tuberculosis, using COBAS AMPLICOR (CA) . Bronchoscopes were cleaned and disinfected automatically, between patient use, by the same endoscope washer . The name of each bronchoscope and the sequence of its use were recorded, together with the sequence of washing . The CA results were compared with the bacteriologic and histologic results for M tuberculosis infection . When there was a suspicion of contamination, outward polymerase chain reaction analysis was performed . RESULTS: Of 392 specimens (332 subjects), excluding the 8 specimens (4 subjects) in which bacteriologic and histologic analyses were omitted, a smear-positive result for acid-fast bacilli (AFB), culture-positive or biopsy-positive results, and CA-positive results were obtained in 16, 49, and 32 specimens, respectively . In AFB smear-positive subjects, the sensitivity, specificity, positive predictive values (PPVs), and negative predictive values (NPVs) were 92%, 67%, 92%, and 67%, respectively . In AFB smear-negative subjects, the sensitivity, specificity, PPV, and NPV values were 38%, 99%, 74%, and 94%, respectively . The CA test was more sensitive than the AFB smears for the diagnosis of pulmonary tuberculosis (53% vs 27%, respectively; p < 0.05) . False-positive CA results were seen in only six specimens . Three of these six subjects received a diagnosis of pulmonary tuberculosis on clinical and radiologic grounds, and none of the six results seemed to be associated with bronchoscopic cross-contamination . CONCLUSIONS: Adequately cleaned and disinfected bronchoscopes did not cause false-positive amplification test results for M tuberculosis on bronchial aspirates by cross-contamination . Furthermore, sensitivity was greater with the CA tests . Therefore, CA tests on bronchial aspirates seem to be useful in the diagnosis of pulmonary tuberculosis.

Med Sci Monit, 2002 Mar, 8(3), CS26 - 9
Cat-scratch disease in an immunocompromised host; Harsch IA et al.; BACKGROUND: The main causative agents of cat-scratch disease are Bartonella henselae, tiny, gram-negative bacilli . The disease usually has a benign course with the development of a papule at the inoculation site, followed by regional lymphadenopathy . In most cases, complete resolution occurs, but in immunocompromised hosts, the course of the disease can be aggravated . CASE REPORT: A patient received methotrexate and corticosteroids for 3 months due to rheumatoid arthritis . He developed fever, exanthema and leukopenia under methotrexate therapy . Dark red indurations with central ulcerations at his right thigh revealed a further problem apart from the methotrexate-induced leucopenia and immunosuppression . The ulcerations were the remainders of recurrent scratches from the patient's cat . The patient's antibody titers against Bartonella henselae remained low and inguinal lymph node swelling was only for a short time to be observed, this reaction obviously weakened as a result of the immunosuppression . However, the typical course, the exclusion of other reasons for the exanthema and the rapid improvement of the patient's condition after antibiotic treatment ascertained the diagnosis . CONCLUSIONS: In immunocompromised hosts, diseases with a typically benign course can become severe and life-threatening illnesses . Ownership of pets should be taken into consideration before onset of an immunosuppressive therapy.

Bull World Health Organ, 2002, 80(1), 16 - 25
Etiology of child mortality in Goroka, Papua New Guinea: a prospective two-year study; Duke T et al.; OBJECTIVE: To collect accurate data on disease- and microbial-specific causes and avoidable factors in child deaths in a developing country . METHODS: A systematic prospective audit of deaths of children seen at Goroka Hospital in the highlands of Papua New Guinea was carried out . Over a 24-month period, we studied 353 consecutive deaths of children: 126 neonates, 186 children aged 1-59 months, and 41 children aged 5-12 years . FINDINGS: The most frequent age-specific clinical diagnoses were as follows: for neonates--very low birth weight, septicaemia, birth asphyxia and congenital syphilis; for children aged 1-59 months--pneumonia, septicaemia, marasmus and meningitis; and for children aged 5-12 years--malignancies and septicaemia . At least one microbial cause of death was identified for 179 (50.7%) children and two or more were identified for 37 (10.5%) . Nine microbial pathogens accounted for 41% of all childhood deaths and 76% of all deaths that had any infective component . Potentially avoidable factors were identified for 177 (50%) of deaths . The most frequently occurring factors were as follows: no antenatal care in high-risk pregnancies (8.8% of all deaths), very delayed presentation (7.9%), vaccine-preventable diseases (7.9%), informal adoption or child abandonment leading to severe malnutrition (5.7%), and lack of screening for maternal syphilis (5.4%) . Sepsis due to enteric Gram-negative bacilli occurred in 87 (24.6%) . The strongest associations with death from Gram- negative sepsis were adoption/abandonment leading to severe malnutrition, village births, and prolonged hospital stay . CONCLUSIONS: Reductions in child mortality will depend on addressing the commonest causes of death, which include disease states, microbial pathogens, adverse social circumstances and health service failures . Systematic mortality audits in selected regions where child mortality is high may be useful for setting priorities, estimating the potential benefit of specific and non-specific interventions, and providing continuous feedback on the quality of care provided and the outcome of health reforms.

Clin Nutr, 2001 Dec, 20(6), 527 - 34
Colonization and bacteremia risk factors in parenteral nutrition catheterization; Llop J et al.; AIMS: 1) To establish the relationship between the kind of microorganism that colonizes parenteral nutrition catheters and several risk factors related to catheterization and patient characteristics . 2) To investigate the risk factors associated to bacteremia episodes originated in these colonized catheters . METHOD: An observational, non-controlled, retrospective and cohorts study of the parenteral nutrition catheters implanted between 1988 and 1994 in our hospital . Risk factors were studied in 6 multiple-logistic regression models . RESULTS: 3632 catheters were studied . Incidences of colonization and bacteremia per 1000 days of catheterization were 17.56 and 3.93, respectively . Coagulase-negative staphyloccoci (CNS) were the most frequently isolated microorganisms . The colonization risk factors were: insertion site for all the microorganisms except fungi, catheterization time for CNS and fungi, hospitalization area, sex and age for CNS model, the existence of other infectious foci for Gram negative bacilli (GNB), S . aureus and other microorganisms, hypoalbuminemia for GNB model, and neoplasm for other microorganisms . The bacteremia risk factors were jugular insertion site, catheterization time greater than 10 days, catheter's hub colonization, and catheter colonization by gram-negative bacilli, fungi and S . aureus . CONCLUSION: Risk factors for catheter colonization vary depending on the microorganism which colonizes the catheter .

Indian J Exp Biol, 2001 Oct, 39(10), 984 - 8
A dot-immunobinding assay (dot-Iba) for rapid diagnosis of pulmonary tuberculosis; Sumi MG et al.; IgG antibody to Mycobacterium tuberculosis from the sera of patients with 'definite' pulmonary tuberculosis (PT) was isolated and coupled with Cyanogen bromide-Sepharose 4B . Using an immunoabsorbent affinity chromatography, 14 kDa antigen was recovered from the culture filtrates of M . tuberculosis . With this mycobacterial antigen, a dot immunobinding assay (Dot-Iba) was developed for the detection of specific antibody to M . tuberculosis in the sera of patients with PT and controls . The assay gave positive results in all the 12 sputum-smear positive {acid fast bacilli (AFB)} patients with PT and gave negative results in the 50 sera from control groups . The Dot-Iba as described in this study, is simple, rapid and specific for laboratory diagnosis of PT.

Indian J Exp Biol, 2001 Oct, 39(10), 962 - 83
Leprosy bacillus--possibly the first chemoautotrophic human pathogen cultivated in vitro and characterised; Chakrabarty AN et al.; Leprosy bacillus (LB) and leprosy derived in vitro culture forms, the chemoautotrophic nocardioform (CAN) bacteria, showed an extremely close homology and identity with each other as regards a chemoautotrophic nutritional pattern, a nocardioform morphology, a weak acid-fastness coupled with Gram and Gomori's stain positivity, an exclusive mycolate and lipid profile, a phenolic glycolipid (PGL-I) and a highly sequestrated DNA characteristic, namely, a unique small size, a low G+C % mole, an exceptionally high gamma and UV radiation resistance, and a high thermal resistance . LB/CAN bacteria (CANb) gave positive signals for 36 kDa protein PCR, as well as, for 65 kDa epitope, and hybridisation with two or more probes and also by RFLP-analysis . Both LB/and CAN bacteria exhibited bacillary multiplication in the mouse footpads (MFP), nerve infiltration and evidences for local pathogenicity associated with pronounced systemic invasion . A highly reproducible mutilation model could be established which enabled a successful application of the postulates of Koch . The proof of their total identity was their anergic reactions in LL cases counterpoised against Mitsuda type strong nodular responses, mirroring the reactions of leprosy bacilli in TT cases, in accordance with the dictum of XIth International Leprosy Congress (1978) . Thus, the chemoautotrophic nutritional requirements of LB, entirely unsuspected for a medically important pathogenic bacterium, having dimorphic (both bacillary and mycelial) characters with spores, mycelia and granules and unique pathogenicity of multilation manifested through the virulence factor, the enzyme collagenase, made LB or M leprae the highly enigmatic bacterium for so long.

Indian J Med Sci, 2001 Jul, 55(7), 382 - 92
Drug resistance among tubercle bacilli from pulmonary tuberculosis cases in central India; Hemvani N et al.; An estimate of drug resistance is extremely important in the epidemiology and control of tuberculosis . Data on drug resistance among mycobacterial isolates from sputum samples analysed at Microbiology dept . of Choithram Hospital and Research Centre, Indore, M.P . is presented here . Drug sensitivity testing was carried out on 1426 Mycobacterial isolates by the method of proportion using critical concentration in Lowenstein Jensen medium . Resistance for Isoniazid, streptomycin, and pyrazinamide was found to be high (54.2%, 41.5% and 50% respectively) and was followed by resistance to rifampin (25%) and ethambutol (22%) . Resistance for kanamycin, p-aminosalicylic acid, thiacetazone and ciprofloxacin was much lower (18%, 13%, 6.5% and 3.6% respectively) . Only 12% of the isolates were sensitive to all the anti-TB drugs while resistance to two, three, and four or more drugs was in the range of 20-25% . Pattern wise, simultaneous resistance to INF and Rifampin with or without resistance to other drugs was observed in 8.1% while resistance for Isoniazid + pyrazinamide and Isoniazid + streptomycin was 11.9 and 11.5% respectively . Resistance for Isoniazid + ethambutol was the lowest (5.1%) . Growing multiple drug resistance among tubercle bacilli warrant urgent attention in tuberculosis control programme.

Pathol Int, 2001 Oct, 51(10), 807 - 11
Malignant lymphoma of bronchus-associated lymphoid tissue (BALT) coexistent with pulmonary tuberculosis; Inadome Y et al.; A case in which malignant lymphoma occurred in association with a tuberculosis focus in a 70-year-old man is reported . Surrounding the epithelioid cell granulomas with caseous necrosis was a dense and diffuse monotonous infiltration of atypical lymphoid cells . Acid-fast bacilli were found in the granulomas and pulmonary tuberculosis was diagnosed . The infiltrating atypical lymphoid cells occasionally invaded the respiratory epithelium producing lymphoepithelial lesions . Immunohistochemically, the lymphoid cells were positive for CD20, and clonal rearrangement of the immunoglobulin heavy chain gene was demonstrated by polymerase chain reaction (PCR) . We diagnosed the lesion as a pulmonary malignant lymphoma of bronchus-associated lymphoid tissue (BALT) occurring in the background of tuberculosis . This is the first reported case of pulmonary BALT lymphoma coexistent with pulmonary tuberculosis.

Int J Lepr Other Mycobact Dis, 2001 Sep, 69(3), 177 - 86
Distinct histopathological patterns in single lesion leprosy patients treated with single dose therapy (ROM) in the Brazilian Multicentric Study; Costa MB et al.; This paper aims to describe the histomorphologic features of skin biopsies of single lesion leprosy patients recruited at outpatient clinics in four Brazilian states in the Northeast (Amazonas and Rondonia), Southeast (Rio de Janeiro) and Center-West (Goias) between October 1997 and December 1998 . Patients clinically diagnosed as single skin lesion paucibacillary (SSL-PB) leprosy had a standard 4-mm punch biopsy taken from the lesion before rifampin, ofloxacin, minocycline (ROM) therapy . The features of the cellular inflammatory infiltrates, the presence of nerve involvement and acid-fast bacilli (AFB) were used to categorize SSL-PB biopsies into different histopathological groups . Two-hundred-seventy-eight (93.0%) out of 299 patients had a skin biopsy available . Seven single lesion patients were diagnosed as BL or LL leprosy types (MB) by the histopathological exams and 12 cases were excluded due to other skin diseases . Therefore, 259 patients had skin lesions with histomorphological features compatible with PB leprosy categorized as follows: 33.6% (N = 87) of the biopsies represented well-circumscribed epithelioid cell granuloma (Group 1); 21.6% (N = 56) less-circumscribed epithelioid cell granuloma (Group 2); 12.0% (N = 31) were described as mononuclear inflammatory infiltrate permeated with epithelioid cells (Group 3), and 29.7% (N = 77) had perivascular/periadnexal mononuclear inflammatory infiltrate (Group 4) . Minimal/no morphological alteration in the skin was detected in only 8 (3.1%) SSL-PB patients categorized as Group 5, who were considered to have leprosy by clinical parameters . SSL-PB leprosy patients recruited in a multicentric study presented histomorphology readings comprising the whole PB leprosy spectrum but also a few MB cases . These results indicate heterogeneity among SSL-PB patients, with a predominance of well-circumscribed and less-circumscribed epithelioid cell granulomas (Groups 1 and 2) in the sites studied and the heterogeneity of local cellular immune response.

Br J Cancer, 2002 Feb 1, 86(3), 336 - 41
Clinical and immunological assessment of Mycobacterium vaccae (SRL172) with chemotherapy in patients with malignant mesothelioma; Mendes R et al.; The objectives of this study were to determine the toxicity of intratumoural/intrapleural SRL172 in addition to intradermal SRL172 and standard chemotherapy (mitomycin-C, vinblastine and cisplatin) in patients with malignant mesothelioma . Patients received chemotherapy (mitomycin-C: 8 mg m(-2), vinblastine: 6 mg m(-2), cisplatin 50 mg m(-2)) on a 3-weekly basis for up to six courses . IP SRL172 injections were given 3-weekly prior to chemotherapy and escalated in groups of three patients from 1 microg to 1 mg bacilli in 10-fold increments . Patients were also given ID SRL172 at a dose of 1 mg bacilli 4-weekly . Patients were assessed for toxicity after each course of chemotherapy and for response by CT imaging . Immuno-haematological parameters were analyzed pre-treatment and 1 month after completion of treatment . There was no dose limiting toxicity with IP SRL172 although there was greater toxicity at the highest dose (n=13) . There were six out of 16 partial responses (37.5%) . Haemato-immunological parameters, measured in seven patients pre and post-therapy, revealed that response rate correlated with a decrease in platelet count and there was an increase in activation of natural killer cells and a decrease in the percentage of IL-4 producing T cells in all tested patients post-treatment . SRL172 can be given safely into tumour deposits and the pleural cavity in patients with malignant mesothelioma and we have established the dose for phase II testing .

Lett Appl Microbiol, 2002, 34(3), 194 - 8
The occurrence and properties of Bacillus thuringiensis isolated from free-living animals; Swiecicka I et al.; AIMS: To assess the prevalence and properties of Bacillus thuringiensis isolated from the intestines of small mammals . METHODS AND RESULTS: Bacillus thuringiensis was found in 11% of rodents and 17% of insectivores . Using PFGE of chromosomal DNA, SDS-PAGE of whole-cell proteins and biochemical tests (API system), 12 isolates and three reference strains were classified . Numerical analysis revealed 61% and 89% similarity of protein profiles and biochemical properties of the bacilli, respectively . The results of PFGE were consistent with the outcomes of the analysis of protein profiles . CONCLUSIONS: Although B . thuringiensis is not common in the intestines of small mammals, it is heterogeneous at the genotypic and phenotypic level . SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented here help to explain the diversity and the ecological significance of B . thuringiensis . Future study should focus on the toxic activity of the isolated strains.

J Microbiol Methods, 2002 May, 49(3), 295 - 300
Reliability of hsp65-RFLP analysis for identification of Mycobacterium species in cultured strains and clinical specimens; de Magalhaes VD et al.; The purpose of this study was to evaluate the reliability of an amplification restriction analysis based method (hsp65-RFLP) to detect and identify mycobacterial species in clinical samples and cultures with low number of bacilli . We examined 247 clinical specimens and 88 culture vials, comparing hsp65-RFLP results with conventional culture/biochemical tests . The analytical sensitivity of the method was assessed with cerebrospinal fluid (CSF), broncho-alveolar lavage (BAL), sputum, water, and 12B medium containing defined amounts of mycobacterial chromosome . We detected the equivalent of 10(3) cells per ml in all samples, except sputum, the most common source of clinical sample for mycobacterial testing, which presented inhibition throughout . We investigated two purification procedures to overcome inhibition of DNA amplification: DNAzol and phenol/chloroform . The former was superior, eliminating inhibition in 93.7% of the clinical samples . The technique was effective for bacterial cultures, including those with very low growth indices (GIs), substantially abbreviating time for diagnosis, but showed low sensitivity (25%) when applied to clinical samples, an issue that has never been extensively assessed by other researchers.

J Feline Med Surg, 2002 Mar, 4(1), 43 - 59
Feline leprosy: two different clinical syndromes; Malik R et al.; Feline leprosy refers to a condition in which cats develop granulomas of the subcutis and skin in association with intracellular acid-fast bacilli that do not grow on routine laboratory media . In this study, the definition was extended to include cases not cultured, but in which the polymerase chain reaction (PCR) identified amplicons characteristic of mycobacteria . Tissue specimens from 13 such cases from eastern Australia were obtained between 1988 and 2000 . This cohort of cats could be divided into two groups on the basis of the patients' age, histology of lesions, clinical course and the sequence of 16S rRNA PCR amplicons.One group consisted of four young cats (less than 4 years) which initially developed localised nodular disease affecting the limbs . Lesions progressed rapidly and sometimes ulcerated . Sparse to moderate numbers of acid-fast bacilli were identified using cytology and/or histology, typically in areas of caseous necrosis and surrounded by pyogranulomatous inflammation . Organisms did not stain with haematoxylin and ranged from 2 to 6 microm (usually 2 to 4 microm) . Mycobacterium lepraemurium was diagnosed in two cases based on the sequence of a 446 bp fragment encompassing the V2 and V3 hypervariable regions of the 16S rRNA gene a different sequence was obtained from one additional case, while no PCR product could be obtained from the remaining case . The clinical course was considered aggressive, with a tendency towards local spread, recurrence following surgery and development of widespread lesions over several weeks . The cats resided in suburban or rural environments . A second group consisted of nine old cats (greater than 9 years) with generalised skin involvement, multibacillary histology and a slowly progressive clinical course . Seven cats initially had localised disease which subsequently became widespread, while two cats allegedly had generalised disease from the outset . Disease progression was protracted (compared to the first group of cats), typically taking months to years, and skin nodules did not ulcerate . Microscopically, lesions consisted of sheets of epithelioid cells containing large to enormous numbers of acid-fast bacilli 2 to 8 microm (mostly 4 to 6 microm) which stained also with haematoxylin . A single unique sequence spanning a 557 bp fragment of the 16S rRNA gene was identified in six of seven cases in which it was attempted . Formalin-fixed paraffin-embedded material was utilised by one laboratory, while fresh tissue was used in another . The same unique sequence was identified despite the use of different primers and PCR methodologies in the two laboratories . A very slow, pure growth of a mycobacteria species was observed on Lowenstein-Jensen medium (supplemented with iron) and semi-solid agar in one of three cases in which culture was attempted at a reference laboratory . Affected cats were domicile in rural or semi-rural environments . These infections could generally be cured using two or three of rifampicin (10-15 mg/kg once a day), clofazimine (25 to 50 mg once a day or 50 mg every other day) and clarithromycin (62.5 mg per cat every 12 h).These findings suggest that feline leprosy comprises two different clinical syndromes, one tending to occur in young cats and caused typically by M lepraemurium and another in old cats caused by a single novel mycobacterial species .

Zhonghua Bing Li Xue Za Zhi, 2001 Apr, 30(2), 89 - 92
{Mycobacterial spindle cell pseudotumor of lymph nodes}; Yin H et al.; OBJECTIVE: To study the clinicopathological of mycobacterial spindle cell pseudotumor in lymph nodes . METHODS: Two cases of spindle cell pseudotumor in the lymph nodes of infants were examined using light microscopy . The sections were stained with HE, Ziehl-Neelsen and studied immunohistochemically . RESULTS: Both infants were 1 year of age and had a history of Bacilli Calmette Guerin (BCG) vaccination before discovery of their lymph node lesions . Clinically, the patients presented fever and enlarged lymph nodes . Histopathologically, the lesions showed proliferation of spindle cells arranged in bundles, storiform patterns with abundant capillaries and lymphocyte, plasma cell and neutrophil infiltration forming a spindle cell pseudotumor . Most of the spindle cells were phagocytic cells which contained large amounts of mycobactrium as observed by Ziehl-Neelsen staining . Immunohistochemical staining showed strongly positive result for Mac387 and lysosome, but negative for S-100 in spindle cells and histocytes . CONCLUSIONS: Mycobacterial spindle cell pseudotumor of lymph nodes may occur not only in immunodeficient patients (such as AIDS) but also in infants after BCG vaccination . The lesions may mimic spindle cell tumor in lymph nodes, therefore it is very important to recognize these lesions in order to treat the patient properly.

J Infect Dis, 2002 Feb 15, 185 Suppl 1, S1 - 8
The study of Mycobacterium leprae infection in interferon-gamma gene--disrupted mice as a model to explore the immunopathologic spectrum of leprosy; Adams LB et al.; Mycobacterium leprae infection was evaluated in interferon-gamma knockout (GKO) mice . At 4 months, growth of the bacilli in the footpads of GKO mice plateaued a log(10) higher than that in control mice . Control mice exhibited mild lymphocytic and histiocytic infiltrates, whereas GKO mice developed large, unorganized infiltrates of epithelioid macrophages and scattered CD4 and CD8 T cells . Flow cytometric analysis of popliteal lymph node cells demonstrated similar profiles of T cells; however, GKO cells exhibited an elevated proliferative response to M . leprae antigen . Expression of inducible nitric oxide synthase mRNA was decreased in GKO mice, whereas macrophage inflammatory protein-1alpha and interleukin-4 and -10 mRNA expression were augmented . Control and GKO activated macrophages inhibited bacterial metabolism and produced nitrite . Thus, although deficient in an important Th1 cytokine, GKO mice possess compensatory mechanisms to control M . leprae growth and feature elements resembling mid-borderline leprosy in humans.

J Med Microbiol, 2002 Feb, 51(2), 131 - 7
Pulmonary granulomas of guinea pigs induced by inhalation exposure of heat-treated BCG Pasteur, purified trehalose dimycolate and methyl ketomycolate; Sugawara I et al.; This study was designed to determine the identity of granulomatogenic substances in Mycobacterium bovis BCG Pasteur . When heat-treated BCG Pasteur bacilli were introduced into the lungs of guinea-pigs by an inhalation exposure apparatus, pulmonary granulomas without necrosis developed . Furthermore, when four kinds of mycolates derived from M . tuberculosis Aoyama B strain were introduced into the lungs by the same method, only trehalose 6,6'-dimycolate (TDM) and methyl ketomycolate induced pulmonary granulomas without central necrosis . The pulmonary granulomas consisted of epithelioid macrophages and lymphocytes . When a mixture of TDM and anti-TDM antibody was introduced into the lungs, development of granulomatous lesions was reduced . These data indicate that TDM and methyl ketomycolate are potent granulomatogenic reagents.

BMC Microbiol . 2002;2(1):2 . Epub 2002 Jan 30.
Killing of Mycobacterium avium subspecies paratuberculosis within macrophages; Bannantine JP et al.; BACKGROUND: Mycobacterium avium subspecies paratuberculosis (M . paratuberculosis) is a facultative intracellular pathogen that resides within host macrophages during infection of ruminant animals . We examined survival of M . paratuberculosis infections within cultured macrophages to better understand the interplay between bacterium and host . RESULTS: Serial plating of M . paratuberculosis infected macrophage lysates on Herold's egg yolk medium showed that mycobacterial replication takes place between 0 and 24 hours post-infection . This initial growth phase was followed by a steady decline in viability over the next six days . Antibodies against M . paratuberculosis were affinity purified and used in conjunction with transmission electron microscopy to track the development of intracellular bacilli . Immunogold labeling of infected macrophages with antibody against M . paratuberculosis showed degraded intracellular mycobacteria that were unrecognizable by morphology alone . Conversely, when macrophages were heavily infected with M . paratuberculosis, no degraded forms were observed and macrophages were killed . CONCLUSIONS: We present a general description of M . paratuberculosis survival within cultured macrophages using transmission electron microscopy and viability counts . The results of this study provides further insight surrounding M . paratuberculosis-macrophage infections and have implications in the pathogenesis of M . paratuberculosis, a pathogen known to persist inside cattle for many years.

Rev Med Interne, 2002 Jan, 23(1), 4 - 8
{Oropharyngeal colonization by Gram-negative bacteria in elderly hospitalized patients: incidence and risk factors}; Hiar I et al.; PURPOSE: In a prospective study, we determined the predisposing conditions and the course of aerobic Gram-negative bacilli oropharyngeal colonization during the hospital stay in patients older than 70 years . METHODS: We studied 116 patients admitted in our internal medicine unit . Medical characteristics were documented for each patient . Oropharyngeal swabs were collected on the first, third, and seventh days after admission, and then once a week until discharge . Positive culture was defined as the presence of at least one colony of Gram-negative bacilli . RESULTS: We collected 306 samples from the 116 patients . The mean hospital stay was 10.6 days . Fifty patients (43%) had at least one positive oropharyngeal culture . Thirty-three patients (30%) were colonized on admission, but this prevalence decreased during hospital stay: 20% on the third day and 14% on the seventh day . In an univariate analysis, prior administration of antibiotics (P < 0.01) and use of nonsteroid anti-inflammatory drugs (P < 0.05) were associated with colonization . CONCLUSION: Aerobic Gram-negative bacilli oropharyngeal colonization was transient during the hospital stay in the elderly . The use of nonsteroidal anti-inflammatory drugs and the previous treatment with antibiotics are possible risk factors for colonization.

Clin Microbiol Infect, 1999 Jun, 5(6), 319 - 326
Evaluation of the Rapid ID 32A system for identification of anaerobic Gram-negative bacilli, excluding the Bacteroides fragilis group; Downes J et al.; OBJECTIVE: To evaluate the Rapid ID 32A system (bioMerieux, Marcy-l'Etoile, France) for the identification of anaerobic Gram-negative bacilli, excluding the Bacteroides fragilis group . METHODS: Five hundred and twenty-eight identified clinical isolates of non-B . fragilis group anaerobic Gram-negative bacilli were tested in the Rapid ID 32A system, and identifications were compared with those obtained with conventional biochemical tests and gas-liquid chromatography . RESULTS: The Rapid ID 32A system correctly identified 280 (60.9%) of the 460 isolates tested for which taxa were included in the database, without the need for additional testing . A further 97 (21.1%) isolates were correctly identified to species level following the performance of complementary tests recommended by the manufacturer . Fifty-nine (12.8%) isolates were identified at the genus level only, and 21 (4.6%) were misidentified at the species level . Three isolates of Prevotella were not identified by the system . Of the 68 isolates belonging to taxa not included in the database, no identification was obtained for 33 (48.5%), while 35 (51.5%) were misidentified . CONCLUSIONS: The Rapid ID 32A system provided a rapid and reliable method for the identification of non-B . fragilis group, anaerobic Gram-negative bacilli to the genus level, while the success of species-level identification varied with different taxa . There was poor discrimination between Fusobacterium nucleatum and F . necrophorum, between Porphyromonas asaccharolytica and Porphyromonas endodontalis, and between Prevotella buccalis, Prevotella denticola, Prevotella loescheii, Prevotella melaninogenica and Prevotella oralis . The need to perform conventional complementary tests on 149 (32.4%) of the 460 isolates compromised the usefulness of the system for rapid species identification.

Nihon Kokyuki Gakkai Zasshi, 2001 Nov, 39(11), 857 - 61
{An adult case of mediastinal tuberculous lymphadenitis}; Kanaji N et al.; A 40-year-old woman was admitted to the hospital with general fatigue and cough . Chest CT films revealed mediastinal lymphadenopathy with multiple low density areas, but no pulmonary lesions . There were no abnormal findings on neck, abdominal or pelvic CT . A PPD skin test was strongly positive, but M . tuberculosis bacilli were not found in the sputum . Mediastinal tuberculous lymphadenitis was diagnosed histologically and bacteriologically from specimens obtained by mediastinoscopy . Fiberoptic bronchoscopy did not reveal tracheobronchial tuberculosis . Follow-up chest CT three months after the start of antituberculosis chemotherapy with isoniazid, rifampicin and ethambutol hydrochloride showed that the mediastinal lymph nodes were decreased in size . Mediastinal tuberculous lymphadenitis in adults is rare, but the number of reports has increased . Mediastinal tuberculous lymphadenitis in adults must be distinguished from other causes of mediastinal masses . In this case, mediastinoscopy was very useful for differential diagnosis.

Infect Immun, 2002 Mar, 70(3), 1501 - 6
Survival of Tropheryma whipplei, the agent of Whipple's disease, requires phagosome acidification; Ghigo E et al.; Tropheryma whipplei was established as the agent of Whipple's disease in 2000, but the mechanisms by which it survives within host cells are still unknown . We show here that T . whipplei survives within HeLa cells by controlling the biogenesis of its phagosome . Indeed, T . whipplei colocalized with lysosome-associated membrane protein 1, a membrane marker of late endosomal and lysosomal compartments, but not with cathepsin D, a lysosomal hydrolase . This defect in phagosome maturation is specific to live organisms, since heat-killed bacilli colocalized with cathepsin D . In addition, T . whipplei survived within HeLa cells by adapting to acidic pH . The vacuoles containing T . whipplei were acidic (pH 4.7 +/- 0.3) and acquired vacuolar ATPase, responsible for the acidic pH of late phagosomes . The treatment of HeLa cells with pH-neutralizing reagents, such as ammonium chloride, N-ethylmaleimide, bafilomycin A1, and chloroquine, increased the intravacuolar pH and promoted the killing of T . whipplei . The ability of T . whipplei to survive in an acidic environment and to interfere with phagosome-lysosome fusion is likely critical for its prolonged persistence in host cells during the course of Whipple's disease . Our results suggest that manipulating the intravacuolar pH may provide a new approach for the treatment of Whipple's disease.

