Air pollution and airborne infection with mycobacterial bioaerosols: a potential attribution of soot.

Atmospheric pollutants are hypothesized to enhance the viability of airborne microbes by preventing them from degradation processes, thereby enhancing their atmospheric survival. In this study, Mycobacterium smegmatis is used as a model airborne bacteria, and different amounts of soot particles are employed as model air pollutants. The toxic effects of soot on aerosolized M. smegmatis are first evaluated and excluded by introducing them separately into a chamber, being sampled on a filter, and then cultured and counted. Secondly, the bacteria-soot mixture is exposed to UV with different durations and then cultured for bacterial viability evaluations. The results show that under UV exposure, the survival rates of the low-, medium-, and high-soot groups are 1.1 (±0.8) %, 70.9 (±4.3) %, and 61.0 (±17.6) %, respectively. This evidence significantly enhanced survival rates by soot at all UV exposures, though the combinations of UV exposure and soot amounts revealed a changing pattern of survival rates. The possible influence by direct and indirect effects of UV-damaging mechanisms is proposed. This study indicates the soot-induced survival rate enhancements of M. smegmatis under UV stress conditions, representing the possible relations between air pollution and the extended pathogenic viability and, therefore, increased airborne infection probability.

Currents connecting communities: nearshore community similarity and ocean circulation.

Understanding the mechanisms that create spatial heterogeneity in species distributions is fundamental to ecology. For nearshore marine systems, most species have a pelagic larval stage where dispersal is strongly influenced by patterns of ocean circulation. Concomitantly, nearshore habitats and the local environment are also influenced by ocean circulation. Because of the shared dependence on the seascape, distinguishing the relative importance of the local environment from regional patterns of dispersal for community structure remains a challenge. Here, we quantify the "oceanographic distance" and "oceanographic asymmetry" between nearshore sites using ocean circulation modeling results. These novel metrics quantify spatial separation based on realistic patterns of ocean circulation, and we explore their explanatory power for intertidal and subtidal community similarity in the Southern California Bight. We find that these metrics show significant correspondence with patterns of community similarity and that their combined explanatory power exceeds that of the thermal structure of the domain. Our approach identifies the unique influence of ocean circulation on community structure and provides evidence for oceanographically mediated dispersal limitation in nearshore marine communities.

The stochastic nature of larval connectivity among nearshore marine populations.

Many nearshore fish and invertebrate populations are overexploited even when apparently coherent management structures are in place. One potential cause of mismanagement may be a poor understanding and accounting of stochasticity, particularly for stock recruitment. Many of the fishes and invertebrates that comprise nearshore fisheries are relatively sedentary as adults but have an obligate larval pelagic stage that is dispersed by ocean currents. Here, we demonstrate that larval connectivity is inherently an intermittent and heterogeneous process on annual time scales. This stochasticity arises from the advection of pelagic larvae by chaotic coastal circulations. This result departs from typical assumptions where larvae simply diffuse from one site to another or where complex connectivity patterns are created by transport within spatially complicated environments. We derive a statistical model for the expected variability in larval settlement patterns and demonstrate how larval connectivity varies as a function of different biological and physical processes. The stochastic nature of larval connectivity creates an unavoidable uncertainty in the assessment of fish recruitment and the resulting forecasts of sustainable yields.

Anti-tuberculosis drug susceptibility testing of Mycobacterium bovis BCG Tokyo strain.

SETTING: The Mycobacterium bovis bacille Calmette-Guérin (BCG) vaccine is the only vaccine against tuberculosis (TB), owing to its valuable protective effects and low virulence. However, it can occasionally cause systemic infection in immunocompromised hosts. Isoniazid (INH), rifampicin (RMP), streptomycin (SM) and ethambutol (EMB) are known to be effective anti-tuberculosis drugs and are used for the treatment of BCG infections. Unfortunately, there are few studies of the susceptibility of BCG vaccine strains to these drugs. OBJECTIVE: To measure the minimum inhibitory concentrations (MICs) of BCG Tokyo vaccine products for anti-tuberculosis drugs and assess vaccine safety in terms of drug susceptibility. DESIGN: We measured the MIC for one seed and five product lots of BCG Tokyo strain for INH, RMP, SM and EMB using Middlebrook 7H11 agar plates. RESULTS: The MIC results for INH were 0.06 and 0.125 mg/ml for the product and seed lots, respectively. The MIC results for RMP, SM and EMB were 0.25-0.5, 0.25 and 2-4 microg/ml, respectively. CONCLUSION: Our results indicate that the BCG Tokyo strain was susceptible to the major anti-tuberculosis drugs and treatable even in cases of severe adverse events, including systemic infection.