Pol Merkuriusz Lek, 2001 Nov, 11(65), 389 - 93
{Tuberculous pleurisy--still difficult diagnostic problem}; Fijalkowski M et al.; In years 1989-2000 in Lung Diseases Hospital in Lodz tuberculous pleurisy TP was diagnosed in 271 patients; in 130 (48%) cases this diagnosis was confirmed by microbiological and histopathological examinations . Presence of tubercle bacilli in pleural fluid was discovered by smear and culture only in 13 cases, only by culture in 95 and in Bactec method in 7 . During this period in 135 patients with pleural effusion of unknown etiology biopsy of the pleura (with Abrams needle) was performed and specific granuloma was found in 17 (13%) persons . In 15 (12%) patients it was the only method establishing diagnosis of TP (because in 2 remaining patients tubercle bacilli in pleural fluid were also found) . Assessment of clinical symptoms indicates that onset of TP was insidious and course of the disease was subacute in about 30% of patients, manifested mainly by: fatigue, general weakness, night sweats . Radiological estimation of patients with TP disclosed (besides pleural effusion) lung infiltrations in 38 (28%) cases, including 13 (10%) with cavitation . Tuberculin induration with diameter 0 mm was found in 32% of patients with TP and with greater than 10 mm in 29% of patients . Because of low sensitivity of microbiological examinations of pleural fluid for tubercle bacilli biopsy of pleura should be method of choice in the diagnosis of TP in selected cases.

Prikl Biokhim Mikrobiol, 2002 Jan-Feb, 38(1), 103 - 8
{An insoluble colored substrate for dextranase assay}; Khalikova EV et al.; An assay of dextranase (EC 3.2.1.11) was developed by using Sephadex G-200 coupled with Remazol Brilliant Blue (RBB) as an insoluble substrate . The assay procedure included incubation of suspension of the colored substrate in buffer containing enzyme under study, removal of residual insoluble substrate, and measurement of the absorbance of supernatant fluid containing colored soluble hydrolysis products at 595 nm . The procedure was examined in the screening of dextranase-forming bacilli from the microbial collection of the Institute of Biology, Ufa Research Center, RAS.

Clin Microbiol Infect, 1999, 5(10), 617 - 621
Potential source of human exposure to Mycobacterium bovis in Burkina Faso, in the context of the HIV epidemic; Vekemans M et al.; OBJECTIVE: To identify potential sources of human Mycobacterium bovis infection in Bobo-Dioulasso, Burkina Faso . METHODS: A tuberculin survey among 174 cattle was performed . Mycobacteriologic identification in 64 samples of pooled milk, and in 199 tissue samples collected from the slaughterhouse of Bobo-Dioulasso, Burkina Faso, was also done . We retrospectively analyzed the distribution of tuberculosis (TB) cases on 1140 clinical records according to professional occupation and to ethnic group . The frequency of pulmonary and extrapulmonary TB was related to potential exposure and route of transmission of M . bovis from animals . RESULTS: Out of six herds (total 170 bovines), only one was free of any positive tuberculin test . Among 199 bovines which had been slaughtered over four consecutive nights, 38 (19%) had morphologic lesions suggestive of TB; 17 (45%) of those were positive for acid-fast bacilli by microscopic examination on one of their lesions, and 20 samples (53%) presented a positive culture for a pathogenic mycobacterium, including M . bovis and M . tuberculosis . In the retrospective analysis, Peuls more frequently had a pulmonary form of disease . This may be related to the route of transmission . CONCLUSIONS: Attention has to be paid to human TB of bovine origin in Burkina Faso . The identification of M . tuberculosis in milk and in tissue samples raises the question of the transmission of TB from humans to cattle.

Clin Infect Dis, 2002 Mar 15, 34(6), 760 - 6 Epub 2002 Feb 07.
Antibiotic-resistant gram-negative organisms in pediatric chronic-care facilities; Lidsky K et al.; This study was designed to define the prevalence of colonization with antibiotic-resistant gram-negative rectal specimens were obtained from subjects residing in 2 pediatric extended-care facilities and were processed to identify gram-negative organisms resistant to ceftazidime, gentamicin, meropenem, and piperacillin-tazobactam . Horizontal transmission was assessed by analyzing all resistant isolates by pulsed-field gel electrophoresis . Forty percent of subjects were colonized with >/=1 resistant bacillus; >60% of organisms were resistant to >/=2 of the antibiotics tested . Colonization was disproportionate among residents with a tracheostomy or other prosthesis . More than 65% of colonized subjects shared 1 organism with another resident, with cross-colonization occurring among both enteric and nonenteric species . Children residing in chronic-care facilities represent a large reservoir for resistant bacilli . Such colonization may be amenable to simple barrier infection-control procedures.

Lett Appl Microbiol, 2002, 34(2), 86 - 90
A rapid staining technique for the detection of the initiation of germination of bacterial spores; Hamouda T et al.; AIMS: We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination . METHODS AND RESULTS: Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties . Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue . Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination . Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink . CONCLUSIONS: Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment . SIGNIFICANCE AND IMPACT OF THE STUDY: This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.

Respirology, 2001 Dec, 6(4), 317 - 22
Progressive rise of Mycobacterium tuberculosis resistance to rifampicin and streptomycin in Riyadh, Saudi Arabia; Al-Hajjaj MS et al.; OBJECTIVE: The aim of this study was to investigate, for the first time, the factors associated with resistance to antituberculous drugs in Saudi Arabia, and to follow the long-term trends in drug resistance . METHODOLOGY: A retrospective study of patients with positive Mycobacterium tuberculosis recorded at the Riyadh Tuberculosis Center in 1990 was undertaken . The resistance figures from the same centre for the period July 1996 to June 1997 were reviewed for comparison . RESULTS: Resistance was significantly higher in those previously treated (71%) than in those who denied previous treatment (34%) . There was a trend towards association of resistance with cavitatory, multilobar, and acid fast bacilli-positive cases . Nationality (Saudis, Yemenis, others) had no significant effect on resistance . The Riyadh Region now has the same high prevalence of rifampicin resistance as previously reported in the Western Region of the Kingdom . The figures on resistance for the years 1986-88, 1990, and 1996-97 were: isoniazid 19.5/13.8/11.1%, rifampicin 10/20.7/24.6%, streptomycin 5/22/27.4%, ethambutol 3.7/3.9/1.8%, respectively . The reduction in isoniazid and ethambutol resistance coincided with a rise in resistance to rifampicin and streptomycin . We speculate that this resulted from the fact that isoniazid and ethambutol are restricted only to the treatment of tuberculosis and cannot, by law, be dispensed by general practitioners or private pharmacies . Rifampicin and streptomycin, however, are widely used for brucellosis; an endemic disease in Saudi Arabia where up to 12 weeks of rifampicin therapy is recommended . CONCLUSIONS: There has been a significant increase in rifampicin and streptomycin resistance in Saudi Arabia over the last 10 years . Possible causes include poor compliance and wide use of these two drugs for non-tuberculosis conditions . These findings could jeopardize the benefits of the directly observed therapy short course policy which is being implemented in Saudi Arabia . Consideration should be given to prohibiting the routine use of rifampicin for the treatment of brucellosis.

Clin Microbiol Infect, 2001 Dec, 7(12), 709 - 11
Rapid method of extraction and analysis of extended-spectrum beta-lactamases from clinical strains of Klebsiella pneumoniae; Paterson DL et al.; The extraction of periplasmic beta-lactamases from Gram-negative bacilli is a necessary preliminary step to analytical isoelectric focusing . Previously described methods are time-consuming and require large amounts of broth . We describe a lysozyme-based method which needs just 5 mL broth and requires less than 24 h to perform . The method was reproducible in extracting beta-lactamases from reference strains containing known beta-lactamases . We applied the method to a collection of more than 70 extended-spectrum beta-lactamase-producing isolates from a multinational study of bacteremic isolates of Klebsiella pneumoniae . Further studies are being undertaken to assess the method's applicability to other bacterial species.

Acta Cytol, 2002 Jan-Feb, 46(1), 35 - 9
Fine needle aspiration diagnosis in HIV-related lymphadenopathy in Mangalore, India; Shenoy R et al.; OBJECTIVE: To evaluate the usefulness of fine needle aspiration (FNA) study of lymph nodes in HIV-positive patients . STUDY DESIGN: The study was conducted at Kasturba Medical College, Mangalore, India . Samples from lymph nodes of 48 HIV-positive patients were taken and air dried, and wet smears were made . After staining with routine cytologic stains and special stains, detailed cytomorphologic study was conducted . RESULTS: Tuberculosis accounted for nearly half (48%) the cases,followed by HIV lymphadenitis (36%), lymphoma (10%), suppurative lymphadenitis (2%), Mycobacterium avium-intracellulare lymphadenitis (2%) and metastases (2%), in descending order of their frequency . A suppurative picture, which was found in 13% of cases of tuberculous lymphadenitis in AIDS patients, occasionally was misleading without the help of acid fast bacilli stain . CONCLUSION: FNA is a useful tool in the diagnosis of lymphadenopathy in HIV-positive patients provided that proper safety measures are taken to avoid contracting the infection.

Rev Prat, 2001 Dec 1, 51(19), 2069 - 74
{Liver abscess}; Silvain C; Liver abscesses are bacterial or parasitic . In a patient presenting with fever, fatigue and upper right abdominal pain, abdominal ultrasound must be performed to reveal a liver mass . Clinical history and serologies allow presumptive diagnosis . Pyogenic abscesses are mostly secondary to biliary tree infection . Pus aspiration and culture confirm the diagnosis . Gram-negative bacilli and anaerobes are the most frequent germs . Percutaneous drainage combined with antibiotics is the standard treatment . Earlier diagnosis, better techniques for bacterial culture and adapted treatment have improved prognosis . Amoebic abscesses are less frequent but their incidence is increasing in the presence of immunosuppressed state . The diagnosis is based on abdominal ultrasound and serology . Treatment is principally medical.

Pneumologie, 2002 Feb, 56(2), 103 - 7
{Induction of direct antimicrobial activity through mammalian toll-like receptors}; Stenger S et al.; Drosophila, the toll gene controls a powerful innate defense system against bacteria and fungi . Conserved through evolution, the mammalian innate immune system retains a family of homologous Toll-like receptors (TLRs) that are activated by microbial ligands to release cytokines that instruct the adaptive immune responses . Here we show that TLR2 activation leads to killing of intracellular Mycobacterium (M.) tuberculosis in both mouse and human macrophages . In mouse macrophages, bacterial lipoprotein activation of TLR2 leads to a nitric oxide-dependent killing of intracellular tubercle bacilli . In human monocytes and alveolar macrophages, bacterial lipoproteins similarly activated TLR2 to kill intracellular M . tuberculosis, however by an antimicrobial pathway that is nitric oxide independent . TLR2+CD14+CD68+ macrophages were detected in human lesions of tuberculous lymphadenitis within granulomas and surrounding foci of necrosis . These data provide evidence that mammalian TLRs have retained not only the structural features of Drosophila Toll that allow them to respond to microbial ligands, but also the ability directly to activate antimicrobial effector pathways at the site of infection.

Am J Physiol Lung Cell Mol Physiol, 2002 Mar, 282(3), L546 - 55
M . tuberculosis induction of matrix metalloproteinase-9: the role of mannose and receptor-mediated mechanisms; Rivera-Marrero CA et al.; Mycobacterium tuberculosis (Mtb) infection induces the expression of matrix metalloproteinase-9 (MMP-9) in mouse lungs . In cultured human monocytic cells, Mtb bacilli and the cell wall glycolipid lipoarabinomannan (LAM) stimulate high levels of MMP-9 activity . Here, we explore the cellular mechanisms involved in the induction of MMP-9 by Mtb . We show that infection of THP-1 cells with Mtb caused a fivefold increase in MMP-9 mRNA that was associated with increased MMP-9 activity . MMP-9 induction was dependent on microtubule polymerization and protein kinase activation and was associated with increased DNA binding by the transcription factor activator protein-1 (AP-1), which appeared to be important for MMP-9 expression . We then explored the surface molecules potentially involved in Mtb induction of MMP-9, focusing on ligands of the mannose and beta-glucan receptors . MMP-9 activity was induced by the mannose receptor ligands mannan, zymosan, and LAM, whereas the beta-glucan receptor ligand laminarin was not effective . The most active inducers of MMP-9 activity were the particulate ligand zymosan and LAM . Pretreatment of cells with an anti-mannose receptor monoclonal antibody, but not anti-complement receptor 3, decreased the induction of MMP-9 activity by Mtb bacilli . Together, these results suggest that MMP-9 induction by Mtb occurs by receptor-mediated signaling mechanisms involving the binding of mannosylated ligands to mannose receptors, the modulation by cytoskeletal elements such as microtubules, the activation of protein kinases, and transcriptional activation by AP-1.

Vestn Ross Akad Med Nauk, 2001, (11), 11 - 4
{Legionnaires disease: results of 25-years of studying infections, problems and perspectives of research}; Tartakovskii IS; The paper summarizes the results of 25-year study of Legionnaires' disease (Legionnaire's bacillus infection) in its epidemiology, diagnosis, and prevention, the taxonomy and biology of the causative organism . Emphasis is laid on the contribution of the Russian school of investigators in studying this problem . Despite the intensive efforts in investigating the infection and its causative organism, there are still many problems in predicting and preventing the epidemics of Legionnaires' disease, which annually occur in different countries of the world . Legionnaires' disease is a typical example of technogenic infections: active industrial and domestic uses of closed water circulating systems in industry and aerosol sources promote the spread of the infection caused by Legionnaire's bacillus and make its control in these systems . Despite the introduction of current immunological and molecular genetic methods, diagnosis is effective only for L . pneumophila, serogroup 1 . The sensitivity and specificity of methods for diagnosis and identification for other 15 L . pneumophila serogroups and 41 species of L . bacilli are far from being perfect . L . bacilli are also a most popular object for studying the biochemical and genetic aspects of its intracellular parasitism.

Rev Clin Esp, 2001 Dec, 201(12), 696 - 700
{Etiology of hemoptysis: Prospective analysis of 752 cases}; Haro Estarriol M et al.; OBJECTIVES: To know the etiology of hemoptysis and its distribution in our area compared with other current and historic series . METHODS: Prospective, descriptive study of 752 cases of hemoptysis . The etiology of these cases was determined by evaluating the clinical record, chest X-ray, functional respiratory tests and sputum smear for acid-fast bacilli, together with other examinations such as bronchoscopy and computerized tomography (CT) according to previously established criteria . RESULTS: The mean age of patients was 60 years (SD: 15 years), 79% were males and 75% smokers . Diagnoses included neoplasms (28%), chronic bronchitis (19.8%), bronchiectasis (14.5%), pneumonia or lung abscess (11.5%), idiopathic forms (8%), and miscellaneous causes with lower percentages such as tuberculosis (1.7%) and its sequelae (4%), cardiac diseases (1.5%) and pulmonary embolism (2.3%) . Bronchoscopy was performed in 694 (92.2%) and detected the bleeding site or provided a specific diagnosis in 39% (81% if only neoplasms are considered) and non-specific but abnormal in 57% . The amount of bleeding was small in most cases and kept no relationship with causes . CONCLUSIONS: Neoplasms, chronic bronchitis and bronchiectasis are the most common causes of hemoptisis in our area . Bronchiectasis and tuberculosis have decreased compared with previous series used as reference . In contrast, there has been an increase in the number of neoplasms and chronic bronchitis and a plateau in those attributed to cardiac diseases, pneumonia or lung abscess and cryptogenetic forms . To obtain the origin, bronchoscopy complemented by CT remains essential.

Hong Kong Med J, 1998 Mar, 4(1), 63 - 66
A case of colonic tuberculosis mimicking Crohn's disease; Lau CF et al.; A 26-year-old Thai woman who has lived in Hong Kong for the past 3 years presented with a 2-month history of feverishness, intermittent epigastric discomfort, anorexia, and weight loss . She had had per rectal bleeding for 10 days . Colonoscopy on two separate occasions revealed multiple ulcerations involving the entire colon, with rectal sparing . Histological examination of the two sets of colonic biopsies that were obtained during colonoscopy suggested Crohn's disease . There was no response to mesalazine and metronidazole, but the patient responded promptly to a therapeutic trial of antituberculous drugs . Cultures from the first set of colonic biopsies were negative for acid-fast bacilli, but 8 weeks after the second colonoscopy, cultures from the second set of biopsies yielded Mycobacterium tuberculosis . This case illustrates that the diagnosis of colonic tuberculosis requires a high index of suspicion . In cases where the information available does not reveal a definite differentiation between colonic tuberculosis and Crohn's disease, corticosteroids should be withheld . The administration of corticosteroids to a patient with colonic tuberculosis may have disastrous results, and a therapeutic trial of antituberculous drugs should be considered instead.

Zhonghua Wai Ke Za Zhi, 2000 May, 38(5), 369 - 71
{Pathogenic microbes of biliary infection and their resistance to antibiotics}; Zhu J et al.; OBJECTIVE: To understand the change of the main pathogenic microbes of biliary infection and their resistance rates to 12 antibiotics so as to instruct rational application of antibiotics clinically . METHODS: All isolates were identified by analytic products incorporation (API) bioMerieux . The susceptibility of antibiotics to 470 pathogenic microbes was tested by minimal inhibition concentration (MIC) . RESULTS: There were 470 pathogenic microbes with positive biliary culture . The percentage of Escherichia coli, intestinal cocci and Klebsiella pneumoniae was 129 (27.4%), 82 (17.4%) and 76 (16.2%), respectively . In the resistance rates of the gram negative bacilli to 12 antibiotics, the lowest was 4.0% for imipenem followed by . 14.1% Ceftazidime and 16.9% amikacin . In the resistant rates of the gram positive cocci to 12 antibiotics, and had the lowest rate was 6.0% for vancomycin and 16.4% for imipenem . CONCLUSIONS: The main pathogenic microbes of biliary infection were Escherichia coli, intestinal cocci and Klebsiella pneumoniae . Imipenem is the first used for biliary infection in dealing with multiresistant intestinal cocci.

Expert Opin Investig Drugs, 2002 Feb, 11(2), 189 - 215
The macrolides; Blondeau JM et al.; Erythromycin, which was introduced over 50 years ago, was the first macrolide to be used clinically . "New" macrolides, for the treatment of patients with various infectious diseases, were not clinically introduced until 40 years later . The pharmacokinetic and adverse events profile of erythromycin initially limited its use to an alternative agent for patients with allergy to beta-lactam agents . However, the emergence of atypical and/or new pathogens and the ongoing escalation of acquired antimicrobial resistance has impacted on the empirical and organism directed therapy of infectious diseases . Azithromycin and clarithromycin were developed by enhancing the basic macrolide structure . Some of the basic features associated with these new agents include a pharmacokinetic profiles that allow once or twice daily dosing with a much lower incidence of side effects and a substantially broader spectrum of activity which includes some Gram-negative bacilli, atypical pathogens and new, unconventional or uncommon pathogens . Clinical trial data has supported the use of "new" macrolides in a wide range of clinical indications, however, some specific indications are currently restricted to treatment with either azithromycin or clarithromycin . Macrolide resistance is a class effect and depending on the mechanism will confer either low or high level resistance . While resistance is problematic, it does not always result in clinical failure . The macrolides are a valuable class of antimicrobial agent and play an important role in the management of infectious diseases.

J Antibiot (Tokyo), 2001 Nov, 54(11), 926 - 33
Pyloricidins, novel anti-helicobacterpylori antibiotics produced by Bacillus sp . I . Taxonomy, fermentation and biological activity; Nakao M et al.; Novel anti-Helicobacter pylori antibiotics, pyloricidins A, A1, A2, B, C and D, were discovered in the culture broth of two bacilli strains . Pyloricidins selectively inhibited the growth of H . pylori . Pyloricidin B was efficacious in the treatment of gastric infection caused by H . pylori in Mongolian gerbils and may be promising for cure of H . pylori infection as a single agent.

J Clin Microbiol, 2002 Feb, 40(2), 508 - 11
Improving sensitivity of direct microscopy for detection of acid-fast bacilli in sputum: use of chitin in mucus digestion; Farnia P et al.; In order to try to improve the results of direct smear microscopy, we used the mucus-digesting quality of chitin in tuberculosis (TB) laboratories . For this purpose, a total of 430 sputum specimens were processed by the N-acetyl-L-cysteine concentration, sodium hypochlorite (NaOCl) liquefaction, chitin sedimentation, and direct microscopy methods . Then, the smear sensitivity for acid-fast bacillus detection by chitin-treated sputum was compared with the sensitivity of smears prepared by other methods . Our results showed that the chitin solution took less time to completely homogenize the mucoid sputum than did the N-acetyl-L-cysteine and NaOCl methods . The N-acetyl-L-cysteine concentration method demonstrated sensitivity and specificity levels of 83 and 97%, respectively . In comparison, the sensitivity of chitin sedimentation was 80%, with a specificity of 96.7% . The NaOCl liquefaction method showed a sensitivity of 78%, with a specificity of 96% . Finally, the sensitivity of direct microscopy was lower than those of the other tested methods and was only 46%, with a specificity of 90% . The chitin and NaOCl liquefaction methods are both easy to perform, and they do not require additional equipment (centrifuges) . Also, our results demonstrated that the chitin method is less time-consuming than the NaOCl method, since only 30 min of incubation is required to bring complete sedimentation of bacilli in chitin-treated sputum whereas the NaOCl method needs 10 to 12 h to give the same results in the same sputum specimens . Therefore, the chitin liquefaction and sedimentation method may provide better results in TB laboratories of developing countries than the N-acetyl-L-cysteine concentration, NaOCl overnight sedimentation, and direct smear microscopy methods.

J Clin Microbiol, 2002 Feb, 40(2), 501 - 7
Rapid detection of pyrazinamide-resistant Mycobacterium tuberculosis by a PCR-based in vitro system; Suzuki Y et al.; Pyrazinamide (PZA), an analog of nicotinamide, is a prodrug for tuberculosis which requires conversion to the bactericidal compound pyrazinoic acid by bacterial pyrazinamidase activity . Mutations leading to a loss of pyrazinamidase activity cause PZA resistance in Mycobacterium tuberculosis . Thus, the detection of pyrazinamidase activity makes the discrimination of PZA-resistant tuberculosis possible . However, the detection of the pyrazinamidase activity of M . tuberculosis isolates needs a large amount of bacilli and is therefore time consuming . In this paper, we describe a new method for the detection of pyrazinamidase activity with a PCR-based system . The genes encoding pyrazinamidase (pncA genes) in 30 resistant clinical isolates were amplified by PCR by using forward primers containing bacteriophage T7 promoter sequences at their 5" ends . Then the PCR products were directly subjected to an in vitro transcription-translation coupled system . All of the PZA-resistant isolates tested showed reduced pyrazinamidase activity compared to susceptible M . tuberculosis type strain H37Rv . In contrast, all of the 15 susceptible clinical isolates exhibited pyrazinamidase activities similar to that of H37Rv . This fact suggested the possibility of the usefulness of this system for the rapid detection of PZA-resistant M . tuberculosis.

Am J Clin Dermatol, 2002, 3(1), 37 - 46
The role of fluoroquinolones in skin and skin structure infections; Blondeau JM; Skin and skin structure infections encompass a broad range of clinical presentations and disease severity . Antimicrobial therapy is clearly beneficial for both recovery from these infections as well as preventing disease progression . Fluoroquinolones are potent broad spectrum antimicrobial agents with the older agents characterized as having broad spectrum anti-Gram-negative activity, borderline activity against clinically important Gram-positive pathogens and little or no anti-anaerobic bacteria activity . In contrast, the new quinolones are characterized by having enhanced activity against Gram-positive pathogens, anti-anaerobic activity and they remain highly active against aerobic-Gram-negative bacilli . Several fluoroquinolones have been evaluated for the treatment of uncomplicated skin and soft tissue infection, difficult skin and soft tissue infection and serious skin and skin structure infections . Clinical cure rates were found to be equivalent to comparators suggesting a role for the fluoroquinolone in treating these infections . It may be necessary to use some fluoroquinolones in combination with anti-anaerobic agents for those infections with mixed aerobic and anaerobic pathogens . Some additional clinical trials are necessary to identify the full potential of newer fluoroquinolones for skin and skin structure infections . At present, quinolones are, in general, equivalent to beta-lactam agents in the treatment of skin and soft tissue infection.

Curr Opin Investig Drugs, 2001 Feb, 2(2), 198 - 201
Preclinical candidates and targets for tuberculosis therapy; Barry CE 3rd; Like many neglected diseases of the developing world, tuberculosis (TB) has a thin portfolio of new compounds currently in the discovery pipeline with near-term clinical potential . Co-development of broad-spectrum antibacterials for TB indications is superficially attractive but unlikely to result in significant advances in therapy . Genomic information has been useful in the redesign of second-line antituberculars such as ethambutol and such molecules will likely soon enter preclinical development . New targets and lead compounds with activity against the mycobacterial cell wall and non-replicating bacilli are the subject of current discovery programs.

Hepatogastroenterology, 2001 Nov-Dec, 48(42), 1716 - 20
Monocyte CD14 changes and endotoxemia after major hepatectomy; Komatsu S et al.; BACKGROUND/AIMS: This study was designed to ascertain whether decreases in membrane CD14 (mCD14), the endotoxin receptor on the surface of the monocyte cell membrane, reflect the presence of endotoxemia, and whether endogenous endotoxemia is present after hepatectomy . METHODOLOGY: First, in cases with a definitive clinical diagnosis of endotoxemia, we determined the serum endotoxin level by the ES (Endospesy) method, from the percentage of cells positive for membrane CD14, from the CD14 fluorescence intensity (MFI-CD14), and from the quantification of soluble CD14 (sCD14); and calculated the diagnostic accuracy obtained with each . Next, in 13 cases of extended hepatectomy (in excess of lobectomy) the mCD14-positive rates were determined around the time of the operation, and compared with the clinical courses of infection foci, if any, and other findings . RESULTS: The diagnostic accuracy of endotoxemia obtained by ES, mCD14, MFI-CD14 and sCD14 were 77.3%, 95.5%, 86.7% and 66.7%, respectively, so that the highest was given by the positive mCD14 cell rate . Although no infectious complication was detected in any of the 9 patients in whom the mCD14-positive rates changed within the normal range, of the 4 patients in whom the mCD14-positive rates fell, 2 had cholangitis due to gram-negative bacilli, one who was infected by central vein catheterization, was thought to have exogenous endotoxemia; and the fourth, in whom no clear focus of infection was detected, ran a temperature of 38 degrees C (over 100 degrees F) of unknown origin for 16 days . In this last patient, gram-negative bacilli were detected in both the saliva and the gastric juice, and so endogenous endotoxemia was suspected . CONCLUSIONS: These results show that decrease in the mCD14-positive rate is more accurately diagnostic of endotoxemia than the endotoxin value obtained by the ES method, and was considered to be a definitive diagnosis of endotoxemia . In addition, from the reduction of the mCD14-positive rate after extended hepatectomy, it was considered that endogenous endotoxemia occurred in one case out of 13 (8%).

J Infect Chemother, 2001 Sep, 7(3), 186 - 90
Mycobacterium haemophilum infection in a Japanese patient with AIDS; Endo T et al.; Mycobacterium haemophilum has been described as a pathogen that causes cutaneous lesions in immunocompromised patients . A specimen from a skin ulcer on the leg of a Japanese patient with acquired immunodeficiency syndrome yielded acid-fast bacilli on blood agar plates after 4 weeks of incubation at 30 degrees C, but the organism was not found on Ogawa egg slants . The organism was identified as M . haemophilum, on the basis of 16S rRNA gene sequence analysis . Prolonged culture in an optimal environment that includes an iron supplement, and growth temperatures at 28 degrees to 33 degrees C are necessary to grow M . haemophilum . Genotypic characterization of 16S rRNA is useful for a rapid diagnosis of this slowly growing mycobacterium.

Skeletal Radiol, 2002 Jan, 31(1), 1 - 8 Epub 2001 Nov 09.
Multifocal musculoskeletal tuberculosis in children: appearances on computed tomography; Morris BS et al.; PURPOSE AND PATIENTS: The incidence of skeletal tuberculosis (TB), which once accounted for a majority of cases of extrapulmonary tuberculosis, has fallen significantly in recent years with the advent of effective drug therapy . Disseminated bone involvement in TB is very uncommon but it may still occur in countries where TB is endemic . We present the imaging findings of four children ranging in age from 2 to13 years, each of whom had multiple osseous stigmata of tuberculous infection . They presented to us over a period of 9 months . RESULTS AND CONCLUSION: Three of four children had calvarial lesions, with involvement of the bony orbit in one, and large abscesses were present in the chest wall and the mediastinum of another . Lesions along the dorsal spine were demonstrated in three cases, two of which showed epidural extensions . Bone lesions in the thoracic cage accompanying those in the spine were also seen in two children, one of whom had a solitary destructive focus in a rib distant from the site of vertebral involvement . Bone lesions involving the first metacarpal in one case and the scapular wing in two others are also described . The diagnosis in each of the cases was confirmed by the identification of epitheloid giant cells and caseous necrosis or tubercle bacilli in fine needle aspirates or on tissue culture studies.

Kekkaku, 2001 Dec, 76(12), 723 - 8
{Treatment outcomes of multidrug-resistant tuberculosis--comparison between success and failure cases}; Shigetoh E et al.; The thirty-two times of treatment in 27 patients with multidrug-resistant tuberculosis (MDR-TB) were analyzed retrospectively . In twenty-eight times of treatments cases had previous histories of antituberculosis chemotherapy . Drug sensitivity tests were performed by Microtiter method for isoniazid (INH), rifampicin (RFP), ethambutol, streptomycin, kanamycin, enviomycin, ethionamide, para-aminosalicylic acid and cycloserine . A drug is defined as 'active drug' when the drug was proved to be sensitive by the drug sensitivity tests or never used in the past or used for not more than 2 months in case of pyrazinamide (PZA) and less than one month for fluoroquinolones . Outcomes of treatments were grouped as follows; A: bacteriologically negative for more than 24 months, B: bacteriologically negative for more than 6 months but less than 24 months, C: bacteriologically relapsed after negative conversion, D: continuously bacilli positive for M . tuberculosis . Mean age of patients in each group were; 61.0 yrs for group A (n = 10), 61.0 yrs for group B (n = 7), 52.5 yrs for group C (n = 4), 57.9 yrs for group D (n = 11) . All patients had cavitary disease and positive sputum smears for acid-fast bacilli . Mean numbers of 'active drugs' used per treatment in each group, were 3.6, 3.3, 2.5 and 1.8 respectively, while the mean number of resistant drug including INH and RFP were 2.8, 3.3, 2.5 and 3.7 . The number of drugs, which was unable to use due to toxicity, were 0.20, 0.14, 0.50, and 0.73 per treatment respectively . All of 9 patients treated with four 'active drugs' were in group A or B and succeeded to achieve negative conversion . The duration of chemotherapy in group A was 13 to 44 months . Treatment had failed in 4 out of 11 patients treated with 3 'active drugs' and 11 out of 12 patients treated with less than 2 'active drugs' . Fluoroquinolones (ofloxacin, levofloxacin or sparfloxacin) were used in 7 out of 10 patients in group A and in 6 out of 9 patients treated with four-drug regimens while they were used only in 3 out of 11 patients in group D . Regimens with at least 4 sensitive drugs are mandatory for the successful treatment of MDR-TB and fluoroquinolones are needed in the majority of cases to ensure the four-drug regimen, because of frequent drug resistance or toxicity to other antituberculosis drugs.

Kekkaku, 2001 Dec, 76(12), 717 - 21
{Clinical analysis of multidrug-resistant tuberculosis}; Yagi T et al.; Forty-three patients with multidrug-resistant tuberculosis at National Chiba-Higashi Hospital were studied retrospectively . TB cases excreting tubercle bacilli which are resistant to both 0.1 microgram/ml of isoniazid and 50 micrograms/ml of rifampicin were defined as multidrug-resistant cases . From 1993 to 1997, we experienced 1627 patients with pulmonary tuberculosis, and among them 43 patients (23-79 years old, 35 males and 8 females) were proved to be multidrug-resistant . Six cases were initially treated cases and other 37 cases had been treated previously . On admission, 40 out of 43 cases (93.0%) were smear positive by sputum examination of mycobacteria and 38 out of 43 cases (88.4%) had cavitary lesions on chest X-ray . Six patients were complicated with diabetes mellitus, two with cancer, one with alcohol dependence, one with chronic hepatitis, and others did not have prominent complications . Three operated patients were cured, the fact shows that the surgical treatment is still a useful measure for cases with the indication . Sixteen patients were cured, eight were still under treatment, and thirteen were died of tuberculosis . One of reasons of poor prognosis of multidrug-resistant tuberculosis is that multidrug-resistant tubercle bacilli are usually resistant to other drugs, too . In case of multidrug-resistant tuberculosis, patients were obliged to be treated in a hospital long-term to prevent the spread of tubercle bacilli . Therefore, it is very important to find out new tuberculosis cases as an early as possible, treat them with proper regimen and prevent dropout by directly observed therapy, thus preventing the emergence of multidrug-resistant tuberculosis . Development of new antituberculous agents is strongly expected.

Ann N Y Acad Sci, 2001 Dec, 953, 151 - 6
The genetics of host resistance and susceptibility to tuberculosis; Davies P et al.; The study of human genomics has the potential to aid our understanding of the interindividual and interpopulation differences in susceptibility to tuberculosis . Resistance to infection is affected by the ability of macrophages to phagocytose and destroy the bacilli . Several genes are involved in this process, and two have been the focus of recent interest: the natural resistance-associated protein (NRAMP1) gene and the genes coding for the vitamin D receptor . Susceptibility genes have also been discovered--for example, one on the X chromosome that may explain the increased susceptibility of males to tuberculosis . Studies have also focused on the variations in virulence of the bacillus in both its drug-susceptible and drug-resistant forms . These mechanisms must be understood in order to prevent, or combat, the emergence of a virulent, multidrug-resistant form of the bacillus that would be uncontrollable by means of today's treatment strategies.

Cornea, 2002 Jan, 21(1), 123 - 5
Detection of mycobacterium tuberculosis by nested polymerase chain reaction in a case of subconjunctival tuberculosis; Biswas J et al.; PURPOSE: To highlight the importance of nested polymerase chain reaction (PCR) in the detection of Mycobacterium tuberculosis in a case of subconjunctival tuberculosis . METHODS: We report a case of a 60-year-old man with subconjunctival nodule in the right eye for duration of 6 weeks . Biopsy of the nodule showed a granuloma with extensive caseation necrosis . Ziehl Neelsen staining for acid-fast bacilli (AFB) was negative . However, because of a strong suspicion of Mycobacterium infection, PCR for M . tuberculosis genome was done, using the nested PCR technique . RESULTS: Polymerase chain reaction for M . tuberculosis showed amplification of Mycobacterium tuberculosis genome with the nested PCR technique . CONCLUSION: Our case indicates that PCR can be a valuable tool in the diagnosis of conjunctival tuberculosis from paraffin sections.

J Immunol, 2002 Feb 1, 168(3), 1309 - 14
Surfactant protein D inhibition of human macrophage uptake of Mycobacterium tuberculosis is independent of bacterial agglutination; Ferguson JS et al.; The innate immune system in the lung is essential for controlling infections due to inhaled pathogens . Mycobacterium tuberculosis (M.tb) encounters components of the innate immune system when inhaled into the lung, but the consequences of these interactions are poorly understood . Surfactant protein D (SP-D) binds to and agglutinates M.tb bacilli, and reduces the uptake of the bacteria by human macrophages . In the current studies, we utilized a recombinant SP-D variant (CDM) that lacks the collagen domain to further characterize the interaction of SP-D with M.tb, and determine the effects of agglutination on bacterial uptake by human monocyte-derived macrophages . These studies demonstrate that the binding of SP-D and CDM to M.tb is saturable and inhibited by carbohydrate competition and Ca(2+) chelation, implicating the carbohydrate recognition domain in the interaction . Fluorescence microscopy reveals that dodecameric SP-D leads to agglutination of the bacilli, whereas the trimeric CDM does not, demonstrating that the multivalent nature of SP-D is essential for agglutination of M.tb . However, preincubation of M.tb with increasing concentrations of SP-D or CDM leads to a concentration-dependent reduction in the uptake of the bacteria by macrophages, indicating that agglutination does not play a direct role in this observation . Finally, the reduced uptake of M.tb by SP-D is associated with reduced growth of M.tb in monocyte-derived macrophages . These studies provide direct evidence that the inhibition of phagocytosis of M.tb effected by SP-D occurs independently of the aggregation process.

J Immunol, 2002 Feb 1, 168(3), 1294 - 301
Dendritic cells are host cells for mycobacteria in vivo that trigger innate and acquired immunity; Jiao X et al.; In the present study, we investigated in vivo the infection and APC functions of dendritic cells (DC) and macrophages (Mphi) after administration of live mycobacteria to mice . Experiments were conducted with Mycobacterium bovis bacillus Calmette-Guerin (BCG) or a rBCG expressing a reporter Ag . Following infection of mice, DC and Mphi were purified and the presence of immunogenic peptide/MHC class II complexes was detected ex vivo on sorted cells, as was the secretion of IL-12 p40 . We show in this study that DC is a host cell for mycobacteria, and we provide an in vivo detailed picture of the role of Mphi and DC in the mobilization of immunity during the early stages of a bacterial infection . Strikingly, BCG bacilli survive but remain stable in number in the DC leukocyte subset during the first 2 wk of infection . As Ag presentation by DC is rapidly lost, this suggests that DC may represent a hidden reservoir for mycobacteria.

Tuberculosis (Edinb), 2001, 81(5-6), 353 - 8
Evaluation of DPPD, a single recombinant Mycobacterium tuberculosis protein as an alternative antigen for the Mantoux test; Campos-Neto A et al.; Although the tuberculin test has aided in the diagnosis of tuberculosis for more than 85 years, its interpretation is difficult particularly because sensitization with non-tuberculous mycobacteria leads to false positive tests . Using the guinea pig model of tuberculosis, we have recently described a recombinant antigen (DPPD) that could circumvent this problem . The DPPD gene is unique to the M . tuberculosis complex organisms and is absent in the organisms representative of all other members of the Mycobacterium genus . Moreover, DPPD induced strong DTH in 100% of the guinea pigs infected with M . tuberculosis and in none of the guinea pigs immunized with nine different species of Mycobacterium . Here we present results of a clinical investigation using DPPD . Mantoux test using both PPD and DPPD was initially performed in 26 patients with confirmed pulmonary tuberculosis and in 25 healthy PPD negative individuals . The results indicated that both PPD and DPPD elicited DTH in 24 out of the 26 patients . No DTH was observed in any of the PPD negative individuals . In addition, a small clinical trial was performed in a population of 270 clinically healthy and randomly selected individuals . DPPD produced a bimodal histogram of skin reaction size and PPD produced a skewed histogram . Because the DPPD gene is not present in non-tuberculous bacilli, these results suggest that this molecule can be an additional tool for a more specific diagnosis of tuberculosis .

J Med Microbiol, 2002 Jan, 51(1), 42 - 9
Conditions that may affect the results of susceptibility testing of Mycobacterium tuberculosis to pyrazinamide; Zhang Y et al.; Pyrazinamide (PZA) is an important front-line anti-tuberculosis drug that is active only at acid pH . However, acid pH causes significant difficulty for PZA susceptibility testing . A common problem in PZA testing is false resistance caused by large bacterial inocula . This study investigated the relationship of false resistance to numbers of bacilli, pH and other factors that potentially affect susceptibility to PZA . Large inocula (10(7-8) bacilli/ml) of M . tuberculosis H37Ra caused significant increase in medium pH from 5.5 towards neutrality, and thus produced false resistance results . The increase in medium pH was determined to be a function of live bacilli; heat-killed bacilli had little or no effect . Susceptibility to PZA and its active derivative pyrazinoic acid (POA) was comparable on 7H11 agar medium, but POA was less active than PZA in liquid medium containing bovine serum albumin (BSA), suggesting that susceptibility to PZA or POA was reduced in the presence of BSA, because of its neutralising effect on medium pH and significant POA binding . A 3-month-old H37Ra culture was shown to be more susceptible to PZA exposure than a 4-day log-phase culture, suggesting that PZA is more active for non-growing bacilli . Finally, reserpine, an inhibitor of POA efflux pump, increased susceptibility to PZA even near neutral pH 6.8, with an MIC of 400 mg/L compared with 1,000 mg/L without reserpine . These findings should have implications for understanding the mode of action of PZA and for PZA susceptibility testing.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 2000 Jul, 20(7), 524 - 6
{Changes of glutamate and gamma-aminobutyric acid contents in brain tissue of brain edema and effects of baicalin on them in rats}; Yin F et al.; OBJECTIVE: To observe the changes of glutamates (Glu) and gamma-aminobutyric acid (GABA) contents in brain tissue of infectious brain edema, and effects of baicalin on them in rats . METHODS: The high performance liquid chromatography (HPLC) was used to determine the Glu and GABA contents in homogenized brain tissue after baicalin was injected intraperitoneally on infectious brain edema induced by pertussis bacilli in rats . RESULTS: Glu contents of homogenized brain tissue in blank control group (C), normal saline group (NS), pertussis bacilli group (PB) and baicalin group (BC) had no significant difference (P > 0.05) . The concentration of GABA was more significantly increased in the BC group (4.06 +/- 0.52) than that in the PB group (2.71 +/- 0.29) (P < 0.05) . Water content of brain was reduced in the BC group (79.8 +/- 0.4) as compared with the PB group (82.2 +/- 0.3)(P < 0.01) . CONCLUSION: Baicalin has a protective effect against the pertussis bacilli-induced brain edema in rats, and protective effect was correlated to the increased content of GABA in the brain tissue in rats.

AIDS Patient Care STDS, 2001 Dec, 15(12), 607 - 10
Chlamydia trachomatis and Mycobacterium tuberculosis lung infection in an HIV-positive homosexual man; Monno R et al.; A 31-year-old homosexual man, who was human immunodeficiency virus (HIV)-positive was admitted for fever and cough . Chest computed tomography (CT) revealed the presence of diffuse interstitial reticular nodulation, and brain nuclear magnetic resonance imaging showed the presence of nodular frontal lesions . Microscopic examination of sputum and other body fluids showed the presence of acid-fast bacilli and culture-only growth Mycobacterium tuberculosis . Serology for respiratory tract pathogens was negative except for Chlamydia . An antibody titer in the immunoglobulin G (IgG) class of 1:64 for Chlamydia pneumoniae and, unexpectedly, an antibody titer of 1:1024 for C . trachomatis were found . The patient was successfully treated with antituberculosis agents, and clarithromycin, for presumptive chlamydial infection.

Hepatology, 2002 Jan, 35(1), 140 - 8
Bacterial infections in cirrhosis: epidemiological changes with invasive procedures and norfloxacin prophylaxis; Fernandez J et al.; The extensive use of invasive procedures and of long-term norfloxacin prophylaxis in the management of cirrhotic patients may have influenced the epidemiology of bacterial infections in cirrhosis . We conducted a prospective evaluation of all bacterial infections diagnosed in patients with cirrhosis in a Liver Unit between April 1998 and April 2000 . A total of 405 patients presented 572 bacterial infections in 507 admissions . Spontaneous bacterial peritonitis was the most frequent infection (138 cases) . Gram-positive cocci were responsible for 53% of total bacterial infections in the study, being the main bacteria isolated in nosocomial infections (59%) . Patients requiring treatment in an intensive care unit and those submitted to invasive procedures presented a higher rate of infections caused by gram-positive cocci (77% vs . 48%, P <.001 and 58% vs . 40%, P <.02, respectively) . Fifty percent of culture-positive spontaneous bacterial peritonitis in patients on long-term norfloxacin administration (n = 93) and 16% in patients not receiving this therapy (n = 414) were caused by quinolone-resistant gram-negative bacilli, P =.01 . The rate of culture-positive spontaneous bacterial peritonitis caused by trimethoprim-sulfamethoxazole-resistant gram-negative bacilli was also very high in patients on long-term norfloxacin administration (44% vs . 18%, P =.09) . In conclusion, infections caused by gram-positive cocci have markedly increased in cirrhosis . This phenomenon may be related to the current high degree of instrumentation of cirrhotic patients . Quinolone-resistant spontaneous bacterial peritonitis constitutes an emergent problem in patients on long-term norfloxacin prophylaxis, with trimethoprim-sulfamethoxazole not being a valid alternative.

Mikrobiologiia, 2001 Nov-Dec, 70(6), 825 - 37
{Amphibacillus fermentum sp . nov., Amphibacillus tropicus sp . nov.--new alkaliphilic, facultatively anaerobic, saccharolytic Bacilli from Lake Magadi}; Zhilina TN et al.; New alkaliphilic, saccharolytic, rod-shaped, gram-positive bacteria resistant to heating and drying and phylogenetically affiliated to the Bacillus lineage were isolated under strictly anaerobic conditions from sediments of the alkaline and highly mineralized Lake Magadi . Strain Z-7792 forms endospores; in strain Z-7984, endospore formation was not revealed . The strains are capable of both anaerobic growth (at the expense of fermentation of glucose and certain mono- and disaccharides with the formation of formate, ethanol, and acetate) and aerobic growth . Among polysaccharides, the strains hydrolyze starch, glycogen, and xylan . Yeast extract or methionine are required for growth . The strains are strict alkaliphiles exhibiting obligate requirement for Na+ and carbonate ions but not for Cl- ion . Growth occurs at a total mineralization as high as 3.3-3.6 M Na+, with an optimum at 1-1.7 M Na+ . Strain Z-7792 is an obligate alkaliphile with a pH growth range of 8.5-11.5 and an optimum of 9.5-9.7 . Strain Z-7984 grows in a pH range of 7.0-10.5 with an optimum at 8.0-9.5 . Both strains are mesophiles having a growth optimum at 37-38 degrees C . They belong to bacilli with a low G + C content . The G + C contents of the DNA of strains Z-7792 and Z-7984 are 39.2 and 41.5 mol%, respectively . These isolates of facultatively anaerobic, strictly alkaliphilic, Na(+)-dependent bacilli can be considered representatives of the ecological group adapted to the life at drying-up shoars of soda lakes . Because of their independence of NaCl and lack of obligate dependence on sodium carbonates, the isolates are to be assigned to athalassophilic organisms . According to their physiological and phylogenetic characteristics, they taxonomically belong to group 1 of the species of bacilli, occupying a position intermediate between the genera Amphibacillus and Gracilibacillus . The isolates are described as new species of Amphibacillus: A . fermentum (type strain, Z-7984T) and A . tropicus (type strain, Z-7792T).

Mem Inst Oswaldo Cruz, 2001 Nov, 96(8), 1137 - 9
Inhibition of the polymerase chain reaction by sputum samples from tuberculosis patients after processing using a silica-guanidiniumthiocyanate DNA isolation procedure; Suffys P et al.; With the objective to evaluate PCR-mediated detection of Mycobacterium tuberculosis DNA as a diagnostic procedure for diagnosis of tuberculosis in individuals attending ambulatory services in Primary Health Units of the City Tuberculosis Program in Rio de Janeiro, Brazil, their sputum samples were collected and treated with a DNA extraction procedure using silica-guanidiniumthiocyanate . This procedure has been described to be highly efficient for extraction of different kind of nucleic acids from bacteria and clinical samples . Upon comparing PCR results with the number of acid-fast bacilli, no direct relation was observed between the number of bacilli present in the sample and PCR positivity . Part of the processed samples was therefore spiked with pure DNA of M . tuberculosis and inhibition of the PCR reaction was verified in 22 out of 36 (61%) of the samples, demonstrating that the extraction procedure as originally described should not be used for PCR analysis of sputum samples.

Mem Inst Oswaldo Cruz, 2001 Nov, 96(8), 1129 - 33
Detection of Mycobacterium leprae DNA by polymerase chain reaction in the blood of individuals, eight years after completion of anti-leprosy therapy; Santos AR et al.; Thirty eight patients with indeterminate leprosy (HI), at least 4 to 6 years after discharge from multibacillary (MB) or paucibacillary (PB) schemes of anti leprosy multidrug therapy (MDT), were submitted to traditional diagnostic procedures for leprosy and to polymerase chain reaction (PCR) analysis of different clinical samples for detection of Mycobacterium leprae DNA . No significant difference was observed for any of the parameters analyzed between PB or MB schemes of treatment and no indications were found for more efficient outcome of HI using the MB scheme . Remarkably, 18 (54.5%) of the individuals were PCR positive in at least one of the samples: positivity of PCR was highest in blood samples and four individuals were PCR positive in blood and some other sample . Upon comparison of PCR results with clinical and histopathological parameters, no correlation was found between PCR-positivity and eventual relapse . This is the first report on detection of M . leprae DNA in PB patients, more than half a decade after completion of MDT, suggesting that live bacilli are present and circulating much longer than expected, although reinfection of the individuals can not be excluded . Overall, we feel that because of the high sensitivity of the assay, extreme care should be taken about association of PCR results, efficacy of treatment and disease status.

Zhongguo Zhong Xi Yi Jie He Za Zhi, 1999 Apr, 19(4), 224 - 6
{Effect of baicalin and tetramethylpyrazine on intracranial hypertension of infectious brain edema in rabbits}; Chen G et al.; OBJECTIVE: To evaluate the effects of mannitol (MN), baicalin (BC) and tetramethylpyrazine (TMPZ) on intracranial hypertension (ICH) of infectious brain edema induced by Pertussis bacilli (PB) in rabbits . METHODS: Forty-five rabbits were divided randomly into five groups: the normal saline group (A); the pertussis bacilli group (B); the MN treated group (C); the TMPZ treated group (D) and the BC treated group (E) . PB was injected into the left internal carotid artery of rabbits to establish the infectious brain edema model, their intracranial pressure (ICP) was monitored continuously by fiberoptic ICP monitor . Treatments were given intravenously 1 hour after injecting PB, and changes of ICP, brain water content (WC), Evans blue (EB), Na+, K+ and morphology were observed within 4 hours after PB or NS injection . RESULTS: MN, TMPZ and BC all could reduce the brain edema and ICH in rabbits induced by PB, the effect of MN was the most rapid and obvious one, but it lasted for rather shorter time, while the effect of TMPZ and BC was initiated later but lasted for a longer time . The ICP of Group C in 2nd to 4th hour was higher than that of Group D and Group E, but lower than that of Group B . The ICP of Group E in 3rd to 4th hour was lower than that of group D . At the same time of lowering ICP, brain WC, EB, Na+ contents and pathologic morphologic change were also alleviated . CONCLUSION: Both BC and TMPZ could reduce ICP in infectious brain edema, the effect of BC was better than that of TMPZ.

Zhonghua Jie He He Hu Xi Za Zhi, 2000 Sep, 23(9), 559 - 62
{Study on pathogenicity of sputum from cavity of sputum negative patients with pulmonary tuberculosis after short course chemotherapy}; Feng H et al.; OBJECTIVE: To find out whether or not the sputa of pulmonary tuberculosis(PTB) patients with cavitation but sputum negative have pathogenicity after short course chemotherapy . METHODS: Guinea pigs were inoculated with sputa of PTB patients who were with cavitation but sputum negative after having finished short course chemotherapy . Then their body weight, enlargement of local lymph nodes and other ordinary symptom were observed . Six weeks later, pathological changes of TB in the internal organs were examined by dissecting these guinea pigs . Culture and drug resistance test of tubercle bacillus were also conducted . All of which were with negative and positive controls . RESULTS: Of the 63 cases included, 3(5%) patients' sputa resulted in tuberculous nodulation varying in amount in lung, liver and spleen of these guinea pigs, and the culture for tubercle bacillus of these sputa was positive too . CONCLUSIONS: 5% of sputum collected from PTB patients with cavitation but sputum negative still show pathogenicity after short course chemotherapy . For the cases with drug-resistant PTB and slow sputum negative conversion, the treatment should be prolonged and tubercle bacilli in their sputa should be monitored.

Zhonghua Jie He He Hu Xi Za Zhi, 2000 Sep, 23(9), 556 - 8
{Detecting mycobacteria and their L-forms in peripheral blood from pulmonary tuberculosis patients by cultivation with hemolyzed-centrifugated blood in liquid medium}; Zhu M et al.; OBJECTIVE: To establish a new method of detecting mycobacteria and their L-forms in blood . METHODS: 65 samples of peripheral blood from patients with pulmonary tuberculosis were cultivated directly or cultivated with hemolyzed-centrifugated blood sediments in 92-3TB and 92-3TBL liquid medium, and the cultures were staining with immunoenzyme technique (ABC method) . RESULTS: In 65 specimens, the positive rates of mycobacteria and their L-forms were 15%, 26% respectively with total isolation rate of 32% . In cultivating with hemolyzed-centrifugated blood the positive rate was much higher than direct cultural method (P < 0.05) . In sputum the positive rates of acid-fast bacilli and their L-forms were 38% and 20% respectively, with total isolation rate of 52%, higher than that in peripheral blood (P < 0.05), and the total positive rate of combinative detection of blood and sputum salmples was 65% . CONCLUSIONS: Mycobacteria and their L-forms exist in peripheral blood of patients with pulmonary tuberculosis, and mainly in L-forms . Cultivation with hemolyzed-centrifugated blood in liquid medium and staining with immunoenzyme technique are valuable in routine detection of mycobacteria and their L-forms in peripheral blood . The positive rate of bacterial culture of pulmonary tuberculosis could be increased in combination with sputum mycobacteria and their L-forms examination.

Bull Soc Pathol Exot, 2000 Jan, 93(5), 337 - 9
{Epidemiological aspects of tuberculosis in middle west of Madagascar}; Rakotondramarina D et al.; We conducted a 5-year (1989-1993) retrospective analysis on a series of patients screened and treated for tuberculosis in order to determine the epidemiological aspects of the disease in mid-western Madagascar . Pulmonary forms affected 97% of patients, and predominantly men (sex ratio: 1.4); 83% of patients with pulmonary tuberculosis were coughing up acido-alcoholo-resitant bacilli . Haemoptysis and general deterioration, the most frequently met pathological signs, were observed respectively in 62% and 24% of cases . This explains the high rate of hospitalisation (42%), especially for patients with difficult access to services . Significantly, in terms of socio-professional category, live-stock breeders and farmers represented 32% of notified cases, and cattle-traders 18% . We draw attention to the possible role played by Mycobacterium bovis in human case-findings in a region characterised by cattle-breeding.

Head Neck, 2002 Jan, 24(1), 6 - 15
A pilot study evaluating the safety and microbiologic efficacy of an economically viable antimicrobial lozenge in patients with head and neck cancer receiving radiation therapy; El-Sayed S et al.; BACKGROUND: Mucositis occurs in almost all radiotherapy-treated head and neck cancer patients, in approximately 75% of patients receiving hematopoietic marrow transplantation, and in approximately 40% of all patients who receive chemotherapy . Mucositis is painful, may affect all oral functions, and is a dose- and rate-limiting toxicity of therapy for cancer . Radiation-associated mucositis (onset, intensity, and duration) has been shown in recent clinical trials to be modified by the use of antibacterial/antifungal lozenges . PURPOSE: The aim of this collaborative two-center phase II study was to assess the toxicity and microbiologic efficacy of an economically viable antimicrobial lozenge in the management of patients receiving radiation therapy for head and neck cancer . MATERIALS AND METHODS: Seventeen patients scheduled to receive radical or postoperative radiotherapy were provided with bacitracin, clotrimazole, and gentamicin (BCoG) lozenges (one lozenge dissolved in the mouth qid from day 1 of radiotherapy until completion) . Ease of use and palatability of the lozenges, patients' symptoms (swallowing and pain), and quantitative and qualitative microbiologic evaluation of an oral rinse collection was conducted at least once weekly during radiation therapy . RESULTS: No significant side effects were reported from the use of the lozenges . The lozenges were well tolerated at the beginning of treatment by all patients, with some minor difficulty associated with oral discomfort toward the end of the treatment . Microbiologic evaluation showed consistent elimination of yeast organisms in all patients . In four patients there was no growth of gram-negative bacilli on culture, whereas in two patients, fluctuating counts were seen, and one patient had increased counts . The remaining patients had significant reduction in the gram-negative bacilli counts . CONCLUSIONS: This study demonstrated that the BCoG lozenge is tolerable and microbiologically efficacious, achieving elimination of Candida in all patients and reduction in gram-negative flora in most patients . A phase III study is underway to evaluate the clinical efficacy of this lozenge .

Int J Tissue React, 2001, 23(4), 113 - 25
Virulence of Mycobacterium tuberculosis affects interleukin-8, monocyte chemoattractant protein-1 and interleukin-10 production by human mononuclear phagocytes; Fietta A et al.; Microbial virulence and cytokine-mediated immune responses to Mycobacterium tuberculosis infection are important determinants of the pathogenesis of human tuberculosis . To determine the interrelationship between mycobacterial virulence and cytokine induction, human monocytes and monocyte-derived macrophages were infected with attenuated (H37Ra) and virulent (H37Rv and CH306) strains of M . tuberculosis and the amount of proinflammatory {interleukin (IL)-8 and monocyte chemoattractant protein (MCP)- 1} and inhibitory (IL- 10) cytokines was measured in the culture supernatants by enzyme-linked immunosorbent assay (ELISA) . Infection with live bacilli induced de novo synthesis of IL-8, MCP-1 and IL-10, since cytokine release was abolished when cells were preincubated with the protein synthesis inhibitor cycloheximide . A differential production of antiinflammatory and inhibitory cytokines was observed . The amount of IL-8 and MCP-1 release was inversely related to strain virulence, the attenuated H37Ra strain being more prone than virulent strains to induce secretion of chemokines . In contrast, virulent strains induced greater amounts of the inhibitory cytokine IL-10 . Efficient upregulation of IL-10 synthesis, but not of chemokines, required infection of cells with live bacilli, since heat killing of organisms or challenge with soluble mycobacterial products completely abrogated the effect . Moreover, cells infected with virulent strains produced IL-10 even at a very low bacillus-to-cell ratio and secreted IL-10 continuously during the 96 h that followed infection . The results suggest that the degree of virulence affects host cell responses to M . tuberculosis infection . Continued production of IL-10 may be one of the means by which M . tuberculosis downregulates acute local inflammatory reactions, favoring the development of tuberculosis.

Bangladesh Med Res Counc Bull, 2000 Dec, 26(3), 87 - 91
Detection of Mycobacterium leprae by Polymerase Chain Reaction; Haque AK et al.; The improved procedure based on Polymerase Chain Reaction (PCR) for detection of M . leprae has been developed . The sensitivity and specificity of this method were tested using different concentration of genomic DNA of M . leprae Thai 53 and genomic DNAs from mycobacterial species and related microorganisms respectively . Application of this method to biopsy samples obtained from Bangladesh was conducted and detected M . leprae DNA in 7 of the 10 clinical specimens . Acid fast bacilli were not detected in four of the seven positive cases under the microscopic observation . It was concluded that this method was sensitive and specific for detection of M . leprae in clinical specimens and also simple to detect in only one step of PCR.

Int J Syst Evol Microbiol, 2001 Nov, 51(Pt 6), 1949 - 57
Legionella gresilensis sp . nov . and Legionella beliardensis sp . nov., isolated from water in France; Lo Presti F et al.; Novel Legionella-like isolates, strains Montbeliard A1T and Greoux 11 D13T, isolated from two different French water sources, were studied taxonomically and phylogenetically . Morphological and biochemical characterization revealed that they were Gram-negative, aerobic, non-spore-forming bacilli with a cut-glass appearance that grew only on L-cysteine-supplemented buffered charcoal yeast extract agar . Phenotypic characterization using fatty acid and ubiquinone profiles and SDS-PAGE analysis confirmed that they were closely related, but distinct from, other species of the genus Legionella, since serotyping could not relate them to any existing serogroup . Genotypic profiles generated by randomly amplified polymorphic DNA and 16S-23S rDNA spacer region PCR analyses were unique for each of these isolates . DNA-DNA relatedness values of strains Montbeliard A1T and Greoux 11 D13T to each other and to other Legionella type strains were less than 25% . Phylogenetic affiliation of these organisms obtained by 16S rDNA sequence comparisons confirmed that they were distinct from any other known Legionella species . All the above results confirm that these strains constitute two novel species for which the names Legionella gresilensis sp . nov . (type strain Greoux 11 D13T = ATCC 700509T = CIP 106631T) and Legionella beliardensis sp . nov . (type strain Montbeliard A1T = ATCC 700512T = CIP 106632T) are proposed.

Indian J Pediatr, 2001 Oct, 68(10), 931 - 5
Pediatric sarcoidosis in India; Gupta SK; OBJECTIVE: Since 1957, when the first pediatric case of sarcoidosis was reported, 11 more cases have been traced in the Indian literature . METHODS: Nine of them were reported from general wards of hospitals (while the remaining 3 were from pediatric unit of AIIMS, New Delhi) . Failure of initial treatment with anti-tuberculosis drugs for some months, necessitated search for an alternative diagnosis . Considerable delay (several months to years) occurred due to several parent- or physician-dependant factors . RESULTS: Affection more in girls (9 to 12), universal fever and constitutional symptoms, loss of weight, scanty lung features, hepatomegaly, often with massive splenomegaly, frequent lymphadenopathy etc . caused initial confusion . CONCLUSION: Treatment with oral steroid or with chloquine and NSAIDS with or without steroid MDI gave equally good results . Long follow-up was done in a few cases only, showing relapses in nearly 66% . One case had a superinfection with acid-fast bacilli.