Tuberculosis and multidrug resistance in Zambian prisons, 2000-2001.

BACKGROUND: Data on prevalence of tuberculosis (TB) and multidrug-resistant tuberculosis (MDR-TB) in Zambian prisons are lacking. METHODS: Between January 2000 and July 2001, a case-finding study was performed in 13 Zambian prisons for pulmonary TB. Prisoners were administered a questionnaire to obtain demographic information. Information regarding housing density and diet was also collected. Three consecutive first morning sputum specimens were cultured for Mycobacterium tuberculosis. Antimicrobial resistance testing was performed by the resistance ratio method. RESULTS: A total of 1080 prisoners were recruited: 1055 were males and 25 females. Sputum from 245 (22.7%) prisoners yielded M. tuberculosis, including 168 (15.6%) with smear-positive disease. Based on a total prison population of 6118, the minimal prevalence of TB was 4.0%. There was a linear relationship between the proportion of prisoners evaluated and the prevalence of TB (R(2) = 0.9366) across facilities, suggesting that the true prevalence of TB may approach 15-20%. Resistance to at least one anti-tuberculosis drug was detected for 40 (23.8%) isolates, while MDR-TB was identified for 16 (9.5%) isolates. CONCLUSION: There is a high rate of pulmonary TB in Zambian prisons, with significant rates of drug resistance and MDR-TB, highlighting the need for active surveillance and treatment programs.

Nationwide survey on the prevalence of anti-tuberculosis drug resistance in the Republic of Yemen, 2004.

OBJECTIVES: To determine the prevalence of resistance to the four major anti-tuberculosis drugs, isoniazid, rifampicin, streptomycin and ethambutol, in Yemen. METHODS: Cluster sampling with probability proportionate to size was applied. Susceptibility to four major anti-tuberculosis drugs was examined. The proportion method using Löwenstein-Jensen medium or Ogawa medium was carried out. RESULTS: A total of 790 primary culture isolates from tuberculosis (TB) cases enrolled at the National Tuberculosis Institute, Yemen, were examined. In the confirmation culture at the supranational reference laboratory, 227 of them failed to grow on the secondary culture or were proved to be mycobacteria other than Mycobacterium tuberculosis and were excluded from further analysis. Among 563 cultures, 510 were obtained from new cases and 53 from previously treated cases. The prevalence of resistance to any four drugs was 9.8% (95%CI 7.0-12.5) among new cases and 17.4% (95%CI 12.0-33.5) among previously treated cases. The prevalence of multidrug-resistant TB was 3.0% (95%CI 1.5-4.5) among new cases and 9.4% (95%CI 0.2-18.7) among previously treated cases. CONCLUSION: The first nationwide prevalence survey on resistance to the four major anti-tuberculosis drugs in Yemen showed a relatively low prevalence of drug-resistant cases, but a high prevalence of multidrug resistance among new cases.

Preparation of mycobacteria-containing artificial sputum for TB panel testing and microscopy of sputum smears.

SETTING: Panel testing, blinded cross rechecking and on-site evaluation are the three methods for external quality assessment (EQA) of acid-fast bacilli (AFB) smear microscopy. Panel testing can provide data on laboratory capabilities prior to implementing a rechecking programme, assess the current status of laboratory performance and detect problems associated with diagnostic performance. Thus far, two methods for preparing panel test slides have been reported: these use real AFB-positive and -negative sputum treated with sodium hydroxide (NaOH) or N-acetyl-L-cysteine (NALC). OBJECTIVE: To evaluate the above methods and to develop a new method to prepare panel test slides with artificial sputum. DESIGN: Panel test slides were prepared using the NaOH and NALC methods. New artificial sputum preparation methods were developed and examined using a cultured monocyte cell line, cultured avirulent mycobacteria and methylcellulose or polyacrylamide gel as substrate. Smears prepared by the four methods were compared. RESULTS: Panel test slides prepared with NaOH and NALC methods are not macroscopically or microscopically similar to real smears. Our new artificial sputum is similar to real sputum in viscosity and macroscopic and microscopic appearance; it is also consistent in panel positivity grades. CONCLUSION: The artificial sputum described here could contribute to the EQA and training in tuberculosis laboratories or microscopy centres.