Am J Ophthalmol, 2002 Jan, 133(1), 144 - 5
Bilateral endogenous endophthalmitis caused by HACEK microorganism; Sullivan P et al.; PURPOSE: To report a case of bilateral endogenous endophthalmitis caused by a HACEK group organism (Actinobacillus actinomycetemcomitans) . METHODS: An 85-year-old Caucasian female presented with bilateral light perception visual acuity, anterior chamber cells, hypopyon, posterior synechiae, and no red reflex . RESULTS: The patient underwent bilateral vitrectomy, with topical, intravitreal, and systemic antibiotics . Vitreous cultures revealed gram-negative cocco bacilli consistent with Actinobacillus actinomycetemcomitans . At final follow-up, visual acuity was 20/25 RE and 20/40 LE . CONCLUSIONS: Prompt diagnosis and treatment of HACEK {Hemophilus sp . (parainfluenzae, aphrophilus, paraphrophilus), Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corodens, and Kingella kingae} group bacterial endophthalmitis can yield good results.

Am J Respir Crit Care Med, 2001 Dec 15, 164(12), 2213 - 9
Iron-cofactored superoxide dismutase inhibits host responses to Mycobacterium tuberculosis; Edwards KM et al.; Superoxide dismutase (SOD) is a ubiquitous metalloenzyme in aerobic organisms that catalyzes the conversion of superoxide anion to hydrogen peroxide . Mycobacterium tuberculosis is unusual in that it secretes large quantities of iron-cofactored SOD . To determine the role of SOD in pathogenesis, we constructed mutants of M . tuberculosis H37Rv with reduced SOD production . Compared with controls, SOD-diminished isolates were more susceptible to killing by hydrogen peroxide . The isolates were markedly attenuated, exhibiting nearly 100,000-fold fewer bacilli than virulent control strains in the lungs and spleens of C57BL/6 mice 4 wk after intravenous inoculation . In the lung, SOD-attenuated M . tuberculosis induced robust interstitial mononuclear cell infiltration within 24 h and many cells were apoptotic by TUNEL staining, whereas virulent H37Rv exhibited minimal early inflammatory response and only rare interstitial mononuclear cell apoptosis . During prolonged infections, C57BL/6 mice tolerated SOD-attenuated M . tuberculosis better than BCG, exhibiting 68% greater weight gain, quicker eradication of bacilli from the spleen, and less alveolar lung infiltration . These results establish the importance of SOD in the pathogenesis of tuberculosis . Its effect appears to be mediated in part by inhibiting innate host immune responses, including early mononuclear cell infiltration of infected tissues and apoptosis.

Infect Immun, 2002 Jan, 70(1), 292 - 302
DNA vaccine combinations expressing either tissue plasminogen activator signal sequence fusion proteins or ubiquitin-conjugated antigens induce sustained protective immunity in a mouse model of pulmonary tuberculosis; Delogu G et al.; DNA vaccination has emerged as a powerful approach in the search for a more efficacious vaccine against tuberculosis . In this study, we evaluated the effectiveness of immunizing with combinations of 10 different tuberculosis DNA vaccines that expressed mycobacterial proteins fused at the N terminus to eukaryotic intracellular targeting sequences . In one vaccine combination, the genes were fused to the tissue plasminogen activator signal sequence (TPA), while in a second combination the same 10 genes were expressed as ubiquitin (Ub)-conjugated proteins . In ex vivo studies in which the secretion of gamma interferon was measured, cellular immune responses were detected in mice vaccinated with either the TPA DNA vaccine combination or the Ub DNA vaccine combination at 7 and 14 days following a low-dose Mycobacterium tuberculosis challenge . Moreover, mice vaccinated with the TPA combination, the Ub combination, and Mycobacterium bovis BCG were able to limit the growth of tubercle bacilli in the lung and spleen after a virulent tuberculous aerosol challenge . Histopathological analyses also showed that mice immunized with the DNA vaccine combinations had substantially improved postinfection lung pathology relative to the naive controls . Finally, in three different long-term experiments, the survival periods following aerogenic challenge were extended as much as sevenfold for vaccinated mice compared to naive controls . Interestingly, in all three experiments, no significant differences were detected in the mean times to death for mice immunized with the TPA combination or the Ub combination relative to the BCG controls . In conclusion, these studies demonstrate the effectiveness of immunization with DNA vaccine combinations against tuberculosis and suggest that further testing of these plasmid cocktails is warranted.

Infect Immun, 2002 Jan, 70(1), 286 - 91
Dependence of Mycobacterium bovis BCG on anaerobic nitrate reductase for persistence is tissue specific; Fritz C et al.; Mycobacterium bovis BCG, the only presently available vaccine against tuberculosis, was obtained from virulent M . bovis after serial passages in vitro . The vaccine strain retained at least some of its original virulence, as it persists in immune-competent hosts and occasionally may cause fatal disease in immune-deficient hosts . Mycobacterial persistence in vivo is thought to depend on anaerobic metabolism, an apparent paradox since all mycobacteria are obligate aerobes . Here we report that M . bovis BCG lacking anaerobic nitrate reductase (NarGHJI), an enzyme essential for nitrate respiration, failed to persist in the lungs, liver, and kidneys of immune-competent (BALB/c) mice . In immune-deficient (SCID) mice, however, bacilli caused chronic infection despite disruption of narG, even if growth of the mutant was severely impaired in lungs, liver, and kidneys . Persistence and growth of BCG in the spleens of either mouse strain appeared largely unaffected by lack of anaerobic nitrate reductase, indicating that the role of the enzyme in pathogenesis is tissue specific . These data suggest first that anaerobic nitrate reduction is essential for metabolism of M . bovis BCG in immune-competent but not immune-deficient mice and second that its role in mycobacterial disease is tissue specific, both of which are observations with important implications for pathogenesis of mycobacteria and vaccine development.

Ann Diagn Pathol, 2001 Dec, 5(6), 350 - 3
Mycobacterium avium intracellulare otitis media; Trupiano JK et al.; Atypical mycobacterial infections of the middle ear are extremely rare . To our knowledge, only eight cases have been reported in the literature, five of which involve Mycobacterium avium intracellulare . We present a case of culture-proven, M . avium intracellulare otomastoiditis in an 8-year-old boy with common variable immunodeficiency syndrome . The patient clinically presented with pain and otorrhea . The histopathology was marked by acid-fast bacilli-laden histiocytes . Consideration of this entity in the differential diagnosis of chronic, recalcitrant otorrhea can lead to timely diagnosis, treatment, an decreased morbidity .

Ann Thorac Cardiovasc Surg, 2001 Oct, 7(5), 301 - 3
Asymptomatic atresia of the lobar bronchus of the lung: a case report; Suemitsu R et al.; A 64-year old woman presented with an asymptomatic occlusion of the intermediate bronchus associated with a peripheral mass occupying the entire middle and lower lobes . As malignancy was suspected, inferior bilobectomy was done . There was a complete atelectasis of both lobes, with massive parenchymal necrosis . Pathological examinations suggested a tuberculous granuloma in the bronchus and parenchyma although tuberculous bacilli were not found . This case was unusual as congenital anomaly, and was suspected as bronchial tuberculosis.

Spine, 2001 Dec 15, 26(24), E565 - 70
Tuberculous spondylitis: analysis of 69 cases from Saudi Arabia; Alothman A et al.; DESIGN: Retrospective chart and radiographic review of all cases admitted to the authors' hospital and diagnosed to have tuberculous spondylitis . OBJECTIVE: To identify the frequency, presenting features, utility of diagnostic tests, and treatment outcome of spinal tuberculosis in a tertiary care center . METHODS: All cases of spinal tuberculosis treated in the authors' hospital over 14 years were reviewed . Only those with histologic or microbiologic confirmation or those who responded to antituberculous spondylitis therapy were included . Outcome was assessed according to predefined criteria . RESULTS: Between 1985 and 1998, 69 cases of tuberculous spondylitis were found, i.e., 5% of all tuberculous spondylitis cases diagnosed . The mean age was 52.8 years (range, 15-80 years), and 37 (53.6%) were male . Only five patients had a history of tuberculosis . The most frequent symptoms were backache (84%) and fever (32%), and the most frequent sign was spinal tenderness (45%) . Nineteen (28%) patients had paraparesis and 12 (17%) had kyphosis . Except for elevated sedimentation rate (94.5%), the laboratory workup was not helpful . Both computed tomography and magnetic resonance imaging were found to be helpful in diagnosis, often with complementary information . Thirty-eight patients (55%) had thoracic spine involvement and 55 patients (80%) had paraspinal abscesses . Tissue aspirates had a yield of 70%, 35%, and 57% for granulomas, acid-fast bacilli smear, and culture, respectively . Medical therapy alone was given in 37 cases (54%), whereas 32 patients (46%) required additional surgical intervention, mainly those with spinal cord compression, spinal deformity, or risk of spinal instability . There was definite improvement in 63 cases (91%) . The best outcome was in those patients presenting early before the occurrence of spinal deformity or neurologic symptoms . CONCLUSION: Tuberculous spondylitis is prevalent in the authors' hospital . Both computed tomography and magnetic resonance imaging are extremely helpful for diagnosis, and tissue aspirate is a good confirmatory method . A good outcome is expected if the diagnosis is made in early stages before the appearance of spinal deformity and neurologic symptoms . Surgical intervention can be avoided in these cases regardless of the presence or absence of paraspinal abscesses.

Int J Antimicrob Agents, 2001 Dec, 18(6), 579 - 82
In vitro activity of the chelating agents nitroxoline and oxine against Mycobacterium bovis BCG; Murugasu-Oei B et al.; The chelating antibiotic nitroxoline (5-nitro-8-hydroxyquinoline) showed a bacteriostatic effect at a concentration of 10 microM for Mycobacterium bovis BCG . At higher concentrations the compound showed moderate cidal activity against growing bacilli . In contrast, its non-nitrated derivative oxine (8-hydroxyquinoline, MIC=2 microM) reduced the viability of a growing culture rapidly, 5000-fold at a concentration where the nitroxoline was merely bacteriostatic . Both compounds showed appreciable cidal activity against bacilli in their hypoxic dormant state.

Int J Antimicrob Agents, 2001 Dec, 18(6), 525 - 30
Efficacy of macrolides used in combination with ethambutol, with or without other drugs, against Mycobacterium avium within human macrophages; Schiavano GF et al.; The activities of clarithromycin or roxithromicin used in combination with other antimicrobial drugs were tested in human macrophages experimentally infected with 23 strains of Mycobacterium avium . Overall, clarithromycin-ethambutol-rifampicin was the most active combination tested . The reduction in intracellular viable bacilli was found to be more than 1 log(10) for 95% and more than 2 logs(10) for 65% of the strains . The second most active combination was roxithromycin-ethambutol-rifampicin, which was found to be bactericidal for about 80% and highly bactericidal for 20% of the strains . Others combinations were only bacteriostatic or weakly bactericidal for many of the strains . The addition of a third drug did not necessarily promote enhanced bacterial killing inside the macrophage.

Pediatr Dermatol, 2001 Sep-Oct, 18(5), 393 - 5
Isolation of drug-resistant tubercle bacilli in cutaneous tuberculosis; Ramesh V et al.; Cutaneous tuberculosis in a teenage boy that did not respond to conventional antitubercular therapy is described . Mycobacterium tuberculosis was recovered on culture and found to be resistant to rifampicin and isoniazid . Gradual regression of the lesions was observed after the addition of streptomycin and ethambutol . Later streptomycin was stopped and the boy was advised to continue ethambutol along with ofloxacin and thioacetazone until complete regression was achieved.

Pediatr Dermatol, 2001 Sep-Oct, 18(5), 384 - 7
Lichenoid sarcoidosis in a 3-year-old girl; Seo SK et al.; We report a case of lichenoid sarcoidosis in a 3-year-old girl . She had numerous discrete skin-colored or erythematous, infiltrated follicular papules on the buttocks and extremities since 2 months of age . Histopathologic examination showed follicular plugging and an upper dermal granulomatous infiltrate of epithelioid cells closely surrounding the follicular ducts . No acid-fast bacilli were seen in the sections examined . Chest radiograph and high-resolution computed tomography (CT) showed no hilar lymphadenopathy or pulmonary parenchymal changes . An angiotensin-converting enzyme level was elevated . The Mantoux reaction was negative and results of ophthalmologic examinations were normal . Treatment was started with triamcinolone 0.2 mg/kg and prednicarbate ointment . Some lesions healed completely and others showed residual pitting.

Pediatr Infect Dis J, 2001 Sep, 20(9), 912 - 4
Parotid abscess caused by Mycobacterium tuberculosis; Chatterjee A et al.; Tuberculosis of the parotid gland is rare . A 16-month-old US-born male infant with immigrant parents from Sudan presented to his primary care physician with periorbital cellulitis and preauricular lymphadenitis . He underwent incision and drainage of an abscess in the right intraparotid lymph node . The aspirate was positive for acid-fast bacilli by auramine-rhodamine stain and subsequently grew Mycobacterium tuberculosis . Antitubercular medications were started postoperatively.

Pediatr Infect Dis J, 2001 Nov, 20(11), 1061 - 5
Use of purified protein derivative to assess the risk of infection in children in close contact with adults with tuberculosis in a population with high Calmette-Guérin bacillus coverage; Almeida LM et al.; SETTING: Household contacts <15 years of age of adults with tuberculosis (TB) attending a reference center in Aracaju, Sergipe, Northeast Brazil . OBJECTIVE: To assess the use of purified protein derivative (PPD) and frequency of infection in children with high Calmette-Guerin bacillus (BCG) coverage who were recently exposed to TB . DESIGN: Cross-sectional study of 141 exposed household contacts <15 years of age and 506 nonexposed neighborhood controls . Children were examined and assessed for degree of exposure to index cases, BCG vaccination and scar and were tested with PPD . RESULTS: Exposed children were younger and less up to date in their vaccination schedule than controls (P < 0.05) . BCG had been given to 95.6% of exposed children and 97.4% of controls, but only 80.9% of exposed vs . 88.5% of controls had a scar (P < 0.05) . Scar sizes of exposed children were smaller (medians, 4.5 and 7 mm, respectively; P < 0.05) . Children had lower weight for age z scores and height than the National Center for Health Statistics standards . Exposed children had lower weight for age z scores than controls (P < 0.05) . Sixty-seven (47.5%) exposed children and 18 (3.6%) controls had PPD readings of >10 mm . Positivity and induration sizes increased with age, although this was significant only in the controls . The presence of a BCG scar was not associated with having a positive PPD . The degree of exposure was an important factor for PPD positivity; 66 (60.6%) of the 109 children with close exposure were positive compared with 1 (3.1%) of 32 with less intimate exposure . PPD indurations among close contacts were also larger than those with restricted exposure and controls (16.3, 11 and 9.4 mm, respectively; P < 0.05) . PPD positivity was associated with the quantification of acid-fast bacilli in the adult; 38.5% of contacts with adults with sputum with (+) were positive, compared with 42.3% of those with (++) and 58.4% with (+++) (P < 0.05) . CONCLUSION: PPD is a useful test to identify children infected with TB independently of whether they had received BCG or not . Children exposed to adults with TB are at high risk of infection . The risk of infection is associated with the intimacy of contact and the number of bacilli expectorated in sputum.

J Antimicrob Chemother, 2001 Dec, 48(6), 877 - 80
The early bactericidal activity of a low-clearance liposomal amikacin in pulmonary tuberculosis; Donald PR et al.; The early bactericidal activity (EBA) of a liposomal preparation of amikacin (MiKasome) with a long plasma half-life of 120-200 h was examined in seven patients with newly diagnosed, smear-positive pulmonary tuberculosis . Liposomal amikacin was given in slow iv infusions of 30 mg total amikacin/kg body weight on three successive days . Cfu counts were set up on 16 h sputum collections preceding the first dose and following each dose and were used for calculating the EBA . Despite the high concentrations of total amikacin, >1000 mg/L, obtainable in plasma, no evidence of EBA was obtained . In view of the considerable activity of liposomal amikacin in experimental murine tuberculosis, this finding indicates that liberation of amikacin from the long-life liposomes occurs only in macrophages that are not usually present in the vicinity of the large extracellular clumps of bacilli in the cavity caseum.

J Antimicrob Chemother, 2001 Dec, 48(6), 869 - 76
The activity of low-clearance liposomal amikacin in experimental murine tuberculosis; Dhillon J et al.; Most of the amikacin in low-clearance liposomal amikacin is excreted very slowly, offering the possibility of maintaining effective treatment of pulmonary tuberculosis with widely separated supervised doses . As a preliminary to explorations in humans, its efficacy was assessed in acute experimental murine tuberculosis by weekly counts of viable bacilli in spleen and lungs over a 4 week period . Liposomal amikacin in dosages of 160, 80 and 40 mg/kg given iv three times a week was 2.4-5.0 times more active than free amikacin and 6.6-6.7 times more active than streptomycin with the non-liposomal drugs given im five times a week . When the free amikacin and the streptomycin were also given iv three times a week, liposomal amikacin was 2.7-2.9 times more active than free amikacin and 3.7-5.6 more active than streptomycin . In a model of chronic tuberculosis, initial BCG vaccination was followed by challenge with virulent Mycobacterium tuberculosis and a 2 week stabilization period . Thereafter, treatment with liposomal amikacin 160 and 80 mg/kg three times a week for the first 4 weeks and then once a week for a further 4 weeks, had greater initial bactericidal activity than free amikacin 160 mg/kg five times a week, but had less eventual sterilizing activity than five times a week oral isoniazid 25 mg/kg or rifampicin 15 mg/kg . Although low-clearance liposomes increased the safety, potency and dosing interval of amikacin in these models, all aminoglycosides, including liposomal amikacin, were only bactericidal in the presence of bacillary metabolism and growth.

Nihon Kokyuki Gakkai Zasshi, 2001 Sep, 39(9), 699 - 704
{A case of endobronchial tuberculosis associated with bronchial asthma treated with high doses of inhaled corticosteroid}; Suzuki K et al.; A 46-year-old woman had been treated with 1,600-2,000 micrograms/day of beclomethasone dipropionate (BDP) and oral theophylline on the basis of a diagnosis of bronchial asthma in 1993 . Eosinophilic pneumonia was diagnosed in June 1999, and she was then treated with 40 mg/day of oral prednisolone (PSL), which was gradually tapered off, and then stopped in October 1999 . She was referred to our hospital because acid-fast bacilli were found in the sputum on January 18, 2000 . Her chest radiographs and CT scans showed partial atelectasis of the right upper lobe, and fiberoptic bronchoscopy revealed bronchial inflammatory changes and whitish mucosal nodular lesions in the walls of the lower trachea, the right main bronchus and the orifice of the right upper lobe bronchus . She was found to have endobronchial tuberculosis . Anti-tuberculosis treatment with isoniazid, rifampicin, streptomycin and pyrazinamide was started . Serum levels of interferon-gamma were markedly elevated on admission . Asthma symptoms improved for a period of one month after the beginning of anti-tuberculosis treatment, despite the termination of inhaled corticosteroid . However, as the tuberculosis improved, the frequency and severity of the asthma increased and so corticosteroid inhalation was started again . Four months after administration of the anti-tuberculosis drug, fiberoptic bronchoscopy revealed that the endobronchial lesions had improved without any stenosis or constrictive changes . It was speculated that high doses of inhaled corticosteroid may have the potential to cause endobronchial tuberculosis whilst, ironically, at the same time preventing bronchial stenosis by endobronchial tuberculosis . This is an interesting case in which the asthma symptoms first decreased during the acute phase of endobronchial tuberculosis and then increased again after the tuberculosis improved.

FEMS Microbiol Lett, 2001 Nov 27, 205(1), 113 - 7
Novel use of guanidinium isothiocyanate in the isolation of Mycobacterium tuberculosis DNA from clinical material; Chakravorty S et al.; Nucleic acid amplification technologies offer great promise for the rapid, sensitive and specific diagnosis of tuberculosis . However, the isolation of inhibitor-free DNA from biological specimens is a bottleneck of the PCR assay . Here we describe a simple method for the isolation of PCR-amplifiable DNA of Mycobacterium tuberculosis from all types of samples of pulmonary and extrapulmonary origin tested . Briefly, it involves concentration of the bacilli by high-speed centrifugation, removal of PCR inhibitors by a wash solution containing guanidinium isothiocyanate and the release of bacterial DNA by heating in the presence of detergents and Chelex-100 resin . The entire process is accomplished within approximately 3 h . The method has been validated on 780 samples of human, bovine and guinea pig origin including sputum, cerebrospinal fluid, pulmonary fluids, pus, fine needle aspirate, tissue, blood and milk.

Presse Med, 2001 Oct 27, 30(31 Pt 2), 11 - 5
{Chronic obstructive pulmonary disease and bronchial colonization/infection}; Fournier M; BACTERIAL FLORA IN THE SUBGLOTTAL AIRWAYS: In healthy non-smoking subjects, the subglottal airways are sterile . Inversely, bacteria are often isolated from the sublottal airways in patients with obstructive or non-obstructive chronic bronchitis, both during and between acute exacerbations . The significance of this bacterial colonization/infection, its natural history, and its impact on the course of chronic lung disease is poorly understood . BETWEEN EXACERBATIONS: 30 to 40% of all patients with chronic obstructive pulmonary disease in a stable situation without recent antibiotic therapy harbor potential pathogens in their intra-thoracic airways . The prevalence of Gram-negative bacilli increases in the more severe forms . This estimation is possibly biased due to the involuntary inclusion of patients with bronchial dilatation, since bronchectasia involves a high prevalence of bronchial colonization/infection . DURING EXACERBATIONS: The prevalence of bronchial colonization/infection by potential pathogens is to the order of 30-50%, irrespective of the bacteria isolated from endobronchial samples made during or between exacerbations . About half of the exacerbations would not be related to bacterial infection, but to viral infection . The other identified causes of exacerbations are: occupational or accidental exposure, co-morbidity (chronic sinus infection, chronic pharyngeal discharge, left ventricular failure, non-specific bronchial hyperreactivity) . INFLAMMATION: Inflammation of the bronchial mucosa is a constant feature of chronic obstructive pulmonary disease . The biological and histocytological features are distinctive from those observed in asthma . Polynuclear activation appears to play an important role in polynuclear infiltration of the mucosa, amplified during bacterial colonization.

Acta Derm Venereol, 2001 Aug-Sep, 81(4), 291 - 3
Cutaneous infection with Mycobacterium fortuitum after localized microinjections (mesotherapy) treated successfully with a triple drug regimen; Nagore E et al.; Mesotherapy is a treatment method devised for controlling pain syndromes or diseases by subcutaneous microinjections given at or around the involved areas at short intervals of time . Different adverse effects have been described due to this modality of treatment . This report describes 3 patients with cutaneous infection caused by Mycobacterium fortuitum after mesotherapy . Three women, aged 24, 27 and 44 years, presented with similar clinical features, consisting of painful nodules located at the points where mesotherapy had been applied . A smear from a skin biopsy revealed the presence of acid-fast bacilli in all 3 cases . The specimen was cultured and eventually identified as M . fortuitum . A multidrug long-term regimen (combinations of 3 drugs from the following: ciprofloxacin, cotrimoxazole, clarithromycin and amoxicillin-clavulanic acid) was needed to achieve resolution of the lesions . After 15, 25 and 26 months of follow-up, no patient relapsed . Mycobacterium fortuitum is a rapidly growing mycobacterium that can lead to cutaneous infection after minor surgical procedures when aseptic measures are not adequate . Multiple drugs for several months are usually needed to treat this disease successfully.

Life Sci, 2001 Nov 2, 69(24), 2899 - 910
A novel emulsifier, labrasol, enhances gastrointestinal absorption of gentamicin; Hu Z et al.; Gentamicin (GM) is an important aminoglycoside antibiotic for the treatment of infections caused by a wide spectrum of aerobic gram-negative bacilli and gram-positive cocci . As a class, the aminoglycosides are poorly absorbed from the gastrointestinal (GI) tract and are commonly used as injectable and topical preparations . This study was aimed at finding the effect of a novel emulsifier, Labrasol, on the absorption of GM from the GI tract of rats . GM formulations were prepared, either as saline solution or as Labrasol microemulsions, and were administrated to rat small intestine and colon . Plasma GM levels following intestinal application were compared to those obtained with intravenous (i.v.) administration . A 5 mg/kg dose of GM preparation containing Labrasol, 1 ml/kg, administrated into colon resulted in the mean AUC of 21.179+/-1.374 microg x h/ml, compared to 7.813+/-0.105 microg x h /ml obtained with i.v . administration of GM, 1 mg/kg . The absolute bioavailability (BA) of the Labrasol preparation was 54.2% . Labrasol facilitates the transmucosal delivery of GM from rat colon by forming microemulsions, and the BA obtained with Labrasol microemulsion was higher than with other surfactants (8.4% for Tween 80 and 3.4% for Transcutol P) . Additionally, in vitro permeation studies demonstrated that Labrasol also inhibited the intestinal secretory transport . The effect of Labrasol is ascribed to both (1) enhanced GM absorption from the GI lumen into the systemic circulation and (2) inhibition of efflux of GM from the enterocytes to the GI lumen.

BMC Microbiol . 2001;1(1):26 . Epub 2001 Oct 17.
Identification of two Mycobacterium tuberculosis H37Rv ORFs involved in resistance to killing by human macrophages; Miller BH et al.; BACKGROUND: The ability of Mycobacterium tuberculosis to survive and replicate in macrophages is crucial for the mycobacterium's ability to infect the host and cause tuberculosis . To identify Mycobacterium tuberculosis genes involved in survival in macrophages, a library of non-pathogenic Mycobacterium smegmatis bacteria, each carrying an individual integrated cosmid containing M . tuberculosis H37Rv genomic DNA, was passed through THP-1 human macrophages three times . RESULTS: Two of the clones recovered from this enrichment process, sur2 and sur3, exhibited significantly increased survival relative to wild-type bacteria . In coinfection experiments, the ratio of sur2 colonies to wild-type colonies was 1:1 at 0 hours but increased to 20:1 at 24 hours post phagocytosis . The ratio of sur3 colonies to wild-type colonies was 1:1 at 0 hours and 5:1 at 24 hours . The M . tuberculosis ORFs responsible for increased survival were shown to be Rv0365c for the sur2 clone and Rv2235 for the sur3 clone . These ORFs encode proteins with as-of-yet unknown functions . CONCLUSIONS: We identified two M . tuberculosis ORFs which may be involved in the ability of tubercle bacilli to survive in macrophages.

J Exp Med, 2001 Nov 19, 194(10), 1421 - 32
Processing of Mycobacterium tuberculosis antigen 85B involves intraphagosomal formation of peptide-major histocompatibility complex II complexes and is inhibited by live bacilli that decrease phagosome maturation; Ramachandra L et al.; Mycobacterium tuberculosis (MTB) inhibits phagosomal maturation to promote its survival inside macrophages . Control of MTB infection requires CD4 T cell responses and major histocompatibility complex (MHC) class II (MHC-II) processing of MTB antigens (Ags) . To investigate phagosomal processing of MTB Ags, phagosomes containing heat-killed (HK) or live MTB were purified from interferon-gamma (IFN-gamma)-activated macrophages by differential centrifugation and Percoll density gradient subcellular fractionation . Flow organellometry and Western blot analysis showed that MTB phagosomes acquired lysosome-associated membrane protein-1 (LAMP-1), MHC-II, and H2-DM . T hybridoma cells were used to detect MTB Ag 85B(241-256)-I-A(b) complexes in isolated phagosomes and other subcellular fractions . These complexes appeared initially (within 20 min) in phagosomes and subsequently (>20 min) on the plasma membrane, but never within late endocytic compartments . Macrophages processed HK MTB more rapidly and efficiently than live MTB; phagosomes containing live MTB expressed fewer Ag 85B(241-256)-I-A(b) complexes than phagosomes containing HK MTB . This is the first study of bacterial Ag processing to directly show that peptide-MHC-II complexes are formed within phagosomes and not after export of bacterial Ags from phagosomes to endocytic Ag processing compartments . Live MTB can alter phagosome maturation and decrease MHC-II Ag processing, providing a mechanism for MTB to evade immune surveillance and enhance its survival within the host.

Eur J Clin Microbiol Infect Dis, 2001 Sep, 20(9), 651 - 4
Gram-Negative anaerobic endocarditis: two case reports and review of the literature; Bisharat N et al.; The rarity of anaerobic gram-negative endocarditis limits the ability of physicians to define its prognosis . Two cases of endocarditis due to Bacteroidesfragilis are described, and a review of the English literature for all cases of anaerobic gram-negative endocarditis reported since 1940 is presented . The disease predominantly affects males . Clinical features are similar to those of endocarditis due to nonanaerobic organisms, but underlying heart disease is less common and the rate of thromboembolic complications is high . All deaths reported were due to Bacteroides spp.; no deaths due to Fusobacterium spp . have been reported . Treatment with metronidazole has dramatically improved the prognosis of patients with endocarditis due to anaerobic gram-negative bacilli.

Am J Clin Pathol, 2001 Nov, 116(5), 770 - 5
Amplified in situ hybridization with peptide nucleic acid probes for differentiation of Mycobacterium tuberculosis complex and nontuberculous Mycobacterium species on formalin-fixed, paraffin-embedded archival biopsy and autopsy samples; Zerbi P et al.; The aim of this study was to evaluate sensitivity and specificity of in situ hybridization (ISH) using peptide nucleic acid (PNA) probes and tyramide-based amplification for the differentiation between Mycobacterium tuberculosis (MTB) and mycobacteria other than tuberculosis (MOTT) on formalin-fixed, paraffin-embedded tissue samples . We performed ISH simultaneously with both probes on 86 specimens from different organs: 70 obtained at autopsy and 16 by biopsy, all with a histologic evidence of mycobacterial infection confirmed by Ziehl-Neelsen-positive staining . Taking culture as the "gold standard," the sensitivity and the specificity of the MTB probe were 100% (41/41) and 95% (38/40), respectively . In only 2 cases ISH failed to identify mycobacteria . Culture results were not available in 3 cases . We propose ISH as a relatively simple and rapid method to differentiate mycobacteria on formalin-fixed, paraffin-embedded specimens (it is more specific than usual histologic stains) and as an alternative to polymerase chain reaction, allowing the morphologic evaluation of positive bacilli.

Acta Histochem, 2001 Oct, 103(4), 433 - 6
Leprosy in Israel: an imported disease--the support of histopathological examination for its detection; Lejbkowicz F et al.; Leprosy is rare and non-endemic in Israel . Cases of leprosy are invariably imported by immigrants or foreign workers arriving from endemic areas . In view of the relative rarity of the disease, clinicians and pathologists are not always alert to the possibility of the disease or recognize potential symptoms . A case history is presented of a 31-year-old immigrant presenting symptoms of skin lesions and nodules on the hands and facial region, especially the ear lobe . Confirmation of the infection was provided by histopathology of suspected lesions stained for acid-fast bacilli (modified Fite-Faraco staining).

Arch Pathol Lab Med, 2001 Nov, 125(11), 1491 - 3
Granulomatous colitis associated with botryomycosis of Propionibacterium acnes; Wada R et al.; Propionibacterium acnes, an anaerobic, non-spore-forming, gram-positive bacillus, is a common inhabitant of the skin, and its virulence is considered to be low in humans . This report describes an unusual case of granulomatous colitis associated with P acnes infection in a 46-year-old woman . The affected cecum exhibited a tumor histologically characterized by massive transmural infiltrates of small lymphocytes and noncaseating epithelioid granulomas with multinucleated giant cells . Botryomycotic granules were also found in the muscular layer and paracolic connective tissues and consisted of gram-positive bacilli with filamentous growth . Polymerase chain reaction confirmed the presence of P acnes 16S ribosomal DNA in the surgical specimen of the colon . The patient developed a postoperative P acnes-induced peritonitis, which subsided with treatment with antibiotics and surgical drainage . The present case indicates that P acnes is one of the possible pathogens for granulomatous colitis.

Pediatrics, 2001 Nov, 108(5), 1143 - 8
Molecular epidemiology of antibiotic-resistant gram-negative bacilli in a neonatal intensive care unit during a nonoutbreak period; Toltzis P et al.; OBJECTIVE: Gram-negative organisms that are resistant to parenteral antibiotics are a growing threat to hospitalized patients . This study was conducted to define the epidemiologic characteristics of these organisms during a nonoutbreak period in a neonatal intensive care unit (NICU) . METHODS: Nasopharyngeal and rectal swab specimens were obtained 3 times a week from every infant in a tertiary care NICU during a 12-month period . Specimens were processed to identify aerobic Gram-negative species resistant to gentamicin, piperacillin-tazobactam, or ceftazidime . Selected clinical parameters were tested for their association with colonization with a resistant organism . Restriction endonuclease digests of genomic DNA were derived from isolates of the most frequently occurring species . The fragments were analyzed by pulsed-field gel electrophoresis (PFGE) to determine the genetic relatedness of the various isolates and thereby determine the length of colonization, the frequency of horizontal transmission, and the size and duration of clusters . RESULTS: A total of 101 infants (8.6%) of 1180 admissions were colonized with at least 1 antibiotic-resistant bacillus before NICU discharge . Multiple parameters indicating a prolonged, complicated NICU course were associated with resistant colonization, including gestational age, length of stay, and exposure to several classes of antibiotics . Colonization with resistant bacilli occurred as early as the first NICU day, but acquisition continued throughout the infants' stay . A total of 436 isolates were analyzed by PFGE . On the basis of this molecular analysis, it was determined that duration of colonization was usually very short; the median for all species tested was <1 week . In addition, cross-colonization occurred in only 12% of all PFGE-analyzed isolates . Most clusters of cross-colonized infants were small, with the majority involving only 2 patients . CONCLUSIONS: During endemic periods, acquisition of antibiotic-resistant Gram-negative bacilli in the NICU may occur very soon after admission, but colonization continues over many weeks of NICU stay . The duration of colonization with resistant bacilli is short, and horizontal transmission is unusual . These characteristics suggest a gradual but temporary incorporation of these organisms from the NICU environment into the nascent newborn microflora over time with little cross-colonization . These observations may aid the rational development of infection-control strategies to contain the reservoir of resistant Gram-negative organisms in the NICU.antibiotic resistance, Gram-negative bacilli, neonatal intensive care, antibiotic utilization, colonization, pulsed-field gel electrophoresis.

Mikrobiol Z, 2001 Jul-Aug, 63(4), 9 - 14
{New strains of soil bacilli which mineralize organic phosphorus compounds}; Roi AA et al.; Microorganisms, which mineralize organophosphates (216 isolates) have been isolated for the chernozem soils of different regions of Ukraine . Sixteen most active strains referred to Bacillus genus have been identified and studied . These microorganisms grow (with different intensity) on the mineral medium with glucose and glycerophosphate or phytin, accumulating phosphate-ions in the medium . The most active strains mineralized glycerophosphate, accumulating about 280 micrograms/ml of PO4(3-) and phytin accumulating up to 130 micrograms/ml of PO4(3-) during 3 days . New strains of bacilli have been chosen for creation of combined bacterial preparations of the complex effect.

Diagn Microbiol Infect Dis, 2001 Sep-Oct, 41(1-2), 57 - 63
Molecular mechanisms of cefoxitin resistance in Escherichia coli from the Toronto area hospitals; Forward KR et al.; Escherichia coli may become resistant to cephamycines and oxyimino cephalosporins by virtue of promotor and attenuator mutations or because they have acquired mobilized beta-lactamases from other gram-negative bacilli . This study examined Canadian strains to determine how often promotor and/or attenuator mutations account for this mechanism of resistance and the extent to which clonal spread of these organisms has occurred . We sequenced the promotor and attenuator region of 30 strains resistant to cefoxitin . Twenty-two strains had promotor mutations, 26 had attenuator mutations . Most promotor mutations resulted either in a change in the -35 promotor region towards the E . coli sigma 70 consensus sequence or in the creation of a new consensus hexamer upstream . Eight strains had mutations that increased the typical ampC 16-nucleotide spacer region to the consensus 17- or an 18-nucleotide sequence . Of the attenuator mutations, most did not substantially affect the attenuator loop . Several of the mutations have previously been described in South Africa, Scandinavia, and France . There was evidence that strains bearing certain mutations were clonally disseminated; however, the 11 strains bearing a complex set of attenuator mutations were not . The majority of cephamycin resistant E . coli strains in Toronto have attenuator and/or promotor mutations upstream of the chromosomal ampC gene.

J Clin Microbiol, 2001 Nov, 39(11), 4131 - 7
Detection of rifampin resistance in Mycobacterium tuberculosis in a single tube with molecular beacons; El-Hajj HH et al.; Current clinical assays for determining antibiotic susceptibility in Mycobacterium tuberculosis require many weeks to complete due to the slow growth of the bacilli . Here we demonstrate an extremely sensitive single-tube PCR assay that takes less than 3 h and reliably identifies rifampin-resistant M . tuberculosis in DNA extracted directly from sputum . Ninety-five percent of mutations associated with rifampin resistance occur in an 81-bp core region of the bacterial RNA polymerase gene, rpoB . All mutations that occur within this region result in rifampin resistance . The assay uses novel nucleic acid hybridization probes called molecular beacons . Five different probes are used in the same reaction, each perfectly complementary to a different target sequence within the rpoB gene of rifampin-susceptible bacilli and each labeled with a differently colored fluorophore . Together, their target sequences encompass the entire core region . The generation of all five fluorescent colors during PCR amplification indicates that rifampin-susceptible M . tuberculosis is present . The presence of any mutation in the core region prevents the binding of one of the molecular beacons, resulting in the absence of one of the five fluorescent colors . When 148 M . tuberculosis clinical isolates of known susceptibility to rifampin were tested, mutations associated with rifampin resistance were detected in 63 of the 65 rifampin-resistant isolates, and no mutations were found in any of the 83 rifampin-susceptible isolates . When DNA extracted directly from the sputum of 11 patients infected with rifampin-resistant tuberculosis was tested, mutations were detected in all of the samples . The use of this rapid assay should enable early detection and treatment of drug-resistant tuberculosis in clinical settings.

Eur J Clin Microbiol Infect Dis, 2001 Aug, 20(8), 576 - 80
Two cases of pyomyositis caused by Klebsiella pneumoniae and review of the literature; Wang TK et al.; Two patients with diabetes mellitus developed Klebsiella pneumoniae pyomyositis involving multiple muscles and other metastatic foci associated with bacteraemia . The cases are presented here, and the 22 cases of gram-negative pyomyositis reported previously in the literature are reviewed . As gram-positive cocci cause 99% of pyomyositis, it is postulated that the pathogenesis of pyomyositis may be associated with the ability of the bacteria to adhere to muscles by various cell surface adhesins that are not present in gram-negative bacilli . It is also postulated that a high serum glucose concentration may facilitate the growth and formation of the Klebsiella pneumoniae capsule, hence increasing its virulence and causing serious disseminated Klebsiella infections in diabetic patients.

J Formos Med Assoc, 2001 Aug, 100(8), 568 - 70
Tuberculous synovitis of the elbow joint; Lin YM et al.; Tuberculous synovitis in the elbow joint is extremely rare in developed countries . We describe a 68-year-old man who had had a gradually enlarging mass over the volar side of the left proximal forearm near the elbow joint for 4 months . Plain roentgenograms of the diseased elbow showed early osteoarthritic change . Magnetic resonance imaging revealed diffuse synovitis with a large 8 x 8 cm extra-articular synovial cyst . Synovectomy was performed and histopathologic examination of the surgical specimen revealed granulomatous inflammation with caseation, prominent Langhan's giant cells, and sparse acid-fast bacilli . The patient had been receiving antituberculous chemotherapy for at least 8 months at the time of examination and had no recurrence of swelling or discharging sinuses during follow-up . Differential diagnoses in patients with elbow swelling should include pigmented villonodular synovitis, hemophilic arthropathy, rheumatoid arthritis, degenerative joint disease, and tuberculosis . Simple aspiration may enable earlier diagnosis, before destructive arthropathy becomes advanced.

J Mol Evol, 2001 Dec, 53(6), 680 - 9
Mycobacterium tuberculosis phylogeny reconstruction based on combined numerical analysis with IS1081, IS6110, VNTR, and DR-based spoligotyping suggests the existence of two new phylogeographical clades; Sola C et al.; This paper deals with phylogenetic relationships among a set of 90 clinical strains representative of the worldwide diversity of the Mycobacterium tuberculosis complex (Kremer et al . 1999) using eight independent genetic markers: IS6110, IS1081, the direct repeat (DR) locus, and five variable number of tandem DNA repeat loci (VNTR) . In a preliminary experiment, phylogenetic trees based on single markers were constructed that led to the detection of some similarities between the VNTR-based and the spoligotyping-based phylogenetic trees . In the second step, a more global phenetic approach based on pairwise comparison of strains within each typing system was used, followed by calculations of mean genetic distances based on all the eight loci and the use of the neighbor-joining algorithm for tree reconstruction . This analysis confirmed our preliminary observations and suggested the existence of at least two new phylogeographical clades of M . tuberculosis, one defined as the "East African-Indian family" (EA-I), which may find its origin on the African or Asian continents, and the other as the "Latin American and Mediterranean" (LA-M) family . The existence of these two families was also validated by an independent phylogenetic analysis of spoligotyping on a larger set of shared types (n = 252) and further corroborated by VNTR and katG-gyrA results . The potential origin of these families of bacilli is discussed based on cattle domestication and human migration history . In conclusion, the information contained in insertion sequence and repetitive DNAs may serve as a model for the phylogenetic reconstruction of the M . tuberculosis complex.

Proc Natl Acad Sci U S A, 2001 Oct 23, 98(22), 12706 - 11
Mycobacterium tuberculosis signal transduction system required for persistent infections; Zahrt TC et al.; It is estimated that nearly 2 billion people currently suffer from latent Mycobacterium tuberculosis infection . Although the key front-line antituberculosis drugs are effective in treating individuals with acute tuberculosis, these drugs are ineffective in eliminating M . tuberculosis during the persistent stages of latent infection . Consequently, therapeutics that directly target persistent bacilli are urgently needed . We have conducted a global analysis on a group of regulatory determinants that may play a role in M . tuberculosis virulence, and identified a two-component response regulator whose expression is required for entrance into and maintenance of persistent infection . Inactivation of this response regulator, Rv0981 (termed here mprA for mycobacterial persistence regulator), affected M . tuberculosis H37Rv growth in vivo in an organ- and infection stage-specific fashion . These results indicate that two-component systems are important for adaptation of the tubercle bacillus during stages of persistent infection.

Int J Antimicrob Agents, 2001 Sep, 18(3), 217 - 22
In vitro susceptibilities of Bartonella and Rickettsia spp . to fluoroquinolone antibiotics as determined by immunofluorescent antibody analysis of infected Vero cell monolayers; Ives TJ et al.; The in vitro susceptibilities of Bartonella and Rickettsia spp . to different concentrations of ciprofloxacin, levofloxacin, ofloxacin and sparfloxacin in Vero cell cultures, were determined by enumeration of immunofluorescent-stained bacilli . After incubation in a CO(2)-enriched atmosphere, inocula were replaced and tested with media containing 12 different concentrations of each antibiotic in replicate for each species and the monolayers were re-incubated . Growth status was determined by evaluation of immunofluorescent staining bacilli . Effective inhibitory antibiotic dilution endpoints were determined by counting Bartonella- and Rickettsia-specific fluorescent foci across a range of antibiotic dilutions with an epi-fluorescent microscope, and were compared with an antibiotic-negative control . Based upon the use of C(max):MIC and AUC:MIC data, levofloxacin exhibited activity against Bartonella elizabethae and B . quintana.

Ophthal Plast Reconstr Surg, 2001 Sep, 17(5), 374 - 80
Orbital lipogranulomatous inflammation harboring Mycobacterium abscessus; Gonzalez-Fernandez F et al.; PURPOSE: A case of iatrogenic Mycobacterium abscessus orbital infection is presented to call attention to its distinct histopathologic appearance, mechanism of organism, virulence, and clinical management . METHODS: Clinicopathologic analysis of an orbital infection caused by M . abscessus is described . Detailed histologic analysis is performed to provide insight into the mechanism of infection and correlate the features of the orbital infection with that of atypical mycobacteriosis at other body sites . RESULTS: A 71-year-old woman had an orbital mass in the supranasal orbit after a blepharoplasty . The mass consisted of a dimorphic inflammatory reaction with a superficial purulent reaction and a deeper granulomatous process consisting of epithelioid tubercles, each centered about a lipid vacuole . Acid-fast bacilli were found in the lipid vacuoles but not elsewhere in the specimen . Each tubercle was surrounded by a zone of lymphocytes and a desmoplastic reaction . Microbiologic culture studies identified M . abscessus . A combination of surgical intervention and antibiotic therapy for 4 weeks eliminated the infection . CONCLUSIONS: Acid-fast stains should be performed on any orbital lesion showing an apparent lipogranulomatous reaction . Although clinical management of orbital atypical microbacteriosis is difficult, the combination of surgical and specific antimicrobial intervention is effective . Our study contributes to an evolving understanding of the mechanism of human infectivity of these low-virulence organisms by suggesting that the orbital fat is a source of lipid material that can harbor the organisms, allowing them to escape host immunosurveillance.

Hist Sci Med, 1998 Sep, 32(3), 297 - 300
{Tholozan and plague in Persia}; Mollaret HH; In Persia since 1858, Tholozan studied between 1870 and 1882 the plague foci of the iranian Kurdistan which shall be dealt a century later (1947-1963) with Dr . M . Baltazard and his co-workers from the Pasteur Institute of Teheran . Tholozan had already pointed out the localization of the disease in some well defined villages and gave a good clinical description mentioning the traces of flea bites on the patients skin . One knows nowadays that wild rodents (Meriones) are the storing places of the plague bacilli in the Kurdistan . Tholozan's observations confirmed by modern ones allow to consider him a great loimologist of modern times.

Int J Tuberc Lung Dis, 2001 Oct, 5(10), 963 - 7
The prognosis of respiratory failure in patients with tuberculous destroyed lung; Park JH et al.; SETTING: The medical intensive care unit of a tertiary referral hospital . OBJECTIVE: To determine the prognosis of patients whose lungs are damaged by previous and/or present tuberculosis infection and who have subsequently been presented with acute respiratory failure requiring mechanical ventilation . DESIGN: A consecutive series of 38 patient cases with retrospective data analysis . RESULTS: Pulmonary function test results for tests performed within the previous year were made available in 21 of the 38 cases (55%) . These showed a mean (+/- SD) forced vital capacity (FVC) of 1.52 +/- 0.46 L (41.0 +/- 14.5% predicted), a forced expiratory volume/second (FEV1) of 0.77 +/- 0.18 L (29.3 +/- 13.6% predicted), and an FEV1/FVC ratio of 55.1 +/- 16.2% . The acid-fast bacilli (AFB) positive group had a significantly higher mortality and more severe lung destruction when compared with the AFB-negative group . Patients with positive AFB were significantly more hypocapnic than those with negative AFB (6.4 +/- 2.7 vs . 9.3 +/- 3.9 kPa, P = 0.020) . In multivariate analysis, the level of PaCO2 on admission was identified as the only significant prognostic index (OR 0.76, 95%CI 0.60-0.96) . CONCLUSION: Patients with positive AFB smears or cultures may have higher mortality rates than those with negative AFB in the tuberculosis destroyed lung patients with acute respiratory failure . A higher PaCO2 measurement could indicate a better survival rate in this group of patients.

Acta Otolaryngol, 2000 Mar, 120(2), 279 - 85
Experimental intravenous inoculation of Klebsiella rhinoscleromatis bacilli in albino rats: a histopathological and bacteriological study; Gaafar HA et al.; Scleroma, chronic specific granuloma of the nose and upper respiratory tract, is endemic in Egypt and many other countries . The exact pathogenesis of the disease as regards the aetiological role of Klebsiella rhinoscleromatis is contradictory . This work investigated the effect of experimental intravenous injection of K . rhinoscleromatis in albino rats to demonstrate that the micro-organism can fulfil Koch's postulates . Micro-organisms were isolated from biopsy specimens taken from nasal lesions of 10 patients in the granulomatous stage of scleroma . Specimens were subjected to bacteriological and histopathological examinations to confirm the diagnosis . A 100 microl volume of freshly prepared bacterial inoculum containing 10(8) cfu/ml was injected weekly in the tail vein of each of 30 albino rats for 5 consecutive weeks . Biopsy specimens were taken from sacrificed animals and subjected to bacteriological and histopathological examinations . Positive histopathological diagnosis of scleroma was reported in the nose of 66.7% of rats, the larynx of 46.7%, the lungs of 26.7% and liver of 20% of rats . Bacteriological techniques were successful in revealing K . rhinoscleromatis from the nose of 36.7% of rats, the larynx of 30% and the lungs of 20% of rats . Various techniques were carried out to demonstrate the micro-organisms in tissue sections . Two histochemical stains for bacteria were employed: silver and Periodic Acid Schiff (PAS) stains . Immunoperoxidase technique using Klebsiella capsular type 3 antiserum was applied . It gave positive results in 66.7% of the 6 stained liver sections in spite of negative bacteriological cultures . The histiocytic nature of the Mikulicz cells was confirmed using alpha-1 antitrypsin, an immunohistochemical marker of histiocytes, and by studying the ultrastructural features of Mikulicz cells using the transmission electron microscope.

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3109 - 12
Activities of new macrolides and fluoroquinolones against Mycobacterium ulcerans infection in mice; Bentoucha A et al.; Mice infected in the left hind footpad with 5 log(10) acid-fast bacilli of Mycobacterium ulcerans were divided into an untreated control group and 17 treatment groups that received one of the following regimens for 4 weeks (all doses in milligrams per kilogram): 100 mg of azithromycin (AZM), 100 mg of clarithromycin (CLR), or 50 mg of AZM for a duration of 5 days a week (daily), three times a week, or once weekly . In addition, the following regimens were administered daily: 100 mg of telithromycin (TLM), sparfloxacin (SPX), or moxifloxacin (MOX); 200 mg of levofloxacin (LVX); 100 mg of streptomycin (STR) or amikacin (AMK); 10 mg of rifampin (RIF); and the combination of 10 mg of RIF and 100 mg of AMK (RIF+AMK) . After completion of treatment, mice were observed for 30 weeks . The effectiveness of treatment regimens was assessed in terms of the delay in median time to footpad swelling in treated mice compared with that in the untreated controls . Clear-cut bactericidal activity, i.e., an observed delay in footpad swelling that exceeded the period of treatment, was observed in the STR-, AMK-, and RIF+AMK-treated mice . However, all mice treated with either AMK or STR alone had swollen footpads before the end of the 30-week observation period, suggesting regrowth of M . ulcerans . In contrast, 50% of the mice treated with the RIF+AMK combination exhibited no lesion even after 30 weeks, suggesting cure . The remaining regimens could be assigned to one of three groups: (i) no activity (50 mg of AZM, 100 mg of AZM thrice weekly, TLM, and LVX); (ii) bacteriostatic activity, i.e., a delay in footpad swelling shorter than the 4-week treatment duration (100 mg of AZM daily or once weekly, CLR thrice or once weekly, and MOX); or (iii) weak bactericidal activity (CLR daily and SPX) . The RIF+AMK combination and possibly RIF+STR warrant further study for the treatment of M . ulcerans infection in humans.

Singapore Med J, 2001 Jul, 42(7), 325 - 7
Paradoxical enlargement of tuberculous brain abscess during drug treatment: a case report; Ng SK et al.; This is a case report of a patient suffering from tuberculous (TB) brain abscesses . Despite supervised and appropriate anti-TB chemotherapy, the size of the abscesses paradoxically increased, accompanied by clinical deterioration at 1 month . A second aspiration was performed which yielded a large amount of viable tubercle bacilli . The same drug regimen was continued . This was followed by complete resolution of the lesions in 12 months . Hence TB brain abscess which remains culture and smear positive after four weeks treatment may not represent treatment failure.

Rev Invest Clin, 2001 Jul-Aug, 53(4), 315 - 23
{Drug resistance of Mycobacterium tuberculosis in Orizaba, Veracruz . Implications for the tuberculosis prevention and control program}; Garcia-Garcia ML et al.; BACKGROUND: Tuberculosis, declared a global emergency by the World Health Organization, continues to be an important public health problem in Mexico, included in the first twenty causes of death . OBJECTIVE: To know the impact of drug resistance of Mycobacterium tuberculosis on treatment outcome, need of re-treatment and mortality in a cohort of patients with pulmonary tuberculosis receiving directly observed therapy, short course (DOTS) . METHODS: We conducted a population-based study in a suburban region in Southern Mexico . People who had been coughing for more than two weeks underwent sputum acid-fast bacilli smear . Patients with a positive smear were recruited and underwent clinical exam, chest X-ray, HIV testing, and sputum cultures . Identification, drug susceptibility testing and restriction fragment length polymorphism analysis (RFLP) were performed in all isolates . Patients were followed every 12 months for new episodes of tuberculosis and vital status . Patients were referred for clinical care to the local program of tuberculosis . Deaths were corroborated with death certificates . Informed consent was obtained from participants . RESULTS: Between March 1995 and February 1999, tuberculosis was diagnosed in 371 patients who were followed for an average of 32 months . M . tuberculosis was cultured from 316 patients; resistance to any drug occurred in 25.0% of isolates (primary 18.8%, acquired 49.2%); only to isoniazid in 6.8% (primary 7.3%, acquired 4.8%); to isoniazid and rifampin in 6.2% (primary 1.6%, acquired 23.8%) . Patients with drug resistance had a higher probability of treatment failure (OR = 16.9, CI 95% 4.5-63.0) and patients with MDR strains had a higher probability of need of re-treatment (RR = 24.4, CI 95% 8.8-67.6), and of death (RR = 4.0, CI 95% 1.5-10.7) . Additional variables were found to be associated with subsequent episodes of disease and mortality: Cocaine use, chronic disease, type of radiological lesions, HIV co-infection, non-compliance and treatment delay, as well as RFLP clustering . CONCLUSIONS: In this study, we observed that drug resistance showed a severe impact on the outcome and survival; drug-resistance was the most significant factor for these negative outcomes; DOTS may not be sufficient in areas where drug resistance is considerable, and patient follow-up for longer periods of time, as compared to evaluation at the end of treatment, provides additional information which is useful for prevention and control programs.

Dtsch Tierarztl Wochenschr, 2001 Sep, 108(9), 386 - 9
Cilia-Associated Respiratory (CAR) bacillus infection in veal calves and adult cattle; Pravettoni D et al.; Cilia-Associated Respiratory (CAR) bacillus is a filamentous bacterium that colonizes the ciliated epithelium of the respiratory tract of many animal species and that has been associated with chronic inflammatory lesions in naturally and experimentally infected rats, mice and rabbits . In the present study, the prevalence of CAR bacillus infection and histological lesions of the trachea in veal calf and adult cattle were investigated . Forty five healthy veal calves and 45 adult cattle, raised in 18 different herds were selected at slaughter . From each animal, a tracheal sample was processed for histology, stain-ed with the Warthin-Starry method to evaluate the presence of CAR bacillus, and with haematoxylin and eosin to evaluate the presence of inflammatory lesions . CAR bacillus was identified in 17 veal calves (37.7%) and in 7 adult cattle (15.5%) . Inflammatory lesions were found in 42 veal calves (93.3%) and in 41 adult cattle (91.1%) . Statistical analysis pointed out a significant correlation between the presence and number of CAR bacilli and the presence and number of lymphoid follicles (P = 0.0071) and the presence and severity of neutrophilic infiltrates (P = 0.0428) . These results indicate that CAR bacillus infection is common in cattle and is correlated with tracheal inflammatory lesions.

J Perinatol, 2001 Sep, 21(6), 376 - 81
Purulence and gram-negative bacilli in tracheal aspirates of mechanically ventilated very low birth weight infants; Cordero L et al.; OBJECTIVE: Tracheal aspirates (TAs) from mechanically ventilated very low birth weight (VLBW) infants are frequently obtained during the evaluation of suspected sepsis, tracheitis, or ventilator-associated pneumonia (VAP) . Purulence and bacteria in Gram stain of bronchopulmonary secretions are considered signs of respiratory infection, and medical decisions are made on the assumption that they are predictors of positive bacterial tracheal cultures (TCs) . The purpose of this retrospective investigation was to establish the relationship of purulence and bacteria in TA from ventilated VLBW infants with positive TC and to identify its clinical significance . STUDY DESIGN: One hundred and seventy consecutively born VLBW infants (1996 to 1998) who remained on mechanical ventilation longer than 1 week were studied . Demographic, laboratory, and clinical data were obtained from hospital medical records . Purulence, defined by the number of polymorphonuclear leukocytes (PMNs) per low power field (LPF), was reported as light (<25 PMNs/LPF) or moderate/heavy (>or=25 PMNs/LPF) for every TA . RESULTS: Purulence was absent in 469 of 646 (72%) TA taken from 170 infants . Light purulence was present in 17% and moderate/heavy purulence in 11% . TCs were positive in 58% of non-purulent, 94% of light, and 100% of moderate/heavy purulent TA . Bacteria on Gram stain were present in 12% of non-purulent, 70% of light purulent, and 83% of moderate/heavy purulent TA . Moderate/heavy purulence in TA was predictive of a positive TC with Gram-negative bacilli (GNB) with 70% sensitivity, 100% specificity, 100% positive predictive value, and 67% negative predictive value . Purulence in TA, as well as GNB airway colonization, became more frequent as mechanical ventilation progressed and was not associated with a particular GNB species . There were 79 infants who never had purulent TA and 91 who, at some time during the hospitalization, did . At the time of first purulent TA, 65 (71%) of 91 infants were asymptomatic . Twenty-six infants (29%) had clinical deterioration for which they underwent sepsis work-up . Three had blood stream infection, 5 VAP, 5 tracheitis, and 13 respiratory complications of non-infectious etiology . Four of five VAP infants died; all others survived . CONCLUSION: In VLBW infants, purulence in TA is associated with prolonged endotracheal intubation and is temporally related to GNB airway colonization . At the time of the first purulent TA, the majority of mechanically ventilated VLBW infants are asymptomatic . Only a few symptomatic VLBW infants had nosocomial respiratory infection . Understanding the clinical significance of purulence and GNB in TA from this unique patient population is important for management and prognosis, and it may decrease concern for infection and the associated use of antibiotics.

Rev Pneumol Clin, 2001 Sep, 57(4), 289 - 95
{Active tracheo-bronchial caseous tuberculosis . Ten patients}; Le Huu L et al.; BACKGROUND: We report 10 cases of active endobronchial tuberculosis in female patients.PATIENTS AND METHODS: Diagnosis of endobronchial tuberculosis was established by demonstrating the presence of Koch bacilli in the bronchial washings or at pathology examination of a bronchial mucosa biopsy . Follow-up was available for 5 of the 10 cases.RESULTS: These women were aged 19 to 756 years . Clinical signs included persistent cough (n=10), wheezing (n=10), dyspnea (n=5), dysphonia (n=4) and fever (n=8) . Chest x-ray was normal in 4 cases but showed condensation in 4 others with infiltration of the superior lobe in 2 cases . Bronchoscopy findings were characteristic with a thick layer of white caseum on the mucosa . Koch bacilli were found in all bronchial wash specimens . Pathology examination confirmed the diagnosis of tuberculosis in 5 of the 10 cases.

J Postgrad Med, 2001 Jan-Mar, 47(1), 30 - 2
Pulmonary nocardiosis in human immunodeficiency virus infection: a tuberculosis mimic; Subhash HS et al.; Patients with human immunodeficiency virus (HIV) infection are prone to develop pulmonary infections like nocardiosis . It is often misdiagnosed as pulmonary tuberculosis since the manifestations are similar . A twenty-seven years old male presented with fever, cough with expectoration and weight loss for two months . Chest radiograph showed opacity in the right mid zones . Sputum smears were negative for acid fast bacilli (AFB) and revealed gram positive branching filamentous organisms resembling Nocardia species . Subsequently, Nocardia was grown on sputum culture . HIV antibody was positive by ELISA test . He was treated with co-trimoxazole . If sputum is repeatedly tested negative for AFB in the setting of radiological suspicion of tuberculosis, testing for Nocardia species should be considered in the HIV-infected patients.

Rev Neurol, 2001 Aug 16-31, 33(4), 325 - 8
{Tuberculous meningioma en plaque without extracerebral tuberculosis . A case report}; Casadevall-Codina T et al.; INTRODUCTION: Tuberculosis continues to be an intensly extended disease in under developed countries in immunodepressed and immunocompetent persons . In the central nervous system tuberculous lesions are the most frequent cause of space occupying lesions . CLINICAL CASE: We report the case of a young, immunocompetent woman from Guinea, currently living in Spain . Following a seizure, neuro imaging showed there to be a right frontal space occupying lesion . The characteristics of the MR images obtained were fundamental to the determination of the meningeal site of the lesion en plaque and to suggest the aetiology as being an infectious inflammatory process, although a meningioma could not be ruled out . The absence of signs of extracerebral disease, the localization and distribution of the lesion and lack of short term response to medical treatment, which covered the different diagnostic possibilities, made it necessary to do a diagnostic meningeal biopsy . CONCLUSIONS: MR imaging is a great help in the diagnosis of tuberculous lesions of the central nervous system . The differential diagnosis between tuberculous pachymeningitis and meningioma, both in the form of plaques, is extremely difficult without obtaining a biopsy specimen . The finding of epithelioid granulomas with caseous necrosis and/or acid alcohol fast bacilli in the biopsy specimen is diagnostic of tuberculosis . A single meningeal lesion situated in the cortex but with no associated extracerebral disease causes considerable difficulty in diagnosis . Tuberculosis must always be remembered in view of the increasing number of immigrants from under developed countries.