Early and delayed consequences of beta(2)-adrenergic receptor overexpression in mouse hearts: critical role for expression level.

BACKGROUND: Transgenic cardiac beta(2)-adrenergic receptor (AR) overexpression has resulted in enhanced signaling and cardiac function in mice, whereas relatively low levels of transgenically expressed G(alphas) or beta(1)AR have resulted in phenotypes of ventricular failure. Potential relationships between the levels of betaAR overexpression and biochemical, molecular, and physiological consequences have not been reported. METHODS AND RESULTS: We generated transgenic mice expressing beta(2)AR at 3690, 7120, 9670, and 23 300 fmol/mg in the heart, representing 60, 100, 150, and 350 times background betaAR expression. All lines showed enhanced basal adenylyl cyclase activation but a decrease in forskolin- and NaF-stimulated adenylyl cyclase activities. Mice of the highest-expressing line developed a rapidly progressive fibrotic dilated cardiomyopathy and died of heart failure at 25+/-1 weeks of age. The 60-fold line exhibited enhanced basal cardiac function without increased mortality when followed for 1 year, whereas 100-fold overexpressors developed a fibrotic cardiomyopathy and heart failure, with death occurring at 41+/-1 weeks of age. Adenylyl cyclase activation did not correlate with early or delayed decompensation. Propranolol administration reduced baseline +dP/dt(max) to nontransgenic levels in all beta(2)AR transgenics except the 350-fold overexpressors, indicating that spontaneous activation of beta(2)AR was present at this level of expression. CONCLUSIONS: These data demonstrate that the heart tolerates enhanced contractile function via 60-fold beta(2)AR overexpression without detriment for a period of >/=1 year and that higher levels of expression result in either aggressive or delayed cardiomyopathy. The consequences for enhanced betaAR function in the heart appear to be highly dependent on which signaling elements are increased and to what extent.

Comparative study on the use of solid media: Löwenstein-Jensen and Ogawa in the determination of anti-tuberculosis drug susceptibility.

A total of 53 Mycobacterium tuberculosis strains were examined to evaluate the consistency of susceptibility tests using two solid egg-based culture media, Ogawa and Löwenstein-Jensen (L-J), in order to assess the inter-media differences in susceptibility tests for anti-tuberculosis drugs. Fifty-three M. tuberculosis strains were tested for resistance against isoniazid, rifampicin, ethambutol, streptomycin and other alternative drugs on L-J and Ogawa media. Data from isoniazid (INH) revealed 31 strains were susceptible and 11 strains were resistant in L-J medium. On the other hand, one strain showed resistance against INH only on Ogawa medium. This gave a consistency ratio of 97.7% for INH between these two media. Similarly, the methods had 100% compatibility with rifampicin, 86.0% with ethambutol and 88.4% with streptomycin. When 41 of these strains were tested against ethionamide, enviomycin, para -aminosalicylate, sparfloxacin, and levofloxacin, the same numbers of strains were shown to be resistant or susceptible to L-J and Ogawa media. Similarly, the consistency ratio was 75.6% for kanamycin and 61.0% for capreomycin. There are inconsistencies between results obtained with L-J and Ogawa media, Ogawa medium had a tendency to produce results that indicated more resistant strains. However, the differences were statistically negligible.

Clinical evaluation of Amplicor Mycobacterium detection system for the diagnosis of pulmonary mycobacterial infection using sputum.