Ophthalmology, 2001 Oct, 108(10), 1819 - 25
Clinical profile and outcome in Bacillus endophthalmitis; Das T et al.; OBJECTIVE: To study the clinical presentation, microscopic and organismal culture correlation of vitreous, and species-specific outcome in Bacillus endophthalmitis DESIGN: Retrospective noncomparative case series . INTERVENTION: Thirty-one culture proven Bacillus endophthalmitis patients between January 1991 and February 1998 underwent vitrectomy and intravitreal antibiotic injection . Lensectomy was combined when considered essential, and on a few occasions intravitreal dexamethasone was added . The patients also received topical and systemic antibiotics . The undiluted vitreous biopsy was the source for microbiologic evaluation (microscopy and culture sensitivity) . MAIN OUTCOME MEASURES: The duration of symptoms, the presenting visual acuity, and influence of intravitreal dexamethasone with intravitreal antibiotics were examined for any statistical correlation with the final visual acuity . RESULTS: Trauma was the major cause of infection . Vitreous biopsy microscopy demonstrated gram-positive bacillus in 28 of 31 cases, and polymicrobial infection was seen in 12 instances . All Bacillus species were sensitive to gentamicin, followed by vancomycin and ciprofloxacin . Clinical treatment within 7 days of symptoms, use of intravitreal vancomycin, and absence of polymicrobial infection were associated with better visual outcome . CONCLUSIONS: With appropriate treatment that essentially consists of vitrectomy and intravitreal antibiotics, patients with Bacillus endophthalmitis are likely to benefit in many instances . Gram-positive bacilli detected on vitreous microscopy should be empirically treated as Bacillus species unless otherwise proved.

Nihon Hansenbyo Gakkai Zasshi, 2001 Aug, 70(3), 151 - 5
{Chemotherapy of leprosy: theoretical basis of new guideline in Japan}; Goto M; For the effective treatment of leprosy, we should consider that (1) more time is needed for the elimination of bacilli than ordinary bacterial infection, (2) bactericidal therapy often induces host immunity called reactions, (3) rapid treatment is needed for the reactions . Last year, ad hoc committee of Japanese Leprosy Association recommends standard treatment protocol of leprosy in Japan, which is a modification of World Health Organization's multidrug therapy . For multibacillary(MB) with bacterial index (BI > or = 3) before treatment, 2 years treatment by rifampicin, dapsone and clofazimine (MDT/MB) is necessary . When BI decrease is not satisfactory(BI value decrease < 2 steps, or final BI > or = 3) after 2 years, MDT/MB should be continued until BI negativity and loss of active lesions . Theoretical background of our proposal is described.

AIDS, 2001 Sep 28, 15(14), 1875 - 9
Integration of tuberculosis screening at an HIV voluntary counselling and testing centre in Haiti; Burgess AL et al.; OBJECTIVE: To describe the integration of tuberculosis screening into the activities of an HIV voluntary counselling and testing (VCT) centre in a country with endemic tuberculosis . SETTING: An HIV VCT centre in Port au Prince, Haiti . DESIGN: All patients presenting for HIV VCT who reported cough received same-day evaluation for active tuberculosis . Of the 1327 adults presenting to the centre for the first time between January and April 1997, 263 (20%) reported cough and of these 241 (92%) were evaluated . RESULTS: Of the 241 patients evaluated for cough, 76 (32%) were diagnosed with pulmonary tuberculosis . Of the 76 patients diagnosed with pulmonary tuberculosis, 28 (37%) had a positive smear for acid-fast bacilli (AFB), 14 (18%) had a negative AFB smear but a positive sputum culture for Mycobacterium tuberculosis, and 34 (45%) had culture-negative tuberculosis . Also, 31 out of 241 (13%) VCT clients evaluated for cough were diagnosed with bacterial pneumonia . CONCLUSION: This report confirms that in areas with a high HIV and tuberculosis prevalence, a high proportion of VCT clients have active pulmonary tuberculosis . The integration of tuberculosis screening offers several benefits, including the diagnosis and treatment of large numbers of individuals with tuberculosis, a decreased risk of nosocomial tuberculosis transmission, and the opportunity to provide tuberculosis prophylaxis to HIV-positive patients in whom tuberculosis has been excluded . Future studies are needed to determine the cost-effectiveness of integrated tuberculosis and HIV VCT services, and whether integration should be recommended in all countries with high HIV and tuberculosis rates.

Neth J Med, 2001 Oct, 59(4), 177 - 80
Detection of Mycobacterium tuberculosis complex DNA in pericardial fluid, bone marrow and peripheral blood in a patient with pericardial tuberculosis . A case report; Tzoanopoulos D et al.; Definitive diagnosis of tuberculous pericarditis requires identification of bacilli in pericardial fluid or tissue . Conventional diagnostic methods are time-consuming and have a low sensitivity making bacteriological confirmation of the disease very difficult . Hereby, we report the case of molecular detection of Mycobacterium tuberculosis in pericardial fluid, bone marrow and peripheral blood from a 63-year-old woman with pericardial tuberculosis, using a nested PCR assay specific for IS6110 insertion element of M . tuberculosis complex . The patient had an excellent response to a three-drug combination anti-tuberculous regimen and 1 year later was asymptomatic, without evidence of constrictive pericarditis.

J Infect Dis, 2001 Oct 15, 184(8), 1082 - 5 Epub 2001 Sep 28.
Granulysin-dependent killing of intracellular and extracellular Mycobacterium tuberculosis by Vgamma9/Vdelta2 T lymphocytes; Dieli F et al.; Contribution of Vgamma9/Vdelta2 T lymphocytes to immune protection against Mycobacterium tuberculosis is still a matter of debate . It was reported earlier that Vgamma9/Vdelta2 T lymphocytes kill macrophages harboring live M . tuberculosis through a granule-dependent mechanism that results in killing of intracellular bacilli . This study found that Vgamma9/Vdelta2 T lymphocytes reduce the viability of both extracellular and intracellular M . tuberculosis . Granulysin and perforin, both detected in Vgamma9/Vdelta2 T lymphocytes, play a major role, which indicates that Vgamma9/Vdelta2 T lymphocytes directly contribute to a protective host response against M . tuberculosis infection.

J Clin Microbiol, 2001 Oct, 39(10), 3718 - 20
Rapid detection of Mycobacterium tuberculosis in contaminated BACTEC 12B broth cultures by testing with Amplified Mycobacterium Tuberculosis Direct Test; Zheng X et al.; Contamination of broth cultures of acid-fast bacilli (AFB) by bacterial species other than Mycobacterium species frequently occurs . Many of these contaminated cultures require redecontamination and reincubation before the appropriate tests can be performed for identification, significantly affecting the turnaround time for reporting culture results . In this study, the Amplified Mycobacterium Tuberculosis Direct Test (MTD; Gen-Probe) was performed to detect the Mycobacterium tuberculosis complex (MTBC) in 125 BACTEC 12B broth cultures with positive growth indices . Among these, 41 grew non-AFB bacteria only, and all 41 were negative by the MTD . The remaining 84 bottles contained contaminated cultures that grew both AFB and other bacteria or yeasts . Repeat decontamination and reincubation of these specimens required a mean time of 13 days (range, 3 to 40 days) . The MTD results were positive for 10 samples, 9 of which were MTBC culture positive and 1 of which grew Myobacterium celatum, a species known to cross-react in the MTD . All cultures growing other mycobacterial species were negative by the MTD . The results of this study demonstrate that the MTD is both sensitive and specific in detecting MTBC in contaminated broth cultures and that, when used selectively, the MTD can potentially rule in or out a diagnosis of MTBC as much as 12 days earlier than using nonamplified DNA probe testing alone can.

Eur J Cardiothorac Surg, 2001 Oct, 20(4), 722 - 7
Surgical intervention for pulmonary tuberculosis: analysis of indications and perioperative data relating to diagnostic and therapeutic resections; Furak J et al.; OBJECTIVE: To analyze the data on patients operated on for pulmonary tuberculosis (TB) with (Group I) or without (Group II) a correct TB diagnosis and preoperative anti-TB treatment . METHODS: Between 1980 and 1997, 144 resections for TB (Groups I+II) were performed . The 80 patients in Group I underwent therapeutic resections: 32 cases involved recurrent cavities or tuberculomas, three involved post-TB bronchiectasis, 13 involved progression of cavities or tuberculomas, and 32 involved persistent tuberculomas after 6 months of anti-TB therapy . The 64 patients in Group II were operated on for a suspicion of malignancy in 49 cases, for cavitary lesions with haemophthysis in six cases, for multiple lesions in seven cases, and for recurrent hydrothorax in two cases . RESULTS: Groups I and II included 0 and five pneumonectomies, 32 and 29 lobectomies, 48 and 20 wedge resections, 0 and nine videothoracoscopic biopsies, and 0 and one hilar lymphadenectomy, respectively . In Groups I and II, the mean duration of postoperative hospitalization was 13.2 and 10.4 days, and the frequency of postoperative pneumothorax was 11.25 and 4.6%, respectively . The incidence of bronchopleural fistula was 1.25 and 0%, the mortality was 0 and 3.1%, and the morbidity was 53.7 and 35.9% in Groups I and II, respectively . Two patients with active disease died in Group II . Pathology demonstrated that the frequency of acid-fast bacilli in Groups I and II was 40 and 25%, respectively . CONCLUSIONS: Patients without a correct preoperative TB diagnosis underwent more extensive parenchyma resection . Postoperative complications increased when acid-fast bacilli were present . The lack of preoperative anti-TB treatment did not involve a higher risk of minor complications, but death occurred only in this group.

Int J Tuberc Lung Dis, 2001 Sep, 5(9), 855 - 60
Yield of smear, culture and amplification tests from repeated sputum induction for the diagnosis of pulmonary tuberculosis; Al Zahrani K et al.; OBJECTIVES: To assess the yield of repeated sputum induction for the diagnosis of active tuberculosis in patients who do not produce spontaneous sputum, or with smear-negative spontaneous samples . METHODS: Induced sputum was examined with fluorescent microscopy, two amplification methods (PCR Amplicor MTB, and MTD2), and cultured for mycobacteria using liquid (Bactec 12B) and Lowenstein-Jensen media . Bronchoscopy and collection of other specimens were performed at the discretion of the treating physician . RESULTS: A total of 1115 sputum inductions performed in 500 patients without adverse events yielded an adequate specimen in 1113 (99.8%), and microbiological confirmation in 43 of 44 (98%) culture-positive active TB cases . Yield increased with repeated sputum induction . The cumulative yield for acid-fast bacilli smear and mycobacterial culture was 64% and 70% respectively for one, 81% and 91% for two, 91% and 99% for three, and 98% and 100% for four induced samples . Yield of PCR also increased with the greater number of induced samples tested . CONCLUSIONS: Repeated sputum induction could considerably improve diagnostic accuracy for pulmonary TB.

Avian Dis, 2001 Jul-Sep, 45(3), 755 - 9
Tuberculosis in geese (Anser anser) in Turkey; Ozcan K et al.; Tuberculosis was detected in seven geese at the breeding unit of the Faculty of Veterinary Medicine, Kafkas University, during 1998 . In the necropsy, lesions of nodular type were seen in the liver, spleen, and lungs . Histopathologically, the lesions were characterized by central areas of caseous necrosis surrounded by epithelioid cells, multinucleated giant cells, lymphocytes, and an outer fibrous capsule . Acid-fast bacilli were visualized by the Ziehl-Neelsen staining method in paraffin sections and smears . Inoculation into Lowenstein-Jensen media with glycerin yielded Mycobacterium spp.

Avian Dis, 2001 Jul-Sep, 45(3), 709 - 18
Experimental inoculation of European starlings (Sturnus vulgaris) and American crows (Corvus brachyrhynchos) with Mycobacterium bovis; Butler KL et al.; The purpose of this series of pilot studies was to determine whether the passerine species studied are susceptible to infection with Mycobacterium bovis . Separate experiments were conducted on wild-caught starlings (Sturnus vulgaris) and American crows (Corvus brachyrhynchos) . In each experiment, four birds were challenged intraperitoneally and four were challenged orally with microorganisms . Challenge dose was 1 x 10(5) colony-forming units of M . bovis cultured from a white-tailed deer (Odocoileus virginianus) case in Michigan . Birds were euthanatized at 1 and 2 mo postinoculation . Histologic lesions suggestive of mycobacteriosis, without the presence of acid-fast bacilli, were noted in all experimental groups . Mycobacterial cultures performed on pooled tissue samples were positive for M . bovis in only some of the intraperitoneal inoculates of each species.

Aesthetic Plast Surg, 2001 Jul-Aug, 25(4), 295 - 8
Upper eyelid mycobacterial infection following Oriental blepharoplasty in a pulmonary tuberculosis patient; Chen SH et al.; Bilateral upper blepharoplasty for the Oriental eyelid was performed in a 20-year-old female on antituberculous therapy for 3 months . The postoperative course was smooth and the patient was back to normal life . But unfortunately, at 3 months after the upper blepharoplasty, a spherical tumescence and red granuloma developed over the right upper eyelid . The granuloma was resected, and on pathological examination the specimen revealed epitheloid granuloma with Langhan's giant cells and a few acid-fast positive bacilli . The clinical events and pathological findings were suggestive of Mycobacterium tuberculosis rather than Mycobacterium chelonei as the possible cause of infection . Thus the patient was advised to continue antituberculous therapy and no antibiotic was prescribed . The eyelid swelling resolved gradually and was completely normal at the end of antituberculous therapy . Though it has been suggested that aesthetic surgery can be performed safely 3 months after antituberculous therapy in a patient with pulmonary tuberculosis, the remote risk of such a complication is always a possibility.

Semin Respir Infect, 2001 Sep, 16(3), 169 - 76
Extended-spectrum beta-lactamases; Blondeau JM; Extended-spectrum beta lactamase (ESBL) producing gram-negative bacilli are a growing concern, especially because the species of organisms producing these enzymes are increasing . Bacteria possessing these enzymes are resistant to third-generation cephalosporins--antimicrobial agents important for inpatient therapy . These resistant organisms are clinically important because they result in increased morbidity and mortality . Additionally, some laboratories may have difficulty detecting ESBL-producing organisms . These and other issues are discussed in this article .

Southeast Asian J Trop Med Public Health, 2001 Jun, 32(2), 390 - 3
Improved method of direct microscopy for detection of acid-fast bacilli in sputum; Aung WW et al.; Microscopy of direct smears for acid-fast bacilli (AFB) is the most commonly used method for diagnosis of tuberculosis . However, direct microscopy of sputum, though rapid, has low sensitivity and there is a need for improved methods . Sputum samples were collected from patients attending the Union Tuberculosis Institute, Yangon . The microscopy of smears made directly from sputum were compared with the microscopy after liquefaction of sputum with household bleach (NaOCl) and concentration of bacteria by centrifugation . Out of 948 samples, 248 samples (26.2%) were positive for acid-fast bacilli by direct microscopy and 293 samples (30.9%) were positive for acid-fast bacilli by the household bleach method . There was a significant increase in the number of acid-fast bacilli positive samples by the house-hold bleach method (p<0.05) . The method is simple and cheap . As a disinfectant, household bleach has the advantage of lowering the risk of laboratory infection.

Rev Panam Salud Publica, 2001 Jun, 9(6), 368 - 74
{Prognostic factors for pulmonary tuberculosis outcome in Recife, Pernambuco, Brazil}; Albuquerque MF et al.; OBJECTIVE: The objective of this case-control study was to identify prognostic factors for the outcome of pulmonary tuberculosis among 297 patients who were treated between 1994 and 1999 at the Federal University of Pernambuco Clinics Hospital, in Recife, Pernambuco, Brazil . METHODS: The cases were defined as individuals whose treatment ended in death, dropout, or failure . The controls were persons whose treatment ended in cure . The following independent variables were submitted to uni- and multivariate analyses: sex, age, schooling, being a smoker, consuming alcohol, previous treatment for tuberculosis, response to the tuberculin test, HIV serology, resistance to antimicrobial agents, results of direct investigation of alcohol- and acid-resistant bacilli, and treatment approach used . After that, the uni- and multivariate analyses were repeated, including as cases only deaths and individuals with treatment failure . RESULTS: Four risk factors for tuberculosis treatment failure were found: excessive alcohol intake (odds ratio (OR) = 2.58; P = 0.014), co-infection with HIV (OR = 3.40; P = 0.028), previous tuberculosis treatment (OR = 4.89; P < 0.001), and resistance to two or more antituberculosis drugs (OR = 3.49; P = 0.017) . In the second multivariate analysis, which excluded dropout cases, no association was found between treatment outcome and excessive alcohol consumption, but the other associations remained . This result suggests a close relationship between alcoholism and treatment dropout . CONCLUSIONS: In the group that was studied the prognostic factors for failure of pulmonary tuberculosis treatment were interrelated and were of a biological, clinical, and social character . These factors should be identified at the beginning of treatment in order to allow implementation of specific follow-up procedures such as the strategy of directly observed treatment . This would strengthen tuberculosis control at the local level.

Respirology, 2001 Sep, 6(3), 217 - 24
Miliary tuberculosis: clinical manifestations, diagnosis and outcome in 38 adults; Mert A et al.; OBJECTIVE: The aim of the study was to determine the clinical, radiographic and laboratory characteristics, diagnostic methods, and prognostic variables in patients with miliary tuberculosis (TB) . METHODOLOGY: The records of 38 patients (15 male, 23 female; mean age 41 years, range 16-76 years) with miliary TB from 1978 to 1998 were analyzed . Patients were evaluated also as to whether they presented with a fever of unknown origin (FUO) . Criteria for the diagnosis of miliary TB were (i) miliary pattern on chest X-ray or (ii) biopsy or autopsy evidence of miliary organ involvement . Paraffin-embedded tissues with granulomata (n = 15) were re-evaluated for the presence of Mycobacterium tuberculosis DNA by polymerase chain reaction (PCR) . RESULTS: Predisposing conditions were present in 24% of the patients . The findings were fever, weakness, night sweats, anorexia/weight loss (100% for each), hepatomegaly (37%), splenomegaly (32%), choroidal tubercles (13%), neck stiffness (11%), altered mental status (8%), anaemia (76%), leukopenia (26%), thrombocytopenia (16%), lymphopenia (76%), pancytopenia (8%) and hypertransaminasemia (55%) . Eighteen patients (47%) met the criteria for a FUO . Miliary infiltrates were found on chest X-rays of 32 of 38 cases (84%) . In six cases without miliary infiltrates, the diagnosis was made by laparotomy in four cases, and autopsy in two cases . Tuberculin skin test was positive in 32% of cases . Acid-fast bacilli were demonstrated in 37% (16/43), and cultures for M . tuberculosis were positive in 90% (9/10) of tested specimens (predominantly sputum and bronchial lavage) . Granulomas were found in 85% (11/13) of lung, 100% (15/15) of liver, and 56% (9/16) of bone marrow tissue specimens . Acid-fast bacilli staining was negative in all (0/21), while PCR was positive in 47% (7/15) of specimens with granulomata . Mortality was 18% . Stepwise logistic regression identified male sex (P = 0.005), non-typical miliary pattern (P = 0.015), altered mental status (P = 0.002) and failure to treat for TB (P = 0.00001) as independent predictors of mortality . CONCLUSIONS: Miliary infiltrates on chest X-ray or FUO should raise the possibility of miliary TB . Therapy should be administered urgently to prevent an otherwise fatal outcome.

Am J Clin Pathol, 2001 Sep, 116(3), 347 - 53
Pulmonary granuloma caused by Pseudomonas andersonii sp nov; Han XY et al.; Pulmonary granuloma is a common lesion for which gram-negative bacteria are rarely implicated as a cause . Hence, most physicians are unaware of this etiology . We isolated a gram-negative bacterium from a surgically resected pulmonary granuloma in a 42-year-old, nonimmunocompromised woman . Within the necrotizing granuloma, numerous organisms also were demonstrated by Gram stain, suggesting a cause-disease relationship . Characterization of the bacterium by sequence analysis of the 16S ribosomal gene, cellular fatty acid profiling, and microbiologic studies revealed a novel bacterium with a close relationship to Pseudomonas . We propose a new species for the bacterium, Pseudomonas andersonii . These results suggest that the differential diagnosis of a lung granuloma also should include this gram-negative bacterium as a potential causative agent, in addition to the more common infections caused by acid-fast bacilli and fungi . This bacterium was shown to be susceptible to most antibiotics that are active against gram-negative bacteria.

Annu Rev Microbiol, 2001, 55, 139 - 63
Nonreplicating persistence of mycobacterium tuberculosis; Wayne LG et al.; There is ample clinical evidence, as well as evidence from animal experiments, that Mycobacterium tuberculosis can persist in tissues for months to decades without replicating, yet with the ability to resume growth and activate disease . Our knowledge of both macrophage physiology and the nature of tuberculous lesions in man and animals suggests that hypoxia is a major factor in inducing nonreplicating persistence (NRP) of tubercle bacilli . In vitro models reinforce this conclusion and provide insights into mechanisms that make NRP possible . There is evidence from in vitro models that the strategies employed by the bacilli to permit hypoxic NRP include restriction of biosynthetic activity to conserve energy, induction of alternative energy pathways, and stabilization of essential cell components to lessen the need for repair or replacement.

J Immunol, 2001 Sep 15, 167(6), 3308 - 15
ATP stimulates human macrophages to kill intracellular virulent Mycobacterium tuberculosis via calcium-dependent phagosome-lysosome fusion; Kusner DJ et al.; Advances in therapy for tuberculosis will require greater understanding of the molecular mechanisms of pathogenesis and the human immune response in this disease . Exposure of Mycobacterium tuberculosis-infected human macrophages to extracellular ATP (ATP(e)) results in bacterial killing, but the molecular mechanisms remain incompletely characterized . In this study, we demonstrate that ATP(e)-induced bactericidal activity toward virulent M . tuberculosis requires an increase in cytosolic Ca(2+) in infected macrophages . Based on our previous work with primary infection of human macrophages, we hypothesized that the Ca(2+) dependence of ATP-induced killing of intracellular M . tuberculosis was linked to promotion of phagosome-lysosome fusion . Using confocal laser-scanning microscopy, we demonstrate that ATP(e) induces fusion of the M . tuberculosis-containing phagosome with lysosomes, defined by accumulation of three lysosomal proteins and an acidophilic dye . Stimulation of phagosome-lysosome fusion by ATP(e) exhibited distinct requirements for both Ca(2+) and phospholipase D and was highly correlated with killing of intracellular bacilli . Thus, key signal transduction pathways are conserved between two distinct models of human macrophage antituberculous activity: primary infection of naive macrophages and physiologic stimulation of macrophages stably infected with M . tuberculosis.

Med Sci Monit, 2001 Sep-Oct, 7(5), 878 - 83
CD4/CD8 lymphocytes in BALF during the efferent phase of lung delayed-type hypersensitivity reaction induced by single antigen inhalation; Grubek-Jaworska H et al.; BACKGROUND: The precise mechanisms involved in the pathogenesis of hypersensitivity pneumonitis (HP) have not been identified . HP is characterized by inflammatory lymphocytic alveolitis and a remarkable increase in T-lymphocytes detected in bronchoalveolar lavage fluid (BALF) . It is suggested that both CD4+ and CD8+ T cells may contribute to the pathogenesis of HP . Experiments on animal models suggest that cell mediated immunity (CMI) is more important for the pathogenesis of HP than complex-mediated immunity, but the relationship between the subsets of BALF lymphocytes and humoral or cell-mediated allergic reactions is still not clear . The aim of our study was distinguish CD4+ and CD8+ T cells in BALF lymphocytes during a delayed-type hypersensitivity (DTH) reaction in the lung . MATERIAL AND METHODS: The experiment was performed on guinea pigs sensitized with BCG vaccine and subjected to a single inhalation of tubercle bacilli antigens (tuberculin) . 24 hours after tuberculin provocation (at the time of maximum lymphocyte infiltration), bronchoalveolar lavage was performed on both sensitized and non-sensitized (control) animals . The total cell count was estimated, and a differential microscopical examination of BAL-fluid cells was performed, along with the phenotyping of BALF lymphocytes (by flow cytometry) . RESULTS: In the BALF of the sensitized animals, as compared to the controls, there was a statistically significant increase in the percentage and absolute count of T-lymphocytes, CD4+ and CD8+ . The CD4 / CD8 ratio in both groups did not differ significantly and was individually variable (2.94I0.72 SEM in the experimental group, vs 4.41I1.29 SEM in the control group) . CONCLUSIONS: Both CD4+ and CD8+ lymphocytes (with some predominance of helper cells) participate in the efferent phase of the delayed type hypersensitivity reaction in the lung induced by antigen inhalation.

Ophthalmology, 2001 Sep, 108(9), 1580 - 5
Ocular manifestations of tuberculosis; Sheu SJ et al.; OBJECTIVE: To present the clinical and histopathologic findings in five cases of tuberculosis (TB) with various ocular manifestations . DESIGN: Observational case series . METHODS: Retrospective review of clinical findings, course, and treatment of five patients . Diagnostic techniques, including biomicroscopic, histopathologic, and molecular biologic test results, are presented . MAIN OUTCOME MEASURES: Visual acuity, slit-lamp biomicroscopy, indirect ophthalmoscopy, and fluorescein angiography results . RESULTS: The ocular manifestations of TB in our patients included panophthalmitis, endophthalmitis, posterior uveitis with choroidal tubercles, keratitis, and a lid mass . Mycobacterium tuberculosis was identified in four cases in ocular specimens using acid-fast bacilli microscopy and in three cases by culture . Rapid diagnosis using polymerase chain reaction was obtained in one case . Extraocular foci of TB were identified in three cases with an intraocular infection at presentation . No patients had the human immunodeficiency virus (HIV) and none were immunocompromised . Two eyes could not be saved using antituberculous treatment because of delayed diagnosis and treatment . CONCLUSIONS: In this age of the HIV pandemic, TB is becoming more common . Because it is curable, heightened awareness and better understanding of the disease's ocular manifestations should be of concern to all ophthalmologists.

Klin Lab Diagn, 2001 Jul, (7), 50 - 3
{Mycobacterium tuberculosis detection by polymerase chain reaction and identification of M . tuberculosis strain}; Beklemishev AB et al.; A rapid multiprimer PCR method for detection of Mycobacterium tuberculosis complex (MTC) and simultaneous identification of M . tuberculosis in clinical samples has been developed . The method is based on simultaneous amplification of two targets: a 401 bp region from the mtp40 species-specific gene sequence of M . tuberculosis and a 544 bp fragment from the RD1 genome region which is specific for MTC but absent in BCG strains . Polymerase inhibitors in this study were detected by internal control in each test . Detection sensitivity was 25 copies of M . tuberculosis genomic DNA . Seven methods for isolation of mycobacterial DNA were compared and the technique with chloroform extraction was selected as the most efficient . The proposed method was used for analysis of 37 clinical samples and the results were compared with the results of culturing, acid-fast bacilli staining, and clinical diagnosis . The method proved to be sufficiently sensitive and specific for detection of mycobacterial DNA . Moreover, in countries with only two main pathogenic species of MTC circulating (M . tuberculosis and M . bovis) this method can be used for differentiation of these two species.

Eur Respir J, 2001 Aug, 18(2), 376 - 80
The tuberculin skin test in reaction to immunological in vitro reactions in BCG-vaccinated healthcare workers; Fjallbrant H et al.; The aim was to study the tuberculin skin test in relation to immunological in vitro reactions in bacille Calmette-Guerin (BCG)-vaccinated healthcare workers . The present study was performed in Sweden, a country with a low incidence of tuberculosis, a high BCG vaccination efficacy and high tuberculin conversion rates . BCG-vaccinated healthcare workers (n=381) were tuberculin skin tested . From these, 11 subjects with negative tuberculin reactions (<6 mm) were matched for age and sex with 11 subjects with large positive reactions (> or = 15 mm) . Lymphocyte transformation and the production of interferon-gamma (IFN-gamma) were analysed after stimulation in vitro of peripheral blood mononuclear cells with tuberculin purified protein derivative, heat-killed tubercle bacilli and a culture filtrate from tubercle bacilli . In the tuberculin-positive group the lymphocyte transformation response was 2-3 times larger, and IFN-gamma production was 7-10 times larger, than in the tuberculin-negative group (p<0.001) . The present results suggest that a positive tuberculin skin test in bacille Calmette-Guerin-vaccinated subjects indicates a stronger immune response of the protective T-helper 1-type than does a negative test . In similar settings, the study supports the traditional practice of regarding the tuberculin skin test in bacille Calmette-Guerin-vaccinated subjects as an indicator of a protective immune response against tuberculosis.

Acta Leprol, 2000-2001, 12(1), 11 - 8
Histological changes in the nerve, skin and nasal mucosa of patients with primary neuritic leprosy; Suneetha S et al.; Primary neuritic leprosy (PNL) presents as a peripheral neuropathy with no visible skin patches and skin smears negative for acid fast bacilli . The pathogenesis of PNL is poorly understood . The aim of the study was to document the histological changes in the nerve, apparently normal skin and nasal mucosa in PNL and to study its significance to the pathogenesis of leprosy lesions . The study is based on a cohort of 208 PNL patients registered at the Schieffelin Leprosy Research and Training Centre, Karigiri . All patients had a nerve biopsy, 196 had a skin biopsy and 39 had a nasal mucosal biopsy . The findings reveal that PNL patients exhibit a spectrum of disease histologically in the nerve ranging from lepromatous to tuberculoid leprosy with a significant proportion (46%) manifesting a multibacillary leprosy histology . Findings in the apparently normal skin and nasal mucosa reveal that there are widespread changes due to leprosy in tissues such as the skin and nasal mucosa even when the disease appears clinically confined to a few nerves . PNL may be an early stage in the pathogenesis of the disease before the appearance of skin lesions . The number of nerves enlarged and lepromin status did not give any clue to the nature of underlying disease.

Nippon Ronen Igakkai Zasshi, 2001 Jul, 38(4), 481 - 3
{Oral care reduces pneumonia of elderly patients in nursing homes, irrespective of dentate or edentate status}; Yoshida M et al.; Aspiration of oral secretions and their bacteria, especially gram-negative bacilli, is increasingly being recognized as an important factor in pneumonia . We investigated whether oral care reduces the frequency of pneumonia in the institutionalized elderly . Nurses or caregivers cleaned patients' teeth with toothbrushes after each meal, and scrubbed the oropharynx with an applicator with povidone iodine (1%) every day . Dentists or dental hygienists gave professional care once a week . During follow up, pneumonia decreased significantly in patients with oral care . Oral care was of benefit for both edentate as well as dentate subjects, although periodontal diseases might cause pneumonia . We suggest that oral care may be useful in preventing pneumonia in institutionalized elderly, irrespective of dentate or edentate status.