SETTING: The Amplicor Mycobacterium detection kit was evaluated for the diagnosis of active pulmonary mycobacterial infection using sputum. OBJECTIVE: To assess the clinical usefulness of the Amplicor Mycobacterium kit for the diagnosis of pulmonary tuberculosis and non-tuberculous mycobacterial infection in the country of medium prevalence. DESIGN: All the patients were diagnosed with bacterial, histopathological, and clinical 'gold standard'. The sensitivity and specificity for diagnosing clinically active pulmonary tuberculosis and Mycobacterium avium and Mycobacterium intracellulare infections were evaluated comparing Amplicor results and clinical diagnosis. RESULTS: A total of 1088 sputum specimens were collected from 780 in and out patients. Mycobacteria were recovered from 339 specimens by culture. The sensitivity and specificity of conventional culture method for the diagnosis of pulmonary tuberculosis were 60.2% and 99.8% respectively based on the number of patients. The figures for Amplicor were 61.8% and 97.4% respectively. There was no statistical significant difference between these methods. In rapidity, the Amplicor was significantly superior to the microscopy method in sensitivity. CONCLUSION: Patients with Amplicor positive and conventional negative result had mostly mycobacteria related diseases. The Amplicor positive result indicated mostly active mycobacterial infection and was clinically useful for rapid diagnosis.

Emergence of new HIV-1 subtypes other than Subtype C among antenatal women in Lusaka, Zambia.

Molecular epidemiology of HIV-1 in Zambia was investigated by direct sequencing of PCR products from samples collected from antenatal attendees in Lusaka, Zambia. One hundred and forty samples were initially screened for HIV, using antibody assays. Thirty-three (23.6%) samples were HIV-1 positive. Sequences of the HIV-1 env gp120 region were obtained from 28 of 33 (85%) HIV-1-positive samples. Twenty-six of the 28 sequences were HIV-1 env subtype C-like as previously reported. However, one HIV-1 env subtype D-like virus and one HIV-1 env subtype G-like virus were identified. This is the first time that these two HIV-1 env subtype viruses have been identified in Zambia, suggesting that more subtypes could be in existence.

Comparative study of amplicor Mycobacterium PCR and conventional methods for the diagnosis of pleuritis caused by mycobacterial infection.

OBJECTIVE: To assess the clinical significance of the Amplicor Mycobacterium system for the diagnosis of mycobacterial infection in patients with pleural fluid, and to compare its usefulness with that of conventional smear and culture methods. DESIGN: Pleural fluid specimens were obtained randomly from in-patients admitted to National Tokyo Hospital between January and December 1996. All the patients were diagnosed with bacterial, histopathological and clinical gold standard. The sensitivity and specificity for diagnosis of mycobacterial infection were evaluated. RESULTS: Seventy-five pleural fluid specimens were obtained. Conventional methods demonstrated a sensitivity of 30.6% and a specificity of 100%, while the Amplicor Mycobacterium demonstrated a sensitivity and specificity of 27.3% and 97.6% respectively. Lactic dehydrogenase, carcinoembryonic antigen, red blood cell, protein, glucose and types of inflammatory cells were not different in Amplicor positive and negative pleural fluid with mycobacterial infection. CONCLUSION: There was not much improvement in the accuracy of diagnosis when Amplicor Mycobacterium was used to diagnose mycobacterial pleuritis among various diseases with pleural fluid; however, the assay time was dramatically reduced with the use of Amplicor Mycobacterium.

Evaluation of PCR-based methods for the diagnosis of tuberculosis by identification of mycobacterial DNA in urine samples.

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

Potential use of Amplicor PCR kit in diagnosing pulmonary tuberculosis from gastric aspirate.

Polymerase chain reaction (PCR) detection of mycobacteria from gastric aspirate for the diagnosis of tuberculosis is not fully evaluated up to now. A total of 116 gastric aspirate specimens were collected from patients with suspected pulmonary tuberculosis. The breakdown of diagnosis was 67 pulmonary tuberculosis, 16 nontuberculous mycobacterial infection, 5 extra pulmonary tuberculosis, and 28 other lung diseases. The conventional methods were shown to have a sensitivity of 47.8% and a specificity of 79.6%; on the other hand, Amplicor had 34.9% and 97.0%, respectively. The Amplicor provided a more rapid and specific method for diagnosing tuberculosis and was more useful than the conventional.

Evidence of three clinical subgroups in patients with dual atrioventricular nodal pathways.