Kekkaku, 2001 Jul, 76(7), 549 - 57
{Tuberculosis control strategy in the 21st century in Japan--for elimination of tuberculosis in Japan}; Aoki M; Modern tuberculosis control programme has been launched in 1951 by the major revision of the previous Tb . Control Law in Japan . Main control measures were BCG vaccination programme for tuberculin negatives, annual screening of Tb . by miniature radiophotography (MMR), charge free diagnosis and treatment of Tb . patients, registration and case-holding at Health Centres throughout the country and so on . Thanks to the efforts of the Government and people concerned, Tb . incidence has decreased with the annual reduction rate of 11% during 1961 and 1977 . However, Tb . decrease has stagnated after that, and it is increasing slowly in these 3 years since 1998 . Moreover, regional variations of Tb . incidence are considerable, and Tb . is concentrated in specific risk groups such as elderly persons, homeless, foreign born individuals and so on . However, the present Tb . Control measures were introduced prior to the discovery of most major anti-Tb . drugs and all modern internationally accepted Tb . Control strategies, so that it is strongly desired to improve the present control programme from rather classical present Tb . control measures to global standard one to overcome the resurgence of Tb . in Japan . At first, the author stressed that the priority of Tb . Control Programme should be changed according to the development of science and the change of epidemiological situations . BCG vaccination and Tb . screening by MMR might be very important when the annual risk of Tb . infection was very high--about 4% in 1950 . Now it is around 0.05% and the incidence of Tb . among 0-14 years of age is 1.1 per 100,000 so that the priority should be given on treatment of the detected cases instead of BCG vaccination or MMR . The doctors in the public health field should give more strong concern on clinical aspects of Tb . Control Programme at present . It was considered that the main urgent problems to be improved in the present Tb . control measures are as follows . 1 . It is strongly recommended to spread the global standard regimen with 2HRZE/4HR (E) more widely and rapidly . Because the standard regimen is used in only 50% of new smear positive cases at present although 15.3% of Tb . patients are 80 years or more, or 56.3% of them are 60 years or more, and the side effects by PZA are higher among elderly patients . 2 . Shortening of the hospitalization duration is required because 76.7% of newly detected bacilli positive cases are hospitalized at first, and the median of the period of hospitalization is 4 months, and 18.4% of them are hospitalized 6 months or more at present . 3 . DOT treatment has been introduced for special groups in the big cities in 2000 for the first time in Japan, but it is needed to spread DOT treatment more widely, for example, by increasing health insurance payment for the institutions where DOT treatment is being implemented . 4 . It is recommended to build special rooms to accept Tb . patient at general hospitals and/or university hospitals to avoid the neglect of Tb . by general medical doctors . 5 . Follow-up of Tb . patients after treatment completion at Health Centres is not needed now, because the relapse rate is so low . 6 . Indiscriminative screening programme for all the people aged 19 years old or more should be stopped, at least up to 39 years of age, because Tb . detection rate has become so low as 0.0069% at present . As Tb . decrease is so slow, or is increasing in some areas, that the contact surveys among the young aged 20 to 39 should be strengthened in the future . 7 . As Japan Anti-Tb . Association is being carrying out mass screening programme extensively at present, so that the Association has started to discuss the future health check system . Because of the rapid and constant increase of the lung cancer, the Association is discussing the method to detect the lung cancer, too . In any way, it is needed to focus the screening programme for special high risk groups instead of indiscriminative screening . 8 . BCG vaccination for infants should be continued a little more, because BCG vaccination can protect the development of 7 miliary Tb . and/or meningitis cases during 15 years if BCG is given 70% of the infants in 2000 . However, it was strongly recommended to stop re-vaccination of BCG, because it is not so effective, and disturb the diagnosis of Tb . infection by tuberculin testing . 9 . Treatment of latent Tb . infection will become more and more important, so that it's indication should be expanded to the adults in the future instead of the present indication up to 29 years of age . It is needed to revise Tuberculosis Control Low to improve control programme in Japan . The author hoped that the members of Japan Tuberculosis Society will promote the improvement and to support the Government to improve the Law.

Kekkaku, 2001 Jul, 76(7), 519 - 24
{The alternative or simultaneous excretion of different species of mycobacteria in atypical mycobacteriosis--15 cases in 7 years}; Hiraoka T et al.; Five hundreds and five cases of Mycobacterium tuberculosis infection admitted to our hospital during 7 years from 1993 to 1999 . Numbers of newly diagnosed cases were 390, of which 329 (84.4%) were bacilli positive with sputum examination on admission . The recurrent cases were 115, of which 55 (47.8%) were bacilli positive cases . During the same period, mycobacteria other than tuberculosis (atypical mycobacteria) were detected in the sputum of other 121 cases . The 63 (52.1%) out of these 121 cases were diagnosed as atypical mycobacteriosis (AM) due to repetitive detection of bacilli with fulfilled the criteria according to the Japanese Mycobacteriosis Research Group of the National Chest Hospitals . The ratio of 63 AM cases to 329 bacilli positive TB cases was 19.1%, but it range from 8.2% to 31.3% year by year . In these 63 AM cases, 9 (14.3%) cases excreted AM bacilli simultaneously or alternately with TB bacilli . The other 15 (23.8%) cases excreted different AM bacilli simultaneously or alternately . The changes or combinations with M . avium and M . intracellulare were most frequently observed (in 12 cases) . The manners of changes and combinations with different species were described in detail.

Scand J Infect Dis, 2001, 33(7), 483 - 7
Cytolytic T cells in the immune response to mycobacterium tuberculosis; Stenger S; Cytolytic T cells (CTL) are of paramount importance in immune defense against tumors and viruses . Work over the past decade has revealed that lysis of infected cells is also involved in protective immunity to bacteria and parasites, including Mycobacterium tuberculosis . Experiments involving gene-deleted mice and the characterization of CTL lines derived from tuberculosis patients suggest an important role of CTL in immunity to tuberculosis . More recently, the identification of an effector pathway of human CTL provided evidence for direct antimicrobial activity of CTL . This pathway involves the combined action of the pore-forming perforin and the antibacterial granulysin, both expressed in the granules of CTL . Granulysin binds to the bacterial cell surface, thereby disrupting the membrane and causing osmotic lysis . The relevance of this pathway for protection against intracellular pathogens is suggested by the expression of high amounts of granulysin in tissue from patients with tuberculoid leprosy, which are able to contain the spread of the bacilli . These findings support the current concept of designing novel vaccination strategies which elicit not only CD4 + T helper cells, but also CD8 + CTL with direct antibacterial activity.

J Bacteriol, 2001 Sep, 183(18), 5311 - 6
Microaerophilic induction of the alpha-crystallin chaperone protein homologue (hspX) mRNA of Mycobacterium tuberculosis; Desjardin LE et al.; Among the products that are expressed when Mycobacterium tuberculosis undergoes hypoxic shiftdown to nonreplicating persistence (NRP) is the alpha-crystallin chaperone protein homologue (Acr) . This expression coincides with the previously reported appearance of a respiratory type of nitrate reductase activity, the increase in glycine dehydrogenase activity, and the production of a unique antigen, URB-1 . In a timed sampling study, using a slowly stirred oxygen depletion culture model, we have demonstrated that the hspX mRNA that codes for Acr protein as well as the protein itself is induced just as the bacilli enter the microaerophilic NRP stage 1 (NRP-1) . In contrast to the induction observed for hspX mRNA, levels of 16S rRNA, fbpB mRNA (encoding the 85B alpha antigen), and aroB mRNA (encoding dehydroquinate synthase) demonstrate relatively small to no change upon entering NRP-1 . Acr protein was shown to be identical to URB-1 by Western analysis with anti-URB-1 antibody . The fact that antibody to Acr is found in a high percentage of tuberculosis patients suggests that the hypoxic shiftdown of tubercle bacilli to the NRP state that occurs in vitro, resulting in production of the alpha-crystallin protein, occurs in vivo as well . Simultaneous abrupt increases in hspX mRNA and Acr protein suggest that Acr protein expression is controlled at the level of transcription.

Probl Tuberk, 2001, (3), 22 - 9
{Caseous pneumonia: diagnosis, clinical aspects and treatment}; Mishin VIu; Fifty-two patients with caseous pneumonia (CP) (whose age ranged from 18 to 60 years) admitted to a specialized tuberculosis facility of the general health care system were followed up . On admission, all the patients had marked intoxication and bronchopulmonary manifestations of the disease . X-study revealed lesions to one lobe in 19.2%, to 2 lobes in 32.7%, to 3 lobes or more in 48.1% of the patients . At the same time, all the patients had destructive changes with large and giants caverns in 57.7% of cases . The sputa of all 52 patients exhibited Mycobacteria tuberculosis (MBT), whose primary drug resistance being established . The nonspecific microflora composed of gram-positive cocci, gram-negative bacilli, fungi of the genus Candida in over 60% of patients was found in the sputa in two thirds of cases . A retrospective analysis indicated that CP had been correctly diagnosed only in 11.5% of patients within the first fortnight, in 65.4% by the end of the first month and by the beginning of the second, and in 23.1% following 2 months . The main reason for untimely diagnosis of CP was no sputum test for MBT . An algorithm of early CP diagnosis at general hospitals was developed, which includes a compulsory test of 3 sputum smears by the Ziehl-Nielsen method in all patients with pneumonia on admission and 2 weeks after treatment with wide-spectrum antibiotics . Chemotherapy in CP patients was long-term and combined 4-5 essential drugs within the first 4 months with obligatory correction after there was evidence for drug resistance . A course of nonspecific antibiotic therapy and a developed complex of symptomatic and pathogenetic treatments were compulsory . By the cessation of bacterial isolation, the efficiency of combined treatment was 82.7%, but a question as to a surgical intervention that is a compulsory component of CP therapy always arose at the final stage.

Hernia, 2001 Jun, 5(2), 97 - 8
Bubo masquerading as an incarcerated inguinal hernia; Hodge KR et al.; A 44-year old, male, tattooed, leather jacket clad, Harley-Davidson motorcyclist arrived at the emergency room with a tender, irreducible mass, presenting at the external inguinal ring . In 1998, lung biopsy was read as miliary granuloma . No herniation was found on urgent preperitoneal exploration . Incision of the mass showed acid-fast bacilli . Culture later revealed Mycobacterium avium-intracellulare (MAI) . A blood count showed CD4 lymphopenia; HIV was negative and remains so 2 years later.

Folia Microbiol (Praha), 2001, 46(2), 165 - 71
Protein profile and biochemical properties of Bacillus circulans isolated from intestines of small free-living animals in Poland; Swiecicka I; Forty-seven strains of Bacillus circulans isolated from the intestinal tract of free-living small mammals from the Narvia and Biebrza National Park (NE Poland) were compared with the electrophoretic whole-cell protein patterns on the basis of SDS-PAGE and biochemical characteristic using API tests (50 CHB and 20E) . The strains were grouped into two clusters (I and II) at the similarity of protein pattern of 78% using the simple matching coefficient and clustering on unweighted pair group arithmetic average algorithm (SSM/UPGMA) . Each of the clusters comprised reference strains . The rods included in the cluster I are characterized by higher biochemical activity in comparison with the bacilli II.

Rev Esp Quimioter, 2000 Dec, 13(4), 366 - 73
{Cefepime in the treatment of osteomyelitis caused by Gram negative bacilli}; Barberan J et al.; We conducted a prospective, randomized, open-label trial to evaluate the efficacy and tolerability of cefepime in the treatment of osteomyelitis caused by Gram-negative bacilli . Hospitalized patients with diagnosis of osteomyelitis due to Gram-negative bacilli susceptible to cefepime were elegible for enrollment . Cefepime was administered intravenously or intramuscularly (2 g every 8 or 12 hours) . Microorganisms were considered susceptible to cefepime when the MIC was <8 mg/l . Forty-five patients with bone infections were enrolled, forty-three with osteomyelitis (22 chronic and 21 acute) and two with arthritis . In the per protocol analysis 42 patients were evaluated: 30 (71.4%) were cured . In the intent to treat analysis 45 patients were evaluated: 33 (73.3%) were cured . Our trial suggests that cefepime is as effective as other modern parenteral beta-lactam antibiotics in the treatment of osteomyelitis due to Gram-negative bacilli.

Lepr Rev, 2001 Jun, 72(2), 179 - 91
Response of Mycobacterium habana vaccine in patients with lepromatous leprosy and their household contacts . A pilot clinical study; Wakhlu A et al.; Single dose vaccination was carried out with Mycobacterium habana vaccine, 31 lepromatous leprosy cases receiving 1.5 mg (1.5 mg = 6.27 x 10(8) bacilli) and 36 household contacts randomly receiving 1.5, 2.0, 2.5 mg vaccine intradermally . Duration of study was 18 weeks . Vaccination induced lepromin conversion in 100% of lepromatous leprosy cases and lepromin negative household contacts and augmentation of lepromin reactivity in 100% of lepromin positive household contacts, which was stable for the 15 weeks duration of follow-up . The maximum augmentation in lepromin reactivity was obtained with 1.5 mg of vaccine, which is probably the supramaximal dose . Overall, post-vaccination, those without prior BCG vaccination scars showed higher mean values of lepromin augmentation . Local vaccination site changes included induration, ulceration, itching, pain and uncomplicated regional lymphadenopathy, all of which remitted spontaneously by 15 weeks . Systemic side-effects noted were pyrexia, ENL and jaundice, and were seen with no greater frequency than that reported in other vaccine trials . Overall, systemic side-effects were easily controlled and were not accompanied by clinically detectable nerve or ocular damage . The safety profile investigations revealed an increase in the mean values of Hb%, RBC count and PCV in household contacts and of PCV in lepromatous patients, post-vaccination . Alterations in the liver function tests were also observed in patients of lepromatous leprosy . Thus, M . habana vaccine appears to be useful in stimulating specific CMI against M . leprae as evidenced by increased lepromin reactivity.

Lepr Rev, 2001 Jun, 72(2), 171 - 8
Fine needle aspiration cytology (FNAC) of nerves in leprosy; Vijaikumar M et al.; Leprosy is primarily a disease of the peripheral nerves and a technique that is simpler than nerve biopsy is required to evaluate nerve involvement, especially in pure neuritic (PN) leprosy . This study was designed to evaluate the role of FNAC of the nerve in the diagnosis and classification of leprosy . A prospective study was carried out on 25 patients with clinically active leprosy and at least one thickened peripheral sensory nerve . Nerve aspirates were evaluated by May-Grunwald-Giemsa and Fite's staining . Lepromin test, slit skin smears (SSS), skin biopsies (except PN cases) and nerve biopsies were performed and compared with FNAC . FNAC of nerve from 23 cases (92%) yielded diagnostic aspirates . Acid fast bacilli were observed in six cases by FNAC . FNAC and nerve pathology were equally comparable with the other parameters evaluated . Based on the results, cytological criteria were developed for interpreting nerve aspirates and the cases were classified as paucibacillary (18), BB (2), BL (2), LL (1) and non-diagnostic (2) . All PN cases showed diagnostic paucibacillary type cytology . FNAC of the nerve yields diagnostic aspirates in leprosy comparable with nerve pathology and the proposed cytological criteria may be useful in classification of nerve aspirates.

Int J Tuberc Lung Dis, 2001 Aug, 5(8), 754 - 62
Diagnostic value of a nested polymerase chain reaction assay on peripheral blood mononuclear cells from patients with pulmonary and extra-pulmonary tuberculosis; Honore S et al.; SETTING: Saint Louis Hospital, Paris, France . OBJECTIVE: To determine the clinical relevance of detection of Mycobacterium tuberculosis DNA by nested polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMCs) in the rapid diagnosis of tuberculosis . DESIGN: Single-centre prospective case study of 90 hospitalised patients and 50 healthy subjects or blood donors from 1 January to 30 June 1998 . RESULTS: Twenty-three patients were diagnosed with tuberculosis (26.7%); 20 tuberculosis patients were culture-positive, with seven smear-positive for acid-fast bacilli . Sensitivity of smear, culture and nested PCR was 30.4 (7/23), 87 (20/23) and 30.4% (7/23), respectively . The specificity of smear and culture was 100%, and the specificity of the nested PCR was 96% in the healthy subjects . However, the specificity decreased to 83.6% in the hospitalised patients, with 11 nested PCR-positive patients without a diagnosis of tuberculosis . The sensitivity of the nested PCR was low in pulmonary tuberculosis (22.2%), but increased in pulmonary/extra-pulmonary tuberculosis (50%), extra-pulmonary tuberculosis (33%), and disseminated tuberculosis (33%) . CONCLUSION: The use of a nested PCR assay on PBMC may pose problems for the rapid diagnosis of tuberculosis with regard to low sensitivity and specificity . However, further studies are needed to confirm this technique as an alternative test for the diagnosis of paucibacillary forms of tuberculosis.

Kansenshogaku Zasshi, 2001 Jun, 75(6), 504 - 6
{A case of Mycobacterium avium lung infection with a pleural effusion}; Saitou M et al.; Pleural effusions seldom accompany nontuberculous mycobacterial infections . We reported one such case of M . avium lung infection with pleural effusion . A 40-year old male was admitted to our hospital complaining of right chest pain and general fatigue . His chest X-ray showed a consolidation in the right lower lung field . The day after admission, a right pleural effusion appeared . The fluid was exudative and microbiological examinations of the effusions, including staining and culturing, proved negative . However, one month afteradmission, acid fast bacilli were observed in his sputum and a subsequent sputum culture specimen revealed the presence of M . avium . Treatment with antimycobacterial agents was promptly commenced and the patient's effusion and lung consolidation was gradually resolved.

Kekkaku, 2001 Jun, 76(6), 485 - 9
{A case of Mycobacterium intracellulare lung disease occurred in a middle-aged man who complained hemoptysis with normal chest roentogenogram}; Sasaki S et al.; A 45-year-old man visited our clinic because of intermittent bloody sputum . The chest roentogenogram was normal, but the high-resolution computed tomography (HRCT) showed very small nodules and bronchiolitis adjacent to pleura in the upper right lung field . The bronchoscopic examination revealed blood-streaked bronchial secretion in the right upper lobe bronchus, and the cultures of the sputa and the bronchial washing specimen showed acid-fast bacilli identified as Mycobacterium intracellulare by DNA-DNA hybridization (DDH) method . This case was diagnosed as Mycobacterium intracellulare lung disease . The patient received isoniazid, levofloxacin, and clarithromycin for three years without clinical and bacteriological improvement . His hemoptysis and the number of colonies recovered from sputum cultures decreased without any medication later . The serial chest roentogenograms and HRCTs have showed no changes for 6 years after the diagnosis . This case may show some clues to elucidate the mechanism of the onset of Mycobacterium intracellulare lung disease without predisposing conditions.

Kekkaku, 2001 Jun, 76(6), 479 - 84
{A case of tuberculous peritonitis diagnosed by a direct smear of ascitic fluid complicated with an active pulmonary tuberculosis and intestinal tuberculosis}; Mizutani H et al.; We sometimes encounter difficulties in differentiating tuberculous peritonitis from other inflammatory disorders or ascites due to carcinomatous peritonitis . Acid-fast bacilli are very rarely detected in ascites . In this study, we reported a case of tuberculous peritonitis accompanied with active pulmonary tuberculosis in which acid-fast bacilli were detected in ascites . The patient was a 37-year-old single man who had been admitted to our hospital on February 28, 2000, because acid-fast bacilli were detected in sputum, faces and ascites by a direct smear . He had a lower abdominal distention and pain . His serum CA 125 level was high, 121 U/ml . Abdominal ultrasonography showed marked ascites in Douglas pouch . However adenosine deaminase level was not high in his ascites . During treatment by the combination chemotherapy with INH, RFP, EB, and PZA, serum CA 125 level was decreased.

Kekkaku, 2001 Jun, 76(6), 461 - 71
{A study on resistance of Mycobacterium tuberculosis to four first-line anti-tuberculosis drugs in Japan: comparison of results in the local facilities and in the reference laboratory, in 1997}; Hirano K et al.; Five years after the last survey of drug-resistant tuberculosis in Japan, a nationwide survey was conducted by the Tuberculosis Research Committee (Ryoken) . A total of 78 hospitals in various districts of Japan participated in this cooperative study . Each collaborating laboratory sent all the isolated mycobacterial cultures during June 1 to November 30, 1997 to the Research Institute of Tuberculosis (RIT), which is one of the Supranational Reference Laboratories of the WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance . At RIT identification and drug susceptibility of Mycobacterium tuberculosis isolates were reexamined . The RIT received a total of 2,167 cultures . Among them, 523 cultures were excluded from further examinations because of various reasons, such as growth of mycobacteria other than tubercle bacilli (MOTT, 453), mixed cultures of M . tuberculosis and MOTT (16), and contamination or non-viability (54) . Thus drug susceptibility test results were available for 1,644 cultures, including 47 from foreign-born people . In the local laboratories, the absolute concentration method using 1% Ogawa egg slant (standard method, 26 hospitals), its modified methods using a 48-well plate (Microtiter method, 29 hospitals) and a 16-well plate (Well-pack method, 7 hospitals), combination of above 2 or 3 methods (13 hospitals), and other method (3 hospitals) were used for drug susceptibility testing, and the proportion method using 1% Ogawa egg slant was used in the RIT . The results in the local laboratories were compared with those in the RIT . A high concordance rate (over 90%) was seen in the testing for 1 microgram/ml of isoniazid (INH), rifampin (RFP) and streptomycin (SM), but the rate was lower (under 90%) in the testing for 0.1 microgram/ml of INH and ethambutol (EMB) . However, there was no significant difference in the concordance rates according to the test drugs among methods for drug susceptibility testing used in the local laboratories . Median concordance rates between the results with the standard method, Microtiter method and Well-pack method in the local laboratories, and those in the RIT were 95.9%, 93.2% and 96.4% respectively . Relatively lower concordance rates were seen in the laboratories using the Microtiter method related to high overestimation rates (median overestimation rate of 5.3%), compared with 1.2% and 2.3% in the laboratories using the standard method and Well-pack method, respectively . However, relatively lower concordance rates (less than 90%) were seen in the laboratories using any of the three methods, indicating that there are variations among facilities . Part of the results concerning the resistance patterns to four first-line anti-tuberculosis drugs were reported elsewhere.

Kekkaku, 2001 Jun, 76(6), 455 - 60
{A study on family infection of tuberculosis}; Hamajima I; The guideline for contacts examination and chemoprophylaxis for tuberculosis was revised in 1993 . Secondary cases and chemoprophylaxis cases found by the contacts examination of family members were analyzed in Sapporo City . The average age of index cases and of secondary cases has become higher . Many secondary cases were found among spouse of the index case and children of male index case . Chemoprophylaxis cases were found mostly among children of male or female index cases and grandchildren of male index cases . The results of family contacts examination carried out during the period from 1994 to 1999 were compared with those from 1987 to 1992 . The incidence of family infection of tuberculosis and the number of newly infected persons and of secondary cases have decreased in the latter period . The time interval from the registration of the index case to the detection of secondary cases has become shorter, and the ratio of sputum smear positive cases among secondary cases has also decreased . Male to female ratio of the index cases was 2:1, that of secondary cases was 3:5, and that of children indicated chemoprophylaxis was 4:5 . All secondary cases found by the contacts examination carried out within 6 months after the previous examination were sputum smear negative for tubercle bacilli, and there were a few smear positive cases among secondary cases who were found by the examination with longer interval with the previous examination . Such cases with longer interval from the last periodic examination was found often among younger cases below 40 years of age . Most smear negative tuberculosis cases were found by the family contacts examination, while most smear positive cases were found by symptomatic visit to doctors.

Microbios, 2001, 106(413), 55 - 67
Mycolic acids from Mycobacterium tuberculosis: purification by countercurrent distribution and T-cell stimulation; Goodrum MA et al.; Bacterial cell wall lipids are recognized as immunostimulatory molecules which make an important component of vaccines against bacterial diseases . Even mycolic acids, forming the waxy outer layer of the bacilli which cause tuberculosis, have been shown to stimulate human CD4/8 double negative T-cells . The role of these cells in resistance to tuberculosis is currently still debated . In this work, a method is described to purify mycolic acids from bacterial crude extracts in a single step using countercurrent distribution . Mycolic acids obtained in this way approach 100% purity and stimulate both double negative and CD4 positive T-cells in peripheral blood leucocytes obtained from healthy human donors . Stimulation of CD4 cells by mycolic acid antigens has not been reported before, emphasizing the potential importance of mycolic acids in the context of the fight against tuberculosis.

Am J Clin Pathol, 2001 Aug, 116(2), 225 - 32
Clinical and pathologic features of Mycobacterium fortuitum infections . An emerging pathogen in patients with AIDS; Smith MB et al.; The clinical and pathologic features of Mycobacterium fortuitum infection in 11 patients with AIDS were characterized . Nine patients had cervical lymphadenitis; 2 had disseminated infection . The infection occurred late in the course of AIDS, and the only laboratory abnormality seen in more than half of patients (7/11) was relative monocytosis . Absolute monocytosis also was seen in 4 of 11 patients . In both cytologic and histologic preparations, the inflammatory pattern was suppurative with necrosis or a mixed suppurative-granulomatous reaction . M fortuitum, a thin, branching bacillus, stained inconsistently in direct smear and histologic preparations . Staining was variable with Gram, auramine, Brown-Hopps, Gram-Weigert, Kinyoun, Ziehl-Neelsen, modified Kinyoun, and Fite stains . Organisms, when present, were always seen in areas of suppurative inflammation . Incorrect presumptive diagnosis, based on misinterpretation of clinical signs and symptoms or on erroneous identification of M fortuitum bacilli as Nocardia species, led to a delay in proper therapy for 7 of 11 patients . Definitive therapy after culture identification resulted in complete resolution of infection in all patients except 1.

J Vet Diagn Invest, 2001 May, 13(3), 255 - 8
Septicemia associated with Stenotrophomonas maltophilia in a West African dwarf crocodile (Osteolaemus tetraspis subsp . tetraspis); Harris NB et al.; A 17-year-old male captive West African dwarf crocodile (Osteolaemus tetraspis subsp . tetraspis) died 1 month after fighting with a penmate . Abrasions were present on the head and mandible . Necropsy revealed a vegetative valvular lesion of the left atrioventricular valve, miliary foci of necrosis in the endocardium and myocardium, multiple duodenal and rectal ulcers, and serous atrophy of body fat . Stenotrophomonas maltophilia was isolated in pure culture from lung, liver, and kidney . Gram-negative bacilli were seen histologically in the valvular lesion and in foci of necrosis in the myocardium, liver, spleen, pancreas, kidney, and intestine . Septic thrombi in multiple tissues, arteritis, and pneumonia were additional histologic lesions . Findings indicated that the crocodile died from acute S . maltophilia septicemia, although the primary site of infection was not determined . Stenotrophomonas maltophilia is ubiquitous in the environment and is recognized as an important nosocomial pathogen in humans.

Int J Lepr Other Mycobact Dis, 2001 Mar, 69(1), 21 - 5
PCR-restriction fragment length polymorphism analysis (PRA) of Mycobacterium leprae from human lepromas and from a natural case of an armadillo of Corrientes, Argentina; Zumarraga MJ et al.; Polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis (PRA) which relies on the amplification of a 439-bp portion of the hsp65 gene present in all mycobacteria, followed by two distinct digestions (with BstEII and HaeIII) of the PCR product, offers a rapid and easy alternative that allows identification of the species without the need for specialized equipment . Wild leprosy in the nine-banded armadillo (Dasypus novemcinctus) is characterized by the presence of multiple bacilli in internal organs such as lymph nodes, spleen and liver, as well as in nerves and skin . We could observe this in 9 out of 132 animals captured in Corrientes, Argentina, an area endemic for leprosy in humans . Mycobacterium leprae were recognized in those naturally infected animals through different techniques . Three samples of extracted DNA of the mycobacteria present in the spleen, liver and popliteal lymph node of a naturally infected animal during the Experimental Program in Armadillo (PEA) and three samples of human lepromas were processed by PRA . The patterns of the six samples analyzed were identical and were characteristic of M . leprae . These studies, made for the first time in Argentina, corroborate the initial discoveries in South America made by our investigative group on the detection of armadillos naturally infected with the Hansen bacillus.

Int J Lepr Other Mycobact Dis, 2001 Mar, 69(1), 1 - 12
Viable M . leprae as a research reagent; Truman RW et al.; Mycobacterium leprae remain a rare research resource . They cannot be cultivated on artificial media, and the only established means to quantify viability of M . leprae has been by its relative growth in the foot pads of conventional mice (MFP) . The MFP method is technically difficult and requires several months to yield results . More effective methods are needed . We examined the association between M . leprae's ability to oxidize 14C-palmitate in axenic culture and the MFP growth results of a large number of suspensions . Oxidative activity was assessed by radiorespirometry (RR) using the Buddemeyer-type biphasic culture vessels containing 7H12 liquid medium and 14C-palmitate, or with commercially prepared BACTEC 12B vessels containing the same medium . The RR results were highly correlated (r = 0.71) with the growth level that each M . leprae suspension achieved by the MFP technique . In using this technique to examine the effects that many common laboratory practices have on M . leprae viability, we found that viability varies markedly between bacillary suspensions derived from different hosts and tissues . The highest viabilities were obtained with bacilli from moderately enlarged nude MFP (< 1 g) . Viability tended to be lower among very large nude MFP or long-duration infections and from armadillo tissues . After their harvest from host tissues, leprosy bacilli lost viability quickly . Suspensions stored in 7H12 liquid medium retained < 1% of their viability within 3 weeks of harvest, and freezing bacillary preparations or incubating them at 37 degrees C resulted in nearly an immediate equivalent loss in metabolic activity and viability . M . leprae viability is maintained best when bacilli are stored for only short periods of time at 4 degrees C-33 degrees C . Palmitate oxidation is a rapid, reliable and objective means by which to estimate the viability of M . leprae and can be used effectively as a surrogate for the conventional MFP technique in many studies.

J Med Microbiol, 2001 Aug, 50(8), 675 - 81
Identification of catalase-like activity from Mycobacterium leprae and the relationship between catalase and isonicotinic acid hydrazide (INH); Kang TJ et al.; As Mycobacterium leprae proliferate inside macrophages, it has been speculated that catalase encoded by katG may protect the bacilli from deleterious effects of peroxide generated from the macrophage and may also play a crucial role in the survival of M . leprae in vivo . However, unlike that of M . tuberculosis, the katG of M . leprae has been reported to be a pseudogene, implicating that isoniazid, which is activated to a potent tuberculocidal agent by catalase, is unlikely to be of therapeutic benefit to leprosy patients . These results raise a question as to how M . leprae avoids H202-mediated killing inside macrophages . To understand the survival of M . leprae in macrophages, the present study attempted to detect catalase-like activity in M . leprae . Catalase-like activity was found in M . leprae cell lysate by the diaminobenzidine (DAB) staining method with non-denaturing polyacrylamide gel electrophoresis . An ammonium sulphate precipitation study revealed that the catalase-like activity was precipitable with 80% ammonium sulphate . The effect of isoniazid (INH) on M . leprae growth was also tested by RT-PCR and radiorespirometric assay to examine catalase-like activity in M . leprae, because INH was activated by catalase . It was found that the viability of M . leprae was decreased at a concentration of 20 microg/ml by radiorespirometric assay and it was inhibited at higher concentrations as determined by RT-PCR . These data suggest that a catalase-like activity other than that encoded by katG is present in M . leprae.

Zhonghua Jie He He Hu Xi Za Zhi, 1998 Oct, 21(10), 588 - 9
{Distribution and drug-resistance of 345 strains gram-negative bacilli from hospitalized patients with lower respiratory tract infections diseases}; Li Y et al.; OBJECTIVE: To investigate the distribution and drug-resistance of gram-negative bacilli in respiratory ward . METHOD: (1) Drug-resistance was tested by Kirby-Bauer disk sensitivity method and MIC by agar doubling dilution . (2) beta-lactanase was tested by nitrocephin . (3) ESBLs was tested by E test . RESULT: (1) Among 345 strains gram-negative bacillus, klebsiella pneumoniae and pseudomonas aeruginoea was about 39.7%, 17.4%, respectively . (2) Drug-resistance rate of cefotaxime dropped from 42.9% (6 years ago) to 8.2% . (3) beta-lactamase positive rate of 345 strains bacilli was 54.5% . (4) ESBLs was negative . CONCLUSION: (1) Klebsiella pneumoniae and pseudomonas aerugries were the main pathogens in lower respiratory tract infection in hospital . (2) Although we didn't find the evidence of ESBLs, we should pay more attention to it in the future.

J Clin Pathol, 2001 Aug, 54(8), 613 - 6
Identifying sputum specimens of high priority for examination by enhanced mycobacterial detection, identification, and susceptibility systems (EMDISS) to promote the rapid diagnosis of infectious pulmonary tuberculosis; Freeman R et al.; AIMS: To compare clinical information and sputum microscopy as methods for the selection of samples for enhanced mycobacterial detection, identification, and susceptibility systems (EMDISS) to promote the rapid diagnosis of patients with infectious pulmonary tuberculosis . METHODS: Two thousand, two hundred and sixty four specimen request forms were examined for clinical details, which were then used to identify specimens likely to yield Mycobacterium tuberculosis on culture . These results were compared with the results of sputum microscopy for acid fast bacilli (AFB) . Both methods were assessed against the results of culture using a combination of continuous automated mycobacterial liquid culture (CAMLiC) and conventional solid culture . RESULTS: Classification based on clinical details was an inefficient method of identifying high priority specimens for EMDISS . Although, when given, clinical details were often consistent, a substantial proportion of specimens arrived with no details . This approach would result in the referral of at least 16% of the workload but lead to the detection by culture of only 46% of the M tuberculosis present within it . In contrast, microscopy for AFB defined a much smaller number of specimens (4.8% of the total), which contained 90% of the M tuberculosis isolates . CONCLUSIONS: Microscopy for AFB is the most efficient method for defining sputum specimens suitable for referral for enhanced mycobacteriological techniques . However, it is essential that the methods used for smear preparation and microscopy are of the highest possible standard, otherwise some patients with infectious pulmonary tuberculosis will be denied, unnecessarily, the benefits of important advances in mycobacteriology.

J Clin Microbiol, 2001 Aug, 39(8), 2864 - 72
Characterization of clinical isolates of Klebsiella pneumoniae from 19 laboratories using the National Committee for Clinical Laboratory Standards extended-spectrum beta-lactamase detection methods; Steward CD et al.; Extended-spectrum beta-lactamases (ESBLs) are enzymes found in gram-negative bacilli that mediate resistance to extended-spectrum cephalosporins and aztreonam . In 1999, the National Committee for Clinical Laboratory Standards (NCCLS) published methods for screening and confirming the presence of ESBLs in Klebsiella pneumoniae, Klebsiella oxytoca, and Escherichia coli . To evaluate the confirmation protocol, we tested 139 isolates of K . pneumoniae that were sent to Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology) from 19 hospitals in 11 U.S . states . Each isolate met the NCCLS screening criteria for potential ESBL producers (ceftazidime {CAZ} or cefotaxime {CTX} MICs were > or =2 microg/ml for all isolates) . Initially, 117 (84%) isolates demonstrated a clavulanic acid (CA) effect by disk diffusion (i.e., an increase in CAZ or CTX zone diameters of > or =5 mm in the presence of CA), and 114 (82%) demonstrated a CA effect by broth microdilution (reduction of CAZ or CTX MICs by > or =3 dilutions) . For five isolates, a CA effect could not be determined initially by broth microdilution because of off-scale CAZ results . However, a CA effect was observed in two of these isolates by testing cefepime and cefepime plus CA . The cefoxitin MICs for 23 isolates that failed to show a CA effect by broth microdilution were > or =32 microg/ml, suggesting either the presence of an AmpC-type beta-lactamase or porin changes that could mask a CA effect . By isoelectric focusing (IEF), 7 of the 23 isolates contained a beta-lactamase with a pI of > or =8.3 suggestive of an AmpC-type beta-lactamase; 6 of the 7 isolates were shown by PCR to contain both ampC-type and bla(OXA) genes . The IEF profiles of the remaining 16 isolates showed a variety of beta-lactamase bands, all of which had pIs of < or =7.5 . All 16 isolates were negative by PCR with multiple primer sets for ampC-type, bla(OXA), and bla(CTX-M) genes . In summary, 83.5% of the K . pneumoniae isolates that were identified initially as presumptive ESBL producers were positive for a CA effect, while 5.0% contained beta-lactamases that likely masked the CA effect . The remaining 11.5% of the isolates studied contained beta-lactamases that did not demonstrate a CA effect . An algorithm based on phenotypic analyses is suggested for evaluation of such isolates.

J Vet Med B Infect Dis Vet Public Health, 2001 Jun, 48(5), 357 - 65
Fatal gastrointestinal infection with 'Flexispira rappini'-like organisms in a cat; Kipar A et al.; A 4-month-old male British Blue cat with catarrhal to haemorrhagic enteritis showed massive colonization of the stomach, small intestine and caecum with spiral-shaped bacilli . In the stomach, organisms were located in foveolae and gland lumina and within unaltered and degenerate epithelial cells . Inflammatory infiltration was moderate and T cell dominated . In the intestine, bacilli were found in the gut lumen, berween villi, in crypt lumina and within epithelial cells . Degeneration of crypt epithelial cells as well as crypt dilation and moderate to massive macrophage-dominated infiltration of the mucosa and submucosa were observed . Immunohistochemically, bacilli were positive with an antibody against Helicobacter . Ultrastructurally, the organisms strongly resembled 'Flexispira rappini', a spiral-shaped Helicobacter species known as a normal intestinal colonizer in dogs and mice.

Nefrologia, 2001 May-Jun, 21(3), 314 - 8
{Ileocecal tuberculosis during hemodialysis simulating carcinoma of the colon}; Garcia Marcos S et al.; Extrapulmonary tuberculosis is more frequent in hemodialysis patients than in the general population but intestinal localization is an unusual presentation of this infectious disease . We report a 60 year old patient on regular hemodialysis with intestinal tuberculosis masquerading as colon cancer . The patient presented with rectal bleeding, abdominal pain and fever and the radiological findings were compatible with ileocecal carcinoma . After surgery histological examination showed non-caseating granulomas but mycobacterial culture was not available . We performed a colonoscopy and obtained a biopsy of colonic mucosa for culture and other analyses . We identified acid-fast bacilli with Ziehl-Neelsen staining of formaldehyde preserved, paraffin-embedded tissue from the hemicolectomy and the colonic mucosal biopsy . Treatment with isoniazid, rifampicin and pyrazinamide for nine months was successful and well tolerated . Intestinal tuberculosis is a rare entity that we must keep in mind in a patient with abdominal pain, unexplained fever, digestive bleeding and particularly with a positive tuberculin reaction . When culture is not possible we can obtain intestinal samples by colonoscopy and use appropriate staining of paraffin-embedded tissues.

Rev Esp Enferm Dig, 2001 Feb, 93(2), 77 - 86
Surgical microbiology in risk patients undergoing abdominal surgery; Iturburu IM et al.; INTRODUCTION: Incidence of surgical infection is greater among elderly patients undergoing surgery than among the general population, with a serious compromise regarding morbidity and mortality in this group of patients with increased risk . AIM OF THE STUDY: To determine the microbiological features of surgical infections in patients over 65 years of age compared with those younger than 65 . PATIENTS AND METHODS: Over the past 2 years, 2,064 patients underwent surgery in our Department . One thousand three hundred sixty seven of those patients (66.7%) had 65 years of age or less and 688 (33.3%) were over 65 . Patient characteristics regarding the type of surgery (degree of contamination) and the nature and type of the condition requiring surgery are reported . One hundred five hundred sixty two samples were submitted for microbiological study . The microbiological infectious pathogens and their characteristics were determined . The Chi-square test was used for the analysis of potential differences related to the age of the patient . RESULTS: The rate of samples sent for microbiological study is higher among patients over 65 . In addition, these patients show both quantitative and qualitative differences in their infectious microbiological spectrum, mainly in clean-contaminated, contaminated and dirty surgical procedures . Fungal infections play a significant role in this group of patients . CONCLUSIONS: Facultative gram-negative bacilli, aerobic gram-positive cocci, and fungi are the main pathogens responsible of surgical infections in elderly patients, compared to all other patients, thus requiring specific antibiotic prophylactic and therapeutic regimes.

Rev Assoc Med Bras, 2001 Apr-Jun, 47(2), 125 - 8
{Isolated ileocecal tuberculosis simulating malignant neoplasia and Crohn's disease}; Bromberg SH et al.; BACKGROUND: Isolated ileocecal involvement by tuberculosis in the absence of pulmonar disease is rare in Brasil, therefore causes a diagnostic dilemma as it mimics colonic malignancy and Crohn's disease . METHODS: Between 1969 and 1989, eight patients with isolated hypertrophic ileocecal tuberculosis were treated by surgery in the Gastroenterology Surgery Department of the HSPE-FMO . The most common complaint among them was abdominal pain (100%) with associated symptoms of weight loss (62.5%); nausea, vomiting, fever and general weekness appeared in half of the patients . A right iliac fossa mass was present in seven (87.5% ) of them . The mean duration of symptoms was 14.7 month (range 5-36 months) . In all eight patients chest x-rays were negative for tuberculosis . Barium contrast studies showed abnormalities in all cases, but these could not be distinguished from carcinoma . Colonoscopy was helpful in establishing the diagnosis of suboclusive lesions of the ileocecal regions in three patients . Tuberculosis diagnosis was suspected in two of them because ofr the presence of granulomas in colonic biopsy material . RESULTS: Six patients were submitted to elective right hemicolectomy . The two remaining with suspect of tuberculosis were operated with signals of intestinal occlusion, and underwent a limited ileocaecal resection.The positive diagnosis of intestinal tuberculosis was made in all the patients by identification of acid-fast bacilli and by the presence of caseating granulomas in intestinal or lymph node tissue on histological examination . The outcome in all of them was favorable . They received treatment with three antituberculosis drugs over a twelve month period . CONCLUSION: Hypertrophic ileocecal tuberculosis must still be considered in the differential diagnosis of abdominal pathology localized in the right lower quadrant.

Tuberculosis (Edinb), 2001, 81(3), 229 - 42
Progressive pulmonary tuberculosis is not due to increasing numbers of viable bacilli in rabbits, mice and guinea pigs, but is due to a continuous host response to mycobacterial products; Dannenberg AM Jr et al.; Tuberculosis (TB) kills more people in the world today than any other infectious disease . A better vaccine to prevent clinical tuberculosis is greatly needed . Candidate vaccines are often evaluated by infecting rabbits, mice and guinea pigs by an aerosol of virulent tubercle bacilli and culturing their lungs for viable bacilli at various times thereafter . In all three species, however, the number of viable bacilli usually does not continuously increase until the host succumbs . The number of viable bacilli increases logarithmically for only about 3 weeks . Then, the host develops delayed-type hypersensitivity (DTH) and cell-mediated immunity (CMI), which keep the number of viable bacilli rather constant during the subsequent weeks . In the immunized host, DTH and CMI stop the logarithmic increase sooner than in the unimmunized controls, so that the stationary bacillary levels that follow are lower . This review analyzes host-parasite interactions in the lungs of rabbits, mice and guinea pigs . All three species cannot prevent inhaled fully virulent tubercle bacilli from establishing an infection, but they differ markedly in the type of the disease produced once it is established .

Respir Care, 2001 Aug, 46(8), 783 - 8
A comparison of two airway suctioning frequencies in mechanically ventilated, very-low-birthweight infants; Cordero L et al.; INTRODUCTION: Endotracheal suctioning of mechanically ventilated, very-low-birthweight infants for removal of respiratory secretions can be associated with morbidity . Routine endotracheal suctioning is inadvisable, but the safe minimum endotracheal suctioning frequency for prevention of airway obstruction has not been determined . HYPOTHESIS: Decreasing suctioning frequency from every 4 hours to every 8 hours (plus as needed) would have no clinically important effect on the primary outcomes (nosocomial bloodstream infection {BSI}, ventilator-associated pneumonia {VAP}, and bacterial airway colonization) or secondary outcomes (reintubation rates, need for postural drainage, severity of bronchopulmonary dysplasia {BPD}, neonatal mortality, duration of mechanical ventilation, and duration of hospitalization) . METHODS: We conducted a sequential retrospective study of 90 very-low-birthweight infants who were mechanically ventilated for longer than 7 days and who underwent endotracheal suctioning every 4 hours (plus as needed) and 90 similar infants who underwent endotracheal suctioning every 8 hours (plus as needed) . Two-pass endotracheal suctioning used during the study period required predetermined measurement of the suction catheter and prior instillation of saline . RESULTS: The 2 treatment groups were similar in demographic and clinical characteristics, including survival (90% and 85%), age at time of death (28 and 33 d), mean birthweight (926 and 934 g), gestational age (27 and 27 wk), duration of ventilation (29 and 27 d), and duration of stay (55 and 53 d) . Regardless of suctioning frequency, airway colonization with Gram-positive cocci occurred in the majority of patients by 2 weeks of life . Forty-three percent of the infants suctioned every 4 hours and 44% of those suctioned every 8 hours became airway colonized with Gram-negative bacilli . No Gram-negative bacilli species was more likely to be associated with either treatment group . VAP was diagnosed in 5 of the infants suctioned every 4 hours and in 9 of the infants suctioned every 8 hours . Nosocomial BSI occurred in 18 of the infants suctioned every 4 hours and in 21 of the infants suctioned every 8 hours . The difference in incidence of VAP and BSI was due to a epidemic that started before the change in suctioning frequency . During hospitalization, approximately one fourth of the patients in each group required postural drainage and were reintubated 11 and 10 times per 100 ventilator days, respectively . A comparable number of infants in each group developed severe BPD and were discharged home on oxygen . Suctionings per patient per ventilator day were 6 for the group suctioned every 4 hours and 4 for the group suctioned every 8 hours (p < 0.01) . CONCLUSIONS: A low-frequency suctioning regimen (every 8 hours plus as needed) can be implemented without increasing the incidence of nosocomial BSI, VAP, bacterial airway colonization, frequency of reintubation, need for postural drainage, severity of BPD, neonatal mortality, duration of mechanical ventilation, or duration of hospitalization . Although the minimum suctioning frequency for removal of unwanted respiratory secretions is yet to be established, a substantial reduction in endotracheal suctioning frequency appears to be safe.

Tuberculosis (Edinb), 2001, 81(1-2), 157 - 63
Genomics of Mycobacterium bovis; Gordon SV et al.; The imminent completion of the genome sequence of Mycobacterium bovis will reveal the genetic blueprint for this most successful pathogen . Comparative analysis with the genome sequences of M . tuberculosis and M . bovis BCG promises to expose the genetic basis for the phenotypic differences between the tubercle bacilli, offering unparalleled insight into the virulence factors of the M . tuberculosis complex . Initial analysis of the sequence data has already revealed a novel deletion from M . bovis, as well as identifying variation in members of the PPE family of proteins . As the study of bacterial pathogenicity enters the postgenomic phase, the genome sequence of M . bovis promises to serve as a cornerstone of mycobacterial genetics .

Zhonghua Yi Xue Za Zhi (Taipei), 2001 Mar, 64(3), 187 - 90
Multiple pleural nodules without effusion--a rare presentation of tuberculous pleurisy; Tang XN et al.; We report a rare case of tuberculous pleurisy presenting with multiple pleural nodules without associated effusion or parenchymal lung lesions . A 62-year-old man had multiple discrete pleural nodules in the right hemithorax on chest radiography without any clinical symptoms . Thoracoscopic biopsy of the pleural nodules revealed a caseous granuloma with acid-fast bacilli . The patient received antituberculous therapy, with resolution of tuberculomas on chest film within 2 months . To our knowledge, only two similar cases have been previously reported in the English literature, and our observation should lead to broadening of the spectrum of the differential diagnosis of multiple pleural nodules.

J Microbiol Immunol Infect, 2001 Jun, 34(2), 119 - 24
Infective endocarditis with neurologic complications: 10-year experience; Chen CH et al.; The impact of neurologic complications on clinical outcomes in infective endocarditis was assessed . Medical records of patients with infective endocarditis from January 1, 1987 through September 30,1998 were analyzed . Patients were divided into two groups: one with neurological complications and the other without . The outcomes of the two groups were compared using Fisher's exact test . Fifty-eight patients fulfilled the definite Duke criteria . There were 46 men and 12 women, ranging from 3 to 71 years of age with a mean of 40.6 years . Pathogens of infective endocarditis were documented by blood culture in 55 (94.8%) of 58 patients as follows: 52 with gram-positive cocci, two with gram-negative bacilli, and one with fungus . All 58 patients had initially received antimicrobial agents . Eight (13.8%) of the 58 patients had received surgical valvular replacement because of medical treatment failure . Overall, 16 (27.6%) of 58 patients died . Neurologic complications were either the chief complaint or one of the major presenting symptoms in 16 (27.6%) of the 58 patients . Patients with neurologic complications had a higher mortality rate (50% vs 20.9%, p = 0.025) than those without neurologic complications . The adjusted risk ratio for neurologic complications for a fatal event was 3.51 (95% CI = 1.1-11.18, p = 0.03) . Neurologic complications pose a significant problem in infective endocarditis . To reduce mortality, we recommend that more attention be paid to the treatment and prevention of the neurologic complications of infective endocarditis.

Indian Heart J, 2001 Jan-Feb, 53(1), 100 - 3
Fatal atypical mycobacterial infection in a cardiac transplant recipient; Ray R et al.; A 37-year-old female underwent heart transplantation for giant cell myocarditis . The patient died within three-and-a-half months of cardiac transplantation . Postmortem specimens from the heart and lung showed multiple necrotizing granulomas with numerous acid-fast bacilli . Polymerase chain reaction done on both the postmortem samples confirmed the presence of atypical mycobacterial infection . This fatal case of atypical mycobacteriosis in a cardiac transplant patient is reported for its rarity.

Radiographics, 2001 Jul-Aug, 21(4), 839 - 58; discussion 859-60
Thoracic sequelae and complications of tuberculosis; Kim HY et al.; Pulmonary tuberculosis is caused by Mycobacterium tuberculosis when droplet nuclei laden with bacilli are inhaled . In accordance with the virulence of the organism and the defenses of the host, tuberculosis can occur in the lungs and in extrapulmonary organs . A variety of sequelae and complications can occur in the pulmonary and extrapulmonary portions of the thorax in treated or untreated patients . These can be categorized as follows: (a) parenchymal lesions, which include tuberculoma, thin-walled cavity, cicatrization, end-stage lung destruction, aspergilloma, and bronchogenic carcinoma; (b) airway lesions, which include bronchiectasis, tracheobronchial stenosis, and broncholithiasis; (c) vascular lesions, which include pulmonary or bronchial arteritis and thrombosis, bronchial artery dilatation, and Rasmussen aneurysm; (d) mediastinal lesions, which include lymph node calcification and extranodal extension, esophagomediastinal or esophagobronchial fistula, constrictive pericarditis, and fibrosing mediastinitis; (e) pleural lesions, which include chronic empyema, fibrothorax, bronchopleural fistula, and pneumothorax; and (f) chest wall lesions, which include rib tuberculosis, tuberculous spondylitis, and malignancy associated with chronic empyema . These varieties of radiologic manifestations can mimic other disease entities . Therefore, recognition and understanding of the radiologic manifestations of the thoracic sequelae and complications of tuberculosis are important to facilitate diagnosis.

Keio J Med, 2001 Jun, 50(2), 91 - 9
Modern Japanese medical history and the European influence; Izumi Y et al.; Before the first European visited Japan in 1549, traditional Chinese medicine was mainly employed in Japan . Francisco de Xavier, a missionary of the Society of Jesus, tried to promote the introduction of Christianity by providing a medical service for Japanese citizens . However, Japan implemented a national isolation policy in 1639 and cut off diplomatic relations with the rest of the world, except Holland and China . For over 200 years, until the American admiral Matthew Perry forced Japan to open its doors in 1853, Japan learned about western medicine only from doctors of the Dutch merchants' office or from Dutch medical books . After 1853, Western medicine was rapidly introduced into Japan, and great achievements by Japanese medical doctors soon followed, such as the serum therapy for tetanus, the discovery of the plague and dysentery bacilli, the invention of Salvarsan for the treatment of syphilis, and the demonstration of the neurosyphilis spirochete.

Mol Gen Mikrobiol Virusol, 2001, (2), 13 - 9
{Regulation of extracellular phosphohydrolase biosynthesis in bacilli}; Morozova OV et al.; Under phosphate-deficient conditions, B . intermedius, B . pumilus, and B . thuringiensis secrete phosphohydrolases, including phosphomono-, phosphodiesterases, and guanyl-specific ribonucleases which cleave RNA molecules to nucleoside-3'-phosphatases . The enzymes are synthesized by phosphate-starved vegetative cells, which is not associated with sporulation . Using B . subtilis strains with mutation in the regulatory protein genes phoP and phoR, it was shown that these proteins regulate expression of B . intermedius, B . pumilus, and B . thuringiensis ribonuclease genes in B . subtilis cells . Genes of heterologous RNAses were activated in recombinant B . subtilis strains simultaneously with its own PHO regulon genes . Presumably a regulatory system homologous to B . subtilis two-component PhoP-PhoR signal transduction system functions in other representatives of the Bacillus genus.

Kekkaku, 2001 May, 76(5), 427 - 36
{Clinical study on the cases in which INH or RFP was discontinued during treatment for pulmonary tuberculosis}; Harada S et al.; Short course regimens; 2HRZ (E)(S)/4HR (E), 6HRS (E)/3-6HR and 6-9HR have been accepted as a standard chemotherapy (SC) for initial treatment of pulmonary tuberculosis in Japan . We studied the frequency of the treatment completion, the causes of the treatment failure and the outcome of the patients in whom INH or RFP was discontinued within 6 months after starting SC . The subjects included 597 newly diagnosed culture positive pulmonary tuberculosis patients admitted to 16 national hospital in 1996 . Results were as follows . 1 . In 47 (7.9%) of the 597 patients, either INH (19; 3.2%) or RFP (33; 5.5%) was discontinued . These 47 cases were defined as a SC incompleted group and the other 550 as a SC completed group . 2 . The patients in the SC incompleted group were seen more frequently in the ages of 20s (11.9%), 50s (10.9%), 60s (11.7%) or 70s (11.4%) . 21 (13.6%) of 154 female patients and 26 (5.9%) of 443 male patients were in the SC incompleted group . 3 . The causes of cessation of INH or RFP were drug side effects (33; 5.5%), drug resistance (10; 1.7%) and complications or underlying diseases (8; 1.3%) . 4 . Fever or eruption (19; 3.2%) and drug induced hepatitis (12; 2.0%) were frequently seen as drug related side effects causing the cessation of INH or RFP . 5 . The rate of culture negative conversion of TB bacilli at 6 months after the start of the treatment was 98.9% in the SC completed and 88.9% in the SC incompleted group respectively . In the SC incompleted group, there were three cases continuously positive and two other patients who relapsed and became culture positive again . In these five patients, INH or RFP was discontinued because of drug resistance.

Pneumologie, 2001 May, 55(5), 238 - 43
{Non-tubercular mycobacterial infection of the lungs due to Mycobacterium smegmatis}; Schreiber J et al.; Nontuberculous mycobacteriosis due to M . smegmatis is a rarity . We report on the case of a 51 year old male HIV-seronegative patient without predisposing bronchopulmonary disease, but with a state after gastrectomy and splenectomy who developed unproductive cough, night sweat and weight loss . The chest radiograph and thoracic CT showed wide-spread bilateral patchy infiltrations . Histological examination of transbronchial biopsies revealed chronic carnificating pneumonia . A perhoracic fine-needle biopsy showed caseating epitheloid cell granulomas with acid fast bacilli . These were identified as M . smegmatis by PCR with subsequent sequencing . Acid fast bacilli could not be detected microscopically neither in sputum nor in bronchial secretions, however M . smegmatis has been repeatedly detected by culture in these materials . In neither material tubercle bacilli have been detected by nucleic acid amplification (NAT) or culture . Immunologic investigations revealed a reduced number of CD4+ lymphocytes and a reduction of interferon alpha- and -gamma-synthesis by peripheral blood mononuclear cells . Treatment with Rifabutin, Ethambutol, Clarithromycin and Ofloxacin resulted in complete clinical and roentgenological resolution.

Diagn Microbiol Infect Dis, 2001 May-Jun, 40(1-2), 41 - 3
Activity of cefepime, cefotaxime, ceftazidime, and aztreonam against extended-spectrum-producing isolates of Klebsiella pneumoniae and Escherichia coli from Chilean hospitals; Zemelman C et al.; Resistance of Gram-negative bacilli to third-generation cephalosporins has been increasing due to the production of extended-spectrum beta-lactamases . In this work, the activities of cefepime, cefotaxime, ceftazidime, and aztreonam, alone and in association with clavulanic acid, against isolates of Klebsiella pneumoniae and Escherichia coli are compared . These isolates produce extended-spectrum beta-lactamases as shown by the synergy tests and by the decrease in the MICs of cephalosporins in the presence of clavulanic acid . Cefepime was the most active compound against these microorganisms . In addition, the microorganisms exhibited lower frequencies of resistance to this cephalosporin.

J Immunol, 2001 Jul 15, 167(2), 910 - 8
Toll-like receptor 2-dependent inhibition of macrophage class II MHC expression and antigen processing by 19-kDa lipoprotein of Mycobacterium tuberculosis; Noss EH et al.; Mycobacterium tuberculosis (MTB) induces vigorous immune responses, yet persists inside macrophages, evading host immunity . MTB bacilli or lysate was found to inhibit macrophage expression of class II MHC (MHC-II) molecules and MHC-II Ag processing . This report characterizes and identifies a specific component of MTB that mediates these inhibitory effects . The inhibitor was extracted from MTB lysate with Triton X-114, isolated by gel electroelution, and identified with Abs to be MTB 19-kDa lipoprotein . Electroelution- or immunoaffinity-purified MTB 19-kDa lipoprotein inhibited MHC-II expression and processing of both soluble Ags and Ag 85B from intact MTB bacilli . Inhibition of MHC-II Ag processing by either MTB bacilli or purified MTB 19-kDa lipoprotein was dependent on Toll-like receptor (TLR) 2 and independent of TLR 4 . Synthetic analogs of lipopeptides from Treponema pallidum also inhibited Ag processing . Despite the ability of MTB 19-kDa lipoprotein to activate microbicidal and innate immune functions early in infection, TLR 2-dependent inhibition of MHC-II expression and Ag processing by MTB 19-kDa lipoprotein during later phases of macrophage infection may prevent presentation of MTB Ags and decrease recognition by T cells . This mechanism may allow intracellular MTB to evade immune surveillance and maintain chronic infection.

Scand J Infect Dis, 2001, 33(5), 355 - 61
An evaluation of the diagnostic value of clinical and radiological manifestations in patients attending the addis ababa tuberculosis centre; Tessema TA et al.; Our investigation is based on 1,000 consecutive patients attending the routine outpatient services of the Addis Ababa Tuberculosis Demonstration and Training Centre (ATBDTC) in Addis Ababa during the period November 1996-March 1997 . By using sputum microscopy for acid-fast bacilli (AFB), chest radiography (CXR) and clinical assessment these patients had been diagnosed as having either AFB-positive pulmonary tuberculosis (PTB) (n = 139), AFB-negative PTB (n = 61) or non-tuberculosis (non-TB) (n = 800) . These three diagnostic groups were subsequently re-assessed by us with regard to selected demographic and clinical parameters, including CXR, in order to identify and weight markers significantly linked to proven PTB . The sum of the individual weights provided diagnostic scores (DS); the average DS for AFB-positive patients was 653.5 +/- 174 and that for non-TB patients was 219.1 +/- 138.7 . The calculated cut-off value between these two groups was 444 . Ten (7.2%) AFB-positive PTB patents had a DS below the calculated cut-off, while 46 (5.8%) of the 800 non-TB cases had diagnostic scores exceeding this value . Our DS system achieved 92.8% sensitivity and 94.3% specificity, with positive and negative predictive values of 73.7% and 98.7%, respectively . When plotting the individual DS values of the 61 AFB-negative TB patients, 24 (39.3%) fell below the cut-off . It is most likely that these patients did not have PTB . We conclude that there is a need for improved and standardized diagnostic approaches for TB-suspected patients who depend upon clinical and CXR criteria for correct diagnosis . Our score system may be helpful in this context.

Scand J Immunol, 2001 Jul-Aug, 54(1-2), 82 - 6
Liberation of soluble proteins from live and dead mycobacterial cells and the implications for pathogenicity of tubercle bacilli hypothesis; Wiker HG; Soluble proteins liberated from live M . tuberculosis are translocated through the cytoplasmic membrane to a 'periplasmic space' . For further export of proteins across the outer permeability barrier, it is necessary to postulate an excretion mechanism possibly involving some kind of porin . Observations of the repertoire of proteins in culture filtrates after liquid culture of M . tuberculosis show that a large repertoire of various kinds of proteins cross the outer permeability barrier of tubercle bacilli indicating that the excretion mechanism has a wide range of specificities for proteins . Culture filtrates of tubercle bacilli almost always contain both truly secreted proteins and cytoplasmatically-derived proteins . It is questionable whether cytoplasmic proteins can cross an intact cytoplasmic membrane . The simplest explanation for the appearance of cytoplasmic proteins in culture filtrates of tubercle bacilli would be that they are released after disintegration of the cytoplasmic membrane in dying or dead bacilli . Tubercle bacilli armed with secreted factors that may specifically inhibit innate and adaptive immune responses, excrete these from the periplasmic space of live bacilli . Unspecific in its character, the excretion mechanism also liberates proteins that are essential for building and maintaining the cell wall, thereby reducing the effectiveness of this process . This may be part of the explanation why M . tuberculosis and other pathogenic mycobacteria grow so slowly . Finally, it may be postulated that dormant or latent tubercle bacilli use their repertoire of secreted proteins to control their intracellular habitat and that bacterial cytoplasmic proteins would not be liberated from such bacilli . The consequence would be that only immune responses to secreted proteins would be effective for elimination of the dormant stage of infection . In a situation with active infection there will be considerable growth and turnover of bacilli with liberation of all kinds of immunogenic substances from the bacilli . In this situation immunity against cytoplasmic proteins would also be effective and immunity to cytoplasmic proteins should also be effective for control of the reactivation of latent disease because as soon as the bacilli start to grow there will also be a subpopulation of dead bacilli on the arena.

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