We attempted to test the hypothesis that dual atrioventricular (A-V) nodal pathways with second-degree atrioventricular block (2nd A-V block) present as a different clinical entity from those with A-V nodal reentranttachycardia (AVNRT). By evaluation with Holter monitoring (2.9 +/- 2.5 recordings/patient) and 12-lead electrocardiogram (11.9 +/- 11.6), 177 patients with dual A-V nodal pathways could be divided into three subgroups. Thirty-two patients had 2nd A-V block only (2nd A-V block group), 57 had AVNRT only (AVNRT group), 88 had neither 2nd A-V block nor AVNRT (silent group), and none had 2nd A-V block and AVNRT both. Electrophysiologic studies showed that the atrio-His interval was significantly greater (P < 0.0001) and the maximal 1:1 atrioventricular conduction rate was lower (P < 0.0001) in the 2nd A-V block group than in the other two groups. These differences were nullified after the administration of atropine. These results suggest that patients with dual A-V nodal pathways can be classified into three clinical subgroups based on the presence of either 2nd A-V block or AVNRT. We suggest also that patients of the 2nd A-V block group may have a more augmented vagal tone on the A-V node than the other two groups.

Amebiasis in Japanese homosexual men with human immunodeficiency virus infection.

We report cases of amebiasis in 6 human immunodeficiency virus (HIV)-positive male patients. Five were confirmed homosexuals while one was suspected. Three patients had liver abscess and 5 had colitis with duration of 10 days to months. The patients with liver abscess showed a lower incidence of abdominal pain but a higher incidence of concomitant diarrhea. Drainage therapy was effective for rapid afebrile results. Two invasive colitis cases died from perforation. This may have been due to delayed diagnosis. Invasive amebiasis is not common even in HIV-infected individuals. Among Japanese homosexual men, however, it may cause symptomatic diseases.

[A case report of miliary tuberculosis with tubercular meningitis diagnosed and followed by polymerase chain reaction method].

We report here a case of miliary tuberculosis with tubercular meningitis in which the polymerase chain reaction (PCR) amplification method was useful for its rapid diagnosis and follow up. A 70 year old male was hospitalized for further examination and treatment of diffuse small nodular shadows on the chest X-ray. After receiving antimicrobial therapy shadows still remained and he gradually lost visual acuity. He had no meningeal signs, and no remarkable finding on cranial CT. Cerebrospinal fluid examination showed increased cell number with predominantly lymphocytes. Cranial MRI (Gd DTPA) showed lateral ventricular ependymitis. Pulmonary tuberculosis and secondary tubercular meningitis were suspected, but we failed to detect microorganisms from the cerebrospinal fluid, gastric juice, sputum, and urine by the conventional method. However, by the polymerase chain reaction amplification method specific DNA fragments of Mycobacterium tuberculosis complex was detected from the cerebrospinal fluid, gastric juice, bronchoalveolar lavage fluid and serum. The final diagnosis of miliary tuberculosis with tubercular meningitis was established. We administered antitubercular drugs and observed the clinical course. He recovered and the polymerase chain reaction showed negative consequences in all samples. The judgement of PCR and the clinical course were compatible and parallel with the clinical course.

[A case of Enterococcus faecalis brain abscess in a patient with suprapharyngeal cancer].

A 55-year-old male was admitted to our hospital for the treatment of suspected brain abscess on March 18, 1989. The brain CT scan showed low density area with ring enhancement. After admission, both cefminox 2 g/day and clindamycin 500 mg/day were administered intravenously. On March 29, 1989, the patient developed cerebral herniation suddenly. Therefore emergency burr hole operation was done. Many gram-positive cocci were observed on gram-staining of the aspirated pus. Moreover neutrophils with phagocytosed gram-positive cocci were also observed. Culture of aspirated pus revealed colonies of gram-positive cocci which were subsequently identified as E. faecalis. Same species of bacteria were also isolated from fistula of right mastoid. From both findings of brain CT scan and data of MICs of E. faecalis, it may be suggested that direct infection developed from right mastoiditis to intracranial space. We experienced a severe case who developed cerebral herniation because of brain abscess due to E. faecalis which is an infrequent pathogen of brain abscess.

[A guide to prevent infectious disease due to MRSA].

Now in Japan, infectious diseases due to MRSA is an important problem to solve and it has a growing social interest. It is clearly associated with hospital infections. So most important thing for us is to prevent and treat properly hospital infections. As a result of thorough investigations in a hospital for aged persons and a general hospital, we developed some guidelines to prevent MRSA infections. It is necessary to have a prevention committee in the hospital which will do the followings: Reveal the present condition of hospital infections by thorough investigations, make appropriate choice of antibiotics, good care of patients having infectious diseases due to MRSA, good care of carriers, better use of disinfectants, and education of medical staffs.